JP2012131725A - Diphenyl sulfide derivative and medicine including the same as active component - Google Patents

Abstract

PROBLEM TO BE SOLVED: To provide a diphenyl sulfide derivative which is useful as a medicine having excellent S1P3 antagonist activity.SOLUTION: An investigation to create a compound having S1P3 antagonist activity has been zealously conducted, and as a result, it is found that a diphenyl sulfide derivative represented by general formula (1) [in the formula, Rrepresents a 1-6C alkyl group which is optionally replaced with 1 to 3 halogen atoms or a 1-6C lower alkoxy group, Rrepresents a 1-6C alkyl group or a 2-6C alkenyl group, Rrepresents a 1-6C acyl group or a carboxy group, X represents a methylene or an oxygen atom, and Z represents a halogen atom] has excellent S1P3 antagonist activity.

Description

The present invention relates to a novel diphenyl sulfide derivative or a salt thereof or a hydrate thereof useful as a medicament, and a sphingosine-1-phosphate 3 (S1P3) receptor antagonist and a medicament containing them as an active ingredient.

Sphingosine-1-phosphate (S1P) was considered to be only an intermediate metabolite in sphingosine metabolism. However, it has been reported to have a cell growth promoting effect and a cell motility control effect, and it is clear that this is a new lipid mediator that exhibits various physiological effects such as apoptosis, cell shape regulation, and vasoconstriction. (Non-Patent Document 1, Non-Patent Document 2).

This S1P has two functions as an intracellular second messenger and an intercellular mediator. In particular and studies on the effects of S1P as intercellular mediators been actively plurality of G protein-coupled receptor present on the cell membrane surface (E ndothelial D ifferentiation G ene, EDG) information transmitted via the made (Non-Patent Document 1, Non-Patent Document 3). Currently, five subtypes of Edg-1, Edg-3, Edg-5, Edg-6 and Edg-8 are known for the S1P receptor, and S1P ₁ , S1P ₃ , S1P ₂ , S1P ₄ , S1P, respectively. Also called ₅ .

From various studies on these S1P receptors, it has been reported that so-called S1P receptor modulators exhibiting agonist activity or antagonist activity to this receptor exhibit effectiveness against various diseases. In Patent Document 2 and Non-Patent Documents 4 to 7, S1P3 antagonist is an airway contraction, bronchial asthma, chronic obstructive pulmonary disease (COPD), emphysema, tracheal stenosis, diffuse panbronchiolitis, infection, connective tissue disease or Bronchitis associated with transplantation, diffuse hamartoma pulmonary vascular myomatosis, adult respiratory distress syndrome (ARDS), interstitial pneumonia, lung cancer, hypersensitivity pneumonitis, idiopathic interstitial pneumonia, pulmonary fibrosis, sepsis or It has been reported to be effective as a therapeutic or prophylactic agent for cytokine storm based on influenza virus or RS virus infection.

Patent Documents 3 to 6 show that S1P3 antagonist is arteriosclerosis, intimal thickening, solid tumor, diabetic retinopathy, rheumatoid arthritis, heart failure, ischemic reperfusion injury, cerebral vascular spasm after subarachnoid hemorrhage, coronary Angina pectoris or myocardial infarction caused by vascular spasm, glomerulonephritis, thrombosis, pulmonary edema such as ARDS, cardiac arrhythmia, eye disease, ocular hypertension, glaucoma, glaucomatous retinopathy, optic nerve It is also effective for symptom or macular degeneration.

In addition, there is an activated protein C preparation (rhAPC) as a drug that has been shown to be effective as a therapeutic agent for sepsis, but rhAPC has a bleeding risk as a side effect. Development is desired. Non-Patent Documents 5 and 7 report that the S1P3 receptor is involved in multi-organ failure due to sepsis based on analysis using S1P3 knockout mice, and S1P3 antagonists are effective as a therapeutic or preventive agent for sepsis. It has been suggested that there is. In addition, it has been reported that S1P1 antagonists enhance vascular wall permeability and cause pulmonary edema (Non-patent Document 8). Therefore, in order to obtain a high level of safety for a novel sepsis treatment or prevention drug, the treatment or prevention drug has a weak S1P1 antagonist action, preferably an S1P1 agonist action, and more preferably no action on the S1P1 receptor. It is desired.

As the S1P receptor modulator, for example, the general formula (A) described in Patent Document 1

[Wherein, R ¹ represents a hydrogen atom, a halogen atom, a lower alkyl group having 1 to 4 carbon atoms which may be halogen-substituted, a hydroxy group, a phenyl group, an aralkyl group, a lower alkoxy group having 1 to 4 carbon atoms, trifluoro Methyloxy group, aralkyloxy group which may have a substituent, phenoxy group which may have a substituent, cyclohexylmethyloxy group, aralkyloxy group which may have a substituent, pyridylmethyloxy group, thinner Miloxy group, naphthylmethyloxy group, phenoxymethyl group, hydroxymethyl group, hydroxyethyl group, lower alkylthio group having 1 to 4 carbon atoms, lower alkylsulfinyl group having 1 to 4 carbon atoms, lower alkyl having 1 to 4 carbon atoms sulfonyl group, a benzylthio group, an acetyl group, a nitro group, a cyano group, R ² represents a hydrogen atom, a halogen atom, c Gen substituted a lower alkyl group having 1 to 4 carbon atoms, a lower alkoxy group having 1 to 4 carbon atoms, an aralkyl group, an aralkyl group, R ³ is a hydrogen atom, a halogen atom, a trifluoromethyl group, a carbon A lower alkyl group having 1 to 4 carbon atoms, a lower alkoxy group having 1 to 4 carbon atoms, a hydroxy group, a benzyloxy group, a phenyl group, a lower alkoxymethyl group having 1 to 4 carbon atoms, and a lower alkylthio group having 1 to 4 carbon atoms. R ⁴ represents a hydrogen atom, a halogen atom, a trifluoromethyl group, a lower alkyl group having 1 to 4 carbon atoms, a lower alkoxymethyl group having 1 to 4 carbon atoms, a lower alkylthiomethyl group having 1 to 4 carbon atoms, hydroxymethyl Group, phenyl group, aralkyl group, R ⁵ represents a hydrogen atom, a lower alkyl group having 1 to 4 carbon atoms, X represents O, S, SO, SO ₂ , Y represents —CH ₂ O—, —C H2-, -CH = CH-, -CF = CF-, -CH2CH2-, -CH2CFH-, -CH2CF2-, -CH (OH) CF2- are shown]

A compound represented by the formula is known. However, Patent Document 1 does not include 2-aminophosphate monoester derivatives and 3-aminophosphonic acid derivatives having a diphenyl sulfide skeleton in which an acyl group or a carboxyl group is substituted on the phenyl group. In addition, it is not known that 2-aminophosphate monoester derivatives and 3-aminophosphonic acid derivatives having such a structure exhibit excellent S1P3 receptor antagonist activity.

Patent Document 6 discloses a general formula (B) as an S1P receptor modulator.

[Wherein R ¹ represents a chlorine atom, a linear alkyl group having 1 to 3 carbon atoms or a trifluoromethyl group, R ² represents a fluorine atom or a chlorine atom, and R ³ represents a linear chain having 1 to ³ carbon atoms. An alkyl group, X represents an oxygen atom or a sulfur atom, and n represents an integer of 2 or 3.]

A compound represented by the formula is known. Of the compounds represented by the general formula (B), the general formula (Ba)

[Wherein R ¹ , R ³ , X and n are the same as defined above]

It is reported that the optically active compound represented by the formula has a weak S1P3 agonistic action and an excellent agonistic action on S1P1 and / or S1P4. However, a compound having an asymmetric center opposite to the optically active compound represented by the general formula (Ba) is not known. Moreover, it is not known that such an optically active compound exhibits an excellent S1P3 receptor antagonistic action.

WO04074297 pamphlet WO03020313 pamphlet JP 2005-247691 A WO07043568 pamphlet WO06063033 pamphlet WO08018427 Pamphlet Y. Takuma et al., Mol. Cell. Endocrinol., 177, 3 (2001). Y. Igarashi, Ann, N.Y. Acad. Sci., 845, 19 (1998). H. Okazaki et al., Biochem. Biophs. Res. Commun., 190, 1104 (1993). Y. Gon et.al., Proc Natl Acad Sci U S A. 102 (26), 9270 (2005). F. Nissen et al., Nature, 452,654 (2008) D. Christina et al., Am. J. Pathol., 170 (1), 281 (2007) F. Nissen et al., Blood, 113 (12), 2859 (2009) M.G.Sanna et al., Nature Chemical biology, 2,434 (2006)

The problem to be solved by the present invention is to provide a diphenyl sulfide derivative having excellent S1P3 antagonist activity.

  As a result of intensive studies on S1P3 antagonists, the present inventors have found that a novel diphenyl sulfide derivative has an excellent S1P3 antagonistic action, and completed the present invention.

  That is, the first invention is a general formula (1)

[In Formula (1), R ¹ represents an alkyl group having 1 to 6 carbon atoms or a lower alkoxy group having 1 to 6 carbon atoms which may be substituted with ¹ to 3 halogen atoms, and R ² represents the number of carbon atoms. An alkyl group having 1 to 6 carbon atoms or an alkenyl group having 2 to 6 carbon atoms; R ¹⁰ represents an acyl group having 1 to 6 carbon atoms or a carboxyl group; X represents a methylene or oxygen atom; and Z represents a halogen atom. ]

Or a pharmacologically acceptable salt thereof or a hydrate thereof.

  In the second invention, the compound represented by the general formula (1) is represented by the general formula (1a).

[In formula (1a), R ² and R ¹⁰ are as described above]

The diphenyl sulfide derivative according to the first aspect of the present invention, or a pharmacologically acceptable salt thereof, or a hydrate thereof.

In the third invention, the compound represented by the general formula (1) is
(S) -2-amino-4- {4- (2-carboxy-5-trifluoromethylphenylthio) -2-chlorophenyl} -2-propylbutyl phosphate monoester,
(S) -4- [4- (2-acetyl-5-trifluoromethylphenylthio) -2-chlorophenyl] -2-amino-2-propylbutyl phosphate monoester, or
(S) -3-Amino-5- {4- (2-carboxy-5-trifluoromethylphenylthio) -2-chlorophenyl} -3-propylpentylphosphonic acid according to the first invention It relates to a salt that is physically acceptable or a hydrate thereof.

The fourth invention is a sphingosine-1- containing the diphenyl sulfide derivative according to any one of the first to third inventions or a pharmacologically acceptable salt thereof or a hydrate thereof as an active ingredient. The present invention relates to a medicine based on phosphoric acid 3 (S1P3) receptor antagonistic action.

  The fifth invention also includes airway contraction, bronchial asthma, chronic obstructive pulmonary disease (COPD), emphysema, tracheal stenosis, diffuse panbronchiolitis, infection, connective tissue disease or bronchitis associated with transplantation, diffuse malpractice Pulmonary vascular myomatosis, adult respiratory distress syndrome (ARDS), interstitial pneumonia, lung cancer, hypersensitivity pneumonitis, idiopathic interstitial pneumonia, pulmonary fibrosis, sepsis or cytokines based on influenza virus or RS virus infection The present invention relates to the medicament according to the fourth invention which is a storm treatment or prevention drug.

The sixth invention is caused by arteriosclerosis, intimal thickening, solid tumor, diabetic retinopathy, rheumatoid arthritis, heart failure, ischemic reperfusion injury, cerebral vascular spasm after subarachnoid hemorrhage, coronary vascular spasm Treatment of angina pectoris or myocardial infarction, glomerulonephritis, thrombosis, lung disease caused by pulmonary edema, cardiac arrhythmia, eye disease, ocular hypertension, glaucoma, glaucomatous retinopathy, optic neuropathy or macular degeneration Or it is related with the pharmaceutical of the 4th invention which is a preventive agent.

  The seventh invention relates to the medicament according to the fourth invention, which is a therapeutic or prophylactic agent for sepsis.

  An eighth invention is the diphenyl sulfide derivative according to any one of the first to third inventions, a pharmacologically acceptable salt thereof, or a hydrate thereof, and a pharmacologically acceptable carrier. The present invention relates to a pharmaceutical composition containing

According to the present invention, it is possible to provide a diphenyl sulfide derivative having an excellent S1P3 antagonistic action and S1P3 selectivity. In addition, the diphenyl sulfide derivative of the present invention can be safely used as a medicine because it has little or no hemolytic, tissue damage, or central inhibitory action. Furthermore, the diphenyl sulfide derivative of the present invention is stable in an aqueous solution. The compounds of the present invention having these excellent properties are useful for treating sepsis, airway contraction, bronchial asthma, chronic obstructive pulmonary disease (COPD), emphysema, tracheal stenosis, diffuse panbronchiolitis, infection, connective tissue disease or transplantation. Caused bronchitis, diffuse hamartoma pulmonary vascular myomatosis, adult respiratory distress syndrome (ARDS), interstitial pneumonia, lung cancer, hypersensitivity pneumonitis, idiopathic interstitial pneumonia, pulmonary fibrosis, influenza virus・ Or cytokine storm (overproduction) caused by RS virus infection, arteriosclerosis, intimal thickening, solid tumor, diabetic retinopathy, rheumatoid arthritis, heart failure, ischemic reperfusion injury, brain after subarachnoid hemorrhage Angiogenesis or myocardial infarction caused by vascular spasm, coronary vascular spasm, glomerulonephritis, thrombosis, pulmonary edema caused by pulmonary edema such as ARDS, cardiac arrhythmia, eye disease, ocular hypertension Disease, glaucoma, glaucomatous retinopathy, are useful for the prevention or treatment of optic neuropathy or macular degeneration.

The “halogen atom” in the present invention represents a fluorine atom, a chlorine atom, a bromine atom or an iodine atom. The “C 1-6 alkyl group” is, for example, a linear or i-propyl group such as a methyl group, an ethyl group, an n-propyl group, an n-butyl group, an n-pentyl group or an n-hexyl group. Or the branched C1-C6 hydrocarbon group, such as t-butyl group, is mentioned. Examples of the “C3-C6 cycloalkyl group” include a cyclopropyl group, a cyclobutyl group, a cyclopentyl group, and a cyclohexyl group. Examples of the “aryl group having 6 to 10 carbon atoms” include a phenyl group and a naphthyl group. Examples of the “C 1-6 alkoxy group” include a methoxy group, an ethoxy group, an n-propoxy group, an n-butoxy group, an i-propoxy group, and a t-butoxy group. Examples of the “alkenyl group having 2 to 6 carbon atoms” include an allyl group. Examples of the “acyl group having 1 to 6 carbon atoms” include an acetyl group.

In addition, the “alkyl group having 1 to 6 carbon atoms”, the “alkoxy group having 1 to 6 carbon atoms”, the “acyl group having 1 to 6 carbon atoms”, and the “alkenyl group having 2 to 6 carbon atoms” are substituents. You may have. Examples of the “substituent” include a halogen atom, a trifluoromethyl group, an alkyl group having 1 to 6 carbon atoms, a cycloalkyl group having 3 to 6 carbon atoms, an aryl group having 6 to 10 carbon atoms, and 1 to 6 carbon atoms. An alkoxy group, a benzyloxy group, an acyl group having 1 to 6 carbon atoms, a cyano group, an alkenyl group having 2 to 6 carbon atoms, a hydroxyl group, a nitro group, or an amino group. For example, a trifluoromethyl group etc. are mentioned as "the C1-C6 alkyl group which may be substituted by 1-3 halogen atoms."

For the purpose of obtaining an excellent S1P3 antagonistic action in the present invention, R ¹ is preferably an alkyl group having 1 to 6 carbon atoms which may be substituted with ¹ to 3 halogen atoms, more preferably a trifluoromethyl group. It is. R ² is preferably an alkyl group having 1 to 6 carbon atoms or an alkenyl group having 2 to 6 carbon atoms, more preferably an n-propyl group or an allyl group, and particularly preferably an n-propyl group. R ¹⁰ is preferably an acyl group having 1 to 6 carbon atoms or a carboxyl group, more preferably an acetyl group or a carboxyl group, and particularly preferably a carboxyl group from the viewpoint of safety to living bodies. X is preferably a methylene or oxygen atom, more preferably an oxygen atom. Z is preferably a chlorine atom.

Examples of the pharmacologically acceptable salt in the present invention include acid addition such as hydrochloride, hydrobromide, acetate, trifluoroacetate, methanesulfonate, citrate or tartrate. Examples include salts or alkali addition salts such as sodium salt, potassium salt, calcium salt, magnesium salt or aluminum salt.

According to the present invention, among the compounds represented by the general formula (1), a compound in which X is an oxygen atom, that is, the general formula (1d)

[In formula (1d), R ¹ , R ² , R ¹⁰ and Z are as described above]

Can be produced, for example, by the synthesis route A as shown below.

<Synthesis route A>

In the synthesis route A, the general formula (4)

[In Formula (4), R ³ represents an optionally substituted alkyl group having 1 to 6 carbon atoms, and R ² is as described above.]

The optically active compound represented by general formula (2)

[In formula (2), R ³ is as described above]

An optically active compound represented by the general formula (3)

[In Formula (3), A ^a represents a general leaving group such as ^a halogen atom, a methanesulfonyloxy group, a paratoluenesulfonyloxy group, or a trifluoromethanesulfonyloxy group, and R ² is as described above]

In the presence of a base (step A-1).

  Specifically, first, the compound represented by the general formula (2) is treated with a base at −78 ° C. in a reaction solvent such as 1,4-dioxane, tetrahydrofuran or diethyl ether. Thereafter, the compound represented by the general formula (3) is allowed to act on the anion of the compound represented by the general formula (2) at −78 ° C., and then the temperature is gradually raised to room temperature. To obtain a compound represented by: As the base in the reaction, n-butyllithium or lithium diisopropylamide, preferably n-butyllithium can be used.

  In addition, in this specification, normal temperature means 15-25 degreeC defined by the Japanese Pharmacopoeia.

  In the synthesis route A, the general formula (6)

Wherein (6), A ^b is a halogen atom, a methanesulfonyloxy group, shows a typical leaving group such as p-toluenesulfonyloxy group or a trifluoromethanesulfonyloxy group, R ^2, R ³ and Z are described above Street]

The optically active compound represented by general formula (4) and the optically active compound represented by general formula (5)

Wherein (5), A ^c represents a typical leaving group such as a halogen atom, a methanesulfonyloxy group, p-toluenesulfonyloxy group or a trifluoromethanesulfonyloxy group, A ^b and Z are as described above]

In the presence of a base (step A-2).

Specifically, first, the compound represented by the general formula (4) is treated with a base at −78 ° C. in a reaction solvent such as 1,4-dioxane, tetrahydrofuran or diethyl ether. Thereafter, the compound represented by the general formula (5) is allowed to act at −78 ° C. on the resulting anion of the compound represented by the general formula (4), followed by gradually raising the temperature to room temperature. To obtain a compound represented by: As the base in the reaction, n-butyllithium or lithium diisopropylamide, preferably n-butyllithium can be used.

  In the synthesis route A, the general formula (7)

[In the formula (7), R ⁴ represents a general amino protecting group, and A ^b , R ² , R ³ and Z are as described above]

The compound represented by general formula (6) can be manufactured by acid-hydrolyzing the compound represented by general formula (6), and then protecting the amino group with a general protective reagent. R ⁴ in the formula is not particularly limited as long as it protects the amino group. For example, an acyl group such as an acetyl group or a carbamate such as t-butoxycarbonyl or benzyloxycarbonyl can be used (step A-3). .

Specifically, first, acid hydrolysis is performed at room temperature on the compound represented by the general formula (6) in an inorganic acid or an organic acid or in a mixed solution of an inorganic acid or an organic acid and an organic solvent. At this time, hydrochloric acid or hydrobromic acid can be used as the inorganic acid. As the organic acid, trifluoroacetic acid or the like can be used. As the organic solvent, methanol, ethanol, tetrahydrofuran, 1,4-dioxane, ethyl acetate, or the like can be used. Of these, acid hydrolysis is preferably performed using an aqueous trifluoroacetic acid solution.
Next, after neutralizing with a base to obtain an aminoester compound, the aminoester compound and acid chloride or acid anhydride are reacted in a solvent at 0 ° C. to room temperature, and represented by the general formula (7). To obtain the compound. At this time, ethyl acetate, tetrahydrofuran, N, N-dimethylformamide, 1,4-dioxane, methylene chloride, chloroform, methanol, ethanol, acetonitrile, or the like can be used as the solvent. As the acid chloride, acetyl chloride or benzyloxycarbonyl chloride can be used. As the acid anhydride, acetic anhydride or di-t-butyl dicarbonate can be used. Of these, it is preferable to carry out the reaction using di-t-butyl dicarbonate.

In the synthesis route A, the general formula (8)

[In the formula (8), A ^b , R ² , R ⁴ and Z are as described above]

Can be produced by reducing the compound represented by the general formula (7) (step A-4).

For example, in a reaction solvent such as tetrahydrofuran, 1,4-dioxane, ethanol or methanol, the compound represented by the general formula (7) is reduced at a temperature of 0 ° C. to heating under reflux, preferably normal temperature, using a reducing agent. . Reducing agents include borane or alkylborane derivatives such as 9-borabicyclo [3.3.1] nonane (9-BBN), diisobutylaluminum hydride ((iBu) ₂ AlH), sodium borohydride (NaBH ₄ ), hydrogenation Metal hydride complex compounds such as lithium boron (LiBH ₄ ) or lithium aluminum hydride (LiAlH ₄ ) can be used. Preferably, the reducing agent is lithium borohydride.

In the synthesis route A, the general formula (10)

[In the formula (10), R ¹¹ represents an acyl group having 1 to 6 carbon atoms or a general formula (31)

(In the formula (31), R ¹² represents an alkyl group having 1 to 6 carbon atoms)
Wherein R ¹ , R ² , R ⁴ and Z are as described above]

The compound represented by general formula (8) and the compound represented by general formula (9)

[In formula (9), R ¹ and R ¹¹ are as described above]

It can manufacture by making the compound represented by these react (process A-5).
For example, the reaction is carried out at room temperature to heating reflux using a catalyst in a reaction solvent such as toluene, N, N-dimethylformamide, 1,4-dioxane, tetrahydrofuran or diethyl ether in the presence of an inorganic base or an organic base. Can be done at temperature. As the inorganic base, sodium carbonate or potassium t-butoxide can be used. As the organic base, diisopropylethylamine or the like can be used. The catalyst may be a palladium compound such as tris (dibenzylideneacetone) dipalladium (0) or palladium (II) acetate, preferably tris (dibenzylideneacetone) dipalladium (0).
Also, phosphines such as 4,5-bis (diphenylphosphino) -9,9-dimethylxanthene, bis [2- (diphenylphosphino) phenyl] ether, 1,1′-bis (dit-butylphosphino) ferrocene The compound can also be added to the reaction solvent as a reaction accelerator.

In the synthesis route A, the general formula (12)

[In formula (12), R ¹ , R ² , R ³ , R ⁴ , R ¹¹ and Z are as described above]

The compound represented by general formula (10) and the compound represented by general formula (11)

[In formula (11), R ³ is as described above]

It can manufacture by making the compound represented by these react (process A-6).

For example, the reaction can be performed at 0 ° C. to room temperature in the presence of carbon tetrabromide and pyridine using no solvent or methylene chloride, chloroform, acetonitrile, ethyl acetate, tetrahydrofuran, or diethyl ether as a solvent.

Among the compounds represented by the general formula (1d) in the synthesis route A, R ¹⁰ is an acyl group having 1 to 6 carbon atoms, and among the compounds represented by the general formula (12), R ¹¹ has 1 carbon atom. Compounds that are ˜6 acyl groups can be prepared by acid hydrolysis and / or treatment with nucleophiles such as trimethylsilyl bromide or trimethylsilyl iodide. In addition, among the compounds represented by the general formula (1d) in the synthesis route A, R ¹⁰ is a carboxyl group, and among the compounds represented by the general formula (12), R ¹¹ is represented by the general formula (31). The compound which is an ester group can be produced by acid hydrolysis and / or treatment with a nucleophile such as trimethylsilyl bromide or trimethylsilyl iodide, and then, if necessary, alkaline hydrolysis (Step A). -7).

  In the case of the acid hydrolysis reaction, the reaction can be carried out at a heating reflux temperature in an inorganic acid such as hydrochloric acid or hydrobromic acid, or in a mixed solution of an organic solvent such as methanol or ethanol and an inorganic acid. In the treatment using a nucleophile, acetonitrile or methylene chloride may be used as a preferred reaction solvent, and trimethylsilyl bromide or trimethylsilyl iodide may be allowed to act at 0 ° C. to room temperature. Alternatively, the treatment using a nucleophilic reagent can also be performed by acting a combination of trimethylsilyl chloride and sodium bromide or trimethylsilyl chloride and sodium iodide.

The alkaline hydrolysis reaction can be carried out by a method usually used for ester hydrolysis. For example, the reaction can be performed at a temperature from room temperature to reflux using a sodium hydroxide aqueous solution, a potassium hydroxide aqueous solution, or a lithium hydroxide aqueous solution in a solvent such as methanol, ethanol, or tetrahydrofuran.

The compound represented by the general formula (7) in the synthetic route A can also be produced, for example, by the synthetic route B as shown below.

<Synthetic route B>

In the synthesis route B, the general formula (14)

[In formula (14), A ^b , R ³ and Z are as described above]

The optically active compound represented by the general formula (13)

[In formula (13), R ³ is as described above]

It can manufacture with the method similar to process A-2 using the optically active compound represented by and the compound represented by General formula (5) (process B-1).

  In the synthesis route B, the general formula (15)

[In formula (15), A ^b , R ² , R ³ and Z are as described above]

The optically active compound represented by general formula (14) and the compound represented by general formula (3) are manufactured by the method similar to process A-1. (Step B-2).

The compound represented by general formula (7) in the synthetic pathway B can be manufactured by the method similar to process A-3 using the compound represented by general formula (15) (process B-3).

The compound represented by the general formula (10) in the synthesis route A can also be produced, for example, by the synthesis route C as shown below.

<Synthetic route C>

In the synthesis route C, the general formula (17)

[In formula (17), R ¹ , R ² , R ³ , R ¹¹ and Z are as described above]

The optically active compound represented by general formula (4) and the optically active compound represented by general formula (16)

[In formula (16), R ¹ , R ¹¹ , ^Ac and Z are as described above]

It can manufacture by the method similar to process A-2 using the compound represented by (process C-1).

  In the synthesis route C, the general formula (18)

[In formula (18), R ¹ , R ² , R ³ , R ⁴ , R ¹¹ and Z are as described above]

The compound represented by this can be manufactured by the method similar to process A-3 using the compound represented by General formula (17) (process C-2).

The compound represented by general formula (10) in the synthetic pathway C can be manufactured by the method similar to process A-4 using the compound represented by general formula (18) (process C-3).

The compound represented by the general formula (18) in the synthesis route C can also be produced, for example, by the synthesis route D as shown below.

<Synthesis route D>

In the synthesis route D, the general formula (19)

[In formula (19), R ¹ , R ³ , R ¹¹ and Z are as described above]

The optically active compound represented by the formula is produced by the same method as in step A-2 using the optically active compound represented by the general formula (13) and the compound represented by the general formula (16). (Step D-1).

In the synthesis route D, the general formula (20)

[In the formula (20), R ¹ , R ² , R ³ , R ¹¹ and Z are as described above]

The optically active compound represented by general formula (19) and the compound represented by general formula (3) are manufactured by the method similar to process A-1. (Step D-2).

The compound represented by general formula (18) in the synthetic pathway D can be manufactured by the method similar to process A-3 using the compound represented by general formula (20) (process D-3).

Of the compounds represented by the general formula (1), a compound in which X is methylene, that is, the general formula (1e)

[In formula (1e), R ¹ , R ² , R ¹⁰ and Z are as described above]

Can be produced by, for example, the synthesis route E as shown below.

<Synthesis route E>

In the synthesis route E, the general formula (26)

[In formula (26), R ¹ , R ² , R ⁴ , R ¹¹ and Z are as described above.]

Can be produced by oxidizing the compound represented by the general formula (10) (step F-1).
For the reaction, a generally used oxidation method of alcohol to aldehyde can be used. For example, oxidation treatment using a chromium oxide-pyridine complex such as pyridinium chlorochromate or pyridinium dichromate, or oxidation using hypervalent iodine such as Dess-Martin oxidation can be given. Alternatively, dimethyl sulfoxide oxidation using various dimethyl sulfoxide activators such as oxalyl chloride, trifluoroacetic anhydride, acetic anhydride, dicyclohexylcarbodiimide, and sulfur trioxide-pyridine complex may also be mentioned.

In the synthesis route E, the general formula (29)

[In formula (29), R ¹ , R ² , R ³ , R ⁴ , R ¹¹ and Z are as described above.]

Examples of the compound represented by general formula (26) and the general formula (27)

[In formula (27), R ³ is as described above]

Can be produced by reacting in a reaction solvent in the presence of a base (Step F-2).

As the base in the reaction, sodium hydride, potassium hydride, sodium alkoxide, potassium alkoxide, n-butyllithium, or the like, preferably n-butyllithium can be used. As the reaction solvent, tetrahydrofuran, diethyl ether, 1,4-dioxane, or the like can be used. Moreover, reaction temperature can be made into -78 degreeC-normal temperature.

  In the synthesis route E, the general formula (30)

[In formula (30), R ¹ , R ² , R ³ , R ⁴ , R ¹¹ and Z are as described above.]

Can be produced by reducing the compound represented by formula (29) (step F-3).

For example, the reaction can be carried out in the presence of a catalytic reduction catalyst in a solvent such as ethanol, methanol, tetrahydrofuran, N, N-dimethylformamide or ethyl acetate at normal temperature to pressurized hydrogen pressure at room temperature. As the catalytic reduction catalyst, palladium carbon, platinum carbon, platinum oxide, rhodium carbon, ruthenium carbon, or the like can be used.

The reaction can also be performed by diimide reduction. For example, it can be carried out using potassium azodicarboxylate in a solvent such as pyridine, ethanol, methanol, dimethyl sulfoxide, 1,4-dioxane in the presence of acetic acid at a temperature from normal temperature to heating reflux.

The compound represented by general formula (1e) in the synthetic pathway E can be manufactured by the method similar to process A-7 using the compound represented by general formula (30) (process F-4).

In addition, about the synthesis method of the compound represented by General formula (16), it can manufacture by the method described in each pamphlet of WO03029184, WO03029205, WO04026817, WO04074297, and WO050444780.

The diphenyl sulfide derivative of the present invention or a pharmacologically acceptable salt thereof or a hydrate thereof exhibits an excellent S1P3 antagonistic action. Therefore, a pharmaceutical comprising at least one of these as an active ingredient is effective as a therapeutic or prophylactic agent for a disease for which an S1P3 antagonist is known to be effective as a therapeutic or prophylactic agent. Diseases for which the S1P3 antagonist is known to be effective as a therapeutic or prophylactic agent include, for example, sepsis, airway constriction, bronchial asthma, chronic obstructive pulmonary disease (COPD), emphysema, tracheal stenosis, diffuse generalization Bronchitis, infection, connective tissue disease or bronchitis associated with transplantation, diffuse hamartoma pulmonary vascular myomatosis, adult respiratory distress syndrome (ARDS), interstitial pneumonia, lung cancer, hypersensitivity pneumonitis, idiopathic stroma And pneumonia, pulmonary fibrosis, influenza virus, and cytokine storm based on RS virus infection.

In addition to the above diseases, the medicament of the present invention is also effective for treating or preventing diseases that are known to have an effective S1P3 antagonistic action. Diseases for which the S1P3 antagonist action is known to be effective include, for example, arteriosclerosis, intimal thickening, solid tumors, diabetic retinopathy, rheumatoid arthritis, heart failure, ischemic reperfusion injury, subarachnoid hemorrhage Later cerebral vascular spasm, angina or myocardial infarction caused by coronary vascular spasm, glomerulonephritis, thrombosis, pulmonary edema such as ARDS, cardiac arrhythmia, eye disease, ocular hypertension, glaucoma Glaucomatous retinopathy, optic neuropathy or macular degeneration.

The medicament of the present invention can be administered orally or by parenteral means such as rectal, subcutaneous, intravenous, intramuscular or transdermal.

In order to use the compound of the present invention, a pharmacologically acceptable salt thereof or a hydrate thereof as a pharmaceutical, it may be in the form of a solid composition, a liquid composition or other composition, and if necessary The best one is selected. The pharmaceutical composition of the present invention can also be produced by blending the compound of the present invention with a pharmacologically acceptable carrier. Specifically, conventional excipients, extenders, binders, disintegrants, coating agents, sugar coatings, pH adjusters, solubilizers or aqueous or non-aqueous solvents are added, and tablets are prepared by conventional formulation techniques. , Pills, capsules, granules, powders, powders, solutions, emulsions, suspensions or injections.

Next, the present invention will be described with reference to specific examples, but the present invention is not limited to these examples.

<Reference Example 1>
(2R, 5R) -2- (4-Bromo-2-chlorophenyl) ethyl-3,6-dimethoxy-5-isopropyl-2-propyl-2,5-dihydropyrazine

N-Butyllithium-hexane in a solution of (5R) -3,6-dimethoxy-2-propyl-5-isopropyl-2,5-dihydropyrazine (5.21 g) in tetrahydrofuran (100 mL) at −78 ° C. under an argon atmosphere A solution (1.60 mol / L, 15.1 mL) was added to prepare a reaction solution. Next, the reaction solution was stirred at −78 ° C. for 30 minutes. Further, a solution of 4-bromo-2-chloro-1- (2-iodoethyl) benzene (9.54 g) in tetrahydrofuran (15 mL) was added to the reaction mixture, and the mixture was stirred at −78 ° C. for 30 minutes and then at 0 ° C. for 1 hour. Water was added to the reaction mixture, and the mixture was extracted with ethyl acetate, washed successively with water and saturated brine, and dried over anhydrous sodium sulfate. After anhydrous sodium sulfate was removed by filtration, the solvent was distilled off under reduced pressure, and the residue was purified using silica gel column chromatography (hexane: ethyl acetate = 60: 1) to obtain the desired product (8.01 g) as a colorless oil.
¹ H NMR (CDCl ₃ , 400 MHz): δ 0.70 (3H, d, J = 6.7 Hz), 0.86 (3H, t, J = 7.3 Hz), 1.11 (3H, d, J = 6.7 Hz), 1.15- 1.30 (2H, m), 1.49-1.62 (1H, m), 1.71-1.84 (2H, m), 1.98 (1H, td, J = 12.4, 4.8 Hz), 2.29-2.47 (3H, m), 3.69 ( 3H, s), 3.70 (3H, s), 3.95 (1H, d, J = 3.0 Hz), 7.01 (1H, d, J = 7.9 Hz), 7.27 (1H, dd, J = 7.9, 1.8 Hz), 7.46 (1H, d, J = 1.8 Hz).
ESIMS (+): 443 [M + H] ⁺ .

<Reference Example 2>
(S) -4- (4-Bromo-2-chlorophenyl) -2-t-butoxycarbonylamino-2-propylbutyric acid methyl ester

A 50% trifluoroacetic acid-water solution (800 mL) was added to the compound of Reference Example 1 (53.4 g), stirred at room temperature for 1 hour, and then allowed to stand overnight at room temperature. The mixture was neutralized with saturated aqueous sodium hydrogen carbonate solution and extracted with ethyl acetate. The extract was washed with water and saturated brine, and dried over anhydrous sodium sulfate. After concentration of the extract, the residue was dissolved in acetonitrile (375 mL), and di-tert-butoxy dicarbonate (19.6 g) was added. The mixture was stirred at room temperature for 1 hour and left at room temperature overnight. Water was added to the reaction mixture, and the mixture was extracted with ethyl acetate, washed successively with water and saturated brine, and dried over anhydrous sodium sulfate. The solvent was distilled off under reduced pressure, and the residue was purified by silica gel column chromatography (hexane: ethyl acetate = 6: 1) to obtain the desired product (35.6 g) as a colorless oil.
¹ H NMR (CDCl ₃ , 400 MHz): δ 0.89 (3H, t, J = 7.3 Hz) .0.96-1.10 (1H, m), 1.25-1.39 (1H, m), 1.46 (9H, s), 1.69 (1H, ddd, J = 13.9, 11.5. 4.8 Hz), 1.99-2.10 (1H, m), 2.20-2.35 (1H, m), 2.42 (1H, ddd, J = 13.9, 11.5, 4.8 Hz), 2.49 -2.60 (1H, m), 2.64 (1H, td, J = 13.9, 4.8 Hz), 3.74 (3H, s), 5.62 (1H, br s), 7.03 (1H, d, J = 8.5 Hz), 7.29 (1H, dd, J = 8.5, 1.8 Hz), 7.48 (1H, J = 1.8 Hz).
ESIMS (+): 448 [M + H] ⁺ .

<Reference Example 3>
(R) -2- [2- (4-Bromo-2-chlorophenyl) ethyl] -2-t-butoxycarbonylaminopentan-1-ol

To a solution of the compound of Reference Example 2 (35.6 g) in tetrahydrofuran (250 mL) was added lithium borohydride-tetrahydrofuran solution (3 mol / L, 125 mL) under ice-cooling, and ethanol (37.5 mL) was added dropwise. The mixture was stirred for 2 hours under ice cooling. To the reaction solution was added 10% aqueous citric acid solution, and the mixture was extracted with ethyl acetate, washed with water and then saturated brine, and dried over anhydrous sodium sulfate. The solvent was distilled off under reduced pressure, and the residue was purified by silica gel column chromatography (hexane: ethyl acetate = 2: 1) to obtain the desired product (28.6 g) as a colorless solid.
¹ H NMR (CDCl ₃ , 400 MHz): δ 0.96 (3H, t, J = 7.3 Hz), 1.29-1.42 (2H, m), 1.44 (9H, s), 1.53-1.62 (2H, m), 1.81 (1H, ddd, J = 13.9, 11.5, 5.4 Hz), 1.93 (1H, ddd, J = 13.9, 11.5, 5.4 Hz), 2.59-2.75 (2H, m), 3.73 (2H, d, J = 6.7 Hz ), 4.15 (1H, br s), 4.62 (1H, br s), 7.11 (1H, d, J = 7.9 Hz), 7.31 (1H, dd, J = 7.9, 1.8 Hz), 7.49 (1H, d, J = 1.8 Hz).
ESIMS (+): 420 [M + H] ⁺ .

<Reference Example 4>
2-hydroxy-4-trifluoromethyl methyl benzoate

To a solution of 2-hydroxy-4-trifluoromethylbenzoic acid (24.0 g) in methanol (600 mL) was added thionyl chloride (41.5 g) under ice cooling. After stirring for 7 hours under heating to reflux, the reaction solution was concentrated under reduced pressure, and the resulting residue was purified by silica gel column chromatography (Fuji Silysia Chemical Chromathex NH, ethyl acetate) to obtain the desired product (25.9 g) as colorless. Obtained as an oil.
¹ H NMR (CDCl ₃ , 400 MHz): δ 3.99 (3H, s), 7.12 (1H, dd, J = 8.6, 1.2 Hz), 7.25 (1H, d, J = 8.6 Hz).
EIMS (+): 220 [M + H] ⁺ .

<Reference Example 5>
2-Dimethylcarbamothioyloxy-4-trifluoromethyl methyl benzoate

To a solution of the compound of Reference Example 4 (20.0 g) in N, N-dimethylformamide (100 mL) was added 1,5-diazabicyclo [5.4.0] undec-7-ene (34.6 g) and dimethylthiocarbamoyl chloride (22.5 g). ) And stirred at room temperature for 7 hours. The reaction mixture was poured into water and extracted with ethyl acetate. The organic layer was washed with saturated brine, dried over anhydrous magnesium sulfate, and the solvent was distilled off under reduced pressure. The obtained solid was suspended in diisopropyl ether and collected by filtration to obtain the desired product (20.3 g) as a yellow solid.
¹ H NMR (CDCl ₃ , 400 MHz): δ3.41 (3H, s), 3.46 (3H, s), 3.87 (3H, s), 7.40 (1H, d, J = 1.2 Hz), 7.56 (1H, dd, J = 8.6, 1.2 Hz), 8.10 (1H, d, J = 8.6 Hz).
EIMS (+): 307 [M + H] ⁺ .

<Reference Example 6>
Methyl 2-dimethylcarbamoylthio-4-trifluoromethylbenzoate

The compound of Reference Example 5 (10.0 g) was stirred at 190 ° C. without solvent for 4 hours. The reaction product was cooled to room temperature, and the obtained residue was purified by silica gel column chromatography (hexane: ethyl acetate = 2: 1) to obtain the desired product (6.0 g) as a colorless oil.
¹ H NMR (CDCl ₃ , 400 MHz): δ 3.03 (3H, br), 3.14 (3H, br), 3.91 (3H, s), 7.67 (1H, dd, J = 8.6, 1.2 Hz), 7.87 (1H , d, J = 1.2 Hz), 7.98 (1H, d, J = 8.6 Hz).
EIMS (+): 307 [M + H] ⁺ .

<Reference Example 7>
2-Mercapto-4-trifluoromethylbenzoic acid methyl ester

A 25% sodium methoxide-methanol solution (19.5 mL) was added to the compound of Reference Example 6 (2.00 g), and the mixture was stirred at 60 ° C. for 30 minutes. The reaction solution was cooled to room temperature, and 6 mol / L hydrochloric acid was added to adjust pH <1. Extraction was performed with diethyl ether, and the organic layer was extracted with 0.5 mol / L aqueous sodium hydroxide. The extracted aqueous layer was washed with diethyl ether and adjusted to pH <1 with concentrated hydrochloric acid. The aqueous layer was extracted with diethyl ether, dried over anhydrous magnesium sulfate, and the solvent was evaporated under reduced pressure to obtain the desired product (1.23 g) as a colorless oil.
¹ H NMR (CDCl ₃ , 400 MHz): δ 3.96 (3H, s), 4.92 (1H, s), 7.38 (1H, dd, J = 8.6, 1.2 Hz), 7.57 (1H, d, J = 1.2 Hz ), 8.11 (1H, d, J = 8.6 Hz).
EIMS (+): 236 [M + H] ⁺ .

<Reference Example 8>
2-Iodo-5- (trifluoromethyl) phenol

60% sodium hydride (4.93 g) was added to a toluene (200 mL) solution of 3- (trifluoromethyl) phenol (20.0 g), and the mixture was stirred at room temperature for 30 minutes. Then iodine (31.2 g) was added and stirred at room temperature for 1 hour. 3 mol% hydrochloric acid and ethyl acetate were added to the reaction solution to separate the layers. The organic layer was washed with saturated aqueous sodium thiosulfate solution and saturated brine, dried over anhydrous magnesium sulfate, and the solvent was evaporated under reduced pressure. The obtained residue was purified by silica gel column chromatography (hexane: ethyl acetate = 8: 1) to obtain the desired product (22.6 g) as a colorless oil.
¹ H NMR (CDCl ₃ , 400 MHz): δ 5.59 (1H, s), 6.93 (1H, dd, J = 8.6, 1.2 Hz), 7.22 (1H, d, J = 1.2 Hz), 7.78 (1H, d , J = 8.6 Hz).
EIMS (+): 287 [M] ⁺ .

<Reference Example 9>
1- {2-hydroxy-4- (trifluoromethyl) phenyl} ethanone

To a solution of the compound of Reference Example 8 (15.0 g) in N, N-dimethylformamide (80 mL), triethylamine (7.91 g), n-butyl vinyl ether (26.1 g), 1,3-bis (diphenylphosphino) propane ( 4.29 g) and palladium acetate (1.17 g) were added, and the mixture was stirred at 110 ° C. for 5 hours. The reaction solution was cooled to room temperature, and water and ethyl acetate were added for liquid separation. The organic layer was washed with saturated brine, dried over anhydrous magnesium sulfate, and the solvent was distilled off under reduced pressure. The obtained residue was purified by silica gel column chromatography (hexane: ethyl acetate = 30: 1) to obtain the desired product (6.79 g) as a colorless oil.
¹ H NMR (CDCl ₃ , 400 MHz): δ 2.69 (3H, s), 7.15 (1H, dd, J = 8.6, 1.2 Hz), 7.26 (1H, d, J = 1.2 Hz), 7.86 (1H, d , J = 8.6 Hz).
EIMS (+): 204 [M + H] ⁺ .

<Reference Example 10>
Dimethylthiocarbamic acid O- {2-acetyl-5- (trifluoromethyl) phenyl}

Using the compound of Reference Example 9 (3.50 g), the reaction was carried out in the same manner as in Reference Example 5 to obtain the desired product (4.55 g) as a yellow oil.
¹ H NMR (CDCl ₃ , 400 MHz): δ 2.58 (3H, s), 3.42 (3H, sH, h), 3.46 (3H, s), 7.37 (1H, d, J = 1.2 Hz), 7.57 (1H , dd, J = 8.6, 1.2 Hz), 7.85 (1H, d, J = 8.6 Hz).
EIMS (+): 291 [M + H] ⁺ .

<Reference Example 11>
Dimethylthiocarbamic acid S- {2-acetyl-5- (trifluoromethyl) phenyl}

The reaction was conducted in the same manner as in Reference Example 6 using the compound of Reference Example 10 (4.50 g) to obtain the desired product (3.03 g) as a colorless oil.
¹ H NMR (CDCl ₃ , 400 MHz): δ 2.61 (3H, s), 3.02 (3H, br), 3.11 (3H, br), 7.63-7.70 (2H, m), 7.83 (1H, s).
EIMS (+): 291 [M + H] ⁺ .

<Reference Example 12>
1- {2-Mercapto-4- (trifluoromethyl) phenyl} ethanone

Using the compound of Reference Example 11 (400 mg), the reaction was carried out in the same manner as in Reference Example 7 to obtain the desired product (210 mg) as a colorless oil.
¹ H NMR (CDCl ₃ , 400 MHz): δ 2.67 (3H, s), 4.61 (1H, br), 7.44 (1H, dd, J = 8.6, 1.2 Hz), 7.56 (1H, d, J = 1.2 Hz ), 7.97 (1H, d, J = 8.6 Hz).
EIMS (+): 220 [M + H] ⁺ .

<Reference Example 13>
(2-Methoxy-5-trifluoromethylphenylthio) ethoxymethane-1-thione

Water (13 mL) and concentrated hydrochloric acid (6.9 mL) were added to a solution of 2-methoxy-5-trifluoromethylaniline (2.50 g) in methanol (13 mL) under an argon atmosphere and ice cooling. After stirring for 10 minutes, sodium nitrite (1.26 g) was added to the reaction mixture, and the mixture was stirred for 1 hour under ice cooling. The reaction solution was slowly added dropwise to an aqueous solution (13 mL) of potassium ethylxanthate (4.19 g) heated to 65 ° C., and stirred at 65 ° C. for 1 hour. The reaction solution was returned to room temperature, ice water was added to the reaction solution, extracted with ethyl acetate, washed in turn with water and saturated brine, and then dried over anhydrous sodium sulfate. The solvent was distilled off under reduced pressure and the residue was purified by silica gel column chromatography (hexane: ethyl acetate = 20: 1) to obtain the desired product (913 mg) as a colorless oil.
¹ H NMR (CDCl ₃ , 400 MHz): δ 1.31 (3H, t, J = 7.3 Hz) .3.92 (3H, s), 4.60 (2H, q, J = 7.3 Hz), 7.04 (1H, d, J = 8.6 Hz), 7.71 (1H, dd, J = 8.6, 2.4 Hz), 7.73 (1H, d, J = 2.4 Hz).
EIMS (+): 296 [M] ⁺ .

<Reference Example 14>
2-methoxy-5-trifluoromethylbenzenethiol

Lithium aluminum hydride (5.92 g) was added to a solution of the compound of Reference Example 13 (38.4 g) in diethyl ether (500 mL) under an argon atmosphere and ice cooling, and the mixture was stirred for 30 minutes under ice cooling. 1 mol / L hydrochloric acid was added to the reaction solution, and the mixture was extracted with diethyl ether. The organic layer was extracted with a 1 mol / L aqueous sodium hydroxide solution, the aqueous layer was washed with diethyl ether and adjusted to pH 2 with concentrated hydrochloric acid. The aqueous layer was extracted with diethyl ether, washed successively with water and saturated brine, and dried over anhydrous magnesium sulfate. The solvent was distilled off under reduced pressure to obtain the desired product (400 mg) as a colorless oil.
¹ H NMR (CDCl ₃ , 400 MHz): δ3.93 (1H, s), 3.95 (3H, s), 6.90 (1H, d, J = 8.6 Hz), 7.38 (1H, dd, J = 8.6, 2.4 Hz), 7.51 (1H, d, J = 2.4 Hz).
EIMS (+): 296 [M] ⁺ .

<Reference Example 15>
(S) -2-t-butoxycarbonylamino-4- {2-chloro-4- (2-methoxycarbonyl-5-trifluoromethylphenylthio) phenyl} -2-propylbutan-1-ol

Under an argon gas atmosphere, the compound of Reference Example 3 (500 mg), the compound of Reference Example 7 (337 mg), tris (dibenzylideneacetone) dipalladium (109 mg) and 4,5-bis (diphenylphosphino) -9 , 9-Dimethylxanthene (138 mg) was dissolved in dioxane (8 mL), diisopropylethylamine (0.42 mL) was added, and the mixture was stirred with heating under reflux for 2 hr. Water was added to the reaction mixture, and the mixture was extracted with ethyl acetate. The organic layer was washed with saturated brine and dried over anhydrous sodium sulfate. After evaporating the solvent under reduced pressure, the residue was purified by silica gel column chromatography (hexane: ethyl acetate = 4: 1) to obtain the desired product (638 mg) as a brown oil.
¹ H NMR (CDCl ₃ , 400 MHz): δ0.98 (3H, t, J = 7.3 Hz), 1.33-1.63 (4H, m), 1.45 (9H, s), 1.85-1.92 (1H, m), 1.97-2.05 (1H, m), 2.75-2.80 (2H, m), 3.77 (2H, d, J = 6.1 Hz), 3.98 (3H, s), 4.19 (1H, brs), 4.66 (1H, brs) , 7.06 (1H, s), 7.34-7.39 (3H, m), 7.55 (1H, d, J = 1.8 Hz), 8.08 (1H, d, J = 8.6 Hz).
ESIMS (+): 576 [M + H] ⁺ .

<Reference Example 16>
(S) -2-t-butoxycarbonylamino-4- {2-chloro-4- (2-methoxycarbonyl-5-trifluoromethylphenylthio) phenyl} -1-dimethoxyphosphoryloxy-2-propylbutane

Dissolve the compound of Reference Example 15 (598 mg) in pyridine (5 mL), add carbon tetrabromide (689 mg) and trimethyl phosphite (0.245 mL) under ice cooling, and stir at room temperature for 4 hours. did. Water was added to the reaction mixture, and the mixture was extracted with ethyl acetate. The organic layer was washed with saturated brine and dried over anhydrous sodium sulfate. After evaporating the solvent under reduced pressure, the residue was purified by silica gel column chromatography (hexane: ethyl acetate = 1: 1) to obtain the desired product (504 mg) as a yellow oil.
¹ H NMR (CDCl ₃ , 400 MHz): δ0.97 (3H, t, J = 7.3 Hz), 1.36-1.45 (2H, m), 1.45 (9H, s), 1.63-1.72 (2H, m), 1.82-1.88 (1H, m), 2.05-2.09 (1H, m), 2.75-2.79 (2H, m), 3.78 (3H, d, J = 1.2 Hz), 3.81 (3H, d, J = 1.2 Hz) , 3.98 (3H, s), 4.13-4.16 (1H, m), 4.25-4.29 (1H, m), 4.55 (1H, brs), 7.05 (1H, s), 7.31-7.39 (3H, m), 7.55 (1H, d, J = 1.2 Hz), 8.08 (1H, d, J = 8.6 Hz).
ESIMS (+): 684 [M + H] ⁺ .

(S) -2-Amino-4- {4- (2-carboxy-5-trifluoromethylphenylthio) -2-chlorophenyl} -2-propylbutyl phosphate monoester

The compound of Reference Example 16 (500 mg) was dissolved in a 10% hydrochloric acid-methanol solution (15 mL) and stirred at room temperature for 19 hours and at 50 ° C. for 2 hours. After evaporating the solvent under reduced pressure, the residue was dissolved in acetonitrile (7 mL), iodotrimethylsilane (0.44 mL) was added under ice cooling, and the mixture was stirred at the same temperature for 2 hr. Water was added to the reaction solution, and the resulting solid was collected by filtration, washed with water and acetonitrile, and dried at 60 ° C. under reduced pressure. The obtained solid was dissolved in methanol (5 mL), 1 mol / L aqueous sodium hydroxide solution (1.7 mL) was added, and the mixture was stirred at room temperature for 14 hr. After distilling off the solvent under reduced pressure, the residue was diluted with water and acidified (pH 2-3) with 1 mol / L hydrochloric acid under ice cooling. The resulting solid was collected by filtration to give the object product (290 mg) as a colorless solid.
¹ H NMR (DMSO-d ₆ , 400 MHz): δ0.90 (3H, t, J = 7.3 Hz), 1.34-1.36 (2H, m), 1.60-1.62 (2H, m), 1.80-1.82 (2H , m), 2.74-2.76 (2H, m), 3.80-3.90 (2H, m), 6.94 (1H, s), 7.44 (1H, dd, J = 8.0, 1.2 Hz), 7.52 (1H, d, J = 8.0 Hz), 7.56 (1H, d, J = 8.0 Hz), 7.61 (1H, d, J = 1.2 Hz), 8.08 (1H, d, J = 8.0 Hz).
HRESIMS (+): 542.07781 (calculated as C ₂₁ H ₂₅ ClF ₃ NO ₆ PS 542.07808).

<Reference Example 17>
(S) -4- [4- (2-acetyl-5-trifluoromethylphenylthio) -2-chlorophenyl] -2-t-butoxycarbonylamino-2-propylbutan-1-ol

Using the compound of Reference Example 3 (375 mg) and the compound of Reference Example 12 (251 mg), the reaction was carried out in the same manner as in Reference Example 15 to obtain the desired product (450 mg) as a colorless oil.
¹ H NMR (CDCl ₃ , 400 MHz): δ0.98 (3H, t, J = 6.7 Hz), 1.30-1.43 (2H, m), 1.44 (9H, s), 1.53-1.60 (2H, m), 1.83-1.91 (1H, m), 1.93-2.01 (1H, m), 2.69 (3H, s), 2.74-2.83 (2H, m), 3.76 (2H, d, J = 6.7 Hz), 4.18 (1H, brs), 4.66 (1H, brs), 7.13-7.14 (1H, m), 7.31-7.33 (2H, m), 7.43-7.45 (1H, m), 7.50-7.52 (1H, m), 7.88-7.92 ( 1H, m).
ESIMS (+): 560 [M + H] ⁺ .

<Reference Example 18>
(S) -4- [4- (2-acetyl-5-trifluoromethylphenylthio) -2-chlorophenyl] -2-t-butoxycarbonylamino-1-dimethoxyphosphoryloxy-2-propylbutane

Using the compound of Reference Example 17 (479 mg), the reaction was carried out in the same manner as in Reference Example 16 to obtain the desired product (392 mg) as a yellow oil.
¹ H NMR (CDCl ₃ , 400 MHz): δ0.97 (3H, t, J = 7.3 Hz), 1.34-1.42 (2H, m), 1.45 (9H, s), 1.64-1.70 (2H, m), 1.82-1.88 (2H, m), 2.69 (3H, s), 2.77 (2H, t, J = 8.6 Hz), 3.78 (3H, d, J = 1.2 Hz), 3.80 (3H, d, J = 1.2 Hz ), 4.13-4.15 (1H, m), 4.25-4.29 (1H, m), 4.53 (1H, brs), 7.13 (1H, s), 7.29-7.34 (2H, m), 7.43 (1H, dd, J = 8.0, 1.2 Hz), 7.51 (1H, d, J = 1.2 Hz), 7.90 (1H, d, J = 8.0 Hz).
ESIMS (+): 668 [M + H] ⁺ .

 (S) -4- [4- (2-acetyl-5-trifluoromethylphenylthio) -2-chlorophenyl] -2-amino-2-propylbutyl phosphate monoester

The compound of Reference Example 18 (390 mg) was dissolved in 4 mol / L hydrochloric acid-ethyl acetate solution (5 mL) and stirred at room temperature for 12 hours. After completion of the reaction, the solvent was distilled off under reduced pressure. The obtained residue was dissolved in acetonitrile (6 mL), iodotrimethylsilane (0.4 mL) was added under ice cooling, and the mixture was stirred at the same temperature for 2 hr. Water was added to the reaction solution, and the resulting solid was washed with water and acetonitrile in that order, and then dried at 80 ° C. under reduced pressure to obtain the desired product (280 mg) as a colorless solid.
¹ H NMR (DMSO-d ₆ , 400 MHz): δ0.84 (3H, t, J = 7.3 Hz), 1.27-1.29 (2H, m), 1.50-1.57 (2H, m), 1.71-1.73 (2H , m), 2.62 (3H, s), 2.69-2.71 (2H, m), 3.73-3.77 (2H, m), 6.98 (1H, s), 7.37 (1H, dd, J = 8.0, 1.8 Hz), 7.48 (1H, d, J = 8.0 Hz), 7.54 (1H, d, J = 1.8 Hz), 7.60 (1H, d, J = 8.6 Hz), 8.16 (1H, d, J = 8.6 Hz).
HRESIMS (+): 540.09836 (calculated as C ₂₂ H ₂₇ ClF ₃ NO ₅ PS 540.09882).

<Reference Example 19>
(S) -2-t-butoxycarbonylamino-4- {2-chloro-4- (2-methoxycarbonyl-5-trifluoromethylphenylthio) phenyl} -2-propylbutane-1-al

The compound of Reference Example 15 (704 mg) was dissolved in dimethyl sulfoxide (6 mL), triethylamine (1.7 mL) and sulfur trioxide-pyridine complex (973 mg) were added, and the mixture was stirred at room temperature for 1.5 hours. Ice water was added to the reaction solution, followed by extraction with ethyl acetate. The organic layer was washed with water and saturated brine, and then dried over anhydrous sodium sulfate. After evaporating the solvent under reduced pressure, the residue was purified by silica gel column chromatography (hexane: ethyl acetate = 5: 1) to obtain the desired product (595 mg) as a yellow oil.
¹ H NMR (CDCl ₃ , 400 MHz): δ 0.92 (3H, t, J = 7.3 Hz), 1.05-1.15 (1H, m), 1.26-1.35 (1H, m), 1.47 (9H, s), 1.63 -1.70 (1H, m), 2.02-2.07 (1H, m), 2.17-2.21 (1H, m), 2.52-2.58 (2H, m), 2.69-2.72 (2H, m), 3.98 (3H, s) , 5.41 (1H, brs), 7.06 (1H, s), 7.26 (1H, d, J = 8.0 Hz), 7.34-7.39 (2H, m), 7.53 (1H, d, J = 1.8 Hz), 8.07 ( 1H, d, J = 8.0 Hz), 9.32 (1H, s).
ESIMS (+): 574 [M + H] ⁺ .

<Reference Example 20>
(S) -3-t-Butoxycarbonylamino-5- {2-chloro-4- (2-methoxycarbonyl-5-trifluoromethylphenylthio) phenyl} -3-propyl-1-pentenylphosphonic acid dimethyl ester

Under argon gas atmosphere, tetramethyl methylenediphosphonate (242 mg) was dissolved in tetrahydrofuran (9 mL), and n-butyllithium (1.65 mol / L hexane solution, 0.63 mL) was added dropwise at -78 ° C. The mixture was stirred at the same temperature for 30 minutes. A tetrahydrofuran solution (3 mL) of the compound of Reference Example 19 (498 mg) was added dropwise to the reaction solution at −78 ° C., and the mixture was stirred at room temperature for 2 hours. A saturated aqueous ammonium chloride solution was added to the reaction mixture, and the mixture was extracted with ethyl acetate. The organic layer was washed with saturated brine and dried over anhydrous sodium sulfate. After evaporating the solvent under reduced pressure, the residue was purified by silica gel column chromatography (hexane: ethyl acetate = 3: 1 → ethyl acetate alone) to obtain the desired product (370 mg) as a colorless amorphous product.
¹ H NMR (CDCl ₃ , 400 MHz): δ 0.94 (3H, t, J = 7.3 Hz), 1.24-1.34 (2H, m), 1.45 (9H, s), 1.24-1.34 (2H, m), 1.61 -1.76 (2H, m), 2.04-2.21 (1H, m), 2.66-2.77 (1H, m), 3.73 (3H, s), 3.76 (3H, s), 3.98 (3H, s), 4.64 (1H , brs), 5.70 (1H, t, J = 17.7 Hz), 6.76 (1H, t, J = 17.7 Hz), 7.06 (1H, s), 7.28 (1H, d, J = 8.0 Hz), 7.34-7.39 (2H, m), 7.54 (1H, d, J = 1.8 Hz), 8.07 (1H, d, J = 8.0 Hz).
ESIMS (+): 680 [M + H] ⁺ .

<Reference Example 21>
(S) -3-t-Butoxycarbonylamino-5- {2-chloro-4- (2-methoxycarbonyl-5-trifluoromethylphenylthio) phenyl} -3-propylpentylphosphonic acid dimethyl ester

The compound of Reference Example 20 (369 mg) was dissolved in pyridine (11 mL), dipotassium azodicarboxylate (1.06 g) and acetic acid (0.47 mL) were added, and the mixture was stirred at room temperature for 64 hours. The reaction solution was diluted with toluene, and insoluble materials were removed using celite. After evaporating the solvent under reduced pressure, the residue was purified by silica gel column chromatography (hexane: ethyl acetate = 1: 2) to obtain the desired product (177 mg) as a colorless amorphous product.
¹ H NMR (CDCl ₃ , 400 MHz): δ 0.96 (3H, t, J = 7.3 Hz), 1.32-1.37 (2H, m), 1.45 (9H, s), 1.51-1.56 (2H, m), 1.71 -1.90 (4H, m), 2.05-2.14 (2H, m), 2.69-2.73 (2H, m), 3.74 (3H, s), 3.77 (3H, s), 3.98 (3H, s), 4.30 (1H , brs), 7.06 (1H, s), 7.29 (1H, d, J = 8.0 Hz), 7.35-7.39 (2H, m), 7.54 (1H, d, J = 1.8 Hz), 8.08 (1H, d, J = 8.0 Hz).
ESIMS (+): 682 [M + H] ⁺ .

 (S) -3-Amino-5- {4- (2-carboxy-5-trifluoromethylphenylthio) -2-chlorophenyl} -3-propylpentylphosphonic acid

Using the compound of Reference Example 21 (177 mg), the reaction was carried out in the same manner as in Example 1 to obtain the desired product (75 mg) as a colorless solid.
¹ H NMR (DMSO-d ₆ , 400 MHz): δ 0.90 (3H, t, J = 7.3 Hz), 1.33 (2H, brs), 1.61 (4H, brs), 1.83-1.90 (4H, m), 2.74 (2H, brs), 6.82 (1H, s), 7.27 (1H, d, J = 6.7 Hz), 7.43-7.46 (3H, m), 8.00 (1H, d, J = 8.0 Hz).
HRESIMS (+): 540.09808 (calculated as C ₂₂ H ₂₇ ClF ₃ NO ₅ PS 540.09882).

<Reference Example 22>
(S) -2-t-Butoxycarbonylamino-4- [2-chloro-4- (2-methoxy-5-trifluoromethylphenylthio) phenyl] -2-propylbutan-1-ol

The compound of Reference Example 14 (11.0 g) and the compound of Reference Example 3 (18.5 g) were reacted in the same manner as in Reference Example 15 to obtain the desired product (19.2 g) as a colorless oil.
¹ H NMR (CDCl ₃ , 400 MHz): δ 0.96 (3H, t, J = 7.3 Hz), 1.29-1.43 (2H, m), 1.44 (9H, s), 1.59 (2H, dd, J = 7.3, 1.8 Hz), 1.83 (1H, ddd.J = 13.9, 12.1, 5.4 Hz), 1.96 (1H, ddd, J = 13.9, 12.1, 5.4 Hz), 2.63-2.78 (2H, m), 3.75 (2H, d , J = 6.7 Hz), 3.92 (3H, s), 4.17 (1H, br s), 4.64 (1H, s), 6.96 (1H, d, J = 8.5 Hz), 7.17 (1H, dd, J = 8.5 , 1.8 Hz), 7.17 (1H, d, J = 8.5 Hz), 7.32-7.35 (2H, m), 7.51 (1H, dd, J = 8.5, 1.8 Hz).
ESIMS (+): 548 [M + H] ⁺ .

<Reference Example 23>
(S) -2-t-butoxycarbonylamino-4- [2-chloro-4- (2-hydroxy-5-trifluoromethylphenylthio) phenyl] -2-propylbutan-1-ol

Under argon atmosphere and ice-cooling, boron tribromide-methylene chloride solution (1 mol / L, 26 mL) was added to a solution of the compound of Reference Example 22 (3.69 g) in methylene chloride (43 mL) and ice-cooled. Stirred under for 1 hour. A saturated aqueous sodium hydrogen carbonate solution was added to the reaction mixture, and the mixture was extracted with ethyl acetate, washed with water and saturated brine in that order, and dried over anhydrous sodium sulfate. The solvent was distilled off under reduced pressure, the residue was dissolved in acetonitrile (43 mL) solvent, di-tert-butoxy dicarbonate (2.78 g) was added, and the mixture was stirred at room temperature for 2 hours, and then left overnight. Water was added to the reaction mixture, and the mixture was extracted with ethyl acetate, washed successively with water and saturated brine, and dried over anhydrous sodium sulfate. The solvent was distilled off under reduced pressure, and the residue was purified by silica gel column chromatography (hexane: ethyl acetate = 2: 1) to obtain the desired product (3.34 g) as a colorless oil.
¹ H NMR (CDCl ₃ , 400 MHz): δ 0.95 (3H, t, J = 7.3 Hz), 1.28-1.41 (2H, m), 1.44 (9H, s), 1.59 (2H, dd, J = 7.3, 1.8 Hz), 1.79 (1H, ddd.J = 13.9, 11.6, 5.5 Hz), 1.90 (1H, ddd, J = 13.9, 11.6, 5.5 Hz), 2.57-2.72 (2H, m), 3.72 (2H, d , J = 6.7 Hz), 4.16 (1H, br s), 4.61 (1H, br s), 6.77 (1H, s), 6.91 (1H, dd, J = 7.9, 1.8 Hz), 7.09 (1H, d, J = 1.8 Hz), 7.13 (1H, d, J = 7.9 Hz), 7.16 (1H, J = 7.9 Hz), 7.63 (1H, dd, J = 7.9, 1.8 Hz), 7.80 (1H, d, J = 1.8 Hz).
ESIMS (+): 534 [M + H] ⁺ .

<Reference Example 24>
(S) -2-t-butoxycarbonylamino-4- [2-chloro-4- (2-ethoxy-5-trifluoromethylphenylthio) phenyl] -2-propylbutan-1-ol

The compound of Reference Example 23 (240 mg) was dissolved in N, N-dimethylformamide (4.0 mL), and tetrabutylammonium iodide (33 mg), potassium carbonate (68.4 mg) and ethyl iodide (39.6 mg) were added. The mixture was stirred at room temperature for 15 hours. Water was added to the reaction mixture, and the mixture was extracted with ethyl acetate. The organic layer was washed with water and saturated brine, and then dried over anhydrous sodium sulfate. After evaporating the solvent under reduced pressure, the residue was purified by silica gel column chromatography (hexane: ethyl acetate = 2: 1) to obtain the desired product (243 mg) as a colorless oil.
¹ H NMR (CDCl ₃ , 400 MHz): δ 0.96 (3H, t, J = 7.3 Hz), 1.35-1.42 (2H, m), 1.37 (3H, t, J = 7.3 Hz), 1.44 (9H, s ), 1.54-1.60 (2H, m), 1.82 (1H, ddd, J = 13.9, 12.1, 5.4 Hz), 1.95 (1H, ddd, J = 13.9, 12.1, 5.4 Hz), 2.63-2.79 (2H, m ), 3.74 (2H, d, J = 6.7 Hz), 4.11 (2H, q, J = 7.3 Hz), 4.17 (1H, br s), 4.64 (1H, s), 6.91 (1H, d, J = 8.5 Hz), 7.19 (2H, s), 7.35 (1H, d, J = 1.8 Hz), 7.36 (1H, s), 7.47 (1H, dd, J = 8.5, 1.8 Hz).
ESIMS (+): 562 [M + H] ⁺ .

<Reference Example 25>
(S) -2-t-butoxycarbonylamino-4- [2-chloro-4- (2-methoxy-5-trifluoromethylphenylthio) phenyl] -1-dimethoxyphosphoryloxy-2-propylbutane

Using the compound of Reference Example 22 (194 mg), the reaction was carried out in the same manner as in Reference Example 16 to obtain the desired product (180 mg) as a colorless solid.
¹ H NMR (CDCl ₃ , 400 MHz): δ 0.96 (3H, t, J = 7.3 Hz), 1.31-1.42 (2H, m), 1.44 (9H, s), 1.62-1.75 (1H, m), 1.76 -1.89 (1H, m), 1.95-2.13 (2H, m), 3.74 (2H, t, J = 8.6 Hz), 3.78 (3H, d, J = 11.5 Hz), 3.79 (3H, d, J = 11.5 Hz), 3.92 (3H, s), 4.12 (1H, dd, J = 9.7, 4.8 Hz), 4.25 (1H, dd, J = 9.7, 4.8 Hz), 4.52 (1H, br s), 6.96 (1H, d, J = 8.5 Hz), 7.16 (1H, dd, J = 8.5, 1.8 Hz), 7.19 (1H, d, J = 8.5 Hz), 7.33 (2H, dd, J = 8.5, 1.8 Hz), 7.35 ( 1H, d, J = 1.8 Hz), 7.51 (1H, dd, J = 8.5, 1.8 Hz).
ESIMS (+): 656 [M + H] ⁺ .

<Reference Example 26>
(S) -2-t-butoxycarbonylamino-4- [2-chloro-4- (2-ethoxy-5-trifluoromethylphenylthio) phenyl] -1-dimethoxyphosphoryloxy-2-propylbutane

Using the compound of Reference Example 24 (200 mg), the reaction was carried out in the same manner as in Reference Example 16 to obtain the desired product (180 mg) as a colorless solid.
¹ H NMR (CDCl ₃ , 400 MHz): δ 0.96 (3H, t, J = 7.3 Hz), 1.35-1.42 (2H, m), 1.37 (3H, t, J = 7.3 Hz), 1.44 (9H, s ), 1.59-1.74 (2H, m), 1.75-1.86 (1H, m), 1.96-2.12 (1H, m), 2.66-2.75 (2H, m), 3.78 (3H, d, J = 10.9 Hz), 3.79 (3H, d, J = 10.9 Hz), 4.11 (2H, q, J = 7.3 Hz), 4.12 (1H, dd, J = 10.3, 4.8 Hz), 4.25 (1H, dd, J = 10.3, 4.8 Hz) ), 4.52 (1H, br s), 6.91 (1H, d, J = 8.5 Hz), 7.18 (2H, s), 7.35 (1H, dd, J = 8.5, 1.8 Hz), 7.36 (1H, s), 7.47 (1H, dd, J = 8.5, 1.8 Hz).
ESIMS (+): 670 [M + H] ⁺ .

<Reference Example 27>
(S) -2-Amino-4- [2-chloro-4- (2-methoxy-5-trifluoromethylphenylthio) phenyl] -2-propylbutyl phosphate monoester

Using the compound of Reference Example 25 (180 mg), the reaction was carried out in the same manner as in Example 1 to obtain the desired product (113 mg) as a colorless solid.
¹ H NMR (DMSO-d ₆ -dTFA, 400 MHz): δ 0.89 (3H, t, J = 7.3 Hz), 1.24-1.39 (2H, m), 1.45-1.81 (4H, m), 2,59- 2.75 (2H, m), 3.70-3.83 (2H, m), 3.89 (3H, s), 7.25 (1H, dd, J = 8.5, 1.8 Hz), 7.27 (1H, d, J = 1.8 Hz), 7.28 (1H, d, J = 8.5 Hz), 7.37 (1H, d, J = 1.8 Hz), 7.38 (1H, d, J = 8.5 Hz), 7.69 (1H, dd, J = 8.6, 1.8 Hz), 8.73 (2H, br s).
HRESIMS (+): 518.09956 (Calculated value as C ₂₁ H ₂₇ ClF ₃ NO ₅ PS 518.09882).

<Reference Example 28>
(S) -2-Amino-4- [2-chloro-4- (2-ethoxy-5-trifluoromethylphenylthio) phenyl] -2-propylbutyl phosphate monoester

Using the compound of Reference Example 26 (180 mg), the reaction was carried out in the same manner as in Example 1 to obtain the desired product (110 mg) as a colorless solid.
¹ H NMR (DMSO-d ₆ -dTFA, 400 MHz): δ 0.89 (3H, t, J = 7.3 Hz), 1.23 (3H, t, J = 7.3 Hz), 1.25-1.39 (2H, m), 1.44 -1.83 (4H, m), 2,60-2.76 (2H, m), 3.70-3.83 (2H, m), 4.14 (2H, q, J = 7.3 Hz), 7.24 (1H, d, J = 8.5 Hz ), 7.26 (1H, dd, J = 8.5, 1.8 Hz), 7.31 (1H, d, J = 1.8 Hz), 7.38 (1H, d, J = 8.5 Hz), 7.39 (1H, d, J = 1.8 Hz) ), 7.65 (1H, dd, J = 8.6, 1.8 Hz), 8.67 (2H, br s).
HRESIMS (+): 542.11446 (Calculated value as C ₂₂ H ₂₉ ClF ₃ NO ₅ PS 542.11447).

Next, with respect to the compounds exemplified as Examples, the results supporting the usefulness are shown by Experimental Example 1, Experimental Example 2, and Experimental Example 3.

<Experimental example 1> S1P (sphingosine-1-phosphate) suppresses the effect of a test compound on intracellular calcium mobilization in human S1P3 receptor-expressing cells

Human S1P3 receptor-expressing CHO cells were subcultured in Ham's F-12 medium containing 10% fetal bovine serum and 300 μg / mL Geneticin. The human S1P3 receptor-expressing CHO cells were treated with 0.25% trypsin, recovered from the dish, and suspended in Ham's F-12 medium containing 10% fetal bovine serum and 300 μg / mL Geneticin. Then, were seeded in 96-well black clear bottom plates at the human S1P3 receptor expressing CHO cells is ^{2.5x 10 4/100 μL / well} (BD Falcon Biocoat), 37 ℃, two nights cultured under 5% CO ₂ did. The next day, the wells were washed three times with Ham's F-12 medium containing 100 μL 0.1% fatty acid-free bovine serum albumin (BSA). After exchanging with Ham's F-12 medium containing 0.1% BSA, serum starvation treatment was performed in a 37 ° C. CO ₂ incubator for 6 hours.

After 6 hours, the medium was discarded, 50 μL / well of Fluo3 loading buffer was added, and the cells were further cultured for 1 hour. The Fluo3 loading buffer was prepared as follows. First, Fluo3-AM (Dojindo) and pluronic F-127 (20% DMSO solution, invitrogen) were mixed in equal amounts. Subsequently, the mixed solution of Fluo3-AM and pluronic F-127 was added to Hanks-HEPES buffer (20 mM HEPES (pH 7.4), 0.1% BSA (Fatty acid Free), 2.5 mM probenecid-containing Hanks balanced salt solution). In addition, the Fluo3-AM final concentration of 4 μM was used as the Fluo3 loading buffer.

After 1 hour incubation, it was washed 3 times with 100 μL Hanks-HEPES buffer. Add 100 μL of the test compound (0.125 nM, 1.25 nM, 12.5 nM, 125 nM, 1.25 μM) or the same buffer in which DMSO is dissolved, and in a microplate fluorescence spectrophotometer (FLEX Station (Molecular Device)) at 37 ° C. Incubated for 30 minutes. Then, using the same device, add 25 μL of S1P (final concentrations 0.1 nM, 1 nM, 10 nM, 100 nM, 1 μM) prepared at a concentration 5 times the final concentration by serial dilution method, and based on calcium mobilization Fluo3 fluorescence was detected and measured at an excitation wavelength of 485 nm and a detection wavelength of 525 nm. Based on the measurement data, a value (fluorescence increase amount) obtained by subtracting the minimum fluorescence intensity from the maximum fluorescence intensity was calculated. Using the measured fluorescence increase amount, PRISM 4 software (GraphPad) was used to approximate the relationship between the concentration of S1P and the fluorescence increase curve. Based on the results, EC50 values were calculated when the compound was not treated and when each concentration of compound was treated. Based on these values, Schild Plot analysis was performed to determine the dissociation constant Kd value. In addition, 1000 nmol / L> Kd value ≧ 100 nmol / L +, 100 nmol / L> Kd value ≧ 10 nmol / L ++, 10 nmol / L> Kd value ≧ 1 nmol / L + ++, 1 nmol / L> Kd values are expressed as ++++ and shown in Table 1.

＜実験例２＞ ヒトS1P1受容体発現細胞に対する被験化合物の細胞内カルシウム動員誘導試験

<Experimental example 2> Intracellular calcium mobilization induction test of test compound against human S1P1 receptor expressing cells

Human S1P1 receptor-expressing CHO cells were subcultured in Ham's F-12 medium containing 10% fetal bovine serum and 300 μg / mL Geneticin. The human S1P1 receptor-expressing CHO cells were treated with 0.25% trypsin, recovered from the dish, and suspended in Ham's F-12 medium containing 10% fetal bovine serum and 300 μg / mL Geneticin. Then, were seeded in 96-well black clear bottom plates at the human S1P1 receptor expressing CHO cells is ^{2.5 x 10 4/100 μL /} well (BD Falcon Biocoat), 37 ℃, two nights cultured under 5% CO ₂ did. The next day, the wells were washed three times with Ham's F-12 medium containing 100 μL 0.1% fatty acid-free bovine serum albumin (BSA). After exchanging with Ham's F-12 medium containing 0.1% BSA, serum starvation treatment was performed in a 37 ° C. CO ₂ incubator for 6 hours.

After 6 hours, the medium was discarded, 50 μL / well of Fluo3 loading buffer was added, and the cells were further cultured for 1 hour. The Fluo3 loading buffer was prepared as follows. First, Fluo3-AM (Dojindo) and pluronic F-127 (20% DMSO solution, invitrogen) were mixed in equal amounts. Subsequently, the mixed solution of Fluo3-AM and pluronic F-127 was added to Hanks-HEPES buffer (20 mM HEPES (pH 7.4), 0.1% BSA (Fatty acid Free), 2.5 mM probenecid-containing Hanks balanced salt solution). In addition, the Fluo3-AM final concentration of 4 μM was used as the Fluo3 loading buffer.

After 1 hour incubation, it was washed 3 times with 100 μL Hanks-HEPES buffer. Next, 100 μL of the same buffer was added and incubated at 37 ° C. for 15 minutes in a microplate fluorescence spectrophotometer (FLEX Station (Molecular Device)). After that, using the same device, S1P or test compound (final concentration 0.1 nM, 1 nM, 10 nM, 100 nM, 1 μM) prepared in the same buffer in which DMSO was dissolved or serial dilution method was used. , 10 μM) was added, and fluorescence by Fluo3 based on calcium mobilization was detected and measured at an excitation wavelength of 485 nm and a detection wavelength of 525 nm. Based on the measurement data, calculate the value obtained by subtracting the minimum fluorescence intensity from the maximum fluorescence intensity (fluorescence increase), and increase the fluorescence when the solvent is added and increase the fluorescence when S1P is applied at 10 ^-6 M The difference in fluorescence was taken as 100% to calculate the fluorescence increase rate (%) of the test compound. The EC ₅₀ value was determined using PRISM software (GraphPad) as the intracellular calcium mobilization inducing action of the test compound.

The EC ₅₀ values of the compounds of Example 1 and Example 2 were> 10 μmol / L. Moreover, as a result of evaluating the antagonistic action of the S1P1 receptor using the method of Experimental Example 1, the Kd value of the compounds of Example 1 and Example 2 was> 1.0 μmol / L.

<Experimental example 3> Sepsis model with cecal perforation

  This model is widely used as a polymicrobial abdominal sepsis model with intestinal bacterial leakage. The method described in Non-Patent Document 9 (D. Rittirsch et al., Nature Protocols, 4, 31 (2009)) was used as a reference.

Wistar Rat rats (Nippon Charles River male 8W) were used. Under anesthesia with isoflurane, the abdomen of the rat was incised and the cecum was exposed. The cecum was tied with sterilized silk thread, and 10 holes were made in the cecal tip using an 18G needle. After the treatment, the cecum was returned to the body, and the wound was sutured. Furthermore, physiological saline was administered subcutaneously at 30 mL / kg. Rats were then returned to their cages and observed for 3 days to determine survival. The test compound was continuously administered from 1 hour after the CLP treatment so as to be 0.1 mg / kg / hr from a cannula placed in the femoral vein.

In the administration group of the compound of Example 1, a statistically significant action (survival extension action, Log-rank test p <0.05) that shifts the survival curve to the right as compared with the vehicle administration group was observed. In the vehicle administration group, all cases died before the lapse of 1 day, whereas in the administration group of Example 1, 40% survived after 1 day and 12.5% survived after 2 days. The survival rate was improved. This result suggests that the compound of Example 1 is effective against sepsis.

From the above results, it has been clarified that the compound of the present invention exhibits an excellent antagonistic action on the human S1P3 receptor, but has weak or no antagonistic action or agonistic action on the S1P1 receptor. It was also confirmed that the compound of the present invention has an excellent inhibitory effect on sepsis.

  The present invention can provide a diphenyl sulfide derivative having excellent S1P3 antagonist activity and S1P3 selectivity. Further, the diphenyl sulfide derivative of the present invention can be used safely as a medicine because it has weak or no hemolytic, tissue damage and central inhibitory action. Furthermore, the diphenyl sulfide derivative of the present invention is stable in an aqueous solution. The compound of the present invention having these excellent properties is used for bronchial constriction, bronchial asthma, chronic obstructive pulmonary disease (COPD), pulmonary emphysema, tracheal stenosis, diffuse panbronchiolitis, infection, connective tissue disease or bronchi associated with transplantation. Inflammation, diffuse hamartoma pulmonary vascular myomatosis, adult respiratory distress syndrome (ARDS), interstitial pneumonia, lung cancer, hypersensitivity pneumonitis, idiopathic interstitial pneumonia, pulmonary fibrosis, sepsis, influenza virus, RS Caused by cytokine storm based on viral infection, arteriosclerosis, intimal thickening, solid tumor, diabetic retinopathy, rheumatoid arthritis, heart failure, ischemic reperfusion injury, cerebral vascular spasm after subarachnoid hemorrhage, coronary vascular spasm Lung disease caused by pulmonary edema such as angina pectoris or myocardial infarction, glomerulonephritis, thrombosis, ARDS, cardiac arrhythmia, eye disease, ocular hypertension, glaucoma, glaucomatous retinopathy, Neurosis, it is useful as a preventive or therapeutic agent for macular degeneration.

Claims (8)

General formula (1)

[In Formula (1), R ¹ represents an alkyl group having 1 to 6 carbon atoms or a lower alkoxy group having 1 to 6 carbon atoms which may be substituted with ¹ to 3 halogen atoms, and R ² represents the number of carbon atoms. An alkyl group having 1 to 6 carbon atoms or an alkenyl group having 2 to 6 carbon atoms; R ¹⁰ represents an acyl group having 1 to 6 carbon atoms or a carboxyl group; X represents a methylene or oxygen atom; and Z represents a halogen atom. ]
Or a pharmacologically acceptable salt thereof or a hydrate thereof. The compound represented by the general formula (1) is represented by the general formula (1a).

[In formula (1a), R ² and R ¹⁰ are as described above]
The diphenyl sulfide derivative of Claim 1 represented by these, its pharmacologically acceptable salt, or those hydrates. The compound represented by the general formula (1) is
(S) -2-amino-4- {4- (2-carboxy-5-trifluoromethylphenylthio) -2-chlorophenyl} -2-propylbutyl phosphate monoester,
(S) -4- [4- (2-acetyl-5-trifluoromethylphenylthio) -2-chlorophenyl] -2-amino-2-propylbutyl phosphate monoester, or
The diphenyl sulfide derivative or drug according to claim 1, which is (S) -3-amino-5- {4- (2-carboxy-5-trifluoromethylphenylthio) -2-chlorophenyl} -3-propylpentylphosphonic acid. Physiologically acceptable salts or hydrates thereof. A sphingosine-1-phosphate 3 (S1P3) receptor antagonist comprising the diphenyl sulfide derivative according to any one of claims 1 to 3 or a pharmacologically acceptable salt thereof or a hydrate thereof as an active ingredient. Drug based on action. Airway constriction, bronchial asthma, chronic obstructive pulmonary disease (COPD), emphysema, tracheal stenosis, diffuse panbronchiolitis, bronchitis associated with infection, connective tissue disease or transplantation, diffuse hamartoma pulmonary vascular myomatosis , Adult respiratory distress syndrome (ARDS), interstitial pneumonia, lung cancer, hypersensitivity pneumonitis, idiopathic interstitial pneumonia, pulmonary fibrosis, sepsis, or cytokine storm based on influenza virus or RS virus infection The medicament according to claim 4, wherein Arteriosclerosis, intimal thickening, solid tumor, diabetic retinopathy, rheumatoid arthritis, heart failure, ischemic reperfusion injury, cerebrovascular spasm after subarachnoid hemorrhage, angina or myocardial infarction caused by coronary vascular spasm Claim 1, which is a therapeutic or preventive agent for glomerulonephritis, thrombosis, lung disease caused by pulmonary edema, cardiac arrhythmia, eye disease, ocular hypertension, glaucoma, glaucomatous retinopathy, optic neuropathy or macular degeneration 4. The medicine according to 4. The medicament according to claim 4, which is a therapeutic or prophylactic agent for sepsis. A pharmaceutical composition comprising the diphenyl sulfide derivative according to any one of claims 1 to 3 or a pharmacologically acceptable salt thereof or a hydrate thereof and a pharmaceutically acceptable carrier.