Patent Application
20090131438
This invention relates to novel triazole derivatives having the effect of inhibiting binding between sphingosine-1-phosphate having various physiological actions and Edg-1 (Endothelial differentiation gene receptor type-1, S1P1), which is one of its receptors, and pharmaceutical products containing them as active ingredients.
Sphingosine-1-phosphate (hereinafter referred to as “S1P”) is a physiologically active lipid which is produced upon metabolism, in cells, of sphingolipids typified by sphingomyelin. S1P is known to have wide varieties of actions, including a cell differentiation inducing action, a cell growth promoting action, control of cell motility, and an antiapoptotic action, and to show physiological actions such as angiogenesis, induction of bradycardia, activation of inflammatory cells, and activation of platelets (non-patent document 1).
Five subtypes, Edg-1(S1P1), Edg-3(S1P3), Edg-5(S1P2), Edg-6(S1P4), and Edg-8(S1P5), are reported as receptors of S1P (non-patent document 2).
One of them, Edg-1(S1P1), is expressed in large amounts in immune cells such as T cells and dendritic cells, and the vascular endothelial cells, and is suggested to contribute deeply to S1P-associated migration of T cells (non-patent document 3), migration of mast cells (non-patent document 4), emigration of T cells and B cells from lymph organs (non-patent document 5), and angiogenesis (non-patent document 6), and to be involved in autoimmune diseases such as Crohn disease, irritable colitis, Sjögren syndrome, multiple sclerosis, and systemic lupus erythematosus, and diseases such as rheumatoid arthritis, asthma, atopic dermatitis, rejection reaction after organ transplantation, cancer, retinopathy, psoriasis, osteoarthrosis, and age-related macular degeneration.
Thus, Edg-1(S1P1) ligands are considered to be effective for the treatment or prevention of these diseases.
So far, certain types of thiophene derivatives (non-patent document 7), phosphate ester derivatives (patent document 1, patent document 2, non-patent document 8), and thiazolidine derivatives (patent document 3) have been reported as Edg-1(S1P1) ligands. However, inhibitors having structures similar to the structure of the compound of the present invention have not been known.
Compounds similar in structure to the compound of the present invention are marketed as reagents by Bionet, but their pharmaceutical uses are completely unknown.
It is an object of the present invention to provide a compound which has the effect of inhibiting binding between S1P and its receptor Edg-1(S1P1) and is useful as a pharmaceutical product.
The inventors diligently conducted studies in an attempt to find ligand compounds for Edg-1(S1P1). As a result, they have found that certain types of triazole derivatives or their pharmaceutically acceptable salts attain this object. This finding has led to the accomplishment of the present invention.
That is, the present invention is a compound represented by the formula (I) below or a pharmaceutically acceptable salt thereof
A represents a sulfur atom, an oxygen atom, a group represented by the formula —SO—, or a group represented by the formula —SO2—,
R1 represents a hydrogen atom, an alkyl group having 1 to 16 carbon atoms, an alkenyl group having 2 to 8 carbon atoms, an alkynyl group having 2 to 8 carbon atoms, [an alkyl group having 1 to 6 carbon atoms which has been substituted by a phenyl group, “a phenyl group substituted by 1 to 5 groups selected from a phenyl group, a cyano group, a halogen atom, an alkyl group having 1 to 6 carbon atoms, a trifluoromethyl group, a methoxycarbonyl group, an alkylthio group having 1 to 6 carbon atoms, a dimethylamino group, a nitro group, and an acetamido group”, a cycloalkyl group having 3 to 8 carbon atoms, a hydroxyl group, an alkylthio group having 1 to 6 carbon atoms, an alkoxy group having 1 to 6 carbon atoms, a benzyloxy group, a phenoxy group, a trifluoromethyl group, a difluoromethyl group, a benzenesulfonyl group, a naphthyl group, a tricycloalkyl group having 7 to 10 carbon atoms, a carbomethoxy(phenyl)methyl group, a diphenylmethyl group, a 1-phenylethyl group, an imidazolyl group, an indolyl group, a pyridyl group, an oxetanyl group, an oxoranyl group, a methylpiperidinyl group, a benzylpiperidinyl group, a morpholino group, a 2-oxopyrrolidin-1-yl group, a 2-oxoimidazolidin-1-yl group, a group represented by the formula
—CO2R11
—COR16
R2 represents an alkyl group having 1 to 6 carbon atoms, a cycloalkyl group having 3 to 8 carbon atoms, a phenyl group, or [an alkyl group having 1 to 6 carbon atoms which has been substituted by a phenyl group, an alkoxy group having 1 to 6 carbon atoms, a morpholino group, a piperidino group, a group represented by the formula
R3 represents a hydrogen atom or an alkyl group having 1 to 6 carbon atoms,
R4 represents a hydrogen atom, an alkyl group having 1 to 6 carbon atoms, a benzyl group, a substituted benzyl group, a phenethyl group, “an alkyl group having 1 to 6 carbon atoms which has been substituted by an alkoxy group having 1 to 6 carbon atoms, a halogen atom or a hydroxyl group”, or a phenyl group, or
R3 and R4 together form a 3- to 6-membered saturated hydrocarbon ring,
R5 represents a hydrogen atom or an alkyl group having 1 to 6 carbon atoms, and
Y represents a group represented by the formula
where R6 represents an alkyl group having 1 to 10 carbon atoms, an alkenyl group having 2 to 8 carbon atoms, “an alkyl group having 1 to 10 carbon atoms which has been substituted by 1 to 5 groups selected from a phenyl group, a substituted phenyl group, a cycloalkyl group having 3 to 8 carbon atoms, a halogen atom, a naphthyl group, a heterocyclic group, and a substituted heterocyclic group”, “an alkenyl group having 2 to 8 carbon atoms which has been substituted by 1 to 5 groups selected from a phenyl group, a substituted phenyl group, a cycloalkyl group having 3 to 8 carbon atoms, a halogen atom, a naphthyl group, a heterocyclic group, and a substituted heterocyclic group”, a phenyl group, a substituted phenyl group, a naphthyl group, a naphthyl group substituted by a dimethylamino group, a heterocyclic group, or a substituted heterocyclic group,
with the exception of [the compound in which A is an oxygen atom, and R1 is a hydrogen atom], [the compound in which A is a sulfur atom, and R1 is a hydrogen atom], [the compound in which A is a sulfur atom, and R3 and R4 are each a hydrogen atom at the same time], [the compound in which A is a sulfur atom, and R2 is a phenyl group], [the compound in which A is a sulfur atom, R2 is an ethyl group, R3 and R5 are each a hydrogen atom, R4 is a benzyl group, R6 is a 4-fluorophenyl group, and R1 is a 3-methoxybenzyl group], [the compound in which A is a sulfur atom, R2 is an ethyl group, R3 and R5 are each a hydrogen atom, R4 is a benzyl group, R6 is a 4-fluorophenyl group, and R1 is a 3-fluorobenzyl group], [the compound in which A is a sulfur atom, R2 is an ethyl group, R3 and R5 are each a hydrogen atom, R4 is a benzyl group, R6 is a 4-fluorophenyl group, and R1 is a 2-propenyl group], [the compound in which A is a sulfur atom, R2 is an ethyl group, R3 and R5 are each a hydrogen atom, R4 is a benzyl group, R6 is a 4-fluorophenyl group, and R1 is a 4-t-butylbenzyl group], [the compound in which A is a sulfur atom, R2 is an ethyl group, R3 and R5 are each a hydrogen atom, R4 is a benzyl group, R6 is a 4-fluorophenyl group, and R1 is a methyl group], [the compound in which A is a sulfur atom, R2 is an ethyl group, R3 and R5 are each a hydrogen atom, R4 is a methyl group, R6 is a 4-chlorophenyl group, and R1 is a methyl group], [the compound in which A is a sulfur atom, R2 is an ethyl group, R3 and R5 are each a hydrogen atom, R4 is a methyl group, R6 is a 4-chlorophenyl group, and R1 is a 2-propenyl group], [the compound in which A is a sulfur atom, R2 is an ethyl group, R3 and R5 are each a hydrogen atom, R4 is a methyl group, R6 is a 4-chlorophenyl group, and R1 is a 4-methoxybenzyl group], [the compound in which A is a sulfur atom, R2 is an ethyl group, R3 and R5 are each a hydrogen atom, R4 is a methyl group, R6 is a 4-chlorophenyl group, and R1 is a 4-t-butylbenzyl group], [the compound in which A is a sulfur atom, R2 is an ethyl group, R3 and R5 are each a hydrogen atom, R4 is a methyl group, R6 is a 4-chlorophenyl group, and R1 is a 3,4-dichlorobenzyl group], [the compound in which A is a sulfur atom, R2 is an ethyl group, R3 and R5 are each a hydrogen atom, R4 is a methyl group, R6 is a 4-chlorophenyl group, and R1 is a 2-chlorobenzyl group], [the compound in which A is a sulfur atom, R2 is an ethyl group, R3 and R5 are each a hydrogen atom, R4 is a methyl group, R6 is a 4-chlorophenyl group, and R1 is a 3-fluorobenzyl group], [the compound in which A is a sulfur atom, R2 is a methyl group, R3 and R5 are each a hydrogen atom, R4 is a benzyl group, R6 is a 4-chlorophenyl group, and R1 is a 2-methyl-2-propenyl group], [the compound in which A is a sulfur atom, R2 is a methyl group, R3 and R5 are each a hydrogen atom, R4 is a benzyl group, R6 is a 4-chlorophenyl group, and R1 is a 2-propenyl group], [the compound in which A is a sulfur atom, R2 is a methyl group, R3 and R5 are each a hydrogen atom, R4 is a benzyl group, R6 is a 4-chlorophenyl group, and R1 is a methoxycarbonylmethyl group], [the compound in which A is a sulfur atom, R2 is a methyl group, R3 and R5 are each a hydrogen atom, R4 is a benzyl group, R6 is a 4-chlorophenyl group, and R1 is a 4-trifluoromethylbenzyl group], [the compound in which A is a sulfur atom, R2 is a methyl group, R3 and R5 are each a hydrogen atom, R4 is a benzyl group, R6 is a 4-chlorophenyl group, and R1 is a 3,4-dichlorobenzyl group], [the compound in which A is a sulfur atom, R2 is a methyl group, R3 and R5 are each a hydrogen atom, R4 is a benzyl group, R6 is a 4-chlorophenyl group, and R1 is a 4-bromobenzyl group], and [the compound in which A is a sulfur atom, R2 is an ethyl group, R3 and R5 are each a hydrogen atom, R4 is a benzyl group, R6 is a 4-fluorophenyl group, and R1 is a 4-methylbenzyl group]}.
Another aspect of the present invention is a pharmaceutical product comprising a compound represented by the formula (I) below or a pharmaceutically acceptable salt thereof as an active ingredient:
A represents a sulfur atom, an oxygen atom, a group represented by the formula —SO—, or a group represented by the formula —SO2—,
R1 represents a hydrogen atom, an alkyl group having 1 to 16 carbon atoms, an alkenyl group having 2 to 8 carbon atoms, an alkynyl group having 2 to 8 carbon atoms, [an alkyl group having 1 to 6 carbon atoms which has been substituted by a phenyl group, “a phenyl group substituted by 1 to 5 groups selected from a phenyl group, a cyano group, a halogen atom, an alkyl group having 1 to 6 carbon atoms, a trifluoromethyl group, a methoxycarbonyl group, an alkylthio group having 1 to 6 carbon atoms, a dimethylamino group, a nitro group, and an acetamido group”, a cycloalkyl group having 3 to 8 carbon atoms, a hydroxyl group, an alkylthio group having 1 to 6 carbon atoms, an alkoxy group having 1 to 6 carbon atoms, a benzyloxy group, a phenoxy group, a trifluoromethyl group, a difluoromethyl group, a benzenesulfonyl group, a naphthyl group, a tricycloalkyl group having 7 to 10 carbon atoms, a carbomethoxy(phenyl)methyl group, a diphenylmethyl group, a 1-phenylethyl group, an imidazolyl group, an indolyl group, a pyridyl group, an oxetanyl group, an oxoranyl group, a methylpiperidinyl group, a benzylpiperidinyl group, a morpholino group, a 2-oxopyrrolidin-1-yl group, a 2-oxoimidazolidin-1-yl group, a group represented by the formula
—CO2R11
—COR16
R2 represents an alkyl group having 1 to 6 carbon atoms, a cycloalkyl group having 3 to 8 carbon atoms, a phenyl group, or [an alkyl group having 1 to 6 carbon atoms which has been substituted by a phenyl group, an alkoxy group having 1 to 6 carbon atoms, a morpholino group, a piperidino group, a group represented by the formula
R3 represents a hydrogen atom or an alkyl group having 1 to 6 carbon atoms,
R4 represents a hydrogen atom, an alkyl group having 1 to 6 carbon atoms, a benzyl group, a substituted benzyl group, a phenethyl group, “an alkyl group having 1 to 6 carbon atoms which has been substituted by an alkoxy group having 1 to 6 carbon atoms, a halogen atom or a hydroxyl group”, or a phenyl group, or
R3 and R4 together form a 3- to 6-membered saturated hydrocarbon ring,
R5 represents a hydrogen atom or an alkyl group having 1 to 6 carbon atoms, and
Y represents a group represented by the formula
where R6 represents an alkyl group having 1 to 10 carbon atoms, an alkenyl group having 2 to 8 carbon atoms, “an alkyl group having 1 to 10 carbon atoms which has been substituted by 1 to 5 groups selected from a phenyl group, a substituted phenyl group, a cycloalkyl group having 3 to 8 carbon atoms, a halogen atom, a naphthyl group, a heterocyclic group, and a substituted heterocyclic group”, “an alkenyl group having 2 to 8 carbon atoms which has been substituted by 1 to 5 groups selected from a phenyl group, a substituted phenyl group, a cycloalkyl group having 3 to 8 carbon atoms, a halogen atom, a naphthyl group, a heterocyclic group, and a substituted heterocyclic group”, a phenyl group, a substituted phenyl group, a naphthyl group, a naphthyl group substituted by a dimethylamino group, a heterocyclic group, or a substituted heterocyclic group}.
Still another aspect of the present invention is the compound of the formula (I) or the pharmaceutically acceptable salt thereof according to claim 1 wherein the portion corresponding to Y is a hydrogen atom, A is an oxygen atom, and R5 is a hydrogen atom, and is an intermediate useful for producing the compound of the formula (I).
In the present invention, the alkyl group having 1 to 16 carbon atoms refers to a straight-chain or branched-chain alkyl group having 1 to 16 carbon atoms. Its examples are a methyl group, an ethyl group, an n-propyl group, an isopropyl group, an n-butyl group, an isobutyl group, a tert-butyl group, a sec-butyl group, an n-pentyl group, an isopentyl group, a neopentyl group, a tert-pentyl group, an n-hexyl group, an n-heptyl, an n-octyl group, and an n-hexadecyl group.
The alkyl group having 1 to 6 carbon atoms refers to a straight-chain or branched-chain alkyl group having 1 to 6 carbon atoms. Its examples are a methyl group, an ethyl group, an n-propyl group, an isopropyl group, an n-butyl group, an isobutyl group, a tert-butyl group, a sec-butyl group, an n-pentyl group, an isopentyl group, a neopentyl group, a tert-pentyl group, and an n-hexyl group.
The alkyl group having 1 to 4 carbon atoms refers to a straight-chain or branched-chain alkyl group having 1 to 4 carbon atoms. Its examples are a methyl group, an ethyl group, an n-propyl group, an isopropyl group, an n-butyl group, an isobutyl group, a tert-butyl group, and a sec-butyl group.
The alkenyl group having 2 to 8 carbon atoms refers to a straight-chain or branched-chain alkenyl group having 2 to 8 carbon atoms. Its examples are a vinyl group, an allyl group, a 1-propenyl group, an isopropenyl group, a 1-butenyl group, a 2-butenyl group, a 3-butenyl group, a 1,3-butadienyl group, a 2-methylallyl group, a 2-methyl-propenyl group, a 2-pentenyl group, and a 3-methyl-but-2-enyl group.
The alkenyl group having 3 to 5 carbon atoms refers to a straight-chain or branched-chain alkenyl group having 3 to 5 carbon atoms. Its examples are an allyl group, a 1-propenyl group, an isopropenyl group, a 1-butenyl group, a 2-butenyl group, a 3-butenyl group, a 1,3-butadienyl group, a 2-methylallyl group, a 2-methyl-propenyl group, and a 4-pentenyl group.
The alkynyl group having 2 to 8 carbon atoms refers to a straight-chain or branched-chain alkynyl group having 2 to 8 carbon atoms. Its examples are an ethynyl group, a 2-propynyl group, a 2-butynyl group, a 1-methyl-prop-2-ynyl group, a 2-pentynyl group, and a 4-pentynyl group.
The cycloalkyl group having 3 to 8 carbon atoms refers to a cycloalkyl group having 3 to 8 carbon atoms, and its examples are a cyclopropyl group, a cyclobutyl group, a cyclopentyl group, a cyclohexyl group, a cycloheptyl group, and a cyclooctyl group.
The cycloalkyl group having 3 to 6 carbon atoms refers to a cycloalkyl group having 3 to 6 carbon atoms, and its examples are a cyclopropyl group, a cyclobutyl group, a cyclopentyl group, and a cyclohexyl group.
The halogen atom refers to a fluorine atom, a chlorine atom, a bromine atom, or an iodine atom.
The alkylthio group having 1 to 6 carbon atoms refers to a straight-chain or branched-chain alkylthio group having 1 to 6 carbon atoms. Its examples are a methylthio group, an ethylthio group, a propylthio group, an isopropylthio group, a butylthio group, an isobutylthio group, a pentylthio group, a hexylthio group, and an allylthio group.
The alkoxy group having 1 to 6 carbon atoms refers to a straight-chain or branched-chain alkoxy group having 1 to 6 carbon atoms. Its examples are a methoxy group, an ethoxy group, a propoxy group, an isopropoxy group, a butoxy group, an isobutoxy group, a sec-butoxy group, a tert-butoxy group, a pentyloxy group, a hexyloxy group, and an allyloxy group.
The cycloalkyl group having 3 to 8 carbon atoms refers to a cycloalkyl group having 3 to 8 carbon atoms, and its examples are a cyclopropyl group, a cyclobutyl group, a cyclopentyl group, a cyclohexyl group, a cycloheptyl group, and a cyclooctyl group.
The tricycloalkyl group having 7 to 10 carbon atoms refers to a tricycloalkyl group having 7 to 10 carbon atoms, and includes, for example, an adamantyl group.
The cycloalkyl group having 3 to 8 carbon atoms in which the benzene rings have been condensed includes, for example, a 1,2,3,4-tetrahydronaphthalenyl group, and an indanyl group.
The substituted benzyl group refers to a benzyl group substituted by (1 to 2 groups selected from a phenyl group, a halogen atom, a methyl group, a methoxy group, a trifluoromethyl group, or a hydroxyl group). Its examples are a 4-phenylbenzyl group, a 3,4-dichlorobenzyl group, a 4-methylbenzyl group, and a 4-methoxybenzyl group.
As the 3- to 6-membered saturated hydrocarbon ring, cyclopropane, cyclobutane, cyclopentane, and cyclohexane can be named.
The alkyl group having 1 to 10 carbon atoms refers to a straight-chain or branched-chain alkyl group having 1 to 10 carbon atoms. Its examples are a methyl group, an ethyl group, an n-propyl group, an isopropyl group, an n-butyl group, an isobutyl group, a tert-butyl group, a sec-butyl group, an n-pentyl group, an isopentyl group, a neopentyl group, a tert-pentyl group, an n-hexyl group, and an n-decyl group.
The substituted phenyl group refers to a phenyl group substituted by 1 to 5 groups selected, for example, from a phenyl group, a methoxy group, a phenyl group substituted by an acetyl group, an oxazolyl group, a pyrazolyl group, a methylpyrimidinyl group, a cyano group, a halogen atom, an alkyl group having 1 to 6 carbon atoms, a trifluoromethyl group, a hydroxyl group, an alkoxy group having 1 to 6 carbon atoms, a cyanoethoxy group, a phenoxy group, a phenoxy group substituted by a methoxy group, a pyridinyloxy group, an acetyl group, a benzoyl group, a pyridinecarbonyl group, a methoxycarbonyl group, a methoxycarbonylethyl group, an alkylthio group having 1 to 6 carbon atoms, a dimethylamino group, a nitro group, an acetamido group, a sulfamoyl group, a methanesulfonyl group, a benzenesulfonyl group, a pyrrolidinesulfonyl group, a morpholinesulfonyl group, a methylureido group, a butylureido group, a methoxyethylureido group, a trimethylureido group, a morpholinecarbonylamino group, and a pyridinylethoxycarbonylamino group.
The heterocyclic group refers to a saturated or unsaturated monocyclic or polycyclic heterocyclic group having 1 to 6 hetero-atoms such as an oxygen atom, a sulfur atom, and a nitrogen atom. Its examples are an imidazolyl group, a pyrazolyl group, a thiazolyl group, an oxazolyl group, an isoxazolyl group, a furyl group, a thienyl group, a pyrrolyl group, a pyridyl group, a pyrimidinyl group, a pyrazinyl group, an indolyl group, a quinolyl group, an isoquinolyl group, a benzofuranyl group, a benzothienyl group, a dihydrobenzofuranyl group, a cumarinyl group, a 2,3-dihydrobenzo[1,4]dioxinyl group, a 3,4-dihydro-2H-benzo[b][1,4]dioxepinyl group, a benzo[1,3]dioxolyl group, a 2-oxo-2,3-dihydrobenzoxazolyl group, a benzo[1,2,5]thiadiazolyl group, a 4-methyl-3,4-dihydro-2H-benzo[1,4]oxazinyl group, and a phthalimido group.
The substituted heterocyclic group refers to the above-mentioned heterocyclic group substituted by 1 to 5 substituents selected from a halogen atom, an alkyl group having 1 to 6 carbon atoms, a methoxycarbonyl group, a benzenesulfonyl group, and an oxazolyl group.
The pharmaceutically acceptable salt refers to a salt with an alkali metal, an alkaline earth metal, ammonium, an alkylammonium or the like, or a salt with a mineral acid or an organic acid. Its examples are a sodium salt, a potassium salt, a calcium salt, an ammonium salt, an aluminum salt, a triethylammonium salt, an acetate, a propionate, a butyrate, a formate, a trifluoroacetate, a maleate, a tartrate, a citrate, a stearate, a succinate, an ethylsuccinate, a lactobionate, a gluconate, a glucoheptonate, a benzoate, a methanesulfonate, an ethanesulfonate, a 2-hydroxyethanesulfonate, a benzenesulfonate, a p-toluenesulfonate, a lauryl sulfate, a malate, an aspartate, a glutamate, an adipate, a salt with cysteine, a salt with N-acetylcysteine, a hydrochloride, a hydrobromide, a phosphate, a sulfate, a hydroiodide, a nicotinate, an oxalate, a picrate, a thiocyanate, an undecanoate, a salt with an acrylic polymer, and a salt with a carboxyvinyl polymer.
The compound of the present invention may be present as a stereoisomer such as an optical isomer, a diastereomer, or a geometric isomer. All of these stereoisomers and their mixtures are included in the compounds of the present invention.
The compound of the present invention can be synthesized, for example, by the methods shown below.
(1) A compound represented by the following formula (a)
where R3 and R4 are as defined above, is reacted with a compound represented by the formula R′OH (where R′ represents an alkyl group having 1 to 6 carbon atoms) in the presence of a reagent such as trimethylsilyl chloride. Then, the product is reacted with a compound represented by the following formula (b)
where R6 is as defined above,
in a solvent, or without a solvent, in the presence of a base to obtain a compound represented by the following formula (c)
where R3, R4, R6 and R′ are as defined above.
(2) The compound represented by the formula (c) is reacted with hydrazine in a solvent or under solventless conditions to obtain a compound represented by the following formula (d)
where R3, R4 and R6 are as defined above.
(3) The resulting compound (d) is reacted with a compound represented by the following formula (e)
R2—N═C═S (e)
where R2 is as defined above,
in a solvent or under solventless conditions to obtain a compound represented by the following formula (f)
where R2, R3, R4 and R6 are as defined above.
(4) The resulting compound (f) is cyclized by a base in a solvent or under solventless conditions to obtain a compound represented by the following formula (g)
where R2, R3, R4 and R6 are as defined above.
(5) The resulting compound (g) is reacted with a compound represented by the following formula (h)
R18-L (h)
where R18, R2, R3, R4 and R6 are as defined above, can be synthesized.
(6) Also, the compound of the present invention represented by the above formula (i) is reacted with a compound represented by the following formula (j)
R51-L (j)
where R18, R2, R3, R4, R51 and R6 are as defined above.
(7) Further, the compound represented by the formula (i) or (k) above is reacted with an oxidizing agent in a solvent, whereby still another compound of the present invention represented by the following formula (l)
where R18, R2, R3, R4, R5 and R6 are as defined above, can be synthesized.
(8) Then, the compound represented by the above formula (l) is reacted with a compound represented by the formula
R18-A1-H (m)
where A1, R18, R2, R3, R4, R5 and R6 are as defined above,
can be synthesized.
Alternatively, the compound of the present invention can be synthesized by the method shown below.
(2-1) A compound represented by the following formula (o)
where R3, R4 and R″ are as defined above.
(2-2) The resulting compound of the formula (p) is reacted with a compound represented by the following formula (e)
R2—N═C═S (e)
where R2 is as defined above,
in a solvent or under solventless conditions to obtain a compound represented by the following formula (q)
where R2, R3, R4 and R″ are as defined above.
(2-3) The resulting compound of the formula (q) is cyclized by a base in a solvent or under solventless conditions to obtain a compound represented by the formula (r)
where R2, R3, R4 and R″ are as defined above.
(2-4) The resulting compound of the formula (r) is reacted with a compound represented by the following formula (h)
R18-L (h)
where R18 and L are as defined above,
in a solvent or under solventless conditions in the presence of a base to obtain a compound represented by the following formula (s)
where R18, R2, R3, R4 and R″ are as defined above.
(2-5) The resulting compound of the formula (s) is reacted with an oxidizing agent in a solvent to obtain a compound represented by the following formula (t)
where R18, R2, R3, R4 and R″ are as defined above.
(2-6) The resulting compound of the formula (t) is subjected to deprotection of the amino group under ordinary conditions, such as reaction with an acid, in a solvent to obtain a compound represented by the following formula (u)
where R18, R2, R3 and R4 are as defined above, or a salt of the compound.
(2-7) The resulting compound of the formula (u) is reacted with a compound represented by the following formula (m)
R18-A1-H (m)
where A1 and R18 are as defined above,
in a solvent or under solventless conditions in the presence of a base to carry out the formation of a salt, as appropriate, thereby obtaining a compound represented by the following formula (v)
where A1, R18, X, R2, R3 and R4 are as defined above, or a pharmaceutically acceptable salt thereof.
(2-8) The resulting compound of the formula (v) is reacted with a compound represented by the following formula (b)
where R6 is as defined above,
in a solvent or under solventless conditions in the presence of a base, whereby the compound of the present invention represented by the following formula (w)
where A1, R18, R2, R3, R4 and R6 are as defined above, can be obtained.
(2-9) The resulting compound of the present invention, represented by the formula (w), is reacted with a compound represented by the following formula (j)
R51-L (j)
where R51 and L are as defined above,
in a solvent or under solventless conditions in the presence of a base, whereby another compound of the present invention represented by the following formula (x)
where A1, R18, R2, R3, R4, R51 and R6 are as defined above, can be obtained.
Examples of the base used in the above reactions are alkali metal salts such as sodium carbonate, potassium carbonate, cesium carbonate, sodium bicarbonate, potassium bicarbonate, sodium hydroxide, dimsyl sodium, sodium hydride, sodium amide, and tert-butyl potassium, amines such as triethylamine, diisopropylamine, pyrrolidine and piperidine, sodium acetate, and potassium acetate.
Examples of the acid are inorganic acids (for example, hydrochloric acid, hydrobromic acid, hydroiodic acid, sulfuric acid and nitric acid), and organic acids (for example, trifluoroacetic acid, p-toluenesulfonic acid, and methanesulfonic acid).
As the oxidizing agent, there can be used, for example, organic peracids such as m-perchlorobenzoic acid, magnesium monoperphthalate hexahydrate, peracetic acid, and performic acid, inorganic and organic peroxides such as hydrogen peroxide, urea hydrogen peroxide adduct/phthalic anhydride, tert-butyl hydroperoxide, and cumene hydroperoxide, sodium periodate, Oxone (registered trademark), N-bromosuccinimide, N-chlorosuccinimide, Chloramine-T, tert-butyl hypochlorite, iodobenzene diacetate, and bromo-1,4-diazabicyclo[2,2,2]octane addition complex.
As the reaction solvent, there can be used, for example, water, alcohols such as methanol, ethanol, isopropyl alcohol, and tert-butyl alcohol, ethers such as dioxane and tetrahydrofuran, and solvents inert to reactions, such as dimethylformamide, N,N′-dimethylacetamide, N,N′-dimethylpropyleneurea (DMPU), hexamethylphosphoramide (HMPA), dimethyl sulfoxide, pyridine, methylene chloride, chloroform, acetone, acetic acid, and benzene.
The reaction can be performed at atmospheric pressure, under pressurized conditions, under microwave irradiation, etc. at an appropriate temperature selected within the range of −78° C. to the boiling point of the solvent for the reaction.
When the compound of the present invention is to be used as a pharmaceutical product, vehicles, bulking agents, pH regulators, solubilizers, etc. in common use are added to the compound of the present invention, the resulting blends are formed into tablets, granules, pills, capsules, powders, liquids and solutions, suspensions, injections, etc. by pharmaceutical manufacturing techniques in common use, and the resulting preparations can be administered orally or as injections or as agents for topical application.
The compound of the present invention can be administered to an adult patient in a daily dose of 1 to 1,000 mg given singly or as several divided portions. This dose can be increased or decreased, as appropriate, depending on the type of the disease, the age, body weight and symptoms of the patient, and so on.
The compound of the present invention has been found to be a potent Edg-1(S1P1) ligand, as demonstrated in the Test Examples to be described later.
The present invention will be described in further detail by the following Examples and Test Examples:
(1) Trimethylchlorosilane (12.4 ml) was added, at room temperature, to a methanol (37 ml) suspension of DL-norvaline (2.157 g), and the mixture was stirred for 2 days at room temperature, and then heated under reflux for 3 hours.
The reaction mixture was cooled to room temperature, and then the solvent was distilled off under reduced pressure. The resulting light yellow solid was dissolved in chloroform (37 ml), and triethylamine (10.3 ml) and 4-chlorobenzenesulfonyl chloride (3.886 g) were added at 0° C., followed by stirring the mixture for 2 hours at room temperature. The reaction mixture was added to an aqueous solution (120 ml) of hydrochloric acid (2 mols/liter), and the mixture was extracted with ethyl acetate (200 ml), followed by washing the extract with a saturated aqueous solution of sodium chloride (100 ml×2). The organic layer was dried over anhydrous magnesium sulfate, and then the solvent was distilled off under reduced pressure to obtain 2-{[(4-chlorophenyl)sulfonyl]amino}pentanoic acid methyl ester (4.592 g) as a light yellow oily substance.
1H NMR (300 MHz, CDCl3) δ ppm: 0.89 (t, J=7.3 Hz, 3H), 1.20-1.80 (m, 4H), 3.52 (s, 3H), 3.87-3.98 (m, 1H), 5.11 (d, J=9.5 Hz, 1H), 7.47 (d, J=8.9 Hz, 2H), 7.77 (d, J=8.9 Hz, 2H)
(2) To a methanol (50 ml) solution of the 2-{[(4-chlorophenyl)sulfonyl]amino}pentanoic acid methyl ester (4.590 g) obtained in Example 1-(1), hydrazine monohydrate (21.8 ml) was added at room temperature, and the mixture was stirred for 14 hours at room temperature. From the reaction mixture, the solvent was distilled off under reduced pressure, and water (150 ml) was added to the residue. The mixture was extracted with ethyl acetate (200 ml), and washed with a saturated aqueous solution of sodium chloride (100 ml×2).
The organic layer was dried over anhydrous magnesium sulfate, and then the solvent was distilled off under reduced pressure to obtain 4-chloro-N-[1-(hydrazinocarbonyl)butyl]benzenesulfonamide (4.368 g).
Melting point: 120.0-120.5° C.
1H NMR (300 MHz, DMSO-d6) δ ppm: 0.74 (t, J=7.3 Hz, 3H), 0.98 1.52 (m, 4H), 3.63 (t, J=7.2 Hz, 1H), 3.94 4.10 (m, 2H), 7.63 (d, J=8.9 Hz, 2H), 7.75 (d, J=8.9 Hz, 2H), 8.08 (s, 1H), 9.10 (s, 1H)
(3) Methyl isothiocyanate (683 mg) was added, at room temperature, to an ethanol (85 ml) solution of the 4-chloro-N-[1-(hydrazinocarbonyl)butyl]benzenesulfonamide (2.596 g) obtained in Example 1-(2). The mixture was stirred for 30 minutes, and then stirred for 2 hours under reflux conditions. The solvent was distilled off from the reaction mixture to obtain a solid. The solid was washed with chloroform (100 ml), and then dried to obtain 2-(2-{[(4-chlorophenyl)sulfonyl]amino}pentanoyl)-N-methylhydrazine carbothioamide (2.868 g).
Melting point: 191.0-195.0° C.
1H NMR (300 MHz, DMSO-d6) δ ppm: 0.71 (t, J=7.3 Hz, 3H), 1.00 1.64 (m, 4H), 2.88 (d, J=4.2 Hz, 3H), 3.56 3.75 (m, 1H), 7.26 7.46 (m, 1H), 7.66 (d, J=8.7 Hz, 2H), 7.83 (d, J=8.7 Hz, 2H), 9.29 (s, 1H), 10.00 (s, 1H)
(4) An aqueous solution (8.5 ml) of sodium hydroxide (1 mol/liter) was added, at room temperature, to a mixed solution in methanol (9.5 ml) and dioxane (19 ml) of the 2-(2-{[(4-chlorophenyl)sulfonyl]amino}pentanoyl)-N-methylhydrazine carbothioamide (2.157 g) obtained in Example 1-(3). The mixture was stirred for 30 minutes, and then stirred for 30 minutes at 85° C. The solvent was distilled off from the reaction mixture, and an aqueous solution (20 ml) of hydrochloric acid (0.5 mol/liter) was added to the residue. The mixture was extracted with ethyl acetate (100 ml), and washed with a saturated aqueous solution (100 ml) of sodium chloride.
The organic layer was dried over anhydrous magnesium sulfate, and then the solvent was distilled off under reduced pressure to obtain Compound 128 (2.177 g).
1H NMR (300 MHz, CDCl3) δ ppm: 0.89 (t, J=7.3 Hz, 3H), 1.20 1.95 (m, 4H), 3.58 (s, 3H), 4.44 4.57 (m, 1H), 6.59 6.76 (m, 1H), 7.45 (d, J=8.5 Hz, 2H), 7.69 (d, J=8.5 Hz, 2H), 11.41 (s, 1H)
Diisopropylamine (0.407 ml) and allyl bromide (0.218 ml) were added, at room temperature, to a tetrahydrofuran (9.7 ml) solution of the 4-chloro-N-[1-(5-mercapto-4-methyl-4H-1,2,4-triazol-3-yl)butyl]benzenesulfonamide (Compound 128) (698 mg) obtained in Example 1-(4), and the mixture was stirred overnight at room temperature. The solvent was distilled off from the reaction mixture under reduced pressure. The resulting residue was dissolved in ethyl acetate (100 ml), and the solution was washed sequentially with an aqueous solution (50 ml) of hydrochloric acid (1 mol/liter) and a saturated aqueous solution (100 ml) of sodium chloride. The organic layer was dried over anhydrous magnesium sulfate, and then the solvent was distilled off under reduced pressure. The residue was recrystallized from a solvent mixture of hexane (20 ml) and ethyl acetate (15 ml) for purification, thereby obtaining Compound 125 (590 mg).
Melting point: 161.5-162.0° C.
1H NMR (300 MHz, CDCl3) δ ppm: 0.86 (t, J=7.3 Hz, 3H), 1.15 1.35 (m, 2H), 1.65 1.96 (m, 2H), 3.39 (s, 3H), 3.73 3.80 (m, 2H), 4.37 4.49 (m, 1H), 5.07 5.27 (m, 2H), 5.85 6.01 (m, 2H), 7.40 (d, J=8.9 Hz, 2H), 7.70 (d, J=8.9 Hz, 2H)
Mesyl chloride (25.8 mg) and triethylamine (0.055 ml) were added to a chloroform (0.9 ml) solution of benzyl alcohol (16.2 mg), and the mixture was stirred for 3 hours at room temperature. The reaction mixture was eluted by NH-type silica gel (Chromatorex, Fuji Silysia Chemical Ltd.) column chromatography using tetrahydrofuran as a solvent to obtain methanesulfonic acid benzyl ester. To a tetrahydrofuran (0.9 ml) solution of the methanesulfonic acid benzyl ester, 4-chloro-N-[(1R)-1-(4-ethyl-5-mercapto-4H-1,2,4-triazol-3-yl)ethyl]benzenesulfonamide (Compound 90) (17.3 mg) obtained by performing the same procedure as in Example 1 using the corresponding raw materials, and potassium t-butoxide (8.4 mg) were added, followed by stirring the mixture for 20 hours at 40° C. PSA (polymer supported amine) (0.15 ml) was added to the reaction mixture, and the resulting mixture was stirred for 4 hours at room temperature. The reaction mixture was eluted with ethyl acetate and tetrahydrofuran, and the solvents were distilled off. The resulting residue was eluted by NH-type silica gel column chromatography using tetrahydrofuran as a solvent, and the eluate was purified by silica gel column chromatography to obtain Compound 5 (2.3 mg).
1H NMR (200 MHz, DMSO-d6) δ ppm: 1.10 (t, J=7.1 Hz, 3H), 1.25 (d, J=6.8 Hz, 3H), 3.62 3.96 (m, 2H), 4.36 (s, 2H), 4.55 4.78 (m, 1H), 7.15 7.47 (m, 5H), 7.64 (d, J=8.6 Hz, 2H), 7.77 (d, J=8.6 Hz, 2H), 8.52 (d, J=8.6 Hz, 1H)
To a chloroform (18 ml) solution of 4-chloro-N-{(1R)-1-[4-ethyl-5-(methylthio)-4H-1,2,4-triazol-3-yl]ethyl}benzenesulfonamide (Compound 1) (329 mg) obtained by performing the same procedure as in Examples 1 and 2 using the corresponding raw materials, m-perchlorobenzoic acid (157 mg) was added under ice-cooling conditions. The mixture was stirred for 30 minutes at 0° C., and then stirred overnight at room temperature. The reaction solution was washed with a saturated aqueous solution (20 ml) of sodium bicarbonate. The organic layer was dried over anhydrous magnesium sulfate, and then the solvent was distilled off under reduced pressure. The resulting residue was eluted by silica gel flash column chromatography using a solvent mixture of acetone and ethyl acetate. The eluate was developed twice by preparative TLC (Merck, analytical TLC plate, 20×20 cm, Silicagel 60F254×4 plates) using ethyl acetate as a solvent. Then, the developed substances were eluted using a 5% methanol/chloroform mixed solution to obtain Compound 180 (14 mg) of low polarity and Compound 181 (21 mg) of high polarity (Compound 180 and Compound 181: diastereomers).
Compound 180 of low polarity: 1H NMR (300 MHz, CDCl3) δ ppm: 1.47 (t, J=7.2 Hz, 3H), 1.54 (d, J=6.8 Hz, 3H), 3.26 (s, 3H), 4.18 4.46 (m, 2H), 4.63 4.78 (m, 1H), 5.95 (d, J=9.6 Hz, 1H), 7.45 (d, J=8.5 Hz, 2H), 7.70 (d, J=8.5 Hz, 2H)
Compound 181 of high polarity: 1H NMR (300 MHz, CDCl3) δ ppm: 1.47 (t, J=7.3 Hz, 3H), 1.55 (d, J=6.8 Hz, 3H), 3.24 (s, 3H), 4.18 4.49 (m, 2H), 4.63 4.80 (m, 1H), 6.00 (d, J=9.3 Hz, 1H), 7.46 (d, J=8.8 Hz, 2H), 7.72 (d, J=8.8 Hz, 2H)
Compound 5
To a chloroform (30 ml) solution of 4-chloro-N-{(1R)-1-[4-ethyl-5-(methylthio)-4H-1,2,4-triazol-3-yl]ethyl}benzenesulfonamide (Compound 1) (520 mg) obtained by performing the same procedure as in Examples 1 and 2 using the corresponding raw materials, m-perchlorobenzoic acid (746 mg) was added under ice-cooling conditions, followed by stirring the mixture for 1 hour at 0° C. A saturated aqueous solution (100 ml) of sodium bicarbonate was added to the reaction mixture, and the resulting mixture was extracted with chloroform (50 ml). The organic layer was dried over anhydrous magnesium sulfate, and then the solvent was distilled off under reduced pressure. The resulting residue was purified by silica gel flash column chromatography using a solvent mixture of ethyl acetate and chloroform to obtain Compound 182 (481 mg).
1H NMR (300 MHz, CDCl3) δ ppm: 1.46 (t, J=7.3 Hz, 3H), 1.54 1.60 (m, 3H), 3.49 (s, 3H), 4.25 4.40 (m, 2H), 4.65 4.78 (m, 1H), 5.44 (d, J=9.8 Hz, 1H), 7.45 (d, J=8.9 Hz, 2H), 7.68 (d, J=8.9 Hz, 2H)
To a dimethylformamide (2 ml) solution of N-{(1R)-1-(5-allylthio)-4-ethyl-4H-1,2,4-triazol-3-yl}ethyl)-4-chlorobenzenesulfonamide (Compound 3) (200 mg) obtained by performing the same procedure as in Examples 1 and 2 using the corresponding raw materials, potassium carbonate (120 mg) and methyl iodide (0.040 ml) were added at room temperature, followed by stirring the mixture for 3 hours at room temperature. Ethyl acetate was added to the reaction mixture, and the resulting mixture was washed sequentially with a 1 mol/liter aqueous solution of hydrochloric acid and a saturated aqueous solution of sodium chloride. The organic layer was dried over anhydrous magnesium sulfate, and then the solvent was distilled off under reduced pressure. The resulting residue was eluted by silica gel flash column chromatography using a solvent mixture of ethyl acetate and hexane. Then, the eluate was recrystallized using a solvent mixture of ethyl acetate and hexane for purification, whereby Compound 98 (112 mg) was obtained.
Melting point: 142.0-143.0° C.
1H NMR (200 MHz, DMSO-d6) δ ppm: 1.13 (d, J=6.8 Hz, 3H), 1.27 (t, J=7.1 Hz, 3H), 2.56 (s, 3H), 3.82 (d, J=7.1 Hz, 2H), 3.90 4.25 (m, 2H), 5.04 5.27 (m, 2H), 5.43 (q, J=6.8 Hz, 1H), 5.82 6.06 (m, 1H), 7.74 (d, J=8.8 Hz, 2H), 7.90 (d, J=8.8 Hz, 2H)
Sodium hydride (12 mg) was added, while cooled with ice, to a dimethylformamide (1.2 ml) solution of the 4-chloro-N-{(1R)-1-[4-ethyl-5-(methylsulfonyl)-4H-1,2,4-triazol-3-yl]ethyl}benzenesulfonamide (Compound 182) (47 mg) obtained in Example 5 and n-propanol (0.027 ml). The mixture was stirred for 30 minutes at room temperature, and then stirred for 2 hours at 100° C. After the reaction mixture was cooled to room temperature, it was added to a saturated aqueous solution (5 ml) of ammonium chloride. The mixture was extracted with ethyl acetate (20 ml×2) and washed with a saturated aqueous solution (20 ml) of sodium chloride. The organic layer was dried over anhydrous magnesium sulfate, and then the solvent was distilled off under reduced pressure. The resulting residue was purified by silica gel flash column chromatography using a solvent mixture of methanol and chloroform to obtain Compound 183 (35 mg).
1H NMR (300 MHz, CDCl3) δ ppm: 1.02 (t, J=7.5 Hz, 3H), 1.24 (t, J=7.2 Hz, 3H), 1.47 (d, J=6.8 Hz, 3H), 1.75 1.91 (m, 2H), 3.68 3.80 (m, 2H), 4.37 (t, J=6.5 Hz, 2H), 4.45 4.58 (m, 1H), 5.60 (bs, 1H), 7.45 (d, J=8.6 Hz, 2H), 7.77 (d, J=8.6 Hz, 2H)
(1) Hydrazine monohydrate (30 ml) was added to a methanol (180 ml) solution of N-(t-butoxycarbonyl)-D-alanine methyl ester (41.8 g), and the mixture was stirred for 12 hours at room temperature. The reaction mixture was concentrated, and the resulting crude crystals were washed with a solvent mixture of hexane and ethyl acetate (1:1, 300 ml). Then, the washed crystals were dried to obtain (R)-(1-hydrazinocarbonyl-2-ethyl)carbamic acid t-butyl ester as a colorless powder (32.6 g).
1H NMR (300 MHz, DMDO-d6) δ ppm: 1.14 (d, J=7.2 Hz, 3H), 1.37 (s, 9H), 3.30-4.09 (m, 3H), 6.70-6.90 (m, 1H), 8.96 (br s, 1H)
(2) Ethyl isothiocyanate (14.6 ml) was added to an EtOH (152 ml) solution of the (R)-(1-hydrazinocarbonyl-2-ethyl)carbamic acid t-butyl ester (30.8 g) obtained in Example 8-(1), and the mixture was heated for 2 hours under reflux. After the reaction mixture was cooled to room temperature, crystals precipitated were filtered. The filtrate was concentrated, and the resulting residue was purified by silica gel chromatography using a solvent mixture of ethyl acetate and chloroform, whereby (R)-2-(N-(t-butoxycarbonyl)amino)propionyl)-N-ethylhydrazinecarbothioamide was obtained as a colorless amorphous substance (43.2 g).
1H NMR (300 MHz, DMSO-d6) δ ppm: 0.98-1.28 (m, 6H), 1.40 (s, 9H), 3.25-3.65 (m, 2H), 3.77-3.95 (m, 1H), 7.20-7.39 (m, 1H), 7.45-7.60 (m, 1H), 9.25 (s, 1H), 10.00 (s, 1H)
(3) An aqueous solution (218 ml) of sodium hydroxide (1 mol/liter) was added to a mixed solution, in methanol (120 ml) and dioxane (240 ml), of the (R)-2-(N-(t-butoxycarbonyl)amino)propionyl)-N-ethylhydrazinecarbothioamide (42.1 g) obtained in Example 8-(2), followed by heating the mixture for 3 hours under reflux. The reaction mixture was concentrated, and an aqueous solution (100 ml) of hydrochloric acid (2 mols/liter) was added. The mixture was extracted with an ethyl acetate-CHCl3-MeOH mixed solution (10:10:1, 500 ml), and the organic layer was dried over anhydrous magnesium sulfate. Then, the solvent was distilled off under reduced pressure, and the resulting residue was washed with a solvent mixture of hexane and ethyl acetate (1:1, 300 ml), followed by drying, to obtain [(R)-1-(4-ethyl-5-mercapto-4H-[1,2,4]triazol-3-yl)ethyl]-carbamic acid t-butyl ester as a white solid (29.22 g).
1H NMR (300 MHz, DMSO-d6) δ ppm: 1.21 (t, J=7.1 Hz, 3H), 1.30-1.50 (m, 3H), 1.39 (s, 9H), 3.82-4.05 (m, 2H), 4.72-4.88 (m, 1H), 7.58 (d, J=8.5 Hz, 1H), 13.60 (br s, 1H)
(4) Diisopropylamine (17.4 ml) and iodomethane (7.7 ml) were added to a tetrahydrofuran (200 ml) solution of the [(R)-1-(4-ethyl-5-mercapto-4H-[1,2,4]triazol-3-yl)ethyl]-carbamic acid t-butyl ester (28.12 g) obtained in Example 8-(3). The mixture was stirred for 1 hour at room temperature, and then precipitated crystals were filtered. The filtrate was concentrated, and the resulting crude crystals were washed with a hexane-ethyl acetate solvent mixture (3:1, 200 ml), followed by drying, to obtain [(R)-1-(4-ethyl-5-methylsulfanyl-4H-[1,2,4]triazol-3-yl)ethyl]-carbamic acid t-butyl ester as a white powder (29.5 g).
1H NMR (300 MHz, DMSO-d6) δ ppm: 1.21 (t, J=7.0 Hz, 3H), 1.38 (s, 9H), 1.45 (t, J=7.0 Hz, 3H), 2.62 (s, 3H), 3.80-4.00 (m, 2H), 4.85-4.92 (m, 1H), 7.52 (d, J=8.5 Hz, 1H)
(5) To a chloroform (293 ml) solution of the [(R)-1-(4-ethyl-5-methylsulfanyl-4H-[1,2,4]triazol-3-yl)ethyl]-carbamic acid t-butyl ester (21.0 g) obtained in Example 8-(4), m-perchlorobenzoic acid (43.0 g) was added in four portions while being cooled with ice. The mixture was stirred for 3 hours at room temperature, and then stirred for 1 hour at 40° C. To the reaction mixture, Na2S2O3 (12.9 g) and an aqueous solution (300 ml) of sodium hydroxide (1 mol/liter) were added. The organic layer was separated, and washed with a saturated aqueous solution of sodium chloride. The organic layer was dried over anhydrous magnesium sulfate, and then the solvent was distilled off under reduced pressure. The resulting residue was purified by silica gel flash column chromatography using a solvent mixture of hexane and ethyl acetate. Then, the purified substance was recrystallized using hexane and chloroform to obtain [(R)-1-(4-ethyl-5-methanesulfonyl-4H-[1,2,4]triazol-3-yl)ethyl]-carbamic acid t-butyl ester as a white powder (17.2 g).
1H NMR (300 MHz, CDCl3) δ ppm: 1.44 (s, 9H), 1.49 (t, J=7.1 Hz, 3H), 1.67 (t, J=6.8 Hz, 3H), 3.53 (s, 3H), 4.25-4.59 (m, 2H), 4.92-5.20 (m, 2H)
(6) Trifluoroacetic acid (121 ml) was added to the [(R)-1-(4-ethyl-5-methanesulfonyl-4H-[1,2,4]triazol-3-yl)ethyl]-carbamic acid t-butyl ester (100.0 g) obtained in Example 8-(5), and the mixture was stirred for 2 hours at room temperature. The reaction mixture was concentrated under reduced pressure to obtain (R)-1-(4-ethyl-5-methanesulfonyl-4H-[1,2,4]triazol-3-yl)ethylamine trifluoroacetate as a white powder (103.8 g).
1H NMR (300 MHz, DMSO-d6) δ ppm: 1.37 (t, J=7.2 Hz, 3H), 1.59 (t, J=6.8 Hz, 3H), 3.65 (s, 3H), 4.21-4.50 (m, 2H), 4.72-4.90 (m, 1H), 8.69 (br s, 3H)
(7) NaOMe (18 ml, 2.0N, MeOH solution) was added to the (R)-1-(4-ethyl-5-methanesulfonyl-4H-[1,2,4]triazol-3-yl)ethylamine trifluoroacetate (3.0 g) obtained in Example 8-(6), and the mixture was heated for 1 hour under reflux. The reaction mixture was cooled to room temperature, and Et2O (100 ml) was added. After the mixture was cooled to 0° C., precipitated crystals were filtered. The filtrate was concentrated, and the resulting crude product was purified by NH silica gel chromatography using a solvent mixture of Et2O and MeOH to obtain the captioned compound (Compound 519) as a colorless oily substance (1.55 g).
1H NMR (300 MHz, CDCl3) δ ppm: 1.32 (t, J=7.2 Hz, 3H), 1.54 (t, J=6.7 Hz, 3H), 3.78-3.95 (m, 2H), 4.02-4.20 (m, 1H), 4.13 (s, 3H)
A tetrahydrofuran (0.9 ml) solution of benzenesulfonyl chloride (31 mg) was added, at room temperature, to the compound (20 mg) obtained in Example 8-(7). Then, triethylamine (0.040 ml) was added, and the mixture was stirred for 3 hours at room temperature. The reaction mixture was eluted by NH-type silica gel column chromatography using tetrahydrofuran as a solvent, and then the eluate was concentrated to obtain the captioned compound (36.5 mg).
1H NMR (200 MHz, CDCl3) δ ppm: 1.24 (t, J=7.1 Hz, 3H), 1.41 (t, J=7.0 Hz, 3H), 3.79 (q, J=7.1 Hz, 3H), 4.09 (s, 3H), 4.40-4.67 (m, 1H), 6.20-6.60 (m, 1H), 7.40-7.62 (m, 3H), 7.80-7.98 (m, 2H)
Using the same methods as the methods used in Examples 1 to 9, salts were formed, as appropriate, to obtain the compounds shown in the tables offered below. As Compounds 89, 104, 136 and 137, those purchased from Bionet were used.
The compounds obtained in the above-described Examples 1 to 9 are also shown in Table 1 along with the other compounds.
Using a human Edg-1(S1P1) gene transferred HEK-293 cell strain (showing a binding of Kd=6.4±2.1 nM, Bmax=160±94 fmol/105 cells to [3H]-S1P), which had been obtained by a method complying with the method described in the literature (Science. 1998, 279:1552), the Edg-1(S1P1) binding inhibiting effect of the compound of the present invention was investigated in accordance with the method described in the literature. Cells (1×105 cells/well) obtained as described above were seeded in poly-L-lysine-coated 96-well plates (Corning Incorporated), and then incubated in a 5% carbon dioxide gas incubator for 12 hours at 37° C. with the use of an MEM medium (Invitrogen Corporation) containing 100 U/mL of penicillin, 100 μg/mL of streptomycin, a 1% MEM nonessential amino acid solution, and 10% FCS. The cultured cells were washed twice with a buffer (20 mM Tris-HCl, pH 7.4, 100 mM NaCl, 15 mM NaF, 2 mM deoxypyridoxine, 4 mg/mL fatty acid-free BSA), and was then treated with 100 μL of the buffer incorporating [3H]-S1P (produced by ARC, end concentration 10 nM) and a DMSO solution of the test compound (end concentration of the compound: 10−5 M, end concentration of DMSO: 0.1%) for 1 hour at 4° C. After the cells were washed twice with the buffer, they were solubilized with 100 μL of Opti Phase Supermix (produced by Perkin-Elmer), and measured for radioactivity by means of Micro Beta (produced by Perkin-Elmer). Based on the radioactivity, the amount (A) of binding of [3H]-S1P upon the addition of the compound was calculated.
The same procedure was performed in the absence of the test compound, and the amount (B) of binding of [3H]-S1P was calculated. Moreover, the same procedure was performed in the absence of the test compound with the use of HEK-293 cells, to which the Edg-1(S1P1) gene had not been transferred, and the background amount (C) of binding of [3H]-S1P was calculated.
The Edg-1(S1P1) binding inhibition rate of the compound, calculated from the following equation, is shown in Table 1.
Inhibition rate(%)=[1−(A−C)/(B−C)]×100
Using a human Edg-1(S1P1) gene transferred HEK-293 cell strain membrane fraction, the Edg-1(S1P1) binding inhibiting effect of the compound of the present invention was investigated in accordance with the method described in the literature (Science. 2002, 296:346) (showing a binding of Kd=0.15 nM, Bmax=2.5 fmol/μg to [33P]-S1P). The membrane fraction was obtained by treating the cells with a solubilizing buffer (1 mM Tris/HCl, pH 7.2) for 10 minutes on ice, centrifuging the system (1000×g, 5 min) to remove insoluble fractions, and then centrifuging the system (40000×g, 30 min, 4° C.). The resulting membrane fraction was dissolved in a binding buffer (20 mM Tris-HCl, pH 7.4, 100 mM NaCl, 15 mM NaF, 2 mM deoxypyridoxine, 4 mg/mL fatty acid-free BSA), and then [33P]-S1P (produced by ARC, end concentration 0.1 nM) and a DMSO solution of the test compound (end concentration of the compound: 10−5 M, end concentration of DMSO: 0.1%) were added, followed by stirring the mixture and subsequently treating it for 1 hour at 30° C. Using a harvester, the membrane fraction was harvested onto unifilter-96GF/C filter (produced by Perkin-Elmer). Then, the filter with the membrane fraction was washed 4 times with the binding buffer, and the filter was dried. To the filter, 25 μL Microscint 0 (produced by Perkin-Elmer) was added, and the assay system was measured for radioactivity by means of Top Count NXT (Packard). Based on the radioactivity, the amount (A) of binding of [33P]-S1P to the membrane fraction upon addition of the compound was calculated.
The same procedure was performed in the absence of the test compound, and the amount (B) of binding of [33P]-S1P was calculated. Moreover, the same procedure was performed in the absence of the test compound with the use of HEK-293 cells, to which the Edg-1(S1P1) gene had not been transferred, and the background amount (C) of binding of [33P]-S1P was calculated.
The Edg-1(S1P1) binding inhibition rate of the compound, calculated from the following equation, is shown in
1H NMR
The compound of the present invention is an excellent Edg-1(S1P1) ligand. Hence, it is useful as an agent for the treatment or prevention of autoimmune diseases such as Crohn disease, irritable colitis, Sjögren syndrome, multiple sclerosis, and systemic lupus erythematosus, and diseases such as rheumatoid arthritis, asthma, atopic dermatitis, rejection reaction after organ transplantation, cancer, retinopathy, psoriasis, osteoarthrosis, and age-related macular degeneration.
| Number | Date | Country | Kind |
|---|---|---|---|
| 2004-28394 | Aug 2004 | JP | national |
| 2005-121769 | Apr 2005 | JP | national |
| Filing Document | Filing Date | Country | Kind | 371c Date |
|---|---|---|---|---|
| PCT/JP05/14351 | 8/4/2005 | WO | 00 | 2/1/2007 |
