Patent Application
20110159019
The present invention relates to a 2,4-diaminopyrimidine compound which is useful as an active ingredient for a pharmaceutical composition, in particular, a pharmaceutical composition for inhibiting acute rejection occurring in transplantation.
Protein kinase C (PKC) is one of the protein kinase families, of which at least ten kinds of isozymes have hitherto been identified and which have been classified into three subfamilies according to differences in the primary structures.
The activation mechanisms of these three subfamilies are greatly different among the subfamilies. A type of PKC which is activated by calcium and diacyl glycerol (DAG) is called a classical PKC (cPKC), and a type of PKC which is activated by DAG but which does not need calcium in this activation is called a novel PKC (nPKC) and a type of PKC which does not need either calcium or DAG is called atypical PKC (aPKC).
Furthermore, each subfamily consists of plural isozymes, cPKC is classified into PKCα, PKCβ and PKCγ, nPKC is classified into PKCδ, PKCε, PKCη and PKCθ, and aPKC is classified into PKCξ and PKCλ.
The expression distribution of each isozyme covers a relatively wide range, but the expression of a PKCθ which is one nPKC is restricted to the T lymphocytes and the skeletal muscles. In addition, the phenotype of knockout mice of PKCθ exhibits inhibition of T cell signaling or induction of T cell anergy, and further, from the viewpoint that abnormalities of the skeletal muscles are not observed, PKCθ is promising as a target of an immunosuppressor having few side-effects.
Moreover, since PKCθ is in complementary relationship in the T cell receptor signaling pathway with calcineurin, which is a target molecule of FK506 and cyclosporin A, which have been widely used in current transplantation medication, there is a possibility that combination use of a calcineurin inhibitor and a PKCθ inhibitor will express a synergic immunosuppressive effect.
Therefore, it is considered that if PKCθ is inhibited selectively, an immunosuppressive activity is expressed with a low level of side-effects, and as a transplantation medication, it is promising in regard to the inhibition of acute rejection occurring in transplantation, and also, there is a possibility that it will be able to express a synergic immunosuppressive activity when used in combination with a calcineurin inhibitor.
In Patent Citation 1, it is reported that a compound represented by the formula (A) inhibits PKCθ and is useful as an immunosuppressor. As a specific compound, a compound having a pyrimidine structure is disclosed, but there is no specific disclosure of the compound of the present invention.
(R2 in the formula represents
or the like. For the other symbols, reference can be made to the publication.)
In Patent Citation 2, it is reported that a compound represented by the formula (B) inhibits PKCθ and is useful as an immunosuppressor. As a specific compound, a compound having a pyrimidine structure is disclosed, but there is no specific disclosure of the compound of the present invention.
(R3 in the formula represents
For the other symbols, reference can be made to the publication.)
In Patent Citation 3, it is reported that a compound represented by the formula (C) inhibits PKCθ and is useful as an immunosuppressor. As a specific compound, a compound having a pyrimidine structure is disclosed, but there is no specific disclosure of the compound of the present invention.
(R1 in the formula represents
For the other symbols, reference can be made to the publication.)
In Patent Citation 4, it is reported that a compound represented by the formula (D) has an inhibition activity against a cyclin-dependent kinase (CDK), a kinase of Aurora B, or the like, and is useful for treatment and prevention of diseases characterized by excessive or abnormal cell growth. As a specific compound, a compound having a pyrimidine structure is disclosed and there is a description that the compound is useful for immunosuppression in organ transplantation, but there is no specific disclosure of the compound of the present invention.
(For the symbols in the formula, reference can be made to the publication.)
In Patent Citation 5, it is reported that a compound represented by the formula (E) inhibits a polo-like kinase (PLK) and is thus useful for prevention and/or treatment of diseases associated with tumors, neurodegenerative diseases, and activation of immune systems. As a specific compound, a compound having a pyrimidine structure is disclosed, but there is no specific disclosure of the compound of the present invention. Also, there is neither description of technologies concerning a PKCθ inhibition activity nor description that the compound is useful for inhibition of acute rejection occurring in transplantation.
(For the symbols in the formula, reference can be made to the publication.)
In Patent Citation 6, it is reported that a compound represented by the formula (F) inhibits a G protein-coupled receptor protein 88 (GPR88) and is thus useful for prevention and/or treatment of central nervous system diseases. As a specific compound, a compound having a pyrimidine structure is disclosed, but there is no specific disclosure of the compound of the present invention. Also, there is neither description of technologies concerning a PKCθ inhibition activity nor description that the compound is useful for inhibition of acute rejection occurring in transplantation.
(R1 in the formula represents hydrogen or the like and A represents a heterocyclic group which may be substituted, heterocyclic alkyl which may be substituted, C3-8 cycloalkyl which may be substituted, or the like. For the other symbols, reference can be made to the publication.)
It is an object of the present invention to provide a 2,4-diaminopyrimidine compound which is useful as an active ingredient of a pharmaceutical having a PKCθ inhibition activity, particularly, a pharmaceutical composition for inhibiting acute rejection occurring in transplantation.
The present inventors have conducted extensive studies on a compound having a PKCθ inhibition activity, and as a result, they have found that a compound having a structure such as aralkyl and the like on an amino group at the 2-position and also having a structure such as an adamantylalkyl group and the like on an amino group at the 4-position of 2,4-diaminopyrimidine, or a salt thereof has an excellent PKCθ inhibition activity, thereby completing the present invention.
Thus, the present invention relates to a compound of the formula (I) or a pharmaceutically acceptable salt thereof, and a pharmaceutical composition comprising the compound of the formula (I) or a pharmaceutically acceptable salt thereof and a pharmaceutically acceptable excipient.
(the symbols in the formula have the following meanings:
R1 represents any one group selected from the group consisting of:
R4 represents —OH, amine which may be substituted, or —CH2NH2;
n1 represents 0 or 1;
R5 represents —OH, (C1-6 alkyl which may be substituted with —OH or —NH2), or —CN;
R6 represents —H or C1-6 alkyl which may be substituted with aryl;
p represents 0 or 1;
q represents 1, 2, 3, or 4;
R13 represents —H or C1-6 alkyl;
R2 represents —CN, —CF3, —NO2, or halogen;
A represents a single bond or C1-6 alkylene;
R3 represents any one group selected from the group consisting of:
R9s are the same as or different from each other and represent halogen, C1-6 alkyl which may be substituted, —OH, —CN, cycloalkyl, -Q-(C1-6 alkyl which may be substituted), or aryl which may be substituted;
Q represents —O—, —S—, —SO—, —SO2—, or —NHSO2—;
n2 represents 0, 1, 2, or 3;
R10 represents halogen, C1-6 alkyl, —CN, —O—(C1-6 alkyl), —S—(C1-6 alkyl), —SO—(C1-6 alkyl), —SO2—(C1-6 alkyl), —S-(cycloalkyl), or —OCF3; and
R12 represents —H or halogen).
In this regard, when a symbol in a certain chemical formula in this specification is used in a different chemical formula, the same symbol has the same meaning, unless otherwise indicated.
In addition, the present invention relates to a pharmaceutical composition comprising the compound of the formula (I) or a pharmaceutically acceptable salt thereof, and a pharmaceutically acceptable excipient, for inhibiting acute rejection occurring in transplantation; i.e., an agent for inhibiting acute rejection occurring in transplantation, comprising the compound of the formula (I) or a pharmaceutically acceptable salt thereof, and a pharmaceutically acceptable excipient.
Moreover, the present invention relates to use of the compound of the formula (I) or a pharmaceutically acceptable salt thereof for the manufacture of an inhibitor of acute rejection occurring in transplantation, and a method for inhibiting acute rejection occurring in transplantation, comprising administering to a patient an effective amount of the compound of the formula (I) or a pharmaceutically acceptable salt thereof.
The compound of the formula (I) or a pharmaceutically acceptable salt thereof has a PKCθ inhibition action and can be used as an inhibitor of acute rejection occurring in transplantation.
According to the present invention, the following are provided.
[1]
A compound of the formula (I) or a pharmaceutically acceptable salt thereof:
(the symbols in the formula have the following meanings:
R1 represents any one group selected from the group consisting of:
R4 represents —OH, amine which may be substituted, or —CH2NH2;
n1 represents 0 or 1;
R5 represents —OH, (C1-6 alkyl which may be substituted with —OH or —NH2), or —CN;
R6 represents —H or C1-6 alkyl which may be substituted with aryl;
p represents 0 or 1;
q represents 1, 2, 3, or 4;
R13 represents —H or C1-6 alkyl;
R2 represents —CN, —CF3, —NO2, or halogen;
A represents a single bond or C1-6 alkylene;
R3 represents any one group selected from the group consisting of:
R9s are the same as or different from each other and represent halogen, C1-6 alkyl which may be substituted, —OH, —CN, cycloalkyl, -Q-(C1-6 alkyl which may be substituted), or aryl which may be substituted;
Q represents —O—, —S—, —SO—, —SO2—, or —NHSO2—;
n2 represents 0, 1, 2, or 3;
R10 represents halogen, C1-6 alkyl, —CN, —O—(C1-6 alkyl), —S—(C1-6 alkyl), —SO—(C1-6 alkyl), —SO2—(C1-6 alkyl), —S-(cycloalkyl), or —OCF3; and
R12 represents —H or halogen).
[2]
The compound or a pharmaceutically acceptable salt thereof described in [1],
wherein
R4 is —OH, —NR7R8, or —CH2NH2;
R7 and R8 are the same as or different from each other and represent:
(a) —H;
(b) C1-6 alkyl, in which the C1-6 alkyl may be substituted with at least one group selected from the group consisting of the following 1) to 12):
1) —OH
2) protected —OH
3) halogen
4) —COOH
5) —CONH2
6) oxo
7) aryl
8) heteroaryl
9) cycloalkyl which may be substituted with at least one group selected from the group consisting of —OH, protected —OH, (C1-6 alkyl which may be substituted with —OH), halogen, —CN, NR14R15, —CONR14R15, —SO2NR14R15, (C1-6 alkyl which may be substituted with —OH)—O—, and oxo
10) heterocycloalkyl which may be substituted with —OH or (C1-6 alkyl which may be substituted with —OH, —OCH3, —CN, or halogen)
11) (heterocycloalkyl which may be substituted with —OH or —NH2)—CO—, and
12) (heterocycloalkyl)-NH—CO—;
(c) cycloalkyl, in which the cycloalkyl may be substituted with at least one group selected from the group consisting of the following 1) to 6):
1) —OH
2) —NHR11
3) halogen
4) oxo
5) C1-6 alkyl which may be substituted with —OH, and
6) heterocycloalkyl which may be substituted with (halogen, —OH, —CH2OH, or —COCH3);
(d) heterocycloalkyl, in which the heterocycloalkyl may be substituted with at least one group selected from the group consisting of the following 1) to 11):
1) C1-6 alkyl which may be substituted with (—OH, —OCH3, —CN, halogen, or —CONH2)
2) cycloalkyl
3) aryl
4) heterocycloalkyl
5) heterocycloalkyl-CO—
6) —COCH3
7) —CONH2
8) —COCH2OH
9) —COOCH2CH3
10) —SO2CH3
11) oxo, and
12) halogen;
(e) aryl;
(f) nicotinoyl; and
(g) —SO2CH3; or
(h) R7 and R8, together with a nitrogen atom to which they bind, are a nitrogen-containing a heterocycloalkyl which may be substituted with at least one group selected from the group consisting of (—OH, —NH2, —COOH, —COCH3, —CONH2 and —CH2OH);
R11 is —H, C1-6 alkyl which may be substituted with (halogen or —OH), cycloalkyl which may be substituted with halogen, heterocycloalkyl which may be substituted with —COCH3, or —COCH3; and
R14 and R15 are the same as or different from each other and are —H, C1-6 alkyl, or heterocycloalkyl.
[3]
The compound or a pharmaceutically acceptable salt thereof described in [2],
wherein
R1 is
and
R3 is
[4]
The compound or a pharmaceutically acceptable salt thereof described in [3],
wherein
R4 is —NR7R8;
R7 and R8 are the same as or different from each other and are
(b) C1-6 alkyl, in which the C1-6 alkyl may be substituted with at least one group selected from the group consisting of the following 1) to 12):
1) —OH
2) —OH protected with methyl group, or when having two OH groups adjacent to each other, —OH protected with a dimethylmethylene group or a benzylidene group
3) —F
4) —COOH
5) —CONH2
6) oxo
7) phenyl
8) pyridyl
9) cyclohexyl which may be substituted with at least one group selected from the group consisting of —OH and (C1-6 alkyl which may be substituted with —OH)
10) (piperidinyl or pyrrolidinyl) which may be substituted with —OH or (C1-6 alkyl which may be substituted with —OH, —OCH3, —CN, or —F)
11) (piperazinyl)-CO— or (piperidinyl which may be substituted with —OH or —NH2)—CO—, and
12) (piperidinyl)-NH—CO—; or
(c) cycloalkyl, in which the cycloalkyl may be substituted with at least one group selected from the group consisting of the following 1) to 6):
1) —OH
2) —NHR11
3) —F
4) oxo
5) C1-6 alkyl which may be substituted with —OH, and
6) (azetidinyl, pyrrolidinyl, piperidinyl, piperazinyl or morpholinyl) which may be substituted with (halogen, —OH, —CH2OH, or —COCH3); is —H;
n1 is 1;
R2 is —CN, —CF3, —NO2, or —F;
A is C1-6 alkylene;
R9 is
(i) —F, —Cl, or —Br
(j) C1-6 alkyl which may be substituted with —OH or halogen,
(k) —OH,
(l) —CN,
(m) cyclopropyl,
(n) -Q-(C1-6 alkyl which may be substituted with halogen, —OH, —OCH3, —CN, or —CONH2), or
(o) phenyl which may be substituted with —CH2NH2; and n2 is 1.
[5]
The compound or a pharmaceutically acceptable salt thereof described in [4],
wherein
R7 and R8 are the same as or different from each other and are
(b) C1-6 alkyl, in which the C1-6 alkyl may be substituted with the following groups:
9) cyclohexyl substituted with at least one group selected from the group consisting of —OH, —CH3, and —CH2OH, and
10) piperidinyl which may be substituted with —OH or (C1-6 alkyl which may be substituted with —OH, —OCH3, —CN, or —F); or
(c) cycloalkyl, in which the cycloalkyl may be substituted with at least one group selected from the group consisting of the following 1), 2), and 5):
1) —OH
2) —NHR11, and
5) C1-6 alkyl which may be substituted with —OH;
R11 is —H;
R2 is —CN;
A is methylene;
R9 is
(i) —F, —Cl, or —Br
(j) C1-6 alkyl which may be substituted with —OH or —F,
(k) —OH,
(l) —CN,
(m) cyclopropyl,
(n) -Q-(C1-6 alkyl which may be substituted with halogen, —OH, —OCH3, —CN, or —CONH2), or
(o) phenyl which may be substituted with —CH2NH2; and
R10 is —Cl, —CH3, —OCH3, —OCH2CH3, —OCH(CH3)2, —SCH3, —SCH2CH3, —SCH(CH3)2, —SOCH3, —SO2CH3, —S-(cyclopentane), or —OCF3.
[6]
A pharmaceutical composition comprising the compound or a pharmaceutically acceptable salt thereof described in [1], and a pharmaceutically acceptable excipient.
[7]
A PKCθ inhibitor comprising the compound or a pharmaceutically acceptable salt thereof described in [1].
[8]
A pharmaceutical composition for inhibiting acute rejection occurring in transplantation, comprising the compound or a pharmaceutically acceptable salt thereof described in [1].
[9]
Use of the compound or a pharmaceutically acceptable salt thereof described in [1] for the manufacture of an inhibitor of acute rejection occurring in transplantation.
[10]
A method for inhibiting acute rejection occurring in transplantation, comprising administering to a patient an effective amount of the compound or a pharmaceutically acceptable salt thereof described in [1].
Hereinbelow, the present invention will be described in detail.
In the present specification, the “C1-6 alkyl” is linear or branched alkyl having 1 to 6 carbon atoms, and examples thereof include a methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, sec-butyl, tert-butyl, n-pentyl, n-hexyl, and the like.
In the present specification, the “C1-6 alkylene” is linear or branched C1-6 alkylene, and examples thereof include methylene, ethylene, trimethylene, tetramethylene, pentamethylene, hexamethylene, propylene, methylmethylene, ethylethylene, 1,2-dimethylethylene, 1,1,2,2-tetramethylethylene, and the like. In another embodiment, it is C1 alkylene, in a further embodiment, C1-2 alkylene, and in a still further embodiment, methylene or ethylene.
In the present specification, the “halogen” means F, Cl, Br, or I.
In the present specification, the “cycloalkyl” is a C3-10 saturated hydrocarbon ring group, which may have a bridge. Examples thereof include cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, cycloheptyl, cyclooctyl, adamantyl, and the like. In another embodiment, it is C3-8 cycloalkyl, in a further embodiment, C3-6 cycloalkyl, and in a still further embodiment, cyclopropyl, cyclobutyl, cyclopentyl, or cyclohexyl.
In the present specification, the “aryl” is a C6-14 monocyclic to tricyclic aromatic hydrocarbon ring group, and examples thereof include phenyl and naphthyl, and in another embodiment, phenyl.
In the present specification, the “heterocyclic ring” is a ring group selected from i) a monocyclic 3- to 8-membered heterocyclic ring, and preferably, 5- to 7-membered heterocyclic ring, containing 1 to 4 heteroatoms selected from oxygen, sulfur, and nitrogen, and ii) a bicyclic to tricyclic heterocyclic ring group containing 1 to 5 heteroatoms selected from oxygen, sulfur, and nitrogen, formed by condensation with one or two rings in which the monocyclic heterocyclic ring group is selected from the group consisting of a monocyclic heterocyclic ring group, a benzene ring, C5-8 cycloalkane, and C5-8 cycloalkene. The ring atom, sulfur or nitrogen, may be oxidized to form an oxide or a dioxide.
Examples of the “heterocyclic ring” include the following embodiments.
(1) Monocyclic Saturated Heterocyclic Ring
(a) those containing 1 to 4 nitrogen atoms, for example, azepanyl, diazepanyl, aziridinyl, azetidinyl, pyrrolidinyl, imidazolylidinyl, piperidyl, pyrazolidinyl, piperazinyl, azocanyl, and the like;
(b) those containing 1 to 3 nitrogen atoms and 1 to 2 sulfur atoms and/or 1 to 2 oxygen atoms, for example, thiomorpholinyl, thiazolidinyl, isothiazolidinyl, oxazolidinyl, morpholinyl, and the like;
(c) those containing 1 to 2 sulfur atoms, for example, tetrahydro-2H-thiopyranyl and the like;
(d) those containing 1 to 2 sulfur atoms and 1 to 2 oxygen atoms, for example, oxathiolanyl and the like; and
(e) those containing 1 to 2 oxygen atoms, for example, oxiranyl, oxetanyl, dioxolanyl, tetrahydrofuranyl, tetrahydro-2H-pyranyl, 1,4-dioxanyl, and the like;
(2) Monocyclic Unsaturated Heterocyclic Group
(a) those containing 1 to 4 nitrogen atoms, for example, pyrrolyl, imidazolyl, pyrazolyl, pyridyl, dihydropyridyl, tetrahydropyridinyl, pyrimidinyl, pyrazinyl, pyridazinyl, triazolyl, tetrazolyl, triazinyl, dihydrotriazinyl, azepinyl, and the like;
(b) those containing 1 to 3 nitrogen atoms and 1 to 2 sulfur atoms and/or 1 to 2 oxygen atoms, for example, thiazolyl, isothiazolyl, thiadiazolyl, dihydrothiazinyl, oxazolyl, isoxazolyl, oxadiazolyl, oxazinyl, and the like;
(c) those containing 1 to 2 sulfur atoms, for example, thienyl, thiepinyl, dihydrodithiopyranyl, dihydrodithionyl, and the like;
(d) those containing 1 to 2 sulfur atoms and 1 to 2 oxygen atoms, for example, dihydroxathiopyranyl and the like; and
(e) those containing 1 to 2 oxygen atoms, for example, furyl, pyranyl, oxepinyl, dioxolyl, and the like;
(3) Condensed Polycyclic Saturated Heterocyclic Group
(a) those containing 1 to 5 nitrogen atoms, for example, quinuclidinyl, 7-azabicyclo[2.2.1]heptyl, 3-azabicyclo[3.2.2]nonanyl, and the like;
(b) those containing 1 to 4 nitrogen atoms and 1 to 3 sulfur atoms and/or 1 to 3 oxygen atoms, for example, trithiadiazaindenyl, dioxoloimidazolidinyl, and the like; and
(c) those containing 1 to 3 sulfur atoms and/or 1 to 3 oxygen atoms, for example, 2,6-dioxabicyclo[3.2.2]octo-7-yl, and the like;
(4) Condensed Polycyclic Unsaturated Heterocyclic Ring Group
(a) those containing 1 to 5 nitrogen atoms, for example, indolyl, isoindolyl, indolinyl, indolidinyl, benzoimidazolyl, dihydrobenzoimidazolyl, tetrahyzorobenzimidazolyl, quinolyl, tetrahydroquinolyl, isoquinolyl, tetrahydroisoquinolyl, indazolyl, imidazopyridyl, benzotriazolyl, tetrazolopyridazinyl, carbazolyl, acridinyl, quinoxalinyl, dihydroquinoxalinyl, tetrahydroqunioxalinyl, phthalazinyl, dihydroindazolyl, benzopyrimidinyl, naphthyridinyl, quinazolinyl, cinnolinyl, and the like;
(b) those containing 1 to 4 nitrogen atoms and 1 to 3 sulfur atoms and/or 1 to 3 oxygen atoms, for example, benzothiazolyl, dihydrobenzothiazolyl, benzothiadiazolyl, imidazothiazolyl, imidazothiadiazolyl, benzoxazolyl, dihydrobenzoxazolyl, dihydrobenzoxazinyl, benzoxadiazolyl, benzoisothiazolyl, benzoisoxazolyl, and the like;
(c) those containing 1 to 3 sulfur atoms, for example, benzothienyl, benzodithiopyranyl, dibenzo[b,d]thienyl, and the like;
(d) 1 to 3 sulfur atoms and 1 to 3 oxygen atoms, for example, benzoxathiopyranyl, phenoxadinyl, and the like; and
(e) those containing 1 to 3 oxygen atoms, for example, benzodioxolyl, benzofuranyl, dihydrobenzofuranyl, isobenzofuranyl, chromanyl, chromenyl, dibenzo[b,d]furanyl, methylenedioxyphenyl, ethylenedioxyphenyl, and the like; etc.
In the present specification, the “heterocycloalkyl” is the monocyclic saturated heterocyclic ring group described in (1) and the condensed polycyclic saturated heterocyclic ring group described in (3) among the above-described “heterocyclic rings”, in which a ring atom, sulfur or nitrogen, may be oxidized to form an oxide or a dioxide. In another embodiment, it is the monocyclic saturated heterocyclic ring group described in (1), in which a ring atom, sulfur or nitrogen, may be oxidized to form an oxide or a dioxide, and in a further embodiment, it is azetidinyl, pyrrolidinyl, piperidinyl, piperazinyl, morpholinyl, tetrahydro-2H-thiopyranyl, tetrahydrothiopyranyl dioxide, or tetrahydro-2H-pyranyl.
In the present specification, the “nitrogen-containing heterocycloalkyl” is the monocyclic saturated heterocyclic ring containing at least one nitrogen atom described in (1) (a) and (b), and the condensed polycyclic saturated heterocyclic ring group containing at least one nitrogen atom described in (3) (a) and (b), among the above-described “heterocyclic rings”. In another embodiment, the nitrogen-containing heterocycloalkyl is the monocyclic saturated heterocyclic ring containing at least one nitrogen atom described in (1) (a) and (b), and in a further embodiment, azetidinyl, pyrrolidinyl, piperidyl, piperazinyl, or morpholinyl.
In the present specification, the “heteroaryl” is the heterocyclic ring having an aromaticity among (2) the monocyclic unsaturated heterocyclic ring group and (4) the aromatic heterocyclic ring group among the condensed polycyclic unsaturated heterocyclic ring groups of the above-described “heterocyclic ring”. In another embodiment, it is the heterocyclic ring having an aromaticity among (2) the aromatic heterocyclic ring group, (monocyclic heteroaryl), and in a further embodiment, pyridyl.
In the present specification, the expression “which may be substituted” means which is not substituted or which has 1 to 5 substituents, and in another embodiment, which is not substituted or which has 1 to 3 substituents. Further, the expression “(which is) substituted” means which has 1 to 5 substituents, and in another embodiment, which has 1 to 3 substituents. Furthermore, if it has a plurality of substituents, the substituents may be the same as or different from each other.
The “protected —OH” means that the OH group is protected with a protecting group usually used for the protection of a hydroxyl group. In another embodiment, it means being protected with an acyl group, an ether group, a silyl ether group, or an acetal group, and in a further embodiment, it means protection with a methyl group or in the case that two OH groups are adjacent to each other, protection with a dimethylmethylene group or a benzylidene group.
Embodiments regarding the compound (I) of the present invention are shown below.
(1) The compound, wherein R1 is
(2) The compound as described in (1), wherein R4 is —OH, —NR7R8, or —CH2NH2, and in another embodiment, —NR7R8.
(3) The compound as described in (2), wherein R7 and R8 are the same as or different from each other and are
(a) —H;
(b) C1-6 alkyl, in which the C1-6 alkyl may be substituted with at least one group selected from the group consisting of the following 1) to 12):
1) —OH
2) protected —OH, and in another embodiment, —OH protected with a methyl group, or in the case of having two OH groups which are adjacent to each other, —OH protected with a dimethylmethylene group or a benzylidene group
3) halogen, and in another embodiment, —F
4) —COOH
5) —CONH2
6) oxo
7) aryl, and in another embodiment, phenyl
8) heteroaryl, and in another embodiment, pyridyl
9) cycloalkyl which may be substituted with at least one group selected from the group consisting of —OH, protected —OH, (C1-6 alkyl which may be substituted with —OH), halogen, —CN, —NR14R15, —CONR14R15, —SO2NR14R15, (C1-6 alkyl which may be substituted with —OH)—O—, and oxo, in another embodiment, cyclohexyl which may be substituted with at least one group selected from the group consisting of —OH and (C1-6 alkyl which may be substituted with —OH), and in a further embodiment, cyclohexyl substituted with at least one group selected from the group consisting of —OH, —CH3, and —CH2OH
10) heterocycloalkyl which may be substituted with —OH or (C1-6 alkyl which may be substituted with —OH, —OCH3, —CN, or halogen), and in another embodiment, (piperidinyl or pyrrolidinyl) which may be substituted with —OH or (C1-6 alkyl which may be substituted with —OH, —OCH3, —CN, or F)
11) (heterocycloalkyl which may be substituted with —OH or —NH2)—CO—, and in another embodiment, (piperazinyl)-CO— or (piperidinyl which may be substituted with —OH or —NH2)—CO—, and
12) (heterocycloalkyl)-NH—CO—, and in another embodiment, (piperidine)-NH—CO—;
(c) cycloalkyl, and in another embodiment, cyclobutyl or cyclohexyl, in which the cycloalkyl may be substituted with at least one group selected from the group consisting of the following 1) to 6):
1) —OH
2) —NHR11
3) halogen, and in another embodiment, —F
4) oxo
5) C1-6 alkyl which may be substituted with —OH, and in another embodiment, —CH3 or —CH2OH, and
6) heterocycloalkyl which may be substituted with (halogen, —OH, —CH2OH, or —COCH3), and in another embodiment, (azetidinyl, pyrrolidinyl, piperidinyl, piperazinyl, or morpholinyl) which may be substituted with (—F, —OH, —CH2OH or —COCH3);
(d) heterocycloalkyl, and in another embodiment, azetidinyl, piperidinyl, tetrahydro-2H-pyranyl, or tetrahydro-2H-thiopyranyl, in which the heterocycloalkyl may be substituted with at least one group selected from the group consisting of the following 1) to 11):
1) C1-6 alkyl which may be substituted with (—OH, —OCH3, —CN, halogen, or —CONH2), and in another embodiment, C1-6 alkyl which may be substituted with (—OH, —OCH3, —CN, —F, or —CONH2)
2) cycloalkyl, and in another embodiment, cyclopropyl
3) aryl, and in another embodiment, phenyl
4) heterocycloalkyl, and in another embodiment, tetrahydro-2H-pyranyl
5) heterocycloalkyl-CO—, and in another embodiment, morpholinyl-CO—
6) —COCH3
7) —CONH2
8) —COCH2OH
9) —COOCH2CH3
10) —SO2CH3
11) oxo, and
12) halogen;
(e) aryl, and in another embodiment, phenyl;
(f) nicotinoyl; and
(g) —SO2CH3; or
(h) R7 and R8, together with a nitrogen atom to which they bind, are a nitrogen-containing heterocycloalkyl which may be substituted with at least one group selected from the group consisting of (—OH, —NH2, —COOH, —COCH3, —CONH2 and —CH2OH), and in another embodiment, (azetidinyl, pyrrolidinyl, piperidinyl, or piperazinyl) which may be substituted with at least one group selected from the group consisting of (—OH, —NH2, —COOH, —COCH3, —CONH2, and —CH2OH).
(4) The compound as described in (3), wherein R11 is —H, C1-6 alkyl which may be substituted with (halogen or —OH), cycloalkyl which may be substituted with halogen, heterocycloalkyl which may be substituted with —COCH3, or —COCH3, in another embodiment, —H, C1-6 alkyl which may be substituted with (—F or —OH), cycloalkyl which may be substituted with —F, heterocycloalkyl which may be substituted with —COCH3, or —COCH3, and in a further embodiment, C1-6 alkyl which may be substituted with (—F or —OH), cyclohexyl which may be substituted with —F, tetrahydro-2H-pyranyl, piperidinyl substituted with —COCH3, or —COCH3.
(5) The compound as described in (3), wherein R14 and R15 are the same as or different from each other and are —H, C1-6 alkyl, or heterocycloalkyl, and in another embodiment, —H, methyl, or tetrahydro-2H-pyranyl.
(6) The compound as described in (1), wherein n1 is 1.
(7) The compound, wherein R5 is —OH, —CH2OH, —CH2NH2, or —CN.
(8) The compound, wherein R6 is —H or C1-6 alkyl which may be substituted with aryl, and in another embodiment, —H or C1-6 alkyl which may be substituted with phenyl.
(9) The compound, wherein R2 is —CN.
(10) The compound, wherein A is C1-6 alkylene, in another embodiment, methylene or ethylene, and in a further embodiment, methylene.
(11) The compound, wherein R3 is
(12) The compound as described in (11), wherein R9s are the same as or different from each other and are
(i) halogen, and in another embodiment, —F, —Cl, or —Br;
(j) C1-6 alkyl which may be substituted, in another embodiment, C1-6 alkyl which may be substituted with —OH or halogen, and in a further embodiment, C1-6 alkyl which may be substituted with —OH or —F;
(k) —OH;
(l) —CN;
(m) cycloalkyl, and in another embodiment, cyclopropyl;
(n) -Q-(C1-6 alkyl which may be substituted), and in another embodiment, -Q-(C1-6 alkyl which may be substituted with halogen, —OH, —OCH3, —CN, or —CONH2); or
(o) aryl which may be substituted, in another embodiment, aryl which may be substituted with —CH2NH2, and in a further embodiment, phenyl which may be substituted with —CH2NH2, and
(13) The compound as described in (11), wherein n2 is 1.
(14) The compound as described in (11), wherein R10 is —Cl, —CH3, —OCH3, —OCH2CH3, —OCH(CH3)2, —SCH3, —SCH2CH3, —SCH(CH3)2, —SOCH3, —SO2CH3, —S-(cyclopentane), or —OCF3, and in another embodiment, —Cl, —CH3, —OCH3, or —SCH3.
(15) The compound, wherein R12 is —H or —Cl.
(16) The compound, which is a combination of two or more of the groups as described (1) to (15) above.
The compound of the formula (I) may exist in the form of tautomeric properties or geometrical isomers in some cases, depending on the kind of substituents. In the present specification, the compound shall be described in only one form of isomer, yet the present invention includes other isomers, isolated forms of the isomers, or a mixture thereof.
In addition, the compound of the formula (I) may have asymmetric carbon atoms or axial chirality in some cases, and correspondingly, it may exist in the form of optical isomers. The present invention includes both an isolated form of the optical isomers of the compound of the formula (I) or a mixture thereof
In addition, the pharmaceutically acceptable prodrugs of the compound represented by the formula (I) are also included in the present invention. The pharmaceutically acceptable prodrug refers to a compound which is converted into the compound of the present invention by solvolysis or under a physiological condition. Examples of the group for forming a prodrug include those as described in Prog. Med., 5, 2157-2161 (1985) or “Iyakuhin no Kaihatsu (Development of Medicines)” (Hirokawa Shoten, 1990), Vol. 7, Bunshi Sekkei (Molecular Design), 163-198.
Furthermore, the compound of the formula (I) refers to a pharmaceutically acceptable salt of the compound of the formula (I), and it may form a salt with an acid or a base, depending on the kind of the substituents. Specifically, examples thereof include acid addition salts with inorganic acids such as hydrochloric acid, hydrobromic acid, hydroiodic acid, sulfuric acid, nitric acid, phosphoric acid, and the like, and with organic acids such as formic acid, acetic acid, propionic acid, oxalic acid, malonic acid, succinic acid, fumaric acid, maleic acid, lactic acid, malic acid, mandelic acid, tartaric acid, dibenzoyl tartaric acid, ditoluoyl tartaric acid, citric acid, methanesulfonic acid, ethanesulfonic acid, benzenesulfonic acid, p-toluenesulfonic acid, aspartic acid, glutamic acid, and the like, and salts with inorganic bases such as sodium, potassium, magnesium, calcium, aluminum, and the like, and organic bases such as methylamine, ethylamine, ethanolamine, lysine, ornithine, and the like, salts with various amino acids or amino acid derivatives such as acetylleucine and the like, ammonium salts, and others.
In addition, the present invention also includes various hydrates or solvates, and polymorphic crystal substances of the compound of the formula (I) and a pharmaceutically acceptable salt thereof. Also, the present invention includes compounds labeled with various radioactive or non-radioactive isotopes.
(Preparation Methods)
The compound of the formula (I) and a pharmaceutically acceptable salt thereof can be prepared by applying various known synthesis methods, using the characteristics based on their basic skeletons or the kind of substituents. At this time, depending on the type of the functional groups, it is in some cases effective, from the viewpoint of the preparation techniques, to substitute the functional group with an appropriate protecting group (a group which is capable of being easily converted into the functional group), during the steps from starting materials to intermediates. Examples of such a protective group include those described in “Green's Protective Groups in Organic Synthesis (4th Edition, 2006)”, edited by Wuts (P. G. M. Wuts) and Greene (T. W. Green), which may be appropriately selected and used depending on reaction conditions. In these methods, a desired compound can be obtained by introducing the protecting group to carry out the reaction, and then, if desired, removing the protecting group.
In addition, the prodrug of the compound of formula (I) can be prepared by introducing a specific group during the steps from starting materials to intermediates, in the same manner as for the aforementioned protecting groups, or by carrying out the reaction using the obtained compound of formula (I). The reaction can be carried out by applying a method known to a person skilled in the art, such as general esterification, amidation, dehydration, and the like.
Hereinbelow, the representative preparation methods for the compound of formula (I) will be described. Each of the production processes may also be carried out with reference to the References appended in the present description. Further, the preparation methods of the present invention are not limited to the examples as shown below.
Production Process 1
(wherein Lv1 and Lv2 represent a leaving group. The same shall apply hereinafter.)
The present production process is a method in which a compound (1) and an amine compound (2) are subjected to a nucleophilic substitution reaction to prepare a compound (3), and the obtained compound (3) and an amine compound (4) are subjected to a nucleophilic substitution reaction to prepare the compound (I) of the present invention. Here, examples of the leaving group include halogen, a methanesulfonyloxy group, a methylsulfinyl group, a methylsulfonyl group, a p-toluenesulfonyloxy group, and the like.
In this reaction, the compound (1) and the compound (2), or the compound (3) and the compound (4) are used in equivalent amounts or with either thereof in an excess amount, and the mixture is stirred under any temperature condition from cooling to heating with reflux in a solvent which is inert to the reaction or without a solvent, preferably at 0° C. to 80° C., usually for 0.1 hour to 5 days. The solvent used herein is not particularly limited, but examples thereof include aromatic hydrocarbons such as benzene, toluene, xylene, and the like, ethers such as diethyl ether, tetrahydrofuran, dioxane, dimethoxyethane, and the like, halogenated hydrocarbons such as dichloromethane, 1,2-dichloroethane, chloroform, and the like, N,N-dimethylformamide, N-methylpyrrolidone, 1,3-dimethyl-2-imidazolidinone, dimethylsulfoxide, ethyl acetate, acetonitrile, and a mixture thereof. It may be advantageous in some cases for the smooth progress of the reaction to carry out the reaction in the presence of organic bases such as triethylamine, N,N-diisopropylethylamine, N-methylmorpholine, and the like, or inorganic bases such as potassium carbonate, sodium carbonate, potassium hydroxide, and the like. In this regard, the compound (2) may be reacted after reacting the compound (1) and the compound (4) first.
Production Process 2: Other Production Processes
Moreover, several compounds represented by the formula (I) can be prepared from the compound of the formula (I) of the present invention obtained above, by any combination of the processes that can be generally employed by a person skilled in the art, such as well-known amidation, alkylation, reductive amination, reduction of a carbonyl group to a hydroxyl group, and the like. For example, they can be prepared, for example, by the reactions as described below, the methods as described in Examples to be described later, the methods known to a skilled person in the art, or a modified method thereof.
2-1: Amidation
An amide compound can be obtained by subjecting a carboxylic acid compound and an amine compound to amidation.
In this reaction, a carboxylic acid compound and an amine compound are used in equivalent amounts, or with either thereof in an excess amount, and the mixture thereof is stirred at any temperature from under cooling to heating, preferably at a temperature from −20° C. to 60° C., usually for 0.1 hour to 5 days, in a solvent which is inert to the reaction, in the presence of a condensing agent. Examples of the solvent as used herein are not particularly limited, and include aromatic hydrocarbons such as benzene, toluene, xylene, and the like, halogenated hydrocarbons such as dichloromethane, 1,2-dichloroethane, chloroform, and the like, ethers such as diethyl ether, tetrahydrofuran, dioxane, dimethoxyethane, and the like, N,N-dimethylformamide, dimethylsulfoxide, ethyl acetate, acetonitrile, water, and a mixture thereof. Examples of the condensing agent include 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide, dicyclohexylcarbodiimide, 1,1′-carbonyldiimidazole, diphenylphosphoric azide, and phosphorus oxychloride, but are not limited thereto. It may be preferable for the reaction in some cases to use an additive (for example, 1-hydroxybenzotriazole). It may be advantageous in some cases for the smooth progress of the reaction to carry out the reaction in the presence of an organic base such as triethylamine, N,N-diisopropylethylamine, N-methylmorpholine, and the like, or an inorganic base such as potassium carbonate, sodium carbonate, potassium hydroxide, and the like.
Further, a method in which the carboxylic acid is converted into a reactive derivative thereof, and then the reactive derivative is reacted with the amine compound may also be used. Examples of the reactive derivative of the carboxylic acid include acid halides obtained by the reaction of a halogenating agent such as phosphorus oxychloride, thionyl chloride, and the like, mixed acid anhydrides obtained by the reaction of isobutyl chloroformate or the like, active esters obtained by the condensation with 1-hydroxybenzotriazole or the like, etc. The reaction of the reactive derivative and the amine compound can be carried out at any temperature from under cooling to heating, preferably at −20° C. to 60° C., in a solvent which is inert to the reaction, such as halogenated hydrocarbons, aromatic hydrocarbons, ethers, and the like.
2-2: Alkylation
An alkyl amine compound can be prepared by alkylating the amine compound with a compound having a leaving group.
The alkylation can be carried out by the same method as in Production Process 1.
2-3: Reductive Amination
An amine compound can be alkylated by reducing an imine compound which is prepared from a carbonyl compound and a primary or secondary amine compound.
In this reaction, the carbonyl compound and the primary or secondary amine compound are used in equivalent amounts, or with either thereof in an excess amount, and the mixture thereof is stirred at any temperature from under cooling to heating, preferably at a temperature from −45° C. to heating under reflux, and preferably at 0° C. to room temperature, usually for 0.1 hour to 5 days, in a solvent which is inert to the reaction, in the presence of a reducing agent. Examples of the solvent as used herein are not particularly limited, and include alcohols such as methanol, ethanol, and the like, halogenated hydrocarbons such as dichloromethane, 1,2-dichloroethane, chloroform, and the like, ethers such as diethyl ether, tetrahydrofuran, dioxane, dimethoxyethane, and the like, and a mixture thereof. Examples of the reducing agent include sodium cyanoborohydride, sodium triacetoxyborohydride, sodium borohydride, and the like. The reaction may be preferably carried out in the presence of a dehydrating agent such as molecular sieves and the like, or an acid such as acetic acid, hydrochloric acid, titanium (IV) isopropoxide complexes, and the like in some cases. According to the reaction, there may be some cases where an imine is produced by the condensation of the carbonyl compound with the primary or secondary amine compound and it can be isolated as a stable intermediate. In this case, the imine intermediate can be isolated, and then subjected to a reduction reaction, thereby obtaining a desired product. Further, the reaction can be carried out in a solvent such as methanol, ethanol, ethyl acetate, and the like, in the presence or absence of an acid such as acetic acid, hydrochloric acid, and the like, using a reduction catalyst (for example, palladium on carbon, Raney nickel, and the like), instead of treatment with the reducing agent. In this case, it is preferable to carry out the reaction under a hydrogen atmosphere at normal pressure to 50 atmospheres from under cooling to under heating.
2-4: Reduction of Carbonyl Group to Hydroxyl Group
An alcohol compound can be obtained by subjecting a carbonyl compound to reduction.
In this reaction, the carbonyl compound is treated with an equivalent amount or an excess amount of a reducing agent at any temperature from under cooling to heating, preferably at a temperature from −20° C. to 80° C., usually for 0.1 hour to 3 days, in a solvent which is inert to the reaction. Examples of the solvent as used herein are not particularly limited, and include ethers such as diethyl ether, tetrahydrofuran, dioxane, dimethoxyethane, and the like, alcohols such as methanol, ethanol, 2-propanol, and the like, aromatic hydrocarbons such as benzene, toluene, xylene, and the like, N,N-dimethylformamide, dimethylsulfoxide, ethyl acetate, and a mixture thereof. As the reducing agent, hydride reducing agents such as sodium borohydride, diisobutylaluminum hydride, and the like, metal reducing agents such as sodium, zinc, iron, and the like, or others described in the following documents are suitably used.
(Production Process for Starting Compound)
The starting materials used in the preparation of the compound of the present invention, that is, the amine compound (2) and the amine compound (4) can be prepared, for example, from available well-known compounds, by employing the methods described in Preparation Examples as described later, well-known methods described in “Production Process 2: Other Production Processes”, or methods apparent to a skilled person in the art, or modified methods thereof, or the like.
The compound of the formula (I) is isolated and purified as a free compound, pharmaceutically acceptable salts thereof, hydrates, solvates, or polymorphic crystal substances thereof. The pharmaceutically acceptable salt of the compound of the formula (I) can also be prepared in accordance with a conventional method for a salt formation reaction.
Isolation and purification are carried out by employing general chemical operations such as extraction, fractional crystallization, various types of fraction chromatography, and the like.
Various isomers can be separated by selecting an appropriate starting compound or by making use of the difference in the physicochemical properties between isomers. For example, the optical isomer can be derived into a stereochemically pure isomer by means of general optical resolution methods (for example, fractional crystallization for inducing diastereomer salts with optically active bases or acids, chromatography using a chiral column and the like, and others). In addition, the isomers can also be prepared from an appropriate optically active starting compound.
The pharmacological activity of the compound of the formula (I) was confirmed by the following test.
Test Method 1: Measurement of Human PKCθ Enzyme Inhibition Activity
The test was carried out using a HTRF® KinEASE™ S1 kit (CIS bio). To a 384-well plate (CORNING) were put 4 μL of a liquid agent and 3 μL of a mixed liquid of STK Substrate 1-biotin (final 250 nM), and Full-length human PKCθ (Carna Biosciences, final 31 ng/mL), followed by leaving it to stand at room temperature for 30 minutes. Then, 3 μL of an ATP liquid (final 30 μM) was dispensed therein to carry out an enzyme reaction at room temperature for 1 hour. Thereafter, the reaction was stopped by addition of 10 μL of a solution of Sa-XL665 (final 31.25 nM) and an antibody STK-Antibody-Cryptate (finally 800-fold diluted), and the mixture was left to stand at room temperature for 1 hour Fluorescence intensities at 620 nm (Cryptate) and 665 nm (XL665) were measured in Discovery (PACKARD), and with reference to a Vehicle at 0% inhibition and a Blank of 100% inhibition, the inhibition rates and IC50 values were calculated.
The test results are shown in Table 1. Ex represents Compound No. of Examples as described later.
Test Method 2: Measurement of Human IL-2 Production Inhibition Activity
i) Preparation of Plasmid
The DNA fragments (445 bp) in the Human IL-2 promoter region corresponding to the DNA sequence as described in the database were cloned and inserted into pGL3 basic which is a Vector for Reporter Gene Assay to acquire pGL3-IL2-pro-43.
ii) Maintenance/Passage of Jurkat Cells
Jurkat, Clone E6-1 (ATCC No. TIB-152), which is a human T cell-based culture cell was cultured under the conditions of 37° C., 5% CO2, and saturated humidity, using 10% FBS RPMI 1640 (Sigma) as a medium, and at a time point of a confluency of about 90%, passage was carried out.
iii) Transfection and Seeding
A cell suspension of a concentration of 2.5×107 cells/mL was prepared using 10% FBS RPMI 1640 (Sigma) by counting the cells using a cell counting plate, and 10 μg of pGL3-IL2-pro-43 was mixed therewith. Then, 400 μL of the Jurkat cells prepared at 2.5×107 cells/mL were added to each of the prepared plasmid mixture and mixed, followed by adding it entirely to Gene Pulsor® Cuvette (BIO-RAD). By Gene Pulsor®II (BIO-RAD), a plasmid was introduced at 300 V and 975 μF, and the whole amount of the Jurkat cells having the plasmid introduction completed were gently suspended in 2.5 mL of 10% FBS RPMI 1640. Then, the cells were seeded to a 96-well plate (Corning Coster) at 50 μL/well, and cultured for about 10 hours under the condition of 37° C., 5% CO2, and saturated humidity.
iv) Measurement of Human IL-2 Production Inhibition Activity
A drug solution was added respectively at 25 μL/well, and additionally, a mixed liquid obtained by 250-fold dilution of an anti-CD3 antibody, an anti-CD28 antibody (Pharmingen) (all 1000-fold liquid of the final concentration of 1 μg/mL) with 10% FBS RPMI1640 was added respectively thereto at 25 μL/well. The resultant was cultured for about 14 hours under the condition of 37° C., 5% CO2, and saturated humidity. The assay was performed in duplicate.
A substrate solution supplied by a Bright-Gol™ Luciferase Assay System (Promega) was added respectively at 100 μL/well and mixed gently. A Multilabel Counter (ARVO SX, WALLAC) was set at a reaction temperature: 25° C., Shaking Duration: 1 sec, and Measurement time: 1 sec, the measurement well of each of the 96-well plates was set up, and a Firefly luciferase activity was measured.
Test Method 3: Measurement of Cytochrome P450 (CYP3A4) Enzyme Inhibition Activity
i) Inhibition Test I (Calculation of Remaining Rate I)
Using a 96-well plate, 2 μM of a substrate (midazolam), 5 μM of a test compound, and human liver microsome (0.1 mg protein/mL) were incubated at 37° C. for 20 minutes in a total amount of 150 μL of a 100 mM phosphate buffer (pH 7.4) containing 0.1 mM EDTA and 1 mM NADPH. Then, the reaction was stopped by adding 130 μl of an aqueous solution containing 80% acetonitrile. Thereafter, the samples were analyzed by LC/MS/MS, and the remaining rates I were calculated using the following equation 1.
Remaining Rate I(%)=Ai,I/Ao,I×100 (Equation 1)
Ai, I=Amount of produced metabolite after reaction in the presence of the test compound in the inhibition test I
Ao, I=Amount of produced metabolite after reaction in the absence of the test compound in the inhibition test I
ii) Inhibition Test II (Calculation of Remaining Rate II)
Using a 96-well plate, 5 μM of a test compound and human liver microsome (0.1 mg protein/mL) were incubated at 37° C. for 30 minutes in a total amount of 145 μL of a 100 mM phosphate buffer (pH=7.4) containing 0.1 mM EDTA and 1 mM NADPH. Then, 2 μM of midazolam as the substrate was added thereto at a total amount of 150 μL of and incubated at 37° C. for 20 minutes. After the incubation, the reaction was stopped by adding 130 μL of an aqueous solution containing 80% acetonitrile. Thereafter, the samples were analyzed by LC/MS/MS, and the remaining rate II was calculated using the following equation 2.
Remaining Rate II(%)=Ai,II/Ao,II/(Ai,I/Ao,I)×100 (Equation 2)
Ai, II=Amount of produced metabolite after reaction in the presence of the test compound in the inhibition test II
Ao, I=Amount of produced metabolite after reaction in the absence of the test compound in the inhibition test II
The test results are shown in Table 2. Ex represents No. of the Example Compounds as described below.
As a result of each of the above tests, the compound of the formula (I) has a PKCθ inhibition action and reduction in CYP inhibition, from which it is apparent that the compound is useful for an inhibitor of acute rejection occurring in transplantation, or the like.
A pharmaceutical composition containing one or two or more kinds of the compound of formula (I) or a pharmaceutically acceptable salt thereof as an active ingredient can be prepared in accordance with a generally used method, using a pharmaceutical excipient, a pharmaceutical carrier, or the like, that is generally used in the art.
Administration may be carried out through any mode of oral administration via tablets, pills, capsules, granules, powders, liquid preparations, or the like, or parenteral administration via injections such as intraarticular, intravenous, intramuscular, and the like, suppositories, eye drops, eye ointments, transdermal liquid preparations, ointments, transdermal patches, transmucosal liquid preparations, transmucosal patches, inhalations, and the like.
Regarding solid composition for oral administration, tablets, powders, granules, or the like are used. In such a solid composition, one or more active ingredients are mixed with at least one inactive excipient, for example, lactose, mannitol, glucose, hydroxypropylcellulose, microcrystalline cellulose, starch, polyvinyl pyrrolidone, aluminum magnesium metasilicate, or the like. According to a conventional method, the composition may contain inactive additives, for example, a lubricant such as magnesium stearate and the like, a disintegrator such as sodium carboxymethylstarch and the like, a stabilizer, and a solubilizing agent. As occasion demands, tablets or pills may be coated with a sugar coating, or a gastric or enteric coating agent.
The liquid composition for oral administration includes pharmaceutically acceptable emulsions, solutions, suspensions, syrups, elixirs, or the like, and contains a generally used inert diluent, such as purified water or ethanol. In addition to the inert diluent, the liquid composition may contain adjuvants such as a solubilizing agent, a moisturizing agent, and a suspending agent, a sweetener, a flavor, an aromatic, and an antiseptic.
Injections for parenteral administration include sterile, aqueous or non-aqueous solutions, suspensions, or emulsions. As the aqueous solvent, for example, distilled water for injection or physiological saline is included. Examples of the non-aqueous solvent include propylene glycol, polyethylene glycol, vegetable oils such as olive oil and the like, alcohols such as ethanol and the like, polysorbate 80 (Pharmacopeia), etc. Such a composition may further contain a tonicity agent, an antiseptic, a moistening agent, an emulsifying agent, a dispersing agent, a stabilizer, or a solubilizing agent. These are sterilized, for example, by filtration through a bacteria-retaining filter, blending with bactericides, or irradiation. In addition, these can also be used by producing a sterile solid composition, and dissolving or suspending it in sterile water or a sterile solvent for injection prior to its use.
Examples of the agent for external use include ointments, plasters, creams, jellies, patches, sprays, lotions, eye drops, eye ointments, and the like. The agents contain generally used ointment bases, lotion bases, aqueous or non-aqueous liquid preparations, suspensions, emulsions, and the like. Examples of the ointment bases or the lotion bases include polyethylene glycol, propylene glycol, white vaseline, bleached bee wax, polyoxyethylene hydrogenated castor oil, glyceryl monostearate, stearyl alcohol, cetyl alcohol, lauromacrogol, sorbitan sesquioleate, and the like.
As the transmucosal agents such as an inhalation, a transnasal agent, and the like, those in the form of a solid, liquid, or semi-solid state are used, and can be prepared in accordance with a conventionally known method. For example, a known excipient, and also a pH adjusting agent, an antiseptic, a surfactant, a lubricant, a stabilizing agent, a thickening agent, or the like may be appropriately added thereto. For their administration, an appropriate device for inhalation or blowing can be used. For example, a compound may be administered alone or as a powder of formulated mixture, or as a solution or suspension in combination with a pharmaceutically acceptable carrier, using a conventionally known device or sprayer, such as a measured administration inhalation device, and the like. A dry powder inhaler or the like may be for single or multiple administration use, and a dry powder or a powder-containing capsule may be used. Alternatively, this may be in a form such as a pressurized aerosol spray which uses an appropriate propellant, for example, a suitable gas such as chlorofluoroalkane, hydrofluoroalkane, carbon dioxide, and the like, or other forms.
Generally, in the case of oral administration, the daily dose is from about 0.0001 to 100 mg/kg per body weight, administered in one portion or in 2 to 4 divided portions. In the case of intravenous administration, the daily dose is suitably administered from about 0.0001 to 10 mg/kg per body weight, once a day or two or more times a day. In addition, in the case of inhalation, the agent is administered at a dose from about 0.0001 to 1 mg/kg per body weight, once a day or two or more times a day. The dose is appropriately decided in response to the individual case by taking the symptoms, the age, the gender, and the like into consideration.
The compound of the formula (I) can be used in combination with various agents for treating or preventing the diseases for which the compound of the formula (I) of the present invention is considered to be effective. The combined preparations may be administered simultaneously, or separately and continuously, or at a desired time interval. The preparations to be co-administered may be a blend or may be prepared individually.
Hereinbelow, the preparation methods for the compound of the formula (I) are described in more detail with reference to the Examples. Further, the present invention is not intended to be limited to the compounds described in Examples below. In addition, the production processes for the starting compounds are shown in Preparation Examples. Further, the preparation methods for the compound of the formula (I) are not limited to the specific preparation methods in Examples presented below, but the compound of the formula (I) can be prepared by combinations of the preparation methods, or methods apparent to a skilled person in the art.
Moreover, the following abbreviations are used in some cases in Examples, Preparation Examples, and Tables to be Described Later.
PEx: Preparation Example No., Ex: Example No., Str: structural formula (a description of, for example, HCl, in the structural formula indicates that the compound is a hydrochloride, and a description of 2HCl indicates that the compound is dihydrochloride), rel: relative configuration (a description of rel under the PEx or Ex No. indicates that steric denotements in the adamantane skeletal portion in the structural formula described in the section of the Str represent relative configuration), Syn: Preparation Method (the numeral alone shows Example No. having the same preparation manner, and when P is prefixed before the number, the numeral shows Preparation Example No. having the same preparation manner), Dat: physicochemical data, NMR1: δ (ppm) 1H NMR in DMSO-d6, NMR2: δ (ppm) in 1H-NMR in CDCl3, NMR3: δ (ppm) 1H-NMR in D2O, FAB+:FAB-MS (positive ion), ESI+: ESI-MS (positive ion), ESI−: ESI-MS (negative ion), TEA: triethylamine, TFA: trifluoroacetic acid, THF: tetrahydrofuran, DMF: N,N-dimethylformamide, DME: dimethoxyethane, DMI: 1,3-dimethyl-2-imidazolidinone, MeOH: methanol, EtOH: ethanol, EtOAc: ethyl acetate, MeCN: acetonitrile, HOBt: 1-hydroxybenzotriazole, WSC: 3-ethyl-1-(3-dimethylaminopropyl)carbodiimide, DEAD: diethylazodicarboxylate, DIPEA: diisopropylethylamine, MCPBA: m-chloroperbenzoic acid, LAH: lithium aluminum hydride, Pd/C: palladium on carbon, TLC1: TLC analysis (condition: eluting solvent; MeOH/chloroform=1/9, silica gel plate (silica gel 60 F254, Merck)), TLC2: TLC analysis (condition: eluting solvent; hexane/EtOAc=1/1, amino silica gel plate (TLC plate (NH), FUJI SILYSIA)), TLC3: TLC analysis (condition: eluting solvent; EtOAc, amino silica gel plate (TLC plate (NH), FUJI SILYSIA)), HPLC: HPLC analysis, rt: retention time.
Further, there are descriptions of the retention time (HPLC:rt) in HPLC in the physicochemical data, in which the HPLC analysis conditions are as follows.
(Analysis Conditions)
Column: YMC-Pack ODS-AM (S-5 μm, 12 nm) (150×4.6 mm I.D.), Column temperature: 40° C., Detection method: UV (254 nm), Flow rate: 1 mL/min, Eluent A: acetonitrile, Eluent B: pH 3 buffer (phosphoric acid being added to a 0.05 M aqueous NaH2PO4 solution to adjust to pH 3)
To a solution of rel-[(1R,3S,5S)-5-({[(benzyloxy)carbonyl]amino}methyl)adamantan-2-yl]acetic acid (250 mg) in toluene (3 ml) were sequentially added TEA (127 μl) and diphenylphosphoryl azide (196 μl), followed by stirring at 80° C. for 1 hour. After leaving to be cooled to room temperature, to the mixed reaction liquid were sequentially added copper (I) iodide (69 mg) and tert-butanol (3 ml), followed by stirring at 80° C. for 1 hour. The mixed reaction liquid was diluted with EtOAc, and the organic layer was sequentially washed with water and saturated brine, and dried over anhydrous sodium sulfate. After the desiccant was removed, the solvent was evaporated under reduced pressure. The obtained residue was purified by silica gel flash column chromatography (hexane-EtOAc) to obtain 113 mg of benzyl tert-butyl[(1S,3R,5S)-tricyclo[3.3.1.13,7]decane-1,4-diyl bis(methylene)]bis rel-carbamate.
Under ice-cooling, to a suspension of 60% sodium hydride (oil dispersion, 25.5 mg) in THF (1 ml) was added dropwise triethyl phosphonoacetate (0.128 ml), followed by stirring for 10 minutes. To the mixed reaction liquid was added portionwise benzyl rel-{[(1S,3R,5S)-4-oxoadamantan-1-yl]methyl}carbamate (100 mg) at the same temperature, and the mixed reaction liquid was stirred at room temperature for 1 hour. To the mixed reaction liquid were added EtOAc and water, and the organic layer was collected by separation. The organic layer was sequentially washed with water and saturated brine, and dried over anhydrous sodium sulfate. After the desiccant was removed, the solvent was evaporated under reduced pressure. The obtained residue was purified by silica gel flash column chromatography (hexane-EtOAc) to obtain 120 mg of ethyl rel-(2E)-[(1R,3S,5R)-5-({[(benzyloxy)carbonyl]amino}methyl)tricyclo[3.3.1.13,7]dec-2-ylidene]acetate.
To a solution of tert-butyl rel-{(1R,2S,3S,5S)-5-[({2-[(3-bromobenzyl)amino]-5-cyanopyrimidine-4-yl}amino)methyl]adamantan-2-yl}carbamate (136 mg) in DME (2.7 ml) were added (3-aminomethylphenyl)boronic acid hydrochloride (89.8 mg), tetrakis(triphenylphosphine)palladium (0) (41.5 mg), sodium carbonate (101.6 mg), and water (0.34 ml), followed by stirring at 140° C. for 6 hours under a nitrogen air flow. To the mixed reaction liquid were added (3-aminomethylphenyl)boronic acid hydrochloride (89.8 mg) and a 2 M aqueous sodium carbonate solution (0.479 ml), followed by stirring at 140° C. for additional 4 hours. The mixed reaction liquid was diluted with EtOAc, and the organic layer was sequentially washed with water and saturated brine, and dried over anhydrous sodium sulfate. After the desiccant was removed, the solvent was evaporated under reduced pressure. The obtained residue was purified by amino silica gel flash column chromatography (chloroform-MeOH) to obtain 126.6 mg of tert-butyl rel-[(1R,2S,3S,5S)-5-({[2-({[3′-(aminomethyl)biphenyl-3-yl]methyl}amino)-5-cyanopyrimidin-4-yl]amino}methyl)adamantan-2-yl]carbamate.
To a solution of tert-butyl N-(5-cyano-2-{[2-(trifluoromethoxy)benzyl]amino}pyrimidin-4-yl)glycinate (616 mg) in dichloromethane (6.16 ml) was added trifluoroacetic acid (3.4 ml), followed by stirring at room temperature. After completion of the reaction, to the mixed reaction liquid was added diisopropyl ether. The precipitated solid was collected by filtration, washed with diisopropyl ether, and dried under reduced pressure to obtain 484 mg of N-(5-cyano-2-{[2-(trifluoromethoxy)benzyl]amino}pyrimidin-4-yl)glycine trifluoroacetate.
To a solution of methyl 1H-benzimidazole-5-carboxylate (8.5 g) in THF (85 ml) were added 3,4-dihydro-2H-pyran (5.3 ml) and (1S)-(+)-10-camphorsulfonic acid (1.1 g), followed by heating and refluxing for 24 hours. To the mixed reaction liquid were added 3,4-dihydro-2H-pyran (4.4 ml) and (1S)-(+)-10-camphorsulfonic acid (10.1 g), followed by heating and refluxing for additional 12 hours. The mixed reaction liquid was poured into a mixed liquid of EtOAc and water, and the organic layer was collected by separation. The organic layer was sequentially washed with water and saturated brine, and dried over anhydrous sodium sulfate. After the desiccant was removed, the solvent was evaporated under reduced pressure. The obtained residue was purified by silica gel flash column chromatography (hexane-EtOAc) to obtain a mixture (7.46 g) of methyl 1-(tetrahydro-2H-pyran-2-yl)-1H-benzimidazole-5-carboxylate and methyl 1-(tetrahydro-2H-pyran-2-yl)-1H-benzimidazole-6-carboxylate.
Under ice-cooling, to a solution of tert-butyl rel-[(1R,2S,3S,5S)-5-(aminomethyl)adamantan-2-yl]carbamate (200 mg) and TEA (0.12 ml) in dichloromethane (4 ml) was added benzyl chloroformate (0.11 ml), followed by stirring at room temperature for 4 hours. The reaction liquid was diluted with EtOAc, sequentially washed with 0.1 M hydrochloric acid, water, saturated aqueous sodium bicarbonate, water, and saturated brine, and dried over anhydrous sodium sulfate. After the desiccant was removed, the solvent was evaporated under reduced pressure, and the obtained residue was purified by silica gel column chromatography (hexane-EtOAc) to obtain 295.0 mg of benzyl rel-({(1S,3R,4S,5S)-4-[(tert-butoxycarbonyl)amino]adamantan-1-yl}methyl)carbamate.
Under ice-cooling, to a suspension of rel-1-[(1′R,3′S,5′S)-5′H-spiro[1,3-dioxolane-2,2′-tricyclo[3.3.1.13,7]decan]-5′-yl]methanamine (2.15 g) in THF (21.5 ml) were added dropwise benzyl chloroformate (1.92 ml) and a 1 M aqueous sodium hydroxide solution (13.5 ml). The mixed reaction liquid was warmed to room temperature, followed by stirring at room temperature for 3 hours. The mixed reaction liquid was diluted with EtOAc and then adjusted to pH 3 with an aqueous sodium hydrogen sulfate solution, and the organic layer was collected by separation. The organic layer was sequentially washed with water and saturated brine, and dried over anhydrous sodium sulfate. After the desiccant was removed, the solvent was evaporated under reduced pressure to obtain 2.66 g of benzyl rel-[(1′R,3′S,5′S)-5′H-spiro[1,3-dioxolane-2,2′-tricyclo[3.3.1.13,7]decan]-5′-ylmethyl]carbamate.
A suspension of {trans-3-[(tert-butoxycarbonyl)amino]cyclobutyl}methyl methanesulfonate (80.7 mg) and sodium azide (93.9 mg) in DMF (0.81 ml) and water (0.081 ml) was stirred at 120° C. for 40 minutes. The reaction liquid was cooled, then diluted with EtOAc, washed with water and saturated brine in this order, and dried over anhydrous sodium sulfate. After the desiccant was removed, the solvent was evaporated under reduced pressure, and the obtained residue was purified by silica gel column chromatography (hexane-EtOAc) to obtain 63.1 mg of tert-butyl[trans-3-(azidomethyl)cyclobutyl]carbamate.
To a solution of tert-butyl[trans-3-(azidomethyl)cyclobutyl]carbamate (270 mg) in MeOH (13.5 ml) was added 10% Pd/C (wetted with 50% water, 81 mg), followed by stirring at room temperature for 40 minutes at a normal pressure under a hydrogen atmosphere. The catalyst was separated by filtration through Celite and washed with MeOH, and then the filtrate was concentrated under reduced pressure. The obtained residue was purified by silica gel column chromatography (chloroform-MeOH-concentrated aqueous ammonia) to obtain 120.5 mg of tert-butyl[trans-3-(aminomethyl)cyclobutyl]carbamate.
To a solution of N-(5-cyano-2-{[2-(trifluoromethoxy)benzyl]amino}pyrimidin-4-yl)glycine trifluoroacetate (30 mg) in DMF (0.9 ml) were sequentially added tert-butyl (2-aminoethyl)carbamate (25.0 mg), HOBt (9.3 mg), and WSC (24.2 mg), followed by stirring at room temperature. After completion of the reaction, the mixed reaction liquid was diluted with EtOAc, and the organic layer was sequentially washed with water and saturated brine, and dried over anhydrous sodium sulfate. After the desiccant was removed, the solvent was evaporated under reduced pressure. The obtained residue was purified by preparative silica gel thin layer chromatography (chloroform-MeOH) to obtain 10 mg of tert-butyl (2-{[N-(5-cyano-2-{[2-(trifluoromethoxy)benzyl]amino}pyrimidin-4-yl)glycyl]amino}ethyl)carbamate.
Under ice-cooling, to a solution of benzyl rel-[(1R,2S,3S,5S)-5-(aminomethyl)adamantan-2-yl]carbamate and pyridine (1.4 ml) in dichloromethane (50 ml) were added a solution of trifluoroacetic anhydride (2.5 ml) in dichloromethane (20 ml), followed by stirring at the same temperature for 30 minutes. Pyridine (0.128 ml) and trifluoroacetic anhydride (0.225 ml) were further added thereto, followed by stirring for 30 minutes under ice-cooling. Under ice-cooling, to the mixed reaction liquid was added water, followed by stirring and then dilution with EtOAc, and the organic layer was sequentially washed with water and saturated brine, and dried over anhydrous sodium sulfate. After the desiccant was removed, the solvent was evaporated under reduced pressure to obtain 7.0 g of benzyl rel-[(1R,2S,3S,5S)-5-{[(trifluoroacetyl)amino]methyl}adamantan-2-yl]carbamate.
To a solution of rel-(1R,3S,5R,7S)-4-{[benzyloxy)carbonyl]amino}adamantane-1-carboxylic acid (12 g) in dichloromethane (120 ml) were added oxalyl chloride (4.8 ml), followed by stirring at room temperature. After completion of the reaction, the reaction mixture was concentrated under reduced pressure, and toluene was added thereto, followed by additional concentration under reduced pressure. The obtained residue was dissolved in 1,4-dioxane (12 ml), and added dropwise to 28% aqueous ammonia (110 g) under ice-cooling. The mixed reaction liquid was extracted with EtOAc, and the organic layer was washed with water three times and with saturated brine, and dried over anhydrous sodium sulfate. After the desiccant was removed, the solvent was evaporated under reduced pressure. To the obtained residue was added MeOH to precipitate the solid, which was collected by filtration. The filtrate was concentrated under reduced pressure, and MeOH was used again to precipitate the solid, which was collected by filtration. The filtrate was concentrated under reduced pressure, and MeOH was used several times to precipitate the solid, which was collected by filtration. The obtained solid was dried under reduced pressure to obtain benzyl rel-[(1R,2R,3S,5S)-5-carbamoyladamantan-2-yl]carbamate (2.9 g). The filtrate was concentrated under reduced pressure and the obtained residue was purified by silica gel flash column chromatography (hexane-EtOAc) to obtain 1.9 g of benzyl rel-[(1R,2S,3S,5S)-5-carbamoyladamantan-2-yl]carbamate.
To a mixed solution of tert-butyl rel-[(1R,2S,3S,5S)-5-{[(trifluoroacetyl)amino]methyl}adamantan-2-yl]carbamate (4.6 g) in MeOH (46 ml) and water (23 ml) was added potassium carbonate (16.9 g), followed by stirring at room temperature. After completion of the reaction, the mixed reaction liquid was diluted with EtOAc, washed with saturated brine, and dried over anhydrous sodium sulfate. After the desiccant was removed, the solvent was evaporated under reduced pressure to obtain 3.78 g of tert-butyl rel-[(1R,2S,3S,5S)-5-(aminomethyl)adamantan-2-yl]carbamate.
Under ice-cooling, to a solution of benzyl rel-[(1R,2S,3S,5S)-5-carbamoyladamantan-2-yl]carbamate (500 mg) in THF (5.0 ml) was added dropwise a 1.17 M solution of a borane-tetrahydrofuran complex in THF (3.9 ml) under a nitrogen air flow, followed by heating and refluxing for 3 hours. The mixed reaction liquid was ice-cooled, and then water was carefully added dropwise thereinto. Then, the liquid was poured into an aqueous dichloromethane-potassium carbonate solution under stirring. The organic layer was collected by separation, and further extracted with dichloromethane twice. The obtained organic layer was combined and dried over anhydrous sodium sulfate. After the desiccant was removed, the solvent was evaporated under reduced pressure to obtain 530 mg of benzyl rel-[(1R,2S,3S,5S)-5-(aminomethyl)adamantan-2-yl]carbamate.
Under ice-cooling, to a solution of di-tert-butyl iminodicarboxylate (1.88 g) in DMF (28 ml) was added potassium tert-butoxide (970 mg) in small portions, followed by stirring at room temperature for 1 hour. To the reaction mixture was added dropwise a solution of 3-(bromomethyl)-4-chlorophenyl acetate (1.90 g) in DMF (10 ml) under ice-cooling, followed by stirring at room temperature for 2 hours. To the reaction mixture was added water, followed by extraction with EtOAc. The organic layer was sequentially washed with water and saturated brine, and dried over anhydrous magnesium sulfate. After the desiccant was removed, the solvent was evaporated under reduced pressure. The obtained residue was purified by silica gel column chromatography (hexane-EtOAc) to obtain 2.79 g of 3-{[bis(tert-butoxycarbonyl)amino]methyl}-4-chlorophenyl acetate.
Under an argon atmosphere, to a solution of [2-(benzyloxy)phenyl]methanol (20.2 g) in chloroform (160 ml) was slowly added a solution of thionyl chloride (13.8 ml) in chloroform (40 ml) at room temperature. After stirring at room temperature for 90 minutes, volatile substances were evaporated under reduced pressure to obtain 1-(benzyloxy)-2-(chloromethyl)benzene. Then, under an argon atmosphere, to a solution of di-tert-butyl iminodicarboxylate (41.0 g) in DMF (500 ml) was added potassium tert-butoxide (21.2 g) at room temperature. After stirring at the same temperature for 70 minutes, a solution of 1-(benzyloxy)-2-(chloromethyl)benzene in DMF (60 ml) was added thereto. After stirring at the same temperature for 15 hours, water was added thereto, followed by stirring for additional 90 minutes. The precipitate was collected by filtration, washed with water, and then dried under reduced pressure. The obtained crude product was purified by silica gel column chromatography (chloroform) to obtain 34.5 g of di-tert-butyl[2-(benzyloxy)benzyl]imidodicarbonate.
To a solution of rel-4-({[(1S,3R,4S,5S)-4-aminoadamantan-1-yl]methyl}amino)-2-{[2-(trifluoromethoxy)benzyl]amino}pyrimidine-5-carbonitrile (30 mg) in DMF (0.6 ml) were added DIPEA (22 μl) and ethylbromo acetate (5.8 μl), followed by stirring at 60° C. After completion of the reaction, the mixed reaction liquid was diluted with EtOAc, and the organic layer was sequentially washed with water and saturated brine, and dried over anhydrous sodium sulfate. After the desiccant was removed, the solvent was evaporated under reduced pressure. The obtained residue was purified by preparative silica gel thin layer chromatography (chloroform-MeOH) to obtain ethyl rel-N-[(1R,2S,3S,5S)-5-{[(5-cyano-2-{[2-(trifluoromethoxy)benzyl]amino}pyrimidin-4-yl)amino]methyl}adamantan-2-yl]glycinate (26.1 mg).
To a solution of di-tert-butyl (2-hydroxybenzyl)imidodicarbonate (500 mg) in DMF (5.0 ml) were added 2-bromoacetamide (320 mg), potassium carbonate (641 mg), and potassium iodide (385 mg), followed by stirring at 80° C. for 3 hours. After leaving to be cooled to room temperature, water was added thereto, and the precipitated product was collected by filtration to obtain 546 mg of di-tert-butyl[2-(2-amino-2-oxoethoxy)benzyl]imidodicarbonate.
To a mixed solution of tert-butyl (5-formyl-2-methoxybenzyl)carbamate (1.0 g) in THF (3.0 ml) and EtOH (6.0 ml) was added sodium borohydride (192.5 mg), followed by stirring at room temperature. After completion of the reaction, to the mixed reaction liquid was added water, followed by extraction with EtOAc, and the organic layer was dried over anhydrous sodium sulfate. After the desiccant was removed, the solvent was evaporated under reduced pressure. The obtained residue was purified by silica gel flash column chromatography (hexane-EtOAc) to obtain 1.09 g of tert-butyl[5-(hydroxymethyl)-2-methoxybenzyl]carbamate.
To a solution of 4-chloro-2-(methylsulfanyl)pyrimidine-5-carbonitrile (2.2 g) in 1,3-dimethylimidazolidin-2-one were added DIPEA (4.13 ml) and tert-butyl rel-[(1R,2S,3S,5S)-5-(aminomethyl)adamantan-2-yl]carbamate (3.99 g), followed by stirring at room temperature. After completion of the reaction, the mixed reaction liquid was diluted with EtOAc, and the organic layer was sequentially washed with water and saturated brine, and dried over anhydrous sodium sulfate. After the desiccant was removed, the solvent was evaporated under reduced pressure. The obtained residue was purified by silica gel flash column chromatography (hexane-EtOAc) to obtain 4.76 g of tert-butyl rel-[(1R,2S,3S,5S)-5-({[5-cyano-2-(methyl sulfanyl)pyrimidin-4-yl]amino}methyl)adamantan-2-yl]carbamate.
Under ice-cooling, to a solution of 2,4-dichloropyrimidine-5-carbonitrile (1.00 g) in DMF (15 ml) were added dropwise a solution of 2-(methylthio)benzylamine (881 mg) in DMF (5 ml) and DIPEA (1.2 ml), followed by stirring at the same temperature for 1 hour. A solution of 2-(methylthio)benzylamine (44 mg) in DMF (2 ml) was added thereto, followed by stirring at room temperature for additional 1 hour. To the reaction mixture were added EtOAc and water, followed by liquid separation. The organic layer was sequentially washed with water and saturated brine, and dried over anhydrous magnesium sulfate. After the desiccant was removed, the solvent was evaporated under reduced pressure, and the obtained residue was purified by silica gel column chromatography (chloroform) to obtain 709 mg of 4-chloro-2-{[2-(methylsulfanyl)benzyl]amino}pyrimidine-5-carbonitrile.
To a solution of 2,4-dichloro-5-(trifluoromethyl)pyrimidine (300 mg) in DMF (6.0 ml) which had been cooled in an ice-brine bath were added DIPEA (252.9 μl) and 1-[2-(trifluoromethoxy)phenyl]methanamine (277.5 mg), followed by stirring at −20° C. After completion of the reaction, the mixed reaction liquid was diluted with EtOAc, and the organic layer was sequentially washed with water and saturated brine, and dried over anhydrous sodium sulfate. After the desiccant was removed, the solvent was evaporated under reduced pressure to obtain 507 mg of a mixture of 4-chloro-N-[2-(trifluoromethoxy)benzyl]-5-(trifluoromethyl)pyrimidin-2-amine and 2-chloro-N-[2-(trifluoromethoxy)benzyl]-5-(trifluoromethyl)pyrimidin-4-amine.
To a solution of a mixture (90 mg) of 4-chloro-N-[2-(trifluoromethoxy)benzyl]-5-(trifluoromethyl)pyrimidin-2-amine and 2-chloro-N-[2-(trifluoromethoxy)benzyl]-5-(trifluoromethyl)pyrimidin-4-amine in DMF (1.0 ml) were added DIPEA (84.3 μl) and benzyl rel-[(1R,2S,3S,5S)-5-(aminomethyl)adamantan-2-yl]carbamate (79.9 mg), followed by stirring at room temperature. After completion of the reaction, the mixed reaction liquid was diluted with EtOAc, and the organic layer was sequentially washed with water and saturated brine, and dried over anhydrous sodium sulfate. After the desiccant was removed, the solvent was evaporated under reduced pressure. The obtained residue was purified by silica gel flash column chromatography (hexane-EtOAc) to obtain a crude product of benzyl rel-[(1R,2S,3S,5S)-5-({[2-{[2-(trifluoromethoxy)benzyl]amino}-5-(trifluoromethyl)pyrimidin-4-yl]amino}methyl)adamantan-2-yl]carbamate and a crude product of benzyl rel-[(1R,2S,3S,5S)-5-({[4-{[2-(trifluoromethoxy)benzyl]amino}-5-(trifluoromethyl)pyrimidin-2-yl]amino}methyl)adamantan-2-yl]carbamate. Each of the crude products was further purified by silica gel flash column chromatography (chloroform-MeOH) to obtain benzyl rel-[(1R,2S,3S,5S)-5-({[2-{[2-(trifluoromethoxy)benzyl]amino}-5-(trifluoromethyl)pyrimidin-4-yl]amino}methyl)adamantan-2-yl]carbamate (100 mg) and benzyl rel-[(1R,2S,3S,5S)-5-({[4-{[2-(trifluoromethoxy)benzyl]amino}-5-(trifluoromethyl)pyrimidin-2-yl]amino}methyl)adamantan-2-yl]carbamate (50 mg).
To a solution of benzyl rel-[(1R,2S,3S,5S)-5-({[5-cyano-2-(methylsulfinyl)pyrimidin-4-yl]amino}methyl)adamantan-2-yl]carbamate (50 mg) in 1,3-dimethylimidazolidin-2-one (1.0 ml) were added 3-bromoaniline (0.114 ml) and a 4 M hydrogen chloride dioxane solution (2.6 μl), followed by stirring at 100° C. for 3 hours. After leaving to be cooled to room temperature, to the mixed reaction liquid was added water, and the precipitated solid was collected by filtration, washed with water and hexane, and then dried under reduced pressure to obtain 46 mg of benzyl rel-{(1R,2S,3S,5S)-5-[({2-[(3-bromophenyl)amino]-5-cyanopyrimidine-4-yl}amino)methyl]adamantan-2-yl}carbamate.
To a solution of ethyl rel-[(1R,3S,5S)-5-({[(benzyloxy)carbonyl]amino}methyl)adamantan-2-yl]acetate (300 mg) in MeOH (6.0 ml) was added a 4 M aqueous lithium hydroxide solution (1.2 ml), followed by stirring at 60° C. for 3 hours. The mixed reaction liquid was diluted with EtOAc and then an aqueous potassium hydrogen sulfate solution was added to adjust to pH 2, and the organic layer was collected by separation. The organic layer was sequentially washed with water and saturated brine, and dried over anhydrous sodium sulfate. After the desiccant was removed, the solvent was evaporated under reduced pressure to obtain 264.6 mg of rel-[(1R,3S,5S)-5-({[(benzyloxy)carbonyl]amino}methyl)adamantan-2-yl]acetic acid.
To a mixed solution of methyl rel-(1R,3S,5R,7S)-4-{[(benzyloxy)carbonyl]amino}adamantane-1-carboxylate (15 g) in 1,4-dioxane (75 ml) and MeOH (75 ml) were added a 1 M aqueous sodium hydroxide solution (87.4 ml), followed by stirring at 60° C. for 4 hours. The mixed reaction liquid was left to be cooled to room temperature, then adjusted to pH 4 with a 10% aqueous potassium hydrogen sulfate solution, and extracted with EtOAc. The obtained organic layer was washed with saturated brine once and dried over anhydrous magnesium sulfate. After the desiccant was removed, the solvent was evaporated under reduced pressure to obtain 12 g of rel-(1R,3S,5R,7S)-4-{[(benzyloxy)carbonyl]amino}adamantane-1-carboxylic acid.
Under a nitrogen atmosphere, to a suspension of lithium aluminum hydride (1.2 g) in THF (100 ml) was added dropwise a solution of a mixture (7.0 g) of methyl 1-(tetrahydro-2H-pyran-2-yl)-1H-benzimidazole-5-carboxylate and methyl 1-(tetrahydro-2H-pyran-2-yl)-1H-benzimidazole-6-carboxylate in THF (100 ml) at −10° C. or lower, followed by stirring for 1 hour under ice-cooling. Lithium aluminum hydride (0.8 g) was added in divided portions thereto, followed by stirring for additional 30 minutes under ice-cooling. At the same temperature, water (6.0 ml), a 15% aqueous sodium hydroxide solution (6.0 ml), and water (3.0 ml) were sequentially added thereto, followed by stirring at room temperature for 30 minutes. The insoluble materials were removed by filtration through Celite and the filtrate was concentrated under reduced pressure to obtain 5.48 g of a mixture of [1-(tetrahydro-2H-pyran-2-yl)-1H-benzimidazol-5-yl]methanol and [1-(tetrahydro-2H-pyran-2-yl)-1H-benzimidazol-6-yl]methanol.
Under ice-cooling, to a solution of ethyl rel-(2E)-[(1R,3S,5R)-5-({[(benzyloxy)carbonyl]amino}methyl)tricyclo[3.1.13,7]dec-2-ylidene]acetate (350 mg) in MeOH (6.0 ml) was added nickel (II) chloride (23.7 mg) under a nitrogen atmosphere, and sodium borohydride was added portionwise thereto, followed by stirring at the same temperature for 1 hour and at room temperature for 3 hours. To the mixed reaction liquid was added water, followed by extraction with EtOAc. The organic layer was sequentially washed with water and saturated brine, and dried over anhydrous sodium sulfate. After the desiccant was removed, the solvent was evaporated under reduced pressure to obtain 310 mg of ethyl rel-[(1R,3S,5S)-5-({[(benzyloxy)carbonyl]amino}methyl)adamantan-2-yl]acetate.
To a solution of methyl rel-(1R,3S,5R,7S)-4-oxoadamantane-1-carboxylate (500 mg) in dichloromethane (7.5 ml) were sequentially added benzyl amine (0.262 ml) and sodium triacetoxyborohydride (763 mg), followed by stirring at room temperature for 2 hours. To the mixed reaction liquid was added saturated aqueous sodium bicarbonate, followed by stirring and then extraction with dichloromethane. The organic layer was collected by separation. The obtained organic layer was washed with saturated brine and dried over anhydrous magnesium sulfate. After the desiccant was removed, the solvent was evaporated under reduced pressure. The obtained residue was purified by silica gel flash column chromatography (chloroform-MeOH) to obtain 757 mg of methyl rel-(1R,3S,5R,7S)-4-(benzylamino)adamantane-1-carboxylate.
To a solution of benzyl rel-{[(1S,3R,4S,5S)-4-aminoadamantan-1-yl]methyl}carbamate (760 mg) in dichloromethane (22.8 ml) were added 4-{[tert-butyl(dimethyl)silyl]oxy}cyclohexanone (1.22 ml) and sodium triacetoxyborohydride (1.02 g), followed by stirring at room temperature for 4 hours. To the mixed reaction liquid was added saturated aqueous sodium bicarbonate, followed by extraction with EtOAc. The organic layer was sequentially washed with water and saturated brine, and dried over anhydrous sodium sulfate. The desiccant was removed, the solvent was evaporated under reduced pressure, and the obtained residue was purified by amino silica gel column chromatography (hexane-EtOAc) to first elute 674.8 mg of benzyl rel-({(1R,3S,4R,5R)-4-[(cis-4-{[tert-butyl(dimethyl)silyl]oxy}cyclohexyl)amino]adamantan-1-yl}methyl)carbamate and then elute 435.8 mg of benzyl rel-({(1R,3S,4R,5R)-4-[(trans-4-{[tert-butyl(dimethyl)silyl]oxy}cyclohexyl)amino]adamantan-1-yl}methyl)carbamate.
The steric configuration of the obtained product was determined by using the compound (benzyl rel-({(1R,3S,4R,5R)-4-[(trans-4-{[tert-butyl (dimethyl)silyl]oxy}cyclohexyl)amino]adamantan-1-yl}methyl)carbamate) eluted later in amino silica gel column chromatography as a starting material to provide the rel-trans-4-{[(1R,2S,3S,5S)-5-(aminomethyl)adamantan-2-yl]amino}cyclohexanol obtained in Preparation Example 134, which is then used for Example 45, and by confirming that the HPLC retention time (15.1 min) of the obtained product coincided with that in Example 42 (trans-alcohol product).
Under ice-cooling, to a solution of tert-butyl rel-[(1R,2S,3S,5S)-5-({[5-cyano-2-(methylsulfanyl)pyrimidin-4-yl]amino}methyl)adamantan-2-yl]carbamate (4.7 g) in dichloromethane (50 ml) was added 75% MCPBA (contains water) (2.77 g), followed by stirring at the same temperature. After completion of the reaction, the mixed reaction liquid was diluted with EtOAc, and the organic layer was sequentially washed with saturated aqueous sodium bicarbonate, water, and saturated brine, and dried over anhydrous sodium sulfate. After the desiccant was removed, the solvent was evaporated under reduced pressure. The obtained residue was purified by silica gel flash column chromatography (chloroform-MeOH) to obtain 5.02 g of tert-butyl rel-[(1R,2S,3S,5S)-5-({[5-cyano-2-(methylsulfinyl)pyrimidin-4-yl]amino}methyl)adamantan-2-yl]carbamate.
Under ice-cooling, to a solution of tert-butyl rel-[(1R,2S,3S,5S)-5-{[(5-cyano-2-{[2-(methylsulfanyl)benzyl]amino}pyrimidin-4-yl)amino]methyl}adamantan-2-yl]carbamate (147 mg) in dichloromethane (5.0 ml) was added 75% MCPBA (contains water, 69.6 mg), followed by stirring at the same temperature for 1 hour. To the mixed reaction liquid was added saturated aqueous sodium bicarbonate, followed by extraction with EtOAc. The organic layer was sequentially washed with water and saturated brine, and dried over anhydrous sodium sulfate. After the desiccant was removed, the solvent was evaporated under reduced pressure. The obtained residue was purified by silica gel flash column chromatography (starting with hexane-EtOAc and changing to chloroform-MeOH in the middle of the process) to obtain tert-butyl rel-[(1R,2S,3S,5S)-5-{[(5-cyano-2-{[2-(methylsulfinyl)benzyl]amino}pyrimidin-4-yl)amino]methyl}adamantan-2-yl]carbamate (123.9 mg) and tert-butyl rel-[(1R,2S,3S,5S)-5-{[(5-cyano-2-{[2-(methylsulfonyl)benzyl]amino}pyrimidin-4-yl)amino]methyl}adamantan-2-yl]carbamate (28.1 mg).
Under ice-cooling, to a suspension of LAH (88 mg) in THF (20 ml) was added tert-butyl 3-cyano-8-azabicyclo[3.2.1]octane-8-carboxylate (550 mg), followed by stirring at room temperature for 4 hours. Under ice-cooling, water was added thereto, followed by extraction with EtOAc, and the organic layer was dried over anhydrous sodium sulfate. After the desiccant was removed, the solvent was evaporated under reduced pressure to obtain 600 mg of tert-butyl 3-(aminomethyl)-8-azabicyclo[3.2.1]octane-8-carboxylate.
To a solution of di-tert-butyl{2-[2-(methoxymethoxy)ethoxy]benzyl}imidodicarbonate (288 mg) in methanol (1.4 ml) was added a 4 M hydrogen chloride dioxane solution (3.5 ml), followed by stirring at room temperature for 2 hours. The solvent was evaporated under reduced pressure to obtain 140 mg of 2-[2-(aminomethyl)phenoxy]ethanol hydrochloride.
Under ice-cooling, to a solution of benzyl rel-({(1S,3R,4S,5S)-4-[(tert-butoxycarbonyl)amino]adamantan-1-yl}methyl)carbamate (295.0 mg) in dichloromethane (3.54 ml) was added trifluoroacetic acid (3.54 ml), followed by stirring at room temperature for 2 hours. The reaction liquid was concentrated under reduced pressure, and then the residue was alkalified by the addition of an aqueous potassium carbonate solution and then extracted with EtOAc. The organic layer was sequentially washed with water and saturated brine, and dried over anhydrous sodium sulfate. After the desiccant was removed, the solvent was evaporated under reduced pressure to obtain 242.8 mg of benzyl rel-{[(1S,3R,4S,5S)-4-aminoadamantan-1-yl]methyl}carbamate.
To a solution of di-tert-butyl[2-(2-methoxyethoxy)benzyl]imidodicarbonate (341 mg) in 1,4-dioxane (1.7 ml) was added a 4 M hydrogen chloride dioxane solution (3.5 ml) at room temperature, followed by stirring for 2 hours. The solvent was evaporated under reduced pressure to obtain 155 mg of 1-[2-(2-methoxyethoxy)phenyl]methanamine hydrochloride.
To a solution of tert-butyl (2-{4-[(methylsulfonyl)amino]phenyl}ethyl)carbamate (900 mg) in dichloromethane (18 ml) was added trifluoroacetic acid (2.89 ml), followed by stirring at room temperature overnight. The mixed reaction liquid was concentrated under reduced pressure, and toluene was added to the residue, followed by further concentration under reduced pressure. To the obtained residue was added diethyl ether, the precipitated solid was collected by filtration, washed with diethyl ether, and then dried under reduced pressure. The obtained solid was suspended in EtOH, alkalified by the addition of a 1 M aqueous sodium hydroxide solution, then adjusted to pH 7 with 1 M hydrochloric acid, and extracted with chloroform. The organic layer was combined and dried over anhydrous sodium sulfate. After the desiccant was removed, the solvent was evaporated under reduced pressure to obtain 95 mg of N-[4-(2-aminoethyl)phenyl]methanesulfonamide.
Under ice-cooling, to a solution of benzyl rel-[(1R,2S,3S,5S)-5-{[(trifluoroacetyl)amino]methyl}adamantan-2-yl]carbamate (7.0 g) in EtOH (175 ml) were sequentially added di-tert-butyl dicarbonate (5.58 g), 10% Pd/C (wetted with 50% water, 7.0 g), and cyclohexa-1,4-diene (15.9 ml), followed by stirring at room temperature for 1 hour. The catalyst was removed by filtration, and then the filtrate was concentrated under reduced pressure. The obtained residue was purified by silica gel flash column chromatography (hexane-EtOAc) to obtain 4.67 g of tert-butyl rel-[(1R,2S,3S,5S)-5-{[(trifluoroacetyl)amino]methyl}adamantan-2-yl]carbamate.
To a solution of benzyl rel-({(1R,3S,4R,5R)-4-[(trans-4-hydroxycyclohexyl)amino]adamantan-1-yl}methyl)carbamate (380 mg) in MeOH (11.4 ml) was added 10% Pd/C (wetted with 50% water, 76 mg), followed by stirring at 35° C. for 2.5 hours at a normal pressure under a hydrogen atmosphere. The catalyst was separated by filtration through Celite and washed with MeOH, and then the filtrate was concentrated under reduced pressure to obtain 263.8 mg of rel-trans-4-{[(1R,2S,3S,5S)-5-(aminomethyl)adamantan-2-yl]amino}cyclohexanol.
To a solution of 3-{[bis(tert-butoxycarbonyl)amino]methyl}-4-chlorophenyl acetate (2.79 g) in methanol (56 ml) was added potassium carbonate (1.45 g), followed by stirring at room temperature for 1 hour. To the reaction mixture was added saturated aqueous ammonium chloride solution, and the precipitate was collected by filtration to obtain 2.15 g of di-tert-butyl (2-chloro-5-hydroxybenzyl)imidodicarbonate.
To a mixed solution of rel-4-{[(1′R,3′S,5′S)-5′H-spiro[1,3-dioxolane-2,2′-tricyclo[3.3.1.13,7]decan]-5′-ylmethyl]amino}-2-{[2-(trifluoromethoxy)benzyl]amino}pyrimidine-5-carbonitrile (385 mg) in THF (23.1 ml) and water (30.8 ml) was added p-toluenesulfonic acid monohydrate (1.42 g), followed by stirring at room temperature overnight. The mixed reaction liquid was concentrated under reduced pressure, and the residue was alkalified by the addition of saturated aqueous sodium bicarbonate, followed by extraction with EtOAc. The organic layer was washed with saturated brine and dried over anhydrous magnesium sulfate. After the desiccant was removed, the solvent was evaporated under reduced pressure. The obtained residue was purified by silica gel flash column chromatography (hexane-EtOAc) to obtain 300 mg of rel-4-({[(1S,3R,5S)-4-oxoadamantan-1-yl]methyl}amino)-2-{[2-(trifluoromethoxy)benzyl]amino}pyrimidine-5-carbonitrile.
To a mixed solution of benzyl rel-[(1′R,3′S,5′S)-5′H-spiro[1,3-dioxolane-2,2′-tricyclo[3.3.1.13,7]decan]-5′-ylmethyl]carbamate (2.5 g) in THF (25 ml) and water (25 ml) was added p-toluenesulfonic acid monohydrate (6.65 g), followed by stirring at room temperature overnight. The mixed reaction liquid was concentrated under reduced pressure, and the residue was alkalified by the addition of saturated aqueous sodium bicarbonate, followed by extraction with EtOAc. The obtained organic layer was washed with saturated brine and dried over anhydrous magnesium sulfate. After the desiccant was removed, the solvent was evaporated under reduced pressure to obtain 2.26 g of benzyl rel-{[(1S,3R,5S)-4-oxoadamantan-1-yl]methyl}carbamate.
To a solution of benzyl rel-({(1R,3S,4R,5R)-4-[(trans-4-{[tert-butyl(dimethyl)silyl]oxy}cyclohexyl)amino]adamantan-1-yl}methyl)carbamate (446 mg) in THF (8.92 ml) was added a solution (2.54 ml) of 1 M tetrabutylammonium fluoride in THF, followed by stirring at 70° C. for 5.5 hours. The solvent was evaporated under reduced pressure, and to the residue was added water, followed by extraction with chloroform. The organic layer was washed with saturated brine and dried over anhydrous sodium sulfate, the desiccant was then removed, and the solvent was evaporated under reduced pressure. The obtained residue was purified by silica gel column chromatography (starting with chloroform-MeOH and changing to chloroform-MeOH-concentrated aqueous ammonia in the middle of the process) to obtain 385.1 mg of benzyl rel-({(1R,3S,4R,5R)-4-[(trans-4-hydroxycyclohexyl)amino]adamantan-1-yl}methyl)carbamate.
To a mixed solution of a mixture (3.99 g) of 2-{[1-(tetrahydro-2H-pyran-2-yl)-1H-benzimidazol-5-yl]methyl}-1H-isoindole-1,3(2H)-dione and 2-{[1-(tetrahydro-2H-pyran-2-yl)-1H-benzimidazol-6-yl]methyl}-1H-isoindole-1,3(2H)-dione in EtOH (79.8 ml) and THF (79.8 ml) was added hydrazine monohydrate (2.14 ml), followed by heating and refluxing. After completion of the reaction, the insoluble materials were removed by filtration and the filtrate was concentrated under reduced pressure. The obtained residue was diluted with dichloromethane, washed with a 1 M aqueous sodium hydroxide solution, and dried over anhydrous magnesium sulfate. After the desiccant was removed, the solvent was evaporated under reduced pressure. The obtained residue was purified by amino silica gel flash column chromatography (chloroform-MeOH) and then purified by silica gel flash column chromatography (chloroform-MeOH) to obtain 0.42 g of a mixture of 1-[1-(tetrahydro-2H-pyran-2-yl)-1H-benzimidazol-5-yl]methanamine and 1-[1-(tetrahydro-2H-pyran-2-yl)-1H-benzimidazol-6-yl]methanamine.
To a mixed solution of methyl rel-(1R,3S,5R,7S)-4-(benzylamino)adamantane-1-carboxylate (720 mg) in EtOH (7.2 ml) and water (0.72 ml) were added 10% Pd/C (wetted with 50% water, 144 mg) and ammonium formate (455 mg), followed by heating and refluxing for 30 minutes. The mixed reaction liquid was left to be cooled to room temperature, then the catalyst was separated by filtration through Celite, and the filtrate was concentrated under reduced pressure to obtain 482 mg of methyl rel-(1R,3S,5R,7S)-4-aminoadamantane-1-carboxylate.
To a solution of di-tert-butyl[2-(benzyloxy)benzyl]imidodicarbonate (34.5 g) in methanol (170 ml) and THF (170 ml) was added 10% Pd/C (3.5 g), followed by stirring for 14 hours at a normal pressure under a hydrogen atmosphere. The reaction mixture was filtered and the filtrate was concentrated under reduced pressure to obtain 27.0 g of di-tert-butyl (2-hydroxybenzyl)imidodicarbonate.
To a solution of 4-chloro-3-methylphenyl acetate (2.06 g) in carbon tetrachloride (20.6 ml) were added N-bromosuccinimide (1.99 g) and 2,2′-azobis(isobutyronitrile) (366 mg), followed by heating and refluxing for 1 hour. To the reaction liquid was added water, followed by extraction with EtOAc, and the organic layer was washed with saturated aqueous sodium chloride solution, and then dried over anhydrous magnesium sulfate. After the desiccant was removed, the solvent was evaporated under reduced pressure, and the residue was purified by silica gel column chromatography (hexane-EtOAc) to obtain 1.87 g of 3-(bromomethyl)-4-chlorophenyl acetate.
To a solution of tert-butyl (2-methoxybenzyl)carbamate (25.0 g) in MeCN (200 ml) was added N-bromosuccinimide (19.7 g), followed by stirring at room temperature overnight. To the mixed reaction liquid was added N-bromosuccinimide (10.0 g), followed by additionally stirring at room temperature for 8 hours. The mixed reaction liquid was concentrated under reduced pressure, and then the residue was diluted with EtOAc, sequentially washed with water and saturated brine, and dried over anhydrous sodium sulfate. After the desiccant was removed, the solvent was evaporated under reduced pressure. The obtained residue was purified by silica gel flash column chromatography (toluene) to obtain 9.55 g of tert-butyl (5-bromo-2-methoxybenzyl)carbamate.
To a solution of tert-butyl (5-bromo-2-methoxy benzyl)carbamate (5.0 g) in DMF (50 ml) were added bis(triphenylphosphine)palladium (II) dichloride (222 mg), triphenyl phosphine (83 mg), and sodium hydrogen carbonate (1.61 g), followed by heating and stirring at 110° C. at a normal pressure under a carbon monoxide atmosphere. The mixed reaction liquid was diluted with EtOAc, sequentially washed with water, an aqueous sodium carbonate solution, and saturated brine, and dried over anhydrous sodium sulfate. After the desiccant was removed, the solvent was evaporated under reduced pressure. The obtained residue was purified by silica gel flash column chromatography (hexane-EtOAc) to obtain 1.69 g of tert-butyl (5-formyl-2-methoxybenzyl)carbamate.
Under ice-cooling, to a solution of a mixture (2.0 g) of [1-(tetrahydro-2H-pyran-2-yl)-1H-benzimidazol-5-yl]methanol and [1-(tetrahydro-2H-pyran-2-yl)-1H-benzimidazol-6-yl]methanol in toluene (40 ml) were added succinimide (1.52 g), triphenyl phosphine (2.71 g), and DEAD (1.62 ml), followed by stirring at the same temperature for 3 hours. The mixed reaction liquid was diluted with EtOAc, sequentially washed with water and saturated brine, and dried over anhydrous sodium sulfate. After the desiccant was removed, the solvent was evaporated under reduced pressure, and the obtained residue was purified by silica gel flash column chromatography (chloroform-MeOH) to obtain 4.12 g of a mixture of 2-{[1-(tetrahydro-2H-pyran-2-yl)-1H-benzimidazol-5-yl]methyl}-1H-isoindole-1,3(2H)-dione and 2-{[1-(tetrahydro-2H-pyran-2-yl)-1H-benzimidazol-6-yl]methyl}-1H-isoindole-1,3(2H)-dione.
Under ice-cooling, to a solution of tert-butyl[trans-3-(hydroxymethyl)cyclobutyl]carbamate (60 mg) and TEA (0.066 ml) in dichloromethane (4 ml) was added methanesulfonyl chloride (0.035 ml), followed by stirring at the same temperature for 30 minutes. The reaction liquid was diluted with EtOAc, washed with water and saturated brine in this order, and dried over anhydrous sodium sulfate. The desiccant was removed, the solvent was evaporated under reduced pressure, and the obtained residue was purified by silica gel column chromatography (hexane-EtOAc) to obtain 83.3 mg of {trans-3-[(tert-butoxycarbonyl)amino]cyclobutyl}methyl methanesulfonate.
Under ice-cooling, to a solution of tert-butyl[2-(4-aminophenyl)ethyl]carbamate (1.5 g) in chloroform (15 ml) were sequentially added TEA (0.973 ml) and methanesulfonyl chloride (0.540 ml), followed by stirring at room temperature for 3 hours. TEA (1.326 ml) and mesyl chloride (0.737 ml) were sequentially added thereto, followed by stirring at room temperature for additional 3 hours. The mixed reaction liquid was concentrated under reduced pressure, and the obtained residue was purified by amino silica gel flash column chromatography (hexane-EtOAc) to obtain 1.03 g of tert-butyl (2-{4-[(methylsulfonyl)amino]phenyl}ethyl)carbamate.
Under ice-cooling, to a solution of tert-butyl piperidin-4-ylcarbamate (400 mg) and DIPEA (0.30 ml) in dichloromethane (6 ml) was added chloroacetyl chloride (0.175 ml), followed by stirring at the same temperature for 30 minutes and at room temperature for 2.5 hours. To the reaction mixture were added EtOAc and 0.5 M hydrochloric acid, followed by liquid separation, and then the organic layer was sequentially washed with water, saturated aqueous sodium bicarbonate, water, and saturated brine, and dried over anhydrous sodium sulfate. After the desiccant was removed, the solvent was evaporated under reduced pressure and the residue was purified by silica gel flash column chromatography (hexane-EtOAc) to obtain 545 mg of tert-butyl[1-(chloroacetyl)piperidin-4-yl]carbamate.
To a solution of 6-chloronicotinonitrile (400 mg) and tert-butylpiperidin-4-ylcarbamate (693 mg) in DMF (4.8 ml) was added potassium carbonate (598 mg), followed by stirring at 120° C. for 2 hours. To the reaction mixture were added EtOAc and water, followed by liquid separation, and then the organic layer was sequentially washed with water (three times) and saturated brine, and dried over anhydrous sodium sulfate. After the desiccant was removed, the solvent was evaporated under reduced pressure, and the precipitated powder was collected by filtration. After washing with EtOAc, the powder was dried to obtain 504 mg of tert-butyl[1-(5-cyanopyridin-2-yl)piperidin-4-yl]carbamate.
Under ice-cooling, to a solution of 4-[({(1S,3R,4S,5S)-4-[(3-{[tert-butyldimethyl)silyl]oxypropyl)amino]adamantan-1-yl}methyl)amino]-2-{[2-(methylsulfanyl)benzyl]amino}pyrimidine-5-carbonitrile (65 mg) in MeOH (1.3 mL) were added a 35% aqueous formalin solution (34 mg) and sodium cyanoborohydride (27 mg), followed by stirring at room temperature for 5 hours. To the reaction mixture was added saturated aqueous sodium bicarbonate (5 mL), and the precipitated white solid was collected by filtration. The obtained solid was dissolved in chloroform and purified by amino silica gel flash column chromatography (hexane-EtOAc) to obtain 60 mg of 4-[({(1S,3R,4S,5S)-4-[(3-[tert-butyldimethyl)silyl]oxypropyl)(methyl)amino]adamantan-1-yl}methyl)amino]-2-{[2-(methylsulfanyl)benzyl]amino}pyrimidine-5-carbonitrile.
To a solution of rel-4-({[(1S,3R,4S,5S)-4-aminoadamantan-1-yl]methyl}amino)-2-{[2-(trifluoromethoxy)benzyl]amino}pyrimidine-5-carbonitrile (100 mg) in EtOAc (1.0 ml) were added tert-butyl (4-oxocyclohexyl)carbamate (67.7 mg) and titanium (IV) isopropoxide (250 μl), followed by stirring at room temperature for 20 minutes. Then, to the mixed reaction liquid was added platinum oxide (12 mg), followed by stirring at room temperature for 220 minutes under a hydrogen atmosphere. To the mixed reaction liquid were sequentially added water and EtOAc, followed by stirring, and the insoluble materials were removed by filtration. The filtrate was concentrated under reduced pressure and the residue was extracted with EtOAc. The organic layer was washed with saturated brine and dried over anhydrous sodium sulfate. After the desiccant was removed, the solvent was evaporated under reduced pressure, and the residue was purified by amino silica gel flash column chromatography (hexane-EtOAc) to first elute tert-butyl rel-(cis-4-{[(1R,2S,3S,5S)-5-{[(5-cyano-2-{[2-(trifluoromethoxy)benzyl]amino}pyrimidin-4-yl)amino]methyl}adamantan-2-yl]amino}cyclohexyl)carbamate (86.6 mg), and then elute tert-butyl rel-(trans-4-{[(1R,2S,3S,5S)-5-{[(5-cyano-2-{[2-(trifluoromethoxy)benzyl]amino}pyrimidin-4-yl)amino]methyl}adamantan-2-yl]amino}cyclohexyl)carbamate (28.4 mg).
Under ice-cooling, to a solution of (4-methoxypyridin-3-yl)methanol (58 mg) in chloroform (0.6 ml) was added thionyl chloride (0.046 ml), followed by stirring at room temperature for 1 hour. The reaction liquid was concentrated under reduced pressure, and to the reside was added saturated aqueous sodium bicarbonate, followed by extraction with EtOAc. The organic layer was washed with water and saturated brine, and dried over anhydrous magnesium sulfate. After the desiccant was removed, the solvent was evaporated under reduced pressure to obtain 65 mg of 3-(chloromethyl)-4-methoxypyridine.
Under ice-cooling, to a solution of tert-butyl 5-cyano-4,5,6,7-tetrahydro-1H-benzimidazole-1-carboxylate (100 mg) in MeOH (3 mL) were slowly added cobalt (II) chloride hexahydrate (192 mg) and sodium borohydride (61 mg), followed by stirring at room temperature for 1 hour. To the reaction liquid was added 1 M hydrochloric acid (1 mL), and the insoluble materials were removed by filtration. The filtrate was washed with chloroform (10 mL), and to the aqueous layer was added 1 M hydrochloric acid (4 mL), followed by concentration under reduced pressure, to obtain 72 mg of 1-(4,5,6,7-tetrahydro-1H-benzimidazol-5-yl)methylamine dihydrochloride.
To a solution of tert-butyl (2-vinylbenzyl)carbamate (30 mg) in water (0.075 ml)-acetone (0.15 ml) were added a 4% aqueous osmium tetroxide solution (41 mg) and 4-methylmorpholine N-oxide (23 mg) at room temperature. After stirring at the same temperature for 2 hours, a 10% aqueous sodium sulfite solution was added thereto under ice-cooling. The mixture was extracted with EtOAc, and the organic layer was washed with saturated brine and dried over anhydrous magnesium sulfate. After the desiccant was removed, the solvent was evaporated under reduced pressure, and the obtained residue was purified by silica gel flash column chromatography (hexane-EtOAc) to obtain 24 mg of tert-butyl[2-(1,2-dihydroxyethyl)benzyl]carbamate.
Under ice-cooling, to a solution of tert-butyl 5-carbamoyl-4,5,6,7-tetrahydro-1H-benzimidazole-1-carboxylate (500 mg) in THF (5 mL) were added trifluoroacetic anhydride (0.32 mL) and pyridine (0.32 mL), followed by stirring at room temperature for 1 hour. To the reaction liquid was added saturated aqueous sodium bicarbonate (10 mL), followed by extraction with EtOAc (40 mL). The organic layer was washed with water (10 mL) and saturated brine (10 mL), and then dried over anhydrous sodium sulfate. Then, the desiccant was removed and the solvent was evaporated under reduced pressure. The obtained residue was purified by silica gel flash column chromatography (hexane-EtOAc) to obtain 380 mg of tert-butyl 5-cyano-4,5,6,7-tetrahydro-1H-benzimidazole-1-carboxylate.
Under ice-cooling, to a solution of 2-(methylsulfanyl)nicotinonitrile (382 mg) in MeOH (6 ml) was added cobalt (II) chloride hexahydrate (1.69 g), and sodium borohydride (346 mg) was added thereto in small portions at the same temperature. After stirring at room temperature for 3 hours, the precipitate was removed by filtration through Celite. The filtrate was concentrated under reduced pressure, and to the residue was added 1 M hydrochloric acid, followed by washing with chloroform. The aqueous layer was alkalified by the addition of 28% aqueous ammonia and then extracted with chloroform, and the organic layer was dried over anhydrous magnesium sulfate. The solvent was evaporated under reduced pressure to obtain 237 mg of 1-[2-(methylsulfanyl)pyridin-3-yl]methylamine.
Under ice-cooling, to a mixed solution of rel-4-({[(1S,3R,4S,5S)-4-(3-hydroxyazetidin-1-yl)adamantan-1-yl]methyl}amino)-2-{[2-(trifluoromethoxy)benzyl]amino}pyrimidine-5-carbonitrile (50 mg) in THF (2 ml) and toluene (3 ml) were added succinimide (19.8 mg), triphenylphosphine (29.8 mg), and DEAD (17.8 μl), followed by stirring at room temperature. Succinimide (33.4 mg), triphenylphosphine (74.4 mg), DEAD (44.5 μl) THF (2 ml), and toluene (1 ml) were added thereto, followed by further stirring at room temperature. After completion of the reaction, the mixed reaction liquid was diluted with EtOAc, and the organic layer was sequentially washed with water (three times) and saturated brine, and dried over anhydrous sodium sulfate. After the desiccant was removed, the solvent was evaporated under reduced pressure, and the obtained residue was purified by silica gel flash column chromatography (hexane-EtOAc) to obtain 70.7 mg of rel-4-[({(1S,3R,4S,5S)-4-[3-(1,3-dioxo-1,3-dihydro-2H-isoindol-2-yl)azetidin-1-yl]adamantan-1-yl}methyl)amino]-2-{[2-(trifluoromethoxy)benzyl]amino}pyrimidine-5-carbonitrile.
Under ice-cooling, to a suspension of rel-(1S,3R,4S,5S)-4-aminoadamantane-1-carboxylic acid (100 mg) in 1,4-dioxane (0.7 ml) was added a 1 M aqueous sodium hydroxide solution (0.62 ml), followed by stirring at the same temperature for 10 minutes for dissolution. Under ice-cooling, a solution of di-tert-butyl dicarbonate (115 mg) in 1,4-dioxane (0.1 ml) was added dropwise thereto, followed by stirring at room temperature for 4 hours. Under ice-cooling, 1 M hydrochloric acid (0.74 ml) was added thereto, followed by extraction with EtOAc, and washing with water (twice) and then with saturated brine. The organic layer was dried over anhydrous sodium sulfate, filtered, and then concentrated under reduced pressure to precipitate crystals. Thus, the crystals were suspended in hexane (3 ml) before dryness, and collected by filtration to obtain 111.1 mg of rel-(1S,3R,4S,5S)-4-[(tert-butoxycarbonyl)amino]adamantane-1-carboxylic acid.
To a suspension of (methoxymethyl)(triphenyl)phosphonium chloride (164.57 g) which had been cooled in a dry ice-acetone bath in THF (500 ml) was added dropwise a solution of n-butyllithium in hexane (concentration 1.65 M, 281.3 ml) at −55° C. or lower under a nitrogen air flow. Then, the mixed reaction liquid was warmed, followed by stirring at room temperature for 1 hour. After stirring, the mixed reaction liquid was cooled under ice, and a solution of 4-hydroxy-4-methylcyclohexanone (20.51 g) in THF (205 ml) was added dropwise thereto. After dropwise addition, the mixed reaction liquid was warmed to room temperature, followed by stirring for 15 hours. To the mixed reaction liquid were sequentially added water and EtOAc, followed by stirring, and then the organic layer was collected by separation. The aqueous layer was further extracted with EtOAc, and the organic layer was combined, washed with saturated brine, and dried over anhydrous sodium sulfate. After the desiccant was removed, the solvent was evaporated under reduced pressure. The residue was purified by silica gel flash column chromatography (hexane-EtOAc) to obtain 4-(methoxymethylene)-1-methylcyclohexanol (21.37 g).
To a solution of tert-butyl rel-[(1R,2S,3S,5S)-5-(azidomethyl)adamantan-2-yl]carbamate (300 mg) in THF (3 ml) was added triphenylphosphine (300 mg), followed by stirring at room temperature for 4 hours. To the reaction mixture was added water (1.8 ml), followed by stirring at room temperature for 2 hours, and then the solvent was evaporated under reduced pressure. The residue was purified by silica gel flash column chromatography (the side products were first eluted with EtOAc alone, and then the eluting solvent was changed to MeOH/chloroform/28% NH3 aq. (1/9/0.1)) to obtain 270 mg of tert-butyl rel-[(1R,2S,3S,5S)-5-(aminomethyl)adamantan-2-yl]carbamate.
To a solution of tert-butyl rel-[(1R,2S,3S,5S)-5-{[(2-chloro-5-fluoropyrimidin-4-yl)amino]methyl}adamantan-2-yl]carbamate (100 mg) and 2-(trifluoromethoxy)benzylamine (280 mg) in DMI (0.8 ml) was added DIPEA (0.127 ml), followed by irradiation with microwaves at 165° C. for 4 hours. The reaction liquid was diluted with EtOAc, sequentially washed with water, saturated aqueous ammonium chloride solution, water, saturated aqueous sodium hydrogen carbonate solution, water, and saturated brine, and dried over anhydrous sodium sulfate. After the desiccant was removed, the solvent was evaporated under reduced pressure. The obtained residue was purified by silica gel flash column chromatography (hexane-EtOAc) to obtain 78.3 mg of tert-butyl rel-[(1R,2S,3S,5S)-5-{[(5-fluoro-2-{[2-(trifluoromethoxy)benzyl]amino}pyrimidin-4-yl)amino]methyl}adamantan-2-yl]carbamate.
Under ice-cooling, to a solution of tert-butyl[(2-chloropyridin-3-yl)methyl]carbamate (450 mg) in DMF (2 ml) were added cyclopentanethiol (0.65 ml) and sodium hydride (about 40% of mineral oil added, 220 mg), followed by stirring at room temperature for 4 hours. Then, the reaction liquid was cooled under ice, and cyclopentanethiol (0.40 ml) and sodium hydride (about 40% of mineral oil added, 140 mg) were added thereto, followed by stirring at room temperature for 2 hours. Again, the reaction liquid was cooled under ice, and saturated aqueous ammonium chloride solution was added thereto, followed by extraction with chloroform. The organic layer was dried over anhydrous sodium sulfate, the desiccant was removed, and then the solvent was evaporated under reduced pressure. The obtained residue was purified by silica gel flash column chromatography (hexane-EtOAc) to obtain 400 mg of tert-butyl{[2-(cyclopentylsulfonyl)pyridin-3-yl]methyl}carbamate.
At room temperature, to a suspension of rel-(1S,3R,4S,5S)-4-[(tert-butoxycarbonyl)amino]adamantane-1-carboxylic acid (99.0 mg) in DME (0.99 ml) was added N-methylmorpholine (0.044 ml) for dissolution. Under ice-cooling, isobutyl chlorocarbonate (0.052 ml) was added dropwise thereto, followed by stirring at the same temperature for 40 minutes. The precipitated white insoluble materials were removed by filtration and washed with DME (0.5 ml). The filtrate was cooled under ice, sodium borohydride (25.3 mg) was added thereto, and then MeOH (0.495 ml) was slowly added dropwise thereto. After stirring at room temperature for 1 hour, the reaction liquid was cooled under ice and diluted with EtOAc. The reaction liquid was acidified by the addition of 1 M hydrochloric acid (1.0 ml), and the organic layer was collected by separation. The organic layer was sequentially washed with water (twice), saturated aqueous sodium bicarbonate, water, and then saturated brine, dried over anhydrous sodium sulfate, filtered, and then concentrated under reduced pressure. The residue was purified by silica gel column chromatography (hexane-EtOAc). A fraction including a desired product was concentrated under reduced pressure, and then the residue was dissolved in EtOAc, and concentrated under reduced pressure to about 0.1 ml. Hexane (1.5 ml) was added portionwise thereto for crystallization, and the crystals were collected by filtration to obtain 77.3 mg of tert-butyl rel-[(1R,2S,3S,5S)-5-(hydroxymethyl)adamantan-2-yl]carbamate.
Rel-(1R,3S,5R,7S)-4-oxoadamantane-1-carboxylic acid (1.0 g) was dissolved in a solution (concentration 8 M, 20 ml) of ammonia in MeOH, and 10% Pd/C (wetted with 50% water, 100 mg) was added thereto, followed by stirring at 25° C. for 10 hours under a hydrogen atmosphere of 3 atm. The product that had been precipitated in a large amount was dissolved in water (20 ml), and the catalyst was removed by filtration through Celite. MeOH was evaporated under reduced pressure, and to the residue was added dropwise acetonitrile (30 ml), followed by stirring at room temperature for 1 hour. The precipitate was collected by filtration, washed with MeCN (10 ml), and then dried under reduced pressure at 45° C. to obtain 982 mg of rel-(1S,3R,5S)-4-aminoadamantane-1-carboxylic acid as a mixture of trans isomer and cis isomer at a ratio of 3.5:1.
Rel-(1S,3R,5S)-4-aminoadamantane-1-carboxylic acid (mixture of trans product and cis product at a ratio of 3.5:1, 100 mg) was suspended in water (4 ml), followed by stirring at 75° C. for 30 minutes. While stirring, the suspension was cooled back to room temperature, and MeCN (4 ml) was slowly added dropwise thereto, followed by stirring at the same temperature for 30 minutes. The precipitate was collected by filtration, washed with acetonitrile (1 ml), and then dried under reduced pressure at 45° C. to obtain 50.0 mg of rel-(1S,3R,4S,5S)-4-aminoadamantane-1-carboxylic acid.
To a solution of 4-(methoxymethylene)-1-methylcyclohexanol (5.0 g) in MeCN (50 ml) were sequentially added water (8.6 ml) and TFA (3.6 ml), followed by stirring at room temperature for 4 hours. The mixed reaction liquid was adjusted to be neutral with saturated aqueous sodium hydrogen carbonate solution, and then extracted with EtOAc four times. The organic layer was combined, washed with saturated brine, and dried over anhydrous magnesium sulfate. After the desiccant was removed, the solvent was evaporated under reduced pressure. The obtained residue was purified by silica gel flash column chromatography (hexane-EtOAc) to first elute cis-4-hydroxy-4-methylcyclohexanecarbaldehyde (2.37 g) and then elute trans-4-hydroxy-4-methylcyclohexanecarbaldehyde (2.7 g).
Each of the Preparation Example compounds was prepared in the same manner as the methods of Preparation Examples above, using each of the corresponding starting materials. The structures, the production processes, and the physicochemical data of the compounds of Preparation Examples are shown in Tables below.
To a solution of benzyl rel-1-[(1R,3S,5S)-5-{[(5-cyano-2-{[2-(trifluoromethoxy)benzyl]amino}pyrimidin-4-yl)amino]methyl}adamantan-2-yl]-D-prolinate (100 mg) in EtOH (5 ml) was added 10% Pd/C (wetted with 50% water, 20 mg), followed by stirring at room temperature at a normal pressure under a hydrogen atmosphere. After completion of the reaction, the catalyst was separated by filtration through Celite, and the filtrate was concentrated under reduced pressure. The obtained residue was purified by preparative alumina thin layer chromatography (chloroform-MeOH) to obtain 74.4 mg of rel-1-[(1R,3S,5S)-5-{[(5-cyano-2-{[2-(trifluoromethoxy)benzyl]amino}pyrimidin-4-yl)amino]methyl}adamantan-2-yl]-D-proline.
To a solution of rel-4-({[(1S,3R,4S,5S)-4-aminoadamantan-1-yl]methyl}amino)-2-{[2-(trifluoromethoxy)benzyl]amino}pyrimidine-5-carbonitrile (50 mg) in DMF (0.2 ml) were added iodobenzene (14.2 lμ), cesium acetate (50.8 mg) and copper (I) iodide (20.2 mg), followed by stirring at 90° C. for 24 hours under irradiation with microwaves. The mixed reaction liquid was diluted with EtOAc, and then the organic layer was sequentially washed with water and saturated brine, and dried over anhydrous sodium sulfate. After the desiccant was removed, the solvent was evaporated under reduced pressure. The obtained residue was purified by preparative silica gel thin layer chromatography (chloroform-MeOH) to obtain 5.1 mg of rel-4-({[(1S,3R,4S,5S)-4-anilinoadamantan-1-yl]methyl}amino)-2-{[2-(trifluoromethoxy)benzyl]amino}pyrimidine-5-carbonitrile.
Under ice-cooling, to a solution of rel-4-({[(1S,3R,4S,5S)-4-aminoadamantan-1-yl]methyl}amino)-2-{[2-(trifluoromethoxy)benzyl]amino}pyrimidine-5-carbonitrile (50 mg) in DMF (1.0 ml) were sequentially added DIPEA (27.7 μl) and acetyl chloride (10.3 μl), followed by stirring at the same temperature for 1 hour. The reaction mixture was diluted with EtOAc, and then the organic layer was sequentially washed with water and saturated brine, and dried over anhydrous sodium sulfate. After the desiccant was removed, the solvent was evaporated under reduced pressure. The obtained residue was purified by preparative silica gel thin layer chromatography (chloroform-MeOH) to obtain 46 mg of rel-N-[(1R,2S,3S,5S)-5-{[(5-cyano-2-{[2-(trifluoromethoxy)benzyl]amino}pyrimidin-4-yl)amino]methyl}adamantan-2-yl]acetamide.
To a solution of rel-(4R)-1-[(1R,3S,5S)-5-{[(5-cyano-2-{[2-(trifluoromethoxy)benzyl]amino}pyrimidin-4-yl)amino]methyl}adamantan-2-yl]-4-hydroxy-D-proline (23 mg) in DMF (0.5 ml) were sequentially added ammonium chloride (6.3 mg), HOBt (15.9 mg), and WSC (18.3 mg), followed by stirring at room temperature overnight. The reaction mixture was diluted with EtOAc, then sequentially washed with water and saturated brine, and dried over anhydrous sodium sulfate. After the desiccant was removed, the solvent was evaporated under reduced pressure. The obtained residue was purified by preparative silica gel thin layer chromatography (THF), and then purified by amino silica gel flash column chromatography (chloroform-MeOH) to obtain 10 mg of rel-(4R)-1-[(1R,3S,5S)-5-{[(5-cyano-2-{[2-(trifluoromethoxy)benzyl]amino}pyrimidin-4-yl)amino]methyl}adamantan-2-yl]-4-hydroxy-D-prolinamide.
To a solution of rel-4-({[(1S,3R,4S,5S)-4-aminoadamantan-1-yl]methyl}amino)-2-{[2-(trifluoromethoxy)benzyl]amino}pyrimidine-5-carbonitrile (50 mg) in DMF (1.0 ml) were sequentially added nicotinic acid (14.3 mg), HOBt (15.7 mg), and WSC (18.1 mg), followed by stirring at room temperature overnight. The reaction mixture was diluted with EtOAc, and then the organic layer was sequentially washed with water and saturated brine, and dried over anhydrous sodium sulfate. After the desiccant was removed, the solvent was evaporated under reduced pressure. The obtained residue was purified by amino silica gel flash column chromatography (hexane-EtOAc) to obtain 58 mg of rel-N-[(1R,2S,3S,5S)-5-{[(5-cyano-2-{[2-(trifluoromethoxy)benzyl]amino}pyrimidin-4-yl)amino]methyl}adamantan-2-yl]nicotinamide.
To a solution of rel-4-({[(1S,3R,4S,5S)-4-aminoadamantan-1-yl]methyl}amino)-2-{[2-(trifluoromethoxy)benzyl]amino}pyrimidine-5-carbonitrile (50 mg) in N,N-dimethylacetamide (2.5 ml) were added sodium carbonate (44.8 mg) and 1,3-dibromopropane (43.2 μl), followed by stirring at 100° C. for 30 minutes under irradiation with microwaves. The reaction mixture was diluted with EtOAc, and then the organic layer was sequentially washed with water and saturated brine, and dried over anhydrous sodium sulfate. After the desiccant was removed, the solvent was evaporated under reduced pressure. The obtained residue was purified by preparative silica gel thin layer chromatography (chloroform-MeOH) to obtain 45 mg of rel-4-({[(1S,3R,4S,5S)-4-azetidin-1-yladamantan-1-yl]methyl}amino)-2-{[2-(trifluoromethoxy)benzyl]amino}pyrimidine-5-carbonitrile.
To a solution of rel-4-({[(1S,3R,4S,5S)-4-aminoadamantan-1-yl]methyl}amino)-2-{[2-(trifluoromethoxy)benzyl]amino}pyrimidine-5-carbonitrile (30 mg) in DMF (0.6 ml) were added DIPEA (22.1 μl) and 2-bromoethanol (4.5 μl), followed by heating and stirring at 60° C. The reaction mixture was diluted with EtOAc, and then the organic layer was sequentially washed with water and saturated brine, and dried over anhydrous sodium sulfate. After the desiccant was removed, the solvent was evaporated under reduced pressure. The obtained residue was purified by preparative silica gel thin layer chromatography (chloroform-MeOH), and then by preparative alumina thin layer chromatography (chloroform-MeOH) to obtain 25 mg of rel-4-[({(1S,3R,4S,5S)-4-[(2-hydroxyethyl)amino]adamantan-1-yl}methyl)amino]-2-{[2-(trifluoromethoxy)benzyl]amino}pyrimidine-5-carbonitrile.
To a solution of 4-({[(1S,3R,4S,5S)-4-aminoadamantan-1-yl]methyl}amino)-2-{[2-(trifluoromethoxy)benzyl]amino}pyrimidine-5-carbonitrile (50 mg) in DMF (1 ml) were added (2,2-dimethyl-1,3-dioxan-5-yl)methyl 4-methylbenzenesulfonate (47.7 mg) and potassium carbonate (29.2 mg), followed by stirring at 70° C. for 12 hours. To the reaction mixture was added water, followed by extraction with EtOAc. The organic layer was dried over magnesium sulfate, the desiccant was removed, and then the solvent was evaporated under reduced pressure. The obtained residue was purified by silica gel column chromatography (hexane-EtOAc) to obtain 21 mg of 4-({[(1S,3R,4S,5S)-4-{[(2,2-dimethyl)-1,3-dioxan-5-yl)methyl]amino}adamantan-1-yl]methyl}amino)-2-{[2-(trifluoromethoxy)benzyl]amino}pyrimidine-5-carbonitrile.
To a mixed solution of rel-4-({[(1S,3R,4S,5S)-4-aminoadamantan-1-yl]methyl}amino)-2-{[2-(trifluoromethoxy)benzyl]amino}pyrimidine-5-carbonitrile (50 mg) in EtOH (1.0 ml) and THF (0.5 ml) was added oxiran-2-ylmethanol (8.2 μl), followed by stirring at room temperature for 24 hours. Oxylan-2-yl methanol (82.1 μl) was added thereto, followed by stirring at room temperature for additional 48 hours. The mixed reaction liquid was purified by amino silica gel flash column chromatography (chloroform-MeOH) to obtain 25 mg of rel-4-[({(1S,3R,4S,5S)-4-[(2,3-dihydroxypropyl)amino]adamantan-1-yl}methyl)amino]-2-{[2-(trifluoromethoxy)benzyl]amino}pyrimidine-5-carbonitrile.
To a solution of rel-4-({[(1S,3R,4S,5S)-4-aminoadamantan-1-yl]methyl}amino)-2-{[2-(trifluoromethoxy)benzyl]amino}pyrimidine-5-carbonitrile (50 mg) in 1,3-dimethylimidazolidin-2-one (1.0 ml) were added DIPEA (36.9 μl) and 2,2,2-trifluoroethyl trifluoromethanesulfonate (18.3 μl), followed by stirring at room temperature. After completion of the reaction, the mixed reaction liquid itself was purified by silica gel flash column chromatography (chloroform-MeOH) to obtain 55 mg of rel-4-[({(1S,3R,4S,5S)-4-[(2,2,2-trifluoroethyl)amino]adamantan-1-yl}methyl)amino]-2-{[2-(trifluoromethoxy)benzyl]amino}pyrimidine-5-carbonitrile.
To a solution of 4-({[(1S,3R,4S,5S)-4-aminoadamantan-1-yl]methyl}amino)-2-{[2-(trifluoromethoxy)benzyl]amino}pyrimidine-5-carbonitrile (50 mg) in 1,3-dimethylimidazolidin-2-one (0.5 ml) were added 2,2-bis(bromomethyl)propane-1,3-diol (221.8 mg), and potassium carbonate (146.3 mg), followed by stirring at room temperature for 4 days. The reaction mixture was purified by silica gel column chromatography (chloroform-MeOH) as it was to obtain 5 mg of [({(1S,3R,4S,5S)-4-[3,3-bis(hydroxymethyl)azetidin-1-yladamantan-1-yl}methyl)amino]-2-{[2-(trifluoromethoxy)benzyl]amino}pyrimidine-5-carbonitrile.
A mixture of steric isomers (39 mg) of rel-4-[({(1R,3S,5R)-4-[(trans-4-hydroxycyclohexyl)amino]adamantan-1-yl}methyl)amino]-2-{[2-(trifluoromethoxy)benzyl]amino}pyrimidine-5-carbonitrile was subjected to separation and purification by reverse phase liquid chromatography (eluent: a mixed liquid of 0.2% formic acid/MeOH and water) to obtain a formate of rel-4-[({(1R,3S,4R,5R)-4-[(trans-4-hydroxycyclohexyl)amino]adamantan-1-yl}methyl)amino]-2-{[2-(trifluoromethoxy)benzyl]amino}pyrimidine-5-carbonitrile and a formate of rel-4-[({(1R,3S,4S,5R)-4-[(trans-4-hydroxycyclohexyl)amino]adamantan-1-yl}methyl)amino]-2-{[2-(trifluoromethoxy)benzyl]amino}pyrimidine-5-carbonitrile as single isomers, respectively. Then, their formates were each purified by amino silica gel flash column chromatography (hexane-EtOAc) to obtain 19.1 mg of rel-4-[({(1R,3S,4R,5R)-4-[(trans-4-hydroxycyclohexyl)amino]adamantan-1-yl}methyl)amino]-2-{[2-(trifluoromethoxy)benzyl]amino}pyrimidine-5-carbonitrile and 10.4 mg of rel-4-[({(1R,3S,4S,5R)-4-[(trans-4-hydroxycyclohexyl)amino]adamantan-1-yl}methyl)amino]-2-{[2-(trifluoromethoxy)benzyl]amino}pyrimidine-5-carbonitrile.
To a solution of rel-trans-4-{[(1R,2S,3S,5S)-5-(aminomethyl)adamantan-2-yl]amino}cyclohexanol (80 mg) in 1,3-dimethylimidazolidin-2-one (1.0 ml) were added DIPEA (0.20 ml) and 4-chloro-2-{[2-(methylsulfanyl)benzyl]amino}pyrimidine-5-carbonitrile (108.6 mg), followed by stirring at room temperature overnight. The mixed reaction liquid was purified by amino silica gel flash column chromatography (chloroform-MeOH) to obtain 73.4 mg of rel-4-[({(1R,3S,4R,5R)-4-[(trans-4-hydroxycyclohexyl)amino]adamantan-1-yl]methyl)amino}-2-{[2-(methylsulfanyl)benzyl]amino}pyrimidine-5-carbonitrile.
To a solution of ethyl rel-N-[(1R,2S,3S,5S)-5-{[(5-cyano-2-{[2-(trifluoromethoxy)benzyl]amino}pyrimidin-4-yl)amino]methyl}adamantan-2-yl]glycinate (24 mg) in THF (2.4 ml) was added a 4 M aqueous lithium hydroxide solution (0.1 ml), followed by stirring at room temperature. After completion of the reaction, to the mixed reaction liquid were added 1 M hydrochloric acid (0.4 ml), followed by stirring, and then the mixed reaction liquid was concentrated under reduced pressure. To the obtained residue was added water, and the solid was collected by filtration and then dried under reduced pressure. The obtained solid was suspended by the addition of EtOAc and diisopropyl ether, and then the solid was collected by filtration and dried under reduced pressure to obtain 11.0 mg of rel-N-[(1R,2S,3S,5S)-5-{[(5-cyano-2-{[2-(trifluoromethoxy)benzyl]amino}pyrimidin-4-yl)amino]methyl}adamantan-2-yl]glycine.
To a solution of rel-4-({[(1S,3R,5S)-4-oxoadamantan-1-yl]methyl}amino)-2-{[2-(trifluoromethoxy)benzyl]amino}pyrimidine-5-carbonitrile (40 mg) in dichloromethane (0.8 ml) were added trans-4-aminocyclohexanol (14.7 mg) and sodium triacetoxyborohydride (53.9 mg), followed by stirring at room temperature overnight. To the mixed reaction liquid was added saturated aqueous sodium bicarbonate, followed by extraction with EtOAc, and then the organic layer was washed with saturated brine, and dried over anhydrous sodium sulfate. After the desiccant was removed, the solvent was evaporated under reduced pressure. The obtained residue was purified by preparative silica gel thin layer chromatography (chloroform-MeOH) to obtain 18 mg of rel-4-[({(1R,3S,5R)-4-[(trans-4-hydroxycyclohexyl)amino]adamantan-1-yl}methyl)amino]-2-{[2-(trifluoromethoxy)benzyl]amino}pyrimidine-5-carbonitrile.
To a solution of rel-4-({[(1S,3R,4S,5S)-4-aminoadamantan-1-yl]methyl}amino)-2-{[2-(methylsulfanyl)benzyl]amino}pyrimidine-5-carbonitrile (50 mg) in dichloromethane (3.5 ml) were added 4-hydroxycyclohexanone (39.4 mg) and sodium triacetoxyborohydride (146.3 mg), followed by stirring at room temperature for 2 hours. To the mixed reaction liquid was added saturated aqueous sodium bicarbonate, followed by extraction with EtOAc, then washing with saturated brine, and drying over anhydrous sodium sulfate. After the desiccant was removed, the solvent was evaporated under reduced pressure. The obtained residue was purified by amino silica gel flash column chromatography (chloroform-MeOH) to obtain 60.6 mg of rel-4-[({(1S,3R,4S,5S)-4-[(4-hydroxycyclohexyl)amino]adamantan-1-yl}methyl)amino]-2-{[2-(methylsulfanyl)benzyl]amino}pyrimidine-5-carbonitrile.
To a solution of rel-4-[({(1S,3R,4S,5S)-4-[(4-hydroxycyclohexyl)amino]adamantan-1-yl}methyl)amino]-2-{[2-(trifluoromethoxy)benzyl]amino}pyrimidine-5-carbonitrile (50 mg) in MeOH (2 ml) were added a 37% aqueous formalin solution (28.4 μl) and sodium cyanoborohydride (8.3 mg), followed by stirring at room temperature for 5 hours. To the mixed reaction liquid was added saturated aqueous sodium bicarbonate, followed by extraction with EtOAc, and then washed with saturated brine, and dried over anhydrous sodium sulfate. After the desiccant was removed, the solvent was evaporated under reduced pressure. The obtained residue was purified by amino silica gel flash column chromatography (chloroform-MeOH) to obtain 37.6 mg of rel-4-[({(1S,3R,4S,5S)-4-[(4-hydroxycyclohexyl)(methyl)amino]adamantan-1-yl}methyl)amino]-2-{[2-(trifluoromethoxy)benzyl]amino}pyrimidine-5-carbonitrile.
To a solution of rel-4-({[(1S,3R,5S)-4-oxoadamantan-1-yl]methyl}amino)-2-{[2-(trifluoromethoxy)benzyl]amino}pyrimidine-5-carbonitrile (30 mg) in MeOH (3 ml) was added sodium borohydride (13.5 mg), followed by stirring at room temperature. After completion of the reaction, the reaction mixture was diluted with EtOAc, and then the organic layer was sequentially washed with water and saturated brine, and dried over anhydrous sodium sulfate. After the desiccant was removed, the solvent was evaporated under reduced pressure. The obtained residue was purified by preparative silica gel thin layer chromatography (chloroform-MeOH) to obtain 25.6 mg of rel-4-({[(1S,3R,5S)-4-hydroxyadamantan-1-yl]methyl}amino)-2-{[2-(trifluoromethoxy)benzyl]amino}pyrimidine-5-carbonitrile.
Under ice-cooling, to a solution of rel-2-[(2-methoxybenzyl)amino]-4-[({(1S,3R,4S,5S)-4-[(4-oxocyclohexyl)amino]adamantan-1-yl}methyl)amino]pyrimidine-5-carbonitrile (40 mg) in THF (2 ml) was added dropwise a 0.97 M solution (0.40 ml) of methylmagnesium bromide in THF under a nitrogen atmosphere, followed by stirring at the same temperature for 3 hours. To the mixed reaction liquid was added water, followed by extraction with EtOAc, and then the organic layer was washed with saturated brine, and dried over anhydrous sodium sulfate. After the desiccant was removed, the solvent was evaporated under reduced pressure. The obtained residue was purified by preparative silica gel thin layer chromatography (chloroform-MeOH-28% aqueous ammonia) to first obtain 4.3 mg of rel-4-[({(1S,3R,4S,5S)-4-[(4-hydroxy-4-methylcyclohexyl)amino]adamantan-1-yl}methyl)amino]-2-[(2-methoxybenzyl)amino]pyrimidine-5-carbonitrile (isomer A) and then obtain 5.3 mg of rel-4-[({(1S,3R,4S,5S)-4-[(4-hydroxy-4-methylcyclohexyl)amino]adamantan-1-yl}methyl)amino]-2-[(2-methoxybenzyl)amino]pyrimidine-5-carbonitrile (isomer B).
To a solution of 4-[(1-azabicyclo[2.2.2]oct-3-ylmethyl)amino]-2-{[2-(trifluoromethoxy)benzyl]amino}pyrimidine-5-carbonitrile (40 mg) in dichloromethane (1 ml) was added 77% MCPBA (contains water, 21 mg) under ice-cooling, followed by stirring at room temperature for 3 hours. To the reaction mixture was added saturated aqueous sodium hydrogen carbonate solution, followed by extraction with EtOAc. The organic layer was dried over anhydrous sodium sulfate, the desiccant was removed, and then the solvent was evaporated under reduced pressure. The obtained residue was purified by amino silica gel flash column chromatography (chloroform-MeOH) to obtain 19 mg of 4-[(1-oxide-azabicyclo[2.2.2]oct-3-ylmethyl)amino]-2-{[2-(trifluoromethoxy)benzyl]amino}pyrimidine-5-carbonitrile.
To a solution of rel-4-({[(1S,3R,4S,5S)-4-(tetrahydro-2H-thiopyran-4-ylamino)adamantan-1-yl]methyl}amino)-2-{[2-(trifluoromethoxy)benzyl]amino}pyrimidine-5-carbonitrile (88 mg) in dichloromethane (2 ml) was added 75% MCPBA (contains water, 106.1 mg), followed by stirring at room temperature for 6 hours. To the mixed reaction liquid was added saturated aqueous sodium bicarbonate, followed by extraction with EtOAc, and the organic layer was washed with saturated brine and then dried over anhydrous sodium sulfate. After the desiccant was removed, the solvent was evaporated under reduced pressure. The obtained residue was purified by silica gel flash column chromatography (chloroform-MeOH), and then by preparative silica gel thin layer chromatography (chloroform-MeOH) to obtain 8.8 mg of rel-4-[({(1S,3R,4S,5S)-4-[(1,1-dioxidotetrahydro-2H-thiopyran-4-yl)amino]adamantan-1-yl}methyl)amino]-2-{[2-(trifluoromethoxy)benzyl]amino}pyrimidine-5-carbonitrile.
To a solution of 4-{[(3-endo)-8-benzyl-8-azabicyclo[3.2.1]oct-3-yl]amino}-2-{[2-(trifluoromethoxy)benzyl]amino}pyrimidine-5-carbonitrile (16 mg) in MeOH (1 ml) were added ammonium formate (100 mg) and a catalytic amount of 10% Pd/C (wetted with 50% water), followed by heating and refluxing for 6 hours. The mixed reaction liquid was left to be cooled to room temperature, then the catalyst was removed, and the filtrate was concentrated under reduced pressure. The obtained residue was dissolved in EtOAc, sequentially washed with water and saturated brine, and dried over anhydrous sodium sulfate. After the desiccant was removed, the solvent was evaporated under reduced pressure. The obtained residue was purified by preparative silica gel thin layer chromatography (chloroform-MeOH) to obtain 8.0 mg of 4-[(3-endo)-8-azabicyclo[3.2.1]oct-3-ylamino]-2-{[2-(trifluoromethoxy)benzyl]amino}pyrimidine-5-carbonitrile.
To a solution of tert-butyl rel-{(1R,2S,3S,5S)-5-[({2-[(2-chlorobenzyl)amino]-5-cyanopyrimidin-4-yl}amino)methyl]adamantan-2-yl}carbamate (70 mg) in dichloromethane (1 ml) was added trifluoroacetic acid (0.135 ml), followed by stirring at room temperature overnight. The mixed reaction liquid was concentrated under reduced pressure, and to the obtained residue was added an aqueous potassium carbonate solution, followed by stirring at room temperature for 3 hours. The precipitated solid was collected by filtration, washed with water, and then dried under reduced pressure to obtain 55 mg of rel-4-({[(1S,3R,4S,5S)-4-aminoadamantan-1-yl]methyl}amino)-2-[(2-chlorobenzyl)amino]pyrimidine-5-carbonitrile.
To a solution of tert-butyl 3-{[(5-cyano-2-{[2-(trifluoromethoxy)benzyl]amino}pyrimidin-4-yl)amino]methyl}-8-azabicyclo[3.2.1]octane-8-carboxylate in dioxane (0.6 ml) was added a 4 M hydrogen chloride dioxane solution (0.28 ml), followed by stirring at room temperature for 10 hours. The reaction mixture was concentrated under reduced pressure to obtain 56 mg of 4-[(8-azabicyclo[3.2.1]oct-3-ylmethyl)amino]-2-{[2-(trifluoromethoxy)benzyl]amino}pyrimidine-5-carbonitrile dihydrochloride.
To a solution of 4-({[1-(tetrahydro-2H-pyran-2-yl)-1H-benzimidazol-5-yl]methyl}amino)-2-{[2-(trifluoromethoxy)benzyl]amino}pyrimidine-5-carbonitrile (50 mg) in EtOH (3 ml) was added 1 M hydrochloric acid (1 ml), followed by stirring at 60° C. for 24 hours. To the mixed reaction liquid were added 1 M hydrochloric acid (1 ml), followed by stirring at 60° C. for additional 24 hours. The reaction liquid was concentrated under reduced pressure, and to the obtained residue were added EtOAc and EtOH. The precipitated solid was collected by filtration, washed with EtOAc, and then dried under reduced pressure to obtain 53.1 mg of 4-[(1H-benzimidazol-5-ylmethyl)amino]-2-{[2-(trifluoromethoxy)benzyl]amino}pyrimidine-5-carbonitrile dihydrochloride.
To a solution of benzyl rel-[(1R,2S,3S,5S)-5-{[(5-cyano-2-{[2-(trifluoromethoxy)benzyl]amino}pyrimidin-4-yl)amino]methyl}adamantan-2-yl]carbamate (55 mg) in MeOH (3 ml) was added 10% Pd/C (wetted with 50% water, 15 mg), followed by stirring at room temperature for 6 hours at a normal pressure under a hydrogen atmosphere. The catalyst was removed by filtration, and the filtrate was concentrated under reduced pressure. The obtained residue was purified by preparative alumina thin layer chromatography (chloroform-MeOH) to obtain 20.0 mg of rel-4-({[(1S,3R,4S,5S)-4-aminoadamantan-1-yl]methyl}amino)-2-{[2-(trifluoromethoxy)benzyl]amino}pyrimidine-5-carbonitrile.
To a solution of benzyl rel-{(1R,2S,3S,5S)-5-[({5-cyano-2-[(2,5-dichlorobenzyl)amino]pyrimidin-4-yl}amino)methyl]adamantan-2-yl}carbamate (45 mg) in acetic acid (1.5 ml) was added 48% hydrobromic acid (1.5 ml), followed by stirring at room temperature for 24 hours. The reaction liquid was concentrated under reduced pressure, then the residue was dissolved in EtOAc, sequentially washed with an aqueous potassium carbonate solution, water, and saturated brine, and dried over anhydrous sodium sulfate. After the desiccant was removed, the solvent was evaporated under reduced pressure. The obtained residue was purified by amino silica gel flash column chromatography (chloroform-MeOH) to obtain 18 mg of rel-4-({[(1S,3R,4S,5S)-4-aminoadamantan-1-yl]methyl}amino)-2-[(2,5-dichlorobenzyl)amino]pyrimidine-5-carbonitrile.
To a solution of rel-4-({[(1S,3R,4S,5S)-4-{[(2-phenyl-1,3-dioxan-4-yl)methyl]amino}adamantan-1-yl]methyl}amino)-2-{[2-(trifluoromethoxy)benzyl]amino}-5-carbonitrile (54 mg) in THF (1.5 ml) was added 1 M hydrochloric acid (1.5 ml), followed by stirring at room temperature for 4 hours. The reaction mixture was cooled under ice, and saturated aqueous sodium bicarbonate was added thereto. The precipitate was collected by filtration and dried under reduced pressure to obtain a solid, which was purified by amino silica gel column chromatography (chloroform-MeOH) to obtain 41.2 mg of rel-4-[({(1S,3R,4S,5S)-4-[(2,4-dihydroxybutyl)amino]adamantan-1-yl}methyl)amino]-2-{[2-(trifluoromethoxy)benzyl]amino}pyrimidine-5-carbonitrile.
To a solution of rel-4-[({(1S,3R,4S,5S)-4-[(2,2-dimethyl-1,3-dioxan-5-yl)amino]adamantan-1-yl}methyl)amino]-2-{[2-(trifluoromethoxy)benzyl]amino}pyrimidine-5-carbonitrile (55 mg) in THF (1.0 ml) was added 1 M hydrochloric acid (1.0 ml), followed by stirring at room temperature. After completion of the reaction, the mixed reaction liquid was concentrated under reduced pressure, and then to the residue was added dichloromethane, followed by concentration under reduced pressure. To the obtained residue was added diethyl ether, and the precipitated solid was collected by filtration, washed with diethyl ether, and then dried under reduced pressure to obtain 50.5 mg of rel-4-({[(1S,3R,4S,5S)-4-{[2-hydroxy-1-(hydroxymethyl)ethyl]amino}adamantan-1-yl]methyl}amino)-2-{[2-(trifluoromethoxy)benzyl]amino}pyrimidine-5-carbonitrile dihydrochloride.
Under ice-cooling, to a solution of rel-4-({[(1S,3R,4S,5S)-4-aminoadamantan-1-yl]methyl}amino)-2-{[2-(trifluoromethoxy)benzyl]amino}pyrimidine-5-carbonitrile (50 mg) in DMF (1.0 ml) were sequentially added DIPEA (27.7 μl) and methanesulfonyl chloride (8.6 μL), followed by stirring at the same temperature for 1 hour. The mixed reaction liquid was diluted with EtOAc, and the organic layer was sequentially washed with water and saturated brine, and dried over anhydrous sodium sulfate. After the desiccant was removed, the solvent was evaporated under reduced pressure. The obtained residue was purified by preparative silica gel thin layer chromatography (chloroform-MeOH) to obtain 31 mg of rel-N-[(1R,2S,3S,5S)-5-{[(5-cyano-2-{[2-(trifluoromethoxy)benzyl]amino}pyrimidin-4-yl)amino]methyl}adamantan-2-yl]methane sulfonamide.
Under ice-cooling, to a solution of rel-2-[(2-chlorobenzyl)amino]-4-({[(1S,3R,5S)-4-piperazin-1-yladamantan-1-yl]methyl}amino)pyrimidine-5-carbonitrile (18 mg) in DMF (360 μl) were sequentially added triethylamine (3.6 μl) and acetic anhydride (7.6 μl), followed by stirring at room temperature. After completion of the reaction, the mixed reaction liquid was diluted with EtOAc, washed with water (three times) and saturated brine, and dried over anhydrous sodium sulfate. The desiccant was removed, then the solvent was evaporated under reduced pressure, and the residue was purified by amino silica gel flash column chromatography (hexane-EtOAc) to obtain 16.2 mg of rel-4-({[(1S,3R,5S)-4-(4-acetylpiperazin-1-yl)adamantan-1-yl]methyl}amino)-2-[(2-chlorobenzyl)amino]pyrimidine-5-carbonitrile.
To a solution of rel-2-[(2-chlorobenzyl)amino]-4-[({(1S,3R,4S,5S)-4-[(2-chloroethyl)amino]adamantan-1-yl}methyl)amino]pyrimidine-5-carbonitrile (27.0 mg) and (2S)-pyrrolidin-2-ylmethanol (7.9 mg) in DMF (0.27 ml) were added potassium iodide (13.8 mg) and DIPEA (0.02 ml), followed by stirring at 75° C. for 2 hours. Then, (2S)-pyrrolidin-2-ylmethanol (1.1 mg) was added thereto, followed by stirring at the same temperature for additional 2 hours. The solvent was evaporated under reduced pressure and the obtained residue was purified by amino silica gel flash column chromatography (chloroform-MeOH) to obtain 25.4 mg of 2-[(2-chlorobenzyl)amino]-4-({[(1R,3R,4S,5S)-4-({2-[(2S)-2-(hydroxymethylpyrrolidin-1-yl]ethyl}amino)adamantan-1-yl]methyl}amino)pyrimidine-5-carbonitrile.
To a solution of rel-4-[({(1R,3S,4R,5R)-4-[(cis-4-aminocyclohexyl)amino]adamantan-1-yl}methyl)amino]-2-[(2-chlorobenzyl)amino]pyrimidine-5-carbonitrile (30 mg) in DMI (1.0 ml) were added DIPEA (80.4 μl) and 2-bromoethanol (8.2 μl), followed by heating and stirring at 120° C. After completion of the reaction, the reaction mixture was diluted with chloroform and purified by amino silica gel flash column chromatography (chloroform-MeOH) as it was to obtain 17.6 mg of rel-2-[(2-chlorobenzyl)amino]-4-({[(1R,3S,4R,5R)-4-({cis-4-[(2-hydroxyethyl)amino]cyclohexyl}amino)adamantan-1-yl]methyl}amino)pyrimidine-5-carbonitrile.
To a solution of rel-2-[(2-chlorobenzyl)amino]-4-[({(1S,3R,4S,5S)-4-[(4-oxocyclohexyl)amino]adamantan-1-yl}methyl)amino]pyrimidine-5-carbonitrile (50 mg) in dichloromethane (3.0 ml) were sequentially added 4,4-difluoropiperidine hydrochloride (30.4 mg), triethylamine (26.7 μl), and sodium triacetoxyborohydride (61.2 mg), followed by stirring at room temperature. After completion of the reaction, to the mixed reaction liquid was added saturated aqueous sodium bicarbonate, followed by extraction with chloroform, and the solvent was evaporated under reduced pressure. The obtained residue was purified by amino silica gel flash column chromatography (chloroform-MeOH) to obtain 13.1 mg of rel-2-[(2-chlorobenzyl)amino]-4-({[(1S,3R,4S,5S)-4-{[4-(4,4-difluoropiperidin-1-yl)cyclohexyl]amino}adamantan-1-yl]methyl}amino)pyrimidine-5-carbonitrile.
To a solution of rel-4-[({(1R,3S,4R,5R)-4-[(cis-4-aminocyclohexyl)amino]adamantan-1-yl}methyl)amino]-2-[(2-chlorobenzyl)amino]pyrimidine-5-carbonitrile (40 mg) in dichloromethane (2.0 ml) were sequentially added 1-acetylpiperidin-4-one (21.7 mg) and sodium triacetoxyborohydride (48.9 mg), followed by stirring at room temperature. After completion of the reaction, to the mixed reaction liquid was added saturated aqueous sodium bicarbonate, followed by extraction with chloroform, and the solvent was evaporated under reduced pressure. The obtained residue was purified by amino silica gel flash column chromatography (chloroform-MeOH) to obtain 54 mg of rel-4-({[(1R,3S,4R,5R)-4-({cis-4-[(1-acetylpiperidin-4-yl)amino]cyclohexyl}amino)adamantan-1-yl]methyl}amino)-2-[(2-chlorobenzyl)amino]pyrimidine-5-carbonitrile.
To a suspension of rel-4-({[(1S,3R,4S,5S)-4-({[trans-4-({[tert-butyl (dimethyl)silyl]oxy}methyl)cyclohexyl]methyl}amino)adamantan-1-yl]methyl}amino)-2-[(2-cyanobenzyl)amino]pyrimidine-5-carbonitrile (68 mg) in MeOH (1.4 ml) was added 1 M hydrochloric acid (0.6 ml), followed by stirring at room temperature for 1 hour. The solvent was evaporated under reduced pressure, and then the residue was diluted with chloroform, and saturated aqueous sodium bicarbonate was added thereto under ice-cooling. The mixture was extracted with a mixed solvent of chloroform-MeOH (10:1), and the organic layer was washed with water and saturated brine, and dried over anhydrous magnesium sulfate. After the desiccant was removed, the solvent was evaporated under reduced pressure and the obtained residue was purified by amino silica gel flash column chromatography (chloroform-MeOH) to obtain 49.4 mg of rel-2-[(2-cyanobenzyl)amino]-4-({[(1S,3R,4S,5S)-4-({[trans-4-(hydroxymethyl)cyclohexyl]methyl}amino)adamantan-1-yl]methyl}amino)pyrimidine-5-carbonitrile.
Under ice-cooling, to a solution of 4-[({(1S,3R,4S,5S)-4-[(3-{[tert-butyldimethyl)silyl]oxypropyl)(methyl)amino]adamantan-1-yl}methyl)amino]-2-{[2-(methylsulfanyl)benzyl]amino}pyrimidine-5-carbonitrile (55 mg) in THF (2 mL) were added a solution (0.20 mL) of 1 M tetrabutylammonium fluoride in THF, followed by stirring at room temperature for 1 hour. The solvent was evaporated under reduced pressure, and purified by amino silica gel flash column chromatography (chloroform-MeOH) as it was to obtain 25 mg of 4-[({(1S,3R,4S,5S)-4-[(3-hydroxypropyl)(methyl)amino]adamantan-1-yl}methyl)amino]-2-{[2-(methylsulfanyl)benzyl]amino}pyrimidine-5-carbonitrile.
To a mixed solution of rel-4-[({(1S,3R,4S,5S)-4-[3-(1,3-dioxo-1,3-dihydro-2H-isoindol-2-yl)azetidin-1-yl]adamantan-1-yl}methyl)amino]-2-{[2-(trifluoromethoxy)benzyl]amino}pyrimidine-5-carbonitrile (64 mg) in EtOH (1.28 ml) and THF (1.28 ml) was added hydrazine monohydrate (18.9 μl), followed by heating and refluxing, and the insoluble materials were removed by filtration, and then the filtrate was concentrated under reduced pressure. To the obtained residue was added chloroform, followed by washing with water twice and drying over anhydrous sodium sulfate. After the desiccant was removed, the solvent was evaporated under reduced pressure and the residue was purified by amino silica gel flash column chromatography (chloroform-MeOH) to obtain 14 mg of rel-4-({[(1S,3R,4S,5S)-4-(3-aminoazetidin-1-yl)adamantan-1-yl]methyl}amino)-2-{[2-(trifluoromethoxy)benzyl]amino}pyrimidine-5-carbonitrile.
To a solution of rel-2-[(2-chlorobenzyl)amino]-4-[({(1S,3R,4S,5S)-4-[(piperidin-4-ylmethyl)amino]adamantan-1-yl}methyl)amino]pyrimidine-5-carbonitrile (40 mg) in DMI (1 ml) were added 1-fluoro-3-iodopropane (18 mg) and DIPEA (0.017 ml), followed by irradiation with microwaves at 100° C. for 1 hour. The reaction mixture was diluted with chloroform, and purified by amino silica gel flash column chromatography (hexane-EtOAc) as it was to obtain 20 mg of rel-2-[(2-chlorobenzyl)amino]-4-({[(1S,3R,4S,5S)-4-({[1-(3-fluoropropyl)piperidin-4-ylmethyl]methyl}amino)adamantan-1-yl}methyl}amino)pyrimidine-5-carbonitrile.
Under ice-cooling, to a solution of tert-butyl 4-{4-[(5-cyano-2-{[2-(trifluoromethoxy)benzyl]amino}pyrimidin-4-yl)amino]butanolyl}piperazine-1-carboxylate (205 mg) in dichloromethane (4.1 ml) was added TFA (0.75 ml), followed by stirring at room temperature for 18 hours. The mixed reaction liquid was concentrated, and an aqueous potassium carbonate solution was added thereto under ice-cooling, followed by stirring at room temperature. The precipitated solid was collected by filtration, dried, and then purified by amino silica gel flash column chromatography (chloroform-MeOH). A fraction including a desired compound was concentrated, and to the residue was added a 4 M hydrogen chloride ethyl acetate solution, followed by concentration and solidification, to obtain 65.6 mg of 4-{[4-oxo-4-(piperazin-1-yl)butyl]amino}-2-{[2-(trifluoromethoxy)benzyl]amino}pyrimidine-5-carbonitrile dihydrochloride.
To a solution of rel-4-[({(1S,3R,4S,5S)-4-[(1,4-dioxaspiro[4.5]dec-8-ylmethyl)amino]adamantan-1-yl}methyl)amino]-2-({[2-(methylsulfanyl)pyridin-3-yl]methyl}amino)pyrimidine-5-carbonitrile (140 mg) in THF (3 ml) was added 1 M hydrochloric acid (2 ml), followed by stirring at room temperature for 2 hours. The reaction mixture was concentrated under reduced pressure, and then saturated aqueous sodium bicarbonate was added thereto, followed by extraction with EtOAc. The organic layer was washed with saturated brine and dried over anhydrous sodium sulfate. The desiccant was removed, then the solvent was evaporated under reduced pressure, and the residue was purified by amino silica gel flash column chromatography (chloroform-MeOH) to obtain 100 mg of rel-2-({[2-(methylsulfanyl)pyridin-3-yl]methyl}amino)-4-({[(1S,3R,4S,5S)-4-{[(4-oxocyclohexyl)methyl]amino}adamantan-1-yl]methyl}amino)pyrimidine-5-carbonitrile.
Each of the Example compounds was prepared in the same manner as the methods of Examples above, using each of the corresponding starting materials. The structures, the production processes, and the physicochemical data of the Example compounds are shown in Tables below.
The compounds shown in Tables below can be prepared using each of the corresponding starting materials, in the same manner as the methods of Preparation Examples and Examples above.
The compound of the formula (I) or a pharmaceutically acceptable salt has a PKCθ inhibition action and can be used as an inhibitor of acute rejection occurring in transplantation.
| Number | Date | Country | Kind |
|---|---|---|---|
| 2008 223323 | Sep 2008 | JP | national |
| 2009121482 | May 2009 | JP | national |
| Filing Document | Filing Date | Country | Kind | 371c Date |
|---|---|---|---|---|
| PCT/JP2009/065149 | 8/31/2009 | WO | 00 | 2/28/2011 |
