JP2009234949A - Plk-1 inhibitor - Google Patents

Abstract

<P>PROBLEM TO BE SOLVED: To provide a compound having a Polo-like kinase-1 inhibition effect or its pharmacologically acceptable salt. <P>SOLUTION: A heterocyclic compound is represented by the chemical formula (wherein the heterocycle represented by A is pyridine, pyrimidine, dihydropyrimidine, benzimidazole, benzothiazole, dihydrothiazole, or thiazoline; the ring represented by B is benzene or furan; R<SP>1</SP>is a nitro group, an oxo group; a lower alkoxy group, a carbonyl group, a lower alkoxycarbonyl group, a lower alkanoyl group, an amino group, a thioxo group or a cyano group; R<SP>2</SP>is a halogen, a lower alkoxy group, a lower alkoxycarbonyl group, a lower alkanoyl group or a cyano group; X is -S-, -O-, or -NH-; l is an integer of 0-3; m is an integer of 0 or 1; n is an integer of 0-3; and p is an integer of 0-3) and includes its salt. <P>COPYRIGHT: (C)2010,JPO&INPIT

Description

  The present invention relates to a compound that inhibits polo-like kinase-1 (hereinafter sometimes referred to as Plk-1), a pharmaceutically acceptable salt thereof, and a medicament containing them.

Cells proliferate by repeating division, but they do not repeat division randomly, but proliferate by self-diagnosis and control of normal division in the cell cycle. On the other hand, if this control breaks down for some reason, the cells proliferate in a disorderly manner, and it is considered that the disease falls into a disease characterized by abnormal proliferation of cells such as cancer and rheumatoid arthritis.
The M phase (cell division) in the cell cycle is an important period for distributing the chromosomes of cells and transmitting accurate genetic information to daughter cells. Therefore, progression from the G2 phase (the time when cells are ready to enter division) to the M phase is strictly controlled by various cell cycle regulators. Polo-like kinase-1 (Plk-1) is a serine / threonine kinase involved in the control of G2 / M phase. The Cdc2-cyclin B complex plays a key role in the transition from G2 phase to M phase, and cells cannot move from G2 phase to M phase when Cdc2 is phosphorylated . Although Wee1 is involved in this phosphorylation of Cdc2, Plk-1 is inhibited by phosphorylating Wee1. In addition, phosphatase Cdc25 is involved in dephosphorylation of phosphorylated Cdc2, but Plk-1 is activated by phosphorylating Cdc25. When phosphate is removed from Cdc2 due to inhibition of Weel by Plk-1 and activation of Cdc25, the Cdc2-cyclin B complex moves into the nucleus, and progression from the G2 phase to the M phase starts. Plk-1 is directly involved in the nuclear translocation of the Cdc2-cyclin B complex by phosphorylating cyclin B.

  As described above, since Plk-1 is involved in various stages of Cdc2-cyclin B complex activation, which is the key to progression from G2 phase to M phase, inhibitors against Plk-1 are cancer cells. Is thought to stop the growth of G2 in the G2 / M phase. In fact, it has been experimentally proved that siRNA against Plk-1 induces G2 / M phase arrest in cancer cells and induces apoptosis (Non-patent Document 1). In addition, it is reported that Plk-1 is highly expressed in many cancers compared to normal tissues (Non-patent Document 2), and that Plk-1 is highly expressed in metastasized cancers. (Non-patent Document 3), Plk-1 is considered to be an attractive target molecule in cancer treatment, and a compound that inhibits it is expected as a novel anticancer agent.

  In addition, the involvement of Plk-1 extends to areas other than cancer. For example, Plk-1 is important in the development of these diseases, such as synovial cells derived from rheumatoid arthritis patients (Non-patent Document 4) and high expression in the brains of Alzheimer patients (Non-patent Document 5). It is becoming clear that it plays a role. From these information, it is considered that Plk-1 can be a target molecule for the treatment of diseases associated with abnormal cell regulation such as rheumatoid arthritis and Alzheimer as well as the cancer treatment region.

To date, microtubules such as taxanes and vinca alkaloids have already been clinically applied as drugs that act in the G2 / M phase, but serious side effects are problematic. Therefore, there is a need for new treatment methods targeting the G2 / M phase.
Further, Plk-1 inhibitors currently known include scytonemin isolated from cyanobacteria, staurosporine, a broad protein kinase inhibitor, puravalanol A, a cyclin-dependent kinase inhibitor, phosphatidylinositol- LY294002, a 3-kinase inhibitor, and morin, quercetin, etc. of flavonoids have been reported (Non-patent Document 6). Development of Plk-1 inhibitors is desired.
Proc.Natl.Acad.Sci.USA, 2003, 100: 5789-5794 Proc. Natl. Acad. Sci. USA, 1994, 91: 1736-1740 J. Cutan. Pathol., 2002, 29: 354-358 Biochem. Biophys. Res. Commun. 2007, 357: 353-359 Neurobiol. Aging, 2000, 21: 837-841 Curr. Top. Med. Chem., 2005, 5: 181-197

    Accordingly, an object of the present invention is to provide a compound having an excellent Plk-1 inhibitory effect or a pharmaceutically acceptable salt thereof, and a medicament containing the compound.

  In order to solve the above-mentioned problems, the present inventor screened a wide range of compounds using Polo-like kinase 1 Assay / Inhibitor Screening Kit and human Plk-1 recombinant enzyme, and further synthesized the compounds to obtain Plk-1 As a result of examining the kinase inhibitory action, it was found that the heterocyclic compounds represented by the following formulas (1), (2) and (3) have an excellent Plk-1 inhibitory action.

  That is, the present invention relates to the general formula (1)

[Wherein the heterocyclic ring represented by A represents pyridine, pyrimidine, dihydropyrimidine, benzimidazole, benzothiazole, dihydrothiazole or thiazolidine;
The ring represented by B represents benzene or furan;
R ¹ represents a nitro group, an oxo group, a lower alkoxy group, a carboxyl group, a lower alkoxycarbonyl group, a lower alkanoyl group, an amino group, a thioxo group or a cyano group;
R ² represents a halogen atom, a lower alkoxy group, a lower alkoxycarbonyl group, a lower alkanoyl group or a cyano group;
X represents -S-, -O- or -NH-;
l represents an integer of 0 to 3 (however, when l is 2 or more, each R ² may be the same or different);
m represents an integer of 0 or 1;
n represents an integer of 0 to 3 (provided that when n represents 2 or more, each of R ¹ may be the same or different). ;
p shows the integer of 0-3. ]
The polo-like kinase-1 inhibitor which uses the heterocyclic compound represented by these, or its salt as an active ingredient is provided.

  Further, the present invention provides a compound represented by the general formula (2)

R ⁴ represents a phenyl group which may have a substituent, a 1-pyrazolyl group which may have a substituent, or a 4-morpholinyl group which may have a substituent. ]
The polo-like kinase-1 inhibitor which uses the heterocyclic compound represented by these, or its salt as an active ingredient is provided.

  In addition, the present invention provides a general formula (3)

[Wherein R ⁵ represents a phenethyl group which may have a substituent or a phenyl group which may have a substituent;
R ⁶ represents a hydrogen atom, R ⁷ represents a hydrogen atom or a cyano group, or R ⁶ and R ⁷ together form a 6-membered cyclic ether optionally substituted by an alkyl group;
R ⁸ represents a hydrogen atom, an amino group or a 4-morpholinyl group. ]
The polo-like kinase-1 inhibitor which uses the heterocyclic compound represented by these, or its salt as an active ingredient is provided.

  The present invention also provides a prophylactic / therapeutic agent for polo-like kinase-1 enhancing disease comprising the heterocyclic compound of the above formulas (1) to (3) or a salt thereof as an active ingredient.

Moreover, this invention provides the cancer metastasis inhibitor which uses the heterocyclic compound or its salt of said Formula (1)-(3) as an active ingredient.
Moreover, this invention provides the anticancer agent which uses the heterocyclic compound or its salt of said Formula (1)-(3) as an active ingredient.

  Moreover, this invention provides the therapeutic agent for rheumatoid arthritis which uses the heterocyclic compound or its salt of said Formula (1)-(3) as an active ingredient.

  Moreover, this invention provides the Alzheimer therapeutic agent which uses the heterocyclic compound or its salt of said Formula (1)-(3) as an active ingredient.

  Moreover, this invention provides the pharmaceutical composition which uses the heterocyclic compound or its salt of said Formula (1)-(3) as an active ingredient.

Moreover, this invention provides the polo-like kinase-1 inhibitor which uses the heterocyclic compound or its salt chosen from the following as an active ingredient.
N- [2- (4-fluorophenyl) ethyl] -2- (pyrimidin-2-ylsulfanyl) acetamide,
2- (pyridin-2-ylsulfanyl) -N- [2- (2,3,4-trimethoxyphenyl) ethyl] acetamide,
N- [2- (4-fluorophenyl) ethyl] -2- (5-nitropyridin-2-ylsulfanyl) acetamide,
N- [2- (4-fluorophenyl) ethyl] -2- (6-oxo-1,6-dihydropyrimidin-2-ylsulfanyl) acetamide,
N- [2- (4-fluorophenyl) ethyl] -2-[(6-methoxy-1H-benzimidazol-2-ylsulfanyl) acetamide,
N- [2- (4-fluorophenyl) ethyl] -2- (5-nitrobenzothiazol-2-ylsulfanyl) acetamide,
2-[[2- (4-fluorophenyl) ethylcarbamoyl] methylsulfanyl] nicotinic acid,
2-[[2- (4-fluorophenyl) ethylcarbamoyl] methylsulfanyl] nicotinic acid methyl ester,
2- (5-amino-1H-benzimidazol-2-ylsulfanyl) -N- [2- (4-fluorophenyl) ethyl] acetamide hydrochloride,
N- [2- (4-fluorophenyl) ethyl] -2- (2-pyrimidin-2-yloxy) acetamide,
N- [2- (4-fluorophenyl) ethyl] -2- (pyridin-2-yloxy) acetamide,
2- (4,5-dihydrothiazol-2-ylsulfanyl) -N- [2- (4-fluorophenyl) ethyl] acetamide,
N- [2- (4-fluorophenyl) ethyl] -2- (2-pyrimidin-2-ylamino) acetamide,
N- [2- (4-fluorophenyl) ethyl] -2- (2-thioxothiazolidin-3-yl) acetamide,
N- (4-acetylphenyl) -2- (6-amino-3,5-dicyano-2-pyridin-2-ylsulfanyl) acetamide,
2- (6-amino-3,5-dicyanopyridin-2-ylsulfanyl) -N- [2- (4-fluorophenyl) ethyl] acetamide,
2- (6-amino-3,5-dicyanopyridin-2-ylsulfanyl) -N-furan-2-ylmethylacetamide,
2- (6-amino-3,5-dicyanopyridin-2-ylsulfanyl) -N- (4-cyanophenyl) acetamide,
2-amino-6- (2-oxo-2-phenylethylsulfanyl) pyridine-3,5-dicarbonitrile,
1-[[(6-Amino-3,5-dicyano-2-pyridinyl) thio] acetyl] -3,5-dimethyl-1H-pyrazole
2-amino-6- (2-morpholin-4-ylethylsulfanyl) pyridine-3,5-dicarbonitrile,
3,6-diamino-5-cyanothieno [2,3-b] pyridine-2-carboxylic acid [2- (4-fluorophenyl) ethyl] amide,
N- (4-acetylphenyl) -1-amino-8,9-dihydro-8,8-dimethyl-5- (4-morpholinyl) -6H-pyrano [4,3-d] thieno [2,3-b ] Pyridine-2-carboxamide

The present invention also provides a heterocyclic compound selected from the following or a salt thereof.
2- (pyridin-2-ylsulfanyl) -N- [2- (2,3,4-trimethoxyphenyl) ethyl] acetamide,
N- [2- (4-fluorophenyl) ethyl] -2- (5-nitropyridin-2-ylsulfanyl) acetamide,
N- [2- (4-fluorophenyl) ethyl] -2- (6-oxo-1,6-dihydropyrimidin-2-ylsulfanyl) acetamide,
N- [2- (4-fluorophenyl) ethyl] -2-[(6-methoxy-1H-benzimidazol-2-ylsulfanyl) acetamide,
N- [2- (4-fluorophenyl) ethyl] -2- (5-nitrobenzothiazol-2-ylsulfanyl) acetamide,
2-[[2- (4-fluorophenyl) ethylcarbamoyl] methylsulfanyl] nicotinic acid,
2-[[2- (4-fluorophenyl) ethylcarbamoyl] methylsulfanyl] nicotinic acid methyl ester,
2- (5-amino-1H-benzimidazol-2-ylsulfanyl) -N- [2- (4-fluorophenyl) ethyl] acetamide hydrochloride,
N- [2- (4-fluorophenyl) ethyl] -2- (2-pyrimidin-2-yloxy) acetamide,
N- [2- (4-fluorophenyl) ethyl] -2- (pyridin-2-yloxy) acetamide,
2- (4,5-dihydrothiazol-2-ylsulfanyl) -N- [2- (4-fluorophenyl) ethyl] acetamide,
N- [2- (4-fluorophenyl) ethyl] -2- (2-pyrimidin-2-ylamino) acetamide,
N- [2- (4-Fluorophenyl) ethyl] -2- (2-thioxothiazolidin-3-yl) acetamide

  The compound of the present invention and a pharmaceutically acceptable salt thereof can be used for the prevention and treatment of diseases caused by cancer metastasis and proliferation, rheumatoid arthritis, and Alkheimer's enhanced Plk-1.

First, the compound represented by the general formula (1) will be described.
In the formula (1), examples of the heterocyclic ring represented by A (also referred to as heterocyclic ring A) include pyridine, pyrimidine, dihydropyrimidine, benzimidazole, benzothiazole, dihydrothiazole, and thiazolidine.
The heterocyclic ring A is a ring which may have one or a plurality of substituents R ¹ , and when there are a plurality of the substituents, each substituent may be the same or different.
Among the substituents R ¹ heterocyclic A may have a, which may have a substituent R ¹ 2-pyridyl group, which may have a substituent R ¹ 2-pyrimidyl group, substituents R ¹ may have a 1,6-dihydro-pyrimidin-2-yl group, which may have a substituent R ¹ 1H-benzimidazol-2-yl group, which may have a substituent R ¹ benzo thiazol-2-yl group, the substituent R ¹ may also be 4,5-dihydro-2-yl group having a or a substituent R ¹ may have a 4-thiazolidin-3-yl group is preferable .

Examples of the substituent R ¹ include a nitro group, an oxo group, a lower alkoxy group, a carboxyl group, a lower alkoxycarbonyl group, a lower alkanoyl group, an amino group, a thioxo group, and a cyano group.
Examples of the lower alkoxy group include an alkoxy group having 1 to 4 carbon atoms, and specific examples include a methoxy group, an ethoxy group, a propoxy group, and a butoxy group, and a methoxy group is particularly preferable.
Moreover, as the said lower alkoxycarbonyl group, a C1-C6 alkoxycarbonyl group is mentioned, Specifically, a methoxycarbonyl group, an ethoxycarbonyl group, etc. are mentioned.
Moreover, as a lower alkanoyl group, a C1-C6 alkanoyl group is mentioned, Specifically, a formyl group, an acetyl group, a propionyl group, a butyryl group, an isobutyryl group etc. are mentioned, An acetyl group is especially preferable.

  Among the heterocyclic rings A, in particular, 2-pyrimidine group, 2-pyridine group, 5-nitropyridin-2-yl group, 6-oxo-1,6-dihydropyrimidin-2-yl group, 6-methoxy-1H -Benzoimidazol-2-yl group, 5-nitrobenzothiazol-2-yl, 3-carboxy-2-pyridine group, 3-methoxycarbonyl-2-pyridine group, 5-amino-1H-benzimidazol-2-yl Group, 4,5-dihydrothiazol-2-yl, 2-thioxothiazolidin-3-yl group and 6-amino-3,5-dicyano-2-pyridin-2-yl group are preferable.

In the formula (1), examples of the ring represented by B (also referred to as ring B) include benzene and furan.
The ring B is a ring which may have one or a plurality of substituents R ² , and when there are a plurality of the substituents, the respective substituents may be the same or different.
Wherein among which may ring B substituted R ^2, phenyl group which may have a substituent R ² or a substituent may also 2-furyl group have R ² is preferable.

In the formula (1), examples of the substituent R ² include a halogen atom, a lower alkoxycarbonyl group, a lower alkoxy group, a lower alkanoyl group, and a cyano group, and specifically include a fluorine atom, a methoxy group, an acetyl group, a cyano group. Groups are more preferred.
Examples of the halogen atom include chlorine, bromine, fluorine, iodine and the like in addition to the fluorine.
The lower alkoxycarbonyl group, lower alkoxy group, and lower alkanoyl group are the same as defined above for R ¹ .

When the ring B is a phenyl group, it preferably has the substituent R ² at least at the 4-position of the phenyl group, and in particular, a 4-fluorophenyl group, a 2,3,4-trimethoxyphenyl group, 4 -Acetylphenyl group and 4-cyanophenyl group are preferable.

  In the formula (1), examples of X include a sulfur atom (—S—), an oxygen atom (—O—), and —NH—.

In the formula (1), l represents 0 or an integer of 1 to 3 (provided that when l represents 2 or more, each R ² may be the same or different).

  In the formula (1), m represents an integer of 0 or 1.

In the formula (1), n represents 0 or an integer of 1 to 3 (provided that when n represents 2 or more, each R ¹ may be the same or different).

  In the formula (1), p represents 0 or an integer of 1 to 3. p is preferably 0-2.

  In addition, in the general formula (1), a compound represented by the following general formula (4) is preferable.

[Wherein, A, R ¹ , R ² , X, l, m, n are the same as defined above. ]

  In the general formula (4), the case where m = 1 is particularly preferable.

  In the general formula (1), when p is 0 or 1, a compound represented by the following formula (1a) is preferable.

[Wherein p represents an integer of 0 or 1, and X, B, R ² and l are the same as defined above. ]

Next, the compound represented by the general formula (2) will be described.
In the formula (2), examples of R ³ include an oxygen atom (═O) and a hydrogen atom (—H).
In the formula (2), examples of R ⁴ include a phenyl group which may have a substituent, a 1-pyrazolyl group which may have a substituent, and a 4-morpholinyl group which may have a substituent. . These rings are rings which may have one or a plurality of substituents, and when there are a plurality of such substituents, each substituent may be the same or different.
Examples of the substituent for R ⁴ include a halogen atom, a cyano group, a lower alkyl group, and a lower alkanoyl group.
The halogen atom and lower alkanoyl group are the same as the definition of the substituent R ¹ .
Examples of the lower alkyl group include an alkyl group having 1 to 4 carbon atoms, and specific examples include a methyl group, an ethyl group, a propyl group, and a butyl group.
Of the substituents, an acetyl group, a fluorine atom, a cyano group, and a methyl group are preferable.
Of the R ⁴ , a phenyl group, a 3,5-dimethylpyrazolyl group or a 4-morpholinyl group is particularly preferable.

Next, the compound represented by the general formula (3) will be described.
In Formula (3), examples of R ⁵ include a phenethyl group that may have a substituent and a phenyl group that may have a substituent. These rings are rings which may have one or a plurality of substituents, and when there are a plurality of such substituents, each substituent may be the same or different.
Examples of the substituent for R ⁵ include a halogen atom and a lower alkoxycarbonyl group. The halogen atom has the same definition as the substituent R ² . The lower alkoxycarbonyl group has the same definition as the substituent R ¹ .
Of these substituents, a fluorine atom and an acetyl group are preferred.
Of the R ⁵ , 4-fluorophenethyl group or 4-acetylphenyl group is particularly preferable.

In the formula (3), when R ⁶ is a hydrogen atom, examples of R ⁷ include a hydrogen atom and a cyano group, or R ⁶ and R ⁷ together form a 6-membered ring which may be substituted with an alkyl group. A cyclic ether is formed. Examples of the ^6- membered cyclic ether which may be substituted by the alkyl group formed by combining R ⁶ and R ⁷ include 2,2-dimethyl-3,6-dihydro-2H-pyran.

In the formula (3), examples of R ⁸ include a hydrogen atom, an amino group, and a 4-morpholinyl group.

  In the present invention, the compounds of the above formulas (2) to (4) or pharmaceutically acceptable salts thereof are more preferable in that Plk-1 is strongly inhibited.

In the present invention, the following compounds or pharmaceutically acceptable salts thereof are more preferable in that Plk-1 is strongly inhibited.
N- [2- (4-fluorophenyl) ethyl] -2- (pyrimidin-2-ylsulfanyl) acetamide (hereinafter sometimes referred to as Compound 1),
2- (pyridin-2-ylsulfanyl) -N- [2- (2,3,4-trimethoxyphenyl) ethyl] acetamide (hereinafter sometimes referred to as Compound 2),
N- [2- (4-fluorophenyl) ethyl] -2- (5-nitropyridin-2-ylsulfanyl) acetamide (hereinafter sometimes referred to as Compound 4),
N- [2- (4-fluorophenyl) ethyl] -2- (6-oxo-1,6-dihydropyrimidin-2-ylsulfanyl) acetamide (hereinafter sometimes referred to as Compound 5),
N- [2- (4-fluorophenyl) ethyl] -2-[(6-methoxy-1H-benzimidazol-2-ylsulfanyl) acetamide (hereinafter sometimes referred to as Compound 6),
N- [2- (4-fluorophenyl) ethyl] -2- (5-nitrobenzothiazol-2-ylsulfanyl) acetamide (hereinafter sometimes referred to as Compound 7),
2-[[2- (4-fluorophenyl) ethylcarbamoyl] methylsulfanyl] nicotinic acid (hereinafter sometimes referred to as Compound 8),
2-[[2- (4-fluorophenyl) ethylcarbamoyl] methylsulfanyl] nicotinic acid methyl ester (hereinafter sometimes referred to as compound 9),
2- (5-amino-1H-benzimidazol-2-ylsulfanyl) -N- [2- (4-fluorophenyl) ethyl] acetamide hydrochloride (hereinafter sometimes referred to as Compound 10),
N- [2- (4-fluorophenyl) ethyl] -2- (2-pyrimidin-2-yloxy) acetamide (hereinafter sometimes referred to as Compound 11),
N- [2- (4-fluorophenyl) ethyl] -2- (pyridin-2-yloxy) acetamide (hereinafter sometimes referred to as Compound 12),
2- (4,5-dihydrothiazol-2-ylsulfanyl) -N- [2- (4-fluorophenyl) ethyl] acetamide (hereinafter sometimes referred to as Compound 13),
N- [2- (4-fluorophenyl) ethyl] -2- (2-pyrimidin-2-ylamino) acetamide (hereinafter sometimes referred to as Compound 17),
N- [2- (4-fluorophenyl) ethyl] -2- (2-thioxothiazolidin-3-yl) acetamide (hereinafter sometimes referred to as Compound 14),
N- (4-acetylphenyl) -2- (6-amino-3,5-dicyano-2-pyridin-2-ylsulfanyl) acetamide (hereinafter sometimes referred to as compound 18),
2- (6-amino-3,5-dicyanopyridin-2-ylsulfanyl) -N- [2- (4-fluorophenyl) ethyl] acetamide (hereinafter sometimes referred to as Compound 19),
2- (6-amino-3,5-dicyanopyridin-2-ylsulfanyl) -N-furan-2-ylmethylacetamide (hereinafter sometimes referred to as Compound 20),
2- (6-amino-3,5-dicyanopyridin-2-ylsulfanyl) -N- (4-cyanophenyl) acetamide (hereinafter sometimes referred to as Compound 21),
2-amino-6- (2-oxo-2-phenylethylsulfanyl) pyridine-3,5-dicarbonitrile (hereinafter sometimes referred to as compound 22),
1-[[(6-amino-3,5-dicyano-2-pyridinyl) thio] acetyl] -3,5-dimethyl-1H-pyrazole (hereinafter sometimes referred to as Compound 23),
2-amino-6- (2-morpholin-4-ylethylsulfanyl) pyridine-3,5-dicarbonitrile (hereinafter sometimes referred to as compound 24),
3,6-diamino-5-cyanothieno [2,3-b] pyridine-2-carboxylic acid [2- (4-fluorophenyl) ethyl] amide (hereinafter sometimes referred to as compound 25),
N- (4-acetylphenyl) -1-amino-8,9-dihydro-8,8-dimethyl-5- (4-morpholinyl) -6H-pyrano [4,3-d] thieno [2,3-b ] Pyridine-2-carboxamide (hereinafter sometimes referred to as Compound 26)

In the present invention, the following compounds or pharmaceutically acceptable salts thereof are particularly preferable in that Plk-1 is strongly inhibited.
2- (pyridin-2-ylsulfanyl) -N- [2- (2,3,4-trimethoxyphenyl) ethyl] acetamide (compound 2),
N- [2- (4-fluorophenyl) ethyl] -2- (5-nitropyridin-2-ylsulfanyl) acetamide (compound 4),
N- [2- (4-fluorophenyl) ethyl] -2- (6-oxo-1,6-dihydropyrimidin-2-ylsulfanyl) acetamide (compound 5),
N- [2- (4-fluorophenyl) ethyl] -2-[(6-methoxy-1H-benzimidazol-2-ylsulfanyl) acetamide (Compound 6),
N- [2- (4-fluorophenyl) ethyl] -2- (5-nitrobenzothiazol-2-ylsulfanyl) acetamide (Compound 7),
2-[[2- (4-fluorophenyl) ethylcarbamoyl] methylsulfanyl] nicotinic acid (Compound 8),
2-[[2- (4-fluorophenyl) ethylcarbamoyl] methylsulfanyl] nicotinic acid methyl ester (compound 9),
2- (5-amino-1H-benzimidazol-2-ylsulfanyl) -N- [2- (4-fluorophenyl) ethyl] acetamide hydrochloride (compound 10),
N- [2- (4-fluorophenyl) ethyl] -2- (2-pyrimidin-2-yloxy) acetamide (Compound 11),
N- [2- (4-fluorophenyl) ethyl] -2- (pyridin-2-yloxy) acetamide (Compound 12),
2- (4,5-dihydrothiazol-2-ylsulfanyl) -N- [2- (4-fluorophenyl) ethyl] acetamide (compound 13),
N- [2- (4-fluorophenyl) ethyl] -2- (2-pyrimidin-2-ylamino) acetamide (Compound 17),
N- [2- (4-Fluorophenyl) ethyl] -2- (2-thioxothiazolidin-3-yl) acetamide (Compound 14)

  In the present invention, pharmaceutically acceptable salts include hydrochloride, sulfate, citrate, tartrate, acetate, methanesulfonate, phosphate, oxalate, benzoate, trifluoroacetic acid. Acid addition salts such as salts, maleates, fumarate, lactates, bromates, iodates, succinates and glutarates, and metal salts such as lithium, sodium, potassium and magnesium salts Is mentioned.

In the present invention, the “polo-like kinase-1 inhibitor” is not particularly limited as long as it has an inhibitory action on polo-like kinase-1, but is determined by the method described later (Example 17). The inhibitory action (IC ₅₀ ) on polo-like kinase-1 (concentration of the compound of the present invention that inhibits enzyme activity 50%) is preferably 100 μM or less, more preferably 50 μM or less, and particularly preferably 20 μM or less. Preferably, 10 μM or less is most preferable.

In addition, as the “polo-like kinase-1 inhibitor”, among various serine-threonine kinases, it has an inhibitory action on polo-like kinase-1 and other serine-threonine kinases (for example, Aurora A, AKT, etc.) ) Is preferable in terms of reducing side effects and toxicity, and more specifically, an inhibitory effect on Aurora A and AKT determined by the methods described below (Examples 18 and 19). (IC ₅₀ ) (concentration of the compound of the present invention that inhibits enzyme activity by 50%) is more preferably 70 μM or more, particularly preferably 100 μM or more.
Furthermore, when the compound of the present invention is used as an anticancer agent, a compound that exhibits a strong inhibitory action on the growth of cancer cells is preferred, and more specifically, by the method described below (Example 20). The inhibitory action (IC ₅₀ ) (concentration of the compound of the present invention that inhibits cell proliferation by 50%) on the proliferation of non-small cell lung cancer A549 cells is more preferably 20 μM or less, and particularly preferably 10 μM or less. .

  The compound of the present invention can be produced, for example, according to the following reaction formula.

[Wherein, A, B, R ¹ , R ² , X, l, m, n, and p are the same as described above. ]

That is, the heterocyclic compound of the general formula (1) or a salt thereof can be obtained by condensing the amine compound of the general formula (5) and the carboxylic acid of the general formula (6).
Moreover, the heterocyclic compound or its salt of General formula (1) is obtained by condensing the compound of General formula (7), and the compound of General formula (8).
Moreover, the heterocyclic compound or its salt of General formula (1) is obtained by condensing the compound of General formula (9) and the compound of General formula (10).
Here, the condensation reaction is preferably performed in the presence of a base such as potassium carbonate, sodium carbonate, sodium hydroxide or triethylamine.

  The compounds of the present invention inhibit Plk-1 against diseases caused by increased Plk-1 by administering an effective amount in a pharmaceutically acceptable carrier or alone to a patient, thereby Used for disease prevention, treatment and symptom improvement. The compound of the present invention may be appropriately formulated to increase the efficiency of transport of the compound to tumor tissue, synovial tissue or brain tissue. Here, examples of the disease caused by the increase in Plk-1 include cancer metastasis and proliferation, rheumatoid arthritis, and Alzheimer.

  Of the compounds of the present invention, one of them may be used alone, or a plurality of them may be used in combination. Furthermore, it may be used in combination with other therapeutically advantageous compounds, and the mechanism of action of the other compounds may be the same as or different from the compounds of the present invention. The preferred form of systemic administration of a pharmaceutical composition comprising a compound of the present invention is injection, oral, and in the case of injection, especially intravenous injection. Other injection routes such as subcutaneous, intramuscular or intraperitoneal can also be used. Another means for systemic administration is transmucosal or transdermal administration using penetrants such as bile salts or fucdic acid or other surfactants. Furthermore, oral administration is possible by enteric formulation or capsule formulation. Administration of these pharmaceutical compositions may be local, and may be in the form of a salve, pasta, gel or the like.

  There is no strict limitation on the dosage when using the heterocyclic compound or its salt which is an active ingredient of the Plk-1 inhibitor of the present invention. The effect obtained varies depending on various use modes such as the subject and applicable disease, and therefore it is desirable to set the dose as appropriate. The preferred dose is 1 mg to 10 g, more preferably 10 mg to 1 g per day. It is.

  Examples of such preparations include solid agents such as tablets, granules, powders and capsules, solutions such as solutions, suspensions and emulsions, and freeze-dried agents. These preparations can be prepared by conventional means on the preparation. Examples of the non-toxic pharmaceutical carrier include starch, dextrin, fatty acid glyceride, polyethylene glycol, hydroxyethyl starch, ethylene glycol, polyoxyethylene sorbitan fatty acid ester, amino acid, gelatin, albumin, water, and physiological saline. It is done. In addition, conventional additives such as stabilizers, wetting agents, emulsifiers, binders, isotonic agents, excipients and the like can be appropriately added as necessary.

  Moreover, the Plk-1 inhibitor of the present invention can be used not only as a pharmaceutical preparation as described above but also as a food or drink. In this case, the heterocyclic compound of the present invention or a salt thereof may be contained in the food or drink as it is or with various nutritional components added. This food and drink can be used as a health food or food material useful for the improvement and prevention of cancer metastasis and proliferation, rheumatoid arthritis, Alzheimer, etc., and these foods and drinks or containers thereof have the above-mentioned effects An indication to that effect may be attached. Specifically, when the Plk-1 inhibitor of the present invention is blended in a food or drink, an additive that can be used as a food or drink is appropriately used, and a form suitable for food using conventional means, for example, granular, It may be formed into granules, tablets, capsules, pastes, etc., and various foods such as processed meat products such as ham and sausage, processed fish products such as kamaboko and chikuwa, bread, confectionery, butter, powdered milk, fermented food and drink It may be used by adding to a product, or may be used by adding to beverages such as water, fruit juice, milk, soft drinks and tea beverages. The food and drink includes animal feed.

  Furthermore, fermented milk products such as fermented milk, lactic acid bacteria beverages, fermented soy milk, fermented fruit juice, fermented plant liquid, etc. containing a heterocyclic compound or salt thereof as an active ingredient are suitably used as food and drink. These fermented milk foods and drinks can be produced according to a conventional method. For example, fermented milk is obtained by inoculating and cultivating lactic acid bacteria or bifidobacteria in a sterilized milk medium, and homogenizing it to obtain a fermented milk base. Subsequently, a separately prepared syrup solution and the heterocyclic compound of the present invention or a salt thereof are added and mixed, homogenized with a homogenizer or the like, and further added with flavor to obtain a final product. The fermented milk thus obtained can be made into a product of any form such as a plain type, a soft type, a fruit flavor type, a solid form, and a liquid form.

  Moreover, the Plk-1 inhibitor of this invention is applicable to all mammals including a human.

  Hereinafter, the content of the present invention will be described in more detail with reference to examples, but the present invention is not limited by these.

Example 1
N- [2- (4-Fluorophenyl) ethyl] -2- (pyrimidin-2-ylsulfanyl) acetamide (Compound 1)
To a solution of bromoacetic acid (290 mg, 2.08 mmol) in dimethylformamide (10 mL) were added 2-mercaptopyrimidine (234 mg, 2.08 mmol) and triethylamine (0.6 mL, 4.17 mmol), and the mixture was stirred at 95 ° C. for 3 hours. Thereafter, the temperature was returned to room temperature, and 4-fluorophenethylamine (290 mg, 2.08 mmol) was added. Then 1-ethyl-3- (3-dimethylaminopropyl) carbodiimide hydrochloride (400 mg, 2.08 mmol), triethylamine (0.3 mL, 2.08 mmol) and dimethylaminopyridine (25 mg, 0.2 mmol) were added, and Stir for hours. The reaction mixture was concentrated under reduced pressure, and the residue was purified by silica gel column chromatography (chloroform containing 3% methanol) to obtain the title compound (140 mg, 23%).
¹ H-NMR (CDCl ₃ ) δ: 2.73 (2H, t, J = 7Hz), 3.49 (2H, q like), 3.73 (2H, s), 6.78-6.83 (2H, m), 6.98-7.03 (4H , m), 8.41 (2H, d, J = 5Hz).
MS m / z: 291 (M +).

Example 2
2- (Pyridin-2-ylsulfanyl) -N- [2- (2,3,4-trimethoxyphenyl) ethyl] acetamide (Compound 2)
To a solution of bromoacetic acid (290 mg, 2.08 mmol) in dimethylformamide (10 mL) were added 2-mercaptopyridine (232 mg, 2.08 mmol) and triethylamine (0.6 mL, 4.17 mmol), and the mixture was stirred at 95 ° C. for 3 hours. Then, the temperature was returned to room temperature, and 2,3,4-trimethoxyphenethylamine (440 mg, 2.08 mmol) was added. Then 1-ethyl-3- (3-dimethylaminopropyl) carbodiimide hydrochloride (400 mg, 2.08 mmol), triethylamine (0.3 mL, 2.08 mmol) and dimethylaminopyridine (25 mg, 0.2 mmol) were added, and Stir for hours. The reaction mixture was concentrated under reduced pressure, and the residue was purified by silica gel column chromatography (chloroform) to obtain the title compound (639 mg, 90%).
¹ H-NMR (CDCl ₃ ) δ: 2.69 (2H, t, J = 7Hz), 3.45 (2H, q like), 3.79 (2H, s), 3.80 (3H, s), 3.82 (3H, s), 3.83 (3H, s), 6.43 (1H, d, J = 9Hz), 6.70 (1H, d, J = 9Hz), 6.98-7.02 (1H, m), 7.19 (1H, d, J = 8Hz), 7.42 -7.53 (2H, m), 8.23-8.25 (2H, m).
MS m / z: 362 (M +).

Example 3
2-Bromo-N- [2- (4-fluorophenyl) ethyl] acetamide (Compound 3)
4-Fluorophenethylamine (2.64 mL, 20 mmol) was added to a dimethylformamide solution (70 mL) of bromoacetic acid (2.78 g, 20 mmol), and then 1-ethyl-3- (3-dimethylaminopropyl) carbodiimide hydrochloride ( 3.84 g, 20 mmol) and dimethylaminopyridine (250 mg, 2 mmol) were added and stirred at 0-10 ° C. for 5 days. The reaction mixture was concentrated under reduced pressure, and the residue was purified by silica gel column chromatography (chloroform) to obtain the title compound (2.94 g, 57%).
¹ H-NMR (CDCl ₃ ) δ: 2.82 (2H, t, J = 7Hz), 3.54 (2H, q like), 4.03 (2H, s), 6.58 (1H, br-s), 7.01 (2H, t , J = 9Hz), 7.14-7.26 (2H, m).

Example 4
N- [2- (4-Fluorophenyl) ethyl] -2- (5-nitropyridin-2-ylsulfanyl) acetamide (Compound 4)
To a solution of compound 3 (107 mg, 0.41 mmol) in dimethylformamide (2 mL) was added 5-nitropyridine-2-thiol (64 mg, 0.41 mmol) and triethylamine (114 μL, 0.82 mmol), and 1 Stir for hours. Dimethylformamide was distilled off under reduced pressure, and the residue was dissolved in chloroform and washed with a saturated aqueous sodium hydrogen carbonate solution and then with water. The chloroform layer was dried over anhydrous sodium sulfate and concentrated to dryness under reduced pressure to obtain a brown oily residue. The residue was crystallized from n-hexane-chloroform, and the crystals were collected by filtration to give the title compound (130 mg, 95%).
¹ H-NMR (CDCl ₃ ) δ: 2.73 (2H, t, J = 7Hz), 3.50 (2H, q like), 3.79 (2H, s), 6.70-6.87 (3H, m), 6.94-7.05 (2H , m), 7.30 (1H, d, J = 9Hz), 8.25 (1H, dd, J = 3, 9Hz), 8.93 (1H, d, J = 3Hz).
MS m / z: 335 (M +).

Example 5
N- [2- (4-Fluorophenyl) ethyl] -2- (6-oxo-1,6-dihydropyrimidin-2-ylsulfanyl) acetamide (Compound 5)
2 mercapto-3H-pyrimidin-4-one (49 mg, 0.38 mmol) and triethylamine (110 μL, 0.79 mmol) were added to a solution of compound 3 (100 mg, 0.38 mmol) in dimethylformamide (2 mL) at 90 ° C. For 3 hours. Dimethylformamide was distilled off under reduced pressure, and the residue was dissolved in chloroform and washed with water. The chloroform layer was dried over anhydrous sodium sulfate and concentrated to dryness under reduced pressure to obtain a white solid. This was pulverized by irradiating with ultrasonic waves in a mixture of n-hexane-chloroform, and the crystals were collected by filtration to obtain the title compound (88 mg, 75%).
¹ H-NMR (CDCl ₃ ) δ: 2.69 (2H, t, J = 7Hz), 3.28 (2H, q like), 3.82 (2H, s), 6.12 (1H, br-s), 7.02-7.15 (2H , m), 7.17-7.27 (2H, m), 7.82 (1H, br-s), 8.19 (1H, br-t, J = 5Hz), 12.75 (1H, br).
MS m / z: 307 (M +).

Example 6
N- [2- (4-Fluorophenyl) ethyl] -2-[(6-methoxy-1H-benzimidazol-2-ylsulfanyl) acetamide (Compound 6)
To a solution of compound 3 (100 mg, 0.38 mmol) in dimethylformamide (2 mL) was added 6-methoxy-1H-benzimidazol-2-thiol (69 mg, 0.38 mmol) and triethylamine (110 μL, 0.79 mmol). The mixture was stirred at 90 ° C. for 3 hours. Dimethylformamide was distilled off under reduced pressure, and the residue was dissolved in chloroform and washed with water. The chloroform layer was dried over anhydrous sodium sulfate and concentrated to dryness under reduced pressure to give a pale orange viscous oil. This viscous oil was purified by silica gel column chromatography (n-hexane-containing ethyl acetate; n-hexane content 50% → 33%) to obtain the title compound (121 mg, 88%).
¹ H-NMR (CDCl ₃ ) δ: 2.76 (2H, t, J = 7Hz), 3.52 (2H, q like), 3.72 (2H, s), 3.83 (3H, s), 6.73-6.87 (3H, m ), 6.93 (1H, br-s), 6.98-.706 (2H, m), 7.32 (1H, br-d, J = 8Hz), 7.85 (1H, br-t like).
MS m / z: 359 (M +).

Example 7
N- [2- (4-Fluorophenyl) ethyl] -2- (5-nitrobenzothiazol-2-ylsulfanyl) acetamide (Compound 7)
To a solution of compound 3 (100 mg, 0.38 mmol) in dimethylformamide (2 mL) was added 6-nitro-2-mercaptobenzothiazole (82 mg, 0.38 mmol) and triethylamine (110 μL, 0.769 mmol), and 2 at 90 ° C. Stir for hours. Dimethylformamide was distilled off under reduced pressure, and the residue was dissolved in chloroform and washed with 0.1 mol / L sodium hydroxide and water. The chloroform layer was dried over anhydrous sodium sulfate and then concentrated to dryness under reduced pressure to obtain an oily residue. This was crystallized from n-hexane-chloroform, and the crystals were collected by filtration to give the title compound (129 mg, 86%).
¹ H-NMR (CDCl ₃ ) δ: 2.75 (2H, t, J = 7Hz), 3.55 (2H, q like), 3.69 (2H, s), 6.64-6.70 (2H, m), 6.94-6.99 (2H , m), 7.02 (1H, br-s), 7.67 (1H, d, J = 9Hz), 8.30 (1H, dd, J = 9, 2Hz,), 8.70 (1H, d, J = 2Hz).
MS m / z: 391 (M +).

Example 8
2-[[2- (4-Fluorophenyl) ethylcarbamoyl] methylsulfanyl] nicotinic acid (Compound 8)
2-mercaptonicotinic acid (60 mg, 0.38 mmol) and triethylamine (176 μL, 1.27 mmol) were added to a solution of compound 3 (130 mg, 0.50 mmol) in dimethylformamide (2 mL), and the mixture was stirred at 90 ° C. for 2 hours. . Dimethylformamide was distilled off under reduced pressure to obtain a residue. This was purified by silica gel column chromatography (methanol-containing chloroform; methanol content 2% → 5%) to obtain the title compound (73 mg, 57%).
¹ H-NMR (DMSO-d ₆ ) δ: 2.67 (2H, t, J = 7Hz), 3.27 (2H, q like), 3.77 (2H, s), 7.00-7.10 (2H, m), 7.14-7.30 (3H, m), 8.04 (1H, br-t, J = 5Hz), 8.23 (1H, m), 8.55 (1H, m), 13.50 (1H, br-s).
MS m / z: 334 (M +).

Example 9
2-[[2- (4-Fluorophenyl) ethylcarbamoyl] methylsulfanyl] nicotinic acid methyl ester (Compound 9)
Compound 8 (35 mg, 0.11 mmol) was dissolved in a mixture of chloroform (0.8 mL) -methanol (0.2 mL) and stirred at room temperature with trimethylsilyldiazomethane (0.5 mL [ca.10% hexane solution, ca.0.60 mol / L). ]) Was added and stirred for 30 minutes. The solvent was distilled off under reduced pressure to obtain a viscous oil. This was crystallized from an n-hexane-chloroform mixture, and the crystals were collected by filtration to give the title compound (25 mg, 70%).
¹ H-NMR (CDCl ₃ ) δ: 2.66 (2H, t, J = 7Hz), 3.44 (2H, q like), 3.77 (2H, s), 3.94 (3H, s), 6.69-6.80 (2H, m ), 6.83-6.99 (3H, m), 7.09 (1H, dd, J = 5, 8Hz), 8.24 (1H, dd, J = 2, 8Hz), 8.33 (1H, dd, J = 2, 5Hz).
MS m / z: 348 (M +).

Example 10
2- (5-Amino-1H-benzimidazol-2-ylsulfanyl) -N- [2- (4-fluorophenyl) ethyl] acetamide hydrochloride (Compound 10)
To a solution of compound 3 (200 mg, 0.77 mmol) in dimethylformamide (4 mL), 5-amino-2-mercaptobenzimidazole (128 mg, 0.77 mmol) and triethylamine (220 μL, 1.53 mmol) were added, and the mixture was heated at 80 ° C. for 4 hours. Stir for hours. Dimethylformamide was distilled off under reduced pressure, and the residue was dissolved in chloroform and washed with 0.1 mol / L sodium hydroxide and water. The chloroform layer was dried over anhydrous sodium sulfate and then concentrated to dryness under reduced pressure to obtain an oily residue. This was purified by silica gel column chromatography (chloroform containing 3% methanol). Then, a 4 mol / L hydrochloric acid-dioxane solution (0.057 mL) was added to a dioxane (1 mL) solution of this at room temperature. Hexane (40 mL) was added to this mixture, and only the supernatant was removed. This operation was repeated 5 times, and then the crystals were collected by filtration to obtain the title compound (63 mg, 23%).
¹ H-NMR (CDCl ₃ ) δ: 2.65 (2H, t, J = 7Hz), 3.27-3.33 (2H, m), 3.98 (2H, s), 6.97-7.04 (4H, m), 7.15-7.21 ( 3H, m), 7.42 (1H, d, J = 9Hz), 8.33-8.40 (1H, m).
MS m / z: 344 (M +).

Example 11
N- [2- (4-Fluorophenyl) ethyl] -2- (2-pyrimidin-2-yloxy) acetamide (Compound 11)
2-hydroxypyridine hydrochloride (51 mg, 0.38 mmol) and potassium carbonate (210 mg, 1.52 mmol) were added to a solution of compound 3 (100 mg, 0.38 mmol) in dimethylformamide (5 mL), and the mixture was stirred at room temperature for 17 hours. did. Dimethylformamide was distilled off under reduced pressure, chloroform and water were added to the residue, and the mixture was shaken well. The chloroform layer was dried over anhydrous sodium sulfate and then concentrated to dryness under reduced pressure to obtain an oily residue. This was purified by silica gel column chromatography (methanol-containing chloroform; methanol content 1.5% ⇒3%) to obtain the title compound (67 mg, 64%).
¹ H-NMR (DMSO-d ₆ ) δ: 2.71 (2H, t, J = 7Hz), 3.27-3.35 (2H, m), 4.46 (2H, s), 6.42 (1H, dd, J = 4, 6Hz ), 7.07-7.12 (2H, m), 7.23-7.27 (2H, m), 8.06 (1H, dd, J = 3, 7Hz), 8.29-8.32 (1H, m), 8.54 (1H, dd, J = 3, 4Hz).
MS m / z: 275 (M +).

Example 12
N- [2- (4-Fluorophenyl) ethyl] -2- (pyridin-2-yloxy) acetamide (Compound 12)
Compound 3 (100 mg, 0.38 mmol) and 2-hydroxypyrimidine (37 mg, 0.38 mmol) were used for reaction and post-treatment in the same manner as in Example 11 to obtain the title compound (103 mg, 97%).
¹ H-NMR (CDCl ₃ ) δ: 2.75 (2H, t, J = 7Hz), 3.45 (2H, q like), 4.49 (2H, s), 6.26 (1H, dd, J = 2, 7Hz), 6.60 (1H, d, J = 9Hz), 6.89-6.94 (2H, m), 6.96-7.09 (3H, m), 7.36 (1H, dd, J = 2, 7Hz), 7.39-7.43 (1H, m).
MS m / z: 274 (M +).

Example 13
2- (4,5-dihydrothiazol-2-ylsulfanyl) -N- [2- (4-fluorophenyl) ethyl] acetamide (compound 13) and N- [2- (4-fluorophenyl) ethyl] -2 -(2-Thioxothiazolidin-3-yl) acetamide (Compound 14)
2-mercaptothiazoline (46 mg, 0.38 mmol) and potassium carbonate (210 mg, 1.52 mmol) were added to a solution of compound 3 (100 mg, 0.38 mmol) in dimethylformamide (4 mL), and the mixture was stirred at room temperature for 17 hours. Dimethylformamide was distilled off under reduced pressure, chloroform and water were added to the residue, and the mixture was shaken well. The chloroform layer was dried over anhydrous sodium sulfate and then concentrated to dryness under reduced pressure to obtain an oily residue. Two components of this were purified by silica gel column chromatography (chloroform) to obtain Compound 13 (70 mg, 62%) and Compound 14 (4 mg, 5%).
Compound 13
¹ H-NMR (CDCl ₃ ) δ: 2.77 (2H, t, J = 7Hz), 3.37 (2H, t, J = 8Hz), 3.48 (2H, q like), 3.61 (2H, s), 3.92 (2H , t, J = 8Hz), 6.97-7.04 (2H, m), 7.12-7.18 (2H, m), 7.38 (1H, br-s).
MS m / z: 298 (M +).
Compound 14
¹ H-NMR (CDCl ₃ ) δ: 2.79 (2H, t, J = 7Hz), 3.27 (2H, t, J = 8Hz), 3.52 (2H, q like), 4.05 (2H, t, J = 8Hz) , 4.36 (2H, s), 6.97-7.03 (2H, m), 7.14-7.18 (2H, m).
MS m / z: 298 (M +).

Example 14
[[2- (4-Fluorophenyl) ethylcarbamoyl] -methyl] -carbamic acid tert-butyl ester (compound 15)
To a solution of N-Boc-glycine (1.75 g, 10 mmol) in dimethylformamide (40 mL) was added 4-fluorophenethylamine (1.31 mL, 10 mmol), followed by 1-ethyl-3- (3-dimethylaminopropyl) carbodiimide. Hydrochloride (1.91 g, 10 mmol) and dimethylaminopyridine (122 mg, 1 mmol) were added, and the mixture was stirred at room temperature for 20 hours. The reaction mixture was concentrated under reduced pressure, and the residue was purified by silica gel column chromatography (methanol-containing chloroform; methanol content 1% ⇒2%) to obtain the title compound (2.35 g, 80%).
¹ H-NMR (CDCl ₃ ) δ: 1.43 (9H, s), 2.79 (2H, t, J = 7Hz), 3.50 (2H, q like), 3.74 (2H, d, J = 6Hz), 5.09 (1H , br-s), 6.17 (1H, br-s), 6.99 (2H, t, J = 9Hz), 7.15 (2H, dd, J = 6, 9Hz).

Example 15
2-Amino-N- [2- (4-fluorophenyl) ethyl] acetamide (Compound 16)
Trifluoroacetic acid (2 mL, 26 mmol) was added to a dichloromethane solution (3 mL) of compound 15 (400 mg, 1.35 mmol), and the mixture was stirred at 0 ° C. for 1 hour. The reaction mixture is slowly poured into saturated sodium bicarbonate solution to make it alkaline. Chloroform was added to this mixture and shaken well. The chloroform layer was dried over anhydrous sodium sulfate and then concentrated to dryness under reduced pressure to obtain the title compound (120 mg, 45%).
¹ H-NMR (CDCl ₃ ) δ: 1.46 (2H, br-s), 2.81 (2H, t, J = 7Hz), 3.32 (2H, s), 3.52 (2H, q like), 6.97-7.01 (2H , m), 7.17 (2H, dd, J = 5, 9Hz), 7.31 (1H, br-s).

Example 16
N- [2- (4-Fluorophenyl) ethyl] -2- (2-pyrimidin-2-ylamino) acetamide (Compound 17)
2-Bromopyrimidine (61 mg, 0.38 mmol) and potassium carbonate (80 mg, 0.57 mmol) were added to a solution of compound 16 (75 mg, 0.38 mmol) in dimethylformamide (2 mL), and the mixture was stirred at 150 ° C. for 2 hours. . Dimethylformamide was distilled off under reduced pressure, chloroform and water were added to the residue, and the mixture was shaken well. The chloroform layer was dried over anhydrous sodium sulfate and then concentrated to dryness under reduced pressure to obtain a residue. This was purified by silica gel column chromatography (chloroform containing 1% methanol) to obtain the title compound (87 mg, 84%).
¹ H-NMR (CDCl ₃ ) δ: 2.76 (2H, t, J = 7Hz), 3.51 (2H, q like), 4.03 (2H, d, J = 6Hz), 5.56 (1H, br-s), 6.36 (1H, br-s), 6.65 (1H, t, J = 5Hz), 6.89 (2H, t, J = 9Hz), 7.05 (1H, dd, J = 6, 9Hz), 8.29 (2H, d, J = 5Hz).
MS m / z: 274 (M +).

Example 17
Inhibitory Action on Plk-1 Activity Using the compounds shown in Table 1, the in vitro Plk-1 inhibitory action of the heterocyclic compounds of the present invention was examined. Compounds 1, 4, and 13 were prepared by the methods shown in the above-mentioned Examples, respectively, Compounds 18 and 19 were from Akos Consulting & Solutions, Compound 20 was from Vitas-M Laboratory, and Compounds 21 to 24 were ART-CHEM. Compounds 25 and 26 were obtained from Pharmeks, respectively. Plk-1 activity was measured using the Polo-like kinase 1 Assay / Inhibitor Screening Kit and human Plk-1 recombinase (CycLex Co., Ltd.) with modifications to the attached operating procedure. The enzyme amount during the enzyme reaction was 0.2 mU / 100 μl, the ATP concentration was 1 μM, and the enzyme reaction time was 20 minutes. The test compound was used by dissolving in a DMSO solution, and the final concentration of DMSO in the enzyme reaction solution was 1%. The Plk-1 inhibitory action of the compounds of the present invention is shown in Table 1 as IC ₅₀ values (concentrations of heterocyclic compounds that inhibit enzyme activity 50%). As a result, the heterocyclic compound of the present invention showed a strong inhibitory action on Plk-1 activity.

Example 18
Action on Aurora A activity Using the compounds shown in Table 2, the effect on Aurora A belonging to the same serine / threonine kinase as Plk-1 was investigated for the purpose of examining the specificity of the heterocyclic compound of the present invention for Plk-1. investigated. Aurora A activity was measured using the Aurora-A kinase Assay / Inhibitor Screening Kit and human Aurora A recombinase (CycLex Co., Ltd.) with modifications to the attached operating procedure. The enzyme amount during the enzyme reaction was 20 mU / 100 μl, the ATP concentration was 10 μM, and the enzyme reaction time was 20 minutes. The test compound was used by dissolving in a DMSO solution, and the final concentration of DMSO in the enzyme reaction solution was 1%. The Aurora A inhibitory action of the compounds of the present invention is shown in Table 2 as IC ₅₀ values (concentrations of heterocyclic compounds that inhibit enzyme activity 50%). As a result, the heterocyclic compound of the present invention showed no Aurora A inhibitory action even at 100 μM, indicating high selectivity for Plk-1.

Example 19
Action on AKT activity Using the compounds shown in Table 3, the action on the AKT belonging to the same serine / threonine kinase as Plk-1 was examined for the purpose of examining the specificity of the heterocyclic compound of the present invention for Plk-1. . AKT activity was measured using the AKT / PKB kinase Assay / Inhibitor Screening Kit and human AKT 1 recombinant enzyme (CycLex Co., Ltd.) with modifications to the attached operating procedure. The enzyme amount during the enzyme reaction was 15 mU / 100 μl, the ATP concentration was 50 μM, and the enzyme reaction time was 90 minutes. The test compound was used by dissolving in a DMSO solution, and the final concentration of DMSO in the enzyme reaction solution was 1%. The AKT inhibitory action of the compounds of the present invention is shown in Table 3 as IC ₅₀ values (concentrations of heterocyclic compounds that inhibit enzyme activity 50%). As a result, the AKT inhibitory action of the heterocyclic compound of the present invention was weaker than the inhibitory action against Plk-1, and it was shown to have a high selectivity for Plk-1.

Example 20
Inhibitory effect on proliferation of cancer cells The effect of the heterocyclic compound of the present invention on the proliferation of non-small cell lung cancer A549 cells was examined. A549 cells were suspended in RPMI1640 medium (10% FBS / RPMI1640) containing 10% FBS, 100 U / mL penicillin and 100 μg / mL streptomycin, seeded in a 96-well microplate at 5% CO ₂ at 37 ° C. The cells were cultured (200 cells / 50 μl / well). After overnight culture, 50 μl of a medium in which the compound of the present invention was dissolved was added, and further cultured for 4 days. After culturing, the number of viable cells was measured using TetraColor ONE (Seikagaku Corporation) according to the attached operation procedure. The test compound was used after being dissolved in a DMSO solution, and the final concentration of DMSO in the medium was 0.1%. The results were expressed as the concentration of the test compound that suppressed cell growth by 50% (IC ₅₀ ). As a result, as shown in Table 4, the heterocyclic compound of the present invention showed a strong inhibitory action on the proliferation of A549 cells.

Example 21
Inhibitory effect on dephosphorylation of phosphorylated Cdc2 A549 cells for the purpose of confirming that the heterocyclic compound of the present invention, which showed an inhibitory action on proliferation of cancer cells, targets intracellular Plk-1. The effect of phosphorylated Cdc2 on the dephosphorylation in the rat was examined by Western blotting. The cells were suspended in 10% FBS / RPMI1640, seeded in a 10 cm dish, and cultured at 37 ° C. with 5% CO ₂ . After the cells grew to 50% confluence, the compound of the present invention was added and cultured for 24 hours. After that, the cells were washed twice with PBS, solubilized solution (10 mM Tris-HCl, pH 7.4, 0.1% NP-40, 0.1% sodium deoxycholate, 0.1% SDS, 0.15M NaCl, 1 mM EDTA, 10 μg / ml aprotinin) was added to lyse the cells. After adding trichloroacetic acid (final concentration 3.3%) to this lysate, centrifuge at 12,500 xg for 1 minute, wash the resulting precipitate with ether, and follow Laemmli's method (Nature, 1970, 227: 680-685) Electrophoresis was performed. After completion of electrophoresis, the protein in the acrylamide gel was transferred to the Immobilon PVDF membrane using a semi-dry type transfer device. After the transcription, the membrane was blocked and immersed in a primary antibody (anti-Cdc2 rabbit polyclonal antibody: Cell Signaling, anti-Phospho-cdk1 rabbit polyclonal antibody: Sigma-Aldrich) solution at 4 ° C. overnight. Further, after immersing in a secondary antibody (alkaline phosphatase-labeled anti-rabbit IgG antibody: Sigma-Aldrich) solution at room temperature for 2 hours, nitroblue tetrazolium and 5-bromo-4-chloro-indolyl phosphate are used as substrates on the membrane. Cdc2 or phosphorylated Cdc2 (P-Cdc2) was detected. As a result, the compound of the present invention showed inhibition of dephosphorylation of P-Cdc2 (increase in P-Cdc2) at a concentration showing a cell growth inhibitory effect (FIG. 1). From these results, it was suggested that the heterocyclic compound of the present invention inhibits intracellular Plk-1 and exerts a cell growth inhibitory action.

Example 21
Manufacture of tablets The following ingredients were mixed, and the mixture was tableted.

It is a figure which shows the dephosphorylation inhibitory effect of P-Cdc2 in the A549 cell of this invention compound.

Claims (11)

General formula (1)
[Wherein the heterocyclic ring represented by A represents pyridine, pyrimidine, dihydropyrimidine, benzimidazole, benzothiazole, dihydrothiazole or thiazolidine;
The ring represented by B represents benzene or furan;
R ¹ represents a nitro group, an oxo group, a lower alkoxy group, a carboxyl group, a lower alkoxycarbonyl group, a lower alkanoyl group, an amino group, a thioxo group or a cyano group;
R ² represents a halogen atom, a lower alkoxy group, a lower alkoxycarbonyl group, a lower alkanoyl group or a cyano group;
X represents -S-, -O- or -NH-;
l represents an integer of 0 to 3 (however, when l is 2 or more, each R ² may be the same or different);
m represents an integer of 0 or 1;
n represents an integer of 0 to 3 (provided that when n represents 2 or more, each of R ¹ may be the same or different). ;
p shows the integer of 0-3. ]
The polo-like kinase-1 inhibitor which uses the heterocyclic compound represented by these, or its salt as an active ingredient. General formula (2)
R ⁴ represents a phenyl group which may have a substituent, a 1-pyrazolyl group which may have a substituent, or a 4-morpholinyl group which may have a substituent. ]
The polo-like kinase-1 inhibitor which uses the heterocyclic compound represented by these, or its salt as an active ingredient. General formula (3)
[Wherein R ⁵ represents a phenethyl group which may have a substituent or a phenyl group which may have a substituent;
R ⁶ represents a hydrogen atom, R ⁷ represents a hydrogen atom or a cyano group, or R ⁶ and R ⁷ together form a 6-membered cyclic ether optionally substituted by an alkyl group;
R ⁸ represents a hydrogen atom, an amino group or a 4-morpholinyl group. ]
The polo-like kinase-1 inhibitor which uses the heterocyclic compound represented by these, or its salt as an active ingredient. The polo-like kinase-1 inhibitor which uses the heterocyclic compound or its salt chosen from the following as an active ingredient.
N- [2- (4-fluorophenyl) ethyl] -2- (pyrimidin-2-ylsulfanyl) acetamide,
2- (pyridin-2-ylsulfanyl) -N- [2- (2,3,4-trimethoxyphenyl) ethyl] acetamide,
N- [2- (4-fluorophenyl) ethyl] -2- (5-nitropyridin-2-ylsulfanyl) acetamide,
N- [2- (4-fluorophenyl) ethyl] -2- (6-oxo-1,6-dihydropyrimidin-2-ylsulfanyl) acetamide,
N- [2- (4-fluorophenyl) ethyl] -2-[(6-methoxy-1H-benzimidazol-2-ylsulfanyl) acetamide,
N- [2- (4-fluorophenyl) ethyl] -2- (5-nitrobenzothiazol-2-ylsulfanyl) acetamide,
2-[[2- (4-fluorophenyl) ethylcarbamoyl] methylsulfanyl] nicotinic acid,
2-[[2- (4-fluorophenyl) ethylcarbamoyl] methylsulfanyl] nicotinic acid methyl ester,
2- (5-amino-1H-benzimidazol-2-ylsulfanyl) -N- [2- (4-fluorophenyl) ethyl] acetamide hydrochloride,
N- [2- (4-fluorophenyl) ethyl] -2- (2-pyrimidin-2-yloxy) acetamide,
N- [2- (4-fluorophenyl) ethyl] -2- (pyridin-2-yloxy) acetamide,
2- (4,5-dihydrothiazol-2-ylsulfanyl) -N- [2- (4-fluorophenyl) ethyl] acetamide,
N- [2- (4-fluorophenyl) ethyl] -2- (2-pyrimidin-2-ylamino) acetamide,
N- [2- (4-fluorophenyl) ethyl] -2- (2-thioxothiazolidin-3-yl) acetamide,
N- (4-acetylphenyl) -2- (6-amino-3,5-dicyano-2-pyridin-2-ylsulfanyl) acetamide,
2- (6-amino-3,5-dicyanopyridin-2-ylsulfanyl) -N- [2- (4-fluorophenyl) ethyl] acetamide,
2- (6-amino-3,5-dicyanopyridin-2-ylsulfanyl) -N-furan-2-ylmethylacetamide,
2- (6-amino-3,5-dicyanopyridin-2-ylsulfanyl) -N- (4-cyanophenyl) acetamide,
2-amino-6- (2-oxo-2-phenylethylsulfanyl) pyridine-3,5-dicarbonitrile,
1-[[(6-Amino-3,5-dicyano-2-pyridinyl) thio] acetyl] -3,5-dimethyl-1H-pyrazole
2-amino-6- (2-morpholin-4-ylethylsulfanyl) pyridine-3,5-dicarbonitrile,
3,6-diamino-5-cyanothieno [2,3-b] pyridine-2-carboxylic acid [2- (4-fluorophenyl) ethyl] amide,
N- (4-acetylphenyl) -1-amino-8,9-dihydro-8,8-dimethyl-5- (4-morpholinyl) -6H-pyrano [4,3-d] thieno [2,3-b ] Pyridine-2-carboxamide   A prophylactic / therapeutic agent for a polo-like kinase-1 enhancing disease comprising the heterocyclic compound according to any one of claims 1 to 4 or a salt thereof as an active ingredient.   The cancer metastasis inhibitor which uses the heterocyclic compound or its salt of any one of Claims 1-4 as an active ingredient.   The anticancer agent which uses the heterocyclic compound or its salt of any one of Claims 1-4 as an active ingredient.   The therapeutic agent for rheumatoid arthritis which uses the heterocyclic compound or its salt of any one of Claims 1-4 as an active ingredient.   The Alzheimer therapeutic agent which uses the heterocyclic compound or its salt in any one of Claims 1-4 as an active ingredient. A heterocyclic compound selected from the following or a salt thereof.
2- (pyridin-2-ylsulfanyl) -N- [2- (2,3,4-trimethoxyphenyl) ethyl] acetamide,
N- [2- (4-fluorophenyl) ethyl] -2- (5-nitropyridin-2-ylsulfanyl) acetamide,
N- [2- (4-fluorophenyl) ethyl] -2- (6-oxo-1,6-dihydropyrimidin-2-ylsulfanyl) acetamide,
N- [2- (4-fluorophenyl) ethyl] -2-[(6-methoxy-1H-benzimidazol-2-ylsulfanyl) acetamide,
N- [2- (4-fluorophenyl) ethyl] -2- (5-nitrobenzothiazol-2-ylsulfanyl) acetamide,
2-[[2- (4-fluorophenyl) ethylcarbamoyl] methylsulfanyl] nicotinic acid,
2-[[2- (4-fluorophenyl) ethylcarbamoyl] methylsulfanyl] nicotinic acid methyl ester,
2- (5-amino-1H-benzimidazol-2-ylsulfanyl) -N- [2- (4-fluorophenyl) ethyl] acetamide hydrochloride,
N- [2- (4-fluorophenyl) ethyl] -2- (2-pyrimidin-2-yloxy) acetamide,
N- [2- (4-fluorophenyl) ethyl] -2- (pyridin-2-yloxy) acetamide,
2- (4,5-dihydrothiazol-2-ylsulfanyl) -N- [2- (4-fluorophenyl) ethyl] acetamide,
N- [2- (4-fluorophenyl) ethyl] -2- (2-pyrimidin-2-ylamino) acetamide,
N- [2- (4-Fluorophenyl) ethyl] -2- (2-thioxothiazolidin-3-yl) acetamide   The pharmaceutical composition which uses the heterocyclic compound or its salt of Claim 10 as an active ingredient.