JP2008303145A - Cardiotonic comprising grk inhibitor - Google Patents

Abstract

<P>PROBLEM TO BE SOLVED: To provide a new cardiotonic exhibiting an excellent therapeutic action profile on the basis of a new action mechanism, because there is now no "cardiotonic" clearly having safety and a life prognosis-improving action on chronic administration. <P>SOLUTION: This cardiotonic comprises a compound having an action for inhibiting G protein-coupled receptor kinase (GRK) or its prodrug, or its salt (GRK inhibitor). <P>COPYRIGHT: (C)2009,JPO&INPIT

Description

  The present invention relates to a compound having a G protein-coupled receptor kinase (GR protein) inhibitory action, a prodrug thereof, or a salt thereof (hereinafter sometimes abbreviated as GRK inhibitor). The present invention relates to a cardiotonic drug (acute cardiac contractility improving drug).

  Heart failure is a general term for pathological conditions caused by an inability to pump blood that meets the demand for peripheral organ tissue due to abnormal cardiac function. It is one of the cardiovascular diseases with extremely poor prognosis, with a 5-year survival rate of approximately 50% (American Heart Association Statistics 1997), and male patients over 65 years of age with an 8-year survival rate of 20% (American Heart Association) Statistics 2004). In the United States, more than 2% of the total population, with 5 million people suffering from this disease, the number of patients is on the rise. Heart failure treatment has two objectives: improvement of acute symptoms and improvement of chronic life prognosis. Currently, pharmacotherapy is expected to improve acute symptoms, catecholamines, diuretics, digitalis, and PDE inhibition. Drugs such as drugs are expected to have an effect of improving chronic prognosis and survival rate, and angiotensin converting enzyme inhibitors, angiotensin receptor antagonists, β-blockers, aldosterone antagonists, and the like are used. However, the therapeutic effects of these drugs are far from satisfactory, and the only radical treatment that can truly recover from a heart failure state is heart transplantation.

  Drugs that act directly on the heart and improve the cardiac contraction force immediately after drug administration are called `` cardiotonic drugs '', mainly for the aforementioned digitalis, catecholamines, PDE inhibitors, etc. It forms a category as an essential drug for symptom improvement. However, catecholamines and PDE inhibitors are known to accelerate life prognosis in the long run by accelerating the progression of heart failure or increasing the frequency of fatal ventricular arrhythmias. Digitalis is also known to have severe fatal side effects called “digitalis poisoning” and is a placebo-controlled, double-blind, other large-scale clinical trial for 6800 patients with heart failure. Even in the DIG trial, no survival rate improving effect has been observed (Non-patent Document 1). In other words, at present, there are no “cardiotonic drugs” that clearly have the effects of improving the safety and prognosis of chronic administration, and there is a need to create new cardiotonic drugs that exhibit a better medicinal profile based on a new mechanism of action. ing.

  On the other hand, β-receptor desensitization, which is considered to be one of the causes of decreased cardiac function in heart failure in recent years, is GRK2 (G protein-coupled receptor kinase-2, also known as β-receptor kinase-1 ) Receptor phosphorylation caused by long-term suppression of cardiac GRK2 using genetic modification techniques can improve both cardiac function and survival in heart failure model animals Many findings have been reported (Non-Patent Documents 2 and 3). Furthermore, the increase in cardiac GRK2 protein expression has been reported in heart failure model animals and human heart failure patients (Non-patent Document 4). Therefore, inhibition of GRK is different from existing therapeutic agents for heart failure, and heart function is improved and prolonged. It is attracting attention as a new mechanism that can improve prognosis. For example, Harding et al. Crossed a calcequestrin myocardial overexpression heart failure mouse, a model animal of dilated cardiomyopathy, with a mouse that overexpressed βARKct, a GRK inhibitor protein, in the myocardium. It has been reported that inhibition of cardiac GRK has improved cardiac contractility and cardiac relaxation at 7 weeks of age, and prolonged survival has been observed in the long term (Non-Patent Document 5). ). In addition, White et al. And Shaha et al. Reported that βARKct gene transfer into the heart of rabbits with heart failure after myocardial infarction inhibited cardiac GRK and improved cardiac function 1-3 weeks after gene transfer. (Non-Patent Documents 6 and 7).

  However, in studies using these genetic modification techniques, it takes several hours to several days for the introduced gene to express the inhibitory protein and actually exhibit an inhibitory effect, so the observed event is the inhibition of GRK. It is not possible to identify whether it was generated directly by the method or indirectly through another route. In particular, whether acute GRK inhibition can directly improve acute cardiac contractility, that is, whether GRK inhibition is a new “cardiotonic” mechanism, is administered to organs and living bodies. Thus, it was impossible to confirm because there was no low molecular weight compound that could selectively inhibit the action of GRK acutely, and it was completely unknown.

Therefore, although there was a vague expectation that long-term inhibition of GRK would eventually lead to improved cardiac function via multiple pathways, GRK inhibitors acted directly on the heart and became acute immediately after drug administration. Those skilled in the art had no knowledge of having a pharmaceutical use as a “cardiotonic agent” to improve cardiac contraction force.
New England Journal of Medicine, 336, 525-533 Nature Biotechnology, Volume 14, pp. 283-286 Trends in Cardiovascular Medicine, Vol. 9, pp. 77-81 Circulation Research, 74, 206-213 Bulletin of the American Academy of Sciences, Vol. 98, pp. 5809-5814 Bulletin of the American Academy of Sciences, Vol. 97, pp. 5428-5433 Circulation, Vol.103, 1311-1316

  As mentioned above, there are currently no “cardiotonic drugs” that clearly have safety and prognosis-improving effects in chronic administration, and new cardiotonic drugs that have a better drug efficacy profile have been created based on a new mechanism of action. It was sought after.

In the process of diligently searching for drugs that can selectively inhibit GRK by applying to cells, organ specimens, and living organisms, the present inventors have found that a compound having a triazole skeleton has a strong GRK inhibitory action in an enzyme assay. It has been found that it has high selectivity and can inhibit GRK existing in the cell through the cell membrane even when applied from outside the cell. Furthermore, by applying this compound directly to the isolated perfused heart specimen, the cardiac contractile force increases immediately after drug administration, and its action is further increased in the presence of catecholamine, and the heart rate is increased. It was clarified that it hardly occurred. In addition, it was found that by administering the compound intravenously to anesthetized rats, immediately after drug administration, only the systolic force is selectively enhanced with little effect on blood pressure and heart rate, and recovered by discontinuation of administration. It was.
Based on the above results, it was clarified that GRK inhibition acts as a new mechanism of so-called “cardiotonic drugs” that directly affect the heart and improve cardiac contraction force immediately after drug administration, and completed the present invention.

That is, the present invention
(1) a cardiotonic drug containing a GRK inhibitor;
(2) the cardiotonic agent according to (1), wherein the GRK inhibitor is a GRK2 inhibitor;
Etc.

Based on a new mechanism different from known drugs, it was the first time that a GRK inhibitor had a medicinal effect as a “cardiotonic drug” that directly affects the heart and improves cardiac contraction force immediately after drug administration. Based on the above, it was possible to provide “a cardiotonic drug containing a GRK inhibitor”.
In addition, it has been confirmed that GRK inhibitors have the property of hardly causing an increase in heart rate, and it is expected from the action mechanism that they also have a chronic life prognosis improving action.
That is, according to the present invention, an excellent “cardiotonic drug” that has not been available in the past is provided, and a prophylactic / therapeutic agent for heart failure (especially symptomatic heart failure) based on the drug efficacy as a “cardiac drug”. It was demonstrated for the first time that it can actually be applied as a symptom relieving agent for heart failure, a left ventricular function improving agent in heart failure patients, and the like.

The “GRK inhibitor” used in the present invention is not particularly limited as long as it is a compound having a GRK inhibitory action, and is a low molecular compound, a nucleic acid, a protein, a peptide, or a prodrug thereof, or a salt thereof, etc. However, it is advantageous in the practice of the present invention to use one having a strong GRK inhibitory action. For example, when the inhibitory activity of GRK2 is measured by the method disclosed in Test Example 1 described later, the IC ₅₀ value is 2 μM or less, more preferably 0.1 μM or less. However, the present invention is not limited to this.
In addition, the “GRK inhibitor” used in the present invention may have a pharmacological action other than the GRK inhibitory action, but has a high selectivity to the GRK inhibitory action, particularly, the selection to the GRK2 inhibitory action. It is advantageous in the practice of the present invention to use a material having high properties.

  Examples of the GRK inhibitor used in the present invention include compounds represented by the following formula (I) described in International Application No. PCT / JP2005 / 005995, and the like. A GRK inhibitor can be appropriately selected from these to carry out the present invention. The following compounds are merely examples of GRK inhibitors and are not intended to limit the present invention.

〔式中、
環Ａは置換されていてもよい芳香環を示し、
環Ｂは置換されていてもよい５員の含窒素芳香族複素環を示し、
環Ｃは置換されていてもよい含窒素芳香族複素環を示し、
Ｘは置換されていてもよいＣ_1-4アルキレン基を示し、
Ｙは置換されていてもよいイミノ基、−Ｏ−または−Ｓ(Ｏ)ｎ−（ｎは０、１または２を示す。）を示す。〕
で表される化合物もしくはその塩。 Formula (I):

[Where,
Ring A represents an optionally substituted aromatic ring,
Ring B represents an optionally substituted 5-membered nitrogen-containing aromatic heterocycle,
Ring C represents an optionally substituted nitrogen-containing aromatic heterocycle,
X represents an optionally substituted C _1-4 alkylene group,
Y represents an optionally substituted imino group, —O— or —S (O) n— (n represents 0, 1 or 2). ]
Or a salt thereof.

In the formula (I), ring A represents an optionally substituted aromatic ring.
The aromatic ring in the “optionally substituted aromatic ring” represented by ring A is, for example, selected from (i) aromatic cyclic hydrocarbon, (ii) nitrogen atom, sulfur atom and oxygen atom in addition to carbon atom. And aromatic heterocycles preferably containing 1 to 3 heteroatoms.

Examples of the “aromatic cyclic hydrocarbon” include C _6-14 aromatic cyclic hydrocarbons such as benzene and naphthalene.
Examples of the “aromatic heterocycle” include 5- to 6-membered aromatic monocyclic heterocycles such as furan, thiophene, pyridine, pyridazine, pyrimidine, pyrazine, and triazine; A 16-membered (preferably 8- to 12-membered) aromatic condensed heterocyclic ring (preferably 1 to 2 (preferably 1) 5- to 6-membered aromatic monocyclic heterocycle described above is a benzene ring 1. 2 to 3 (preferably 1) heterocycles, or 2 to 3 (preferably 2) the same or different heterocycles of the 5- to 6-membered aromatic monocyclic heterocycle described above Condensed heterocyclic ring); and the like.
Here, as the substituent that the aromatic ring in the “optionally substituted aromatic ring” represented by ring A may have, a halogen atom, an optionally substituted hydrocarbon group, and a substituted An optionally substituted heterocyclic group, an optionally substituted amino group, an optionally substituted imidoyl group, an optionally substituted amidino group, an optionally substituted hydroxy group, and an optionally substituted thiol group Cyano group, nitro group, nitroso group, sulfo group, carbamoyl group which may be substituted, sulfamoyl group which may be substituted, carboxyl group which may be esterified, acyl group, etc. Arbitrary substituents may be substituted by 1 to 5 (preferably 1 to 3) at substitutable positions.

Examples of the “halogen atom” as a substituent that the aromatic ring in the “optionally substituted aromatic ring” represented by ring A may have include fluorine, chlorine, bromine, and iodine.
As the “hydrocarbon group” in the “optionally substituted hydrocarbon group” as the substituent that the aromatic ring in the “optionally substituted aromatic ring” represented by ring A may have, for example, , An aliphatic chain hydrocarbon group, an alicyclic hydrocarbon group (non-aromatic cyclic hydrocarbon group), an aryl group (aromatic hydrocarbon group) and the like.

Examples of the “aliphatic chain hydrocarbon group” as an example of the hydrocarbon group include linear or branched aliphatic hydrocarbon groups such as an alkyl group, an alkenyl group, and an alkynyl group.
Here, examples of the “alkyl group” include C _1-10 alkyl groups (preferably C _1-6 alkyl etc.) such as methyl, ethyl, n-propyl, isopropyl and the like.
Examples of the “alkenyl group” include C _2-6 alkenyl groups such as vinyl, allyl, isopropenyl and the like.
Examples of the “alkynyl group” include C _2-6 alkynyl groups such as ethynyl, 1-propynyl, 2-propynyl and the like.

Examples of the “alicyclic hydrocarbon group” as an example of the hydrocarbon group include saturated or unsaturated alicyclic hydrocarbon groups such as a cycloalkyl group, a cycloalkenyl group, and a cycloalkadienyl group.
Here, examples of the “cycloalkyl group” include C _3-9 cycloalkyl groups such as cyclopropyl and cyclobutyl.
Examples of the “cycloalkenyl group” include C _3-9 cycloalkenyl groups such as 2-cyclopenten-1-yl and 3-cyclopenten-1-yl.
Examples of the “cycloalkadienyl group” include C _4-6 such as 2,4-cyclopentadien-1-yl, 2,4-cyclohexadien-1-yl, 2,5-cyclohexadien-1-yl and the like. And cycloalkadienyl group.

Examples of the “aryl group” as an example of the hydrocarbon group include a monocyclic or condensed polycyclic aromatic hydrocarbon group, and for example, a C _6-14 aryl group such as phenyl, naphthyl, etc. is preferable, among others. A C _6-12 aryl group and the like are particularly preferable.
Moreover, as an example of a hydrocarbon group, the same or different 2 to 3 rings constituting a group selected from the above-described alicyclic hydrocarbon group and aryl group such as 1,2-dihydronaphthyl ( Bicyclic or tricyclic hydrocarbon groups derived from the condensation of preferably two or more rings are also included. Furthermore, bridged hydrocarbon groups such as adamantyl are also included.

The “hydrocarbon group” in the “optionally substituted hydrocarbon group” as the substituent that the aromatic ring in the “optionally substituted aromatic ring” represented by ring A may have Examples of the substituent that may be used include:
(i) a nitro group;
(ii) a hydroxy group, an oxo group;
(iii) a cyano group;
(iv) a carbamoyl group;
(v) a mono- or di-C _1-4 alkyl-carbamoyl group (the alkyl group may be substituted with a halogen atom, a hydroxy group, a C _1-4 alkoxy group, etc.), mono- or di-C _{2 -4} alkenyl-carbamoyl group (this alkenyl group may be substituted with a halogen atom, hydroxy group, C _1-4 alkoxy group, etc.), mono- or di-phenyl-carbamoyl group, mono- or di-aralkyl- Carbamoyl group, C _1-4 alkoxy-carbonyl-carbamoyl group, C _1-4 alkylsulfonyl-carbamoyl group, C _1-4 alkoxy-carbamoyl group, amino-carbamoyl group, mono- or di-C _1-4 alkylamino- A carbamoyl group, a mono- or di-phenylamino-carbamoyl group;
(vi) a carboxyl group;
(vii) a C _1-4 alkoxy-carbonyl group;
(viii) a sulfo group;
(ix) a halogen atom;
(x) optionally halogenated optionally C _1-4 alkoxy groups, optionally substituted by hydroxy C _1-4 alkoxy group, optionally C _1-4 alkoxy group optionally substituted by carboxyl groups, C C _1-4 alkoxy group optionally substituted with _1-4 alkoxy-carbonyl group, C _1-4 alkoxy-C _1-4 alkoxy group, C _1-4 alkoxy-C _1-4 alkoxy-C _1-4 An alkoxy group;
(xi) Phenoxy group, phenoxy-C _1-4 alkyl group, phenoxy-C _1-4 alkoxy group, C _1-4 alkyl-carbonyloxy group, carbamoyloxy group, mono- or di-C _1-4 alkyl-carbamoyl An oxy group;
(xii) selected from a halogen atom, an optionally halogenated C _1-4 alkyl group, optionally C _1-4 alkyl group optionally substituted by also optionally C _1-4 alkoxy and hydroxy halogenated A C _6-12 aryl group which may be substituted with a substituent such as
(xiii) an optionally halogenated phenyl-C _1-4 alkyl group, an optionally halogenated phenyl-C _2-4 alkenyl group, an optionally halogenated phenoxy group, a pyridyloxy group, C _{A 3-10} cycloalkyl-C _1-4 alkoxy group, a C _3-10 cycloalkyl-C _1-4 alkyl group;
(xiv) a C _3-10 cycloalkyl group _optionally substituted with a hydroxyl group, a bicyclic hydrocarbon group in which a C _3-10 cycloalkyl group and a benzene ring are condensed (for example, indanyl), a bridged type A hydrocarbon group (eg, adamantyl, etc.);
(xv) optionally halogenated C _1-4 alkyl group optionally, optionally halogenated C _2-6 also be an alkenyl group, halogenated which may be C _1-4 alkylthio group, substituted with a hydroxy group An optionally substituted C _1-4 alkyl group, a C _1-4 alkylthio group optionally substituted with a hydroxy group;
(xvi) a mercapto group, a thioxo group;
(xvii) a benzyloxy group or a benzylthio group each optionally substituted with a substituent selected from a halogen atom, a carboxyl group, and a C _1-4 alkoxy-carbonyl group;
(xviii) an optionally halogenated phenylthio group, pyridylthio group, phenylthio-C _1-4 alkyl group, pyridylthio-C _1-4 alkyl group;
(xix) an optionally halogenated C _1-4 alkylsulfinyl group;
(xx) an optionally halogenated C _1-4 alkylsulfonyl group, phenylsulfonyl group, phenylsulfonyl -C _1-4 alkyl group;
(xxi) a sulfamoyl group, a mono- or di-C _1-4 alkylsulfamoyl group (the alkyl group may be substituted with a halogen atom, a hydroxy group, a C _1-4 alkoxy group, etc.);
(xxii) amino group, C _1-10 acyl-amino group [C _1-10 acyl may be substituted with a halogen atom, hydroxy group, carboxyl group, etc.], benzyloxycarbonylamino, halogenated A good C _1-6 alkoxy-carbonylamino, carbamoylamino group, mono- or di-C _1-4 alkyl-carbamoylamino group;
(xxiii) mono- or di-C _1-4 alkylamino group (the alkyl group may be substituted with a halogen atom, a hydroxy group, a C _1-4 alkoxy group, etc.), phenylamino, benzylamino;
(xxiv) 4- to 6-membered cyclic amino group, 4- to 6-membered cyclic amino-carbonyl group, 4- to 6-membered cyclic amino-carbonyl-oxy group, 4- to 6-membered cyclic amino-carbonyl-amino group, 4- to 6-membered cyclic group An amino-sulfonyl group, a 4- to 6-membered cyclic amino-C _1-4 alkyl group;
(xxv) a C _1-6 acyl group or a benzoyl group each optionally substituted with a substituent selected from a halogen atom, a carboxyl group, and a C _1-4 alkoxy-carbonyl group;
(xxvi) a benzoyl group optionally substituted by a halogen atom;
(xxvii) a 5- to 10-membered heterocyclic group (the heterocyclic group may be substituted with a C _1-4 alkyl group or the like);
(xxviii) a 5- to 10-membered heterocyclic-carbonyl group (the heterocyclic group may be substituted with a C _1-4 alkyl group or the like);
(xxix) hydroxyimino group, C _1-4 alkoxyimino group;
(xxx) a linear or branched C _1-4 alkylenedioxy group which may be halogenated;
Etc.

  The “hydrocarbon group” may have 1 to 5 (preferably 1 to 3, more preferably 1 or 2) of these substituents at substitutable positions, and has 2 or more. In this case, the substituents may be the same or different.

  As the “heterocyclic group” in the “optionally substituted heterocyclic group” as the substituent that the aromatic ring in the “optionally substituted aromatic ring” represented by ring A may have, for example, In addition, as atoms (ring atoms) constituting the ring system, at least one (preferably 1 to 4) of 1 to 3 (preferably 1 to 2) heteroatoms selected from an oxygen atom, a sulfur atom, a nitrogen atom and the like are used. And more preferably 1 to 2) aromatic heterocyclic groups, saturated or unsaturated non-aromatic heterocyclic groups (aliphatic heterocyclic groups), and the like.

  Examples of the “aromatic heterocyclic group” include 5- to 6-membered aromatic monocyclic heterocyclic groups such as furyl, thienyl, tetrazolyl, pyridyl, pyridazinyl, pyrimidinyl, pyrazinyl, triazinyl and the like; for example, benzofuranyl, indolyl, imidazo An 8- to 16-membered (preferably 8- to 12-membered) aromatic condensed heterocyclic group such as [1,5-a] pyridyl (preferably the 5- to 6-membered aromatic monocyclic heterocyclic group described above is A heterocyclic group condensed with a benzene ring, or a heterocyclic group condensed with the same or different heterocyclic ring of the 5- to 6-membered aromatic monocyclic heterocyclic group described above, and more preferably the 5- to 6-membered heterocyclic group described above And the aromatic monocyclic heterocyclic group is a heterocyclic group condensed with a benzene ring).

  Examples of the “non-aromatic heterocyclic group” include 3 to 8 members (preferably 5 to 6 members) such as pyrrolidinyl, piperidyl, tetrahydropyranyl, morpholinyl, thiomorpholinyl (the sulfur atom may be oxidized), piperazinyl and the like. ) Saturated or unsaturated (preferably saturated) non-aromatic heterocyclic group (aliphatic heterocyclic group), etc .; or 1,2,3,4-tetrahydroquinolyl, 1,2,3,4-tetrahydroiso Examples thereof include non-aromatic heterocyclic groups in which some or all of the double bonds of the aromatic monocyclic heterocyclic group or aromatic condensed heterocyclic group described above are saturated, such as quinolyl.

  The “heterocyclic group” in the “optionally substituted heterocyclic group” as the substituent that the aromatic ring in the “optionally substituted aromatic ring” represented by ring A may have Examples of the substituent that may be present include, for example, the “hydrocarbon group” in the “optionally substituted hydrocarbon group” as the substituent of the “optionally substituted aromatic ring” represented by ring A. And the same number of the same groups as the substituents which may be present.

The “optionally substituted amino group” and the “optionally substituted imidoyl group” as the substituents that the aromatic ring may have in the “optionally substituted aromatic ring” represented by ring A Amino group, imidoyl group, amidino group, hydroxy group and thiol group in "optionally substituted amidino group", "optionally substituted hydroxy group" and "optionally substituted thiol group" Examples of the substituent that may have, for example,
(i) a halogen atom, a C _1-6 alkylthio which may be halogenated, phenyl (said phenyl, halogen atom, optionally halogenated C _1-6 alkyl, optionally C ₁ may be halogenated _A lower alkyl group which may be substituted with a substituent selected from naphthyl which may be halogenated and C _3-10 cycloalkyl, etc.
(ii) an optionally halogenated phenyl group;
(iii) an optionally halogenated C _3-10 cycloalkyl group;
(iv) Acyl group [optionally halogenated C _1-6 alkanoyl, phenylacetyl (the phenyl may be substituted with a halogen atom, optionally halogenated C _1-6 alkyl, etc.) An optionally halogenated benzoyl, an optionally halogenated C _1-6 alkylsulfonyl group, an optionally halogenated benzenesulfonyl, toluenesulfonyl, etc.];
(v) optionally halogenated C _1-6 alkoxy optionally - carbonyl group, optionally substituted by phenyl C _1-6 alkoxy - carbonyl group;
(vi) mono- or di-aralkyl-carbamoyl group, mono- or di-C _1-6 alkyl-carbamoyl group, phenyl-carbamoyl group;
(vii) a heterocyclic group (for example, the same as the “heterocyclic group” in the above-mentioned “optionally substituted heterocyclic group”);
Etc. The “amino group” in the “optionally substituted amino group” as the substituent is an imidoyl group which may be substituted (for example, C _1-6 alkylimidoyl, C _1-6 alkoxyimidoyl, C _{1 -6} alkylthioimidoyl, amidino, etc.), an amino group optionally substituted with 1 to 2 C _1-6 alkyls, and the like.
In the case of “optionally substituted amino group”, “optionally substituted imidoyl group” and “optionally substituted amidino group”, two substituents are combined with the nitrogen atom. In some cases, a “cyclic amino group” may be formed. Examples of the cyclic amino group include 1-azetidinyl, 1-pyrrolidinyl, piperidino, thiomorpholino, morpholino, 1-piperazinyl, and lower alkyl at the 4-position. And a cyclic amino group having 3 to 8 members (preferably 5 to 6 members) such as 1-piperazinyl, 1-pyrrolyl and 1-imidazolyl, which may have a group, an aralkyl group, an aryl group, and the like.
In the “optionally substituted aromatic ring” represented by ring A, two adjacent substituents present on the aromatic ring are combined to form a linear or branched C _1-4 alkylenedioxy A group or the like may be formed.

As the “optionally substituted carbamoyl group” as the substituent that the aromatic ring in the “optionally substituted aromatic ring” represented by ring A may have, in addition to an unsubstituted carbamoyl group, N-monosubstituted carbamoyl group and N, N-disubstituted carbamoyl group can be mentioned.
Examples of the substituent of the “N-monosubstituted carbamoyl group” include an optionally substituted hydrocarbon group, an optionally substituted heterocyclic group, and an optionally substituted amino.
Examples of the “optionally substituted hydrocarbon group” as an example of the substituent of the “N-monosubstituted carbamoyl group” include “the optionally substituted aromatic ring” represented by ring A as “ Examples thereof include the same groups as the “optionally substituted hydrocarbon group”.
Examples of the “optionally substituted heterocyclic group” as an example of the substituent of the “N-monosubstituted carbamoyl group” include “the optionally substituted aromatic ring” represented by ring A as “ Examples thereof include the same groups as the “optionally substituted heterocyclic group”.
Examples of the “optionally substituted amino group” as an example of the substituent of the “N-monosubstituted carbamoyl group” include “substitution” as a substituent in the “optionally substituted aromatic ring” represented by ring A Examples thereof include the same groups as “optionally substituted amino group”.
The “N, N-disubstituted carbamoyl group” means a carbamoyl group having two substituents on the nitrogen atom, and one example of the substituent is the above-mentioned “N-monosubstituted carbamoyl group”. Examples of the substituent are the same, and examples of the other include a lower alkyl group, a C _3-6 cycloalkyl group, a C _7-10 aralkyl group, and the like. In addition, two substituents may form a cyclic amino group together with a nitrogen atom. In such a case, examples of the cyclic aminocarbamoyl group include 1-azetidinylcarbonyl, 1-pyrrolidinyl. Rucarbonyl, piperidinocarbonyl, morpholinocarbonyl, thiomorpholinocarbonyl (sulfur atom may be oxidized), 1-piperazinylcarbonyl, and lower alkyl group, aralkyl group, aryl group, etc. at the 4-position And a 3- to 8-membered (preferably 5 to 6-membered) cyclic amino-carbonyl group such as 1-piperazinylcarbonyl may be mentioned.

As the “optionally substituted sulfamoyl group” as the substituent that the aromatic ring in the “optionally substituted aromatic ring” represented by ring A may have, in addition to an unsubstituted sulfamoyl group, Examples thereof include N-monosubstituted sulfamoyl groups and N, N-disubstituted sulfamoyl groups.
Examples of the substituent of the “N-monosubstituted sulfamoyl group” include the same groups as the substituent of the “N-monosubstituted carbamoyl group”.
The “N, N-disubstituted sulfamoyl group” means a sulfamoyl group having two substituents on the nitrogen atom, and one example of the substituent is the above-mentioned “N-monosubstituted sulfamoyl group”. Examples of the substituent are the same, and examples of the other include a lower alkyl group, a C _3-6 cycloalkyl group, a C _7-10 aralkyl group, and the like. In addition, two substituents may form a cyclic amino group together with a nitrogen atom. In such a case, examples of the cyclic aminosulfamoyl group include 1-azetidinylsulfonyl, 1- Pyrrolidinylsulfonyl, piperidinosulfonyl, morpholinosulfonyl, thiomorpholinosulfonyl (sulfur atom may be oxidized), 1-piperazinylsulfonyl, and lower alkyl group, aralkyl group, aryl group and the like at the 4-position Examples thereof include a 3- to 8-membered (preferably 5 to 6-membered) cyclic amino-sulfonyl group such as 1-piperazinylsulfonyl.

As the “optionally esterified carboxyl group” as the substituent that the aromatic ring in the “optionally substituted aromatic ring” represented by ring A may have, in addition to a free carboxyl group, For example, a lower alkoxycarbonyl group, an aryloxycarbonyl group, an aralkyloxycarbonyl group and the like can be mentioned.
Examples of the “lower alkoxycarbonyl group” include C _1-6 alkoxy-carbonyl such as methoxycarbonyl, ethoxycarbonyl, propoxycarbonyl, etc., among which C _1-3 alkoxy-carbonyl and the like are preferable.
As the “aryloxycarbonyl group”, for example, C _6-12 aryl-oxycarbonyl such as phenoxycarbonyl, 1-naphthoxycarbonyl, 2-naphthoxycarbonyl and the like are preferable.
As the “aralkyloxycarbonyl group”, for example, C _7-10 aralkyl-oxycarbonyl such as benzyloxycarbonyl, phenethyloxycarbonyl and the like (preferably C _6-10 aryl-C _1-4 alkoxy-carbonyl etc.) are preferable. .
The “lower alkoxycarbonyl group”, “aryloxycarbonyl group”, and “aralkyloxycarbonyl group” may have a substituent, and the substituent may be “substituted” represented by ring A. The same number of the same groups as those mentioned as the substituents that the “hydrocarbon group” in the “optionally substituted hydrocarbon group” as the substituent in the “aromatic ring” may have is used.

As the “acyl group” as the substituent that the aromatic ring in the “optionally substituted aromatic ring” represented by ring A may have, an acyl group derived from carboxylic acid, an acyl group derived from sulfinic acid, Examples include acyl groups derived from sulfonic acids and acyl groups derived from phosphonic acids.
The “acyl group derived from carboxylic acid” includes a hydrogen atom, an optionally substituted hydrocarbon group (for example, “substituted” in the “optionally substituted aromatic ring” represented by ring A). The same as the “optionally substituted hydrocarbon group” or a heterocyclic group which may be substituted (for example, “substituted” in the “optionally substituted aromatic ring” represented by ring A) A group similar to “optionally substituted heterocyclic group” and a carbonyl (—C (O) —), such as formyl; acetyl, propionyl, butyryl, isobutyryl, valeryl, isovaleryl, pivaloyl, Chains that may be halogenated, such as hexanoyl, cyclobutanecarbonyl, cyclopentanecarbonyl, cyclohexanecarbonyl, crotonyl, trifluoroacetyl, etc. Or cyclic C _2-8 alkanoyl; benzoyl, nicotinoyl, isonicotinoyl and the like. Among them, C _2-5 alkanoyl such as acetyl, propionyl, butyryl, valeryl, pivaloyl and the like are preferable.
As the “acyl group derived from sulfinic acid”, an optionally substituted hydrocarbon group (for example, “substituted” in the “optionally substituted aromatic ring” represented by ring A may be A group similar to “good hydrocarbon group” or the like, or a heterocyclic group which may be substituted (for example, “substituted” in the “optionally substituted aromatic ring” represented by ring A) A group similar to “a good heterocyclic group” and a sulfinyl group (—S (O) —), for example, methanesulfinyl, ethanesulfinyl, propanesulfinyl, cyclopropanesulfinyl, cyclopentanesulfinyl, cyclohexanesulfinyl. C _1-6 alkylsulfinyl halides which may be chain or cyclic and the like; benzene sulfinyl, toluene sulfinyl Etc. The.
The “acyl group derived from sulfonic acid” is an optionally substituted hydrocarbon group (for example, “substituted” in the “optionally substituted aromatic ring” represented by ring A). A group similar to “good hydrocarbon group” or the like, or a heterocyclic group which may be substituted (for example, “substituted” in the “optionally substituted aromatic ring” represented by ring A) A group similar to “a good heterocyclic group” and a sulfonyl group (—S (O) ₂ —), such as methanesulfonyl, ethanesulfonyl, propanesulfonyl, cyclopropanesulfonyl, cyclopentanesulfonyl, cyclohexane Examples thereof include linear or cyclic C _1-6 alkylsulfonyl which may be halogenated such as sulfonyl; benzenesulfonyl, toluenesulfonyl and the like.
Examples of the “acyl group derived from phosphonic acid” include dimethylphosphono, diethylphosphono, diisopropylphosphono, dibutylphosphono, 2-oxide-1,3,2-dioxaphosphinan-2-yl and the like. And mono- or di-C _1-4 alkylphosphono which may form a ring.

As the substituent that the aromatic ring in ring A may have,
A halogen atom;
A C _1-6 alkyl group (the C _1-6 alkyl group is a halogen atom, hydroxy, carbamoyl, mono- or di-benzylcarbamoyl, mono- or di-phenethylcarbamoyl, carboxyl, C _1-4 alkoxy-carbonyl, halogen C _1-4 alkoxy, amino, mono- or di-C _1-4 alkylamino, C _1-6 alkanoylamino, optionally halogenated phenyl, optionally halogenated benzoyl Etc.);
A C _1-6 alkenyl group (the C _1-6 alkenyl group may be substituted with an optionally halogenated benzoyl or the like);
Phenyl group (the phenyl is a halogen atom, hydroxy, carbamoyl, carboxyl, sulfo group, C _1-4 alkoxy-carbonyl, optionally halogenated C _1-4 alkoxy, optionally halogenated C _1- Optionally substituted with ₄ alkyl, amino, mono- or di-C _1-4 alkylamino, C _1-6 alkanoylamino, etc.);
A non-aromatic heterocyclic group (preferably oxazoline, the non-aromatic heterocyclic group may be substituted with an oxo group or the like);
An amino group [the amino group is an optionally halogenated C _1-6 alkyl, an optionally halogenated C _1-6 alkanoyl, an optionally halogenated benzoyl, phenylacetyl (the phenyl is Substituted with a halogen atom, optionally halogenated C _1-6 alkyl, etc.), mono- or di-benzylcarbamoyl group, mono- or di-phenethylcarbamoyl group, substituted with phenyl A good C _1-6 alkoxy-carbonyl group, optionally substituted by a halogenated C _1-6 alkylsulfonyl, etc.];
A hydroxy group [the hydroxy group is selected from phenyl (the phenyl may be substituted with a halogen atom, optionally halogenated C _1-6 alkyl, etc.) and an optionally halogenated naphthyl; A C _1-6 alkyl which may be substituted with a substituent, a phenyl which may be halogenated, etc.]
A C _1-4 alkylenedioxy group;
A thiol group (the thiol group may be substituted with a halogen atom or optionally halogenated C _1-6 alkyl, optionally halogenated phenyl, etc.);
A carbamoyl group;
An N-monosubstituted carbamoyl group [eg,
N—C _1-6 alkyl (wherein the alkyl may be substituted with a halogen atom, hydroxy or the like) -carbamoyl group;
N-phenyl (the phenyl may be substituted with a halogen atom, C _1-4 alkyl which may be halogenated, etc.)-Carbamoyl group;
An N-nonaromatic heterocycle (preferably tetrahydropyranyl) -carbamoyl group;
N-bi- or tricyclic hydrocarbon (preferably tetrahydronaphthyl, indanyl, fluorenyl, the bi- or tricyclic hydrocarbon may be substituted with hydroxy or the like) -carbamoyl group;
N—C _6-12 aryl (wherein the aryl is a halogen atom, C _1-4 alkyl which may be halogenated, C _1-4 alkoxy which may be halogenated, C which may be substituted with hydroxy) _1-4 may be substituted with _1-4 alkyl or the like) -C _1-6 alkyl (the C _1-6 alkyl may be substituted with hydroxy, carbamoyl or the like) -carbamoyl group;
An N-5 to 10-membered heterocyclic ring (preferably pyridyl, furyl, thienyl, indolyl, the heterocyclic ring may be substituted with C _1-4 alkyl, etc.)-C _1-6 alkyl-carbamoyl group;
N—C _3-10 cycloalkyl (wherein the C _3-10 cycloalkyl may be substituted with hydroxy or the like) —C _1-6 alkyl-carbamoyl group;
N-bicyclic hydrocarbon-C _1-6 alkyl-carbamoyl;
An N-adamantyl-C _1-6 alkyl-carbamoyl group;
An N-phenoxy-C _1-6 alkyl-carbamoyl group;
N-phenylamino-C _1-6 alkyl-carbamoyl group and the like];
N- disubstituted carbamoyl group [for example, N-C _1-6 alkyl -N- phenyl -C _1-6 alkyl - carbamoyl group;
5- to 6-membered cyclic amino (preferably forming a piperidine ring) -carbonyl group (preferably 1,2,3,4-tetrahydroisoquinolin-2-ylcarbonyl) and the like];
A sulfamoyl group;
N-monosubstituted sulfamoyl group [for example, N-phenyl (the phenyl may be substituted with a halogen atom, C _1-4 alkyl which may be halogenated, etc.)-C _1-6 alkyl-sulfamoyl] ;
Carboxyl group;
A C _1-6 alkoxy-carbonyl group;
An optionally halogenated C _2-8 alkanoyl group;
An optionally halogenated C _1-6 alkylsulfinyl group;
A benzylsulfinyl group;
An optionally halogenated C _1-6 alkylsulfonyl group;
A benzylsulfonyl group;
Etc. are preferred.

  Ring A is preferably an optionally substituted benzene ring or an optionally substituted pyridine ring. Among them, an optionally substituted benzene ring is more preferable, and an optionally substituted carbamoyl group is 3 A benzene ring which may be substituted at the position is particularly preferred. In particular, a benzene ring which may be substituted at the 3-position with an N-monosubstituted carbamoyl group is most preferable.

In the formula (I), X represents an optionally substituted C _1-4 alkylene group.
The C _1-4 alkylene group in the "optionally substituted C _1-4 alkylene group" represented by X, methylene, ethylene, trimethylene, tetramethylene and the like.
Examples of the substituent that the C _1-4 alkylene group may have include “substitution” as the substituent that the aromatic ring in the “optionally substituted aromatic ring” represented by ring A may have. The same groups as the substituents that the hydrocarbon group in the “optionally hydrocarbon group” may have are mentioned (however, it is preferably not an oxo group), and these substituents can be substituted. 1 to 3 (preferably 1) may be substituted at any position. As the substituent that the C _1-4 alkylene group may have,
A halogen atom;
A hydroxy group;
A carbamoyl group;
Carboxyl group;
A C _1-4 alkoxy-carbonyl group;
An optionally halogenated C _1-4 alkoxy group;
An optionally halogenated C _1-4 alkyl group;
An optionally halogenated phenyl group;
An amino group;
Mono- or di-C _1-4 alkylamino groups;
Etc. are preferred.
X is preferably an optionally substituted methylene, particularly preferably unsubstituted methylene.

In formula (I), ring B represents an optionally substituted 5-membered nitrogen-containing aromatic heterocycle.
Examples of the “5-membered nitrogen-containing aromatic heterocycle” include pyrrole, oxazole, isoxazole, thiazole, isothiazole, imidazole, piperazole, oxadiazole (1,2,3-oxadiazole, 1,2 , 4-oxadiazole, 1,3,4-oxadiazole), furazane, oxadiazole (1,2,3-thiadiazole, 1,2,4-thiadiazole, 1,3,4-thiadiazole), triazole (1,2,3-triazole, 1,2,4-triazole), tetrazole and the like. Preferred are pyrrole, pyrazole, imidazole, triazole, thiazole, isoxazole, oxadiazole and thiadiazole.

Examples of the substituent that the “5-membered nitrogen-containing aromatic heterocycle” may have include the same groups as the substituents in the “optionally substituted aromatic ring” represented by the ring A described above. 1 to 3 (preferably 1) of these substituents may be substituted at any substitutable position. As the substituent that the “5-membered nitrogen-containing aromatic heterocycle” may have,
A halogen atom;
C _1-6 alkyl group (the C _1-6 alkyl group is a halogen atom, hydroxy, carbamoyl, carboxyl, halogenated phenyl, C _1-4 alkoxy-carbonyl, sulfamoyl, sulfo group, halogenated C _1-4 alkoxy, benzyloxy, amino, mono- or di-C _1-4 alkylamino, C _1-6 alkanoylamino, optionally halogenated C _1-4 alkylthio, halogenated unprotected C _1-4 alkylsulfinyl, it may also be substituted by also good C _1-4 alkylsulfonyl such as halogenated);
Optionally halogenated phenyl;
Etc. are preferred.

で表される基が、

（式中、Ｒ^１は水素原子、置換されていてもよい炭化水素基、置換されていてもよい複素環基またはアシル基を示し、Ｒ^２およびＲ^３はそれぞれ独立して水素原子、ハロゲン原子（例、フッ素、塩素、臭素、ヨウ素）、置換されていてもよい炭化水素基、置換されていてもよい複素環基、置換されていてもよいアミノ基、置換されていてもよいヒドロキシ基、ニトロ基、シアノ基、エステル化されていてもよいカルボキシル基、置換されていてもよいカルバモイル基またはアシル基を示し、Ｘは前記と同意義を示す）であることが好ましい。 Ring B may be bonded to rings C and X at any substitutable position of the “5-membered nitrogen-containing aromatic heterocycle”, and among them, the “5-membered nitrogen-containing aromatic heterocycle” Rings C and X are preferably bonded to the constituent carbon atoms.
Among these, in formula (I), the formula:

The group represented by

(Wherein R ¹ represents a hydrogen atom, an optionally substituted hydrocarbon group, an optionally substituted heterocyclic group or an acyl group; R ² and R ³ each independently represent a hydrogen atom, a halogen atom; (Eg, fluorine, chlorine, bromine, iodine), an optionally substituted hydrocarbon group, an optionally substituted heterocyclic group, an optionally substituted amino group, an optionally substituted hydroxy group, It is preferably a nitro group, a cyano group, an optionally esterified carboxyl group, an optionally substituted carbamoyl group or an acyl group, and X is as defined above.

As the “optionally substituted hydrocarbon group” represented by R ¹ , “substitution” as the substituent that the aromatic ring in the “optionally substituted aromatic ring” represented by ring A may have Examples thereof include the same groups as “optionally substituted hydrocarbon group”.
The “optionally substituted heterocyclic group” represented by R ¹ is “substituted” as the substituent that the aromatic ring in the “optionally substituted aromatic ring” represented by ring A may have. Examples thereof include the same groups as “optionally substituted heterocyclic group”.
Examples of the “acyl group” represented by R ¹ include the same groups as the “acyl group” as a substituent that the aromatic ring in the “optionally substituted aromatic ring” represented by ring A may have, etc. Is mentioned.

As R ¹ ,
Hydrogen atom;
C _1-6 alkyl group (the C _1-6 alkyl group is a halogen atom, hydroxy, carbamoyl, carboxyl, halogenated phenyl, C _1-4 alkoxy-carbonyl, sulfamoyl, sulfo group, halogenated C _1-4 alkoxy, benzyloxy, amino, mono- or di-C _1-4 alkylamino, C _1-6 alkanoylamino, optionally halogenated C _1-4 alkylthio, halogenated unprotected C _1-4 alkylsulfinyl, it may also be substituted by also good C _1-4 alkylsulfonyl such as halogenated);
Optionally halogenated phenyl;
Etc. are preferred.

As the “optionally substituted hydrocarbon group” represented by R ² and R ³ , the aromatic ring in the “optionally substituted aromatic ring” represented by ring A may be And the same groups as the “optionally substituted hydrocarbon group”.
As the “optionally substituted heterocyclic group” for R ² and R ³ , the aromatic ring in the “optionally substituted aromatic ring” for ring A may have And the same groups as the “optionally substituted heterocyclic group”.
As the “optionally substituted amino group” represented by R ² and R ³ , the aromatic ring in the “optionally substituted aromatic ring” represented by ring A may be Examples thereof include a group similar to the “optionally substituted amino group”.
As the “optionally substituted hydroxy group” represented by R ² and R ³ , the aromatic ring in the “optionally substituted aromatic ring” represented by ring A may be Examples thereof include the same groups as the “optionally substituted hydroxy group”.
As the “optionally esterified carboxyl group” represented by R ² and R ³ , as the substituent that the aromatic ring in the “optionally substituted aromatic ring” represented by ring A may have And the same groups as those of “optionally esterified carboxyl group”.
As the “optionally substituted carbamoyl group” represented by R ² and R ³ , the aromatic ring in the “optionally substituted aromatic ring” represented by ring A may be Examples thereof include the same groups as the “optionally substituted carbamoyl group”.
The “acyl group” represented by R ² and R ³ is the same as the “acyl group” as the substituent that the aromatic ring in the “optionally substituted aromatic ring” represented by ring A may have. And the like.

R ² and R ³ are each preferably a hydrogen atom, a halogen atom, an optionally substituted hydrocarbon group, etc.
Hydrogen atom;
A halogen atom;
C _1-6 alkyl group (the C _1-6 alkyl group is a halogen atom, hydroxy, carbamoyl, carboxyl, halogenated phenyl, C _1-4 alkoxy-carbonyl, sulfamoyl, sulfo group, halogenated C _1-4 alkoxy, benzyloxy, amino, mono- or di-C _1-4 alkylamino, C _1-6 alkanoylamino, optionally halogenated C _1-4 alkylthio, halogenated unprotected C _1-4 alkylsulfinyl, it may also be substituted by also good C _1-4 alkylsulfonyl such as halogenated);
Optionally halogenated phenyl;
Etc. are particularly preferred.

In formula (I), Y represents an optionally substituted imino group, —O— or —S (O) n— (n represents 0, 1 or 2).
As the “optionally substituted imino group” represented by Y, the formula: —N (R ⁴ ) — (wherein R ⁴ is a hydrogen atom, an optionally substituted hydrocarbon group, a substituted A divalent group represented by a heterocyclic group which may be substituted, a carboxyl group which may be esterified, a carbamoyl group which may be substituted or an acyl group).
As the “optionally substituted hydrocarbon group” represented by R ⁴ , “substitution” as the substituent that the aromatic ring in the “optionally substituted aromatic ring” represented by ring A may have Examples thereof include the same groups as “optionally substituted hydrocarbon group”.
The “optionally substituted heterocyclic group” represented by R ⁴ is the “substituted” as the substituent that the aromatic ring in the “optionally substituted aromatic ring” represented by ring A may have. Examples thereof include the same groups as “optionally substituted heterocyclic group”.
As the “optionally esterified carboxyl group” represented by R ⁴ , the “ester” as the substituent that the aromatic ring in the “optionally substituted aromatic ring” represented by ring A may have Examples thereof include the same groups as “optionally converted carboxyl group”.
As the “optionally substituted carbamoyl group” represented by R ⁴ , “the substituted carbamoyl group” as the substituent which the aromatic ring in the “optionally substituted aromatic ring” represented by ring A may have Examples thereof include the same groups as the “optional carbamoyl group”.
Examples of the “acyl group” represented by R ⁴ include the same groups as the “acyl group” as the substituent that the aromatic ring in the “optionally substituted aromatic ring” represented by ring A may have, etc. Is mentioned.

Y is preferably an optionally substituted imino group (for example, a group represented by the formula: —N (R ⁴ ) —), among which a C _1-4 alkyl group or a C _1-4 alkoxy group is substituted. In an imino group (that is, —N (R ⁴ ) — which may be substituted with an optionally substituted phenyl group, carbamoyl group, optionally halogenated C _2-8 alkanoyl group, benzoyl group, R ⁴ is a hydrogen atom, a C _1-4 alkyl group, a phenyl group optionally substituted with a C _1-4 alkoxy group, a carbamoyl group, an optionally halogenated C _2-8 alkanoyl group, a benzoyl group, etc.) Are more preferable, and R ⁴ is particularly preferably a hydrogen atom or a C _1-4 alkyl group.
When Y is a substituted imino group, the substituent of the imino group and the substituent of the “aromatic ring” in ring A may be combined to form a ring (for example, tetrahydroquinoline).

In formula (I), ring C represents an optionally substituted nitrogen-containing aromatic heterocycle.
Examples of the “nitrogen-containing aromatic heterocycle” in the “optionally substituted nitrogen-containing aromatic heterocycle” represented by ring C include pyrrole, oxazole, isoxazole, thiazole, isothiazole, imidazole, pyrazole, 1 , 2,4-thiadiazole, 1,3,4-thiadiazole), triazole (1,2,3-triazole, 1,2,4-triazole), tetrazole, pyridine, pyridazine, pyrimidine, pyrazine, triazine (1,2 , 4-triazine, 1,3,5-triazine) ring and the like 5- or 6-membered nitrogen-containing aromatic monocyclic heterocycle; for example, indole, isoindole, pyrazolo [1,5-a] pyridine, imidazo [ 1,2-a] pyridine, imidazo [1,5-a] pyridine, imidazo [1,2-b] pyridazine, imidazo [1,2-a] pyrimidine 8- to 16-membered (preferably 8- to 12-membered) nitrogen-containing aromatics such as 1,2,4-triazolo [4,3-a] pyridine and 1,2,4-triazolo [4,3-b] pyridazine Examples thereof include fused heterocycles, and 5- to 6-membered nitrogen-containing aromatic monocyclic heterocycles are preferable.

Examples of the substituent that the “nitrogen-containing aromatic heterocycle” may have include the same groups as the substituents in the “optionally substituted aromatic ring” represented by the ring A described above, and the like. 1 to 4 (preferably 1 to 2) of these substituents may be substituted at any substitutable position. As the substituent that the “nitrogen-containing aromatic heterocycle” may have,
A halogen atom;
A hydroxy group;
A carbamoyl group;
Carboxyl group;
A C _1-6 alkoxy-carbonyl group sulfamoyl group;
A sulfo group;
An optionally halogenated C _1-6 alkoxy group;
A C _1-6 alkyl group (the C _1-6 alkyl group may be substituted with a halogen atom, a hydroxy group, a carbamoyloxy group, etc.);
An optionally halogenated phenyl group;
An amino group;
Mono- or di-C _1-6 alkylamino group [the alkyl group is a halogen atom, optionally halogenated C _1-6 alkoxy, optionally halogenated C _1-6 alkylthio, phenyl (including Phenyl is selected from a halogen atom, C _1-6 alkyl which may be halogenated, C _1-6 alkoxy which may be halogenated, etc.) and C _3-10 cycloalkyl Optionally substituted with a substituent];
A mono- or di-phenylamino group (the phenyl may be halogenated);
A mono- or di-C _3-10 cycloalkylamino group;
An optionally halogenated C _1-6 alkanoylamino group;
An optionally halogenated benzoylamino group;
A phenylacetylamino group;
A mono- or di-C _1-6 alkyl-carbamoylamino group;
An optionally halogenated phenyl-carbamoylamino group;
An optionally halogenated phenyl-carbamoyl group;
An aromatic heterocycle (preferably thiophene) -carbamoyl group;
An optionally halogenated C _1-6 alkylthio group;
An optionally halogenated C _1-6 alkylsulfinyl group;
An optionally halogenated C _1-6 alkylsulfonyl group;
Etc. are preferred.

In addition, when the “nitrogen-containing aromatic heterocycle” in the “optionally substituted nitrogen-containing aromatic heterocycle” represented by ring C has two or more substituents adjacent to each other, the substituents are bonded to each other. A ring (for example, C _4-8 cycloalkane ring such as cyclopentane, cyclohexane, cycloheptane, benzene ring, etc.) may be formed.
Ring C may be bonded to ring B at any substitutable position of the “nitrogen-containing aromatic heterocycle”, and among them, the carbon atom constituting the “nitrogen-containing aromatic heterocycle” and ring B Bonding is preferred.

で表される基が、

〔式中、環Ｃ’および環Ｃ”はそれぞれ置換されていてもよい。〕であることが好ましい。
環Ｃ’および環Ｃ”がそれぞれ有していてもよい置換基としては、環Ｃが有していてもよい置換基と同様の基が挙げられる。
ピリジン環を構成する窒素原子は、酸化されていてもよい。 Ring C is preferably an optionally substituted pyridine ring or an optionally substituted pyrimidine ring. Among them, in formula (I), the formula:

The group represented by

[Wherein, ring C ′ and ring C ″ may each be substituted.]
Examples of the substituent that each of ring C ′ and ring C ″ may have include the same groups as the substituent that ring C may have.
The nitrogen atom constituting the pyridine ring may be oxidized.

Specific examples of the compound represented by the formula (I) include the following.
N-benzyl-3-[[[5- (4-pyridyl) -1H-1,2,4-triazol-3-yl] methyl] amino] benzamide;
4-chloro-3-phenylacetamido-N-[[5- (4-pyridyl) -1H-1,2,4-triazol-3-yl] methyl] aniline,
N-benzyl-3-[[[5- (4-pyridyl) isoxazol-3-yl] methyl] amino] benzamide,
N-benzyl-N ′-[3-[[[5- (4-pyridyl) -1H-1,2,4-triazol-3-yl] methyl] amino] phenyl] urea,
4-chloro-N-[[5- (4-pyridyl) pyrazol-3-yl] methyl] aniline,
N-benzyl-3-[[[5- (4-pyridyl) pyrazol-3-yl] methyl] amino] benzamide,
3- (methylthio) -N-[[5- (4-pyridyl) -1H-1,2,4-triazol-3-yl] methyl] aniline,
N- (3-methoxybenzyl) -3-[[[5- (4-pyridyl) -1H-1,2,4-triazol-3-yl] methyl] amino] benzamide,
3-[[[4-Methyl-5- (4-pyridyl) -4H-1,2,4-triazol-3-yl] methyl] amino] -N- [2- (trifluoromethyl) benzyl] benzamide,
N-benzyl-3-[[[4-methyl-5- (4-pyridyl) pyrazol-3-yl] methyl] amino] benzamide,
N- (1-Indanyl) -3-[[[4- (2,2,2-trifluoroethyl) -3- (4-pyridyl) -4H-1,2,4-triazol-5-yl] methyl ] Amino] benzamide,
N- [2,6-difluorobenzyl] -3-[[[4-methyl-3- (4-pyrimidyl) -4H-1,2,4-triazol-5-yl] methyl] amino] benzamide;
N- (2,6-difluorobenzyl) -3-[[[4-propyl-3- (4-pyrimidyl) -4H-1,2,4-triazol-5-yl] methyl] amino] benzamide;
N-[(R) -1-phenylethyl] -3-[[[5- (4-pyridyl) thiazol-2-yl] methyl] amino] benzamide, and N-[(R) -1-phenylethyl] -3-[[[[5- (4-pyridyl) -1,3,4-thiadiazol-2-yl] methyl] amino] benzamide

  Examples of the salt of compound (I) include metal salts, ammonium salts, salts with organic bases, salts with inorganic acids, salts with organic acids, salts with basic or acidic amino acids, and the like. Of these, pharmaceutically acceptable salts are preferred. For example, when the compound has an acidic functional group, an inorganic salt such as an alkali metal salt (eg, sodium salt, potassium salt), an alkaline earth metal salt (eg, calcium salt, magnesium salt, barium salt), When the compound has a basic functional group, for example, a salt with an inorganic acid such as hydrochloric acid, hydrobromic acid, nitric acid, sulfuric acid, phosphoric acid, or acetic acid, phthalic acid, fumaric acid And salts with organic acids such as oxalic acid, tartaric acid, maleic acid, citric acid, succinic acid, methanesulfonic acid and p-toluenesulfonic acid.

  The above-mentioned compound (I) can be produced by appropriately adapting, for example, the method disclosed in Reference Example 1 described later and other methods usually used in organic synthesis.

  Compound (I) may be used as a prodrug. A prodrug of compound (I) is a compound that is converted to compound (I) by a reaction with an enzyme, gastric acid, or the like under physiological conditions in vivo, that is, compound (I) that is enzymatically oxidized, reduced, hydrolyzed, etc. A compound that changes to compound (I) by hydrolysis or the like due to gastric acid or the like.

When a GRK inhibitor is used as a cardiotonic drug based on the present invention, a pharmacologically acceptable carrier is mixed with the GRK inhibitor alone or according to a conventional method (for example, the method described in the Japanese Pharmacopoeia). Pharmaceutical compositions such as tablets (including sugar-coated tablets and film-coated tablets), powders, granules, capsules, solutions, emulsions, suspensions, injections, suppositories, sustained-release agents, patches, etc. It can be safely administered orally or parenterally (eg, topical, rectal, intravenous administration, continuous intravenous administration, etc.).
The content of the GRK inhibitor in the pharmaceutical composition is about 0.01 to 11% by weight, preferably about 2 to 85% by weight of the total composition.
The dose of the GRK inhibitor varies depending on the administration subject, administration route, disease and the like. For example, when administered as an oral agent to an adult (body weight of about 60 kg), the GRK inhibitor is about 1 to 3 as the active ingredient GRK inhibitor. 1000 mg, preferably about 3 to 300, more preferably about 10 to 200, which can be administered in 1 to several divided doses per day. These can be appropriately adjusted individually depending on the strength of action, molecular weight, etc. of the GRK inhibitor used.
Further, the `` cardiotonic drug consisting of a GRK inhibitor '' of the present invention is appropriately a drug usually used for treating heart failure, such as digitalis, catecholamine (e.g., dobutamine, dopamine, denopamine, zamoterol, etc.), β-blocker (bisoprolol, Carvedilol), nitrates (nitroglycerin, etc.), hydralazine, Ca antagonists (amlodipine, etc.), ACE inhibitors (enalapril, etc.), AII antagonists (candesartan, etc.), diuretics (furosemide, etc.), PDE inhibitors (milrinone) ), Ca sensitizers (pimobendan, etc.), thrombolytic drugs (t-PA, etc.), anticoagulants (heparin, warfarin, etc.), antiplatelet drugs (aspirin, etc.), antiarrhythmic drugs (amiodarone, etc.), HMG- CoA reductase inhibitors (atorvastatin, simvastatin, etc.), alpha blockers (prazosin, etc.), atrial diuretic peptides, NEP inhibitors (fafactolyl, etc.), endothelin antagonists ( Coal preparation, etc.), aldosterone antagonists (spironolactone, etc.), vasopressin antagonist (conivaptan and the like), matrix metalloproteinase inhibitors, in combination with an equal, or may be used by appropriately blending these pharmaceutical ingredient. Such a combination can be expected to have a synergistic effect as compared to administration alone. Furthermore, it can be used in combination with non-drug therapy for severe heart failure, for example, assisted circulation (balloon pumping in the aorta, auxiliary artificial heart, etc.), batista, heart transplantation and the like.

  Furthermore, the “cardiotonic drug comprising a GRK inhibitor” of the present invention can be used in combination with a biologic (eg, antibody, vaccine preparation, etc.) when applied to each of the above diseases, and a gene therapy method It is also possible to apply as a combination therapy in combination with the above. Examples of antibodies and vaccine preparations include, for example, vaccine preparations for angiotensin II, vaccine preparations for CETP, CETP antibodies, antibodies to TNFα antibodies and other cytokines, amyloid β vaccine preparations, type 1 diabetes vaccines (Peptor's DIAPEP-277, etc.) In addition to antibodies, vaccine preparations against cytokines, renin-angiotensin enzymes and their products, antibodies or vaccine preparations against enzymes and proteins involved in blood lipid metabolism, enzymes and proteins involved in blood coagulation / fibrinolysis systems Examples include antibodies or vaccines, antibodies against vaccines, proteins against vaccine metabolism, insulin metabolism, and the like. In addition, as gene therapy methods, for example, therapeutic methods using genes related to cytokines, renin-angiotensin enzymes and their products, and genes related to signal transduction systems such as β receptor and adenylate cyclase are used. Related treatments, treatment methods using genes related to GRK such as βARKct and βarrestin, treatment methods using DNA decoys such as NFκB decoy, treatment methods using antisense, and enzymes and proteins involved in blood lipid metabolism Involved in angiogenesis therapy for peripheral vascular occlusion, such as treatment methods using genes (eg, genes related to metabolism, excretion or absorption of cholesterol, triglycerides, HDL-cholesterol or blood phospholipids) Therapy using genes related to enzymes and proteins (eg, growth factors such as HGF, VEGF, etc.), sugar metabolism and Treatment using a gene relating to proteins involved in phosphorus resistance, antisense, and the like for the cytokines TNF, and the like. In addition, various organ regeneration methods such as heart regeneration, kidney regeneration, pancreas regeneration, blood vessel regeneration, bone marrow cells (bone marrow mononuclear cells, bone marrow stem cells, etc.) and other cells that have the ability to differentiate into muscle (embryonic stem cells, myoblasts, etc.) It is also possible to use in combination with angiogenesis and myocardial neovascularization using transplantation.

  Examples of administration forms include (1) administration of a single preparation obtained by simultaneously formulating a GRK inhibitor and a concomitant drug, and (2) obtained by separately formulating a GRK inhibitor and a concomitant drug. Co-administration of the same formulation by the same route of administration, (3) administration of the two formulations obtained by separately formulating the GRK inhibitor and the concomitant drug at different times in the same route of administration, (4) Simultaneous administration of two preparations obtained by separately formulating a GRK inhibitor and a concomitant drug by different administration routes, (5) Two preparations obtained by separately formulating a GRK inhibitor and a concomitant drug (For example, administration in the order of the concomitant drugs after administration of the GRK inhibitor, or administration in the reverse order). The dose of the concomitant drug can be appropriately selected based on the clinically used dose. The mixing ratio between the GRK inhibitor and the concomitant drug can be appropriately selected depending on the administration subject, administration route, target disease, symptom, combination and the like. For example, when the administration subject is a human, the concomitant drug may be used in an amount of 0.01 to 100 parts by weight per 1 part by weight of the GRK inhibitor.

  The pharmacological test results using an example of the agent of the present invention are shown below. In addition, this Example is a mere illustration and does not limit the scope of the present invention at all.

３−［［[４−メチル−５−（４−ピリジル）−４Ｈ−１，２，４−トリアゾール−３−イル]メチル］アミノ］安息香酸（１５５ ｍｇ）、２−（トリフルオロメチル）ベンジルアミン（１７５ ｍｇ）、ＷＳＣ（１４４ ｍｇ）および１−ヒドロキシ−１Ｈ−ベンゾトリアゾール一水和物（７．７ ｍｇ）をＤＭＦ（２．０ mL）に混合させ、室温で１８時間撹拌した。反応混合物に酢酸エチル（１００ mL）を加え混合物を水（２×３０ mL）ついで飽和食塩水（２０ mL）で洗浄し、有機層を無水硫酸ナトリウムで乾燥後、溶媒を減圧下留去した。残渣をシリカゲルカラムクロマトグラフィー（酢酸エチル→酢酸エチル：メタノール＝２：１）で精製し題記化合物２２５ ｍｇ（収率９６％）を無色非結晶性物質として得た。
¹H-NMR (300 MHz, Me₂SO-d₆) δ 3.78 (3H, s), 4.55 (2H, d, J = 5.7 Hz), 4.64 (2H, d, J = 5.4 Hz), 6.50 (1H, t, J = 5.4 Hz), 6.95 (1H, d, J = 6.6 Hz), 7.16 (1H, d, J = 7.5 Hz), 7.22 (1H, t, J = 7.8 Hz), 7.29 (1H, s), 7.47 (1H, t, J = 7.8 Hz), 7.51 (1H, d, J = 7.8 Hz), 7.66 (1H, t, J = 7.7 Hz), 7.72-7.75 (3H, m), 8.76 (2H, dd, J = 5.3, 0.8 Hz), 8.94 (1H, t, J = 5.7 Hz). Anal. Calcd. for C₂₄H₂₁F₃N₆O・1.2H₂O: C, 59.06; H, 4.83; N, 17.22. Found: C, 59.16; H, 4.84; N, 16.84. Reference example 1
A method for producing the test compound (1) used in the pharmacological test is shown below.
3-[[[4-Methyl-5- (4-pyridyl) -4H-1,2,4-triazol-3-yl] methyl] amino] -N- [2- (trifluoromethyl) benzyl] benzamide < Test compound (1)

3-[[[4-Methyl-5- (4-pyridyl) -4H-1,2,4-triazol-3-yl] methyl] amino] benzoic acid (155 mg), 2- (trifluoromethyl) benzyl Amine (175 mg), WSC (144 mg) and 1-hydroxy-1H-benzotriazole monohydrate (7.7 mg) were mixed in DMF (2.0 mL) and stirred at room temperature for 18 hours. Ethyl acetate (100 mL) was added to the reaction mixture, and the mixture was washed with water (2 × 30 mL) and then saturated brine (20 mL). The organic layer was dried over anhydrous sodium sulfate, and the solvent was evaporated under reduced pressure. The residue was purified by silica gel column chromatography (ethyl acetate → ethyl acetate: methanol = 2: 1) to give 225 mg (yield 96%) of the title compound as a colorless amorphous substance.
¹ H-NMR (300 MHz, Me ₂ SO-d ₆ ) δ 3.78 (3H, s), 4.55 (2H, d, J = 5.7 Hz), 4.64 (2H, d, J = 5.4 Hz), 6.50 (1H , t, J = 5.4 Hz), 6.95 (1H, d, J = 6.6 Hz), 7.16 (1H, d, J = 7.5 Hz), 7.22 (1H, t, J = 7.8 Hz), 7.29 (1H, s ), 7.47 (1H, t, J = 7.8 Hz), 7.51 (1H, d, J = 7.8 Hz), 7.66 (1H, t, J = 7.7 Hz), 7.72-7.75 (3H, m), 8.76 (2H , dd, J = 5.3, 0.8 Hz), 8.94 (1H, t, J = 5.7 Hz). Anal.Calcd.for C ₂₄ H ₂₁ F ₃ N ₆ O ・ 1.2H ₂ O: C, 59.06; H, 4.83 ; N, 17.22. Found: C, 59.16; H, 4.84; N, 16.84.

According to the test examples described below, the test compound (1) which has substantially only a GRK inhibitory action and was confirmed to show a GRK inhibitory action when applied from outside the cells with a cell or organ perfusion sample is directly applied to the heart. It has been demonstrated that the cardiac contraction force was improved by acting on
Therefore, it is clear that the GRK inhibitory action is an essential action for improving the cardiac contraction force, and it was demonstrated that the GRK inhibitor is useful as a cardiotonic drug.

Test example 1
The following experiment was conducted for the purpose of evaluating the kinase inhibitory action of the test compound (1) on GRK2, PKCα, PKA and ROKα at the enzyme level.
The enzyme inhibitory activity against GRK2 was measured using tubulin as a substrate with reference to a report by Pitcher et al. (Journal of Biological Chemistry, Vol. 273, pages 12316-12324). That is, 0.15 μM purified bovine GRK2, 0.9 μM purified bovine tubulin, 0.5 μM ATP (including 0.1 μCi [γ- ³² P] ATP), 2 mM EDTA, 10 mM MgCl ₂ , 1 mM DTT A test compound having an appropriate concentration was added to 20 mM Tris-HCl (pH 7.5) to prepare a 25 μL mixed solution, which was reacted at 30 ° C. for 1 hour. The reaction was stopped by adding 20% trichloroacetic acid solution (TCA), and then the generated TCA precipitate was collected on a GF / C filter. The radioactivity on the filter was measured using a scintillation counter and used as an index of the amount of tubulin phosphorylation. Normalize the amount of tubulin phosphorylation with the count in the absence of GRK2 as 0% and the count in the absence of the test compound as 100%, and calculate the concentration of the test compound exhibiting 50% phosphorylation inhibition as the IC ₅₀ value. did. Enzyme inhibitory activity against PKCα, PKA and ROKα is 6 μg / mL purified human PKCα or 8 μg / mL purified bovine PKA catalytic subunit as an enzyme, or 0.4 U / mL purified rat ROKα as a substrate at 20 μg / mL. The purified bovine myelin basic protein was measured in the same manner as described above. The results are shown in Table 1.

Table 1 Kinase inhibitory action of test compound (1) on GRK2, PKCα, PKA, ROKα
Kinase IC ₅₀ value
GRK2 35nM
PKCα> 2000nM
PKA> 2000nM
ROKα> 2000nM

From the results in Table 1, it can be seen that the compound (1) exhibits a selective GRK inhibitory action.

Test example 2
The following experiment was conducted for the purpose of evaluating the affinity of the test compound (1) for the adrenergic β receptor, dopamine receptor, endothelin receptor, calcium channel (CaCh) and sodium channel (NaCh). That is, a membrane specimen was prepared from Rex 16 cells highly expressing human β ₁ receptor, and 50 mM Tris-HCl containing 1.4 mM ascorbic acid, 0.001% bovine serum albumin, 1.5 mM CaCl ₂ , 5 mM EDTA, 120 mM NaCl. Incubated with 0.03 nM ¹²⁵ I-Cyanopindolol at 25 ° C. for 2 hours in pH 7.4. Test compound (1) was dissolved in DMSO (final concentration 1%) and added to a final concentration of 10 μM. After the incubation, the reaction solution was filtered through a GF / B filter, and the radioactivity on the filter was measured using a scintillation counter. Nonspecific binding was defined as the amount of binding in the presence of 100 μM S (-)-Propranolol, and the inhibition rate relative to the amount of specific binding in the absence of the test compound was determined. beta ₂ receptor, D ₁ receptors, D _2L receptors, D _2S receptor, D ₃ receptor, D ₅ receptor, ET _A receptor, ET _B receptors, CACH and similar methods also affinity for NaCh Measured with The source of the specimen, respectively, human _{β 2, D 1, D 2L} , D 2S, D 3, D 5, ET A and ET _B receptors with high expressed CHO cells or rat cerebral the (CACH and Nach) using The labeled ligands are 0.2 nM ³ H-CGP-12177 (β ₂ ), 1.4 nM ³ H-SCH-23390 (D ₁ ), 0.16 nM ³ H-Spiperone (D _2L , D _2S ), 0.7 nM ³ H, respectively. -Spiperone (D ₃ ), 2nM ³ H-SCH-23390 (D ₅ ), 0.03 nM ¹²⁵ I-Endothelin-1 (ET _A ), 0.1 nM ¹²⁵ I-Endothelin-1 (ET _B ), 2 nM 3H-Diltiazem ( CaCh) and 5 nM 3H-Batrachotoxin (NaCh) were used. Further, the incubation conditions, respectively 25 ° C. 1 _{h, (β 2), 37 ℃} 2 hours _{(D 1, D 3, D} 5, ET A), 25 ℃ 2 hours _{(D 2L, D 2S, ET} B), 4 ° C for 3 hours (CaCh) and 37 ° C for 1 hour (NaCh) were used. In addition, non-specific binding is 10 μM ICI-118,551 (β ₂ ), 10 μM (+)-Butaclamol (D ₁ ), 10 μM Haloperidol (D _2L , D _2S ), 25 μM S (-)-Sulpiride (D ₃ ), respectively. , 10 μM cis-Flupentixol (D ₅ ), 0.1 μM Endothelin-1 (ET _A , ET _B ), 10 μM Diltiazem (CaCh) and 100 μM Veratridine (NaCh). The results are shown in Table 2.

Table 2 Affinities of test compound (1) for adrenergic β receptor, dopamine receptor and endothelin receptor
Receptor inhibition rate
β ₁ <5%
β ₂ <5%
D ₁ <5%
D _2L <5%
D _2S <5%
D ₃ <5%
D ₅ <5%
ET _A <5%
ET _B <5%
CaCh 6%
NaCh <5%

From the results in Table 2, it can be seen that the test compound (1) has no affinity for various receptors and ion channels involved in the contractility of the myocardium.

Test example 3
The following experiment was conducted for the purpose of evaluating the inhibitory activity of the test compound (1) against Na / K-ATPase and phosphodiesterase (PDE). That is, Na / K-ATPase prepared from canine kidney was preincubated for 15 minutes at 37 ° C in 80 mM Tris-HCl (pH 7.4) containing 1.3 mM EDTA, 25 mM KCl, 5.3 mM MgCl ₂ and 160 mM NaCl. Enzymatic reaction was started by adding 2 mM ATP. Test compound (1) was dissolved in DMSO (final concentration 1%) and added to a final concentration of 10 μM. After a reaction time of 15 minutes, the reaction was stopped by adding 2.5N HClO ₄ and the reaction was caused by adding (NH ₄ ) ₆ Mo ₇ O ₂₄ · H ₂ SO ₄ and Fiske-Subbarow reagent. Pi was colorimetrically determined and used as an indicator of enzyme activity. The inhibition rate of the compound was calculated with the enzyme activity in the absence of the test compound as 100%. Moreover, the inhibitory activity with respect to PDE was measured as follows. In other words, PDE3 prepared from human platelets was preincubated in 10 mM Tris-HCl (pH 7.5) containing 1 mM MgCl ₂ at 25 ° C. for 15 minutes, and then 1 μM cAMP containing ³ H-cAMP was added to start the enzyme reaction. did. Test compound (1) was dissolved in DMSO (final concentration 1%) and added to a final concentration of 10 μM. After 20 minutes of reaction time, stop the reaction by heating at 100 ° C for 2 minutes, add snake venom and incubate at 37 ° C for 10 minutes to convert 3H-AMP produced by the enzyme reaction to 3H-Adenosine. Converted. After removing unreacted cAMP with AG1-X2 resin, the radioactivity in the reaction solution was measured and used as an indicator of enzyme activity. The inhibition rate of the compound was calculated with the enzyme activity in the absence of the test compound as 100%. The enzyme activity of PED4 and PDE5 is 1 μM containing 3H-cGMP instead of 3H-cAMP / cAMP in the case of PED5 assay using human U931 cells (PDE4) and human platelets (PDE5) as the enzyme source. The same measurement was performed using cGMP.
The results are shown in Table 3.

Table 3 Enzyme inhibitory activity of test compound (1) on Na / K-ATPase and phosphodiesterase
Enzyme inhibition rate
Na / K-ATPase <5%
PDE3 <5%
PDE4 <5%
PDE5 <5%

From the results in Table 3, it can be seen that test compound (1) has no enzyme inhibitory activity against Na / K-ATPase and PDE involved in myocardial contractility.

Test example 4
The following experiment was conducted for the purpose of evaluating the GRK inhibitory action in the cell line of the test compound (1).
From the study using βARKct, a GRK inhibitor protein, the accumulation of intracellular cAMP caused by isoproterenol at a concentration that causes sufficient β-receptor desensitization reaction is suppressed by suppressing the GRK-dependent desensitization reaction. (Journal of Clinical Investment, Vol. 99, pp. 288-296). Therefore, we evaluated the potentiating effect on cAMP accumulation after isoproterenol stimulation in HEK293 cells (HEK-B2 cells) with high expression of human β ₂ receptor, and used it as an index of GRK inhibitory action in cell lines. That is, various concentrations of compounds in HEK-B2 cells suspended in D-PBS (+) solution containing 3-isobutyl-1-methyl-xantine (IBMX, 1 mM) and L-ascorbic acid (0.01%) And incubated at 37 ° C. for 20 minutes. Then, a sufficient amount of isoproterenol (1 nM) that causes β receptor desensitization was added and incubated at 37 ° C for 20 minutes to induce cAMP production under desensitization conditions (assay volume 20 μL). ). The cAMP production reaction was stopped by adding the same volume of Tris-EDTA buffer (pH 8.0) and heating at 95 ° C. The amount of cAMP produced was measured using an ALPHA-Screen cAMP detection kit (Perkin Elmer). As a control, the same reaction was performed by adding only a solvent not containing the test compound (N, N-dimethylformamide, final concentration 0.3%). The results are shown by normalizing the amount of cAMP accumulated with the amount of cAMP accumulated in the absence of the compound as 100%. The results are shown in Table 4.

Table 4. Enhancement effect of test compound (1) on cAMP accumulation after isoproterenol stimulation in HEK293 cells highly expressing human β ₂ receptor
Compound concentration cAMP accumulation
1μM 9%
3μM 24%
10μM 52%
30μM 137%

From the results in Table 4, it can be seen that the test compound (1) exhibits a concentration-dependent cAMP accumulation enhancing action, that is, a GRK inhibitory action, when applied extracellularly.

Test Example 5
In order to confirm that the β receptor desensitization inhibitory effect by GRK inhibition of the test compound (1) confirmed in Test Example 4 also occurs at the heart organ level, the following experiment was conducted using a rat isolated perfused heart specimen. went.
The heart of a 10-11 week old male Wistar rat was removed, a catheter was inserted into the aortic root, and it was attached to a Langendorff apparatus kept at 37 ° C, and mixed gas (95% O ₂ -5% CO ₂ ) A Krebs solution (composition (mM): NaCl 113, KCl 4.6, CaCl ₂ 1.2, NaH ₂ PO ₄ 2.5, MgCl ₂ 1.2, NaHCO ₃ 22, glucose 10) aerated with a constant pressure was perfused (75 mmHg). After the stabilization period, 10 μM of solvent (N, N-dimethylformamide, final concentration 0.1%) or compound (1) is applied for 20 minutes, followed by 10 μM isoproterenol (Iso) to cause β-receptor desensitization. ) Was applied for an additional 20 minutes. Isoproterenol was not applied in the control group that did not cause β receptor desensitization. The heart after drug application was frozen in liquid nitrogen and stored at −80 ° C. until the following experiment. The preparation of the pericardial membrane preparation was in accordance with the method of Drazner et al. (Journal of Clinical Investigation, Vol. 99, pp. 288-296). That is, the heart was thawed in lysis buffer (25 mM Tris-HCl, 5 mM EDTA, 5 mM EGTA, pH 7.4 at 37 ° C), homogenized, and centrifuged at 500 xg to obtain the nuclear fraction. The supernatant was centrifuged at 40,000 × g to precipitate the membrane fraction. The obtained membrane fraction was resuspended in a binding buffer (75 mM Tris-HCl, 12.5 mM MgCl ₂ , 2 mM EDTA, pH 7.4 at 37 ° C.). The β-receptor reactivity of membrane specimens was measured using isoproterenol-dependent adenylate cyclase (AC) activity as an index. In other words, the membrane sample is suspended in the presence of 0.5 mM IBMX, 53 μM GTP, 2.7 mM pyruvate, 4 IU / mL pyruvate kinase, 20 IU / mL myokinase, solvent, 0.3 μM isoproterenol or 10 mM sodium fluoride (NaF). CAMP was produced by adding 0.12 mM ATP in the presence of and incubating at 37 ° C. The reaction was stopped by addition of trichloroacetic acid, and after deproteinization and TCA removal, the produced cAMP was measured using an ALPHA-Screen cAMP detection kit. The results were normalized with the amount of cAMP obtained in the presence of NaF as 100%, and were shown as the average value and standard error of 8 cases. The results are shown in Table 5.

Table 5. Inhibitory effect of compound (1) on β-receptor responsiveness reduction (desensitization) by application of high-concentration isoproterenol in isolated rat perfused heart preparations
AC activity
Treatment (% of 10mM NaF)
Control 5.7 ± 0.6%
Solvent + Iso 3.3 ± 0.6%
Test compound (1) + Iso 5.3 ± 0.2%

As shown in Table 5, the decrease in β-receptor reactivity (desensitization) caused by application of high-concentration isoproterenol in rat isolated perfused heart specimens was significantly suppressed by pretreatment with 10 μM test compound (1). This indicates that the β receptor desensitization inhibitory effect by GRK inhibition of the test compound (1) confirmed in Test Example 4 also occurs at the heart organ level.

Test Example 6
The following experiment was conducted for the purpose of observing the effect of acute GRK inhibition using the test compound (1) on the cardiac contractility of the isolated perfused heart specimen.
The heart of a 10-11 week old male Wistar rat was removed, a catheter was inserted into the aortic root, and it was attached to a Langendorff apparatus kept at 37 ° C, and mixed gas (95% O ₂ -5% CO ₂ ) A Krebs solution (composition (mM): NaCl 113, KCl 4.6, CaCl ₂ 1.2, NaH ₂ PO ₄ 2.5, MgCl ₂ 1.2, NaHCO ₃ 22, glucose 10) aerated with a constant pressure was perfused (75 mmHg). The left ventricular pressure, which is an index of cardiac contraction force, was measured from a balloon catheter inserted from the left atrial appendage through a pressure transducer. The heart rate was measured from the left ventricular pressure via a tachometer (Nihon Kohden). After the stabilization period, the compound (1) dissolved in N, N-dimethylformamide was mixed with the perfusate at a ratio of 0.1% and applied. After 15 minutes of drug perfusion, 1 nM isoproterenol dissolved in distilled water was simultaneously perfused for 15 minutes to examine the action of compound (1) in the presence of catecholamine. As a control, only the solvent was perfused for 15 minutes, and then 1nM isoproterenol was used in combination for 15 minutes, and the same observation was performed. The change in each parameter that occurred when Compound (1) was applied alone and when combined with isoproterenol (+ Iso) was shown as the average of 4-5 cases as the rate of change from the value before drug administration. The results are shown in Table 6.

Table 6 Effect of test compound (1) on cardiac contractile force in isolated rat perfused heart specimens
Left ventricular developed pressure heart rate
Alone + Iso alone + Iso
Solvent 9% 33% 2% 28%
Test compound (1) 1μM 20% 39% 4% 22%
Test compound (1) 10μM 45% 97% 2% 35%

As shown in Table 5, by selectively inhibiting the cardiac GRK with the test compound (1), a cardiac contractile force enhancing effect depending on the compound concentration is exhibited immediately after drug administration, and the effect is under the coexistence of catecholamine. It became clear that it increased further. The working concentration of the test compound (1) was consistent with the effective concentration of the GRK inhibitory action obtained in the cell line. Interestingly, this effect of enhancing cardiac contractility by acute GRK inhibition was hardly accompanied by an increase in heart rate.

Test Example 7
The following experiment was conducted for the purpose of observing the effect of acute GRK inhibition using the test compound (1) on in vivo cardiovascular dynamics.
Male Wistar rats (350-450 g) were anesthetized with inactin (100 mg / kg, ip), and cannulas for blood pressure (MBP) measurement and drug administration were placed in the left femoral artery and vein, respectively. A mirror catheter with a pressure sensor was placed in the left ventricle from the right carotid artery, and the left ventricular pressure (LVP) was measured. By differentiating LVP, the maximum left ventricular pressure first derivative (LVdP / dt _max ), which is an index of cardiac contraction force, and the minimum left ventricular pressure first derivative (LVdP / dt _min ), which is an index of cardiac relaxation, are calculated. did. Heart rate (HR) was measured from the left ventricular pressure via a tachometer. After the stabilization period, the test compound (1) was dissolved in polyethylene glycol (PEG) 400 containing 1N HCl and then diluted with physiological saline to give a 10 mg / mL solution (final PEG concentration 30%, HCl concentration 0.02N). And was administered intravenously at a dose of 1 mg / kg / min for 30 minutes, and cardiovascular changes were recorded until 20 minutes after the completion of administration. A solvent containing no compound (30% PEG / HCl / saline) was similarly administered to the control group. The data obtained in Fig. 1 are shown as the rate of change from the value before the start of drug administration, with the mean value and standard error of 4 cases. As shown in FIG. 1, the GRK was selectively suppressed in vivo by continuous intravenous administration of the test compound (1), just like the results in the isolated perfused heart specimen (Test Example 5). As a result, only the systolic force was enhanced with little effect on blood pressure and heart rate, and the effect was recovered by discontinuation of administration. This result clearly shows that inhibition of GRK is a new mechanism of “cardiotonic drugs” that improve cardiac contraction force acutely.

Formulation Example 1
In the present invention, “a cardiotonic drug containing a GRK inhibitor” can be implemented, for example, in the form of a pharmaceutical composition having the following specific formulation (however, it is not limited thereto). Absent).
In addition, in the following prescription, components (additives) other than the active ingredient may be listed products in the Japanese Pharmacopoeia, the Japanese Pharmacopoeia Standards for Pharmaceuticals or the Standards for Pharmaceutical Additives, and the like.

1. Capsule (1) Test compound (1) 40 mg
(2) Lactose 70mg
(3) Microcrystalline cellulose 9mg
(4) Magnesium stearate 1mg
1 capsule 120mg
After mixing (1), (2) and 1/2 of (3) and (4), granulate. The remaining (4) is added to this and the whole is enclosed in a gelatin capsule.

2. Tablet (1) Test compound (1) 40 mg
(2) Lactose 58mg
(3) Corn starch 18mg
(4) Microcrystalline cellulose 3.5mg
(5) Magnesium stearate 0.5mg
1 tablet 120mg
After mixing 2/3 of (1), (2), (3), (4) and 1/2 of (5), granulate. The remaining (4) and (5) are added to the granules and pressed into tablets.

  It was first demonstrated by the present invention that a GRK inhibitor has an excellent cardiotonic effect. The agent of the present invention comprising a GRK inhibitor is useful as a cardiotonic agent and has extremely high utility value in the field of the pharmaceutical industry.

FIG. 1 shows the results of observing the effect of acute GRK inhibition by the test compound (1) on the cardiovascular dynamics of rats.

Claims (2)

A cardiotonic drug containing a GRK (G protein-coupled receptor kinase) inhibitor. The cardiotonic agent according to claim 1, wherein the GRK inhibitor is a GRK2 inhibitor.

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