WO2024043242A1 - Heart failure treatment via cardiotonic effect by trpc3/6/7 channel activation - Google Patents

Heart failure treatment via cardiotonic effect by trpc3/6/7 channel activation Download PDF

Info

Publication number
WO2024043242A1
WO2024043242A1 PCT/JP2023/030211 JP2023030211W WO2024043242A1 WO 2024043242 A1 WO2024043242 A1 WO 2024043242A1 JP 2023030211 W JP2023030211 W JP 2023030211W WO 2024043242 A1 WO2024043242 A1 WO 2024043242A1
Authority
WO
WIPO (PCT)
Prior art keywords
group
atom
substituted
alkoxycarbonyl
groups
Prior art date
Application number
PCT/JP2023/030211
Other languages
French (fr)
Japanese (ja)
Inventor
基宏 西田
和宏 西山
百合 加藤
明幸 西村
龍 永田
泰生 森
靖章 中川
宏一郎 桑原
Original Assignee
国立大学法人九州大学
大学共同利用機関法人自然科学研究機構
国立大学法人大阪大学
国立大学法人京都大学
国立大学法人信州大学
Priority date (The priority date is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the date listed.)
Filing date
Publication date
Application filed by 国立大学法人九州大学, 大学共同利用機関法人自然科学研究機構, 国立大学法人大阪大学, 国立大学法人京都大学, 国立大学法人信州大学 filed Critical 国立大学法人九州大学
Publication of WO2024043242A1 publication Critical patent/WO2024043242A1/en

Links

Classifications

    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K31/00—Medicinal preparations containing organic active ingredients
    • A61K31/045—Hydroxy compounds, e.g. alcohols; Salts thereof, e.g. alcoholates
    • A61K31/05—Phenols
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K31/00—Medicinal preparations containing organic active ingredients
    • A61K31/33—Heterocyclic compounds
    • A61K31/395—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins
    • A61K31/435—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins having six-membered rings with one nitrogen as the only ring hetero atom
    • A61K31/44—Non condensed pyridines; Hydrogenated derivatives thereof
    • A61K31/445—Non condensed piperidines, e.g. piperocaine
    • A61K31/4523—Non condensed piperidines, e.g. piperocaine containing further heterocyclic ring systems
    • A61K31/454—Non condensed piperidines, e.g. piperocaine containing further heterocyclic ring systems containing a five-membered ring with nitrogen as a ring hetero atom, e.g. pimozide, domperidone
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K31/00—Medicinal preparations containing organic active ingredients
    • A61K31/33—Heterocyclic compounds
    • A61K31/395—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins
    • A61K31/495—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins having six-membered rings with two or more nitrogen atoms as the only ring heteroatoms, e.g. piperazine or tetrazines
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K31/00—Medicinal preparations containing organic active ingredients
    • A61K31/33—Heterocyclic compounds
    • A61K31/395—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins
    • A61K31/495—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins having six-membered rings with two or more nitrogen atoms as the only ring heteroatoms, e.g. piperazine or tetrazines
    • A61K31/496—Non-condensed piperazines containing further heterocyclic rings, e.g. rifampin, thiothixene or sparfloxacin
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K45/00—Medicinal preparations containing active ingredients not provided for in groups A61K31/00 - A61K41/00
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P21/00—Drugs for disorders of the muscular or neuromuscular system
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P9/00—Drugs for disorders of the cardiovascular system
    • A61P9/04—Inotropic agents, i.e. stimulants of cardiac contraction; Drugs for heart failure
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07D—HETEROCYCLIC COMPOUNDS
    • C07D401/00—Heterocyclic compounds containing two or more hetero rings, having nitrogen atoms as the only ring hetero atoms, at least one ring being a six-membered ring with only one nitrogen atom
    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N33/00—Investigating or analysing materials by specific methods not covered by groups G01N1/00 - G01N31/00
    • G01N33/15—Medicinal preparations ; Physical properties thereof, e.g. dissolubility
    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N33/00—Investigating or analysing materials by specific methods not covered by groups G01N1/00 - G01N31/00
    • G01N33/48—Biological material, e.g. blood, urine; Haemocytometers
    • G01N33/50—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing

Definitions

  • the present invention generally relates to new uses for TRPC3/6/7 channel activators.
  • the present invention particularly relates to pharmaceutical compositions containing TRPC3/6/7 channel activators, specifically pharmaceutical compositions for preventing and/or treating heart failure or skeletal muscle failure, and
  • the present invention relates to a method for screening substances for preventing and/or treating disorders.
  • heart failure has been increasing around the world as society ages, and a situation that can be called a "heart failure pandemic" is progressing.
  • the number of heart failure patients worldwide is on track to reach 30 million, and there are concerns that this number will continue to increase, leading to a shortage of hospital beds.
  • heart failure is a pathological condition with a poor prognosis, with no complete recovery, and a gradual decline in heart function (chronic heart failure) through repeated periods of acute exacerbation (acute heart failure) and remission (chronic heart failure), eventually leading to treatment-resistant heart failure and death.
  • chronic heart failure a pathological condition with a poor prognosis, with no complete recovery, and a gradual decline in heart function (chronic heart failure) through repeated periods of acute exacerbation (acute heart failure) and remission (chronic heart failure), eventually leading to treatment-resistant heart failure and death.
  • acute heart failure acute heart failure
  • remission chronic heart failure
  • the pumping activity and morphological structure of the heart are regulated by various neurohumoral factors.
  • the cells that make up the heart maintain cardiac homeostasis by sensing these neurohumoral factors on their cell membranes and transmitting appropriate signals within the cells.
  • Cardiomyocyte hypertrophy and interstitial fibrosis that occur with the onset and progression of heart failure are caused by Ca 2+ signaling, especially Ca 2+
  • the influx of Ca 2+ from outside the cell into the cell is deeply involved in the induction of expression of genes related to hypertrophy, and transient Ca 2+ influx is attracting attention as a molecular entity responsible for receptor-mediated Ca 2+ influx. It is a receptor potential canonical (TRPC) protein.
  • TRPC receptor potential canonical
  • TRPC1-TRPC7 There are seven isoforms (TRPC1-TRPC7) of animal TRPC channels, and among them, lipid (diacylglycerol)-operated TRPC3 and TRPC6 channels are important for cardiac remodeling, including the results of the present applicant. There have been many reports. In fact, it has been demonstrated at the animal level that compounds that inhibit TRPC3/C6 channel activity improve pathological cardiac hypertrophy and heart failure, suggesting their effectiveness as therapeutic targets (Non-patent Documents 1-7 ). Further, Patent Document 1 describes a benzisoxazole compound that modulates or inhibits the activity of TRPC3 channel or TRPC6 channel, and mentions cardiac hypertrophy as a TRPC3 and/or TRPC6-related disease. Substances that activate the channels are not described. In any case, the physiological role of TRPC3/6 channels in regulating cardiac function is not well understood.
  • Kiyonaka S Kato K, Nishida M, Mio K, Numaga T, Sawaguchi Y, Yoshida T, Wakamori M, Mori E, Numata T, Ishii M, Takemoto H, Ojida A, Watanabe K, Uemura A, Kurose H, Morii T , Kobayashi T, Sato Y, Sato C, Hamachi I, Mori Y Selective and direct inhibition of TRPC3 channels underlies biological activities of a pyrazole compound. Proc. Natl. Acad. Sci. USA. 106: 5400-5405 (2009).
  • therapeutic drugs to treat heart failure a condition with a poor prognosis
  • TRPC6 gene-deficient mice to be vulnerable to heart failure, that is, the heart of TRPC6-deficient mice was unable to adapt to high blood pressure load, and activated TRPC3/6/7 channels.
  • TRPC3/6/7 channels We attempted to apply substances that cause disease to pathological conditions.
  • substances that activate TRPC3/6/7 channels have the same protective effect on heart failure even in different heart failure models and different mouse strains, and have completed the present invention.
  • a pharmaceutical composition for preventing and/or treating heart failure or skeletal muscle failure containing a TRPC3/6/7 channel activator.
  • the TRPC3/6/7 channel activator has the formula (1): [In the formula, A represents a carbon atom or a nitrogen atom, B represents a carbon atom or a nitrogen atom, R 1 represents an optionally substituted alkyl group, and when there is a plurality of them, they may be the same or different, m represents an integer from 0 to 5, R 2 is a halogen atom; a trihalogenomethyl group; an alkyl group; an acyloxy group; an alkoxycarbonyl group; an optionally substituted amide group; an optionally substituted piperidinylcarbonyl group; or an optionally substituted pyrrolidinyl Indicates a carbonyl group, and when there are multiple groups, they may be the same or different, n represents an integer
  • the TRPC3/6/7 channel activator has the formula (1): [In the formula, A represents a carbon atom or a nitrogen atom, B represents a carbon atom or a nitrogen atom, R 1 represents a halogen atom, an alkyl group, an alkoxy group, an acyloxy group, an alkoxycarbonyl group, or an alkyl group substituted with a halogen atom, and when there is more than one, they may be the same or different; m represents an integer from 0 to 5, R 2 may be substituted with one or two groups selected from the group consisting of a halogen atom; a trihalogenomethyl group; an alkyl group; an acyloxy group; an alkoxycarbonyl group; an alkyl group, an alkoxycarbonyl group, and a halogen atom.
  • a good amide group; a piperidinylcarbonyl group which may be substituted with one or more groups selected from the group consisting of an alkyl group, an alkoxycarbonyl group, and a halogen atom; or an alkyl group, an alkoxycarbonyl group, and a halogen atom Indicates a pyrrolidinylcarbonyl group which may be substituted with one or more groups selected from the group consisting of, and when there are multiple groups, they may be the same or different, n represents an integer from 0 to 5, p represents 0 or 1, and q represents 0 or 1.
  • the TRPC3/6/7 channel activator is a compound represented by the formula (1), A represents a carbon atom or a nitrogen atom, B represents a carbon atom or a nitrogen atom, R 1 represents a fluorine atom, a chlorine atom, a bromine atom, a C1-C4 alkyl group, a C1-C4 alkoxy group, a C2-C5 acyloxy group, a C2-C5 alkoxycarbonyl group, or a trihalogenomethyl group, and when there is more than one, may be the same or different; m represents an integer from 0 to 4, R 2 is a fluorine atom; a chlorine atom; a bromine atom; a trifluoromethyl group; a trichloromethyl group; a tri
  • the TRPC3/6/7 channel activator is a compound represented by the formula (1), A represents a nitrogen atom, B represents a carbon atom or a nitrogen atom, R 1 represents a fluorine atom, a chlorine atom, a C1-C4 alkyl group, a C1-C4 alkoxy group, a C2-C4 alkoxycarbonyl group, a trifluoromethyl group, a trichloromethyl group, or a tribromomethyl group, and when there is more than one, may be the same or different; m represents an integer from 0 to 3, R 2 is a fluorine atom; a chlorine atom; a trifluoromethyl group; a trichloromethyl group; a C1 to C4 alkyl group; a C2 to C4 acyloxy group; a C2 to C4 alkoxycarbonyl group; a C1 to C3 alkyl group and a C2 to C4 alkoxycarbonyl group
  • the TRPC3/6/7 channel activator is a compound represented by the formula (1), A represents a nitrogen atom, B represents a carbon atom or a nitrogen atom, R 1 represents a chlorine atom, a C1-C3 alkyl group, a C1-C3 alkoxy group, a C2-C4 alkoxycarbonyl group, a trifluoromethyl group, or a trichloromethyl group, and when there is more than one, they may be the same or different, m represents an integer from 0 to 2, R 2 is selected from the group consisting of a fluorine atom; a chlorine atom; a trifluoromethyl group; a trichloromethyl group; a C1-C4 alkyl group; a C2-C4
  • an amide group which may be substituted with one or two groups; or substituted with one or more groups selected from the group consisting of a C1-C3 alkyl group, a C2-C4 alkoxycarbonyl group, and a halogen atom; Indicates a piperidinyl carbonyl group that may be n represents an integer from 0 to 3; p indicates 1, q indicates 1, The pharmaceutical composition according to [5].
  • ⁇ Screening method> [9] A method of screening for substances for preventing and/or treating heart failure or skeletal muscle failure, the screening method comprising selecting TRPC3/6/7 channel activators. [10] measuring the zinc concentration in cells in the presence of a test substance, and if the zinc concentration is increased compared to the zinc concentration in the absence of the test substance, the test substance.
  • A represents a carbon atom or a nitrogen atom
  • B represents a carbon atom or a nitrogen atom
  • R 1 represents a halogen atom, an alkyl group, an alkoxy group, an acyloxy group, an alkoxycarbonyl group, or an alkyl group substituted with a halogen atom, and when there is more than one, they may be the same or different
  • m represents an integer from 0 to 5
  • n represents an integer from 2 to 5
  • At least one of the plurality of R 2 is a halogen atom, an alkyl group, or a trifluoromethyl group; the remaining R 2 are 1) substituted with a single or multiple halogen atoms, and are selected from an alkyl group and an alkoxycarbonyl group.
  • Figure 1 is a graph showing the increase in left ventricular contractility and heart rate due to intravenous administration of the vasodilator hydralazine (Hyd) in wild type (WT) and TRPC6 gene-deficient mice (TRPC6(-/-)). It is. ⁇ LVdP/dtmax: increase in left ventricular contractile force, ⁇ Heart Rate: increase in heart rate, AUC: area under the curve (total area under the curve for 10 minutes after administration). **; P ⁇ 0.01.
  • Figure 2 shows the inhibitory effect of PPZ2 on the progression of heart failure in MLP-deficient mice.
  • Figure 2a Graph showing left ventricular ejection fraction (EF).
  • Figure 2b A graph showing fractional shortening (FS) of the left ventricle.
  • Figure 3-1 shows the heart failure improving effect of PPZ2 in mice with pressure overload-induced heart failure.
  • Figure 3-1a A graph showing left ventricular ejection fraction (EF).
  • Figure 3-2 shows the heart failure ameliorating effect of PPZ2 in mice with pressure overload-induced heart failure.
  • FIG. 1a A graph showing left ventricular ejection fraction (EF).
  • Figure 3-1c A photograph of a WGA-stained image (left) and a graph
  • Figure 3-2e Photograph showing Zn 2+ concentration (Zinpyr-1 staining) in myocardial tissue.
  • FIG. 5 is a graph showing the dose dependence of enhancement of the baroreceptor reflex response (positive inotropic effect) in normal mice by administration of PPZ2 and the suppressive effect of the TRPC6 inhibitor BI749327.
  • the vertical axis shows the hydralazine-induced increase in left ventricular contractile force ( ⁇ LVdP/dtmax) or heart rate ( ⁇ Heart Rate).
  • the ⁇ value indicates the difference between the value before hydralazine (Hyd) administration and the maximum value 10 minutes after administration.
  • FIG. 6 shows changes in intracellular Zn 2+ concentration caused by TRPC6 activators using TRPC6-expressing HEK293 cells.
  • Figures 6a and 6b are graphs comparing the FluoZn-3 fluorescence intensity change and its increase rate (AUC; area under the curve) due to PPZ2 (30 ⁇ M) treatment.
  • FIG. 7 shows the results of examining the myocardial contractility enhancing effect of TRPC6 activators using neonatal rat cardiomyocytes.
  • Figure 7a Showing the effect of PPZ2 (30 ⁇ M) treatment on cardiomyocyte contraction induced by norepinephrine (NE) (0.1 ⁇ M) or the zinc chelator TPEN (N,N,N',N'-tetrakis(2-pyridylmethylethylenediamine).
  • Figure 7b Graph showing the effect of GSK1702934A (1 ⁇ M) or hyperforin (1 ⁇ M) treatment on cardiomyocyte contraction induced by NE (1 ⁇ M).
  • FIG. 8 is a graph showing that PPZ2 suppressed the increase in myopathy markers (left: LDH, right: CK) in muscular dystrophy model mice.
  • FIG. 9 is a graph showing the effects of L687 and PPZ2 on Ca 2+ influx into HEK293 cells expressing human TRPC6 channels.
  • FIG. 10 is a graph showing the effect of the compounds of the present invention on Ca 2+ influx into HEK293 cells expressing human TRPC6 channels.
  • FIG. 11 is a graph showing the effect of the compounds of the present invention on Ca 2+ influx into HEK293 cells expressing human TRPC3 channels.
  • Figure 12. Upper panel shows ⁇ -actinin staining images of neonatal rat ventricular myocytes.
  • FIG. 14 is a graph showing the effect of L-0044 (1 ⁇ M) treatment on the contraction of cardiomyocytes stimulated by norepinephrine (NE) (0.1 ⁇ M).
  • the present invention provides that a low-molecular compound that promotes activation of receptor-operated TRPC3/6/7 channels becomes an effective therapeutic agent for acute heart failure or acute exacerbation of chronic heart failure without having a hypotensive effect.
  • This is a use invention showing the following. In a state of heart failure, the ability of the heart to pump blood is originally weakened, making it difficult for blood pressure to rise. When antihypertensive effects are added here, a state of chronic hypotension occurs, and a compensatory mechanism that attempts to raise blood pressure, ie, an increase in sympathetic nerve activity, comes into play.
  • the present invention provides a pharmaceutical composition for preventing and/or treating heart failure or skeletal muscle failure, containing a TRPC3/6/7 channel activator, specifically a piperazine derivative (PPZ). provide.
  • a TRPC3/6/7 channel activator specifically a piperazine derivative (PPZ).
  • TRPC6 gene-deficient mice show a tendency to be vulnerable to heart failure, that is, the hearts of TRPC6-deficient mice are susceptible to increased oxidative stress and inflammation in response to hypertensive stress, and are unable to adapt to hypertensive stress. I paid attention to that.
  • Norepinephrine released from sympathetic nerve terminals is a major neurotransmitter that enhances the pumping function of the heart, and plays an especially important role in compensatory cardiac hyperactivity (baroreceptive reflex) in response to sudden drops in blood pressure.
  • Noradrenaline activates ⁇ -adrenergic receptors on the cardiac muscle cell membrane, causing contraction of the cardiac muscle cell and an increase in heart rate.
  • TRPC6-deficient mice showed positive baroreflex responses (positive chronotropy (increased heart rate) and positive inotropy (increased contractile force)) induced by the antihypertensive drug hydralazine.
  • the inotropic effect was significantly attenuated ( Figure 1).
  • the baroreceptive reflex response is also markedly attenuated in heart failure, which is a cause of worsening prognosis, suggesting that TRPC6 channel activation may work to improve heart failure.
  • the present inventors focused on this relationship and found for the first time a relationship between activation of TRPC3/6/7 channels and prevention or treatment of heart failure or skeletal muscle failure.
  • Test Examples 1-3 described below different experimental animals were used, and despite the fact that the phenotype often changes due to model differences, species differences, and strain differences, different heart failure models and different mouse strains were used. We were able to show that the same protective effect against heart failure exists (commonality/universality).
  • TRPC3/6/7 channel activator refers to a group of channels that are directly activated by diacylglycerol (TRPC3/ 6/7), which activates the opening of diacylglycerol independently of diacylglycerol.
  • TRPC3/6/7 channel activator means a group of substances that activate any or all of TRPC3, TRPC6, and TRPC7.
  • the "TRPC3/6/7 channel activator” includes piperazine derivatives (PPZ), and specific examples thereof include Sawamura S, Hatano M, Takada Y, Hino K, Kawamura T, Tanikawa J, Nakagawa H, Hase H, Nakao A, Hirano M, Rotrattanadumrong R, Kiyonaka S, Mori MX, Nishida M, Hu Y, Inoue R, Nagata R, Mori Y. Screening of Transient Receptor Potential Canonical Channel Activators Identifies Novel Neurotrophic Piperazine Compounds. Examples include piperazine derivatives described in Mol Pharmacol. 89(3), 348-363 (2016). doi: 10.1124/mol.115.102863. For example, the formula: Examples include any of the compounds shown below.
  • TRPC3/6/7 channel activator includes Xu X, Lozinskaya I, Costell M, Lin Z, Ball JA, Bernard R, Behm DJ, Marino JP, Schnackenberg (2013).
  • TRPC3/6/7 channel activator includes cannabidiol, N. Qin et al., which is known to activate TRPV1 channel, TRPV2 channel, and TRPA1 channel. J. Neurosci. 28, 6231-6238 (2008)) is also included.
  • TRPC3/6/7 channel activator includes compounds including the compounds described in WO2022/118966.
  • the present invention provides a pharmaceutical composition for preventing and/or treating heart failure or skeletal muscle failure, which contains a TRPC3/6/7 channel activator;
  • the TRPC3/6/7 channel activator has the formula (1): [In the formula, A represents a carbon atom or a nitrogen atom, B represents a carbon atom or a nitrogen atom, R 1 represents an optionally substituted alkyl group, and when there is a plurality of them, they may be the same or different, m represents an integer from 0 to 5, R 2 is a halogen atom; a trihalogenomethyl group; an alkyl group; an acyloxy group; an alkoxycarbonyl group; an optionally substituted amide group; an optionally substituted piperidinylcarbonyl group; or an optionally substituted pyrrolidinyl Indicates a carbonyl group, and when there are multiple groups, they may be the same or different, n represents an integer from 0 to 5, p
  • the TRPC3/6/7 channel activator has the formula (1): [In the formula, A represents a carbon atom or a nitrogen atom, B represents a carbon atom or a nitrogen atom, R 1 represents a halogen atom, an alkyl group, an alkoxy group, an acyloxy group, an alkoxycarbonyl group, or an alkyl group substituted with a halogen atom, and when there is more than one, they may be the same or different; m represents an integer from 0 to 5, R 2 is one or two groups selected from the group consisting of a halogen atom; a trihalogenomethyl group; an alkyl group; an acyloxy group; an alkoxycarbonyl group; an alkyl group, an alkoxycarbonyl group, and a halogen atom, preferably a fluorine atom.
  • the TRPC3/6/7 channel activator is a compound represented by the above formula (1), A represents a carbon atom or a nitrogen atom, B represents a carbon atom or a nitrogen atom, R 1 represents a fluorine atom, a chlorine atom, a bromine atom, a C1-C4 alkyl group, a C1-C4 alkoxy group, a C2-C5 acyloxy group, a C2-C5 alkoxycarbonyl group, or a trihalogenomethyl group, and when there is more than one, may be the same or different; m represents an integer from 0 to 4, R 2 is a fluorine atom; a chlorine atom; a bromine atom; a trifluoromethyl group; a trichloromethyl group; a tribromomethyl group; a C1 to C4 alkyl group; a C2 to C5 acyloxy group; a C2 to C5 alkoxycarbon
  • the TRPC3/6/7 channel activator is a compound represented by the formula (1), A represents a nitrogen atom, B represents a carbon atom or a nitrogen atom, R 1 represents a fluorine atom, a chlorine atom, a C1-C4 alkyl group, a C1-C4 alkoxy group, a C2-C4 alkoxycarbonyl group, a trifluoromethyl group, a trichloromethyl group, or a tribromomethyl group, and when there is more than one, may be the same or different; m represents an integer from 0 to 3, R 2 is a fluorine atom; a chlorine atom; a trifluoromethyl group; a trichloromethyl group; a C1 to C4 alkyl group; a C2 to C4 acyloxy group; a C2 to C4 alkoxycarbonyl group; a C1 to C3 alkyl group and a C2 to C4 alkoxycarbonyl
  • the TRPC3/6/7 channel activator is a compound represented by the formula (1), A represents a nitrogen atom, B represents a carbon atom or a nitrogen atom, R 1 represents a chlorine atom, a C1-C3 alkyl group, a C1-C3 alkoxy group, a C2-C4 alkoxycarbonyl group, a trifluoromethyl group, or a trichloromethyl group, and when there is more than one, they may be the same or different, m represents an integer from 0 to 2, R 2 is selected from the group consisting of a fluorine atom; a chlorine atom; a trifluoromethyl group; a trichloromethyl group; a C1-C4 alkyl group; a C2-C4 alkoxycarbonyl group; a C1-C3 alkyl group and a C2-C4 alkoxycarbonyl group.
  • A represents a nitrogen atom
  • B represents a carbon atom or a nitrogen atom
  • amide group which may be substituted with one or two groups; or one or two selected from the group consisting of a C1-C3 alkyl group, a C2-C4 alkoxycarbonyl group, and a halogen atom, preferably a fluorine atom;
  • A represents a carbon atom or a nitrogen atom
  • B represents a carbon atom or a nitrogen atom.
  • a and B are both carbon atoms
  • A is a carbon atom and B is a nitrogen atom
  • A is a nitrogen atom and B is a carbon atom
  • a and B are both nitrogen atoms. That is, the ring containing A and B in formula (1) is a benzene ring, a pyridine ring, a pyridazine ring, a pyridine ring, or a pyridazine ring.
  • both A and B are carbon atoms, A is a carbon atom and B is a nitrogen atom, or A is a nitrogen atom and B is a carbon atom; It is more preferable that both B are carbon atoms, or A is a nitrogen atom and B is a carbon atom.
  • the carbon atom may have a substituent R 1 .
  • R 1 is preferably a fluorine atom, a chlorine atom, a bromine atom, a C1-C4 alkyl group, a C1-C4 alkoxy group, a C2-C5 acyloxy group, a C2-C5 alkoxycarbonyl group, or a trihalogenomethyl group, and a fluorine atom, A chlorine atom, a C1-C4 alkyl group, a C1-C4 alkoxy group, a C2-C4 alkoxycarbonyl group, a trifluoromethyl group, a trichloromethyl group, or a tribromomethyl group are more preferable, and a chlorine atom, a C1-C3 alkyl group, a C1 -C3 alkoxy group, C2-C4 alkoxycarbonyl group, trifluoromethyl group, or trichloromethyl group is more preferred
  • R 1 is located at the ortho position to the carbon atom directly bonding to the nitrogen atom constituting (NH) p among the carbon atoms constituting the benzene ring. Preferably, it is bonded to at least one of two carbon atoms.
  • R 1 bonded to the carbon atom located at the ortho position include an alkoxy group or an alkoxycarbonyl group, preferably a C2-C5 alkoxycarbonyl group, more preferably a C2-C4 alkoxycarbonyl group, and a methoxycarbonyl group or an ethoxycarbonyl group.
  • a carbonyl group is more preferred, and an ethoxycarbonyl group is particularly preferred.
  • the ring to which R 1 is bonded is a benzene ring, and among the carbon atoms constituting the benzene ring, it is located at the meta position with respect to the carbon atom directly bonded to the nitrogen atom constituting (NH) p .
  • a halogen atom is preferable, a chlorine atom or a fluorine atom is more preferable, and a fluorine atom is particularly preferable.
  • R 1 is the carbon atom that directly bonds to the nitrogen atom that constitutes (NH) p among the carbon atoms that constitute the pyridine ring or pyridazine ring. It is preferable to bond to a carbon atom located between A and A.
  • R 1 may be, for example, a halogen atom, an alkyl group, an alkoxy group, or an alkoxycarbonyl group, and may be a fluorine atom, a chlorine atom, a C1-C4 alkyl A C1-C4 alkoxy group or a C2-C5 alkoxycarbonyl group is preferred, a C1-C4 alkoxy group is more preferred, and a methoxy group or an ethoxy group is particularly preferred.
  • m is preferably an integer of 0 to 4, more preferably an integer of 0 to 3, even more preferably 0, 1, or 2, even more preferably 0 or 1, and particularly preferably 1.
  • R 2 When there is a plurality of R 2 s, they may be the same or different.
  • R2 fluorine atom; chlorine atom; bromine atom; trifluoromethyl group; trichloromethyl group; tribromomethyl group; C1 to C4 alkyl group; C2 to C5 acyloxy group; C2 to C5 alkoxycarbonyl group; C1 to C3
  • An optionally substituted pyrrolidinylcarbonyl group is preferred, and includes a fluorine atom; a chlorine atom; a trifluoromethyl group; a trichloro
  • R 2 is one of the two carbon atoms located at the ortho position to the carbon atom directly bonded to the nitrogen atom forming the piperazine ring, among the carbon atoms forming the benzene ring to which R 2 is bonded. Or preferably, it is bonded to two carbon atoms, and more preferably to one ortho carbon atom. R 2 may be bonded to a carbon atom located at the para position. R 2 is (a) bonded to only one of the two carbon atoms in the ortho position, (b) one of the two carbon atoms in the ortho position and a carbon atom in the para position. and (c) preferably one of the two carbon atoms at the ortho position and one of the two carbon atoms at the meta position. R2 is (a) bonded to only one of the two carbon atoms in the ortho position, or (b) a carbon atom in the para position to one of the two ortho carbon atoms. More preferably, it is bonded to.
  • R2 bonded to the carbon atom located at the ortho position is preferably a halogen atom, a trihalogenomethyl group, or a C1-C3 alkyl group, such as a fluorine atom, a chlorine atom, a bromine atom, a trifluoromethyl group, a methyl group, or an ethyl group.
  • a group is more preferable, and a chlorine atom, a trifluoromethyl group, or a methyl group is particularly preferable.
  • R2 bonded to the carbon atom located at the para position is substituted with one or two groups selected from the group consisting of, for example, a halogen atom, an alkyl group; an acyloxy group; an alkoxycarbonyl group; an alkyl group and an alkoxycarbonyl group.
  • an amide group which may be substituted a piperidinylcarbonyl group which may be substituted with one or more groups selected from the group consisting of an alkyl group and an alkoxycarbonyl group; or a group consisting of an alkyl group and an alkoxycarbonyl group It may be a pyrrolidinylcarbonyl group which may be substituted with one or more groups selected from: chlorine atom; bromine atom; fluorine atom; C1-C3 alkyl group; C2-C4 acyloxy group; C2 -C4 alkoxycarbonyl group; amide group optionally substituted with one or two groups selected from the group consisting of C1-C3 alkyl group and C2-C4 alkoxycarbonyl group; C1-C3 alkyl group and C2-C4 a piperidinylcarbonyl group optionally substituted with one or more groups selected from the group consisting of alkoxycarbonyl groups; or selected from the group consisting of C1-C3
  • amide group substituted with one or two groups piperidinylcarbonyl group; or piperidinyl optionally substituted with one or more groups selected from methoxycarbonyl group and ethoxycarbonyl group; More preferred is a carbonyl group.
  • n is preferably an integer from 0 to 4, more preferably an integer from 0 to 3, even more preferably 0, 1, or 2, even more preferably 1 or 2, and particularly preferably 2.
  • the "halogen atom” includes a fluorine atom, a chlorine atom, a bromine atom, and an iodine atom.
  • Preferable examples include fluorine atom, chlorine atom, and bromine atom, more preferably fluorine atom and chlorine atom.
  • the "alkyl group” includes, for example, a C1-C8 alkyl group containing a linear, branched, or cyclic structure, preferably a C1-C6 alkyl group, more preferably a C1-C4 alkyl group. groups, particularly preferably C1-C3 alkyl groups.
  • linear or branched alkyl groups include methyl group, ethyl group, 1-propyl group, 2-propyl group, 1-butyl group, 2-butyl group, isobutyl group, and tert-butyl group.
  • alkyl groups containing a cyclic structure examples include cyclopropyl group, cyclopropylmethyl group, cyclobutyl group, cyclo Examples include butylmethyl group, cyclopentyl group, cyclopentylmethyl group, cyclohexyl group, cyclohexylmethyl group, cyclohexylethyl group, and the like.
  • Preferred examples include methyl group, ethyl group, 2-propyl group, t-butyl group, and cyclopropyl group.
  • the "alkyl group substituted with a halogen atom” includes, for example, the alkyl group substituted with a halogen atom and having a linear, branched, or cyclic structure.
  • the number of substituents an alkyl group has is 1 to the maximum number that can be substituted, preferably 3 to the maximum number that can be substituted.
  • there is a plurality of halogen atoms they may be the same or different, but are preferably the same.
  • Examples include C1 to C4 alkyl groups substituted with the maximum number of substitutable halogen atoms, preferably C1 to C3 alkyl groups substituted with the maximum number of substitutable halogen atoms, more preferably substitutable A C1-C2 alkyl group substituted with the maximum number of halogen atoms, particularly preferably a trihalogenomethyl group.
  • alkyl groups substituted with halogen atoms include perfluoro-n-butyl group, perfluoro-t-butyl group, perfluoro-n-propyl group, perfluoro-i-propyl group, pentylfluoroethyl group, Trifluoromethyl group, perchloro-n-butyl group, perchloro-t-butyl group, perchloro-n-propyl group, perchloro-i-propyl group, pentachloroethyl group, trichloromethyl group, perbromo-n-butyl group, perbromo -t-butyl group, perbromo-n-propyl group, perbromo-i-propyl group, pentabromoethyl group, tribromomethyl group, periodo-n-butyl group, periodo-t-butyl group, periodo-n-propyl group , periodo-i-propyl
  • Preferred examples of the alkyl group substituted with a halogen atom include a pentylfluoroethyl group, a trifluoromethyl group, a pentachloroethyl group, a trichloromethyl group, a pentabromoethyl group, a tribromomethyl group, a pentayodoethyl group, and a triiodoethyl group.
  • a methyl group is mentioned, a trifluoromethyl group, a trichloromethyl group, and a tribromomethyl group are more preferable, and a trifluoromethyl group and a trichloromethyl group are even more preferable.
  • the "alkoxy group” includes, for example, a C1-C8 alkoxy group containing a linear, branched, or cyclic structure, preferably a C1-C6 alkoxy group, more preferably a C1-C4 alkoxy group. groups, particularly preferably C1-C3 alkoxy groups.
  • linear or branched alkoxy groups include methoxy group, ethoxy group, 1-propoxy group, 2-propoxy group, 1-butoxy group, 2-butoxy group, isobutoxy group, and tert-butoxy group.
  • alkoxy groups containing a cyclic structure include cyclopropoxy group, cyclopropylmethoxy group, cyclobutyroxy group, cyclobutylmethoxy group, cyclopentyloxy group, cyclopentylmethoxy group, cyclohexyloxy group, cyclohexylmethoxy group, cyclohexylethoxy group, etc.
  • Preferred examples include methoxy group, ethoxy group, 2-propoxy group, tert-butoxy group, and cyclopropoxy group.
  • the "acyloxy group” includes, for example, a C2-C9 acyloxy group containing a linear, branched, or cyclic structure, preferably a C2-C7 acyloxy group, more preferably a C2-C5 acyloxy group. groups, particularly preferably C2-C4 acyloxy groups.
  • the linear or branched acyloxy group includes a methoxycarbonyl group, an ethoxycarbonyl group, a 1-propoxycarbonyl group, a 2-propoxycarbonyl group, a 1-butoxycarbonyl group, a 2-butoxycarbonyl group, Examples include isobutoxycarbonyl group, tert-butoxycarbonyl group, n-pentyloxycarbonyl group, neopentyloxycarbonyl group, n-hexyloxycarbonyl group, isohexyloxycarbonyl group, and 3-methylpentyloxycarbonyl group.
  • Acyloxy groups containing a cyclic structure include cyclopropoxycarbonyl group, cyclopropylmethoxycarbonyl group, cyclobutyroxycarbonyl group, cyclobutylmethoxycarbonyl group, cyclopentyloxycarbonyl group, cyclopentylmethoxycarbonyl group, cyclohexyloxycarbonyl group, cyclohexylmethoxy Examples include carbonyl group, cyclohexylethoxycarbonyl group, and the like. Preferred examples include methoxycarbonyl group, ethoxycarbonyl group, 2-propoxycarbonyl group, tert-butoxycarbonyl group, and cyclopropoxycarbonyl group.
  • the "alkoxycarbonyl group” includes a carbonyl group bonded to the alkoxy group, such as a carbonyl group bonded to a C1 to C8 alkoxy group, which has a linear, branched, or cyclic structure. and is preferably a C2-C7 alkoxycarbonyl group, more preferably a C2-C5 alkoxycarbonyl group, particularly preferably a C2-C4 alkoxycarbonyl group.
  • linear or branched alkoxy groups constituting the alkoxycarbonyl group include methoxy group, ethoxy group, 1-propoxy group, 2-propoxy group, 1-butoxy group, 2-butoxy group, isobutoxy group, tert- Examples include butoxy group, n-pentyloxy group, neopentyloxy group, n-hexyloxy group, isohexyloxy group, and 3-methylpentyloxy group.
  • alkoxy groups containing a cyclic structure constituting an alkoxycarbonyl group include a cyclopropoxy group, a cyclopropylmethoxy group, a cyclobutyroxy group, a cyclobutylmethoxy group, a cyclopentyloxy group, a cyclopentylmethoxy group, a cyclohexyloxy group, and a cyclohexylmethoxy group. , cyclohexylethoxy group, and the like.
  • Preferred examples of the alkoxy group constituting the alkoxycarbonyl group include methoxy group, ethoxy group, 2-propoxy group, tert-butoxy group, and cyclopropoxy group.
  • an amide group which may be substituted with one or two groups selected from the group consisting of an alkyl group and an alkoxycarbonyl group means an amide group, a mono- or di-alkyl amide group, a mono- or di- It includes an alkoxycarbonylamide group, an alkyl group, and an amide group di-substituted with an alkoxycarbonyl group, preferably an amide group or a dialkylamide group, and more preferably a dialkylamide group.
  • the substituents may be the same or different.
  • Examples of the monoalkyl amide group include amide groups mono-substituted with the above-mentioned alkyl groups, preferably amide groups mono-substituted with C1-C4 alkyl groups, more preferably amide groups mono-substituted with C1-C3 alkyl groups. Among these groups, particularly preferred are methylamide and ethylamide groups.
  • Examples of dialkylamide groups include amide groups di-substituted with the above-mentioned alkyl groups, preferably amide groups di-substituted with C1-C4 alkyl groups, more preferably amide groups di-substituted with C1-C3 alkyl groups.
  • dimethylamide group diethylamide group, and ethylmethylamide group.
  • the monoalkoxycarbonylamide group include an amide group mono-substituted with the above-mentioned alkoxycarbonyl group, preferably an amide group mono-substituted with a C2-C5 alkoxycarbonyl group, more preferably a mono-substituted amide group with a C2-C4 alkoxycarbonyl group.
  • Substituted amide groups particularly preferred are methoxycarbonylamide and ethoxycarbonylamide groups.
  • the dialkoxycarbonylamide group includes an amide group di-substituted with the alkoxycarbonyl group described above, preferably an amide group di-substituted with a C2-C5 alkoxycarbonyl group, more preferably a di-substituted amide group with a C2-C4 alkoxycarbonyl group.
  • Substituted amide groups particularly preferred are di(methoxycarbonyl)amide and di(ethoxycarbonyl)amide groups.
  • the amide group substituted with an alkyl group and an alkoxycarbonyl group includes an amide group di-substituted with the alkyl group and the alkoxycarbonyl group, preferably a di-substituted amide group with a C1-C4 alkyl group and a C2-C5 alkoxycarbonyl group.
  • a substituted amide group more preferably a di-substituted amide group with a C1 to C3 alkyl group and a C2 to C4 alkoxycarbonyl group, particularly preferably a methylmethoxycarbonylamide group, a methylethoxycarbonylamide group, an ethylmethoxycarbonylamide group, Examples include ethyl ethoxycarbonylamide group.
  • the number of substituents that the piperidinylcarbonyl group has is The number is 0 to 5, preferably 0 to 4, more preferably 0 to 3, even more preferably 0 to 2, particularly preferably 0 or 1.
  • the piperidinylcarbonyl group has a substituent at least one substituent is attached to the carbon atom at the 3-position of the piperidine ring (specifically, the 3-position when the nitrogen atom constituting the piperidine ring is the 1-position). Preferably, they are bonded.
  • substituents include the above-mentioned alkyl group and the above-mentioned alkoxycarbonyl group, preferably a C1-C4 alkyl group, a C2-C5 alkoxycarbonyl group, more preferably a C1-C3 alkyl group, a C2-C4 alkoxycarbonyl group. Particularly preferred are a methyl group, an ethyl group, a methoxycarbonyl group, and an ethoxycarbonyl group.
  • the number of substituents that the pyrrolidinylcarbonyl group has is The number is 0 to 4, preferably 0 to 3, more preferably 0 to 2, particularly preferably 0 or 1. When there are multiple substituents, they may be the same or different.
  • the pyrrolidinylcarbonyl group has a substituent at least one substituent is attached to the carbon atom at the 3-position of the pyrrolidine ring (specifically, the 3-position when the nitrogen atom constituting the pyrrolidine ring is the 1-position). Preferably, they are bonded.
  • substituents include the above-mentioned alkyl group and the above-mentioned alkoxycarbonyl group, preferably a C1-C4 alkyl group, a C2-C5 alkoxycarbonyl group, more preferably a C1-C3 alkyl group, a C2-C4 alkoxycarbonyl group. Particularly preferred are a methyl group, an ethyl group, a methoxycarbonyl group, and an ethoxycarbonyl group.
  • prevention means preventing the onset of heart failure or skeletal muscle failure in advance.
  • treatment means: (1) Delaying the onset of heart failure or skeletal muscle failure; (2) Slowing down or stopping the progression, exacerbation, or deterioration of symptoms of heart failure or skeletal muscle failure; (3) Heart failure or (4) means a method or process aimed at curing heart failure or skeletal muscle failure. Treatment may be administered before the onset of the disease or condition as a prophylactic measure, or alternatively, treatment can be administered after the onset of the disease.
  • heart failure refers to an abnormality in heart function in which the heart does not pump blood at the rate required by metabolic tissues.
  • Heart failure includes a wide range of disease states such as congestive heart failure, myocardial infarction, tachycardia, familial hypertrophic cardiomyopathy, ischemic heart disease, idiopathic dilated cardiomyopathy, and myocarditis.
  • Heart failure can result from many causes including, but not limited to, ischemic, congenital, rheumatic, viral, toxic or idiopathic forms.
  • Chronic cardiac hypertrophy a component of heart failure, is a serious disease state that is a precursor to congestive heart failure and cardiac arrest.
  • the present invention also encompasses pharmaceutical compositions for preventing and/or treating chronic cardiac hypertrophy.
  • skeletal muscle insufficiency refers to a state in which muscles do not have the ability to perform their work, and symptoms appear as a decrease in muscle strength, a decrease in endurance, or both. This includes muscular dystrophy.
  • a pharmaceutical composition usually means a drug for preventing or treating a disease, or for testing or diagnosing a disease.
  • the TRPC3/6/7 channel activator of the present invention does not lower blood pressure and exhibits efficacy against acute exacerbation.
  • TRPC3/6/7 channel inhibitors are not known to have any effect on acute exacerbation. Therefore, the present invention provides a treatment method in which the TRPC3/6/7 channel activator of the present invention is administered and then the TRPC3/6/7 channel inhibitor is administered in the early acute stage of heart failure or skeletal muscle failure. , is considered ideal.
  • the pharmaceutical composition of the present invention can be formulated by methods known to those skilled in the art.
  • the pharmaceutical composition of the present invention can be administered either parenterally or orally.
  • the composition can be, for example, in the form of an injection, a nasal administration, a pulmonary administration, or a transdermal administration.
  • parenteral administration the composition can be administered systemically or locally by intravenous injection, intramuscular injection, intraperitoneal injection, subcutaneous injection, etc.
  • the composition may be a tablet, capsule, pill, granule, powder, syrup, or the like.
  • the administration method can be selected as appropriate depending on the age and symptoms of the patient.
  • the dosage of the pharmaceutical composition containing the polypeptide can be set, for example, in the range of 0.0001 mg to 1000 mg per kg of body weight per dose. Alternatively, the dosage may be, for example, 0.001 to 100000 mg per patient, although the present invention is not necessarily limited to these values.
  • the dosage and method of administration vary depending on the patient's weight, age, symptoms, etc., and those skilled in the art can take these conditions into account and set an appropriate dosage and method of administration.
  • Another aspect of the present invention is a method for treating or preventing heart failure or skeletal muscle failure, which comprises administering a TRPC3/6/7 channel activator to a subject in need of such treatment. , preferably administering a compound of the invention to such a subject.
  • the present invention relates to TRPC3/6/7 channel activators for treating or preventing heart failure or skeletal muscle failure.
  • the present invention relates to the use of a TRPC3/6/7 channel activator for the manufacture of a medicament for treating or preventing heart failure or skeletal muscle failure.
  • formula (1) [In the formula, A represents a carbon atom or a nitrogen atom, B represents a carbon atom or a nitrogen atom, R 1 represents a halogen atom, an alkyl group, an alkoxy group, an acyloxy group, an alkoxycarbonyl group, or an alkyl group substituted with a halogen atom, and when there is more than one, they may be the same or different; m represents an integer from 0 to 5, n represents an integer from 2 to 5, At least one of the plurality of R 2 is a halogen atom, an alkyl group, or a trifluoromethyl group; the remaining R 2 are 1) substituted with one or more halogen atoms, and are selected from an alkyl group and an alkoxycarbonyl group.
  • the present invention provides a method of screening for substances for preventing and/or treating heart failure or skeletal muscle failure, the screening method selecting for TRPC3/6/7 channel activators.
  • the screening method of the present invention includes a step of measuring zinc concentration in cells in the presence of a test substance, and a step in which the zinc concentration increases compared to the zinc concentration in the absence of the test substance.
  • a method for screening a candidate substance for a TRPC3/6/7 channel activating substance comprising the step of determining that the test substance is a candidate substance that causes activation of a TRPC3/6/7 channel.
  • the cells are not limited as long as they are cultured cells. For example, when using cultured mouse or rat cardiomyocytes, it is preferable to isolate them from the heart and perform primary culture.
  • HEK293 cells are used as a commercially available cell line, and activity can be observed by overexpressing TRPC3/6/7 channels.
  • the cells are preferably HEK293 cells, more preferably cultured mouse or rat cardiomyocytes.
  • test substance is a substance that can be evaluated to determine whether it is a candidate substance for a TRPC3/6/7 channel activator, and is not particularly limited. Examples include compounds, proteins, peptides, nucleic acids, lipids, carbohydrates, glycolipids, glycoproteins, metals, and the like. The method of administering the test substance is also not particularly limited.
  • Reference example 1 Compensatory cardiac hyperactivity in response to a transient decrease in blood pressure due to intravenous administration of the vasodilator hydralazine (Hyd) Wild type (WT) and TRPC6 gene-deficient mice (TRPC6(-/-) obtained from the NIEHS Institute in the United States) (Dietrich A, Mederos YSM, Gollasch M, Gross V, Storch U, Dubrovska G, et al. (2005). Increased vascular smooth muscle contractility in TRPC6-/- mice. Molecular and cellular biology 25(16): 6980-6989.
  • FIG. 1 shows that compared to wild type (WT) mice, the increase in left ventricular contractile force ( ⁇ LVdP/dtmax) is significantly attenuated in TRPC6 gene-deficient mice, while there is no difference in the increase in heart rate ( ⁇ Heart Rate). It is shown that.
  • TRPC6-deficient mice showed a positive change in hydralazine-induced baroreflex responses (positive chronotropy (increase in heart rate) and positive inotropy (increase in contractile force)) compared to wild-type mice. This means that the force action is significantly weakened.
  • TRPC3/6/7 channel activator PPZ2, perforin, and GSK1702934A used in the test example are represented by the following formula.
  • Test example 1 TRPC6 channel activation in MLP-deficient mice
  • Muscle LIM protein (MLP)-deficient mouse a dilated cardiomyopathy model mouse (Knoll R, Hoshijima M, Hoffman HM, Person V, Lorenzen-Schmidt I, Bang ML, Hayashi T, Shiga N, Yasukawa H, Schaper W, McKenna W, Yokoyama M, Schork NJ, Omens JH, McCulloch AD, Kimura A, Gregorio CC, Poller W, Schaper J, Schultheiss HP and Chien KR
  • the cardiac mechanical stretch sensor machinery involves a Z disc complex that is defective in a sub set of human dilated cardiomyopathy. Cell 111 (2002b), pp. 943-55.) develops heart failure 5 weeks after birth.
  • Test example 2 TRPC6 channel activation in pressure overload-induced heart failure model mice
  • Pressure overload-induced heart failure model mice Male mice of the 129Sv strain (Kitajima N, et al. TRPC3 positively regulates reactive oxygen species driving maladaptive cardiac remodeling. Sci Rep 6, 37001 (2016))
  • TAC transverse aortic constriction
  • PPZ2 phosphodiesterase 5A regulates cardiac adrenergic stimulation by NOS3-dependent mechanism.
  • Figure 3-2f TRPC6 deficiency
  • Fig. 3 shows that PPZ2-treated mice had higher levels of cancer compared to mice treated with vehicle (50% (v/v) diethyl sulfoxide (DMSO)/50% (v/v) polyethylene glycol 300) (control). , showing a significant improvement in heart failure after TAC.
  • vehicle 50% (v/v) diethyl sulfoxide (DMSO)/50% (v/v) polyethylene glycol 300
  • Test example 3 TRPC6 channel activation in post-myocardial infarction heart failure model mice (Nishimura A, et al. Hypoxia-induced interaction of filamin with Drp1 causes mitochondrial hyperfission-associated myocardial senescence. Sci Signal 11, (2016).)
  • the left anterior descending coronary artery was ligated to induce myocardial infarction (MI).
  • MI myocardial infarction
  • an osmotic pump containing PPZ2 was implanted, and subsequent changes in cardiac function were analyzed.
  • MI cardiac function gradually decreased over time in control mice, whereas in PPZ-treated mice, the rate of increase in EF and FS increased significantly for 3 to 4 weeks after administration ( Figure 4 ).
  • Test example 4 Effect on baroreceptive reflex response by inducing a transient decrease in blood pressure Hydralazine (0.5 mg/kg, 1.2 mL/h) was administered to mice that had been intraperitoneally administered PPZ2 30 minutes prior to inducing a transient decrease in blood pressure.
  • TRPC6 channel inhibitor BI749327 (30 mg/kg) (In vivo selective inhibition of TRPC6 by antagonist BI 749327 ameliorates fibrosis and dysfunction in cardiac and renal disease, https://doi.org/10.1073/pnas.1815354116) was combined with PPZ2 ( 2.5 mg/kg) was administered orally 5 hours before administration.
  • the obtained results are shown in Figure 5.
  • the vertical axis shows the hydralazine-induced increase in left ventricular contractile force ( ⁇ LVdP/dtmax) or heart rate ( ⁇ Heart Rate).
  • Administration of PPZ2 increases hydralazine-induced ⁇ LVdP/dtmax in a PPZ2 concentration-dependent manner.
  • PPZ2 administration did not change the hydralazine-induced ⁇ Heart Rate.
  • the hydralazine-induced increase in ⁇ LVdP/dtmax due to administration of PPZ2 was suppressed by administration of BI749327, a TRPC6 channel inhibitor. From the above, it was found that PPZ2 enhances the positive inotropic effect of the heart caused by the baroreceptive reflex in a concentration-dependent manner through activation of the TRPC6 channel.
  • Test example 5 Zn 2+ concentration enhancement effect of TRPC6 channel activator GSK1702934A TRPC6 channel activator other than PPZ2 GSK1702934A (In vivo selective inhibition of TRPC6 by antagonist BI 749327 ameliorates fibrosis and dysfunction in cardiac and renal disease, https://doi.org /10.1073/pnas.1815354116) was investigated to enhance Zn 2+ concentration.
  • TRPC6 WT
  • Zn 2+ permeability-defective mutant KYD
  • Test example 6 Effect of TRPC6 channel activator GSK1702934A and hyperforin on enhancing myocardial contractility TRPC6 channel activator other than PPZ2 GSK1702934A and hyperforin (GSK1702934A and M085 directly activate TRPC6 via a mechanism of stimulating the extracellular cavity formed by the pore helix and transmembrane helix S6, https://doi.org/10.1016/j.jbc.2021.101125) to investigate the effect of enhancing myocardial contractility.
  • Neonatal rat cardiomyocytes were treated with a TRPC6 activator for 30 minutes to examine whether norepinephrine (NE) (1 ⁇ M) increased the contractile force of cardiomyocytes.
  • NE norepinephrine
  • FIG. PPZ2 enhanced NE-induced myocardial contraction.
  • the myocardial contraction enhancing effect of PPZ2 was attenuated by treatment with TPEN, a Zn 2+ chelator. Therefore, it was revealed that PPZ2 enhances NE-induced myocardial contraction in a Zn 2+ -dependent manner (Fig. 7a). It was found that GSK1702934A and hyperforin also showed effects similar to PPZ2 ( Figure 7b).
  • Test example 7 TRPC6 channel activation in muscular dystrophy model mice
  • Muscular dystrophy model mice C57BL/10-mdx mice (6 weeks old, purchased from CLEA Japan) were implanted with an osmotic pump containing PPZ2 (2.5 mg/kg) intraperitoneally for 4 weeks. Continuous administration was performed.
  • PPZ2 blood levels, which are markers of myopathy, were significantly reduced compared to mice treated with vehicle (50% (v/v) diethyl sulfoxide (DMSO)/50% (v/v) polyethylene glycol 300) (control). It was found that LDH (lactate dehydrogenase) and CK (creatinine kinase) were significantly inhibited (Figure 8).
  • Preparation example 7 (3-chloro-4-(piperazin-1-yl)phenyl)(4,4-difluoropiperidin-1-yl)methanone dihydrochloride 1 of 4-(2-chloro-4-(4,4-difluoropiperidine-1-carbonyl)phenyl)piperazine-1-carboxylic acid tert-butyl ester (1933 mg, 4.35 mmol) prepared in Preparation Example 6 A 4 N HCl/1,4-dioxane solution (10 mL) was added to a solution of ,4-dioxane (5 mL), and the mixture was stirred at room temperature overnight.
  • Example 1 2-(4-(2-chloro-4-(4,4-difluoropiperidine-1-carbonyl)phenyl)piperazin-1-yl)-N-(2-ethoxyphenyl)acetamide (3-chloro-4-(piperazin-1-yl)phenyl)(4,4-difluoropiperidin-1-yl)methanone dihydrochloride (55 mg, 0.13 mmol) prepared in Preparation Example 7, Preparation Example 1 A solution of 2-bromo-N-(2-ethoxyphenyl)acetamide (52 mg, 0.2 mmol) prepared in The mixture was heated and stirred for 10 hours.
  • Example 2 2-(4-(2-chloro-4-(4,4-difluoropiperidine-1-carbonyl)phenyl)piperazin-1-yl)-N-(2-(trifluoromethoxy)phenyl)acetamide (3-chloro-4-(piperazin-1-yl)phenyl)(4,4-difluoropiperidin-1-yl)methanone dihydrochloride (41.7 mg, 0.1 mmol) prepared in Preparation Example 7, Preparation Example 2 A solution of 2-bromo-N-(2-(trifluoromethoxy)phenyl)acetamide (29.8 mg, 0.1 mmol) and DBU (60 ⁇ L, 0.4 mmol) in DMF (1 mL) was added under a nitrogen atmosphere.
  • Example 3 2-(4-(2-chloro-4-(4,4-difluoropiperidine-1-carbonyl)phenyl)piperazin-1-yl)-N-(2-(2,2,2-trifluoroethoxy) phenyl)acetamide (3-chloro-4-(piperazin-1-yl)phenyl)(4,4-difluoropiperidin-1-yl)methanone dihydrochloride (41.7 mg, 0.1 mmol) prepared in Preparation Example 7, Preparation Example 3 2-bromo-N-(2-(2,2,2-trifluoroethoxy)phenyl)acetamide (31.2 mg, 0.1 mmol) and DBU (54 ⁇ L, 0.4 mmol) in DMF (1 mL).
  • the solution was heated and stirred at 80° C. for 10 hours under a nitrogen atmosphere.
  • the reaction solution was returned to room temperature, a saturated aqueous ammonium chloride solution was added, and extraction was performed with ethyl acetate.
  • the organic layer was washed with a saturated aqueous sodium bicarbonate solution and then with saturated brine, dried over sodium sulfate, and concentrated.
  • the residue was purified by medium pressure preparative silica gel column chromatography (hexane/ethyl acetate) to obtain crystals of the desired product (14.8 mg). Yield 26%.
  • Test example 8 Influx of Ca 2+ into HEK293 cells that transiently express TRPC channels
  • TRPC3 or TRPC6 genes were introduced into HEK293 cells (human embryonic kidney-derived cells) by lipofection. TRPC6 channel was expressed in the cell membrane. That is, by introducing TRPC6 plasmid DNA (pCI-neo (Promega)) into HEK293 cells using a lipofection agent and culturing them at 37°C for 24 to 48 hours, HEK293 cells with TRPC6 channels expressed in their cell membranes were produced. (TRPC6-expressing HEK293 cells) were generated.
  • Non-Patent Document 1 S. Sawamura et al., Mol. Pharmacol., 89, 348-363 (2016)
  • TRPC3 HEK293 cells in which the TRPC3 channel was expressed in the cell membrane (TRPC3-expressing HEK293 cells) were produced in the same manner.
  • Ca2 + -containing solution (2mM CaCl2 , 132mM NaCl, 4mM KCl, 1mM MgCl2, 5mM glucose.5mM HEPES (pH7.4)
  • Ca2 + -free solution (132mM NaCl, 4mM KCl , 1mM MgCl2 , 5mM glucose, 5mM HEPES (pH7.4)).
  • Test example 8-1 Effect of Example Compounds on Ca 2+ Influx into HEK293 Cells Expressing TRPC6 Channel Changes in calcium ion concentration of the channel were measured according to the measurement method described in Test Example 8.
  • PPZ2, L687, L-0042 to L-0046 (compounds shown below) are used as test compounds, and the amount added is 1 to 30 ⁇ M. Additionally, 0.1% DMSO was used instead of the test compound as a control.
  • the fluorescence intensity (Ratio) of Fura-2 By measuring the fluorescence intensity (Ratio) of Fura-2 over time, the increase in Ca 2+ in HEK293 cells expressing TRPC6 channel (from the maximum Ratio value measured after addition of the test compound to the initial value (before addition of the test compound)) was determined. ⁇ Ratio value) was calculated.
  • Test compounds that caused a greater Ca 2+ increase than the DMSO control were designated as substances that activate TRPC6 channels.
  • L687 and PPZ2 are shown in FIG. 9, and the results for the compounds of the invention are shown in FIG. 10.
  • PPZ2 is known to activate TRPC6 channels and cause Ca 2+ to flow into cells, and in this study, addition of PPZ2 resulted in a concentration-dependent increase in the ⁇ Ratio value.
  • Similar compounds L687 ( Figure 9) and L-0042 to L-0046 ( Figure 10) also increased intracellular Ca 2+ concentration in a concentration-dependent manner, indicating that these compounds activate TRPC6 channels. was confirmed. In particular, the strongest TRPC6-dependent increase in intracellular Ca 2+ concentration was observed in L-0044 (Example 1 compound).
  • Test example 8-2 Effect of example compounds on Ca 2+ influx into HEK293 cells expressing TRPC3 channels TRPC3-expressing HEK293 cells (TRPC3/HEK293) prepared by the method described in Test Example 81 were used, and L-0042 was used as the test compound. ⁇ L-0046 (1-30 ⁇ M) was used. The amount of Ca 2+ increase ( ⁇ Ratio) into each cell was determined in the same manner as Test Example 8-1. The results obtained are shown in FIG. In HEK293 cells expressing human TRPC3 channels, compounds L-0042 to L-0046 of the present invention increased intracellular Ca 2+ concentration in a concentration-dependent manner, indicating that these compounds also activate TRPC3 channels. This was confirmed. In particular, the strongest TRPC3-dependent increase in intracellular Ca 2+ concentration was observed in L-0043 (Example 2 compound).
  • Test example 9 Suppressive effect of L-0044 on in vitro cardiac hypertrophic response
  • Neonatal rat cardiomyocytes were preincubated with or without 3 ⁇ M L-0044 (Example 1 compound) for 1 hour.
  • Cells were then stimulated with 100 nM endothelin-1 (ET-1) for 24 hours.
  • ET-1 endothelin-1
  • FIG. 12 shows that ET-1 administration induces cardiomyocyte hypertrophy
  • L-0044 pretreatment reduces ET-1-mediated cardiac hypertrophy. This revealed that although L-0044 alone tends to cause weak myocardial hypertrophy, treatment with 3 ⁇ M L-0044 suppresses pathological cardiac hypertrophy caused by endothelin-1 stimulation.
  • Test example 10 Effect of Example Compounds on Zn 2+ Influx into HEK293 Cells Expressing TRPC6 Channel Using TRPC6-expressing HEK293 cells (HEK293 expressing TRPC6-mCherry) or control HEK293 cells, L-0043, L- 0044, L-0046 (10 ⁇ M) or PPZ2 (30 ⁇ M) were used. In the same manner as Test Example 5, the amount of increase in Zn 2+ into each cell was determined. The obtained results are shown in FIG. 13. Figure 13 shows that L-0043 and L-0044 potently enhance Zn 2+ influx through TRPC6 at lower concentrations than PPZ2.
  • Test example 11 Neonatal rat cardiomyocytes were preincubated with L-0044 (1 ⁇ M) for 30 min. The cells were placed in a microscope equipped with a high-speed camera (60 fps) and videos of cell movement were taken. Cells were electrically stimulated for 3 minutes (5 V, 1 Hz, pulse width 10 ms) and then stimulated with norepinephrine (NE) (0.1 ⁇ M) to evaluate changes in cell movement. Cardiomyocyte contractility was analyzed using the ImageJ plug-in MUSCLEMOTION (A Versatile Open Software Tool to Quantify Cardiomyocyte and Cardiac Muscle Contraction In Vitro and In Vivo, https://doi.org/10.1161/CIRCRESAHA.117.312067). The obtained results are shown in FIG. 14. Figure 14 shows that L-0044 pretreatment enhances the contractile force of cardiac myocytes induced by NE stimulation.

Landscapes

  • Health & Medical Sciences (AREA)
  • Chemical & Material Sciences (AREA)
  • Life Sciences & Earth Sciences (AREA)
  • Medicinal Chemistry (AREA)
  • General Health & Medical Sciences (AREA)
  • Engineering & Computer Science (AREA)
  • Pharmacology & Pharmacy (AREA)
  • Animal Behavior & Ethology (AREA)
  • Public Health (AREA)
  • Veterinary Medicine (AREA)
  • Organic Chemistry (AREA)
  • Epidemiology (AREA)
  • Bioinformatics & Cheminformatics (AREA)
  • Immunology (AREA)
  • Molecular Biology (AREA)
  • Nuclear Medicine, Radiotherapy & Molecular Imaging (AREA)
  • General Physics & Mathematics (AREA)
  • Chemical Kinetics & Catalysis (AREA)
  • Urology & Nephrology (AREA)
  • Hematology (AREA)
  • Biochemistry (AREA)
  • General Chemical & Material Sciences (AREA)
  • Biomedical Technology (AREA)
  • Physics & Mathematics (AREA)
  • Pathology (AREA)
  • Analytical Chemistry (AREA)
  • Cardiology (AREA)
  • Food Science & Technology (AREA)
  • Physical Education & Sports Medicine (AREA)
  • Pharmaceuticals Containing Other Organic And Inorganic Compounds (AREA)
  • Orthopedic Medicine & Surgery (AREA)
  • Biophysics (AREA)
  • Biotechnology (AREA)
  • Cell Biology (AREA)
  • Neurology (AREA)
  • Microbiology (AREA)
  • Hospice & Palliative Care (AREA)
  • Heart & Thoracic Surgery (AREA)

Abstract

Provided is a drug for preventing or treating heart failure, that is effective and that does not have a blood pressure lowering effect.　The present invention pertains to: a pharmaceutical composition for preventing or treating heart failure or skeletal muscle failure, the pharmaceutical composition containing a TRPC3/6/7 channel activator; and a related screening method.

Description

ＴＲＰＣ３／６／７チャネル活性化による強心作用を介した心不全治療Heart failure treatment via inotropic effect by TRPC3/6/7 channel activation
　本特許出願は、日本国特許出願２０２２－１３２５３３号（２０２２年８月２３日出願）に基づくパリ条約上の優先権および利益を主張するものであり、ここに引用することによって、上記出願に記載された内容の全体が本明細書中に組み込まれるものとする。 This patent application claims priority and benefit under the Paris Convention based on Japanese Patent Application No. 2022-132533 (filed on August 23, 2022), and is hereby incorporated by reference. The entire contents thereof are incorporated herein by reference.
　本発明は一般に、TRPC3/6/7チャネル活性化物質の新たな用途に関する。本発明は詳細には、TRPC3/6/7チャネル活性化物質を含有する医薬組成物、具体的には心不全または骨格筋不全を予防および／または処置するための医薬組成物、および心不全または骨格筋不全を予防および／または処置するための物質をスクリーニングする方法に関する。 The present invention generally relates to new uses for TRPC3/6/7 channel activators. The present invention particularly relates to pharmaceutical compositions containing TRPC3/6/7 channel activators, specifically pharmaceutical compositions for preventing and/or treating heart failure or skeletal muscle failure, and The present invention relates to a method for screening substances for preventing and/or treating disorders.
　近年、社会の高齢化に伴って心不全が世界中で増加しており、「心不全パンデミック」と呼ぶべき事態が進行している。現在、世界の心不全患者数は3,000万人に達する勢いであり、その数は今後ますます増加し、病床を逼迫する可能性が懸念されている。心不全は一度発症すると、完治することなく急性増悪（急性心不全）と緩解を繰り返しながら徐々に心機能を低下し（慢性心不全）、やがて治療抵抗性心不全となり、死に至るという予後不良な病態である。心不全における治療に関しては、急性心不全と慢性心不全ではその方針、治療法がしばしば異なることがよく知られている。慢性心不全に対しては神経体液性因子の過剰な活性化が心負荷につながっているとの考えから心負荷を軽減する、神経体液性因子の遮断薬がその病態悪化やそれにより死亡を抑制することが示されている。一方、急性心不全ではまずは救命を優先し、血行動態の適正化を図ることが優先されるが、しばしば慢性心不全の治療方針とは相反する治療法を行わざるを得ないことも多く、現在までに急性心不全の治療薬として心不全の慢性期予後を改善する薬剤は存在せず、新規作用機序に基づく急性心不全治療薬が求められている。 In recent years, heart failure has been increasing around the world as society ages, and a situation that can be called a "heart failure pandemic" is progressing. Currently, the number of heart failure patients worldwide is on track to reach 30 million, and there are concerns that this number will continue to increase, leading to a shortage of hospital beds. Once developed, heart failure is a pathological condition with a poor prognosis, with no complete recovery, and a gradual decline in heart function (chronic heart failure) through repeated periods of acute exacerbation (acute heart failure) and remission (chronic heart failure), eventually leading to treatment-resistant heart failure and death. Regarding the treatment of heart failure, it is well known that the policies and treatments for acute heart failure and chronic heart failure often differ. In chronic heart failure, it is believed that excessive activation of neurohumoral factors is linked to cardiac stress, so neurohumoral factor blockers, which reduce cardiac stress, can suppress the deterioration of the condition and thereby death. It has been shown that On the other hand, in the case of acute heart failure, the first priority is to save life and optimize hemodynamics, but it is often necessary to perform treatment methods that conflict with the treatment policy for chronic heart failure. As a therapeutic agent for acute heart failure, there is no drug that improves the prognosis of the chronic phase of heart failure, and there is a need for a therapeutic agent for acute heart failure based on a new mechanism of action.
　心臓のポンプ活動や形態構造は、様々な神経体液性因子によって調節されている。心臓を構成する細胞は、これら神経体液性因子を細胞膜上で感知し、細胞内に適切なシグナルを伝達させることで心機能恒常性を維持している。心不全の発症・進展に伴って起こる心筋細胞の肥大や間質の線維化には、アンジオテンシンIIやエンドセリン１などホスホリパーゼCと連関した受容体の活性化を介するCa2+シグナル伝達、中でもCa2+依存性肥大関連遺伝子の発現誘導に関わる細胞外から細胞内へのCa2+流入が深く関わっており、受容体作動性のCa2+流入を担う分子実体として注目されているのがトランジエント・レセプター・ポテンシャル・カノニカル（transient receptor potential canonical：TRPC）タンパク質である。動物TRPCチャネルには7つのアイソフォーム（TRPC1-TRPC7）が存在し、中でも脂質（ジアシルグリセロール）作動性のTRPC3とTRPC6チャネルが心臓リモデリング形成に重要であることが、本出願人の成果も含めて、数多く報告されてきた。実際に、TRPC3/C6チャネル活性を阻害する化合物が病的心肥大や心不全を改善することも動物レベルで実証されており、治療標的としての有効性が示唆されている（非特許文献１－７）。また、特許文献１には、TRPC3チャネルまたはTRPC6チャネルの活性を調節または阻害するベンゾイソオキサゾール化合物が記載され、TRPC3および／またはTRPC6関連疾患として心肥大が挙げられているが、TRPC3/6/7チャネルを活性化する物質については記載されていない。いずれにせよ、心機能調節におけるTRPC3/6チャネルの生理的役割についてはよく理解できていない状況である。 The pumping activity and morphological structure of the heart are regulated by various neurohumoral factors. The cells that make up the heart maintain cardiac homeostasis by sensing these neurohumoral factors on their cell membranes and transmitting appropriate signals within the cells. Cardiomyocyte hypertrophy and interstitial fibrosis that occur with the onset and progression of heart failure are caused by Ca 2+ signaling, especially Ca 2+ The influx of Ca 2+ from outside the cell into the cell is deeply involved in the induction of expression of genes related to hypertrophy, and transient Ca 2+ influx is attracting attention as a molecular entity responsible for receptor-mediated Ca 2+ influx. It is a receptor potential canonical (TRPC) protein. There are seven isoforms (TRPC1-TRPC7) of animal TRPC channels, and among them, lipid (diacylglycerol)-operated TRPC3 and TRPC6 channels are important for cardiac remodeling, including the results of the present applicant. There have been many reports. In fact, it has been demonstrated at the animal level that compounds that inhibit TRPC3/C6 channel activity improve pathological cardiac hypertrophy and heart failure, suggesting their effectiveness as therapeutic targets (Non-patent Documents 1-7 ). Further, Patent Document 1 describes a benzisoxazole compound that modulates or inhibits the activity of TRPC3 channel or TRPC6 channel, and mentions cardiac hypertrophy as a TRPC3 and/or TRPC6-related disease. Substances that activate the channels are not described. In any case, the physiological role of TRPC3/6 channels in regulating cardiac function is not well understood.
WO2019208812公報WO2019208812 publication
　上記の通り、予後不良な病態である心不全を治療する治療薬は長年にわたり、臨床上、処方されているが、その多くは降圧作用を持つことや、急性心不全や慢性心不全の急性増悪に対する有効な治療をもたらさない。このことから、より有効な強心薬の開発が求められている。 As mentioned above, therapeutic drugs to treat heart failure, a condition with a poor prognosis, have been clinically prescribed for many years, but many of them have antihypertensive effects and are effective against acute heart failure and acute exacerbation of chronic heart failure. Does not provide treatment. For this reason, there is a need for the development of more effective cardiotonic drugs.
　本発明者らは、TRPC6遺伝子欠損マウスが心不全に対して脆弱な傾向を示すこと、すなわちTRPC6欠損マウスの心臓が高血圧負荷に適応できていないことに注目し、TRPC3/6/7チャネルを活性化する物質の病態への応用を試みた。種々の動物実験により、異なる心不全モデル、異なるマウス系統種であっても、TRPC3/6/7チャネルを活性化する物質が同じように心不全保護効果があることを見出し、本発明を完成させた。 The present inventors focused on the tendency of TRPC6 gene-deficient mice to be vulnerable to heart failure, that is, the heart of TRPC6-deficient mice was unable to adapt to high blood pressure load, and activated TRPC3/6/7 channels. We attempted to apply substances that cause disease to pathological conditions. Through various animal experiments, we discovered that substances that activate TRPC3/6/7 channels have the same protective effect on heart failure even in different heart failure models and different mouse strains, and have completed the present invention.
　したがって、本発明は、以下の態様を含む。
＜医薬組成物＞
［１］
　TRPC3/6/7チャネル活性化物質を含有する、心不全または骨格筋不全を予防および／または処置するための医薬組成物。
［２］
　TRPC3/6/7チャネル活性化物質が、式（１）：
Figure JPOXMLDOC01-appb-C000007
 ［式中、
　Ａは、炭素原子または窒素原子を示し、
　Ｂは、炭素原子または窒素原子を示し、
　Ｒ１は、置換されてもよいアルキル基を示し、複数あるときは同一または異なってよく、
　ｍは、０～５の整数を示し、
　Ｒ２は、ハロゲン原子；トリハロゲノメチル基；アルキル基；アシルオキシ基；アルコキシカルボニル基；置換されてもよいアミド基；置換されてもよいピペリジニルカルボニル基；或いは置換されてもよいピロリジニルカルボニル基を示し、複数あるときは同一または異なってよく、
　ｎは、０～５の整数を示し、
　ｐは、０または１を示し、そして
　ｑは、０または１を示す。］
で表される化合物、ハイパーフォリン、GSK1702934A、およびカンナビジオール；
からなる群から選択される少なくとも１種の化合物である、［１］記載の医薬組成物。 Accordingly, the present invention includes the following aspects.
<Pharmaceutical composition>
[1]
A pharmaceutical composition for preventing and/or treating heart failure or skeletal muscle failure, containing a TRPC3/6/7 channel activator.
[2]
The TRPC3/6/7 channel activator has the formula (1):
Figure JPOXMLDOC01-appb-C000007
 [In the formula,
A represents a carbon atom or a nitrogen atom,
B represents a carbon atom or a nitrogen atom,
R 1 represents an optionally substituted alkyl group, and when there is a plurality of them, they may be the same or different,
m represents an integer from 0 to 5,
R 2 is a halogen atom; a trihalogenomethyl group; an alkyl group; an acyloxy group; an alkoxycarbonyl group; an optionally substituted amide group; an optionally substituted piperidinylcarbonyl group; or an optionally substituted pyrrolidinyl Indicates a carbonyl group, and when there are multiple groups, they may be the same or different,
n represents an integer from 0 to 5,
p represents 0 or 1, and q represents 0 or 1. ]
Compounds represented by, hyperforin, GSK1702934A, and cannabidiol;
The pharmaceutical composition according to [1], which is at least one compound selected from the group consisting of.
［３］
　TRPC3/6/7チャネル活性化物質が、式（１）：
Figure JPOXMLDOC01-appb-C000008
 ［式中、
　Ａは、炭素原子または窒素原子を示し、
　Ｂは、炭素原子または窒素原子を示し、
　Ｒ１は、ハロゲン原子、アルキル基、アルコキシ基、アシルオキシ基、アルコキシカルボニル基、またはハロゲン原子で置換されたアルキル基を示し、複数あるときは同一または異なってよく、
　ｍは、０～５の整数を示し、
　Ｒ２は、ハロゲン原子；トリハロゲノメチル基；アルキル基；アシルオキシ基；アルコキシカルボニル基；アルキル基、アルコキシカルボニル基およびハロゲン原子からなる群から選択される１種または２種の基で置換されてもよいアミド基；アルキル基、アルコキシカルボニル基およびハロゲン原子からなる群から選択される１種または２種以上の基で置換されてもよいピペリジニルカルボニル基；或いはアルキル基、アルコキシカルボニル基およびハロゲン原子からなる群から選択される１種または２種以上の基で置換されてもよいピロリジニルカルボニル基を示し、複数あるときは同一または異なってよく、
　ｎは、０～５の整数を示し、
　ｐは、０または１を示し、そして
　ｑは、０または１を示す。］
で表される化合物、ハイパーフォリン、GSK1702934A、およびカンナビジオール；
からなる群から選択される少なくとも１種の化合物である、［２］記載の医薬組成物。
［４］
　前記TRPC3/6/7チャネル活性化物質が、前記式（１）で表わされる化合物であって、
　Ａは、炭素原子または窒素原子を示し、
　Ｂは、炭素原子または窒素原子を示し、
　Ｒ１は、フッ素原子、塩素原子、臭素原子、Ｃ１～Ｃ４アルキル基、Ｃ１～Ｃ４アルコキシ基、Ｃ２～Ｃ５アシルオキシ基、Ｃ２～Ｃ５アルコキシカルボニル基、またはトリハロゲノメチル基を示し、複数あるときは同一または異なってよく、
　ｍは、０～４の整数を示し、
　Ｒ２は、フッ素原子；塩素原子；臭素原子；トリフルオロメチル基；トリクロロメチル基；トリブロモメチル基；Ｃ１～Ｃ４アルキル基；Ｃ２～Ｃ５アシルオキシ基；Ｃ２～Ｃ５アルコキシカルボニル基；Ｃ１～Ｃ３アルキル基およびＣ２～Ｃ４アルコキシカルボニル基からなる群から選択される１種または２種の基で置換されてもよいアミド基；Ｃ１～Ｃ３アルキル基、Ｃ２～Ｃ４アルコキシカルボニル基およびハロゲン原子からなる群から選択される１種または２種以上の基で置換されてもよいピペリジニルカルボニル基；或いはＣ１～Ｃ３アルキル基、Ｃ２～Ｃ４アルコキシカルボニル基、およびハロゲン原子からなる群から選択される１種または２種以上の基で置換されてもよいピロリジニルカルボニル基を示し、複数あるときは同一または異なってよく、
　ｎは、０～４の整数を示し、
　ｐは、０または１を示し、そして
　ｑは、０または１を示す、
［３］に記載の医薬組成物。 [3]
The TRPC3/6/7 channel activator has the formula (1):
Figure JPOXMLDOC01-appb-C000008
 [In the formula,
A represents a carbon atom or a nitrogen atom,
B represents a carbon atom or a nitrogen atom,
R 1 represents a halogen atom, an alkyl group, an alkoxy group, an acyloxy group, an alkoxycarbonyl group, or an alkyl group substituted with a halogen atom, and when there is more than one, they may be the same or different;
m represents an integer from 0 to 5,
R 2 may be substituted with one or two groups selected from the group consisting of a halogen atom; a trihalogenomethyl group; an alkyl group; an acyloxy group; an alkoxycarbonyl group; an alkyl group, an alkoxycarbonyl group, and a halogen atom. A good amide group; a piperidinylcarbonyl group which may be substituted with one or more groups selected from the group consisting of an alkyl group, an alkoxycarbonyl group, and a halogen atom; or an alkyl group, an alkoxycarbonyl group, and a halogen atom Indicates a pyrrolidinylcarbonyl group which may be substituted with one or more groups selected from the group consisting of, and when there are multiple groups, they may be the same or different,
n represents an integer from 0 to 5,
p represents 0 or 1, and q represents 0 or 1. ]
Compounds represented by, hyperforin, GSK1702934A, and cannabidiol;
The pharmaceutical composition according to [2], which is at least one compound selected from the group consisting of.
[4]
The TRPC3/6/7 channel activator is a compound represented by the formula (1),
A represents a carbon atom or a nitrogen atom,
B represents a carbon atom or a nitrogen atom,
R 1 represents a fluorine atom, a chlorine atom, a bromine atom, a C1-C4 alkyl group, a C1-C4 alkoxy group, a C2-C5 acyloxy group, a C2-C5 alkoxycarbonyl group, or a trihalogenomethyl group, and when there is more than one, may be the same or different;
m represents an integer from 0 to 4,
R 2 is a fluorine atom; a chlorine atom; a bromine atom; a trifluoromethyl group; a trichloromethyl group; a tribromomethyl group; a C1 to C4 alkyl group; a C2 to C5 acyloxy group; a C2 to C5 alkoxycarbonyl group; a C1 to C3 alkyl and an amide group which may be substituted with one or two groups selected from the group consisting of a C2-C4 alkoxycarbonyl group; a C1-C3 alkyl group, a C2-C4 alkoxycarbonyl group, and a halogen atom; a piperidinylcarbonyl group which may be substituted with one or more selected groups; or one selected from the group consisting of a C1-C3 alkyl group, a C2-C4 alkoxycarbonyl group, and a halogen atom; Indicates a pyrrolidinylcarbonyl group that may be substituted with two or more types of groups, and when there are multiple groups, they may be the same or different,
n represents an integer from 0 to 4,
p represents 0 or 1, and q represents 0 or 1,
Pharmaceutical composition according to [3].
［５］
　前記TRPC3/6/7チャネル活性化物質が、前記式（１）で表わされる化合物であって、
　Ａは、窒素原子を示し、
　Ｂは、炭素原子または窒素原子を示し、
　Ｒ１は、フッ素原子、塩素原子、Ｃ１～Ｃ４アルキル基、Ｃ１～Ｃ４アルコキシ基、Ｃ２～Ｃ４アルコキシカルボニル基、トリフルオロメチル基、トリクロロメチル基、またはトリブロモメチル基を示し、複数あるときは同一または異なってよく、
　ｍは、０～３の整数を示し、
　Ｒ２は、フッ素原子；塩素原子；トリフルオロメチル基；トリクロロメチル基；Ｃ１～Ｃ４アルキル基；Ｃ２～Ｃ４アシルオキシ基；Ｃ２～Ｃ４アルコキシカルボニル基；Ｃ１～Ｃ３アルキル基およびＣ２～Ｃ４アルコキシカルボニル基からなる群から選択される１種または２種の基で置換されてもよいアミド基；Ｃ１～Ｃ３アルキル基、Ｃ２～Ｃ４アルコキシカルボニル基およびハロゲン原子からなる群から選択される１種または２種以上の基で置換されてもよいピペリジニルカルボニル基；或いはＣ１～Ｃ３アルキル基、Ｃ２～Ｃ４アルコキシカルボニル基およびハロゲン原子からなる群から選択される１種または２種以上の基で置換されてもよいピロリジニルカルボニル基を示し、複数あるときは同一または異なってよく、
　ｎは、０～３の整数を示し、
　ｐは、１を示し、そして
　ｑは、１を示す、
［４］に記載の医薬組成物。
［６］
　前記TRPC3/6/7チャネル活性化物質が、前記式（１）で表わされる化合物であって、
　Ａは、窒素原子を示し、
　Ｂは、炭素原子または窒素原子を示し、
　Ｒ１は、塩素原子、Ｃ１～Ｃ３アルキル基、Ｃ１～Ｃ３アルコキシ基、Ｃ２～Ｃ４アルコキシカルボニル基、トリフルオロメチル基、またはトリクロロメチル基を示し、複数あるときは同一または異なってよく、
　ｍは、０～２の整数を示し、
　Ｒ２は、フッ素原子；塩素原子；トリフルオロメチル基；トリクロロメチル基；Ｃ１～Ｃ４アルキル基；Ｃ２～Ｃ４アルコキシカルボニル基；Ｃ１～Ｃ３アルキル基およびＣ２～Ｃ４アルコキシカルボニル基からなる群から選択される１種または２種の基で置換されてもよいアミド基；或いはＣ１～Ｃ３アルキル基、Ｃ２～Ｃ４アルコキシカルボニル基およびハロゲン原子からなる群から選択される１種または２種以上の基で置換されてもよいピペリジニルカルボニル基を示し、複数あるときは同一または異なってよく、
　ｎは、０～３の整数を示し、
　ｐは、１を示し、
　ｑは、１を示す、
［５］記載の医薬組成物。 [5]
The TRPC3/6/7 channel activator is a compound represented by the formula (1),
A represents a nitrogen atom,
B represents a carbon atom or a nitrogen atom,
R 1 represents a fluorine atom, a chlorine atom, a C1-C4 alkyl group, a C1-C4 alkoxy group, a C2-C4 alkoxycarbonyl group, a trifluoromethyl group, a trichloromethyl group, or a tribromomethyl group, and when there is more than one, may be the same or different;
m represents an integer from 0 to 3,
R 2 is a fluorine atom; a chlorine atom; a trifluoromethyl group; a trichloromethyl group; a C1 to C4 alkyl group; a C2 to C4 acyloxy group; a C2 to C4 alkoxycarbonyl group; a C1 to C3 alkyl group and a C2 to C4 alkoxycarbonyl group An amide group which may be substituted with one or two groups selected from the group consisting of; one or two groups selected from the group consisting of a C1-C3 alkyl group, a C2-C4 alkoxycarbonyl group, and a halogen atom; A piperidinylcarbonyl group which may be substituted with any of the above groups; or one or more groups selected from the group consisting of a C1-C3 alkyl group, a C2-C4 alkoxycarbonyl group, and a halogen atom. Indicates a pyrrolidinyl carbonyl group, which may be the same or different when there are multiple groups,
n represents an integer from 0 to 3;
p represents 1, and q represents 1,
Pharmaceutical composition according to [4].
[6]
The TRPC3/6/7 channel activator is a compound represented by the formula (1),
A represents a nitrogen atom,
B represents a carbon atom or a nitrogen atom,
R 1 represents a chlorine atom, a C1-C3 alkyl group, a C1-C3 alkoxy group, a C2-C4 alkoxycarbonyl group, a trifluoromethyl group, or a trichloromethyl group, and when there is more than one, they may be the same or different,
m represents an integer from 0 to 2,
R 2 is selected from the group consisting of a fluorine atom; a chlorine atom; a trifluoromethyl group; a trichloromethyl group; a C1-C4 alkyl group; a C2-C4 alkoxycarbonyl group; a C1-C3 alkyl group and a C2-C4 alkoxycarbonyl group. an amide group which may be substituted with one or two groups; or substituted with one or more groups selected from the group consisting of a C1-C3 alkyl group, a C2-C4 alkoxycarbonyl group, and a halogen atom; Indicates a piperidinyl carbonyl group that may be
n represents an integer from 0 to 3;
p indicates 1,
q indicates 1,
The pharmaceutical composition according to [5].
［７］
　前記TRPC3/6/7チャネル活性化物質が、以下に示された化合物の１種または２種以上である、［１記載の医薬組成物：
Figure JPOXMLDOC01-appb-C000009
Figure JPOXMLDOC01-appb-C000010
Figure JPOXMLDOC01-appb-C000011
 ［８］
　心不全が、急性心不全、または慢性心不全における急性増悪である、［１］記載の医薬組成物。 [7]
The pharmaceutical composition according to [1], wherein the TRPC3/6/7 channel activator is one or more of the compounds shown below:
Figure JPOXMLDOC01-appb-C000009
Figure JPOXMLDOC01-appb-C000010
Figure JPOXMLDOC01-appb-C000011
 [8]
The pharmaceutical composition according to [1], wherein the heart failure is acute heart failure or acute exacerbation of chronic heart failure.
＜スクリーニング方法＞
［９］
　心不全または骨格筋不全を予防および／または処置するための物質をスクリーニングする方法であって、TRPC3/6/7チャネル活性化物質を選別する、該スクリーニング方法。
［１０］
　被験物質の存在下で、細胞中の亜鉛濃度を測定する工程、および前記亜鉛濃度が、前記被験物質の非存在下における亜鉛濃度と比較して増加していた場合に、前記被験物質はTRPC3/6/7チャネル活性化をもたらす候補物質であると判断する工程
を備える、［８］記載のスクリーニング方法。 <Screening method>
[9]
A method of screening for substances for preventing and/or treating heart failure or skeletal muscle failure, the screening method comprising selecting TRPC3/6/7 channel activators.
[10]
measuring the zinc concentration in cells in the presence of a test substance, and if the zinc concentration is increased compared to the zinc concentration in the absence of the test substance, the test substance The screening method according to [8], comprising the step of determining that the substance is a candidate substance that causes 6/7 channel activation.
＜化合物＞
［１１］
　式（１）：
Figure JPOXMLDOC01-appb-C000012
 ［式中、
　Ａは、炭素原子または窒素原子を示し、
　Ｂは、炭素原子または窒素原子を示し、
　Ｒ１は、ハロゲン原子、アルキル基、アルコキシ基、アシルオキシ基、アルコキシカルボニル基、またはハロゲン原子で置換されたアルキル基を示し、複数あるときは同一または異なってよく、
　ｍは、０～５の整数を示し、
　ｎは、２～５の整数を示し、
　複数あるＲ２のうち、少なくとも１つはハロゲン原子、アルキル基またはトリフルオロメチル基であり；残りのＲ２は、１）単数または複数のハロゲン原子で置換され、かつアルキル基およびアルコキシカルボニル基からなる群から選択される１種または２種以上の基で置換されてもよいピペリジニルカルボニル基；或いは２）単数または複数のハロゲン原子で置換され、かつアルキル基およびアルコキシカルボニル基からなる群から選択される１種または２種以上の基で置換されてもよいピロリジニルカルボニル基を示し、複数あるときは同一または異なってよく、
　ｐは、０または１を示し、そして
　ｑは、０または１を示す。］
で表される化合物。 <Compound>
[11]
Formula (1):
Figure JPOXMLDOC01-appb-C000012
 [In the formula,
A represents a carbon atom or a nitrogen atom,
B represents a carbon atom or a nitrogen atom,
R 1 represents a halogen atom, an alkyl group, an alkoxy group, an acyloxy group, an alkoxycarbonyl group, or an alkyl group substituted with a halogen atom, and when there is more than one, they may be the same or different;
m represents an integer from 0 to 5,
n represents an integer from 2 to 5,
At least one of the plurality of R 2 is a halogen atom, an alkyl group, or a trifluoromethyl group; the remaining R 2 are 1) substituted with a single or multiple halogen atoms, and are selected from an alkyl group and an alkoxycarbonyl group. or 2) a piperidinylcarbonyl group optionally substituted with one or more groups selected from the group consisting of alkyl groups and alkoxycarbonyl groups; or 2) substituted with one or more halogen atoms; Indicates a pyrrolidinyl carbonyl group that may be substituted with one or more selected groups, and when there are multiple groups, they may be the same or different,
p represents 0 or 1, and q represents 0 or 1. ]
A compound represented by
　心不全治療の際、最も重要視される「血圧低下作用」はTRPC6活性化では起こりえないため、本創薬戦略が有望視される。 This drug discovery strategy is seen as promising because the "blood pressure lowering effect", which is most important in the treatment of heart failure, cannot occur through TRPC6 activation.
図１は、野生型（WT）およびTRPC6遺伝子欠損マウス（TRPC6(-/-)）において血管拡張薬ヒドララジン（Hyd）静脈内投与による左心室収縮力の増加分および心拍数の増加分を示すグラフである。ΔLVdP/dtmax：左心室収縮力の増加分、ΔHeart Rate：心拍数の増加分、AUC：area under the curve（投与後10分間の増加分の総和面積）。**; P<0.01.Figure 1 is a graph showing the increase in left ventricular contractility and heart rate due to intravenous administration of the vasodilator hydralazine (Hyd) in wild type (WT) and TRPC6 gene-deficient mice (TRPC6(-/-)). It is. ΔLVdP/dtmax: increase in left ventricular contractile force, ΔHeart Rate: increase in heart rate, AUC: area under the curve (total area under the curve for 10 minutes after administration). **; P<0.01.
図２は、MLP欠損マウスにおけるPPZ2の心不全進行の抑制効果を示す。図２a：左心室駆出率（ejection fraction、EF）を示すグラフである。図２b：左心室内径短縮率（fractional shortening、FS）を示すグラフである。図２c：HE染色像の写真（左）、およびそれから算出される心筋細胞面積（CSA；cross sectional area）を示すグラフである（右）。スケールバー = 25 μm。　 *; P<0.05. Vehicle群：N＝5、PPZ2群：N=6。図２d：シリウスレッド染色画像（左）およびそれから算出される陽性コラーゲン発現部位（CVF；コラーゲン容量画分（collagen volume fraction））を示すグラフである。スケールバー = 100 μm。*; P<0.05. Vehicle群：N＝5、PPZ2群：N=6。Figure 2 shows the inhibitory effect of PPZ2 on the progression of heart failure in MLP-deficient mice. Figure 2a: Graph showing left ventricular ejection fraction (EF). Figure 2b: A graph showing fractional shortening (FS) of the left ventricle. Figure 2c: A photograph of the HE staining image (left) and a graph showing the cardiomyocyte area (CSA; cross sectional area) calculated from it (right). Scale bar = 25 μm. *; P<0.05. Vehicle group: N=5, PPZ2 group: N=6. FIG. 2d: Graph showing the Sirius red stained image (left) and the positive collagen expression site (CVF; collagen volume fraction) calculated from it. Scale bar = 100 μm. *; P<0.05. Vehicle group: N=5, PPZ2 group: N=6.
図３－１は、圧負荷誘発心不全マウスにおけるPPZ2の心不全改善作用を示す。図３－１a：左心室駆出率（ejection fraction、EF）を示すグラフである。図３－１b：左心室内径短縮率（fractional shortening、FS）を示すグラフである。*; P<0.05. N＝5。図３－１c：WGA染色像の写真（左）、およびそれから算出される心筋細胞面積（CSA；cross sectional area）を示すグラフである（右）。スケールバー = 25 μm。　 *; P<0.05. Vehicle群：N＝5、PPZ2群：N=6。Figure 3-1 shows the heart failure improving effect of PPZ2 in mice with pressure overload-induced heart failure. Figure 3-1a: A graph showing left ventricular ejection fraction (EF). Figure 3-1b: A graph showing fractional shortening (FS) of the left ventricle. *; P<0.05. N=5. Figure 3-1c: A photograph of a WGA-stained image (left) and a graph showing the cardiomyocyte area (CSA; cross sectional area) calculated from it (right). Scale bar = 25 μm. *; P<0.05. Vehicle group: N=5, PPZ2 group: N=6. 図３－２は、圧負荷誘発心不全マウスにおけるPPZ2の心不全改善作用を示す。図３－２d：シリウスレッド染色画像（左）およびそれから算出される陽性コラーゲン発現部位（CVF；コラーゲン容量画分（collagen volume fraction））を示すグラフである。スケールバー = 100 μm。*; P<0.05. Vehicle群：N＝5、PPZ2群：N=6。図３－２e：心筋組織中のZn2+濃度（Zinpyr-1染色）を示す写真である。図３－２f：プロテインキナーゼG（PKG）活性を示すグラフである。対照; n=5, PPZ2; n=5. *; P<0.05, **; P<0.01, ***; P<0.001, ****; P<0.0001。Figure 3-2 shows the heart failure ameliorating effect of PPZ2 in mice with pressure overload-induced heart failure. FIG. 3-2d: A graph showing the Sirius red stained image (left) and the positive collagen expression site (CVF; collagen volume fraction) calculated from it. Scale bar = 100 μm. *; P<0.05. Vehicle group: N=5, PPZ2 group: N=6. Figure 3-2e: Photograph showing Zn 2+ concentration (Zinpyr-1 staining) in myocardial tissue. Figure 3-2f: Graph showing protein kinase G (PKG) activity. Control; n=5, PPZ2; n=5. *; P<0.05, **; P<0.01, ***; P<0.001, ****; P<0.0001.
図４は、MIモデルマウスにおけるPPZ2の心不全抑制効果を示すグラフである。N=5.FIG. 4 is a graph showing the heart failure suppressive effect of PPZ2 in MI model mice. N=5.
図５は、PPZ2投与による正常マウスの圧受容反射応答（陽性変力作用）の増強の用量依存性とTRPC6阻害剤BI749327による抑制効果を示すグラフである。縦軸はヒドララジン誘発性の（圧受容反射による）左心室収縮力の増加分（ΔLVdP/dtmax）または心拍数の増加分（ΔHeart Rate）を示している。Δ値はヒドララジン（Hyd）投与前の値と投与10分間での最大値との差分を示す。FIG. 5 is a graph showing the dose dependence of enhancement of the baroreceptor reflex response (positive inotropic effect) in normal mice by administration of PPZ2 and the suppressive effect of the TRPC6 inhibitor BI749327. The vertical axis shows the hydralazine-induced increase in left ventricular contractile force (ΔLVdP/dtmax) or heart rate (ΔHeart Rate). The Δ value indicates the difference between the value before hydralazine (Hyd) administration and the maximum value 10 minutes after administration.
図６は、TRPC6発現HEK293細胞を用いたTRPC6活性化物質による細胞内Zn2+濃度変化を示す。図６a、図６b：PPZ2 (30μM)処置によるFluoZn-3蛍光強度変化とその増加率（AUC; area under the curve）を比較しているグラフである。図６c、図６d：TRPC6チャネル活性化物質GSK1702934A (10μM)処置によるFluoZn-3蛍光強度変化とその増加率を比較しているグラフである。N=3。FIG. 6 shows changes in intracellular Zn 2+ concentration caused by TRPC6 activators using TRPC6-expressing HEK293 cells. Figures 6a and 6b are graphs comparing the FluoZn-3 fluorescence intensity change and its increase rate (AUC; area under the curve) due to PPZ2 (30 μM) treatment. FIG. 6c, FIG. 6d: Graphs comparing FluoZn-3 fluorescence intensity change and its increase rate due to treatment with TRPC6 channel activator GSK1702934A (10 μM). N=3.
図７は、新生仔ラット心筋細胞を用いたTRPC6活性化物質による心筋収縮力増強作用を検討した結果である。図７a：ノルエピネフリン（NE）（0.1μM）または亜鉛のキレーターであるTPEN (N,N,N',N'-テトラキス(2-ピリジルメチルエチレンジアミンによる心筋細胞収縮に対するPPZ2 (30μM)処置の効果を示すグラフである。図７b：NE（1μM）による心筋細胞収縮に対するGSK1702934A(1μM)またはハイパーフォリン（hyperforin）(1μM)処置の効果を示すグラフである。FIG. 7 shows the results of examining the myocardial contractility enhancing effect of TRPC6 activators using neonatal rat cardiomyocytes. Figure 7a: Showing the effect of PPZ2 (30 μM) treatment on cardiomyocyte contraction induced by norepinephrine (NE) (0.1 μM) or the zinc chelator TPEN (N,N,N',N'-tetrakis(2-pyridylmethylethylenediamine). Figure 7b: Graph showing the effect of GSK1702934A (1 μM) or hyperforin (1 μM) treatment on cardiomyocyte contraction induced by NE (1 μM). 図８はPPZ2によって筋ジストロフィーモデルマウスにおける筋障害マーカー（左：LDH、右：CK）の増加が抑制されたことを示すグラフである。FIG. 8 is a graph showing that PPZ2 suppressed the increase in myopathy markers (left: LDH, right: CK) in muscular dystrophy model mice. 図９は、ヒトTRPC6チャネルを発現したHEK293細胞内へのCa２＋の流入に対するL687およびPPZ2の作用を示すグラフである。FIG. 9 is a graph showing the effects of L687 and PPZ2 on Ca 2+ influx into HEK293 cells expressing human TRPC6 channels. 図１０は、ヒトTRPC6チャネルを発現したHEK293細胞内へのCa２＋の流入に対する本発明化合物の作用を示すグラフである。FIG. 10 is a graph showing the effect of the compounds of the present invention on Ca 2+ influx into HEK293 cells expressing human TRPC6 channels. 図１１は、ヒトTRPC3チャネルを発現したHEK293細胞内へのCa２＋の流入に対する本発明化合物の作用を示すグラフである。FIG. 11 is a graph showing the effect of the compounds of the present invention on Ca 2+ influx into HEK293 cells expressing human TRPC3 channels. 図１２上段は、ラット新生児心室筋細胞のαアクチニン染色像、下段は、エンドセリン－１（ET-1） 100 nMを４８時間刺激させた後の心筋細胞面積の増大（心肥大）に対するL-0044 (10 μM)の肥大抑制作用を示す。下段は心筋細胞面積の定量結果を示すグラフである。Figure 12. Upper panel shows α-actinin staining images of neonatal rat ventricular myocytes. Lower panel shows L-0044 response to increase in cardiomyocyte area (cardiac hypertrophy) after stimulation with 100 nM endothelin-1 (ET-1) for 48 hours. (10 μM) shows hypertrophic inhibitory effect. The lower row is a graph showing the quantitative results of myocardial cell area. 図１３は、mCherry蛍光タンパク質を付加したマウスTRPC6チャネルを発現したHEK293細胞（右）と何も発現させていないHEK293細胞（左）へのZn2+の流入に対する本発明化合物の作用を示すグラフである。縦軸はFluoZn-3蛍光強度変化量を示す。Figure 13 is a graph showing the effect of the compound of the present invention on Zn 2+ influx into HEK293 cells expressing the mouse TRPC6 channel with mCherry fluorescent protein added (right) and HEK293 cells not expressing anything (left). be. The vertical axis indicates the amount of FluoZn-3 fluorescence intensity change. 図１４は、ノルエピネフリン（NE）（0.1μM）刺激による心筋細胞の収縮に対するL-0044 (1μM)処置の効果を示すグラフである。FIG. 14 is a graph showing the effect of L-0044 (1 μM) treatment on the contraction of cardiomyocytes stimulated by norepinephrine (NE) (0.1 μM).
＜医薬組成物＞
　本発明はひとつの形態において、受容体作動性TRPC3/6/7チャネルの活性化を促す低分子化合物が、降圧作用を持たずに急性心不全や慢性心不全の急性増悪に有効な治療薬となることを示す用途発明である。心不全の状態は、心臓から血液を送り出す力が元来、弱くなっているため、血圧が上がりにくい状態にある。ここに降圧作用が加わると慢性低血圧状態となり、血圧を上げようとする代償機構、すなわち交感神経活動の亢進が働く。これが結果的に、慢性心不全を悪化させる原因になるため、降圧作用を有さない点は、本発明の大きな利点である。
　本発明は、その実施態様において、TRPC3/6/7チャネル活性化物質、具体的にはピペラジン誘導体（PPZ）を含有する、心不全または骨格筋不全を予防および／または処置するための医薬組成物を提供する。 <Pharmaceutical composition>
In one form, the present invention provides that a low-molecular compound that promotes activation of receptor-operated TRPC3/6/7 channels becomes an effective therapeutic agent for acute heart failure or acute exacerbation of chronic heart failure without having a hypotensive effect. This is a use invention showing the following. In a state of heart failure, the ability of the heart to pump blood is originally weakened, making it difficult for blood pressure to rise. When antihypertensive effects are added here, a state of chronic hypotension occurs, and a compensatory mechanism that attempts to raise blood pressure, ie, an increase in sympathetic nerve activity, comes into play. This ultimately causes worsening of chronic heart failure, so the fact that it does not have a hypotensive effect is a major advantage of the present invention.
In its embodiments, the present invention provides a pharmaceutical composition for preventing and/or treating heart failure or skeletal muscle failure, containing a TRPC3/6/7 channel activator, specifically a piperazine derivative (PPZ). provide.
　本発明者らは、TRPC6遺伝子欠損マウスが心不全に対して脆弱な傾向を示すこと、すなわちTRPC6欠損マウスの心臓が高血圧負荷に対して酸化ストレスや炎症増加を起こし易く、高血圧負荷に適応できていないことに注目した。交感神経終末から放出されるノルアドレナリンは、心臓のポンプ機能を増強させる主要な神経伝達物質であり、特に急な血圧低下に対する代償的な心機能亢進（圧受容反射）に重要な役割を果たしている。ノルアドレナリンは、心筋細胞膜上のβアドレナリン受容体を活性化することで、心筋細胞の収縮や拍動数の上昇を引き起こす。心不全時には血中ノルアドレナリン濃度が上昇するにも関わらず、ポンプ機能が十分に働かなくなっており、この原因として、βアドレナリン受容体数の脱感作や受容体数低下が指摘されている。TRPC6欠損マウスでは、野生型マウスと比べて、血圧降下薬ヒドララジン誘発性の圧受容反射応答（陽性変時作用（心拍数の増加）と陽性変力作用（収縮力の増加））のうち、陽性変力作用が著しく減弱していた（図１）。心不全においても圧受容反射応答が著しく減弱しており、予後悪化の原因となることから、TRPC6チャネル活性化が心不全の改善に働く可能性が示された。本発明者らは、この関係に着目し、TRPC3/6/7チャネルの活性化と心不全または骨格筋不全の予防または処置との関係を初めて見出した。 The present inventors demonstrated that TRPC6 gene-deficient mice show a tendency to be vulnerable to heart failure, that is, the hearts of TRPC6-deficient mice are susceptible to increased oxidative stress and inflammation in response to hypertensive stress, and are unable to adapt to hypertensive stress. I paid attention to that. Norepinephrine released from sympathetic nerve terminals is a major neurotransmitter that enhances the pumping function of the heart, and plays an especially important role in compensatory cardiac hyperactivity (baroreceptive reflex) in response to sudden drops in blood pressure. Noradrenaline activates β-adrenergic receptors on the cardiac muscle cell membrane, causing contraction of the cardiac muscle cell and an increase in heart rate. Despite the increase in blood noradrenaline concentration during heart failure, the pump function does not function sufficiently, and desensitization of the number of β-adrenergic receptors and a decrease in the number of receptors have been pointed out as the cause of this. Compared to wild-type mice, TRPC6-deficient mice showed positive baroreflex responses (positive chronotropy (increased heart rate) and positive inotropy (increased contractile force)) induced by the antihypertensive drug hydralazine. The inotropic effect was significantly attenuated (Figure 1). The baroreceptive reflex response is also markedly attenuated in heart failure, which is a cause of worsening prognosis, suggesting that TRPC6 channel activation may work to improve heart failure. The present inventors focused on this relationship and found for the first time a relationship between activation of TRPC3/6/7 channels and prevention or treatment of heart failure or skeletal muscle failure.
　以下に記載する試験例１－３では、異なる実験動物を利用し、モデルの違いや種差・系統差によって表現型が変わることが少なからず生じるにもかかわらず、異なる心不全モデル、異なるマウス系統種であっても同様に心不全保護効果があること（共通性・普遍性）を示すことができた。 In Test Examples 1-3 described below, different experimental animals were used, and despite the fact that the phenotype often changes due to model differences, species differences, and strain differences, different heart failure models and different mouse strains were used. We were able to show that the same protective effect against heart failure exists (commonality/universality).
　本発明において、「TRPC3/6/7チャネル活性化物質」とは、ホスホリパーゼC連関型受容体刺激によって活性化されるTRPCチャネルのうち、ジアシルグリセロールで直接的に活性化されるチャネル群（TRPC3/6/7）を、ジアシルグリセロールと関係なく開口活性化させる物質をいう。そして、「TRPC3/6/7チャネル活性化物質」は、TRPC3、TRPC6およびTRPC7のうちどれか、またはそれらすべてを活性化する物質群を意味する。 In the present invention, "TRPC3/6/7 channel activator" refers to a group of channels that are directly activated by diacylglycerol (TRPC3/ 6/7), which activates the opening of diacylglycerol independently of diacylglycerol. "TRPC3/6/7 channel activator" means a group of substances that activate any or all of TRPC3, TRPC6, and TRPC7.
　本発明において、「TRPC3/6/7チャネル活性化物質」には、ピペラジン誘導体（PPZ）が含まれ、その具体例として、Sawamura S, Hatano M, Takada Y, Hino K, Kawamura T, Tanikawa J, Nakagawa H, Hase H, Nakao A, Hirano M, Rotrattanadumrong R, Kiyonaka S, Mori MX, Nishida M, Hu Y, Inoue R, Nagata R, Mori Y. Screening of Transient Receptor Potential Canonical Channel Activators Identifies Novel Neurotrophic Piperazine Compounds. Mol Pharmacol. 89(3), 348-363 (2016). doi: 10.1124/mol.115.102863に記載されているピペラジン誘導体が挙げられる。例えば、式：
Figure JPOXMLDOC01-appb-C000013
 で示されるいずれかの化合物が挙げられる。 In the present invention, the "TRPC3/6/7 channel activator" includes piperazine derivatives (PPZ), and specific examples thereof include Sawamura S, Hatano M, Takada Y, Hino K, Kawamura T, Tanikawa J, Nakagawa H, Hase H, Nakao A, Hirano M, Rotrattanadumrong R, Kiyonaka S, Mori MX, Nishida M, Hu Y, Inoue R, Nagata R, Mori Y. Screening of Transient Receptor Potential Canonical Channel Activators Identifies Novel Neurotrophic Piperazine Compounds. Examples include piperazine derivatives described in Mol Pharmacol. 89(3), 348-363 (2016). doi: 10.1124/mol.115.102863. For example, the formula:
Figure JPOXMLDOC01-appb-C000013
 Examples include any of the compounds shown below.
　また、本発明において、「TRPC3/6/7チャネル活性化物質」には、Xu X, Lozinskaya I, Costell M, Lin Z, Ball JA, Bernard R, Behm DJ, Marino JP, Schnackenberg (2013). "CG Characterization of Small Molecule TRPC3 and TRPC6 agonist and Antagonists". Biophysical Journal. 104: 454a. doi:10.1016/j.bpj.2012.11.2513.に記載されている、以下の式：
Figure JPOXMLDOC01-appb-C000014
 で示されるGSK1702934Aが包含される。 Furthermore, in the present invention, "TRPC3/6/7 channel activator" includes Xu X, Lozinskaya I, Costell M, Lin Z, Ball JA, Bernard R, Behm DJ, Marino JP, Schnackenberg (2013). The following formula described in "CG Characterization of Small Molecule TRPC3 and TRPC6 agonist and Antagonists". Biophysical Journal. 104: 454a. doi:10.1016/j.bpj.2012.11.2513.
Figure JPOXMLDOC01-appb-C000014
 Includes GSK1702934A shown in
　さらに、本発明において、「TRPC3/6/7チャネル活性化物質」には、TRPV1チャネル、TRPV2チャネルおよびTRPA1チャネルを活性化することが知られているカンナビジオール（Cannabidiol、N. Qin et al., J. Neurosci. 28, 6231-6238 (2008)）も包含される。 Furthermore, in the present invention, "TRPC3/6/7 channel activator" includes cannabidiol, N. Qin et al., which is known to activate TRPV1 channel, TRPV2 channel, and TRPA1 channel. J. Neurosci. 28, 6231-6238 (2008)) is also included.
　さらにまた、本発明において、「TRPC3/6/7チャネル活性化物質」には、WO2022/118966に記載されている化合物を包含する化合物が含まれる。 Furthermore, in the present invention, "TRPC3/6/7 channel activator" includes compounds including the compounds described in WO2022/118966.
　すなわち、本発明は第一の形態として、TRPC3/6/7チャネル活性化物質を含有する、心不全または骨格筋不全を予防および／または処置するための医薬組成物；
　好ましくは、TRPC3/6/7チャネル活性化物質が、式（１）：
Figure JPOXMLDOC01-appb-C000015
 ［式中、
　Ａは、炭素原子または窒素原子を示し、
　Ｂは、炭素原子または窒素原子を示し、
　Ｒ１は、置換されてもよいアルキル基を示し、複数あるときは同一または異なってよく、
　ｍは、０～５の整数を示し、
　Ｒ２は、ハロゲン原子；トリハロゲノメチル基；アルキル基；アシルオキシ基；アルコキシカルボニル基；置換されてもよいアミド基；置換されてもよいピペリジニルカルボニル基；或いは置換されてもよいピロリジニルカルボニル基を示し、複数あるときは同一または異なってよく、
　ｎは、０～５の整数を示し、
　ｐは、０または１を示し、そして
　ｑは、０または１を示す。］
で表される化合物、ハイパーフォリン、GSK1702934A、およびカンナビジオール；
からなる群から選択される少なくとも１種の化合物である、本発明の医薬組成物； That is, the present invention, as a first aspect, provides a pharmaceutical composition for preventing and/or treating heart failure or skeletal muscle failure, which contains a TRPC3/6/7 channel activator;
Preferably, the TRPC3/6/7 channel activator has the formula (1):
Figure JPOXMLDOC01-appb-C000015
 [In the formula,
A represents a carbon atom or a nitrogen atom,
B represents a carbon atom or a nitrogen atom,
R 1 represents an optionally substituted alkyl group, and when there is a plurality of them, they may be the same or different,
m represents an integer from 0 to 5,
R 2 is a halogen atom; a trihalogenomethyl group; an alkyl group; an acyloxy group; an alkoxycarbonyl group; an optionally substituted amide group; an optionally substituted piperidinylcarbonyl group; or an optionally substituted pyrrolidinyl Indicates a carbonyl group, and when there are multiple groups, they may be the same or different,
n represents an integer from 0 to 5,
p represents 0 or 1, and q represents 0 or 1. ]
Compounds represented by, hyperforin, GSK1702934A, and cannabidiol;
The pharmaceutical composition of the present invention is at least one compound selected from the group consisting of;
　より好ましくは、TRPC3/6/7チャネル活性化物質が、式（１）：
Figure JPOXMLDOC01-appb-C000016
 ［式中、
　Ａは、炭素原子または窒素原子を示し、
　Ｂは、炭素原子または窒素原子を示し、
　Ｒ１は、ハロゲン原子、アルキル基、アルコキシ基、アシルオキシ基、アルコキシカルボニル基、またはハロゲン原子で置換されたアルキル基を示し、複数あるときは同一または異なってよく、
　ｍは、０～５の整数を示し、
　Ｒ２は、ハロゲン原子；トリハロゲノメチル基；アルキル基；アシルオキシ基；アルコキシカルボニル基；アルキル基、アルコキシカルボニル基およびハロゲン原子、好ましくはフッ素原子からなる群から選択される１種または２種の基で置換されてもよいアミド基；アルキル基、アルコキシカルボニル基およびハロゲン原子、好ましくはフッ素原子からなる群から選択される１種または２種以上の基で置換されてもよいピペリジニルカルボニル基；或いはアルキル基、アルコキシカルボニル基およびハロゲン原子、好ましくはフッ素原子からなる群から選択される１種または２種以上の基で置換されてもよいピロリジニルカルボニル基を示し、複数あるときは同一または異なってよく、
　ｎは、０～５の整数を示し、
　ｐは、０または１を示し、
ｑは、０または１を示す。］
で表される化合物、ハイパーフォリン、GSK1702934A、およびカンナビジオール；
からなる群から選択される少なくとも１種の化合物である、本発明の医薬組成物に関する。 More preferably, the TRPC3/6/7 channel activator has the formula (1):
Figure JPOXMLDOC01-appb-C000016
 [In the formula,
A represents a carbon atom or a nitrogen atom,
B represents a carbon atom or a nitrogen atom,
R 1 represents a halogen atom, an alkyl group, an alkoxy group, an acyloxy group, an alkoxycarbonyl group, or an alkyl group substituted with a halogen atom, and when there is more than one, they may be the same or different;
m represents an integer from 0 to 5,
R 2 is one or two groups selected from the group consisting of a halogen atom; a trihalogenomethyl group; an alkyl group; an acyloxy group; an alkoxycarbonyl group; an alkyl group, an alkoxycarbonyl group, and a halogen atom, preferably a fluorine atom. An amide group which may be substituted with; a piperidinylcarbonyl group which may be substituted with one or more groups selected from the group consisting of an alkyl group, an alkoxycarbonyl group, and a halogen atom, preferably a fluorine atom; Alternatively, it represents a pyrrolidinylcarbonyl group which may be substituted with one or more groups selected from the group consisting of an alkyl group, an alkoxycarbonyl group, and a halogen atom, preferably a fluorine atom, and when there are multiple groups, they are the same or It's okay to be different,
n represents an integer from 0 to 5,
p represents 0 or 1;
q represents 0 or 1. ]
Compounds represented by, hyperforin, GSK1702934A, and cannabidiol;
The pharmaceutical composition of the present invention is at least one compound selected from the group consisting of:
　さらにより好ましくは、本発明は、TRPC3/6/7チャネル活性化物質が、前記式（１）で表わされる化合物であって、
　Ａは、炭素原子または窒素原子を示し、
　Ｂは、炭素原子または窒素原子を示し、
　Ｒ１は、フッ素原子、塩素原子、臭素原子、Ｃ１～Ｃ４アルキル基、Ｃ１～Ｃ４アルコキシ基、Ｃ２～Ｃ５アシルオキシ基、Ｃ２～Ｃ５アルコキシカルボニル基、またはトリハロゲノメチル基を示し、複数あるときは同一または異なってよく、
　ｍは、０～４の整数を示し、
　Ｒ２は、フッ素原子；塩素原子；臭素原子；トリフルオロメチル基；トリクロロメチル基；トリブロモメチル基；Ｃ１～Ｃ４アルキル基；Ｃ２～Ｃ５アシルオキシ基；Ｃ２～Ｃ５アルコキシカルボニル基；Ｃ１～Ｃ３アルキル基およびＣ２～Ｃ４アルコキシカルボニル基からなる群から選択される１種または２種の基で置換されてもよいアミド基；Ｃ１～Ｃ３アルキル基、Ｃ２～Ｃ４アルコキシカルボニル基およびハロゲン原子、好ましくはフッ素原子からなる群から選択される１種または２種以上の基で置換されてもよいピペリジニルカルボニル基；或いはＣ１～Ｃ３アルキル基、Ｃ２～Ｃ４アルコキシカルボニル基、およびハロゲン原子からなる群から選択される１種または２種以上の基で置換されてもよいピロリジニルカルボニル基を示し、複数あるときは同一または異なってよく、
　ｎは、０～４の整数を示し、
　ｐは、０または１を示し、そして
　ｑは、０または１を示す、本発明の医薬組成物； Even more preferably, the present invention provides that the TRPC3/6/7 channel activator is a compound represented by the above formula (1),
A represents a carbon atom or a nitrogen atom,
B represents a carbon atom or a nitrogen atom,
R 1 represents a fluorine atom, a chlorine atom, a bromine atom, a C1-C4 alkyl group, a C1-C4 alkoxy group, a C2-C5 acyloxy group, a C2-C5 alkoxycarbonyl group, or a trihalogenomethyl group, and when there is more than one, may be the same or different;
m represents an integer from 0 to 4,
R 2 is a fluorine atom; a chlorine atom; a bromine atom; a trifluoromethyl group; a trichloromethyl group; a tribromomethyl group; a C1 to C4 alkyl group; a C2 to C5 acyloxy group; a C2 to C5 alkoxycarbonyl group; a C1 to C3 alkyl and an amide group which may be substituted with one or two groups selected from the group consisting of a C2-C4 alkoxycarbonyl group; a C1-C3 alkyl group, a C2-C4 alkoxycarbonyl group and a halogen atom, preferably fluorine a piperidinylcarbonyl group optionally substituted with one or more groups selected from the group consisting of atoms; or selected from the group consisting of a C1-C3 alkyl group, a C2-C4 alkoxycarbonyl group, and a halogen atom represents a pyrrolidinylcarbonyl group that may be substituted with one or more groups, and when there are multiple groups, they may be the same or different,
n represents an integer from 0 to 4,
The pharmaceutical composition of the present invention, wherein p represents 0 or 1, and q represents 0 or 1;
　さらに好ましくは、前記TRPC3/6/7チャネル活性化物質が、前記式（１）で表わされる化合物であって、
　Ａは、窒素原子を示し、
　Ｂは、炭素原子または窒素原子を示し、
　Ｒ１は、フッ素原子、塩素原子、Ｃ１～Ｃ４アルキル基、Ｃ１～Ｃ４アルコキシ基、Ｃ２～Ｃ４アルコキシカルボニル基、トリフルオロメチル基、トリクロロメチル基、またはトリブロモメチル基を示し、複数あるときは同一または異なってよく、
　ｍは、０～３の整数を示し、
　Ｒ２は、フッ素原子；塩素原子；トリフルオロメチル基；トリクロロメチル基；Ｃ１～Ｃ４アルキル基；Ｃ２～Ｃ４アシルオキシ基；Ｃ２～Ｃ４アルコキシカルボニル基；Ｃ１～Ｃ３アルキル基およびＣ２～Ｃ４アルコキシカルボニル基からなる群から選択される１種または２種の基で置換されてもよいアミド基；Ｃ１～Ｃ３アルキル基、Ｃ２～Ｃ４アルコキシカルボニル基およびハロゲン原子、好ましくはフッ素原子からなる群から選択される１種または２種以上の基で置換されてもよいピペリジニルカルボニル基；或いはＣ１～Ｃ３アルキル基、Ｃ２～Ｃ４アルコキシカルボニル基およびハロゲン原子、好ましくはフッ素原子からなる群から選択される１種または２種以上の基で置換されてもよいピロリジニルカルボニル基を示し、複数あるときは同一または異なってよく、
　ｎは、０～３の整数を示し、
　ｐは、１を示し、そして
　ｑは、１を示す、本発明の医薬組成物；そして More preferably, the TRPC3/6/7 channel activator is a compound represented by the formula (1),
A represents a nitrogen atom,
B represents a carbon atom or a nitrogen atom,
R 1 represents a fluorine atom, a chlorine atom, a C1-C4 alkyl group, a C1-C4 alkoxy group, a C2-C4 alkoxycarbonyl group, a trifluoromethyl group, a trichloromethyl group, or a tribromomethyl group, and when there is more than one, may be the same or different;
m represents an integer from 0 to 3,
R 2 is a fluorine atom; a chlorine atom; a trifluoromethyl group; a trichloromethyl group; a C1 to C4 alkyl group; a C2 to C4 acyloxy group; a C2 to C4 alkoxycarbonyl group; a C1 to C3 alkyl group and a C2 to C4 alkoxycarbonyl group an amide group optionally substituted with one or two groups selected from the group consisting of; a C1-C3 alkyl group, a C2-C4 alkoxycarbonyl group, and a halogen atom, preferably a fluorine atom; a piperidinylcarbonyl group which may be substituted with one or more groups; or one selected from the group consisting of a C1-C3 alkyl group, a C2-C4 alkoxycarbonyl group, and a halogen atom, preferably a fluorine atom or a pyrrolidinylcarbonyl group which may be substituted with two or more types of groups, and when there are multiple groups, they may be the same or different,
n represents an integer from 0 to 3;
The pharmaceutical composition of the present invention, wherein p represents 1 and q represents 1; and
　より好ましくは、前記TRPC3/6/7チャネル活性化物質が、前記式（１）で表わされる化合物であって、
　Ａは、窒素原子を示し、
　Ｂは、炭素原子または窒素原子を示し、
　Ｒ１は、塩素原子、Ｃ１～Ｃ３アルキル基、Ｃ１～Ｃ３アルコキシ基、Ｃ２～Ｃ４アルコキシカルボニル基、トリフルオロメチル基、またはトリクロロメチル基を示し、複数あるときは同一または異なってよく、
　ｍは、０～２の整数を示し、
　Ｒ２は、フッ素原子；塩素原子；トリフルオロメチル基；トリクロロメチル基；Ｃ１～Ｃ４アルキル基；Ｃ２～Ｃ４アルコキシカルボニル基；Ｃ１～Ｃ３アルキル基およびＣ２～Ｃ４アルコキシカルボニル基からなる群から選択される１種または２種の基で置換されてもよいアミド基；或いはＣ１～Ｃ３アルキル基、Ｃ２～Ｃ４アルコキシカルボニル基およびハロゲン原子、好ましくはフッ素原子からなる群から選択される１種または２種以上の基で置換されてもよいピペリジニルカルボニル基を示し、複数あるときは同一または異なってよく、
　ｎは、０～３の整数を示し、
　ｐは、１を示し、
　ｑは、１を示す、本発明の医薬組成物に関する。 More preferably, the TRPC3/6/7 channel activator is a compound represented by the formula (1),
A represents a nitrogen atom,
B represents a carbon atom or a nitrogen atom,
R 1 represents a chlorine atom, a C1-C3 alkyl group, a C1-C3 alkoxy group, a C2-C4 alkoxycarbonyl group, a trifluoromethyl group, or a trichloromethyl group, and when there is more than one, they may be the same or different,
m represents an integer from 0 to 2,
R 2 is selected from the group consisting of a fluorine atom; a chlorine atom; a trifluoromethyl group; a trichloromethyl group; a C1-C4 alkyl group; a C2-C4 alkoxycarbonyl group; a C1-C3 alkyl group and a C2-C4 alkoxycarbonyl group. an amide group which may be substituted with one or two groups; or one or two selected from the group consisting of a C1-C3 alkyl group, a C2-C4 alkoxycarbonyl group, and a halogen atom, preferably a fluorine atom; Indicates a piperidinylcarbonyl group that may be substituted with the above groups, and when there are multiple groups, they may be the same or different,
n represents an integer from 0 to 3;
p indicates 1,
q refers to the pharmaceutical composition of the present invention, wherein q represents 1.
　式（１）において、Ａは、炭素原子または窒素原子を示し、Ｂは炭素原子または窒素原子を示す。ＡおよびＢの組み合わせとしては、ＡおよびＢがともに炭素原子、Ａが炭素原子およびＢが窒素原子、Ａが窒素原子およびＢが炭素原子、ＡおよびＢがともに窒素原子のいずれかである。つまりＡおよびＢを含有する環は、式（１）において、ベンゼン環、ピリジン環、またはピリダジン環、ピリジン環またはピリダジン環である。 In formula (1), A represents a carbon atom or a nitrogen atom, and B represents a carbon atom or a nitrogen atom. As a combination of A and B, A and B are both carbon atoms, A is a carbon atom and B is a nitrogen atom, A is a nitrogen atom and B is a carbon atom, and A and B are both nitrogen atoms. That is, the ring containing A and B in formula (1) is a benzene ring, a pyridine ring, a pyridazine ring, a pyridine ring, or a pyridazine ring.
　式（１）において、ＡおよびＢの組み合わせとしては、ＡおよびＢがともに炭素原子、Ａが炭素原子およびＢが窒素原子、またはＡが窒素原子およびＢが炭素原子であることが好ましく、ＡおよびＢがともに炭素原子またはＡが窒素原子およびＢが炭素原子であることがより好ましい。 In formula (1), as a combination of A and B, it is preferable that both A and B are carbon atoms, A is a carbon atom and B is a nitrogen atom, or A is a nitrogen atom and B is a carbon atom; It is more preferable that both B are carbon atoms, or A is a nitrogen atom and B is a carbon atom.
　ＡおよびＢのいずれかまたは両方が炭素原子であるときは、当該炭素原子は置換基Ｒ１を有してもよい。 When either or both of A and B is a carbon atom, the carbon atom may have a substituent R 1 .
　Ｒ１が複数あるときは同一であっても異なってもよい。Ｒ１としては、フッ素原子、塩素原子、臭素原子、Ｃ１～Ｃ４アルキル基、Ｃ１～Ｃ４アルコキシ基、Ｃ２～Ｃ５アシルオキシ基、Ｃ２～Ｃ５アルコキシカルボニル基、またはトリハロゲノメチル基が好ましく、フッ素原子、塩素原子、Ｃ１～Ｃ４アルキル基、Ｃ１～Ｃ４アルコキシ基、Ｃ２～Ｃ４アルコキシカルボニル基、トリフルオロメチル基、トリクロロメチル基、またはトリブロモメチル基がより好ましく、塩素原子、Ｃ１～Ｃ３アルキル基、Ｃ１～Ｃ３アルコキシ基、Ｃ２～Ｃ４アルコキシカルボニル基、トリフルオロメチル基、またはトリクロロメチル基がさらに好ましく、塩素原子、メトキシ基、エトキシ基、メトキシカルボニル基、またはエトキシカルボニル基が特に好ましい。 When there is a plurality of R 1 's, they may be the same or different. R 1 is preferably a fluorine atom, a chlorine atom, a bromine atom, a C1-C4 alkyl group, a C1-C4 alkoxy group, a C2-C5 acyloxy group, a C2-C5 alkoxycarbonyl group, or a trihalogenomethyl group, and a fluorine atom, A chlorine atom, a C1-C4 alkyl group, a C1-C4 alkoxy group, a C2-C4 alkoxycarbonyl group, a trifluoromethyl group, a trichloromethyl group, or a tribromomethyl group are more preferable, and a chlorine atom, a C1-C3 alkyl group, a C1 -C3 alkoxy group, C2-C4 alkoxycarbonyl group, trifluoromethyl group, or trichloromethyl group is more preferred, and chlorine atom, methoxy group, ethoxy group, methoxycarbonyl group, or ethoxycarbonyl group is particularly preferred.
　Ｒ１は、Ｒ１が結合する環がベンゼン環であるときは、ベンゼン環を構成する炭素原子のうち、（ＮＨ）ｐを構成する窒素原子と直接結合する炭素原子に対してオルト位に位置する２個の炭素原子のうちの少なくとも１個に結合していることが好ましい。オルト位に位置する炭素原子に結合するＲ１としては、例えばアルコキシ基またはアルコキシカルボニル基が挙げられ、Ｃ２～Ｃ５アルコキシカルボニル基が好ましく、Ｃ２～Ｃ４アルコキシカルボニル基がより好ましく、メトキシカルボニル基またはエトキシカルボニル基がさらに好ましく、エトキシカルボニル基が特に好ましい。 When the ring to which R 1 is bonded is a benzene ring, R 1 is located at the ortho position to the carbon atom directly bonding to the nitrogen atom constituting (NH) p among the carbon atoms constituting the benzene ring. Preferably, it is bonded to at least one of two carbon atoms. Examples of R 1 bonded to the carbon atom located at the ortho position include an alkoxy group or an alkoxycarbonyl group, preferably a C2-C5 alkoxycarbonyl group, more preferably a C2-C4 alkoxycarbonyl group, and a methoxycarbonyl group or an ethoxycarbonyl group. A carbonyl group is more preferred, and an ethoxycarbonyl group is particularly preferred.
　Ｒ１は、Ｒ１が結合する環がベンゼン環であって、ベンゼン環を構成する炭素原子のうち、（ＮＨ）ｐを構成する窒素原子と直接結合する炭素原子に対してメタ位に位置する２個の炭素原子のいずれかまたは両方に結合している場合、ハロゲン原子であることが好ましく、塩素原子またはフッ素原子であることがより好ましく、フッ素原子が特に好ましい。 In R 1 , the ring to which R 1 is bonded is a benzene ring, and among the carbon atoms constituting the benzene ring, it is located at the meta position with respect to the carbon atom directly bonded to the nitrogen atom constituting (NH) p . When bonded to either or both of two carbon atoms, a halogen atom is preferable, a chlorine atom or a fluorine atom is more preferable, and a fluorine atom is particularly preferable.
　Ｒ１は、ＡおよびＢを含む環がピリジン環またはピリダジン環であるときは、ピリジン環またはピリダジン環を構成する炭素原子のうち、（ＮＨ）ｐを構成する窒素原子と直接結合する炭素原子とＡとの間に位置する炭素原子に結合することが好ましい。Ｒ１は、ＡおよびＢを含む環がピリジン環またはピリダジン環であるときは、例えばハロゲン原子、アルキル基、アルコキシ基、またはアルコキシカルボニル基であってよく、フッ素原子、塩素原子、Ｃ１～Ｃ４アルキル基、Ｃ１～Ｃ４アルコキシ基、またはＣ２～Ｃ５アルコキシカルボニル基が好ましく、Ｃ１～Ｃ４アルコキシ基がより好ましく、メトキシ基またはエトキシ基が特に好ましい。 When the ring containing A and B is a pyridine ring or a pyridazine ring, R 1 is the carbon atom that directly bonds to the nitrogen atom that constitutes (NH) p among the carbon atoms that constitute the pyridine ring or pyridazine ring. It is preferable to bond to a carbon atom located between A and A. When the ring containing A and B is a pyridine ring or a pyridazine ring, R 1 may be, for example, a halogen atom, an alkyl group, an alkoxy group, or an alkoxycarbonyl group, and may be a fluorine atom, a chlorine atom, a C1-C4 alkyl A C1-C4 alkoxy group or a C2-C5 alkoxycarbonyl group is preferred, a C1-C4 alkoxy group is more preferred, and a methoxy group or an ethoxy group is particularly preferred.
　ｍとしては、０～４の整数が好ましく、０～３の整数がより好ましく、０、１、または２がさらに好ましく、０または１がさらに好ましく、１が特に好ましい。 m is preferably an integer of 0 to 4, more preferably an integer of 0 to 3, even more preferably 0, 1, or 2, even more preferably 0 or 1, and particularly preferably 1.
　Ｒ２が複数あるときは同一であっても異なってもよい。Ｒ２としては、フッ素原子；塩素原子；臭素原子；トリフルオロメチル基；トリクロロメチル基；トリブロモメチル基；Ｃ１～Ｃ４アルキル基；Ｃ２～Ｃ５アシルオキシ基；Ｃ２～Ｃ５アルコキシカルボニル基；Ｃ１～Ｃ３アルキル基およびＣ２～Ｃ４アルコキシカルボニル基からなる群から選択される１種または２種の基で置換されてもよいアミド基；Ｃ１～Ｃ３アルキル基およびＣ２～Ｃ４アルコキシカルボニル基からなる群から選択される１種または２種以上の基で置換されてもよいピペリジニルカルボニル基；或いはＣ１～Ｃ３アルキル基およびＣ２～Ｃ４アルコキシカルボニル基からなる群から選択される１種または２種以上の基で置換されてもよいピロリジニルカルボニル基が好ましく、フッ素原子；塩素原子；トリフルオロメチル基；トリクロロメチル基；Ｃ１～Ｃ４アルキル基；Ｃ２～Ｃ４アシルオキシ基；Ｃ２～Ｃ４アルコキシカルボニル基；Ｃ１～Ｃ３アルキル基およびＣ２～Ｃ４アルコキシカルボニル基からなる群から選択される１種または２種の基で置換されてもよいアミド基；Ｃ１～Ｃ３アルキル基およびＣ２～Ｃ４アルコキシカルボニル基からなる群から選択される１種または２種以上の基で置換されてもよいピペリジニルカルボニル基；或いはＣ１～Ｃ３アルキル基およびＣ２～Ｃ４アルコキシカルボニル基からなる群から選択される１種または２種以上の基で置換されてもよいピロリジニルカルボニル基がより好ましく、フッ素原子；塩素原子；トリフルオロメチル基；トリクロロメチル基；Ｃ１～Ｃ４アルキル基；Ｃ２～Ｃ４アルコキシカルボニル基；Ｃ１～Ｃ３アルキル基およびＣ２～Ｃ４アルコキシカルボニル基からなる群から選択される１種または２種の基で置換されてもよいアミド基；或いはＣ１～Ｃ３アルキル基およびＣ２～Ｃ４アルコキシカルボニル基からなる群から選択される１種または２種以上の基で置換されてもよいピペリジニルカルボニル基がさらに好ましく、塩素原子；トリフルオロメチル基；トリクロロメチル基；メチル基；エチル基；メトキシカルボニル基；エトキシカルボニル基；アミド基；Ｃ１～Ｃ３アルキル基から選択される１種または２種の基で置換されたアミド基；ピペリジニルカルボニル基；或いはメトキシカルボニル基およびエトキシカルボニル基から選択される１種または２種以上の基で置換されたピペリジニルカルボニル基がさらに好ましい。 When there is a plurality of R 2 s, they may be the same or different. As R2 , fluorine atom; chlorine atom; bromine atom; trifluoromethyl group; trichloromethyl group; tribromomethyl group; C1 to C4 alkyl group; C2 to C5 acyloxy group; C2 to C5 alkoxycarbonyl group; C1 to C3 An amide group optionally substituted with one or two groups selected from the group consisting of alkyl groups and C2-C4 alkoxycarbonyl groups; a piperidinylcarbonyl group which may be substituted with one or more groups; or one or more groups selected from the group consisting of a C1-C3 alkyl group and a C2-C4 alkoxycarbonyl group; An optionally substituted pyrrolidinylcarbonyl group is preferred, and includes a fluorine atom; a chlorine atom; a trifluoromethyl group; a trichloromethyl group; a C1-C4 alkyl group; a C2-C4 acyloxy group; a C2-C4 alkoxycarbonyl group; An amide group optionally substituted with one or two groups selected from the group consisting of alkyl groups and C2-C4 alkoxycarbonyl groups; a piperidinylcarbonyl group which may be substituted with one or more groups; or one or more groups selected from the group consisting of a C1-C3 alkyl group and a C2-C4 alkoxycarbonyl group; An optionally substituted pyrrolidinylcarbonyl group is more preferred, and includes a fluorine atom; a chlorine atom; a trifluoromethyl group; a trichloromethyl group; a C1-C4 alkyl group; a C2-C4 alkoxycarbonyl group; a C1-C3 alkyl group and a C2- An amide group which may be substituted with one or two groups selected from the group consisting of C4 alkoxycarbonyl groups; or one group selected from the group consisting of C1 to C3 alkyl groups and C2 to C4 alkoxycarbonyl groups; A piperidinylcarbonyl group which may be substituted with two or more types of groups is more preferred, and includes a chlorine atom; a trifluoromethyl group; a trichloromethyl group; a methyl group; an ethyl group; a methoxycarbonyl group; an ethoxycarbonyl group; an amide group; - Amido group substituted with one or two groups selected from C3 alkyl groups; piperidinylcarbonyl group; or substituted with one or more groups selected from methoxycarbonyl group and ethoxycarbonyl group More preferred are piperidinylcarbonyl groups.
　Ｒ２は、Ｒ２の結合するベンゼン環を構成する炭素原子のうち、ピペラジン環を構成する窒素原子と直接結合する炭素原子に対してオルト位に位置する２個の炭素原子のうちの１個または２個に結合していることが好ましく、１個のオルト位炭素原子に結合していることがより好ましい。Ｒ２は、パラ位に位置する炭素原子に結合してもよい。Ｒ２は、（ａ）オルト位の２個の炭素原子のうちの１個のみに結合していること、（ｂ）オルト位の２個の炭素原子のうちの１個とパラ位の炭素原子に結合していること、および（ｃ）オルト位の２個の炭素原子のうちの１個とメタ位の２個の炭素原子のうちの１個に結合していることが好ましい。Ｒ２は、（ａ）オルト位の２個の炭素原子のうちの１個のみに結合していること、または（ｂ）オルト位の２個の炭素原子のうちの１個とパラ位の炭素原子に結合していることがより好ましい。 R 2 is one of the two carbon atoms located at the ortho position to the carbon atom directly bonded to the nitrogen atom forming the piperazine ring, among the carbon atoms forming the benzene ring to which R 2 is bonded. Or preferably, it is bonded to two carbon atoms, and more preferably to one ortho carbon atom. R 2 may be bonded to a carbon atom located at the para position. R 2 is (a) bonded to only one of the two carbon atoms in the ortho position, (b) one of the two carbon atoms in the ortho position and a carbon atom in the para position. and (c) preferably one of the two carbon atoms at the ortho position and one of the two carbon atoms at the meta position. R2 is (a) bonded to only one of the two carbon atoms in the ortho position, or (b) a carbon atom in the para position to one of the two ortho carbon atoms. More preferably, it is bonded to.
　オルト位に位置する炭素原子に結合するＲ２としては、ハロゲン原子、トリハロゲノメチル基、またはＣ１～Ｃ３アルキル基が好ましく、フッ素原子、塩素原子、臭素原子、トリフルオロメチル基、メチル基、またはエチル基がより好ましく、塩素原子、トリフルオロメチル基、またはメチル基が特に好ましい。 R2 bonded to the carbon atom located at the ortho position is preferably a halogen atom, a trihalogenomethyl group, or a C1-C3 alkyl group, such as a fluorine atom, a chlorine atom, a bromine atom, a trifluoromethyl group, a methyl group, or an ethyl group. A group is more preferable, and a chlorine atom, a trifluoromethyl group, or a methyl group is particularly preferable.
　パラ位に位置する炭素原子に結合するＲ２は、例えばハロゲン原子、アルキル基；アシルオキシ基；アルコキシカルボニル基；アルキル基およびアルコキシカルボニル基からなる群から選択される１種または２種の基で置換されてもよいアミド基；アルキル基およびアルコキシカルボニル基からなる群から選択される１種または２種以上の基で置換されてもよいピペリジニルカルボニル基；或いはアルキル基およびアルコキシカルボニル基からなる群から選択される１種または２種以上の基で置換されてもよいピロリジニルカルボニル基であってよく、塩素原子；臭素原子；フッ素原子；Ｃ１～Ｃ３アルキル基；Ｃ２～Ｃ４アシルオキシ基；Ｃ２～Ｃ４アルコキシカルボニル基；Ｃ１～Ｃ３アルキル基およびＣ２～Ｃ４アルコキシカルボニル基からなる群から選択される１種または２種の基で置換されてもよいアミド基；Ｃ１～Ｃ３アルキル基およびＣ２～Ｃ４アルコキシカルボニル基からなる群から選択される１種または２種以上の基で置換されてもよいピペリジニルカルボニル基；或いはＣ１～Ｃ３アルキル基およびＣ２～Ｃ４アルコキシカルボニル基からなる群から選択される１種または２種以上の基で置換されてもよいピロリジニルカルボニル基が好ましく、塩素原子；メチル基；エチル基；メトキシカルボニル基；エトキシカルボニル基；アミド基；Ｃ１～Ｃ３アルキル基から選択される１種または２種の基で置換されたアミド基；ピペリジニルカルボニル基；或いはメトキシカルボニル基およびエトキシカルボニル基から選択される１種または２種以上の基で置換されてもよいピペリジニルカルボニル基がさらに好ましい。 R2 bonded to the carbon atom located at the para position is substituted with one or two groups selected from the group consisting of, for example, a halogen atom, an alkyl group; an acyloxy group; an alkoxycarbonyl group; an alkyl group and an alkoxycarbonyl group. an amide group which may be substituted; a piperidinylcarbonyl group which may be substituted with one or more groups selected from the group consisting of an alkyl group and an alkoxycarbonyl group; or a group consisting of an alkyl group and an alkoxycarbonyl group It may be a pyrrolidinylcarbonyl group which may be substituted with one or more groups selected from: chlorine atom; bromine atom; fluorine atom; C1-C3 alkyl group; C2-C4 acyloxy group; C2 -C4 alkoxycarbonyl group; amide group optionally substituted with one or two groups selected from the group consisting of C1-C3 alkyl group and C2-C4 alkoxycarbonyl group; C1-C3 alkyl group and C2-C4 a piperidinylcarbonyl group optionally substituted with one or more groups selected from the group consisting of alkoxycarbonyl groups; or selected from the group consisting of C1-C3 alkyl groups and C2-C4 alkoxycarbonyl groups A pyrrolidinylcarbonyl group which may be substituted with one or more groups is preferred, and is selected from a chlorine atom; a methyl group; an ethyl group; a methoxycarbonyl group; an ethoxycarbonyl group; an amide group; a C1-C3 alkyl group. amide group substituted with one or two groups; piperidinylcarbonyl group; or piperidinyl optionally substituted with one or more groups selected from methoxycarbonyl group and ethoxycarbonyl group; More preferred is a carbonyl group.
　ｎとしては、０～４の整数が好ましく、０～３の整数がより好ましく、０、１、または２がさらに好ましく、１または２がさらに好ましく、２が特に好ましい。 n is preferably an integer from 0 to 4, more preferably an integer from 0 to 3, even more preferably 0, 1, or 2, even more preferably 1 or 2, and particularly preferably 2.
　本発明において「ハロゲン原子」としては、フッ素原子、塩素原子、臭素原子およびヨウ素原子が挙げられる。好ましくはフッ素原子、塩素原子、および臭素原子、より好ましくはフッ素原子、および塩素原子が挙げられる。 In the present invention, the "halogen atom" includes a fluorine atom, a chlorine atom, a bromine atom, and an iodine atom. Preferable examples include fluorine atom, chlorine atom, and bromine atom, more preferably fluorine atom and chlorine atom.
　本発明において「アルキル基」としては、例えば、直鎖状、分枝状、または環状構造を含む、Ｃ１～Ｃ８アルキル基が挙げられ、好ましくはＣ１～Ｃ６アルキル基、より好ましくはＣ１～Ｃ４アルキル基、特に好ましくはＣ１～Ｃ３アルキル基である。具体的には、直鎖状または分枝状のアルキル基としては、メチル基、エチル基、１－プロピル基、２－プロピル基、１－ブチル基、２－ブチル基、イソブチル基、ｔｅｒｔ－ブチル基、ｎ－ペンチル基、ネオペンチル基、ｎ－ヘキシル基、イソヘキシル基、３－メチルペンチル基等が挙げられ、環状構造を含むアルキル基としては、シクロプロピル基、シクロプロピルメチル基、シクロブチル基、シクロブチルメチル基、シクロペンチル基、シクロペンチルメチル基、シクロヘキシル基、シクロヘキシルメチル基、シクロヘキシルエチル基等が挙げられる。好ましくは、メチル基、エチル基、２－プロピル基、ｔ－ブチル基、シクロプロピル基等が挙げられる。  In the present invention, the "alkyl group" includes, for example, a C1-C8 alkyl group containing a linear, branched, or cyclic structure, preferably a C1-C6 alkyl group, more preferably a C1-C4 alkyl group. groups, particularly preferably C1-C3 alkyl groups. Specifically, linear or branched alkyl groups include methyl group, ethyl group, 1-propyl group, 2-propyl group, 1-butyl group, 2-butyl group, isobutyl group, and tert-butyl group. group, n-pentyl group, neopentyl group, n-hexyl group, isohexyl group, 3-methylpentyl group, etc., and examples of alkyl groups containing a cyclic structure include cyclopropyl group, cyclopropylmethyl group, cyclobutyl group, cyclo Examples include butylmethyl group, cyclopentyl group, cyclopentylmethyl group, cyclohexyl group, cyclohexylmethyl group, cyclohexylethyl group, and the like. Preferred examples include methyl group, ethyl group, 2-propyl group, t-butyl group, and cyclopropyl group.​
　本発明において「ハロゲン原子で置換されたアルキル基」としては、例えば、ハロゲン原子で置換された、直鎖状、分枝状、または環状構造を含む、前記アルキル基が挙げられる。アルキル基が有する置換基の数は１～置換可能な最大の数であり、好ましくは３～置換可能な最大の数である。当該ハロゲン原子が複数あるときは同一でも異なってもよいが、同一であることが好ましい。例えば、置換可能な最大の数のハロゲン原子で置換されたＣ１～Ｃ４アルキル基が挙げられ、好ましくは置換可能な最大の数のハロゲン原子で置換されたＣ１～Ｃ３アルキル基、より好ましくは置換可能な最大の数のハロゲン原子で置換されたＣ１～Ｃ２アルキル基、特に好ましくはトリハロゲノメチル基である。ハロゲン原子で置換されたアルキル基の例としては、パーフルオロ－ｎ－ブチル基、パーフルオロ－ｔ－ブチル基、パーフルオロ－ｎ－プロピル基、パーフルオロ－ｉ－プロピル基、ペンチルフルオロエチル基、トリフルオロメチル基、パークロロ－ｎ－ブチル基、パークロロ－ｔ－ブチル基、パークロロ－ｎ－プロピル基、パークロロ－ｉ－プロピル基、ペンタクロロエチル基、トリクロロメチル基、パーブロモ－ｎ－ブチル基、パーブロモ－ｔ－ブチル基、パーブロモ－ｎ－プロピル基、パーブロモ－ｉ－プロピル基、ペンタブロモエチル基、トリブロモメチル基、パーヨード－ｎ－ブチル基、パーヨード－ｔ－ブチル基、パーヨード－ｎ－プロピル基、パーヨード－ｉ－プロピル基、ペンタヨードエチル基、トリヨードメチル基が挙げられる。ハロゲン原子で置換されたアルキル基の好ましい例としては、ペンチルフルオロエチル基、トリフルオロメチル基、ペンタクロロエチル基、トリクロロメチル基、ペンタブロモエチル基、トリブロモメチル基、ペンタヨードエチル基、トリヨードメチル基が挙げられ、トリフルオロメチル基、トリクロロメチル基、トリブロモメチル基がより好ましく、トリフルオロメチル基、トリクロロメチル基がより一層好ましい。 In the present invention, the "alkyl group substituted with a halogen atom" includes, for example, the alkyl group substituted with a halogen atom and having a linear, branched, or cyclic structure. The number of substituents an alkyl group has is 1 to the maximum number that can be substituted, preferably 3 to the maximum number that can be substituted. When there is a plurality of halogen atoms, they may be the same or different, but are preferably the same. Examples include C1 to C4 alkyl groups substituted with the maximum number of substitutable halogen atoms, preferably C1 to C3 alkyl groups substituted with the maximum number of substitutable halogen atoms, more preferably substitutable A C1-C2 alkyl group substituted with the maximum number of halogen atoms, particularly preferably a trihalogenomethyl group. Examples of alkyl groups substituted with halogen atoms include perfluoro-n-butyl group, perfluoro-t-butyl group, perfluoro-n-propyl group, perfluoro-i-propyl group, pentylfluoroethyl group, Trifluoromethyl group, perchloro-n-butyl group, perchloro-t-butyl group, perchloro-n-propyl group, perchloro-i-propyl group, pentachloroethyl group, trichloromethyl group, perbromo-n-butyl group, perbromo -t-butyl group, perbromo-n-propyl group, perbromo-i-propyl group, pentabromoethyl group, tribromomethyl group, periodo-n-butyl group, periodo-t-butyl group, periodo-n-propyl group , periodo-i-propyl group, pentaiodoethyl group, and triiodomethyl group. Preferred examples of the alkyl group substituted with a halogen atom include a pentylfluoroethyl group, a trifluoromethyl group, a pentachloroethyl group, a trichloromethyl group, a pentabromoethyl group, a tribromomethyl group, a pentayodoethyl group, and a triiodoethyl group. A methyl group is mentioned, a trifluoromethyl group, a trichloromethyl group, and a tribromomethyl group are more preferable, and a trifluoromethyl group and a trichloromethyl group are even more preferable.
　本発明において「アルコキシ基」としては、例えば、直鎖状、分枝状、または環状構造を含む、Ｃ１～Ｃ８アルコキシ基が挙げられ、好ましくはＣ１～Ｃ６アルコキシ基、より好ましくはＣ１～Ｃ４アルコキシ基、特に好ましくはＣ１～Ｃ３アルコキシ基である。具体的には、直鎖状または分枝状のアルコキシ基としては、メトキシ基、エトキシ基、１－プロポキシ基、２－プロポキシ基、１－ブトキシ基、２－ブトキシ基、イソブトキシ基、ｔｅｒｔ－ブトキシ基、ｎ－ペンチルオキシ基、ネオペンチルオキシ基、ｎ－ヘキシルオキシ基、イソヘキシルオキシ基、３－メチルペンチルオキシ基等が挙げられる。環状構造を含むアルコキシ基としては、シクロプロポキシ基、シクロプロピルメトキシ基、シクロブチロキシ基、シクロブチルメトキシ基、シクロペンチロキシ基、シクロペンチルメトキシ基、シクロヘキシロキシ基、シクロヘキシルメトキシ基、シクロヘキシルエトキシ基等が挙げられる。好ましくは、メトキシ基、エトキシ基、２－プロポキシ基、ｔｅｒｔ－ブトキシ基、シクロプロポキシ基等が挙げられる。 In the present invention, the "alkoxy group" includes, for example, a C1-C8 alkoxy group containing a linear, branched, or cyclic structure, preferably a C1-C6 alkoxy group, more preferably a C1-C4 alkoxy group. groups, particularly preferably C1-C3 alkoxy groups. Specifically, linear or branched alkoxy groups include methoxy group, ethoxy group, 1-propoxy group, 2-propoxy group, 1-butoxy group, 2-butoxy group, isobutoxy group, and tert-butoxy group. group, n-pentyloxy group, neopentyloxy group, n-hexyloxy group, isohexyloxy group, 3-methylpentyloxy group and the like. Examples of alkoxy groups containing a cyclic structure include cyclopropoxy group, cyclopropylmethoxy group, cyclobutyroxy group, cyclobutylmethoxy group, cyclopentyloxy group, cyclopentylmethoxy group, cyclohexyloxy group, cyclohexylmethoxy group, cyclohexylethoxy group, etc. Can be mentioned. Preferred examples include methoxy group, ethoxy group, 2-propoxy group, tert-butoxy group, and cyclopropoxy group.
　本発明において「アシルオキシ基」としては、例えば、直鎖状、分枝状、または環状構造を含む、Ｃ２～Ｃ９アシルオキシ基が挙げられ、好ましくはＣ２～Ｃ７アシルオキシ基、より好ましくはＣ２～Ｃ５アシルオキシ基、特に好ましくはＣ２～Ｃ４アシルオキシ基である。具体的には、直鎖状または分枝状のアシルオキシ基としては、メトキシカルボニル基、エトキシカルボニル基、１－プロポキシカルボニル基、２－プロポキシカルボニル基、１－ブトキシカルボニル基、２－ブトキシカルボニル基、イソブトキシカルボニル基、ｔｅｒｔ－ブトキシカルボニル基、ｎ－ペンチルオキシカルボニル基、ネオペンチルオキシカルボニル基、ｎ－ヘキシルオキシカルボニル基、イソヘキシルオキシカルボニル基、３－メチルペンチルオキシカルボニル基等が挙げられる。環状構造を含むアシルオキシ基としては、シクロプロポキシカルボニル基、シクロプロピルメトキシカルボニル基、シクロブチロキシカルボニル基、シクロブチルメトキシカルボニル基、シクロペンチロキシカルボニル基、シクロペンチルメトキシカルボニル基、シクロヘキシロキシカルボニル基、シクロヘキシルメトキシカルボニル基、シクロヘキシルエトキシカルボニル基等が挙げられる。好ましくは、メトキシカルボニル基、エトキシカルボニル基、２－プロポキシカルボニル基、ｔｅｒｔ－ブトキシカルボニル基、シクロプロポキシカルボニル基等が挙げられる。 In the present invention, the "acyloxy group" includes, for example, a C2-C9 acyloxy group containing a linear, branched, or cyclic structure, preferably a C2-C7 acyloxy group, more preferably a C2-C5 acyloxy group. groups, particularly preferably C2-C4 acyloxy groups. Specifically, the linear or branched acyloxy group includes a methoxycarbonyl group, an ethoxycarbonyl group, a 1-propoxycarbonyl group, a 2-propoxycarbonyl group, a 1-butoxycarbonyl group, a 2-butoxycarbonyl group, Examples include isobutoxycarbonyl group, tert-butoxycarbonyl group, n-pentyloxycarbonyl group, neopentyloxycarbonyl group, n-hexyloxycarbonyl group, isohexyloxycarbonyl group, and 3-methylpentyloxycarbonyl group. Acyloxy groups containing a cyclic structure include cyclopropoxycarbonyl group, cyclopropylmethoxycarbonyl group, cyclobutyroxycarbonyl group, cyclobutylmethoxycarbonyl group, cyclopentyloxycarbonyl group, cyclopentylmethoxycarbonyl group, cyclohexyloxycarbonyl group, cyclohexylmethoxy Examples include carbonyl group, cyclohexylethoxycarbonyl group, and the like. Preferred examples include methoxycarbonyl group, ethoxycarbonyl group, 2-propoxycarbonyl group, tert-butoxycarbonyl group, and cyclopropoxycarbonyl group.
　本発明において「アルコキシカルボニル基」としては、前記アルコキシ基と結合したカルボニル基が挙げられ、例えば直鎖状、分枝状、または環状構造を含む、Ｃ１～Ｃ８アルコキシ基と結合したカルボニル基が挙げられ、好ましくはＣ２～Ｃ７アルコキシカルボニル基、より好ましくはＣ２～Ｃ５アルコキシカルボニル基、特に好ましくはＣ２～Ｃ４アルコキシカルボニル基である。アルコキシカルボニル基を構成する直鎖状または分枝状のアルコキシ基としては、メトキシ基、エトキシ基、１－プロポキシ基、２－プロポキシ基、１－ブトキシ基、２－ブトキシ基、イソブトキシ基、ｔｅｒｔ－ブトキシ基、ｎ－ペンチルオキシ基、ネオペンチルオキシ基、ｎ－ヘキシルオキシ基、イソヘキシルオキシ基、３－メチルペンチルオキシ基等が挙げられる。アルコキシカルボニル基を構成する環状構造を含むアルコキシ基としては、シクロプロポキシ基、シクロプロピルメトキシ基、シクロブチロキシ基、シクロブチルメトキシ基、シクロペンチロキシ基、シクロペンチルメトキシ基、シクロヘキシロキシ基、シクロヘキシルメトキシ基、シクロヘキシルエトキシ基等が挙げられる。アルコキシカルボニル基を構成するアルコキシ基としては、好ましくは、メトキシ基、エトキシ基、２－プロポキシ基、ｔｅｒｔ－ブトキシ基、シクロプロポキシ基等が挙げられる。 In the present invention, the "alkoxycarbonyl group" includes a carbonyl group bonded to the alkoxy group, such as a carbonyl group bonded to a C1 to C8 alkoxy group, which has a linear, branched, or cyclic structure. and is preferably a C2-C7 alkoxycarbonyl group, more preferably a C2-C5 alkoxycarbonyl group, particularly preferably a C2-C4 alkoxycarbonyl group. Examples of linear or branched alkoxy groups constituting the alkoxycarbonyl group include methoxy group, ethoxy group, 1-propoxy group, 2-propoxy group, 1-butoxy group, 2-butoxy group, isobutoxy group, tert- Examples include butoxy group, n-pentyloxy group, neopentyloxy group, n-hexyloxy group, isohexyloxy group, and 3-methylpentyloxy group. Examples of alkoxy groups containing a cyclic structure constituting an alkoxycarbonyl group include a cyclopropoxy group, a cyclopropylmethoxy group, a cyclobutyroxy group, a cyclobutylmethoxy group, a cyclopentyloxy group, a cyclopentylmethoxy group, a cyclohexyloxy group, and a cyclohexylmethoxy group. , cyclohexylethoxy group, and the like. Preferred examples of the alkoxy group constituting the alkoxycarbonyl group include methoxy group, ethoxy group, 2-propoxy group, tert-butoxy group, and cyclopropoxy group.
　本発明において「アルキル基およびアルコキシカルボニル基からなる群から選択される１種または２種の基で置換されてもよいアミド基」は、アミド基、モノまたはジ－アルキルアミド基、モノまたはジ－アルコキシカルボニルアミド基、アルキル基およびアルコキシカルボニル基でジ置換されたアミド基を包含し、好ましくはアミド基、ジアルキルアミド基、より好ましくはジアルキルアミド基である。アミド基が２つの基で置換される場合、置換基は同一でも異なってもよい。モノアルキルアミド基としては、前記アルキル基でモノ置換されたアミド基が挙げられ、好ましくはＣ１～Ｃ４アルキル基でモノ置換されたアミド基、より好ましくはＣ１～Ｃ３アルキル基でモノ置換されたアミド基が挙げられ、特に好ましくはメチルアミド基、エチルアミド基が挙げられる。ジアルキルアミド基としては、前記アルキル基でジ置換されたアミド基が挙げられ、好ましくはＣ１～Ｃ４アルキル基でジ置換されたアミド基、より好ましくはＣ１～Ｃ３アルキル基でジ置換されたアミド基、特に好ましくはジメチルアミド基、ジエチルアミド基、エチルメチルアミド基が挙げられる。モノアルコキシカルボニルアミド基としては、前記アルコキシカルボニル基でモノ置換されたアミド基が挙げられ、好ましくはＣ２～Ｃ５アルコキシカルボニル基でモノ置換されたアミド基、より好ましくはＣ２～Ｃ４アルコキシカルボニル基でモノ置換されたアミド基、特に好ましくはメトキシカルボニルアミド、エトキシカルボニルアミド基が挙げられる。ジアルコキシカルボニルアミド基としては、前記アルコキシカルボニル基でジ置換されたアミド基が挙げられ、好ましくはＣ２～Ｃ５アルコキシカルボニル基でジ置換されたアミド基、より好ましくはＣ２～Ｃ４アルコキシカルボニル基でジ置換されたアミド基、特に好ましくはジ（メトキシカルボニル）アミド、ジ（エトキシカルボニル）アミド基が挙げられる。アルキル基およびアルコキシカルボニル基で置換されたアミド基としては、前記アルキル基と前記アルコキシカルボニル基でジ置換されたアミド基が挙げられ、好ましくはＣ１～Ｃ４アルキル基とＣ２～Ｃ５アルコキシカルボニル基でジ置換されたアミド基、より好ましくはＣ１～Ｃ３アルキル基とＣ２～Ｃ４アルコキシカルボニル基でジ置換されたアミド基、特に好ましくはメチルメトキシカルボニルアミド基、メチルエトキシカルボニルアミド基、エチルメトキシカルボニルアミド基、エチルエトキシカルボニルアミド基が挙げられる。 In the present invention, "an amide group which may be substituted with one or two groups selected from the group consisting of an alkyl group and an alkoxycarbonyl group" means an amide group, a mono- or di-alkyl amide group, a mono- or di- It includes an alkoxycarbonylamide group, an alkyl group, and an amide group di-substituted with an alkoxycarbonyl group, preferably an amide group or a dialkylamide group, and more preferably a dialkylamide group. When an amide group is substituted with two groups, the substituents may be the same or different. Examples of the monoalkyl amide group include amide groups mono-substituted with the above-mentioned alkyl groups, preferably amide groups mono-substituted with C1-C4 alkyl groups, more preferably amide groups mono-substituted with C1-C3 alkyl groups. Among these groups, particularly preferred are methylamide and ethylamide groups. Examples of dialkylamide groups include amide groups di-substituted with the above-mentioned alkyl groups, preferably amide groups di-substituted with C1-C4 alkyl groups, more preferably amide groups di-substituted with C1-C3 alkyl groups. Particularly preferred are dimethylamide group, diethylamide group, and ethylmethylamide group. Examples of the monoalkoxycarbonylamide group include an amide group mono-substituted with the above-mentioned alkoxycarbonyl group, preferably an amide group mono-substituted with a C2-C5 alkoxycarbonyl group, more preferably a mono-substituted amide group with a C2-C4 alkoxycarbonyl group. Substituted amide groups, particularly preferred are methoxycarbonylamide and ethoxycarbonylamide groups. The dialkoxycarbonylamide group includes an amide group di-substituted with the alkoxycarbonyl group described above, preferably an amide group di-substituted with a C2-C5 alkoxycarbonyl group, more preferably a di-substituted amide group with a C2-C4 alkoxycarbonyl group. Substituted amide groups, particularly preferred are di(methoxycarbonyl)amide and di(ethoxycarbonyl)amide groups. The amide group substituted with an alkyl group and an alkoxycarbonyl group includes an amide group di-substituted with the alkyl group and the alkoxycarbonyl group, preferably a di-substituted amide group with a C1-C4 alkyl group and a C2-C5 alkoxycarbonyl group. A substituted amide group, more preferably a di-substituted amide group with a C1 to C3 alkyl group and a C2 to C4 alkoxycarbonyl group, particularly preferably a methylmethoxycarbonylamide group, a methylethoxycarbonylamide group, an ethylmethoxycarbonylamide group, Examples include ethyl ethoxycarbonylamide group.
　本発明において「アルキル基およびアルコキシカルボニル基からなる群から選択される１種または２種以上の基で置換されてもよいピペリジニルカルボニル基」においてピペリジニルカルボニル基が有する置換基の数は０～５であり、好ましくは０～４、より好ましくは０～３、より一層好ましくは０～２、特に好ましくは０または１である。当該置換基が複数あるときは同一でも異なってもよい。ピペリジニルカルボニル基が置換基を有する場合、少なくとも１個の置換基がピペリジン環の３位（詳細には、ピペリジン環を構成する窒素原子を１位とした場合の３位）の炭素原子に結合していることが好ましい。置換基としては、例えば前記のアルキル基および前記のアルコキシカルボニル基が挙げられ、好ましくはＣ１～Ｃ４アルキル基、Ｃ２～Ｃ５アルコキシカルボニル基、より好ましくはＣ１～Ｃ３アルキル基、Ｃ２～Ｃ４アルコキシカルボニル基、特に好ましくはメチル基、エチル基、メトキシカルボニル基、エトキシカルボニル基が挙げられる。 In the present invention, in the "piperidinylcarbonyl group which may be substituted with one or more groups selected from the group consisting of alkyl groups and alkoxycarbonyl groups", the number of substituents that the piperidinylcarbonyl group has is The number is 0 to 5, preferably 0 to 4, more preferably 0 to 3, even more preferably 0 to 2, particularly preferably 0 or 1. When there are multiple substituents, they may be the same or different. When the piperidinylcarbonyl group has a substituent, at least one substituent is attached to the carbon atom at the 3-position of the piperidine ring (specifically, the 3-position when the nitrogen atom constituting the piperidine ring is the 1-position). Preferably, they are bonded. Examples of the substituent include the above-mentioned alkyl group and the above-mentioned alkoxycarbonyl group, preferably a C1-C4 alkyl group, a C2-C5 alkoxycarbonyl group, more preferably a C1-C3 alkyl group, a C2-C4 alkoxycarbonyl group. Particularly preferred are a methyl group, an ethyl group, a methoxycarbonyl group, and an ethoxycarbonyl group.
　本発明において「アルキル基およびアルコキシカルボニル基からなる群から選択される１種または２種以上の基で置換されてもよいピロリジニルカルボニル基」においてピロリジニルカルボニル基が有する置換基の数は０～４であり、好ましくは０～３、より好ましくは０～２、特に好ましくは０または１である。当該置換基が複数あるときは同一でも異なってもよい。ピロリジニルカルボニル基が置換基を有する場合、少なくとも１個の置換基がピロリジン環の３位（詳細には、ピロリジン環を構成する窒素原子を１位とした場合の３位）の炭素原子に結合していることが好ましい。置換基としては、例えば前記のアルキル基および前記のアルコキシカルボニル基が挙げられ、好ましくはＣ１～Ｃ４アルキル基、Ｃ２～Ｃ５アルコキシカルボニル基、より好ましくはＣ１～Ｃ３アルキル基、Ｃ２～Ｃ４アルコキシカルボニル基、特に好ましくはメチル基、エチル基、メトキシカルボニル基、エトキシカルボニル基が挙げられる。 In the present invention, in the "pyrrolidinylcarbonyl group which may be substituted with one or more groups selected from the group consisting of alkyl groups and alkoxycarbonyl groups", the number of substituents that the pyrrolidinylcarbonyl group has is The number is 0 to 4, preferably 0 to 3, more preferably 0 to 2, particularly preferably 0 or 1. When there are multiple substituents, they may be the same or different. When the pyrrolidinylcarbonyl group has a substituent, at least one substituent is attached to the carbon atom at the 3-position of the pyrrolidine ring (specifically, the 3-position when the nitrogen atom constituting the pyrrolidine ring is the 1-position). Preferably, they are bonded. Examples of the substituent include the above-mentioned alkyl group and the above-mentioned alkoxycarbonyl group, preferably a C1-C4 alkyl group, a C2-C5 alkoxycarbonyl group, more preferably a C1-C3 alkyl group, a C2-C4 alkoxycarbonyl group. Particularly preferred are a methyl group, an ethyl group, a methoxycarbonyl group, and an ethoxycarbonyl group.
　本発明において、「予防」とは、心不全または骨格筋不全の発症を事前に防ぐことを意味する。 In the present invention, "prevention" means preventing the onset of heart failure or skeletal muscle failure in advance.
　本発明において、「処置」とは、 (１) 心不全または骨格筋不全の発症を遅延させる； (２) 心不全または骨格筋不全の症状の進行、増悪または悪化を減速または停止させる； (３) 心不全または骨格筋不全の症状の寛解をもたらす；あるいは (４) 心不全または骨格筋不全を治癒させることを目的とする方法またはプロセスを意味する。処置は、予防的措置として疾患または状態の発症前に施してもよいし、あるいはまた、処置は、疾患の発症後に施すことができる。 In the present invention, "treatment" means: (1) Delaying the onset of heart failure or skeletal muscle failure; (2) Slowing down or stopping the progression, exacerbation, or deterioration of symptoms of heart failure or skeletal muscle failure; (3) Heart failure or (4) means a method or process aimed at curing heart failure or skeletal muscle failure. Treatment may be administered before the onset of the disease or condition as a prophylactic measure, or alternatively, treatment can be administered after the onset of the disease.
　本発明において、「心不全」とは、代謝組織が必要とする速度で、心臓が血液を送り出さない心臓機能の異常を意味する。心不全は、鬱血性心不全、心筋梗塞、不整頻拍、家族性肥大型心筋症、虚血性心疾患、突発性拡張型心筋症、心筋炎等の広範囲の疾患状態を含む。心不全は、限定されるわけではないが、虚血性、先天性、リューマチ性、ウイルス性、毒性または特発性の形態を含む多くの要因から起こり得る。心不全に含まれる慢性心肥大は、鬱血性心不全および心停止の前兆となる重大な疾患状態である。本発明は、慢性心肥大を予防および／または処置するための医薬組成物も包含する。 In the present invention, "heart failure" refers to an abnormality in heart function in which the heart does not pump blood at the rate required by metabolic tissues. Heart failure includes a wide range of disease states such as congestive heart failure, myocardial infarction, tachycardia, familial hypertrophic cardiomyopathy, ischemic heart disease, idiopathic dilated cardiomyopathy, and myocarditis. Heart failure can result from many causes including, but not limited to, ischemic, congenital, rheumatic, viral, toxic or idiopathic forms. Chronic cardiac hypertrophy, a component of heart failure, is a serious disease state that is a precursor to congestive heart failure and cardiac arrest. The present invention also encompasses pharmaceutical compositions for preventing and/or treating chronic cardiac hypertrophy.
　本発明において、「骨格筋不全」とは、筋肉がその仕事を遂行する能力がない状態を意味し、筋力の低下または持久力の低下、またはその両者として症状が現れる。これには、筋ジストロフィーが含まれる。 In the present invention, "skeletal muscle insufficiency" refers to a state in which muscles do not have the ability to perform their work, and symptoms appear as a decrease in muscle strength, a decrease in endurance, or both. This includes muscular dystrophy.
　本発明において医薬組成物とは、通常、疾患の予防もしくは処置、または検査・診断のための薬剤を意味する。 In the present invention, a pharmaceutical composition usually means a drug for preventing or treating a disease, or for testing or diagnosing a disease.
　特定の理論にとらわれるものでないが、本発明におけるTRPC3/6/7チャネル活性化物質は、血圧を下げず、また急性増悪に対して効能を示す。他方、TRPC3/6/7チャネル阻害物質では、急性増悪に対する効果は知られていない。そこで、本発明では、心不全または骨格筋不全における急性期初期の段階では、本発明のTRPC3/6/7チャネル活性化物質を投与し、次いでTRPC3/6/7チャネル阻害物質を投与する治療方式が、理想的だと考えられる。 Without being bound by any particular theory, the TRPC3/6/7 channel activator of the present invention does not lower blood pressure and exhibits efficacy against acute exacerbation. On the other hand, TRPC3/6/7 channel inhibitors are not known to have any effect on acute exacerbation. Therefore, the present invention provides a treatment method in which the TRPC3/6/7 channel activator of the present invention is administered and then the TRPC3/6/7 channel inhibitor is administered in the early acute stage of heart failure or skeletal muscle failure. , is considered ideal.
　本発明の医薬組成物は、当業者に公知の方法で製剤化することが可能である。本発明の医薬組成物は、非経口投与および経口投与のいずれによっても投与することができる。非経口投与の場合、例えば、注射剤型、経鼻投与剤型、経肺投与剤型、経皮投与型の組成物とすることができる。例えば、静脈内注射、筋肉内注射、腹腔内注射、皮下注射などにより全身または局部的に投与することができる。経口投与の場合、錠剤、カプセル剤、丸剤、顆粒剤、散剤、シロップ剤等の組成物とすることができる。 The pharmaceutical composition of the present invention can be formulated by methods known to those skilled in the art. The pharmaceutical composition of the present invention can be administered either parenterally or orally. In the case of parenteral administration, the composition can be, for example, in the form of an injection, a nasal administration, a pulmonary administration, or a transdermal administration. For example, it can be administered systemically or locally by intravenous injection, intramuscular injection, intraperitoneal injection, subcutaneous injection, etc. For oral administration, the composition may be a tablet, capsule, pill, granule, powder, syrup, or the like.
　投与方法は、患者の年齢、症状により適宜選択することができる。ポリペプチドを含有する医薬組成物の投与量は、例えば、１回につき体重1kgあたり0.0001mgから1000mgの範囲に設定することが可能である。または、例えば、患者あたり0.001～100000mgの投与量とすることもできるが、本発明はこれらの数値に必ずしも制限されるものではない。投与量および投与方法は、患者の体重、年齢、症状などにより変動するが、当業者であればそれらの条件を考慮し適当な投与量および投与方法を設定することが可能である。 The administration method can be selected as appropriate depending on the age and symptoms of the patient. The dosage of the pharmaceutical composition containing the polypeptide can be set, for example, in the range of 0.0001 mg to 1000 mg per kg of body weight per dose. Alternatively, the dosage may be, for example, 0.001 to 100000 mg per patient, although the present invention is not necessarily limited to these values. The dosage and method of administration vary depending on the patient's weight, age, symptoms, etc., and those skilled in the art can take these conditions into account and set an appropriate dosage and method of administration.
　本発明は別の態様として、心不全または骨格筋不全を処置もしくは予防するための方法であって、TRPC3/6/7チャネル活性化物質を、そのような処置等を必要としている対象に投与することを含む方法、好ましくは、本発明の化合物をそのような対象に投与することを含む方法に関する。
　さらに、本発明は別の態様として、心不全または骨格筋不全を処置もしくは予防するためのTRPC3/6/7チャネル活性化物質に関する。
　本発明はさらなる別の態様として、心不全または骨格筋不全を処置もしくは予防するための医薬を製造するための、TRPC3/6/7チャネル活性化物質の使用に関する。 Another aspect of the present invention is a method for treating or preventing heart failure or skeletal muscle failure, which comprises administering a TRPC3/6/7 channel activator to a subject in need of such treatment. , preferably administering a compound of the invention to such a subject.
Furthermore, in another aspect, the present invention relates to TRPC3/6/7 channel activators for treating or preventing heart failure or skeletal muscle failure.
In yet another aspect, the present invention relates to the use of a TRPC3/6/7 channel activator for the manufacture of a medicament for treating or preventing heart failure or skeletal muscle failure.
　本発明は別の形態において、　式（１）：
Figure JPOXMLDOC01-appb-C000017
 ［式中、
　Ａは、炭素原子または窒素原子を示し、
　Ｂは、炭素原子または窒素原子を示し、
　Ｒ１は、ハロゲン原子、アルキル基、アルコキシ基、アシルオキシ基、アルコキシカルボニル基、またはハロゲン原子で置換されたアルキル基を示し、複数あるときは同一または異なってよく、
　ｍは、０～５の整数を示し、
　ｎは、２～５の整数を示し、
　複数あるＲ２のうち、少なくとも１つはハロゲン原子、アルキル基またはトリフルオロメチル基であり；残りのＲ２は、１）単数または複数のハロゲン原子で置換され、かつアルキル基およびアルコキシカルボニル基からなる群から選択される１種または２種以上の基で置換されてもよいピペリジニルカルボニル基；或いは２）単数または複数のハロゲン原子で置換され、かつアルキル基およびアルコキシカルボニル基からなる群から選択される１種または２種以上の基で置換されてもよいピロリジニルカルボニル基を示し、複数あるときは同一または異なってよく、
　ｐは、０または１を示し、そして
　ｑは、０または１を示す。］
で表される化合物に関する。本発明の化合物は、当業者に周知の製薬的に許容できる塩を形成することができ、また、当業者に周知の方法および上記文献に記載の方法により調製することができる。 In another form of the present invention, formula (1):
Figure JPOXMLDOC01-appb-C000017
 [In the formula,
A represents a carbon atom or a nitrogen atom,
B represents a carbon atom or a nitrogen atom,
R 1 represents a halogen atom, an alkyl group, an alkoxy group, an acyloxy group, an alkoxycarbonyl group, or an alkyl group substituted with a halogen atom, and when there is more than one, they may be the same or different;
m represents an integer from 0 to 5,
n represents an integer from 2 to 5,
At least one of the plurality of R 2 is a halogen atom, an alkyl group, or a trifluoromethyl group; the remaining R 2 are 1) substituted with one or more halogen atoms, and are selected from an alkyl group and an alkoxycarbonyl group. or 2) a piperidinylcarbonyl group optionally substituted with one or more groups selected from the group consisting of alkyl groups and alkoxycarbonyl groups; or 2) substituted with one or more halogen atoms; Indicates a pyrrolidinylcarbonyl group that may be substituted with one or more selected groups, and when there are multiple groups, they may be the same or different,
p represents 0 or 1, and q represents 0 or 1. ]
Regarding the compound represented by. The compounds of the invention can form pharmaceutically acceptable salts well known to those skilled in the art and can be prepared by methods well known to those skilled in the art and described in the above-mentioned literature.
＜スクリーニング方法＞
　本発明は別の形態において、心不全または骨格筋不全を予防および／または処置するための物質をスクリーニングする方法であって、TRPC3/6/7チャネル活性化物質を選別する、該スクリーニング方法を提供する。具体的には、本発明スクリーニング方法は、被験物質の存在下で、細胞中の亜鉛濃度を測定する工程と、前記亜鉛濃度が、前記被験物質の非存在下における亜鉛濃度と比較して増加していた場合に、前記被験物質はTRPC3/6/7チャネル活性化をもたらす候補物質であると判断する工程とを備える、TRPC3/6/7チャネル活性化物質の候補物質のスクリーニング方法を提供する。 <Screening method>
In another aspect, the present invention provides a method of screening for substances for preventing and/or treating heart failure or skeletal muscle failure, the screening method selecting for TRPC3/6/7 channel activators. . Specifically, the screening method of the present invention includes a step of measuring zinc concentration in cells in the presence of a test substance, and a step in which the zinc concentration increases compared to the zinc concentration in the absence of the test substance. Provided is a method for screening a candidate substance for a TRPC3/6/7 channel activating substance, comprising the step of determining that the test substance is a candidate substance that causes activation of a TRPC3/6/7 channel.
　細胞は、培養細胞である限り制限されない。例えば、マウスまたはラットの培養心筋細胞を利用する場合、心臓から単離して初代培養することが好ましい。市販の細胞株としては、HEK293細胞を使用し、TRPC3/6/7チャネルを過剰発現させることで活性を見ることができる。細胞は好ましくは、HEK293細胞であり、より好ましくはマウスまたはラットの培養心筋細胞である。 The cells are not limited as long as they are cultured cells. For example, when using cultured mouse or rat cardiomyocytes, it is preferable to isolate them from the heart and perform primary culture. HEK293 cells are used as a commercially available cell line, and activity can be observed by overexpressing TRPC3/6/7 channels. The cells are preferably HEK293 cells, more preferably cultured mouse or rat cardiomyocytes.
　被験物質とは、TRPC3/6/7チャネル活性化物質の候補物質であるか否かを評価する対象となりうる物質であり、特に制限されない。例えば、化合物、タンパク質、ペプチド、核酸、脂質、糖質、糖脂質、糖タンパク、金属等を挙げることができる。被験物質の投与方法も特に制限されない。 A test substance is a substance that can be evaluated to determine whether it is a candidate substance for a TRPC3/6/7 channel activator, and is not particularly limited. Examples include compounds, proteins, peptides, nucleic acids, lipids, carbohydrates, glycolipids, glycoproteins, metals, and the like. The method of administering the test substance is also not particularly limited.
　以下、本発明を参考例、試験例、調製例および実施例により、詳細に説明するが、これらは本発明の範囲を限定するものでなく、単なる例示であることに留意すべきである。 Hereinafter, the present invention will be explained in detail by reference examples, test examples, preparation examples, and examples, but it should be noted that these do not limit the scope of the present invention and are merely illustrative.
参考例１
血管拡張薬ヒドララジン（Hyd）静脈内投与による一過的な血圧低下に対する代償性心機能亢進
　野生型（WT）および米国NIEHS研究所から入手したTRPC6遺伝子欠損マウス（TRPC6(-/-)）（Dietrich A, Mederos YSM, Gollasch M, Gross V, Storch U, Dubrovska G, et al. (2005). Increased vascular smooth muscle contractility in TRPC6-/- mice. Molecular and cellular biology 25(16): 6980-6989. doi: 10.1128/MCB.25.16.6980-6989.2005）に血管拡張薬ヒドララジン（Hyd）を静脈内投与し、血圧低下に対する代償性心機能亢進を調べた。結果を図１に示す。図１は、野生型（WT）と比べ、TRPC6遺伝子欠損マウスでは、左心室収縮力の増加分（ΔLVdP/dtmax）が著しく減弱し、一方、心拍数の増加分（ΔHeart Rate）に差は無いことを示している。これは、TRPC6欠損マウスでは、野生型マウスと比べ、ヒドララジン誘発性の圧受容反射応答（陽性変時作用（心拍数の増加）と陽性変力作用（収縮力の増加））のうち、陽性変力作用が著しく減弱していることを意味する。 Reference example 1
Compensatory cardiac hyperactivity in response to a transient decrease in blood pressure due to intravenous administration of the vasodilator hydralazine (Hyd) Wild type (WT) and TRPC6 gene-deficient mice (TRPC6(-/-) obtained from the NIEHS Institute in the United States) (Dietrich A, Mederos YSM, Gollasch M, Gross V, Storch U, Dubrovska G, et al. (2005). Increased vascular smooth muscle contractility in TRPC6-/- mice. Molecular and cellular biology 25(16): 6980-6989. doi : 10.1128/MCB.25.16.6980-6989.2005), the vasodilator hydralazine (Hyd) was administered intravenously to examine compensatory cardiac hyperactivity in response to a drop in blood pressure. The results are shown in Figure 1. Figure 1 shows that compared to wild type (WT) mice, the increase in left ventricular contractile force (ΔLVdP/dtmax) is significantly attenuated in TRPC6 gene-deficient mice, while there is no difference in the increase in heart rate (ΔHeart Rate). It is shown that. This indicates that TRPC6-deficient mice showed a positive change in hydralazine-induced baroreflex responses (positive chronotropy (increase in heart rate) and positive inotropy (increase in contractile force)) compared to wild-type mice. This means that the force action is significantly weakened.
　以下の試験例により、TRPC6チャネル活性化が慢性心不全に対して有効かどうかの検証を行った。TRPC3/6/7チャネル活性を増強するピペラジン誘導体（PPZ）（Sawamura S, Hatano M, Takada Y, Hino K, Kawamura T, Tanikawa J, Nakagawa H, Hase H, Nakao A, Hirano M, Rotrattanadumrong R, Kiyonaka S, Mori MX, Nishida M, Hu Y, Inoue R, Nagata R, Mori Y. Screening of Transient Receptor Potential Canonical Channel Activators Identifies Novel Neurotrophic Piperazine Compounds. Mol Pharmacol. 89(3), 348-363 (2016). doi: 10.1124/mol.115.102863.）を用い、TRPC6チャネル活性化が慢性心不全に対して有効かどうか検証した。PPZ誘導体の一つであるPPZ2 (2.5mg/kg/day)を含む浸透圧ポンプを下記心不全モデルマウスの腹腔内に埋込み、その後の心機能変化を、心エコー装置を用いて経時的・非侵襲的に計測した。また、エンドポイントにおける左心室組織の心筋細胞面積はWheet Germ Agglitin (WGA)染色画像を用いて、間接の線維化はピクロシリウスレッド（picrosirius red）染色画像を用いて解析した。 The following test examples were used to verify whether TRPC6 channel activation is effective against chronic heart failure. Piperazine derivative (PPZ) that enhances TRPC3/6/7 channel activity (Sawamura S, Hatano M, Takada Y, Hino K, Kawamura T, Tanikawa J, Nakagawa H, Hase H, Nakao A, Hirano M, Rotrattanadumrong R, Kiyonaka S, MORI MX, Nishida M, Hu Y, INOUE R, NAGATA R, MORI Y. Screening of Transmential Potential Canonel Channel Activators Identification Identifies Neurotro. PHIC PIPERAZINE COMPOUNDS. Mol Pharmacol. 89 (3), 348-363 (2016). Doi : 10.1124/mol.115.102863.) to verify whether TRPC6 channel activation is effective against chronic heart failure. An osmotic pump containing PPZ2 (2.5 mg/kg/day), one of the PPZ derivatives, was implanted into the abdominal cavity of the following heart failure model mouse, and subsequent changes in cardiac function were monitored over time and non-invasively using an echocardiogram device. It was measured. In addition, the area of myocardial cells in the left ventricular tissue at the end point was analyzed using Wheat Germ Agglitin (WGA) stained images, and indirect fibrosis was analyzed using picrosirius red stained images.
　試験例にて使用しているTRPC3/6/7チャネル活性化物質PPZ2、パーフォリンおよびGSK1702934Aは以下の式で示される。
Figure JPOXMLDOC01-appb-C000018
 The TRPC3/6/7 channel activator PPZ2, perforin, and GSK1702934A used in the test example are represented by the following formula.
Figure JPOXMLDOC01-appb-C000018
試験例１
MLP欠損マウスにおけるTRPC6チャネル活性化 Test example 1
TRPC6 channel activation in MLP-deficient mice
　拡張型心筋症モデルマウスであるmuscle LIM protein (MLP)欠損マウス（Knoll R, Hoshijima M, Hoffman HM, Person V, Lorenzen-Schmidt I, Bang ML, Hayashi T, Shiga N, Yasukawa H, Schaper W, McKenna W, Yokoyama M, Schork NJ, Omens JH, McCulloch AD, Kimura A, Gregorio CC, Poller W, Schaper J, Schultheiss HP and Chien KR The cardiac mechanical stretch sensor machinery involves a Z disc complex that is defective in a subset of human dilated cardiomyopathy. Cell 111 (2002b), pp. 943-55.）は、出生5週後から心不全を発症する。8～9週令の雄性マウスにPPZ2含有浸透圧ポンプを埋め込んだ後の心機能（駆出率（ejection fraction (EF)）と左室内径短縮率fractional shortening (FS)）を計測した結果、溶媒（50% (v/v) ジエチルスルホキシド (DMSO)/ 50% (v/v) ポリエチレングリコール300）のみを投与したMLP欠損マウスと比べて、１～３週間後の心機能が有意に高くなることがわかった（図２）。PPZ2投与3週間後のマウス心臓の形態を比較したところ、心筋細胞面積の増加（心肥大）と間質の線維化がPPZ2投与によって有意に抑制されることがわかった。 Muscle LIM protein (MLP)-deficient mouse, a dilated cardiomyopathy model mouse (Knoll R, Hoshijima M, Hoffman HM, Person V, Lorenzen-Schmidt I, Bang ML, Hayashi T, Shiga N, Yasukawa H, Schaper W, McKenna W, Yokoyama M, Schork NJ, Omens JH, McCulloch AD, Kimura A, Gregorio CC, Poller W, Schaper J, Schultheiss HP and Chien KR The cardiac mechanical stretch sensor machinery involves a Z disc complex that is defective in a sub set of human dilated cardiomyopathy. Cell 111 (2002b), pp. 943-55.) develops heart failure 5 weeks after birth. As a result of measuring cardiac function (ejection fraction (EF) and left ventricular diameter fractional shortening (FS)) after implanting a PPZ2-containing osmotic pump in 8- to 9-week-old male mice, we found that the solvent Compared to MLP-deficient mice administered with only (50% (v/v) diethyl sulfoxide (DMSO)/ 50% (v/v) polyethylene glycol 300), cardiac function was significantly higher after 1 to 3 weeks. (Figure 2). Comparing the morphology of mouse hearts 3 weeks after PPZ2 administration, we found that PPZ2 administration significantly suppressed the increase in myocardial cell area (cardiac hypertrophy) and interstitial fibrosis.
試験例２
圧負荷誘発心不全モデルマウスにおけるTRPC6チャネル活性化
　圧負荷誘発心不全モデルマウス：129Sv系統（Kitajima N, et al. TRPC3 positively regulates reactive oxygen species driving maladaptive cardiac remodeling. Sci Rep 6, 37001 (2016)）の雄性マウス（10－16週令、WTとTRPC6-/-）の横行大動脈（transverse aortic constriction; TAC）を狭窄し、圧負荷を誘発した。TAC1週間後に心機能不全になったことを確認した後、PPZ2を含んだ浸透圧ポンプを腹腔内に埋込み、4週間の持続投与を行った。 Test example 2
TRPC6 channel activation in pressure overload-induced heart failure model mice Pressure overload-induced heart failure model mice: Male mice of the 129Sv strain (Kitajima N, et al. TRPC3 positively regulates reactive oxygen species driving maladaptive cardiac remodeling. Sci Rep 6, 37001 (2016)) The transverse aortic constriction (TAC) of (10-16 weeks old, WT and TRPC6-/-) was constricted and pressure overload was induced. After confirming cardiac dysfunction after 1 week of TAC, an osmotic pump containing PPZ2 was implanted intraperitoneally and continuous administration was performed for 4 weeks.
　得られた結果を図３に示す。TAC後の心不全モデルマウスの心臓では、TRPC6チャネル遺伝子の発現量が代償的に増加することが知られている（Kuwahara K, et al. TRPC6 fulfills a calcineurin signaling circuit during pathologic cardiac remodeling. Journal of Clinical Investigation 116, 3114-3126 (2006).）。 TAC１週間後、心機能が低下したことを確認してから、PPZ2を含んだ浸透圧ポンプを埋込み、その後の心機能変化を解析した。TAC後、対照マウスでは時間経過とともに心機能が徐々に低下したのに対し、PPZ投与マウスでは、投与後３～４週間のEF、FS上昇率が有意に増えていることがわかった（図3－１ａ,ｂ）。PPZ2投与4週間後のマウス心臓の形態を比較したところ、心筋細胞面積の増加（心肥大）と間質の線維化がPPZ2投与によって有意に抑制されることがわかった（図3－１ｃ,図３－２ｄ）。正常マウスにPPZ2を投与しても心筋組織内Zn2+濃度に変化はなかったが、TAC処置心筋において、PPZ2投与により有意なZn2+濃度増加が観察された（図３－２e）。この増加は、TRPC6欠損により完全に抑制された。また、β受容体が活性化すると内因性の心筋保護シグナルであるプロテインキナーゼG (PKG)が活性化されることも知られている（Takimoto E, et al. cGMP catabolism by phosphodiesterase 5A regulates cardiac adrenergic stimulation by NOS3-dependent mechanism. Circ Res 96, 100-109 (2005).）。PPZ2投与は、TAC処置心筋においてのみ有意な活性増加を引き起こし、この活性増加はTRPC6欠損によって抑制されることわかった（図３－２ｆ）。総じて、図３は、PPZ2投与マウスでは、溶媒（50% (v/v) ジエチルスルホキシド (DMSO)/ 50% (v/v) ポリエチレングリコール300）を投与した（対照）マウス（対照）と比べて、TAC後の心不全が有意に改善することを示している。 The results obtained are shown in FIG. It is known that the expression level of the TRPC6 channel gene increases compensatoryly in the hearts of heart failure model mice after TAC (Kuwahara K, et al. TRPC6 fulfills a calcineurin signaling circuit during pathologic cardiac remodeling. Journal of Clinical Investigation 116, 3114-3126 (2006). After confirming that cardiac function had decreased one week after TAC, an osmotic pump containing PPZ2 was implanted, and subsequent changes in cardiac function were analyzed. After TAC, cardiac function gradually decreased over time in control mice, whereas in PPZ-treated mice, the rate of increase in EF and FS increased significantly for 3 to 4 weeks after administration (Figure 3 -1a,b). When we compared the morphology of mouse hearts 4 weeks after PPZ2 administration, we found that the increase in myocardial cell area (cardiac hypertrophy) and interstitial fibrosis were significantly suppressed by PPZ2 administration (Figure 3-1c, 3-2d). Administration of PPZ2 to normal mice did not change the Zn 2+ concentration in myocardial tissue, but a significant increase in Zn 2+ concentration was observed in TAC-treated myocardium due to PPZ2 administration (Figure 3-2e). This increase was completely suppressed by TRPC6 deficiency. It is also known that protein kinase G (PKG), an endogenous cardioprotective signal, is activated when β receptors are activated (Takimoto E, et al. cGMP catabolism by phosphodiesterase 5A regulates cardiac adrenergic stimulation by NOS3-dependent mechanism. Circ Res 96, 100-109 (2005).) It was found that PPZ2 administration caused a significant activity increase only in TAC-treated myocardium, and this activity increase was suppressed by TRPC6 deficiency (Figure 3-2f). Overall, Fig. 3 shows that PPZ2-treated mice had higher levels of cancer compared to mice treated with vehicle (50% (v/v) diethyl sulfoxide (DMSO)/50% (v/v) polyethylene glycol 300) (control). , showing a significant improvement in heart failure after TAC.
試験例３
心筋梗塞後心不全モデルマウスにおけるTRPC6チャネル活性化
　心筋梗塞後心不全モデルマウス（Nishimura A, et al. Hypoxia-induced interaction of filamin with Drp1 causes mitochondrial hyperfission-associated myocardial senescence. Sci Signal 11, (2018).）において冠動脈左前下行枝を結紮し、心筋梗塞（myocardial infarction; MI）を誘発させた。MI１週間後、心機能が低下したことを確認してから、PPZ2を含んだ浸透圧ポンプを埋込み、その後の心機能変化を解析した。MI後、対照マウスでは時間経過とともに心機能が徐々に低下したのに対し、PPZ投与マウスでは、投与後３～４週間のEF、FS上昇率が有意に増えていることがわかった（図４）。 Test example 3
TRPC6 channel activation in post-myocardial infarction heart failure model mice (Nishimura A, et al. Hypoxia-induced interaction of filamin with Drp1 causes mitochondrial hyperfission-associated myocardial senescence. Sci Signal 11, (2018).) The left anterior descending coronary artery was ligated to induce myocardial infarction (MI). After confirming that cardiac function had decreased one week after MI, an osmotic pump containing PPZ2 was implanted, and subsequent changes in cardiac function were analyzed. After MI, cardiac function gradually decreased over time in control mice, whereas in PPZ-treated mice, the rate of increase in EF and FS increased significantly for 3 to 4 weeks after administration (Figure 4 ).
試験例４
一過性の血圧低下誘発による圧受容反射応答に対する効果
　PPZ2を30分前に腹腔内投与したマウスにヒドララジン(0.5 mg/kg, 1.2 mL/h)を投与し、一過性の血圧低下誘発による圧受容反射応答、特に心臓の陽性変力作用が増強するか検討した。TRPC6チャネル阻害剤BI749327 (30 mg/kg) （In vivo selective inhibition of TRPC6 by antagonist BI 749327 ameliorates fibrosis and dysfunction in cardiac and renal disease, https://doi.org/10.1073/pnas.1815354116）を、PPZ2 (2.5 mg/kg)投与する5時間前に経口投与した。 Test example 4
Effect on baroreceptive reflex response by inducing a transient decrease in blood pressure Hydralazine (0.5 mg/kg, 1.2 mL/h) was administered to mice that had been intraperitoneally administered PPZ2 30 minutes prior to inducing a transient decrease in blood pressure. We investigated whether baroreceptive reflex responses, especially cardiac positive inotropic effects, were enhanced. TRPC6 channel inhibitor BI749327 (30 mg/kg) (In vivo selective inhibition of TRPC6 by antagonist BI 749327 ameliorates fibrosis and dysfunction in cardiac and renal disease, https://doi.org/10.1073/pnas.1815354116) was combined with PPZ2 ( 2.5 mg/kg) was administered orally 5 hours before administration.
　得られた結果を図５に示す。縦軸はヒドララジン誘発性の（圧受容反射による）左心室収縮力の増加分（ΔLVdP/dtmax）または心拍数の増加分（ΔHeart Rate）を示している。PPZ2の投与により、PPZ2の濃度依存的にヒドララジン誘発性のΔLVdP/dtmaxが増加する。一方で、PPZ2投与ではヒドララジン誘発性のΔHeart Rateは変化しない。さらに、PPZ2の投与によるヒドララジン誘発性のΔLVdP/dtmaxの増加は、TRPC6チャネル阻害剤であるBI749327の投与により抑制された。以上のことから、PPZ2はTRPC6チャネルの活性化を介して濃度依存的に圧受容反射による心臓の陽性変力作用を増強することがわかった。 The obtained results are shown in Figure 5. The vertical axis shows the hydralazine-induced increase in left ventricular contractile force (ΔLVdP/dtmax) or heart rate (ΔHeart Rate). Administration of PPZ2 increases hydralazine-induced ΔLVdP/dtmax in a PPZ2 concentration-dependent manner. On the other hand, PPZ2 administration did not change the hydralazine-induced ΔHeart Rate. Furthermore, the hydralazine-induced increase in ΔLVdP/dtmax due to administration of PPZ2 was suppressed by administration of BI749327, a TRPC6 channel inhibitor. From the above, it was found that PPZ2 enhances the positive inotropic effect of the heart caused by the baroreceptive reflex in a concentration-dependent manner through activation of the TRPC6 channel.
試験例５
TRPC6チャネル活性化物質GSK1702934AのZn2+濃度増強効果
　PPZ2以外のTRPC6チャネル活性化物質GSK1702934A（In vivo selective inhibition of TRPC6 by antagonist BI 749327 ameliorates fibrosis and dysfunction in cardiac and renal disease, https://doi.org/10.1073/pnas.1815354116）によるZn2+濃度増強効果を調べた。HEK293細胞にTRPC6 (WT)またはZn2+透過能欠損変異体（KYD）を発現させ、TRPC6活性化物質処置により細胞内Zn2+濃度上昇がTRPC6依存的に誘発されるか検討した。 Test example 5
Zn 2+ concentration enhancement effect of TRPC6 channel activator GSK1702934A TRPC6 channel activator other than PPZ2 GSK1702934A (In vivo selective inhibition of TRPC6 by antagonist BI 749327 ameliorates fibrosis and dysfunction in cardiac and renal disease, https://doi.org /10.1073/pnas.1815354116) was investigated to enhance Zn 2+ concentration. We expressed TRPC6 (WT) or Zn 2+ permeability-defective mutant (KYD) in HEK293 cells, and examined whether treatment with a TRPC6 activator would induce an increase in intracellular Zn 2+ concentration in a TRPC6-dependent manner.
　得られた結果を図６に示す。細胞内Zn2+濃度をFluoZn-3染色にて検討した。TRPC6 (WT)を発現させた細胞ではPPZ2の処置により、FluoZn-3蛍光強度が増加した。一方で、TRPC6 (KYD)を発現させた細胞では、PPZ2の処置によりFluoZn-3蛍光強度は変化しなかった。さらに、GSK1702934A処置でもPPZ2同様に、TRPC6 (WT)を発現させた細胞ではFluoZn-3蛍光強度は増加したが、TRPC6 (KYD)を発現させた細胞ではFluoZn-3蛍光強度に変化はなかった。以上のことから、TRPC6チャネル活性化物質GSK1702934AはPPZ2同様に細胞内のZn2+濃度増強効果を有することが明らかになった。 The obtained results are shown in FIG. Intracellular Zn 2+ concentration was examined using FluoZn-3 staining. Treatment with PPZ2 increased FluoZn-3 fluorescence intensity in cells expressing TRPC6 (WT). On the other hand, in cells expressing TRPC6 (KYD), treatment with PPZ2 did not change FluoZn-3 fluorescence intensity. Furthermore, with GSK1702934A treatment, as with PPZ2, FluoZn-3 fluorescence intensity increased in cells expressing TRPC6 (WT), but there was no change in FluoZn-3 fluorescence intensity in cells expressing TRPC6 (KYD). From the above, it was revealed that the TRPC6 channel activator GSK1702934A has the effect of enhancing intracellular Zn 2+ concentration similarly to PPZ2.
試験例６
TRPC6チャネル活性化物質GSK1702934Aおよびハイパーフォリンの心筋収縮力増強効果
　PPZ2以外のTRPC6チャネル活性化物質GSK1702934Aおよびハイパーフォリン（hyperforin）（GSK1702934A and M085 directly activate TRPC6 via a mechanism of stimulating the extracellular cavity formed by the pore helix and transmembrane helix S6, https://doi.org/10.1016/j.jbc.2021.101125）による心筋収縮力増強効果を調べた。新生仔ラット心筋細胞にTRPC6活性化物質を30分間処置し、ノルエピネフリ（NE）（1μM）による心筋細胞の収縮力の上昇が認められるか検討した。 Test example 6
Effect of TRPC6 channel activator GSK1702934A and hyperforin on enhancing myocardial contractility TRPC6 channel activator other than PPZ2 GSK1702934A and hyperforin (GSK1702934A and M085 directly activate TRPC6 via a mechanism of stimulating the extracellular cavity formed by the pore helix and transmembrane helix S6, https://doi.org/10.1016/j.jbc.2021.101125) to investigate the effect of enhancing myocardial contractility. Neonatal rat cardiomyocytes were treated with a TRPC6 activator for 30 minutes to examine whether norepinephrine (NE) (1 μM) increased the contractile force of cardiomyocytes.
　得られた結果を図７に示す。PPZ2はNEによる心筋収縮を増強した。また、PPZ2による心筋収縮増強作用はZn2+のキレーターであるTPEN 処置により減弱した。したがって、PPZ2はZn2+依存的にNEによる心筋収縮を増強することが明らかとなった（図７a）。GSK1702934AとハイパーフォリンもPPZ2同様の効果を示すことがわかった（図７b）。 The obtained results are shown in FIG. PPZ2 enhanced NE-induced myocardial contraction. In addition, the myocardial contraction enhancing effect of PPZ2 was attenuated by treatment with TPEN, a Zn 2+ chelator. Therefore, it was revealed that PPZ2 enhances NE-induced myocardial contraction in a Zn 2+ -dependent manner (Fig. 7a). It was found that GSK1702934A and hyperforin also showed effects similar to PPZ2 (Figure 7b).
試験例７
筋ジストロフィーモデルマウスにおけるTRPC6チャネル活性化
　筋ジストロフィーモデルマウス：C57BL/10-mdxマウス（6週齢：日本クレアから購入）にPPZ2(2.5 mg/kg)を含んだ浸透圧ポンプを腹腔内に埋込み、4週間の持続投与を行った。PPZ2投与マウスでは、溶媒（50% (v/v) ジエチルスルホキシド (DMSO)/ 50% (v/v) ポリエチレングリコール300）を投与したマウス（対照）と比べて、筋障害マーカーである血中のLDH(乳酸デヒドロゲナーゼ)やCK(クレアチニンキナーゼ)が有意に抑制されることがわかった（図８）。 Test example 7
TRPC6 channel activation in muscular dystrophy model mice Muscular dystrophy model mice: C57BL/10-mdx mice (6 weeks old, purchased from CLEA Japan) were implanted with an osmotic pump containing PPZ2 (2.5 mg/kg) intraperitoneally for 4 weeks. Continuous administration was performed. In mice treated with PPZ2, blood levels, which are markers of myopathy, were significantly reduced compared to mice treated with vehicle (50% (v/v) diethyl sulfoxide (DMSO)/50% (v/v) polyethylene glycol 300) (control). It was found that LDH (lactate dehydrogenase) and CK (creatinine kinase) were significantly inhibited (Figure 8).
　以下、調製例および実施例を参照して本発明化合物の合成例を具体的に説明するが、本発明はこれらに限定されない。なお、化合物のＬＣ／ＭＳ分析には以下の条件を用いた。
［ＬＣ／ＭＳ分析条件］
ＬＣ／ＭＳシステム：Waters ACQUITY UPLC H-Class/QDa　Sample Manager - FTN
　Quaternary Solvent Manager
　Column Heater A
　PDAeλ Detector
　QDa Detector
カラム：ACQUITY UPLC BEH C18 1.7 μm (2.1×50mm)流速：0.5 mL/min
溶出条件：移動相Ａ；アセトニトリル、移動相Ｂ；０．１％ぎ酸水溶液 Synthesis examples of the compounds of the present invention will be specifically explained below with reference to Preparation Examples and Examples, but the present invention is not limited thereto. Note that the following conditions were used for LC/MS analysis of the compound.
[LC/MS analysis conditions]
LC/MS System: Waters ACQUITY UPLC H-Class/QDa Sample Manager - FTN
Quaternary Solvent Manager
Column Heater A
PDAeλ Detector
QDa Detector
Column: ACQUITY UPLC BEH C18 1.7 μm (2.1×50mm) Flow rate: 0.5 mL/min
Elution conditions: Mobile phase A: acetonitrile, mobile phase B: 0.1% formic acid aqueous solution
表１
Figure JPOXMLDOC01-appb-T000019
 Table 1
Figure JPOXMLDOC01-appb-T000019
調製例１
２－ブロモ－Ｎ－（２－エトキシフェニル）アセトアミド
Figure JPOXMLDOC01-appb-C000020
 　２－エトキシアニリン（1.37 g、10 mmol）とトリエチルアミン（1.33 mL、10 mmol）のジクロロメタン（10 mL）の溶液に、ブロモ酢酸ブロミド（0.87 mL、10 mmol）を０℃で滴下した。反応液を窒素雰囲気下、室温で３時間撹拌し、飽和食塩水を加え、酢酸エチルで抽出した。有機層を飽和炭酸水素ナトリウム水溶液で洗浄し、硫酸ナトリウムで乾燥し、濃縮して目的化合物（2.587 g）を得た。
１H NMR (CDCl３, 400 MHz) δ8.96 (br, 1 H), 8.31 (dd, 1 H, J = 2.0, 8.0 Hz), 7.07 (dt, 1 H, J = 2.0, 8.0 Hz), 6.97 (dt, 1 H, J = 1.2, 8.0 Hz), 6.87 (dd, 1 H, J = 1.2, 8.0 Hz), 4.12 (q, 2 H, J = 6.8 Hz), 4.05 (s, 2 H), 1.48 (t, 3 H, J = 6.8 Hz), 
LC/MS: r.t. 2.58 min, m/z 258.0, 260.0 (M+ + 1). Preparation example 1
2-Bromo-N-(2-ethoxyphenyl)acetamide
Figure JPOXMLDOC01-appb-C000020
 Bromoacetic acid bromide (0.87 mL, 10 mmol) was added dropwise to a solution of 2-ethoxyaniline (1.37 g, 10 mmol) and triethylamine (1.33 mL, 10 mmol) in dichloromethane (10 mL) at 0°C. The reaction solution was stirred at room temperature under nitrogen atmosphere for 3 hours, saturated brine was added, and the mixture was extracted with ethyl acetate. The organic layer was washed with saturated aqueous sodium bicarbonate solution, dried over sodium sulfate, and concentrated to obtain the target compound (2.587 g).
1 H NMR (CDCl 3 , 400 MHz) δ8.96 (br, 1 H), 8.31 (dd, 1 H, J = 2.0, 8.0 Hz), 7.07 (dt, 1 H, J = 2.0, 8.0 Hz), 6.97 (dt, 1 H, J = 1.2, 8.0 Hz), 6.87 (dd, 1 H, J = 1.2, 8.0 Hz), 4.12 (q, 2 H, J = 6.8 Hz), 4.05 (s, 2 H) , 1.48 (t, 3 H, J = 6.8 Hz),
LC/MS: rt 2.58 min, m/z 258.0, 260.0 (M + + 1).
調製例２
２－ブロモ－Ｎ－（２－（トリフルオロメトキシ）フェニル）アセトアミド
Figure JPOXMLDOC01-appb-C000021
 　２－トリフルオロメトキシアニリン（354 mg、2.2 mmol）とトリエチルアミン（306 μL、2.2 mmol）のジクロロメタン（5 mL）の溶液に、ブロモ酢酸ブロミド（174 μL、2 mmol）を０℃で滴下した。反応液を窒素雰囲気下室温で３時間撹拌し、飽和塩化アンモニウム水溶液を加え、酢酸エチルで抽出した。有機層を飽和炭酸水素ナトリウム水溶液で洗浄し、硫酸ナトリウムで乾燥し、濃縮して目的化合物（607 mg）を得た。
１H NMR (CDCl３, 400 MHz) δ8.63 (brs, 1 H), 8.36 (dd, 1 H, J = 1.2, 8.0 Hz), 7.32 (dt, 1 H, J = 1.2, 8.0 Hz), 7.29 (dt, 1 H, J = 8.0, 1.2 Hz), 7.17 (dt, 1 H, J = 1.6, 8.0 Hz), 4.12 (q, 2 H, J = 6.8 Hz), 4.06 (s, 2 H), 
LC/MS: r.t. 2.61 min, m/z 297.9, 300 (M+ + 1). Preparation example 2
2-Bromo-N-(2-(trifluoromethoxy)phenyl)acetamide
Figure JPOXMLDOC01-appb-C000021
 Bromoacetic acid bromide (174 μL, 2 mmol) was added dropwise to a solution of 2-trifluoromethoxyaniline (354 mg, 2.2 mmol) and triethylamine (306 μL, 2.2 mmol) in dichloromethane (5 mL) at 0°C. The reaction solution was stirred at room temperature under a nitrogen atmosphere for 3 hours, a saturated aqueous ammonium chloride solution was added, and the mixture was extracted with ethyl acetate. The organic layer was washed with saturated aqueous sodium bicarbonate solution, dried over sodium sulfate, and concentrated to obtain the target compound (607 mg).
1 H NMR (CDCl 3 , 400 MHz) δ8.63 (brs, 1 H), 8.36 (dd, 1 H, J = 1.2, 8.0 Hz), 7.32 (dt, 1 H, J = 1.2, 8.0 Hz), 7.29 (dt, 1 H, J = 8.0, 1.2 Hz), 7.17 (dt, 1 H, J = 1.6, 8.0 Hz), 4.12 (q, 2 H, J = 6.8 Hz), 4.06 (s, 2 H) ,
LC/MS: rt 2.61 min, m/z 297.9, 300 (M + + 1).
調製例３
２－ブロモ－Ｎ－（２－（２，２，２－トリフルオロエトキシ）フェニル）アセトアミド
Figure JPOXMLDOC01-appb-C000022
 　２－（２，２，２－トリフルオロエトキシ）アニリン（379 mg、2 mmol）とトリエチルアミン（306 μL、2.2 mmol）のジクロロメタン（5 mL）の溶液に、ブロモ酢酸ブロミド（174 μL、2 mmol）を０℃で滴下した。反応液を窒素雰囲気下室温で３時間撹拌し、飽和炭酸水素ナトリウム水溶液を加え、酢酸エチルで抽出した。有機層を飽和炭酸水素ナトリウム水溶液で洗浄し、硫酸ナトリウムで乾燥し、濃縮して目的化合物（550.8 mg）を得た。
１H NMR (CDCl３, 400 MHz) δ8.87 (brs, 1 H), 8.32 - 8.39 (m, 1 H), 7.05 - 7.15 (m, 2 H), 6.86 - 6.91 (m, 1 H), 4.44 (q, 2 H, J = 7.6 Hz), 4.04 (s, 2 H), 
LC/MS: r.t. 2.60 min, m/z 312.0, 314 (M+ + 1). Preparation example 3
2-Bromo-N-(2-(2,2,2-trifluoroethoxy)phenyl)acetamide
Figure JPOXMLDOC01-appb-C000022
 Bromoacetic acid bromide (174 μL, 2 mmol) was added to a solution of 2-(2,2,2-trifluoroethoxy)aniline (379 mg, 2 mmol) and triethylamine (306 μL, 2.2 mmol) in dichloromethane (5 mL). was added dropwise at 0°C. The reaction solution was stirred at room temperature under nitrogen atmosphere for 3 hours, saturated aqueous sodium hydrogen carbonate solution was added, and the mixture was extracted with ethyl acetate. The organic layer was washed with saturated aqueous sodium bicarbonate solution, dried over sodium sulfate, and concentrated to obtain the target compound (550.8 mg).
1 H NMR (CDCl 3 , 400 MHz) δ8.87 (brs, 1 H), 8.32 - 8.39 (m, 1 H), 7.05 - 7.15 (m, 2 H), 6.86 - 6.91 (m, 1 H), 4.44 (q, 2 H, J = 7.6 Hz), 4.04 (s, 2 H),
LC/MS: rt 2.60 min, m/z 312.0, 314 (M + + 1).
調製例４
４－（２－クロロ－4－（メトキシカルボニル）フェニル）ピペラジン－1－カルボン酸tert－ブチル
Figure JPOXMLDOC01-appb-C000023
 　ピペラジン－１－カルボン酸tert－ブチル（4..91 g, 26.36 mmol）、３－クロロー４－フルオロ安息香酸メチル（4.836 g, 25.64 mmol）、およびDBU（4590 μL, 30.75 mmol）のDMSO（30 mL）の溶液を窒素雰囲気下100℃で10時間加熱した。反応液を室温まで冷却し飽和塩化アンモニウム水溶液を加え、酢酸エチルで抽出した。有機層を飽和食塩水で洗浄、硫酸ナトリウムで乾燥し、濃縮した。残渣を中圧分取シリカゲルカラムクロマトグラフィー（ヘキサン/酢酸エチル）で精製し、目的物（5.12 g）の結晶を得た。収率56%。
Rf = 0.38 (5 : 1 hexane/ethyl acetate), １H NMR (CDCl３, 400 MHz) δ8.02 (d, 1 H, J = 2 Hz), 7.87 (dd, 1 H, J = 2.0, 8.0 Hz), 6.99 (d, 1 H, J = 8.0 Hz), 3.88 (s, 3 H),  3.54 - 3.64 (br, 4 H), 3.02 - 3.12 (br, 4H), 1.48 (s, 9 H), 
LC/MS: r.t. = 3.34 min, m/z 355.1 (M+ + 1). Preparation example 4
tert-butyl 4-(2-chloro-4-(methoxycarbonyl)phenyl)piperazine-1-carboxylate
Figure JPOXMLDOC01-appb-C000023
 tert-butyl piperazine-1-carboxylate (4..91 g, 26.36 mmol), methyl 3-chloro-4-fluorobenzoate (4.836 g, 25.64 mmol), and DBU (4590 μL, 30.75 mmol) in DMSO (30 mL) solution was heated at 100 °C for 10 h under nitrogen atmosphere. The reaction solution was cooled to room temperature, a saturated aqueous ammonium chloride solution was added, and the mixture was extracted with ethyl acetate. The organic layer was washed with saturated brine, dried over sodium sulfate, and concentrated. The residue was purified by medium pressure preparative silica gel column chromatography (hexane/ethyl acetate) to obtain crystals of the desired product (5.12 g). Yield 56%.
Rf = 0.38 (5 : 1 hexane/ethyl acetate), 1 H NMR (CDCl 3 , 400 MHz) δ8.02 (d, 1 H, J = 2 Hz), 7.87 (dd, 1 H, J = 2.0, 8.0 Hz), 6.99 (d, 1 H, J = 8.0 Hz), 3.88 (s, 3 H), 3.54 - 3.64 (br, 4 H), 3.02 - 3.12 (br, 4H), 1.48 (s, 9 H) ,
LC/MS: rt = 3.34 min, m/z 355.1 (M + + 1).
調製例５
4－（4－（tert－ブトキシカルボニル）ピペラジン－１－イル）―３－クロロ安息香酸
Figure JPOXMLDOC01-appb-C000024
 　4－（4－（tert－ブトキシカルボニル）ピペラジン－１－イル）―３－クロロ安息香酸メチルエステル（3.31 g, 9.33 mmol）の1，4－ジオキサン (20 mL)の溶液に1.1 N水酸化ナトリウム水溶液（20 mL, 22 mmol）を加え、窒素雰囲気下、室温で終夜撹拌した。反応液を1 N塩酸でpH約４まで中和し、酢酸エチルで抽出した。有機層を硫酸ナトリウムで乾燥し、濃縮乾固し、目的物（3.68 g）を得た。収率100％。
１H NMR (CDCl３, 400 MHz) δ8.10 (d, 1 H, J = 2.0 Hz), 7.95 (dd, 1 H, J = 2.0, 8.8 Hz), 7.03 (d, 1 H, J = 8.8 Hz), 3.54 - 3.69 (br, 4 H), 3.02 - 3.19 (br, 4H), 1.49 (s, 9 H),
LC/MS R.T. = 2.84 min, m/z 341.1 (M+ + 1). Preparation example 5
4-(4-(tert-butoxycarbonyl)piperazin-1-yl)-3-chlorobenzoic acid
Figure JPOXMLDOC01-appb-C000024
 A solution of 4-(4-(tert-butoxycarbonyl)piperazin-1-yl)-3-chlorobenzoic acid methyl ester (3.31 g, 9.33 mmol) in 1,4-dioxane (20 mL) in 1.1 N sodium hydroxide An aqueous solution (20 mL, 22 mmol) was added, and the mixture was stirred at room temperature overnight under a nitrogen atmosphere. The reaction solution was neutralized to pH approximately 4 with 1 N hydrochloric acid, and extracted with ethyl acetate. The organic layer was dried over sodium sulfate and concentrated to dryness to obtain the desired product (3.68 g). Yield 100%.
1 H NMR (CDCl 3 , 400 MHz) δ8.10 (d, 1 H, J = 2.0 Hz), 7.95 (dd, 1 H, J = 2.0, 8.8 Hz), 7.03 (d, 1 H, J = 8.8 Hz), 3.54 - 3.69 (br, 4 H), 3.02 - 3.19 (br, 4H), 1.49 (s, 9 H),
LC/MS RT = 2.84 min, m/z 341.1 (M + + 1).
調製例６
４－（２－クロロ－４－（４，４－ジフルオロピぺリジン－１－カルボニル）フェニル）ピペラジン－１－カルボン酸tert-ブチルエステル
Figure JPOXMLDOC01-appb-C000025
 　４，４－ジフルオロピペリジン－２塩酸塩（105.5 mg, 0.67 mmol）、調製例５にて調製した4－（4－（tert－ブトキシカルボニル）ピペラジン－１－イル）―３－クロロ安息香酸（228.6 mg, 0.67 mmol）、HATU (267.7 mg, 0.704 mmol)とジイソプロピルエチルアミン（280 μmol, 1.61 mmol）のDMF (3 mL)の溶液を窒素雰囲気下室温で3時間撹拌した。反応液に飽和塩化アンモニウム水溶液を加え、酢酸エチルで抽出した。有機層を硫酸ナトリウムで乾燥し、濃縮した。残渣を中圧分取シリカゲルカラムクロマトグラフィー（ヘキサン・酢酸エチル）で精製し、目的物（272.4 mg）の結晶を得た。収率92%。
Rf = 0.59 (2 : 1 = ヘキサン/酢酸エチル), 
１H NMR (CDCl３, 400 MHz) δ7.46 (d, 1 H, J = 1.6 Hz), 7.29 (dd, 1 H, J = 2.0, 8.4 Hz), 7.02 (d, 1 H, J = 8.0 Hz), 3.6 - 3.9 (br, 4 H), 3.55 - 3.65 (br. 4 H), 2.96 - 3.07 (br, 4H), 1.90 - 2.10 (br, 4 H) 1.49 (s, 9 H), 
LC/MS: r.t. 3.11 min, m/z 444.2 (M+ + 1). Preparation example 6
4-(2-chloro-4-(4,4-difluoropiperidine-1-carbonyl)phenyl)piperazine-1-carboxylic acid tert-butyl ester
Figure JPOXMLDOC01-appb-C000025
 4,4-difluoropiperidine-dihydrochloride (105.5 mg, 0.67 mmol), 4-(4-(tert-butoxycarbonyl)piperazin-1-yl)-3-chlorobenzoic acid (228.6 A solution of HATU (267.7 mg, 0.704 mmol) and diisopropylethylamine (280 μmol, 1.61 mmol) in DMF (3 mL) was stirred at room temperature under nitrogen atmosphere for 3 hours. A saturated aqueous ammonium chloride solution was added to the reaction mixture, and the mixture was extracted with ethyl acetate. The organic layer was dried over sodium sulfate and concentrated. The residue was purified by medium pressure preparative silica gel column chromatography (hexane/ethyl acetate) to obtain crystals of the desired product (272.4 mg). Yield 92%.
Rf = 0.59 (2 : 1 = hexane/ethyl acetate),
1 H NMR (CDCl 3 , 400 MHz) δ7.46 (d, 1 H, J = 1.6 Hz), 7.29 (dd, 1 H, J = 2.0, 8.4 Hz), 7.02 (d, 1 H, J = 8.0 Hz), 3.6 - 3.9 (br, 4 H), 3.55 - 3.65 (br. 4 H), 2.96 - 3.07 (br, 4H), 1.90 - 2.10 (br, 4 H) 1.49 (s, 9 H),
LC/MS: rt 3.11 min, m/z 444.2 (M + + 1).
調製例７
（３－クロロ－4－（ピペラジン－1－イル）フェニル）（４，４－ジフルオロピペリジン－１－イル）メタノン二塩酸塩
Figure JPOXMLDOC01-appb-C000026
 　調製例６にて調製した４－（２－クロロ－４－（４，４－ジフルオロピぺリジン－１－カルボニル）フェニル）ピペラジン－１－カルボン酸tert-ブチルエステル (1933 mg, 4.35 mmol)の1,4-ジオキサン (5 mL)の溶液に4 N HCl／1,4-ジオキサン溶液（10 mL）を加え、室温で一晩撹拌した。混合物をエバポレーターで濃縮し、残渣に酢酸エチルを加え、ろ過した。得られた固体を酢酸エチルで洗浄し、減圧乾燥し、目的物（1564 mg）を得た。収率86％。
１H NMR (DMSO, 400 MHz) δ9.16 (br, 2 H), 7.56 (d, 1 H, J = 2.0 Hz), 7.42 (dd, 1 H, J = 2.0, 8.0 Hz), 7.24 (d, 1 H, J = 8.0 Hz), 3.40 - 3.70 (br, 4 H), 3.13 - 3.28 (br. 8 H), 2.69 (s, 1 H), 1.91 - 2.11 (br, 4 H), 
LC/MS: r.t. 1.59 min, m/z 344.1 (M+ + 1). Preparation example 7
(3-chloro-4-(piperazin-1-yl)phenyl)(4,4-difluoropiperidin-1-yl)methanone dihydrochloride
Figure JPOXMLDOC01-appb-C000026
 1 of 4-(2-chloro-4-(4,4-difluoropiperidine-1-carbonyl)phenyl)piperazine-1-carboxylic acid tert-butyl ester (1933 mg, 4.35 mmol) prepared in Preparation Example 6 A 4 N HCl/1,4-dioxane solution (10 mL) was added to a solution of ,4-dioxane (5 mL), and the mixture was stirred at room temperature overnight. The mixture was concentrated using an evaporator, ethyl acetate was added to the residue, and the mixture was filtered. The obtained solid was washed with ethyl acetate and dried under reduced pressure to obtain the desired product (1564 mg). Yield 86%.
1 H NMR (DMSO, 400 MHz) δ9.16 (br, 2 H), 7.56 (d, 1 H, J = 2.0 Hz), 7.42 (dd, 1 H, J = 2.0, 8.0 Hz), 7.24 (d , 1 H, J = 8.0 Hz), 3.40 - 3.70 (br, 4 H), 3.13 - 3.28 (br. 8 H), 2.69 (s, 1 H), 1.91 - 2.11 (br, 4 H),
LC/MS: rt 1.59 min, m/z 344.1 (M + + 1).
実施例１
２－（４－（2－クロロ－4－（４，４－ジフルオロピぺリジン－1－カルボニル）フェニル）ピペラジン－1－イル）―N－（2－エトキシフェニル）アセトアミド
Figure JPOXMLDOC01-appb-C000027
 　調製例７にて調製した（３－クロロ－4－（ピペラジン－1－イル）フェニル）（４，４－ジフルオロピペリジン－１－イル）メタノン二塩酸塩(55 mg, 0.13 mmol)、調製例１にて調製した２－ブロモ－Ｎ－（２－エトキシフェニル）アセトアミド (52 mg, 0.2 mmol)、およびDBU (60 μL, 0.4 mmol)のDMF（1 mL）の溶液を窒素雰囲気下、80℃で10時間加熱撹拌した。混合物を室温に戻し、飽和塩化アンモニウム水溶液を加え、酢酸エチルで抽出し、有機層を飽和炭酸水素ナトリウム水溶液で洗浄し、硫酸ナトリウムで乾燥し、濃縮した。残渣を中圧分取シリカゲルカラムクロマトグラフィー（ヘキサン/酢酸エチル）で精製し、目的物（57.3 mg）の結晶を得た。収率85%。
Rf = 0.34 (1 : 1 ヘキサン/酢酸エチル), 
１H NMR (CDCl３, 400 MHz) δ9.86 (bs, 1 H), 8.44 (dd, 1 H, J = 1.6, 8.0 Hz), 7.46 (d, 1 H, J = 1.6 Hz), 7.32 (dd, 1 H, J = 2.0, 8.4 Hz), 7.05 (d, 1 H, J = 8.4 Hz), 7.03 (dd, 1 H, J = 2.0, 8.4 Hz), 6.97 (dt, 1 H, J = 1.6, 8.0 Hz), 6.87 (dd, 1 H, J = 1.6, 8.0 Hz), 4.11 (q, 2 H, J = 6.8 Hz), 3.50 - 3.92 (br, 4 H), 3.26 (s, 2 H), 3.12 - 3.26 (br, 4H), 2.81 - 2.90 (brt, 4 H), 1.90 - 2.12 (br, 4 H) 1.51 (t, 3 H, J = 6.8 Hz), 
LC/MS: r.t. 2.34 min, m/z 521.1 (M+ + 1). Example 1
2-(4-(2-chloro-4-(4,4-difluoropiperidine-1-carbonyl)phenyl)piperazin-1-yl)-N-(2-ethoxyphenyl)acetamide
Figure JPOXMLDOC01-appb-C000027
 (3-chloro-4-(piperazin-1-yl)phenyl)(4,4-difluoropiperidin-1-yl)methanone dihydrochloride (55 mg, 0.13 mmol) prepared in Preparation Example 7, Preparation Example 1 A solution of 2-bromo-N-(2-ethoxyphenyl)acetamide (52 mg, 0.2 mmol) prepared in The mixture was heated and stirred for 10 hours. The mixture was returned to room temperature, saturated ammonium chloride aqueous solution was added, and extracted with ethyl acetate. The organic layer was washed with saturated sodium bicarbonate aqueous solution, dried over sodium sulfate, and concentrated. The residue was purified by medium pressure preparative silica gel column chromatography (hexane/ethyl acetate) to obtain crystals of the desired product (57.3 mg). Yield 85%.
Rf = 0.34 (1 : 1 hexane/ethyl acetate),
1 H NMR (CDCl 3 , 400 MHz) δ9.86 (bs, 1 H), 8.44 (dd, 1 H, J = 1.6, 8.0 Hz), 7.46 (d, 1 H, J = 1.6 Hz), 7.32 ( dd, 1 H, J = 2.0, 8.4 Hz), 7.05 (d, 1 H, J = 8.4 Hz), 7.03 (dd, 1 H, J = 2.0, 8.4 Hz), 6.97 (dt, 1 H, J = 1.6, 8.0 Hz), 6.87 (dd, 1 H, J = 1.6, 8.0 Hz), 4.11 (q, 2 H, J = 6.8 Hz), 3.50 - 3.92 (br, 4 H), 3.26 (s, 2 H ), 3.12 - 3.26 (br, 4H), 2.81 - 2.90 (brt, 4 H), 1.90 - 2.12 (br, 4 H) 1.51 (t, 3 H, J = 6.8 Hz),
LC/MS: rt 2.34 min, m/z 521.1 (M + + 1).
実施例２
2－（4－（2－クロロ－４－（４，４－ジフルオロピぺリジン－1－カルボニル）フェニル）ピペラジン－1－イル）－N－（２－（トリフルオロメトキシ）フェニル）アセトアミド
Figure JPOXMLDOC01-appb-C000028
 　調製例７にて調製した（３－クロロ－4－（ピペラジン－1－イル）フェニル）（４，４－ジフルオロピペリジン－１－イル）メタノン二塩酸塩(41.7 mg, 0.1 mmol)、調製例２にて調製した２－ブロモ－Ｎ－（２－（トリフルオロメトキシ）フェニル）アセトアミド (29.8 mg, 0.1 mmol)、およびDBU (60 μL, 0.4 mmol)のDMF（1 mL）の溶液を窒素雰囲気下、80℃で10時間加熱撹拌した。混合物を室温に戻し、飽和塩化アンモニウム水溶液を加え、酢酸エチルで抽出し、有機層を飽和炭酸水素ナトリウム水溶液、続いて飽和食塩水で洗浄し、硫酸ナトリウムで乾燥し、濃縮した。残渣を中圧分取シリカゲルカラムクロマトグラフィー（ヘキサン/酢酸エチル）で精製し、目的物（25.3 mg）の結晶を得た。収率45%。
Rf = 0.22 (2 : 1 ヘキサン/酢酸エチル), 
１H NMR (CDCl３, 400 MHz) δ9.85 (bs, 1 H), 8.53 (dd, 1 H, J = 1.6, 8.0 Hz), 7.46 (d, 1 H, J = 2.0 Hz), 7.26 - 7.34 (m, 3 H), 7.11 (dt, 1 H, J = 1.2, 8.0 Hz), 7.07 (d, 1 H, J = 8.0 Hz), 3.50 - 3.92 (br, 4 H), 3.27 (s, 2 H), 3.05 - 3.25 (br, 4H), 2.80 - 2.88 (brt, 4 H), 1.90 - 2.12 (br, 4 H), 
LC/MS: r.t. 2.56 min, m/z 561.1 (M+ + 1). Example 2
2-(4-(2-chloro-4-(4,4-difluoropiperidine-1-carbonyl)phenyl)piperazin-1-yl)-N-(2-(trifluoromethoxy)phenyl)acetamide
Figure JPOXMLDOC01-appb-C000028
 (3-chloro-4-(piperazin-1-yl)phenyl)(4,4-difluoropiperidin-1-yl)methanone dihydrochloride (41.7 mg, 0.1 mmol) prepared in Preparation Example 7, Preparation Example 2 A solution of 2-bromo-N-(2-(trifluoromethoxy)phenyl)acetamide (29.8 mg, 0.1 mmol) and DBU (60 μL, 0.4 mmol) in DMF (1 mL) was added under a nitrogen atmosphere. The mixture was heated and stirred at 80°C for 10 hours. The mixture was cooled to room temperature, saturated ammonium chloride aqueous solution was added, and extracted with ethyl acetate. The organic layer was washed with saturated sodium bicarbonate aqueous solution and then with saturated brine, dried over sodium sulfate, and concentrated. The residue was purified by medium pressure preparative silica gel column chromatography (hexane/ethyl acetate) to obtain crystals of the desired product (25.3 mg). Yield 45%.
Rf = 0.22 (2 : 1 hexane/ethyl acetate),
1 H NMR (CDCl 3 , 400 MHz) δ9.85 (bs, 1 H), 8.53 (dd, 1 H, J = 1.6, 8.0 Hz), 7.46 (d, 1 H, J = 2.0 Hz), 7.26 - 7.34 (m, 3 H), 7.11 (dt, 1 H, J = 1.2, 8.0 Hz), 7.07 (d, 1 H, J = 8.0 Hz), 3.50 - 3.92 (br, 4 H), 3.27 (s, 2H), 3.05 - 3.25 (br, 4H), 2.80 - 2.88 (brt, 4H), 1.90 - 2.12 (br, 4H),
LC/MS: rt 2.56 min, m/z 561.1 (M + + 1).
実施例３
２－（４－（2－クロロ－4－（４，４－ジフルオロピぺリジン－1－カルボニル）フェニル）ピペラジン－1－イル）－N－（2－（2，2，2－トリフルオロエトキシ）フェニル）アセトアミド
Figure JPOXMLDOC01-appb-C000029
 　調製例７にて調製した（３－クロロ－4－（ピペラジン－1－イル）フェニル）（４，４－ジフルオロピペリジン－１－イル）メタノン二塩酸塩(41.7 mg, 0.1 mmol)、調製例３にて調製した２－ブロモ－Ｎ－（２－（２,２，２－トリフルオロエトキシ）フェニル）アセトアミド(31.2 mg, 0.1 mmol)、DBU (54 μL, 0.4 mmol)のDMF（1 mL）の溶液を窒素雰囲気下、80℃で10時間加熱撹拌した。反応液を室温に戻し、飽和塩化アンモニウム水溶液を加え、酢酸エチルで抽出し、有機層を飽和炭酸水素ナトリウム水溶液、つづいて飽和食塩水で洗浄し、硫酸ナトリウムで乾燥し、濃縮した。残渣を中圧分取シリカゲルカラムクロマトグラフィー（ヘキサン/酢酸エチル）で精製し、目的物（14.8 mg）の結晶を得た。収率26%。
Rf = 0.41 (1 : 1 ヘキサン/酢酸エチル), 
１H NMR (CDCl３, 400 MHz) δ9.83 (bs, 1 H), 8.51 - 8.55 (m, 1 H), 7.46 (d, 1 H, J = 1.6 Hz), 7.32 (dd, 1 H, J = 2.0, 8.4 Hz), 7.09 (dt, 1 H, J = 1.6, 7.6 Hz), 7.07 (d, 1 H, J = 6.8 Hz), 7.06 (d, 1 H, J = 7.2 Hz), 6.82 - 6.86 (m, 1 H), 4.43 (q, 2 H, J = 7.6 Hz), 3.50 - 3.94 (br, 4 H), 3.26 (s, 2 H), 3.10 - 3.23 (br, 4H), 2.80 - 2.88 (brt, 4 H), 1.90 - 2.12 (br, 4 H), 
LC/MS: r.t. 2.44 min, m/z 575.1 (M+ + 1). Example 3
2-(4-(2-chloro-4-(4,4-difluoropiperidine-1-carbonyl)phenyl)piperazin-1-yl)-N-(2-(2,2,2-trifluoroethoxy) phenyl)acetamide
Figure JPOXMLDOC01-appb-C000029
 (3-chloro-4-(piperazin-1-yl)phenyl)(4,4-difluoropiperidin-1-yl)methanone dihydrochloride (41.7 mg, 0.1 mmol) prepared in Preparation Example 7, Preparation Example 3 2-bromo-N-(2-(2,2,2-trifluoroethoxy)phenyl)acetamide (31.2 mg, 0.1 mmol) and DBU (54 μL, 0.4 mmol) in DMF (1 mL). The solution was heated and stirred at 80° C. for 10 hours under a nitrogen atmosphere. The reaction solution was returned to room temperature, a saturated aqueous ammonium chloride solution was added, and extraction was performed with ethyl acetate. The organic layer was washed with a saturated aqueous sodium bicarbonate solution and then with saturated brine, dried over sodium sulfate, and concentrated. The residue was purified by medium pressure preparative silica gel column chromatography (hexane/ethyl acetate) to obtain crystals of the desired product (14.8 mg). Yield 26%.
Rf = 0.41 (1 : 1 hexane/ethyl acetate),
1 H NMR (CDCl 3 , 400 MHz) δ9.83 (bs, 1 H), 8.51 - 8.55 (m, 1 H), 7.46 (d, 1 H, J = 1.6 Hz), 7.32 (dd, 1 H, J = 2.0, 8.4 Hz), 7.09 (dt, 1 H, J = 1.6, 7.6 Hz), 7.07 (d, 1 H, J = 6.8 Hz), 7.06 (d, 1 H, J = 7.2 Hz), 6.82 - 6.86 (m, 1 H), 4.43 (q, 2 H, J = 7.6 Hz), 3.50 - 3.94 (br, 4 H), 3.26 (s, 2 H), 3.10 - 3.23 (br, 4H), 2.80 - 2.88 (brt, 4H), 1.90 - 2.12 (br, 4H),
LC/MS: rt 2.44 min, m/z 575.1 (M + + 1).
試験例８
TRPCチャネルを一過的に発現したHEK293細胞内へのCa２＋の流入
（１）TRPCチャネルのHEK293細胞への導入および発現
　HEK293細胞（ヒト胎児腎由来細胞）にヒトTRPC3またはTRPC6遺伝子をリポフェクション法で導入し、細胞膜にTRPC6チャネルを発現させた。すなわち、TRPC6のプラスミドDNA（pCI-neo (promega社)）を、リポフェクション剤を用いてHEK293細胞へ導入後、37℃で24時間～48時間培養することで、TRPC6チャネルが細胞膜に発現したHEK293細胞（TRPC6発現HEK293細胞）を作製した。具体的には、非特許文献１（S. Sawamura et al., Mol. Pharmacol., 89, 348-363 (2016)）に記載された方法に従った。TRPC3遺伝子を使用し、同様にして、TRPC3チャネルが細胞膜に発現したHEK293細胞（TRPC3発現HEK293細胞）を作製した。 Test example 8
Influx of Ca 2+ into HEK293 cells that transiently express TRPC channels (1) Introduction and expression of TRPC channels into HEK293 cells Human TRPC3 or TRPC6 genes were introduced into HEK293 cells (human embryonic kidney-derived cells) by lipofection. TRPC6 channel was expressed in the cell membrane. That is, by introducing TRPC6 plasmid DNA (pCI-neo (Promega)) into HEK293 cells using a lipofection agent and culturing them at 37°C for 24 to 48 hours, HEK293 cells with TRPC6 channels expressed in their cell membranes were produced. (TRPC6-expressing HEK293 cells) were generated. Specifically, the method described in Non-Patent Document 1 (S. Sawamura et al., Mol. Pharmacol., 89, 348-363 (2016)) was followed. Using the TRPC3 gene, HEK293 cells in which the TRPC3 channel was expressed in the cell membrane (TRPC3-expressing HEK293 cells) were produced in the same manner.
（２）TRPC発現HEK293細胞内におけるカルシウムイオン濃度変化の測定方法
　TRPC3またはTRPC6チャネルを発現させたHEK293培養細胞をトリプシンで剥がし、カバーガラスに撒き直したのち、再び37℃で3時間培養した。カルシウム指示薬Fura-2 AM(終濃度1μM)が入った培地500μlにカバーガラスを移し替え、30分間37℃で培養し細胞に取り込ませた。細胞外溶液にはCa２＋含有溶液およびCa２＋フリー溶液を用意した。Ca２＋含有溶液：(2 mM CaCl2、132 mM NaCl、4 mM KCl、1mM MgCl2、5 mMグルコース。5 mM HEPES (pH7.4))、Ca２＋フリー溶液：(132 mM NaCl、4 mM KCl、1mM MgCl2、5 mMグルコース、5 mM HEPES (pH7.4))。 (2) Method for measuring changes in calcium ion concentration in HEK293 cells expressing TRPC Cultured HEK293 cells expressing TRPC3 or TRPC6 channels were detached with trypsin, plated on cover glasses, and cultured again at 37°C for 3 hours. The cover glass was transferred to 500 μl of a medium containing the calcium indicator Fura-2 AM (final concentration 1 μM) and incubated at 37° C. for 30 minutes to be incorporated into the cells. A Ca 2+ -containing solution and a Ca 2+ -free solution were prepared as extracellular solutions. Ca2 + -containing solution: (2mM CaCl2 , 132mM NaCl, 4mM KCl, 1mM MgCl2, 5mM glucose.5mM HEPES (pH7.4)), Ca2 + -free solution: (132mM NaCl, 4mM KCl , 1mM MgCl2 , 5mM glucose, 5mM HEPES (pH7.4)).
　Ca2+含有溶液に細胞を入れた後、室温で、Fura-2の蛍光強度（510 nm）をAQUACOSMOS（Hamamatsu Photonics社）で経時的に測定しつつ以下の操作を行った。
０分：Ca２＋含有溶液を入れた測定用チャンバーにTRPC3発現細胞またはTRPC6発現細胞を入れた。
２分後：Ca２＋含有溶液に所定量の試験化合物を添加した。
１０分後：測定終了 After placing the cells in a Ca 2+ -containing solution, the following operations were performed at room temperature while measuring the fluorescence intensity (510 nm) of Fura-2 over time using AQUACOSMOS (Hamamatsu Photonics).
0 minutes: TRPC3-expressing cells or TRPC6-expressing cells were placed in a measurement chamber containing a Ca 2+ -containing solution.
After 2 minutes: A predetermined amount of test compound was added to the Ca 2+ -containing solution.
10 minutes later: Measurement complete
試験例８－１
TRPC6チャネルを発現したHEK293細胞内へのCa２＋の流入に対する実施例化合物の作用
　チャネルを試験例８に記載の測定方法に従ってカルシウムイオン濃度変化を測定した。試験化合物にはPPZ2、L687、L-0042～L-0046（以下に示す化合物）を使用し、その添加量は1～30μMである。また、試験化合物に代えて0.1％のDMSOを使用し、対照とした。Fura-2の蛍光強度（Ratio）を経時的に測定することによって、TRPC6チャネル発現HEK293細胞内におけるCa２＋増加量（試験化合物添加後に測定された最大Ratio値から初期の（試験化合物添加前の）値を控除したΔRatio値）を求めた。DMSOを使用した対照よりもCa２＋増加量が大きかった試験化合物を、TRPC6チャネルを活性化する物質とした。 Test example 8-1
Effect of Example Compounds on Ca 2+ Influx into HEK293 Cells Expressing TRPC6 Channel Changes in calcium ion concentration of the channel were measured according to the measurement method described in Test Example 8. PPZ2, L687, L-0042 to L-0046 (compounds shown below) are used as test compounds, and the amount added is 1 to 30 μM. Additionally, 0.1% DMSO was used instead of the test compound as a control. By measuring the fluorescence intensity (Ratio) of Fura-2 over time, the increase in Ca 2+ in HEK293 cells expressing TRPC6 channel (from the maximum Ratio value measured after addition of the test compound to the initial value (before addition of the test compound)) was determined. ΔRatio value) was calculated. Test compounds that caused a greater Ca 2+ increase than the DMSO control were designated as substances that activate TRPC6 channels.
Figure JPOXMLDOC01-appb-C000030
 　L687およびPPZ2についての結果を図９に、および本発明の化合物についての結果を図１０に示す。PPZ2はTRPC6チャネルを活性化させ細胞内へCa２＋を流入させることが知られており、本試験でもPPZ2の添加により、濃度依存的なΔRatio値の増加が見られた。類似化合物であるL687（図９）やL-0042～L-0046（図１０）においても、濃度依存的に細胞内Ca２＋濃度を増加させたことから、これら化合物はTRPC6チャネルを活性化することが確認された。特にL-0044（実施例１化合物）において、最も強いTRPC6依存的な細胞内Ca２＋濃度増加が観察された。
Figure JPOXMLDOC01-appb-C000030
 The results for L687 and PPZ2 are shown in FIG. 9, and the results for the compounds of the invention are shown in FIG. 10. PPZ2 is known to activate TRPC6 channels and cause Ca 2+ to flow into cells, and in this study, addition of PPZ2 resulted in a concentration-dependent increase in the ΔRatio value. Similar compounds L687 (Figure 9) and L-0042 to L-0046 (Figure 10) also increased intracellular Ca 2+ concentration in a concentration-dependent manner, indicating that these compounds activate TRPC6 channels. was confirmed. In particular, the strongest TRPC6-dependent increase in intracellular Ca 2+ concentration was observed in L-0044 (Example 1 compound).
試験例８－２
TRPC3チャネルを発現したHEK293細胞内へのCa２＋の流入に対する実施例化合物の作用
　試験例８１に記載の方法で作製されたTRPC3発現HEK293細胞（TRPC3/HEK293）を使用し、試験化合物としてL-0042～L-0046（1-30μM）を使用した。試験例８－１と同様にして、各細胞内へのCa２＋増加量（ΔRatio）を求めた。
　得られた結果を図１１に示す。ヒトTRPC3チャネルを発現させたHEK293細胞において、本発明の化合物L-0042～L-0046は、濃度依存的に細胞内Ca2+濃度を増加させたことから、これら化合物はTRPC3チャネルも活性化することが確認された。特にL-0043（実施例2化合物）において、最も強いTRPC3依存的な細胞内Ca２＋濃度増加が観察された。 Test example 8-2
Effect of example compounds on Ca 2+ influx into HEK293 cells expressing TRPC3 channels TRPC3-expressing HEK293 cells (TRPC3/HEK293) prepared by the method described in Test Example 81 were used, and L-0042 was used as the test compound. ~L-0046 (1-30μM) was used. The amount of Ca 2+ increase (ΔRatio) into each cell was determined in the same manner as Test Example 8-1.
The results obtained are shown in FIG. In HEK293 cells expressing human TRPC3 channels, compounds L-0042 to L-0046 of the present invention increased intracellular Ca 2+ concentration in a concentration-dependent manner, indicating that these compounds also activate TRPC3 channels. This was confirmed. In particular, the strongest TRPC3-dependent increase in intracellular Ca 2+ concentration was observed in L-0043 (Example 2 compound).
試験例９
インビトロ心肥大応答に対するL-0044の抑制効果
　新生児ラット心筋細胞を3μM L-0044（実施例１化合物）とともに、あるいはL-0044なしで、1時間プレインキュベートした。その後、細胞を100nMエンドセリン－１（ET-1）で24時間刺激した。細胞サイズを測定するため、細胞を抗アクチニン抗体で染色した。
　得られた結果を図１２に示す。図１２は、ET-1投与が心筋細胞の肥大を誘導し、他方、L-0044の前処置はET-1を介した心肥大を減少させることを示している。これにより、L-0044単独で弱い心筋肥大が起こる傾向があるものの、エンドセリン－１刺激による病的な心肥大を、3μM L-0044での処置が抑制することが判明した。 Test example 9
Suppressive effect of L-0044 on in vitro cardiac hypertrophic response Neonatal rat cardiomyocytes were preincubated with or without 3 μM L-0044 (Example 1 compound) for 1 hour. Cells were then stimulated with 100 nM endothelin-1 (ET-1) for 24 hours. To measure cell size, cells were stained with anti-actinin antibody.
The obtained results are shown in FIG. 12. Figure 12 shows that ET-1 administration induces cardiomyocyte hypertrophy, while L-0044 pretreatment reduces ET-1-mediated cardiac hypertrophy. This revealed that although L-0044 alone tends to cause weak myocardial hypertrophy, treatment with 3 μM L-0044 suppresses pathological cardiac hypertrophy caused by endothelin-1 stimulation.
試験例１０
TRPC6チャネルを発現したHEK293細胞内へのZn２＋の流入に対する実施例化合物の作用
　TRPC6発現HEK293細胞（TRPC6-mCherryを発現するHEK293）もしくは対照HEK293細胞を使用し、試験化合物としてL-0043、L-0044、L-0046 (10μM)もしくは PPZ2 (30μM)を使用した。試験例５と同様にして、各細胞内へのZn2+増加量を求めた。得られた結果を図１３に示す。図１３はL-0043とL-0044はPPZ2よりも低濃度かつ強力にTRPC6を介してZn2+流入を増強することを示している。 Test example 10
Effect of Example Compounds on Zn 2+ Influx into HEK293 Cells Expressing TRPC6 Channel Using TRPC6-expressing HEK293 cells (HEK293 expressing TRPC6-mCherry) or control HEK293 cells, L-0043, L- 0044, L-0046 (10 μM) or PPZ2 (30 μM) were used. In the same manner as Test Example 5, the amount of increase in Zn 2+ into each cell was determined. The obtained results are shown in FIG. 13. Figure 13 shows that L-0043 and L-0044 potently enhance Zn 2+ influx through TRPC6 at lower concentrations than PPZ2.
試験例１１
　新生児ラット心筋細胞をL-0044(1μM)で30分プレインキュベートした。ハイスピードカメラ（60 fps）を搭載した顕微鏡に細胞を設置し、細胞の動きを動画撮影した。細胞を3分間電気刺激（5 V, 1 Hz, パルス幅10ミリ秒）した後にノルエピネフリン（NE） (0.1μM)で刺激し、細胞の動きの変化を評価した。心筋細胞の収縮力はImageJのプラグインであるMUSCLEMOTION（A Versatile Open Software Tool to Quantify Cardiomyocyte and Cardiac Muscle Contraction In Vitro and In Vivo, https://doi.org/10.1161/CIRCRESAHA.117.312067）で解析した。得られた結果を図１４に示す。図１４はL-0044前処置によってNE刺激による心筋細胞の収縮力が増強することを示している。 Test example 11
Neonatal rat cardiomyocytes were preincubated with L-0044 (1 μM) for 30 min. The cells were placed in a microscope equipped with a high-speed camera (60 fps) and videos of cell movement were taken. Cells were electrically stimulated for 3 minutes (5 V, 1 Hz, pulse width 10 ms) and then stimulated with norepinephrine (NE) (0.1 μM) to evaluate changes in cell movement. Cardiomyocyte contractility was analyzed using the ImageJ plug-in MUSCLEMOTION (A Versatile Open Software Tool to Quantify Cardiomyocyte and Cardiac Muscle Contraction In Vitro and In Vivo, https://doi.org/10.1161/CIRCRESAHA.117.312067). The obtained results are shown in FIG. 14. Figure 14 shows that L-0044 pretreatment enhances the contractile force of cardiac myocytes induced by NE stimulation.

Claims (11)

  1. 　TRPC3/6/7チャネル活性化物質を含有する、心不全または骨格筋不全を予防および／または処置するための医薬組成物。 A pharmaceutical composition for preventing and/or treating heart failure or skeletal muscle failure, containing a TRPC3/6/7 channel activator.
  2. 　TRPC3/6/7チャネル活性化物質が、式（１）：
    Figure JPOXMLDOC01-appb-C000001
     ［式中、
    　Ａは、炭素原子または窒素原子を示し、
    　Ｂは、炭素原子または窒素原子を示し、
    　Ｒ１は、置換されてもよいアルキル基を示し、複数あるときは同一または異なってよく、
    　ｍは、０～５の整数を示し、
    　Ｒ２は、ハロゲン原子；トリハロゲノメチル基；アルキル基；アシルオキシ基；アルコキシカルボニル基；置換されてもよいアミド基；置換されてもよいピペリジニルカルボニル基；或いは置換されてもよいピロリジニルカルボニル基を示し、複数あるときは同一または異なってよく、
    　ｎは、０～５の整数を示し、
    　ｐは、０または１を示し、そして
    　ｑは、０または１を示す。］
    で表される化合物、ハイパーフォリン、GSK1702934A、およびカンナビジオール；
    からなる群から選択される少なくとも１種の化合物である、請求項１記載の医薬組成物。     The TRPC3/6/7 channel activator has the formula (1):
    Figure JPOXMLDOC01-appb-C000001
     [In the formula,
    A represents a carbon atom or a nitrogen atom,
    B represents a carbon atom or a nitrogen atom,
    R 1 represents an optionally substituted alkyl group, and when there is a plurality of them, they may be the same or different,
    m represents an integer from 0 to 5,
    R 2 is a halogen atom; a trihalogenomethyl group; an alkyl group; an acyloxy group; an alkoxycarbonyl group; an optionally substituted amide group; an optionally substituted piperidinylcarbonyl group; or an optionally substituted pyrrolidinyl Indicates a carbonyl group, and when there are multiple groups, they may be the same or different,
    n represents an integer from 0 to 5,
    p represents 0 or 1, and q represents 0 or 1. ]
    Compounds represented by, hyperforin, GSK1702934A, and cannabidiol;
    The pharmaceutical composition according to claim 1, which is at least one compound selected from the group consisting of.
  3. 　TRPC3/6/7チャネル活性化物質が、式（１）：
    Figure JPOXMLDOC01-appb-C000002
     ［式中、
    　Ａは、炭素原子または窒素原子を示し、
    　Ｂは、炭素原子または窒素原子を示し、
    　Ｒ１は、ハロゲン原子、アルキル基、アルコキシ基、アシルオキシ基、アルコキシカルボニル基、またはハロゲン原子で置換されたアルキル基を示し、複数あるときは同一または異なってよく、
    　ｍは、０～５の整数を示し、
    　Ｒ２は、ハロゲン原子；トリハロゲノメチル基；アルキル基；アシルオキシ基；アルコキシカルボニル基；アルキル基、アルコキシカルボニル基およびハロゲン原子からなる群から選択される１種または２種の基で置換されてもよいアミド基；アルキル基、アルコキシカルボニル基およびハロゲン原子からなる群から選択される１種または２種以上の基で置換されてもよいピペリジニルカルボニル基；或いはアルキル基、アルコキシカルボニル基およびハロゲン原子からなる群から選択される１種または２種以上の基で置換されてもよいピロリジニルカルボニル基を示し、複数あるときは同一または異なってよく、
    　ｎは、０～５の整数を示し、
    　ｐは、０または１を示し、そして
    　ｑは、０または１を示す。］
    で表される化合物、ハイパーフォリン、GSK1702934A、およびカンナビジオール；
    からなる群から選択される少なくとも１種の化合物である、請求項２記載の医薬組成物。     The TRPC3/6/7 channel activator has the formula (1):
    Figure JPOXMLDOC01-appb-C000002
     [In the formula,
    A represents a carbon atom or a nitrogen atom,
    B represents a carbon atom or a nitrogen atom,
    R 1 represents a halogen atom, an alkyl group, an alkoxy group, an acyloxy group, an alkoxycarbonyl group, or an alkyl group substituted with a halogen atom, and when there is more than one, they may be the same or different;
    m represents an integer from 0 to 5,
    R 2 may be substituted with one or two groups selected from the group consisting of a halogen atom; a trihalogenomethyl group; an alkyl group; an acyloxy group; an alkoxycarbonyl group; an alkyl group, an alkoxycarbonyl group, and a halogen atom. A good amide group; a piperidinylcarbonyl group which may be substituted with one or more groups selected from the group consisting of an alkyl group, an alkoxycarbonyl group, and a halogen atom; or an alkyl group, an alkoxycarbonyl group, and a halogen atom Indicates a pyrrolidinylcarbonyl group which may be substituted with one or more groups selected from the group consisting of, and when there are multiple groups, they may be the same or different,
    n represents an integer from 0 to 5,
    p represents 0 or 1, and q represents 0 or 1. ]
    Compounds represented by, hyperforin, GSK1702934A, and cannabidiol;
    The pharmaceutical composition according to claim 2, which is at least one compound selected from the group consisting of.
  4. 　前記TRPC3/6/7チャネル活性化物質が、前記式（１）で表わされる化合物であって、
    　Ａは、炭素原子または窒素原子を示し、
    　Ｂは、炭素原子または窒素原子を示し、
    　Ｒ１は、フッ素原子、塩素原子、臭素原子、Ｃ１～Ｃ４アルキル基、Ｃ１～Ｃ４アルコキシ基、Ｃ２～Ｃ５アシルオキシ基、Ｃ２～Ｃ５アルコキシカルボニル基、またはトリハロゲノメチル基を示し、複数あるときは同一または異なってよく、
    　ｍは、０～４の整数を示し、
    　Ｒ２は、フッ素原子；塩素原子；臭素原子；トリフルオロメチル基；トリクロロメチル基；トリブロモメチル基；Ｃ１～Ｃ４アルキル基；Ｃ２～Ｃ５アシルオキシ基；Ｃ２～Ｃ５アルコキシカルボニル基；Ｃ１～Ｃ３アルキル基およびＣ２～Ｃ４アルコキシカルボニル基からなる群から選択される１種または２種の基で置換されてもよいアミド基；Ｃ１～Ｃ３アルキル基、Ｃ２～Ｃ４アルコキシカルボニル基およびハロゲン原子からなる群から選択される１種または２種以上の基で置換されてもよいピペリジニルカルボニル基；或いはＣ１～Ｃ３アルキル基、Ｃ２～Ｃ４アルコキシカルボニル基、およびハロゲン原子からなる群から選択される１種または２種以上の基で置換されてもよいピロリジニルカルボニル基を示し、複数あるときは同一または異なってよく、
    　ｎは、０～４の整数を示し、
    　ｐは、０または１を示し、そして
    　ｑは、０または１を示す、
    請求項３に記載の医薬組成物。     The TRPC3/6/7 channel activator is a compound represented by the formula (1),
    A represents a carbon atom or a nitrogen atom,
    B represents a carbon atom or a nitrogen atom,
    R 1 represents a fluorine atom, a chlorine atom, a bromine atom, a C1-C4 alkyl group, a C1-C4 alkoxy group, a C2-C5 acyloxy group, a C2-C5 alkoxycarbonyl group, or a trihalogenomethyl group, and when there is more than one, may be the same or different;
    m represents an integer from 0 to 4,
    R 2 is a fluorine atom; a chlorine atom; a bromine atom; a trifluoromethyl group; a trichloromethyl group; a tribromomethyl group; a C1-C4 alkyl group; a C2-C5 acyloxy group; a C2-C5 alkoxycarbonyl group; a C1-C3 alkyl and an amide group which may be substituted with one or two groups selected from the group consisting of a C2-C4 alkoxycarbonyl group; a C1-C3 alkyl group, a C2-C4 alkoxycarbonyl group, and a halogen atom; a piperidinylcarbonyl group which may be substituted with one or more selected groups; or one selected from the group consisting of a C1-C3 alkyl group, a C2-C4 alkoxycarbonyl group, and a halogen atom; Indicates a pyrrolidinylcarbonyl group that may be substituted with two or more types of groups, and when there are multiple groups, they may be the same or different,
    n represents an integer from 0 to 4,
    p represents 0 or 1, and q represents 0 or 1,
    The pharmaceutical composition according to claim 3.
  5. 　前記TRPC3/6/7チャネル活性化物質が、前記式（１）で表わされる化合物であって、
    　Ａは、窒素原子を示し、
    　Ｂは、炭素原子または窒素原子を示し、
    　Ｒ１は、フッ素原子、塩素原子、Ｃ１～Ｃ４アルキル基、Ｃ１～Ｃ４アルコキシ基、Ｃ２～Ｃ４アルコキシカルボニル基、トリフルオロメチル基、トリクロロメチル基、またはトリブロモメチル基を示し、複数あるときは同一または異なってよく、
    　ｍは、０～３の整数を示し、
    　Ｒ２は、フッ素原子；塩素原子；トリフルオロメチル基；トリクロロメチル基；Ｃ１～Ｃ４アルキル基；Ｃ２～Ｃ４アシルオキシ基；Ｃ２～Ｃ４アルコキシカルボニル基；Ｃ１～Ｃ３アルキル基およびＣ２～Ｃ４アルコキシカルボニル基からなる群から選択される１種または２種の基で置換されてもよいアミド基；Ｃ１～Ｃ３アルキル基、Ｃ２～Ｃ４アルコキシカルボニル基およびハロゲン原子からなる群から選択される１種または２種以上の基で置換されてもよいピペリジニルカルボニル基；或いはＣ１～Ｃ３アルキル基、Ｃ２～Ｃ４アルコキシカルボニル基およびハロゲン原子からなる群から選択される１種または２種以上の基で置換されてもよいピロリジニルカルボニル基を示し、複数あるときは同一または異なってよく、
    　ｎは、０～３の整数を示し、
    　ｐは、１を示し、そして
    　ｑは、１を示す、
    請求項４に記載の医薬組成物。     The TRPC3/6/7 channel activator is a compound represented by the formula (1),
    A represents a nitrogen atom,
    B represents a carbon atom or a nitrogen atom,
    R 1 represents a fluorine atom, a chlorine atom, a C1-C4 alkyl group, a C1-C4 alkoxy group, a C2-C4 alkoxycarbonyl group, a trifluoromethyl group, a trichloromethyl group, or a tribromomethyl group, and when there is more than one, may be the same or different;
    m represents an integer from 0 to 3,
    R 2 is a fluorine atom; a chlorine atom; a trifluoromethyl group; a trichloromethyl group; a C1 to C4 alkyl group; a C2 to C4 acyloxy group; a C2 to C4 alkoxycarbonyl group; a C1 to C3 alkyl group and a C2 to C4 alkoxycarbonyl group An amide group which may be substituted with one or two groups selected from the group consisting of; one or two groups selected from the group consisting of a C1-C3 alkyl group, a C2-C4 alkoxycarbonyl group, and a halogen atom; A piperidinylcarbonyl group which may be substituted with any of the above groups; or one or more groups selected from the group consisting of a C1-C3 alkyl group, a C2-C4 alkoxycarbonyl group, and a halogen atom. Indicates a pyrrolidinyl carbonyl group, which may be the same or different when there are multiple groups,
    n represents an integer from 0 to 3;
    p represents 1, and q represents 1,
    The pharmaceutical composition according to claim 4.
  6. 　前記TRPC3/6/7チャネル活性化物質が、前記式（１）で表わされる化合物であって、
    　Ａは、窒素原子を示し、
    　Ｂは、炭素原子または窒素原子を示し、
    　Ｒ１は、塩素原子、Ｃ１～Ｃ３アルキル基、Ｃ１～Ｃ３アルコキシ基、Ｃ２～Ｃ４アルコキシカルボニル基、トリフルオロメチル基、またはトリクロロメチル基を示し、複数あるときは同一または異なってよく、
    　ｍは、０～２の整数を示し、
    　Ｒ２は、フッ素原子；塩素原子；トリフルオロメチル基；トリクロロメチル基；Ｃ１～Ｃ４アルキル基；Ｃ２～Ｃ４アルコキシカルボニル基；Ｃ１～Ｃ３アルキル基およびＣ２～Ｃ４アルコキシカルボニル基からなる群から選択される１種または２種の基で置換されてもよいアミド基；或いはＣ１～Ｃ３アルキル基、Ｃ２～Ｃ４アルコキシカルボニル基およびハロゲン原子からなる群から選択される１種または２種以上の基で置換されてもよいピペリジニルカルボニル基を示し、複数あるときは同一または異なってよく、
    　ｎは、０～３の整数を示し、
    　ｐは、１を示し、
    　ｑは、１を示す、
    請求項５記載の医薬組成物。     The TRPC3/6/7 channel activator is a compound represented by the formula (1),
    A represents a nitrogen atom,
    B represents a carbon atom or a nitrogen atom,
    R 1 represents a chlorine atom, a C1-C3 alkyl group, a C1-C3 alkoxy group, a C2-C4 alkoxycarbonyl group, a trifluoromethyl group, or a trichloromethyl group, and when there is more than one, they may be the same or different,
    m represents an integer from 0 to 2,
    R 2 is selected from the group consisting of a fluorine atom; a chlorine atom; a trifluoromethyl group; a trichloromethyl group; a C1-C4 alkyl group; a C2-C4 alkoxycarbonyl group; a C1-C3 alkyl group and a C2-C4 alkoxycarbonyl group. an amide group which may be substituted with one or two groups; or substituted with one or more groups selected from the group consisting of a C1-C3 alkyl group, a C2-C4 alkoxycarbonyl group, and a halogen atom; Indicates a piperidinyl carbonyl group that may be
    n represents an integer from 0 to 3;
    p indicates 1,
    q indicates 1,
    The pharmaceutical composition according to claim 5.
  7. 　前記TRPC3/6/7チャネル活性化物質が、以下に示された化合物の１種または２種以上である、請求項１記載の医薬組成物：
    Figure JPOXMLDOC01-appb-C000003
    Figure JPOXMLDOC01-appb-C000004
    Figure JPOXMLDOC01-appb-C000005
         The pharmaceutical composition according to claim 1, wherein the TRPC3/6/7 channel activator is one or more of the following compounds:
    Figure JPOXMLDOC01-appb-C000003
    Figure JPOXMLDOC01-appb-C000004
    Figure JPOXMLDOC01-appb-C000005
  8. 　心不全が、急性心不全、または慢性心不全における急性増悪である、請求項１記載の医薬組成物。 The pharmaceutical composition according to claim 1, wherein the heart failure is acute heart failure or acute exacerbation of chronic heart failure.
  9. 　心不全または骨格筋不全を予防および／または処置するための物質をスクリーニングする方法であって、TRPC3/6/7チャネル活性化物質を選別する、該スクリーニング方法。 A method of screening for substances for preventing and/or treating heart failure or skeletal muscle failure, the screening method comprising selecting TRPC3/6/7 channel activators.
  10. 　被験物質の存在下で、細胞中の亜鉛濃度を測定する工程、および前記亜鉛濃度が、前記被験物質の非存在下における亜鉛濃度と比較して増加していた場合に、前記被験物質はTRPC3/6/7チャネル活性化をもたらす候補物質であると判断する工程
    を備える、請求項８記載のスクリーニング方法。     measuring the zinc concentration in cells in the presence of a test substance, and if the zinc concentration is increased compared to the zinc concentration in the absence of the test substance, the test substance 9. The screening method according to claim 8, comprising the step of determining that the substance is a candidate substance that causes 6/7 channel activation.
  11. 　式（１）：
    Figure JPOXMLDOC01-appb-C000006
     ［式中、
    　Ａは、炭素原子または窒素原子を示し、
    　Ｂは、炭素原子または窒素原子を示し、
    　Ｒ１は、ハロゲン原子、アルキル基、アルコキシ基、アシルオキシ基、アルコキシカルボニル基、またはハロゲン原子で置換されたアルキル基を示し、複数あるときは同一または異なってよく、
    　ｍは、０～５の整数を示し、
    　ｎは、２～５の整数を示し、
    　複数あるＲ２のうち、少なくとも１つはハロゲン原子、アルキル基またはトリフルオロメチル基であり；残りのＲ２は、１）単数または複数のハロゲン原子で置換され、かつアルキル基およびアルコキシカルボニル基からなる群から選択される１種または２種以上の基で置換されてもよいピペリジニルカルボニル基；或いは２）単数または複数のハロゲン原子で置換され、かつアルキル基およびアルコキシカルボニル基からなる群から選択される１種または２種以上の基で置換されてもよいピロリジニルカルボニル基を示し、複数あるときは同一または異なってよく、
    　ｐは、０または１を示し、そして
    　ｑは、０または１を示す。］
    で表される化合物。     Formula (1):
    Figure JPOXMLDOC01-appb-C000006
     [In the formula,
    A represents a carbon atom or a nitrogen atom,
    B represents a carbon atom or a nitrogen atom,
    R 1 represents a halogen atom, an alkyl group, an alkoxy group, an acyloxy group, an alkoxycarbonyl group, or an alkyl group substituted with a halogen atom, and when there is more than one, they may be the same or different;
    m represents an integer from 0 to 5,
    n represents an integer from 2 to 5,
    At least one of the plurality of R 2 is a halogen atom, an alkyl group, or a trifluoromethyl group; the remaining R 2 are 1) substituted with a single or multiple halogen atoms, and are selected from an alkyl group and an alkoxycarbonyl group. or 2) a piperidinylcarbonyl group optionally substituted with one or more groups selected from the group consisting of alkyl groups and alkoxycarbonyl groups; or 2) substituted with one or more halogen atoms; Indicates a pyrrolidinyl carbonyl group that may be substituted with one or more selected groups, and when there are multiple groups, they may be the same or different,
    p represents 0 or 1, and q represents 0 or 1. ]
    A compound represented by
PCT/JP2023/030211 2022-08-23 2023-08-22 Heart failure treatment via cardiotonic effect by trpc3/6/7 channel activation WO2024043242A1 (en)

Applications Claiming Priority (2)

Application Number Priority Date Filing Date Title
JP2022132533 2022-08-23
JP2022-132533 2022-08-23

Publications (1)

Publication Number Publication Date
WO2024043242A1 true WO2024043242A1 (en) 2024-02-29

Family

ID=90013414

Family Applications (1)

Application Number Title Priority Date Filing Date
PCT/JP2023/030211 WO2024043242A1 (en) 2022-08-23 2023-08-22 Heart failure treatment via cardiotonic effect by trpc3/6/7 channel activation

Country Status (1)

Country Link
WO (1) WO2024043242A1 (en)

Citations (4)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
JP2002520260A (en) * 1998-07-09 2002-07-09 シーヴイ テクノロジーズ インコーポレイテッド Hypericin and hypericum extract: specific T-type calcium channel blockers and their use as therapeutics targeting T-type calcium channels
JP2018502823A (en) * 2014-10-14 2018-02-01 ジーダブリュー・ファーマ・リミテッドＧｗ Ｐｈａｒｍａ Ｌｉｍｉｔｅｄ Use of cannabinoids in the treatment of degenerative skeletal muscle disease
WO2021165992A1 (en) * 2020-02-19 2021-08-26 DR. MERCHANT, Shreema Compositions and therapeutic uses of cannabidiol
WO2022118966A1 (en) * 2020-12-04 2022-06-09 国立大学法人京都大学 Agent for promoting nucleic acid molecule uptake into cells, pharmaceutical composition, and novel compound

Patent Citations (4)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
JP2002520260A (en) * 1998-07-09 2002-07-09 シーヴイ テクノロジーズ インコーポレイテッド Hypericin and hypericum extract: specific T-type calcium channel blockers and their use as therapeutics targeting T-type calcium channels
JP2018502823A (en) * 2014-10-14 2018-02-01 ジーダブリュー・ファーマ・リミテッドＧｗ Ｐｈａｒｍａ Ｌｉｍｉｔｅｄ Use of cannabinoids in the treatment of degenerative skeletal muscle disease
WO2021165992A1 (en) * 2020-02-19 2021-08-26 DR. MERCHANT, Shreema Compositions and therapeutic uses of cannabidiol
WO2022118966A1 (en) * 2020-12-04 2022-06-09 国立大学法人京都大学 Agent for promoting nucleic acid molecule uptake into cells, pharmaceutical composition, and novel compound

Non-Patent Citations (2)

* Cited by examiner, † Cited by third party
Title
ODA SAYAKA, NISHIYAMA KAZUHIRO, FURUMOTO YUKA, YAMAGUCHI YOHEI, NISHIMURA AKIYUKI, TANG XIAOKANG, KATO YURI, NUMAGA-TOMITA TAKURO,: "Myocardial TRPC6-mediated Zn2+ influx induces beneficial positive inotropy through β-adrenoceptors", NATURE COMMUNICATIONS, NATURE PUBLISHING GROUP, UK, vol. 13, no. 1, 26 October 2022 (2022-10-26), UK, XP093143545, ISSN: 2041-1723, DOI: 10.1038/s41467-022-34194-9 *
OKUBO REIMA, ODA SAYAKA, NISHIYAMA KAZUHIRO, NISHIMURA AKIYUKI, KATO YURI, MI XINYA, NISHIDA MOTOHIRO: "O4-1 Regulation of cardiac baroreflex response by TRPC6-mediated Zn2+ entry", FORUM 2022 PHARMACEUTICAL HEALTH SCIENCES: ENVIRONMENTAL TOXICOLOGY, LECTURE ABSTRACTS; AUGUST 30-31, 2022, 17 August 2022 (2022-08-17) - 31 August 2022 (2022-08-31), pages 158, XP009554627 *

Similar Documents

Publication Publication Date Title
ES2707596T3 (en) Use of an adrenal hormone-modifying agent
AU2014254052B2 (en) Treatment of cancer with dihydropyrazino-pyrazines
TW201501711A (en) Pyrazole-amide compounds and medical use thereof
TW201031662A (en) Imidazolylpyrazine derivatives
JP2015504923A (en) Combination therapy using late sodium ion channel blockers and potassium ion channel blockers
JP2021500366A (en) Substituted imidazopyridine amides and their use
WO2020254289A1 (en) N-(phenyl)-indole-3-sulfonamide derivatives and related compounds as gpr17 modulators for treating cns disorders such as multiple sclerosis
CA2812061A1 (en) Modulators of the gpr119 receptor and the treatment of disorders related thereto
CA2974958A1 (en) Compositions and methods of using tyrosine kinase inhibitors
KR20240127394A (en) Treatment of brain tumors and neuroblastomas
WO2024043242A1 (en) Heart failure treatment via cardiotonic effect by trpc3/6/7 channel activation
TW201536748A (en) Fluorene-amide compounds and pharmaceutical use thereof
WO2019117812A1 (en) Near infra-red molecular probes for use in diagnosis of fibrotic conditions and screening of anti-fibrotic drugs
WO2001055093A1 (en) N-arylhydrazide compounds and use thereof as drugs
BR112020007983A2 (en) prodrugs of substituted triazole derivatives and uses thereof
EP4013497B1 (en) Compounds suitable for the treatment and prophylaxis of muscle wasting and other conditions
WO2018208848A1 (en) Compounds and methods for treatment of visceral pain
WO2021123297A1 (en) Oga inhibitor compounds
TW202133842A (en) Combination of iap inhibitors and parp or mek inhibitors or other chemotherapeutic agents
WO2020068846A1 (en) Heterocyclic compound
WO2018208847A1 (en) Compounds and methods for treatment of pain from inflammatory bowel disease
JP6921100B2 (en) Heterocyclic compound
EP3881840A1 (en) Sortilin antagonists for use inthe treatment of diabetic retinopathy
WO2024089691A1 (en) Modulators and uses thereof
KR20230095193A (en) EGFR and HDAC dual inhibitor compounds and medical use thereof

Legal Events

Date Code Title Description
121 Ep: the epo has been informed by wipo that ep was designated in this application

Ref document number: 23857351

Country of ref document: EP

Kind code of ref document: A1

DPE1 Request for preliminary examination filed after expiration of 19th month from priority date (pct application filed from 20040101)