JPWO2019082940A1 - Ryanodine receptor inhibitor - Google Patents

Abstract

To provide a medicine containing a compound having ryanodine inhibitory activity, particularly RyR1 inhibitory activity, as an active ingredient. Ryanodine receptor-related with a compound represented by the general formula (I) (in the formula, all symbols have the same meaning as described herein) or a pharmaceutically acceptable salt thereof as an active ingredient. Treatment or preventive drug for the disease.

Description

The present invention relates to ryanodine receptor inhibitors.

The ryanodine receptor (RyR) is a Ca ²⁺ release channel responsible for the release of calcium ions (Ca ²⁺ ) from the sarcoplasmic reticulum (SR), an essential step for muscle contraction. RyR is stimulated by Ca ²⁺ on the cytosol side to release Ca ²⁺ into the cytosol (Ca ²⁺ -induced Ca ²⁺ release, CICR). Therefore, it acts as a positive feedback mechanism, and a small amount of Ca ²⁺ in the cytosol near the channel causes more Ca ²⁺ release from SR.

There are known subtypes of RyR: RyR1, which is mainly expressed in skeletal muscle, RyR2, which is mainly expressed in myocardium, and RyR3, which is widely expressed but is highly expressed in the brain. RyR gene mutation causes abnormal increase in CICR activity and is considered to be a cause of various diseases. For example, RyR1-related diseases include malignant hyperthermia (MH) (for example, increased body temperature due to inhalation anesthetic during surgery, skeletal muscle contracture, etc.), central core disease (CCD) (muscle strength of limbs designated as intractable diseases) (Decrease, etc.) (Non-Patent Document 1). RyR2-related diseases include catecholaminergic polymorphic ventricular tachycardia (CPVT) (motor arrhythmia), arrhythmogenic right ventricular cardiomyopathy (ARVC) (right ventricular dysplasia, arrhythmia induction), and idiopathic ventricular. Examples thereof include arrhythmia (IVF) (inducing ventricular fibrillation at rest) (Non-Patent Document 2). Furthermore, as diseases resulting from secondary dysregulation of RyR due to main factors other than RyR, malignant syndrome (side effect of schizophrenia drug), muscular dystrophy, Alzheimer's dementia, Huntington's disease, traumatic injury Examples include sexual brain injury (Non-Patent Documents 3 to 8).

The cause of these RyR-related diseases is considered to be an abnormal increase in CICR activity due to a mutation in the RyR gene, but there is currently no effective therapeutic agent. For example, dantrolene is the only therapeutic agent for malignant hyperthermia (Non-Patent Document 9).

Special Publication No. 51-18440 International Publication No. 2010/064701 Pamphlet

Orphanet J Rare Dis. 10:93 (2015) J. Electrocardiol. 48 (5), 874-878 (2015) Neuropsychiatr Dis Treat. 13: 161-175 (2017) FASEB J. 32: 1025-1043 (2018) J. Neurosci. 35: 6893-6902 (2015) J. Alzheimers Dis. 45: 561-580 (2015) Molecular Neurodegeneration 2011, 6:81, 1-12 Eur Spine J (2009) 18: 1442-1451 Traumatic brain injury Guidelines for Management of Malignant Hyperthermia Patients 2016-For Safe Anesthesia Management-August 2016 Created by Japan Anesthesiology Society Safety Committee Malignant Hyperthermia WG Hum Mutat., 37, 1231-1241, 2016 J. Med. Chem. 1980, 23, 1358-1363 Chem. Pharm. Bull. 41 (1) 126-131 (1993) Nat. Commun., 2014, 5: 4153

However, dantrolene is unsuitable for long-term administration due to its strong side effects, and its low water solubility limits its administration method. In addition, dantrolene is not applicable as a therapeutic agent for CCD. Therefore, an object of the present invention is to provide a drug that has ryanodine receptor inhibitory activity, particularly inhibitory activity on RyR1, and is effective for ryanodine receptor-related diseases.

Therefore, as a result of various studies in view of the relationship between the ryanodine receptor and CICR, which is an indispensable step for muscle contraction, the present inventors have found that cultured cells expressing the fluorescent endoplasmic reticulum Ca ²⁺ indicator and the disease mutant RyR. and fluorescent endoplasmic reticulum Ca ²⁺ indicator and the cultured cells expressing wild-type RyR cultured respectively, to measure the endoplasmic reticulum Ca ²⁺ concentration in the presence of a test substance, the endoplasmic reticulum Ca ²⁺ in both cultured cells It was found that a CICR activity inhibitor, that is, a therapeutic agent for a ryanodine receptor-related disease can be screened by comparing the concentrations, and a patent application was filed earlier (Japanese Patent Application No. 2016-13147, Non-Patent Document 10). Then, as a result of screening various compounds using this screening method, the compound represented by the following general formula (I) has excellent ryanodine receptor inhibitory activity, and is a prophylactic or therapeutic agent for ryanodine receptor-related diseases. The present invention was completed by finding that it is useful as a compound.

That is, the present invention provides the following [1] to [9].
[1] General formula (I)

[In the formula, R ¹ represents a C1-12 alkyl group, a C2-12 alkenyl group or a C2-12 alkynyl group, and the alkyl group, the alkenyl group and the alkynyl group are a C3-12 carbocycle or a 4- to 12-membered complex. The rings may be substituted,
X indicates CR ⁴ or N,
R ^{2, R 3} and ^{R 4} are each independently a hydrogen atom, Cl to 6 alkyl, C2-6 alkenyl, C2-6 alkynyl, C1 -3 fluoroalkyl group, Cl to 6 alkoxy group, OR ¹⁰ (in the formula, R ¹⁰ represents a hydrogen atom or a phenyl group), NR ¹¹ R ¹² (in the formula, R ¹¹ and R ¹² each independently represent a hydrogen atom, a C1-6 alkyl group or a phenyl group). ), COR ¹³ (in the formula, R ¹³ is a hydroxyl group, OR ²⁰ (in the formula, R ²⁰ represents a hydrogen atom, a C1-6 alkyl group or a phenyl group) or NR ²¹ R ²² (in the formula, R ²¹ and R ²² independently indicate a hydrogen atom, a C1-6 alkyl group or a phenyl group)), a cyano group, a nitro group, a halogen atom, a C1 to 3 fluoride alkoxy group, and a C1 to 6 alkylthio group. , C3-12 carbocycles or 4-12-membered heterocycles, and R ² and R ³ may together represent a C3-5 alkylene group or a C1-3 alkylenedioxy group.
Y is COR ³⁰ (R ³⁰ is a hydroxyl group, OR ⁴⁰ (in the formula, R ⁴⁰ indicates a C1-6 alkyl group or a phenyl group) or NR ⁴¹ R ⁴² (in the formula, R ⁴¹ and R ⁴² are independent of each other). , Hydrogen atom, C1-6 alkyl group or phenyl group.) Is shown.). ]
A therapeutic or prophylactic agent for a ryanodine receptor-related disease containing the compound represented by (1) or a pharmaceutically acceptable salt thereof as an active ingredient.

[2] In the compound represented by the general formula (I) described in the previous section [1], X is CH, R ¹ is a C6 to 10 alkyl group, R ² is a hydrogen atom, and R ³ is C 1. The therapeutic or prophylactic agent according to the preceding paragraph [1], which is a ~ 4 alkyl group.

[3] The therapeutic and / or prophylactic agent according to the preceding item [1] or [2], wherein the ryanodine receptor is RyR1.

[4] The therapeutic or prophylactic agent according to any one of the above items [1] to [3], wherein the ryanodine receptor-related disease is a disease selected from the group consisting of malignant hyperthermia and central core disease.

[5] An inhibitor of a ryanodine receptor containing a compound represented by the general formula (I) described in the preceding paragraph [1] or a pharmaceutically acceptable salt thereof as an active ingredient.

[6] The inhibitor according to the previous item [5], wherein the ryanodine receptor is RyR1.

[7] General formula (IA)

[In the formula, R ^1A represents a C1-12 alkyl group, a C3-6 cycloalkyl-C1-4 alkyl group or a phenyl-C1-4 alkyl group.
X ^A represents CR ^4A or N, R ^4A represents a hydrogen atom, a C1-4 alkyl group or a C1-4 alkoxy group.
One of R ^2A and R ^3A is a hydrogen atom, C1 to 4 alkyl group or C1 to 4 alkoxy group, and the other is hydrogen atom, C1 to 4 alkyl group, C1 to 4 alkoxy group, cyano group, halogen atom and trihalomethyl. A group, a trihalomethyloxy group or a methylthio group, or R ^2A and R ^3A together to indicate a C3-5 alkylene group or an -O-CH _2- O- group (provided that (i) R ^2A and When both R ^3A are hydrogen atoms, R ^1A shows a cyclohexylmethyl group,
(Ii) When R ^1A is an n-propyl group, R ^2A is not a methyl group or a methoxy group,
(Iii) R ^1A is not an octyl group when R ^2A and R ^3A together indicate an -O-CH _2- O- group)] or a pharmaceutically acceptable salt thereof.

[8] In the compound represented by the general formula (IA) described in the previous item [7],
One of R ^2A and R ^3A is a hydrogen atom and the other is a C1-4 alkyl group, a C1-4 alkoxy group, a halogen atom, a cyano group or a trihalomethyl group, or R ² and R ³ are combined. (However, (i) when R ^1A is an n-propyl group, R ^2A is not a methyl group or a methoxy group, but an -O-CH _2- O- group.
(Ii) R ^1A is not an octyl group when R ^2A and R ^3A together indicate an -O-CH _2- O- group)]
The compound according to claim 7 or a pharmaceutically acceptable salt thereof.

[9] (1) 1-Cyclohexylmethyl-1,4-dihydro-4-oxoquinoline-3-carboxylic acid,
(2) 1,4-dihydro-6-methyl-1-octyl-4-oxoquinoline-3-carboxylic acid,
(3) 1,4-Dihydro-7-methyl-1-propyl-4-oxoquinoline-3-carboxylic acid,
(4) 1,4-dihydro-7-methyl-1-octyl-4-oxoquinoline-3-carboxylic acid,
(5) 1,4-dihydro-7-methyl-1-octyl-4-oxoquinoline-3-carboxylic acid,
(6) 1,4-Dihydro-7-methoxy-4-oxo-1-propylquinoline-3-carboxylic acid,
(7) 1,4-Dihydro-7-methoxy-1-octyl-4-oxoquinoline-3-carboxylic acid,
(8) 1,4-Dihydro-1-propyl-4-oxo-7-trifluoromethylquinoline-3-carboxylic acid,
(9) 1,4-dihydro-1-octyl-4-oxo-7-trifluoromethylquinoline-3-carboxylic acid,
(10) 5-decyl-8-oxo- [1,3] dioxolo [4,5-g] quinoline-7-carboxylic acid,
(11) 1,4-Dihydro-7-methoxy-1-propyl-4-oxo-1,8-naphthylidine-3-carboxylic acid,
(12) 1,4-Dihydro-7-methoxy-1-octyl-4-oxo-1,8-naphthylidine-3-carboxylic acid,
(13) 1-decyl-1,4-dihydro-7-methoxy-4-oxo-1,8-naphthylidine-3-carboxylic acid,
(14) 1,4-dihydro-6,7-dimethoxy-1-octyl-4-oxoquinoline-3-carboxylic acid,
(15) 1-decyl-1,4-dihydro-6,7-dimethoxy-4-oxoquinoline-3-carboxylic acid,
(16) 1-propyl-4-oxo-cyclopenta [g] quinoline-3-carboxylic acid,
(17) 1-octyl-4-oxo-cyclopenta [g] quinoline-3-carboxylic acid,
(18) 1-propyl-1,4-dihydro-7-trifluoromethyloxy-4-oxoquinoline-3-carboxylic acid,
(19) 1-propyl-1,4-dihydro-7-trifluoromethyloxy-4-oxoquinoline-3-carboxylic acid,
(20) 1,4-dihydro-8-methoxy-1-octyl-4-oxoquinoline-3-carboxylic acid,
(21) 7,8-dimethoxy-1,4-dihydro-1-octyl-4-oxoquinoline-3-carboxylic acid, and (22) 1,4-dihydro-7-methylthio-1-octyl-4-oxo The compound according to the preceding item [7] or a pharmaceutically acceptable salt thereof selected from the group consisting of quinoline-3-carboxylic acid.

The compound of the present invention represented by the general formula (I) or a pharmaceutically acceptable salt thereof exhibits inhibitory activity on the ryanodine receptor, particularly the RyR1 receptor, and is therefore useful for the prevention or treatment of skeletal muscle disease.

It shows the RyR1 inhibitory activity of oxolinic acid and 1,4-dihydro-4-oxoquinoline 3-carboxylic acid (DOCA). The RyR1 inhibitory activity of the compound used in the present invention is shown. The RyR1 inhibitory activity of the compound used in the present invention is shown. The RyR1 inhibitory activity of the compound used in the present invention is shown. The RyR1 binding activity of the compound used in the present invention is shown. It shows a therapeutic effect on RyR1 malignant hyperthermia mutation-introduced mice.

The active ingredient in the therapeutic or prophylactic agent for the ryanodine receptor-related disease of the present invention is a compound represented by the general formula (I) or a pharmaceutically acceptable salt thereof. The compound represented by the general formula (I) includes an oxolinic acid and a quinoline-3-carboxylic acid derivative compound. Oxolinic acid is a quinolone-based antibacterial agent developed in the 1970s (Patent Document 1). Oxolinic acid inhibits bacterial DNA gyrase and is now used primarily as an antibacterial agent for animals. In addition, as a compound having a structure similar to that of oxolinic acid, a study on the structure-activity relationship regarding the antibacterial activity of a 1,4-dihydro-4-oxoquinoline-3-carboxylic acid (DOCA) derivative has been conducted (for example, non-patent). Documents 11 and 12). Although these compounds are known to have antibacterial activity, they are not known to have ryanodine receptor inhibitory activity. Moreover, the compound represented by the general formula (IA) is a novel compound which has not been known so far.
Hereinafter, the present invention will be described in detail.

In the present specification, the "C1-12 alkyl group" refers to a linear alkyl group having 1 to 12 carbon atoms or a branched-chain alkyl group having 3 to 12 carbon atoms. Examples of C1-12 alkyl groups include methyl group, ethyl group, n-propyl group, n-butyl group, n-pentyl group, n-hexyl group, n-heptyl group, n-octyl group and n-nonyl. Group, n-decyl group, n-undecyl group, n-dodecyl group, linear alkyl group, isopropyl group, isobutyl group, t-butyl group, s-butyl group, isopentyl group, 1-ethylpropyl group, 3 -Branch chains such as methylbutyl group, 2,2-dimethylpropyl group, isohexyl group, 3-ethylbutyl group, 3,3-dimethylbutyl group, isoheptenyl group, isooctyl group, isononyl group, isodecyl, isoundecyl group, isododecyl group, etc. Alkyl group in the form can be mentioned.

In the present specification, the “C2-12 alkenyl group” refers to a linear alkenyl group having 2 to 12 carbon atoms or a branched chain alkenyl group having 3 to 12 carbon atoms. Examples of C2-12 alkenyl groups are ethenyl group, n-propenyl group, n-butenyl group, n-pentenyl group, n-hexenyl group, n-heptanyl group, n-octanyl group, n-nonenyl group, n. -Desenyl group, n-undecenyl group, n-dodecenyl group, linear alkenyl group, isopropenyl group, isobutenyl group, s-butenyl group, isopentenyl group, 1-ethylpropenyl group, 3-methylbutenyl group, 2, Branched chain alkenyl groups such as 2-dimethylpropenyl group, isohexenyl group, 3-ethylbutenyl group, 3,3-dimethylbutenyl group, isoheptenyl group, isooctenyl group, isononenyl group, isodecenyl, isoundecenyl group, isododecenyl group Can be mentioned.

In the present specification, the “C2-12 alkynyl group” refers to a linear alkynyl group having 2 to 12 carbon atoms or a branched alkynyl group having 3 to 12 carbon atoms. Examples of C2-12 alkynyl groups include ethynyl group, n-propynyl group, n-butynyl group, n-pentynyl group, n-hexynyl group, n-heptinyl group, n-octynyl group, n-noninyl group, n. -Desinyl group, n-undecynyl group, n-dodecynyl group linear alkynyl group, isobutynyl group, s-butynyl group, isopentinyl group, 1-ethylpropynyl group, 3-methylbutynyl group, 2,2-dimethylpropynyl group , Isohexynyl group, 3-ethylbutynyl group, 3,3-dimethylbutynyl group, isoheptinyl group, isooctynyl group, isononinyl group, isodecynyl, isoundecynyl group, isododecynyl group and other branched alkynyl groups.

In the present specification, "C3-12 carbocycle" refers to saturated hydrocarbons (cycloalkane) such as cyclopropane, cyclobutene, cyclopentene, cyclohexane, cycloheptane, cyclooctane, cyclononane, and cyclodecane, bicycloheptane, bicyclooctane, and bicyclononan. Bicyclo rings such as cyclobutene, cyclopentene, cyclohexene, cycloheptene, cyclooctene, cyclononen, cyclodecene and the like, cycloalkenes such as benzene, naphthalene, indan, inden and the like, and completely unsaturated or incompletely unsaturated carbon rings. Can be mentioned.

In the present specification, the "4 to 12 membered heterocycle" includes a "4 to 12 member saturated heterocycle" and a "4 to 12 member unsaturated heterocycle".
A "4- to 12-membered saturated heterocycle" is a saturated monocyclic or bicyclic or higher heterocycle having a heteroatom selected from a nitrogen atom, an oxygen atom and a sulfur atom, for example, morpholin, 1-. Examples thereof include pyrrolidine, piperidine, piperazine, tetrahydrofuran, tetrahydropyran, tetrahydrothiophene, thiazolidine, oxazolidine and the like.

In the present specification, the "4- to 12-membered unsaturated heterocycle" is a completely unsaturated or partially unsaturated monocyclic or bicyclic ring having a heteroatom selected from a nitrogen atom, an oxygen atom and a sulfur atom. The above heterocycles, for example, imidazoline, thiophene, furan, pyrrole, oxazole, isoxazole group, thiazole, isothiazole group, thiadiazol group, pyrazole, triazole, tetrazole, pyridine, pyrazine, pyrimidine, pyridazine, indole, isoindole, Examples thereof include indazole group, triazolopyridine, benzoimidazole, benzoxazole, benzothiazole group, benzothiophene, benzofuran, purine, quinoline, isoquinoline, quinazoline, quinoxaline, methylenedioxybenzene, ethylenedioxybenzene, dihydrobenzofuran and the like.

In the present specification, the “C1 to 6 alkyl group” refers to a linear alkyl group having 1 to 6 carbon atoms or a branched chain alkyl group having 3 to 6 carbon atoms. Examples of C1-6 alkyl groups include methyl group, ethyl group, n-propyl group, n-butyl group, n-pentyl group, n-hexyl group, linear alkyl group, isopropyl group, isobutyl group, and the like. t-Butyl group, s-butyl group, isopentyl group, 1-ethylpropyl group, 3-methylbutyl group, 2,2-dimethylpropyl group, isohexyl group, 3-ethylbutyl group, 3,3-dimethylbutyl group, etc. A branched alkyl group of the above can be mentioned.

In the present specification, the “C1 to 4 alkyl group” refers to a linear alkyl group having 1 to 4 carbon atoms or a branched chain alkyl group having 3 to 4 carbon atoms. Examples of C1-4 alkyl groups include linear alkyl groups such as methyl group, ethyl group, n-propyl group and n-butyl group, isopropyl group, isobutyl group, t-butyl group and s-butyl group. Examples thereof include branched alkyl groups.

In the present specification, the "C2-6 alkenyl group" refers to a linear alkenyl group having 2 to 6 carbon atoms or a branched chain alkenyl group having 3 to 6 carbon atoms. Examples of C2-6 alkenyl groups include ethenyl group, n-propenyl group, n-butenyl group, n-pentenyl group, n-hexenyl group, linear alkenyl group, isopropenyl group, isobutenyl group, s- Examples thereof include branched alkenyl groups such as a butenyl group, an isopentenyl group, a 1-ethylpropenyl group, a 3-methylbutenyl group and a 2,2-dimethylpropenyl group.

In the present specification, the “C2-6 alkynyl group” refers to a linear alkynyl group having 2 to 6 carbon atoms or a branched alkynyl group having 3 to 6 carbon atoms. Examples of C2-6 alkynyl groups include ethynyl group, n-propynyl group, n-butynyl group, n-pentynyl group, n-hexynyl group, linear alkynyl group, isobutynyl group, s-butynyl group, isopentynyl. Examples thereof include a branched alkynyl group such as a group, a 1-ethylpropynyl group, a 3-methylbutynyl group, a 2,2-dimethylpropynyl group and an isohexynyl group.

In the present specification, "C1 to 3 alkyl fluoride groups" include fluoromethyl, difluoromethyl, trifluoromethyl, fluoroethyl, difluoroethyl, trifluoroethyl, fluoropropyl, difluoropropyl, trifluoropropyl, and hexafluoropropyl. Group etc. can be mentioned.

In the present specification, the “C1 to 6 alkoxy group” refers to a linear alkyloxy group having 1 to 6 carbon atoms or a branched chain alkyloxy group having 3 to 6 carbon atoms. Examples of the C1-6 alkoxy group include methyloxy, ethyloxy, propyloxy, butyloxy, pentyloxy, hexyloxy group and the like.

In the present specification, examples of the "C1-4 alkoxy group" include methyloxy, ethyloxy, propyloxy, butyloxy group and the like.

In the present specification, "C1-6 alkylthio group" refers to a linear alkylthio group having 1 to 6 carbon atoms or a branched chain alkylthio group having 3 to 6 carbon atoms. Examples of the C1-6 alkylthio group include methylthio, ethylthio, propylthio, butylthio, pentylthio, hexylthio group and the like.

In the present specification, the halogen atom refers to fluorine, chlorine, bromine or iodine.

In the present specification, the "C1 to 3 fluoride alkoxy groups" include fluoromethyloxy, difluoromethyloxy, trifluoromethyloxy, fluoroethyloxy, difluoroethyloxy, trifluoroethyloxy, fluoropropyloxy, and difluoropropyloxy. , Trifluoropropyloxy, hexafluoropropyloxy group and the like.

In the present specification, "C1 to 3 alkylenedioxy groups" means methylenedioxy, ethylenedioxy or propylenedioxy groups.

In the present specification, "C3-5 alkylene group" refers to trimethylene, tetramethylene or pentamethylene group.

As used herein, the term "therapeutic or prophylactic agent" means at least one of a "therapeutic agent" and a "preventive agent", and also includes a therapeutic agent and a prophylactic agent.

Hereinafter, preferred embodiments of the compound represented by the general formula (I) will be shown.

As X, CH or N is preferable, and CH is more preferable.

As Y, a carboxyl group or COR ⁴⁰ is preferable, and a carboxyl group is more preferable.

The R ^1, C1-12 alkyl group, preferably a C2~12 alkenyl or C2~12 alkynyl group, more preferably a C1-12 alkyl group, more preferably C2~12 alkyl group, more is C4~10 alkyl group Preferably, a C6-10 alkyl group is particularly preferred.

As R ² represents a hydrogen atom, preferably C1~6 alkyl group and C1~6 alkoxy group, more preferably a hydrogen atom.

Hydrogen atoms as R ^3, preferable C1~6 alkyl group and C1~6 alkoxy, C1 -4 alkyl and C1 -4 alkoxy group is more preferable.

Further, R ² and R ³ may be combined to form a C1 to 3 alkylene dioxy group.

In the compound represented by the general formula (I), X is CH, R ¹ is a C4 to 10 alkyl group, R ² is a hydrogen atom, and R ³ is a C1 to 4 alkyl group or C1 to 4 alkoxy. The base compound is preferred.

A preferred embodiment of the compound represented by the general formula (IA) is shown.

The X ^A, CH or N is preferable, CH is more preferable.

As R ^1A , C1 to 12 alkyl groups are preferable, and C6 to 10 alkyl groups are more preferable.

As R ^2A and R ^3A , a hydrogen atom, a C1-6 alkyl group and a C1-6 alkoxy group are preferable, and a C1-4 alkyl group and a C1-4 alkoxy group are more preferable.

Further, R ^2A and R ^3A may be combined to form a C3 to 5 alkylene group or a C1 to 3 alkylene dioxy group.

Specifically, in the compound represented by the general formula (I) used in the present invention,
Compound 1: 1,4-dihydro-1-methyl-4-oxoquinoline-3-carboxylic acid,
Compound 2: 1,4-dihydro-1-ethyl-4-oxoquinoline-3-carboxylic acid,
Compound 3: 1,4-dihydro-1-propyl-4-oxoquinoline-3-carboxylic acid,
Compound 4: 1,4-dihydro-1-butyl-4-oxoquinoline-3-carboxylic acid,
Compound 5: 1,4-dihydro-1-pentyl-4-oxoquinoline-3-carboxylic acid,
Compound 6: 1,4-dihydro-1-hexyl-4-oxoquinoline-3-carboxylic acid,
Compound 7: 1,4-dihydro-1-heptyl-4-oxoquinoline-3-carboxylic acid,
Compound 8: 1,4-dihydro-1-octyl-4-oxoquinoline-3-carboxylic acid,
Compound 9: 1,4-dihydro-1-cyclopropylmethyl-4-oxoquinoline-3-carboxylic acid,
Compound 10: 1,4-dihydro-1-cyclohexylmethyl-4-oxoquinoline-3-carboxylic acid,
Compound 11: 1,4-dihydro-1-benzyl-4-oxoquinoline-3-carboxylic acid,
Compound 12: 1,4-dihydro-6-methyl-1-propyl-4-oxoquinoline-3-carboxylic acid,
Compound 13: 1,4-dihydro-6-methyl-1-octyl-4-oxoquinoline-3-carboxylic acid,
Compound 14: 1,4-dihydro-7-methyl-1-propyl-4-oxoquinoline-3-carboxylic acid,
Compound 15: 1,4-dihydro-7-methyl-1-octyl-4-oxoquinoline-3-carboxylic acid,
Compound 16: 1,4-dihydro-6-methoxy-1-propyl-4-oxoquinoline-3-carboxylic acid,
Compound 17: 1,4-dihydro-6-methoxy-1-octyl-4-oxoquinoline-3-carboxylic acid,
Compound 18: 1,4-dihydro-7-methoxy-1-propyl-4-oxoquinoline-3-carboxylic acid,
Compound 19: 1,4-dihydro-7-methoxy-1-octyl-4-oxoquinoline-3-carboxylic acid,
Compound 20: 5-ethyl-8-oxo- [1,3] dioxolo [4,5-g] quinoline-7-carboxylic acid (oxophosphate), and compound 29: 5-decyl-8-oxo- [1, 3] Dioxolo [4,5-g] quinoline-7-carboxylic acid is preferable.
Compound 2: 1,4-dihydro-1-ethyl-4-oxoquinoline-3-carboxylic acid,
Compound 4: 1,4-dihydro-1-butyl-4-oxoquinoline-3-carboxylic acid,
Compound 8: 1,4-dihydro-1-octyl-4-oxoquinoline-3-carboxylic acid,
Compound 13: 1,4-dihydro-6-methyl-1-octyl-4-oxoquinoline-3-carboxylic acid,
Compound 15: 1,4-dihydro-7-methyl-1-octyl-4-oxoquinoline-3-carboxylic acid,
Compound 17: 1,4-dihydro-6-methoxy-1-octyl-4-oxoquinoline-3-carboxylic acid,
Compound 19: 1,4-dihydro-7-methoxy-1-octyl-4-oxoquinoline-3-carboxylic acid,
Compound 20: 5-ethyl-8-oxo- [1,3] dioxolo [4,5-g] quinoline-7-carboxylic acid (oxophosphate), and compound 29: 5-decyl-8-oxo- [1, 3] Dioxolo [4,5-g] quinoline-7-carboxylic acid is more preferred.
Compound 13: 1,4-dihydro-6-methyl-1-octyl-4-oxoquinoline-3-carboxylic acid,
Compound 15: 1,4-dihydro-7-methyl-1-octyl-4-oxoquinoline-3-carboxylic acid,
Compound 17: 1,4-dihydro-6-methoxy-1-octyl-4-oxoquinoline-3-carboxylic acid,
Compound 19: 1,4-dihydro-7-methoxy-1-octyl-4-oxoquinoline-3-carboxylic acid,
Compound 20: 5-ethyl-8-oxo- [1,3] dioxolo [4,5-g] quinoline-7-carboxylic acid (oxophosphate) and compound 29: 5-decyl-8-oxo- [1,3 ] Dioxolo [4,5-g] quinoline-7-carboxylic acid is more preferred.

The compounds represented by the general formulas (I) and (IA) include pharmaceutically acceptable salts thereof. Specifically, inorganic bases such as sodium salt, potassium salt, magnesium salt, calcium salt, ammonium salt and aluminum salt, and base addition salts with organic bases such as methylamine, ethylamine, ethanolamine, lysine and ornithine are used. Can be mentioned.

Solvates (including hydrates) and crystalline polymorphs may be present in the compounds represented by the general formulas (I) and (IA) and their pharmaceutically acceptable salts, but alone. It may be in crystalline form or a mixture of multiple crystalline forms, all of which are included. Further, it may be in an amorphous form.

The compounds represented by the general formulas (I) and (IA) or pharmaceutically acceptable salts thereof are known by themselves or substituted by utilizing the characteristics based on the basic skeleton thereof or the type of the substituent thereof. It can be produced by applying various known synthetic methods for group introduction and functional group conversion. For example, 1,4-dihydro-4-oxoquinoline-3-carboxylic acid (DOCA, CAS No .: 13721-01-2) is used as a starting material, and a substituent introduction reaction is carried out by a known method, or Patent Document 2 , Can be produced according to the methods described in Non-Patent Documents 11 and 12. For example, oxolinic acid is a known compound with CAS Number: 14698-29-4.

The compound represented by the general formula (I) can be produced, for example, according to the following reaction step formula 1 or the following examples, but is not limited thereto. Further, the compound represented by the general formula (IA) can also be produced by the same method.

(In the formula, Y ¹ represents a group of Y other than COOH, H represents a halogen atom (iodine, chlorine, bromine), R ^30-1 represents a group of R ³⁰ other than a hydroxyl group, and others. The symbol has the same meaning as above.)

Step (1)
Among the compounds represented by the general formula (I), the compound in which Y is a group other than the COOH group, that is, the compound represented by the general formula (I-1) is a compound represented by the general formula (II). It can be produced by an N-alkylation reaction, which is a reaction with the compound represented by (III). This reaction can be carried out in the presence of an alkaline agent or a strong base, in the presence or absence of a solvent.
The compound represented by the formula (III) is a halogenated hydrocarbon, which is methane chloride, methane bromide, methane iodide, ethane bromide, n-propane chloride, n-propane bromide, isopropane bromide, and bromide. n-butane, t-butane bromide, n-hexane chloride, n-hexane bromide, n-hexane iodide, cyclohexane bromide, n-octane bromide, n-dodecane bromide, n-dodecane iodide, odor Known compounds such as cyclohexyl bromide, benzyl bromide, and allyl bromide can be used.
Examples of the alkaline agent include inorganic alkaline compounds such as sodium hydrogen carbonate, sodium carbonate, potassium carbonate, calcium carbonate, cesium carbonate, sodium hydroxide and potassium hydroxide.
Examples of the strong base include sodium hydride, potassium hydride and the like.
The solvent varies depending on the alkyl halide or alkalizing agent used, but is generally a solvent used for organic synthesis, for example, an alcohol solvent such as methanol, ethanol or 2-propanol, an ether solvent such as diethyl ether, tetrahydrofuran or dioxane. Aromatic solvents such as benzene, toluene and xylene, halogen solvents such as chloroform and dichloromethane, water such as acetonitrile, DMF, DMSO and N-methylpyrrolidone, or mixtures thereof can be used.
Even the general formula Y ¹ group of the compound represented by (II) the compound which was replaced by a COOH group, and the reaction proceeds by N- alkylation and esterification of Step (1) at the same time, the general formula ( The compound represented by I-1) can be produced.

Step (2)
Among the compounds represented by the general formula (I), the compound in which Y is a carboxyl group (the compound represented by the general formula (I-2)) is the general formula (I-1) obtained in step (1). ) Can be obtained by subjecting the compound to a deprotection reaction of an ester group. The deprotection reaction can be carried out according to the method described in Protective Groups in Organic Synthesis, Theodora W Green (John Wiley & Sons) and the like. For example, depending on the protecting group, it is hydrolyzed in the presence of an acid (eg, hydrochloric acid, trifluoroacetic acid) or base (eg, sodium hydroxide, etc.) or catalytically reduced (eg, in the presence of a 10% palladium-carbon catalyst). ) Can be implemented. This reaction can be carried out at room temperature to reflux temperature for 1 to 24 hours.
As the solvent, a solvent generally used for organic synthesis, for example, an alcohol solvent such as methanol, ethanol and 2-propanol, an ether solvent such as diethyl ether, tetrahydrofuran and dioxane, and an aromatic solvent such as benzene, toluene and xylene , Chloroform, dichloromethane and other halogen solvents, acetonitrile, DMF, DMSO, N-methylpyrrolidone and the like, water, or mixtures thereof.

Step (3)
Among the compounds represented by the general formula (I), Y is COOR ⁴⁰ (R ⁴⁰ has the same meaning as described above) and CONR ⁴¹ R ⁴² (R ⁴¹ and R ⁴² have the same meaning as described above). A compound, that is, a compound represented by the general formula (I-3), is produced by subjecting the compound represented by the general formula (I-2) produced in the above step (2) to an esterification or amidation reaction. can do.
The esterification and amidation reactions are, for example,
(A) Method using acid halide,
(B) Method using mixed acid anhydride,
(C) It can be carried out by a method using a condensing agent or the like.

To explain these methods in detail,
In the method using (A) acid halide, for example, the carboxylic acid is mixed with an acid halide agent (oxalyl chloride, thionyl chloride, etc.) in an organic solvent (chloroform, dichloromethane, diethyl ether, tetrahydrofuran, etc.) or in the absence of a solvent. The reaction was carried out at 20 ° C. to reflux temperature, and the obtained acid halide was subjected to alcohol and an organic solvent (chloroform, dichloromethane, diethyl ether, tetrahydrofuran, etc.) in the presence of a base (pyridine, triethylamine, dimethylaniline, dimethylaminopyridine, diisopropylethylamine, etc.) ), It is carried out by reacting at a temperature of about 0 to 40 ° C. It can also be carried out by reacting with acid halide at about 0 to 40 ° C. using an alkaline aqueous solution (sodium bicarbonate solution, sodium hydroxide solution, etc.) in an organic solvent (dioxane, tetrahydrofuran, etc.).
In the method using (B) mixed acid anhydride, for example, the carboxylic acid is used in an organic solvent (chloroform, dichloromethane, diethyl ether, tetrahydrofuran, etc.) or in a solvent-free base (pyridine, triethylamine, dimethylaniline, dimethylaminopyridine, diisopropyl, etc.). It was obtained by reacting with an acid halide (pivaloyl chloride, tosilyl lolide, meshyl lolide, etc.) or an acid derivative (ethyl chloroformate, isobutyl chloroformate, etc.) at about 0 to 40 ° C. in the presence of (ethylamine, etc.). It is carried out by reacting the mixed acid anhydride with an alcohol in an organic solvent (chloroform, dichloromethane, diethyl ether, tetrahydrofuran, etc.) at about 0 to 40 ° C.
(C) In the method using a condensing agent, for example, carboxylic acid and alcohol are mixed in an organic solvent (chloroform, dichloromethane, dimethylformamide, diethyl ether, tetrahydrofuran, etc.) or in a solvent-free base (pyridine, triethylamine, dimethylaniline, etc.). In the presence or absence of (dimethylaminopyridine, etc.)) condensing agents [1,3-dicyclohexylcarbodiimide (DCC), 1-ethyl-3- [3- (dimethylamino) propyl] carbodiimide (EDC), 1,1'. -Carbonyldiimidazole (CDI), 2-chloro-1-methylpyridinium iodine, 1-propylphosphonic acid cyclic anhydride (PPA), etc.] with or without 1-hydroxybenztriazole (HOBt). It is carried out by reacting at 0 to 40 ° C.
It is desirable that the reactions (A), (B) and (C) are all carried out under anhydrous conditions under the atmosphere of an inert gas (for example, argon, nitrogen, etc.).

The compound represented by the general formula (II) used as a starting material is known or a known method, for example, "Comprehensive Organic Transformations: A Guide to Functional Group Preparations 2nd Edition (Richard C. Larock, John Wiley & Sons Inc." , 1999) ”, it can be easily produced. For example, it can be produced by the method represented by the following reaction step formula 2.

(In the formula, R ^p represents an ester protecting group (alkyl group or benzyl group), and other symbols have the same meanings as described above.)

Step (4)
The reaction between the compound represented by the general formula (III) and the compound represented by the general formula (IV) (methylene malonate ester) can be carried out in the absence of a solvent or in the presence of a solvent. As the solvent, water or an organic solvent (for example, ether solvent such as dioxane, tetrahydrofuran, diethyl ether, 1,2-dimethoxyethane, diethylene glycol dimethyl ether, ethylene glycol dimethyl ether; aromatic hydrocarbon solvent such as benzene, toluene, xylene) Lower alcohol solvents such as methanol, ethanol and isopropanol; one or a mixture of polar solvents such as DMF, DMSO, hexamethylphosphate triamide and acetonitrile) are used. The reaction can be carried out at room temperature to 200 ° C., preferably room temperature to 150 ° C. for any time of 1 to 30 hours.

Step (5)
The cyclization reaction using the compound represented by the general formula (V) as a starting compound can be carried out by heating in a solvent such as diphenyl ether as described in Patent Document 1. It can also be carried out by heating the compound represented by the general formula (V) even in the absence of a solvent. Such a reaction is carried out at 150 to 300 ° C. for 5 minutes to 2 hours.

The compounds represented by the general formula (III) and the general formula (IV) are known, or can be produced by a known method using a known compound as a raw material.

The compound represented by the general formula (I) or a pharmaceutically acceptable salt thereof has a ryanodine receptor, particularly ryanodine 1 receptor (RyR1) inhibitory action, and skeletal muscle in animals such as mammals, particularly humans. It can be applied to the treatment of diseases. The compound represented by the general formula (I) or a pharmaceutically acceptable salt thereof is particularly useful for the treatment of malignant hyperthermia and CCD.

When the compound represented by the general formula (I) or a pharmaceutically acceptable salt thereof is used as a medicine, it can be administered orally or parenterally. The compound represented by the general formula (I) or a pharmaceutically acceptable salt thereof can be combined with a pharmaceutically acceptable carrier to form a pharmaceutical composition. Known pharmaceutically acceptable carriers such as excipients, binders, buffers, thickeners, stabilizers, emulsifiers, dispersants, suspending agents and preservatives can be used. , Can be formulated by conventional methods.

Examples of the preparation for oral administration include tablets (including sugar-coated tablets and film-coated tablets), pills, granules, powders, capsules (including soft capsules), syrups, emulsions, suspensions and the like.

This preparation for oral administration can be produced according to a known method by blending additives usually used in the field of preparation. Examples of such additives include excipients such as lactose, mannitol and anhydrous calcium hydrogen phosphate; binders such as hydroxypropyl cellulose, methyl cellulose and polyvinylpyrrolidone; disintegrants such as starch and carboxymethyl cellulose, magnesium stearate, etc. Examples include lubricants such as talc. Parenterally, it can be administered as an injection, a rectal administration preparation, a topical administration, or the like, and an injection is preferable.

Examples of the injection include a sterile solution or suspension. These injections are produced, for example, by dissolving or suspending the compound of the present invention or a pharmaceutically acceptable salt thereof in Japanese Pharmacopoeia water for injection. If necessary, an isotonic agent such as sodium chloride; a buffering agent such as sodium dihydrogen phosphate and sodium monohydrogen phosphate; a solubilizing agent and the like may be blended. In addition, it can be used as a dissolution type (powder-filled, freeze-dried) injection, and in this case, it can be produced by a usual method by adding excipients such as mannitol and lactose.

Examples of the rectal administration preparation include suppositories and the like. A suppository is produced, for example, by dissolving or suspending the compound of the present invention or a pharmaceutically acceptable salt thereof in a base such as cacao fat or macrogol, and then pouring it into a mold to form a suppository. It is also possible to put the liquid or cream in a container for injection to prepare a rectal administration preparation.

Topically administered preparations include liquid preparations, eye drops, creams, ointments, gel preparations, sprays, powders and the like. The liquid preparation can be produced by adding the compound of the present invention or a pharmaceutically acceptable salt thereof to water, and adding a stabilizer, a solubilizer, a thickener, a dispersant, a suspending agent, etc. as necessary. it can. As the thickener, gelatin, sodium hyaluronate, high molecular weight dextran, sodium alginate, sodium chondroitin sulfate and the like can be used. Eye drops can be produced by adding a preservative in addition to a buffering agent, a pH adjuster, and an isotonic agent. Creams and ointments can be produced using aqueous or oily bases such as water, liquid paraffin, vegetable oils (peanut oil, castor oil, etc.), macrogol and the like. As the gel preparation, gelatin, pectin, carrageenan, agar, tragant, alginate, cellulose ether (methyl cellulose, sodium carboxymethyl cellulose, etc.), pectin derivative, polyacrylate, polymethacrylate, polyvinyl alcohol, polyvinylpyrrolidone, etc. are used by a known method. Can be manufactured. The spray agent can be produced by dissolving or suspending the compound of the present invention or a pharmaceutically acceptable salt thereof in water or the like and then putting it in a spray container. In the case of a powder, the compound of the present invention or a pharmaceutically acceptable salt thereof can be used as it is, but it can be produced by mixing with an appropriate excipient.

The dose of the compound of the present invention is appropriately determined according to each individual case in consideration of the target disease or symptom, the age, body weight, sex, etc. of the subject to be administered. Usually, in the case of oral administration, the daily dose of the compound of the present invention for an adult (body weight of about 60 kg) is 0.01 to 100 mg, preferably 0.01 to 30 mg, and more preferably 0.1 to 10 mg. Is administered once or in 2 to 4 divided doses. When administered intravenously, the daily dose for an adult is usually 0.03 to 3000 μg, preferably 0.03 to 300 μg, more preferably 0.03 to 30 μg per 1 kg of body weight, and 1 day. Administer in divided doses to multiple doses.

[Example]
Hereinafter, the present invention will be specifically described with reference to examples, but the present invention is not limited thereto.

[Synthesis of test compound]
Reference example 1
Methyl 1,4-dihydro-1-methyl-4-oxoquinoline-3-carboxylate

In a 30 mL eggplant-shaped flask, 1,4-dihydro-4-oxoquinoline-3-carboxylic acid (200 mg, 1.06 mmol) and methyl iodide (450 mg, 3.18 mmol) were added to N, N-dimethylformamide (3 mL). ), Further cesium carbonate (1.03 g) was added, and the mixture was stirred at 50 ° C. After 6 hours, chloroform and water were added to the reaction solution to separate the solutions. The organic layer was dried over anhydrous sodium sulfate and concentrated under reduced pressure to give the title compound as a colorless liquid (quantitative).
¹ H-NMR (500 MHz, CDCl ₃ ): δ 8.56 (dd, J = 1.5, 8.1 Hz, 1 H), 8.53 (s, 1 H), 7.73 (ddd, J = 8.5, 7.0, 1.6 Hz, 1) H), 7.49 (ddd, J = 7.2, 8.0, 0.9 Hz, 1 H), 7.45 (d, J = 8.5 Hz, 1 H), 3.95 (s, 3 H), 3.91 (s, 3 H).

Synthesis example 1
1,4-Dihydro-1-methyl-4-oxoquinoline-3-carboxylic acid (Compound 1)

The compound (0.37 mmol) prepared in Reference Example 1 was dissolved in methanol (3 mL) in a 30 mL eggplant-shaped flask. A 1 M aqueous sodium hydroxide solution (2 mL) was added dropwise thereto, and the mixture was stirred at room temperature. After 3 hours, 1 M hydrochloric acid was added to the reaction solution, and the precipitated white solid was recovered by suction filtration. The obtained solid was recrystallized from a mixed solvent of n-hexane and ethyl acetate to give the title compound as colorless needle-like crystals (yield: 96%).
¹ H-NMR (500 MHz, CDCl ₃ ): δ8.56 (dd, J = 8.1, 1.5 Hz, 1 H), 8.53 (s, 1 H), 7.73 (ddd, J = 8.5, 7.0, 1.5 Hz, 1 H), 7.49 (ddd, J = 8.0, 7.2, 0.9 Hz, 1 H), 7.45 (d, J = 8.5 Hz, 1 H), 3.95 (s, 3 H), 3.91 (s, 3 H).

Synthesis Example 1 (1) to Synthesis Example 1 (11)
Using the corresponding alkyl iodide compound instead of methyl iodide, the following compound was produced by the same method as that shown in Reference Example 1 → Synthesis Example 1.

Synthesis Example 1 (1)
1,4-Dihydro-1-ethyl-4-oxoquinoline-3-carboxylic acid (Compound 2)

¹ H-NMR (500 MHz, CDCl ₃ ): δ 15.0 (s, 1 H), 8.76 (s, 1 H), 8.54 (dd, J = 8.2, 1.5 Hz, 1 H), 7.85 (ddd, J = 8.8, 7.2, 1.6 Hz, 1 H), 7.60 (d, J = 8.8 Hz, 1 H), 7.62 (ddd, J = 8.1, 7.2, 0.9 Hz, 1 H), 4.41 (q, J = 7.5 Hz, 2 H), 1.58 (t, J = 7.5 Hz, 3 H).
Synthesis example 1 (2)
1,4-Dihydro-1-propyl-4-oxoquinoline-3-carboxylic acid (Compound 3)

¹ H-NMR (500 MHz, CDCl ₃ ): δ15.0 (s, 1 H), 8.77 (s, 1 H), 8.58 (dd, J = 8.2, 1.5 Hz, 1 H), 7.86 (ddd, J) = 8.7, 7.1, 1.6 Hz, 1 H), 7.63 (d, J = 8.7 Hz, 1 H), 7.60 (ddd, J = 8.1, 7.2, 0.9 Hz, 1 H), 4.31 (t, J = 7.4 Hz , 2 H), 2.00 (sex, J = 7.4 Hz, 2 H), 1.07 (t, J = 7.4 Hz, 3 H).
Synthesis example 1 (3)
1,4-Dihydro-1-butyl-4-oxoquinoline-3-carboxylic acid (Compound 4)

¹ H-NMR (500 MHz, CDCl ₃ ): δ14.96 (s, 1 H), 8.76 (s, 1 H), 8.56 (dd, J = 8.1, 1.5 Hz, 1 H), 7.85 (ddd, J) = 8.7, 7.1, 1.6 Hz, 1 H), 7.63 (d, J = 8.6 Hz, 1 H), 7.59 (ddd, J = 8.1, 7.3, 0.2 Hz, 1 H), 4.33 (m, 2 H), 1.92 (t, J = 7.6 Hz, 2 H), 1.46 (sex, J = 7.56 Hz, 2 H), 1.01 (t, J = 7.4 Hz, 3 H).
Synthesis Example 1 (4)
1,4-Dihydro-1-pentyl-4-oxoquinoline-3-carboxylic acid (Compound 5)

¹ H-NMR (500 MHz, CDCl ₃ ): δ 14.97 (s, 1 H), 8.76 (s, 1 H), 8.57 (dd, J = 8.1, 1.5, Hz, 1 H), 7.85 (ddd, J) = 8.7, 7.1, 1.6 Hz, 1 H), 7.62 (d, J = 8.5 Hz, 1 H), 7.60 (ddd, J = 8.1, 7.2, 0.7 Hz, 2 H), 4.31 (t, J = 7.6 Hz , 2 H), 1.94 (quin, J = 7.5 Hz, 2 H), 1.40 (m, 4 H), 0.93 (t, J = 7.1 Hz, 3 H).
Synthesis Example 1 (5)
1,4-Dihydro-1-hexyl-4-oxoquinoline-3-carboxylic acid (Compound 6)

¹ H-NMR (500 MHz, CDCl ₃ ): δ14.95 (s, 1 H), 8.76 (s, 1 H), 8.58 (dd, J = 8.1, 1.4 Hz, 1 H), 7.85 (ddd, J) = 8.7, 7.1, 1.6 Hz, 1 H), 7.62 (d, J = 8.7 Hz, 2 H), 7.60 (td, J = 7.8, 0.6, Hz, 2 H), 4.31 (t, J = 7.6 Hz, 2 H), 1.94 (quin, J = 7.5 Hz, 2 H), 1.43 (m, 2 H), 1.34 (m, 4 H), 0.90 (t, J = 7.1 Hz, 3 H).
Synthesis Example 1 (6)
1,4-Dihydro-1-heptyl-4-oxoquinoline-3-carboxylic acid (Compound 7)

¹ H-NMR (500 MHz, CDCl ₃ ): δ14.96 (s, 1 H) 8.76 (s, 1 H), 8.57 (dd, J = 8.1, 1.4 Hz, 1 H), 7.85 (ddd, J = 8.7, 7.1, 1.6 Hz, 1 H), 7.62 (d, J = 8.7 Hz, 2 H), 7.60 (ddd, J = 7.9, 7.2, 0.7 Hz, 2 H), 4.31 (t, J = 7.5 Hz, 2 H), 1.94 (quin, J = 7.5 Hz, 2 H), 1.39 (m, 2 H), 1.29 (m, 6 H), 0.88 (t, J = 6.9 Hz, 3 H).
Synthesis Example 1 (7)
1,4-Dihydro-1-octyl-4-oxoquinoline-3-carboxylic acid (Compound 8)

¹ H-NMR (500 MHz, CDCl ₃ ): δ 14.97 (s, 1 H), 8.76 (s, 1 H), 8.58 (dd, J = 8.1, 1.5 Hz, 1 H), 7.85 (ddd, J = 8.7, 7.1, 1.6 Hz, 1 H), 7.62 (d, J = 8.6 Hz, 1 H), 7.60 (t, J = 7.6 Hz, 1 H), 4.31 (t, J = 7.6 Hz, 2 H), 1.94 (quin, J = 7.6 Hz, 2 H), 1.23-1.45 (m, 10 H), 0.88 (t, J = 7.5 Hz, 3 H).
Synthesis Example 1 (8)
1,4-Dihydro-1-cyclopropylmethyl-4-oxoquinoline-3-carboxylic acid (Compound 9)

¹ H-NMR (500 MHz, CDCl ₃ ): δ14.99 (s, 1 H), 8.85 (s, 1 H), 8.58 (dd, J = 8.1, 1.5 Hz, 1 H), 7.86 (ddd, J) = 8.6, 7.1, 1.6 Hz, 1 H), 7.74 (d, J = 8.6 Hz, 1 H), 7.61 (ddd, J = 8.0, 7.2, 0.8 Hz, 1 H), 4.18 (d, J = 7.1 Hz) , 2 H), 1.41 (m, 1 H), 0.81 (m, 2 H), 0.52 (m, 2 H).
Synthesis Example 1 (9)
1,4-Dihydro-1-cyclohexylmethyl-4-oxoquinoline-3-carboxylic acid (Compound 10)

¹ H-NMR (500 MHz, CDCl ₃ ): δ 14.96 (s, 1 H), 8.68 (s, 1 H), 8.58 (dd, J = 8.4, 1.6 Hz, 1 H), 7.85 (ddd, J = 8.6, 7.1, 1.6 Hz, 1 H), 7.62-7.59 (m, 2 H), 4.13 (d, J = 7.4 Hz, 2 H), 1.94 (m, 1 H), 1.80-1.60 (m, 5 H) ), ・ 1.25-1.06 (m, 5 H).
Synthesis Example 1 (10)
1,4-Dihydro-1-benzyl-4-oxoquinoline-3-carboxylic acid (Compound 11)

¹ H-NMR (500 MHz, CDCl ₃ ): δ14.89 (s, 1 H), 8.91 (s, 1 H), 8.57 (dd, J = 8.1, 1.5 Hz, 1 H), 7.72 (ddd, J) = 8.6, 7.1, 1.6 Hz, 1 H), 7.55 (1 H, ddd, J = 8.3, 7.4, 0.7 Hz), 7.53 (d, J = 8.61 Hz, 1 H), 7.40-7.35 (m, 5 H), 7.17 (m, 2 H), 5.53 (s, 2 H).
Synthesis Example 1 (11)
1-Cyclohexylmethyl-1,4-dihydro-4-oxoquinoline-3-carboxylic acid (Compound 35)

^{1 1} H-NMR (500 MHz, CDCl ₃ ) δ 14.96 (s, 1 H), 8.68 (s, 1 H), 8.58 (dd, J = 8.4, 1.6 Hz, 1 H), 7.85 (ddd, J = 8.6) , 7.1, 1.6 Hz, 1 H), 7.62-7.59 (m, 2 H), 4.13 (d, J = 7.4 Hz, 2 H), 1.94 (m, 1 H), 1.80-1.60 (m, 5 H) , 1.25-1.06 (m, 5 H).
Reference example 2
Diethyl p-toluidinomethylene malonate

In a 30 mL eggplant-shaped flask, p-toluidine (1.74 g, 16.2 mmol) was dissolved in ethanol (20 mL). Diethyl ethoxymethylene malonate (4.0 g, 16.3 mmol) was added dropwise while stirring the mixture, and the mixture was stirred at room temperature for 18 hours. After distilling off ethanol, the residue was separated by ethyl acetate and water, the organic layer was dried over anhydrous sodium sulfate, concentrated under reduced pressure, and the residue was subjected to flash column chromatography (n-hexane: ethyl acetate = 4: 1). Purification gave the title compound as a colorless liquid (yield: 90%).
¹ H-NMR (500 MHz, CDCl ₃ ): δ 10.98 (d, J = 13.4 Hz, 1 H), 8.50 (d, J = 13.8 Hz, 1 H), 7.17 (d, J = 8.8 Hz, 1 H) ), 7.03 (d, J = 8.5 Hz, 1 H), 4.30 (q, J = 7.4 Hz, 2 H), 4.24 (q, J = 7.1 Hz, 2 H), 1.37 (t, J = 7.1 Hz, 3 H), 1.32 (t, J = 7.12 Hz, 3 H).
Reference example 3
Ethyl 1,4-dihydro-6-methyl-4-oxoquinoline-3-carboxylate

After heating diphenyl ether (50 mL) to 220 ° C. in a 200 mL two-necked flask, a diphenyl ether (10 ml) solution of the compound (6.4 mmol) produced in Reference Example 2 was added dropwise over 30 minutes at 220 ° C. for 10 hours. Stirred. After returning the reaction solution to room temperature, diethyl ether and n-hexane were added to precipitate a solid. The precipitated solid was collected by suction filtration and washed with diethyl ether to give compound (C) as a milky white solid (yield: 40%).
¹ H-NMR (500 MHz, CDCl ₃ ): δ 12.24 (s, 1 H), 8.50 (s, 1 H), 7.94 (s, 1 H), 7.52 (s, 2 H), 4.21 (q, J) = 7.1 Hz, 2 H), 2.42 (s, 3 H), 1.28 (t, J = 7.1 Hz, 3 H).
Reference example 4
Ethyl 1,4-dihydro-6-methyl-1-propyl-4-oxoquinoline-3-carboxylate

In a 30 mL two-necked flask, 5 mL of dried N, N-dimethylformamide was added to sodium hydride (0.86 mmol) washed with n-hexane. Under ice-cooling, a solution of the compound (0.41 mmol) prepared in Reference Example 3 in N, N-dimethylformamide (1 mL) was added dropwise, and propyl iodide (0.86 mmol) was further added dropwise. After stirring at 55 ° C. for 9 hours, the temperature was returned to room temperature, and a saturated aqueous solution of ammonium chloride was added to stop the reaction. Ethyl acetate and water were added to separate the liquids, and the organic layer was dried over anhydrous sodium sulfate and concentrated under reduced pressure. The residue was purified by open column chromatography (n-hexane: ethyl acetate = 4: 1) to give the title compound as a colorless liquid (yield: 72%).
¹ H-NMR (500 MHz, CDCl ₃ ): δ8.45 (s, 1 H), 8.36 (d, J = 1.0 Hz, 1 H), 7.50 (dd, J = 8.7, 2.1 Hz, 1 H), 7.35 (d, J = 8.6 Hz, 1 H), 4.30 (t, J = 6.8 Hz, 2 H), 4.14 (t, J = 7.3 Hz, 2 H), 2.48 (s, 3 H), 1.94 (sex) , J = 7.4 Hz, 3 H), 1.83 (sex, J = 7.2 Hz, 3 H), 1.06 (t, J = 7.2 Hz, 3 H), 1.03 (t, J = 7.4 Hz, 3 H).
Synthesis example 2
1,4-Dihydro-6-methyl-1-propyl-4-oxoquinoline-3-carboxylic acid (Compound 12)

In a 30 mL eggplant-shaped flask, the compound (0.32 mmol) prepared in Compound Reference Example 4 was dissolved in methanol (3 mL). A 1 M aqueous sodium hydroxide solution (2 mL) was added dropwise thereto, and the mixture was stirred at room temperature. After 3 hours, 1 M hydrochloric acid solution was added to the reaction solution, and the precipitated white solid was collected by suction filtration. The obtained solid was recrystallized from a mixed solvent of n-hexane and ethyl acetate to give the title compound as colorless needle-like crystals (yield: 74%).
¹ H-NMR (500 MHz, CDCl ₃ ): δ15.09 (s, 1 H) 8.73 (s, 1 H), 8.37 (d, J = 0.9 Hz, 1 H), 7.66 (dd, J = 8.8, 2.1 Hz, 1 H), 7.53 (d, J = 8.8 Hz, 1 H), 4.28 (t, J = 7.4 Hz, 2 H), 2.55 (s, 3 H), 1.99 (sex, J = 7.4 Hz, 2 H), 1.05 (t, J = 7.4 Hz, 3 H).

Synthesis Example 2 (1) -Synthesis Example 2 (28)
Reference Example 2 (using a corresponding compound in place of p-toluidine if necessary) → Reference Example 3 → Reference Example 4 (using an equivalent alkyl iodide in place of ethyl iodide if necessary) → The following compounds were produced by the same method as that shown in Synthesis Example 2.

Synthesis example 2 (1)
1,4-Dihydro-6-methyl-1-octyl-4-oxoquinoline-3-carboxylic acid (Compound 13)

¹ H-NMR (500 MHz, CDCl ₃ ): δ15.10 (s, 1 H), 8.73 (s, 1 H), 8.37 (d, J = 0.9 Hz, 1 H), 7.67 (q, J = 3.6) Hz, 1 H), 7.52 (d, J = 8.8 Hz, 1 H), 4.30 (t, J = 7.5 Hz, 2 H), 2.55 (s, 3 H), 1.93 (quin, J = 7.5 Hz, 2 H), 1.44-1.26 (m, 10 H), 0.89 (t, J = 7.0 Hz, 3 H).
Synthesis example 2 (2)
1,4-Dihydro-7-methyl-1-propyl-4-oxoquinoline-3-carboxylic acid (Compound 14)

¹ H-NMR (500 MHz, CDCl ₃ ): δ 15.08 (s, 1 H), 8.72 (s, 1 H), 8.45 (d, J = 8.3 Hz, 1 H), 7.42 (br d, J = 8.3) Hz, 1 H), 7.37 (br s, 1 H), 4.27 (t, J = 7.4 Hz, 2 H), 2.61 (s, 3 H), 1.99 (sex, J = 7.4 Hz, 2 H), 1.07 (t, J = 7.4 Hz, 3 H).
Synthesis example 2 (3)
1,4-Dihydro-7-methyl-1-octyl-4-oxoquinoline-3-carboxylic acid (Compound 15)

¹ H-NMR (500 MHz, CDCl ₃ ): δ15.09 (s, 1 H), 8.72 (s, 1 H), 8.45 (d, J = 8.3 Hz, 1 H), 7.42 (br d, J = 8.3 Hz, 1 H), 7.37 (br s, 1 H), 4.29 (t, J = 7.5 Hz, 2 H), 2.61 (s, 3 H), 1.93 (quin, J = 7.5 Hz, 2 H), 1.45-1.23 (m, 10 H), 0.89 (t, J = 6.9 Hz, 3 H).
Synthesis example 2 (4)
1,4-Dihydro-6-methoxy-1-propyl-4-oxoquinoline-3-carboxylic acid (Compound 16)

¹ H-NMR (500 MHz, DMSO-d ₆ ): δ 15.42 (s, 1 H), 8.98 (s, 1 H), 8.03 (d, J = 9.4 Hz, 1 H), 7.75 (d, J = 3.0 Hz, 1 H), 7.57 (dd, J = 9.4, 3.0 Hz, 1 H), 4.53 (t, J = 7.30 Hz, 2 H), 3.92 (s, 3 H), 1.81 (sex, J = 7.4 Hz, 2 H), ... 0.90 (t, J = 7.4 Hz, 3 H).
Synthesis Example 2 (5)
1,4-Dihydro-6-methoxy-1-octyl-4-oxoquinoline-3-carboxylic acid (Compound 17)

¹ H-NMR (500 MHz, DMSO-d ₆ ): δ 15.42 (s, 1 H), 8.97 (s, 1 H), 8.02 (d, J = 9.5 Hz, 1 H), 7.75 (d, J = 3.0 Hz, 1 H), 7.58 (dd, J = 3.1, 9.4 Hz, 1 H), 4.56 (t, J = 7.3 Hz, 2 H), 3.92 (s, 3 H), 1.77 (quin, J = 7.2 Hz, 2 H), 1.34-1.18 (m, 10 H), 0.83 (t, J = 6.90 Hz, 3 H).
Synthesis Example 2 (6)
1,4-Dihydro-7-methoxy-1-propyl-4-oxoquinoline-3-carboxylic acid (Compound 18)

¹ H-NMR (500 MHz, CDCl ₃ ): δ15.15 (s, 1 H), 8.70 (s, 1 H), 8.50 (d, J = 9.1 Hz, 1 H), 7.18 (dd, J = 9.1) , 2.0 Hz, 1 H), 6.93 (d, J = 1.9 Hz, 1 H), 4.22 (t, J = 7.3 Hz, 2 H), 3.99 (s, 3 H), 1.99 (sex, J = 7.3 Hz , 2 H), 1.06 (t, J = 7.4 Hz, 3 H).
Synthesis Example 2 (7)
1,4-Dihydro-7-methoxy-1-octyl-4-oxoquinoline-3-carboxylic acid (Compound 19)

¹ H-NMR (500 MHz, CDCl ₃ ): δ 15.16 (s, 1 H), 8.69 (s, 1 H), 8.50 (d, J = 9.1 Hz, 1 H), 7.18 (dd, J = 9.1, 2.1 Hz, 1 H), 6.93 (d, J = 1.9 Hz, 1 H), 4.24 (t, J = 7.48 Hz, 2 H), 3.99 (s, 3 H), 1.93 (quin, J = 7.4 Hz, 2 H), 0.89 (t, J = 7.0 Hz, 3 H).
Synthesis Example 2 (8)
1,4-Dihydro-6-methyl-1-octyl-4-oxoquinoline-3-carboxylic acid (Compound 21)

¹ H-NMR (500 MHz, CDCl ₃ ) δ 15.10 (s, 1 H), 8.73 (s, 1 H), 8.37 (d, J = 0.9 Hz, 1 H), 7.67 (q, J = 3.6 Hz, 1 H), 7.52 (d, J = 8.8 Hz, 1 H), 4.30 (t, J = 7.5 Hz, 2 H), 2.55 (s, 3 H), 1.93 (quin, J = 7.5 Hz, 2 H) , 1.44-1.26 (m, 10 H), 0.89 (t, J = 7.0 Hz, 3 H).
Synthesis Example 2 (9)
1,4-Dihydro-7-methyl-1-propyl-4-oxoquinoline-3-carboxylic acid (Compound 22)

^{1 1} H-NMR (500 MHz, CDCl ₃ ) δ 15.08 (s, 1 H), 8.72 (s, 1 H), 8.45 (d, J = 8.3 Hz, 1 H), 7.42 (br d, J = 8.3 Hz , 1 H), 7.37 (br s, 1 H), 4.27 (t, J = 7.4 Hz, 2 H), 2.61 (s, 3 H), 1.99 (sex, J = 7.4 Hz, 2 H), 1.07 ( t, J = 7.4 Hz, 3 H).
Synthesis Example 2 (10)
1,4-Dihydro-7-methyl-1-octyl-4-oxoquinoline-3-carboxylic acid (Compound 23)

^{1 1} H-NMR (500 MHz, CDCl ₃ ) δ 15.09 (s, 1 H), 8.72 (s, 1 H), 8.45 (d, J = 8.3 Hz, 1 H), 7.42 (br d, J = 8.3 Hz , 1 H), 7.37 (br s, 1 H), 4.29 (t, J = 7.5 Hz, 2 H), 2.61 (s, 3 H), 1.93 (quin, J = 7.5 Hz, 2 H), 1.45- 1.23 (m, 10 H), 0.89 (t, J = 6.9 Hz, 3 H).
Synthesis Example 2 (11)
1,4-Dihydro-6-methoxy-1-octyl-4-oxoquinoline-3-carboxylic acid (Compound 24)

¹ H-NMR (500 MHz, DMSO-d ₆ ) δ 15.42 (s, 1 H), 8.97 (s, 1 H), 8.02 (d, J = 9.5 Hz, 1 H), 7.75 (d, J = 3.0) Hz, 1 H), 7.58 (dd, J = 3.1, 9.4 Hz, 1 H), 4.56 (t, J = 7.3 Hz, 2 H), 3.92 (s, 3 H), 1.77 (quin, J = 7.2 Hz , 2 H), 1.34-1.18 (m, 10 H), 0.83 (t, J = 6.90 Hz, 3 H).
Synthesis Example 2 (12)
1,4-Dihydro-7-methoxy-4-oxo-1-propylquinoline-3-carboxylic acid (Compound 25)

¹ H-NMR (500 MHz, CDCl ₃ ) δ 15.15 (s, 1 H), 8.70 (s, 1 H), 8.50 (d, J = 9.1 Hz, 1 H), 7.18 (dd, J = 9.1, 2.0 Hz, 1 H), 6.93 (d, J = 1.9 Hz, 1 H), 4.22 (t, J = 7.3 Hz, 2 H), 3.99 (s, 3 H), 1.99 (sext, J = 7.3 Hz, 2 H), 1.06 (t, J = 7.4 Hz, 3 H)
Synthesis Example 2 (13)
1,4-Dihydro-7-methoxy-1-octyl-4-oxoquinoline-3-carboxylic acid (Compound 26)

^{1 1} H-NMR (500 MHz, CDCl ₃ ) δ 15.16 (s, 1 H), 8.69 (s, 1 H), 8.50 (d, J = 9.1 Hz, 1 H), 7.18 (dd, J = 9.1, 2.1) Hz, 1 H), 6.93 (d, J = 1.9 Hz, 1 H), 4.24 (t, J = 7.48 Hz, 2 H), 3.99 (s, 3 H), 1.93 (quin, J = 7.4 Hz, 2 H), 0.89 (t, J = 7.0 Hz, 3 H).
Synthesis Example 2 (14)
1,4-Dihydro-1-propyl-4-oxo-7-trifluoromethylquinoline-3-carboxylic acid (Compound 27)

¹ H-NMR (400 MHz, CDCl ₃ ) δ 14.5 (s, 1H), 8.83 (s, 1 H), 8.71 (d, J = 8.4 Hz, 1 H), 7.85 (s, 1 H), 7.81 ( d, J = 8.8 Hz, 1 H), 4.34 (t, J = 7.4 Hz, 2 H), 2.01 (sext, J = 7.4 Hz, 2 H), 0.83 (t, J = 7.4 Hz, 3 H).
Synthesis Example 2 (15)
1,4-Dihydro-1-octyl-4-oxo-7-trifluoromethylquinoline-3-carboxylic acid (Compound 28)

¹ H-NMR (400 MHz, CDCl ₃ ) δ 14.5 (s, 1H), 8.82 (s, 1 H), 8.71 (d, J = 8.4 Hz, 1 H), 7.85 (s, 1 H), 7.80 ( d, J = 8.5 Hz, 1 H), 4.35 (t, J = 7.5 Hz, 2H), 1.95 (quin, J = 7.3 Hz, 2 H), 1.28-1.46 (m, 10 H), 0.88 (t, J = 6.8 Hz, 3 H).
Synthesis Example 2 (16)
5-decyl-8-oxo- [1,3] dioxolo [4,5-g] quinoline-7-carboxylic acid (Compound 29)

^{1 1} H-NMR (500 MHz, CDCl ₃ ) 9.21 (s, 1 H), 7.60 (s, 1 H), 7.49 (s, 1 H), 6.18 (s, 2 H), 4.42 (t, J = 6.5) Hz, 2 H), 1.91 (quin, J = 6.5 Hz, 2 H), 1.48 (q, J = 7.5 Hz, 2 H), 1.37-1.25 (m, 12 H), 0.87 (t, J = 5.2 Hz) , 3 H).
Synthesis Example 2 (17)
7-Methoxy-1-propyl-1,4-dihydro-4-oxo-1,8-naphthalene-3-carboxylic acid (Compound 30)

^{1 1} H-NMR (400 MHz, CDCl ₃ ) δ 15.17 (s, 1 H), 9.09 (s, 1 H), 8.50 (d, J = 8.8 Hz, 1 H), 7.11 (d, J = 8.8, 1 H), 4.53 (t, J = 6.8 Hz, 2 H), 4.05 (s, 3 H), 1.88 (sext, J = 7.2 Hz, 2 H), 0.91 (t, J = 7.2 Hz, 3 H).
Synthesis Example 2 (18)
7-Methoxy-1-octyl-1,4-dihydro-4-oxo-1,8-naphthylidine-3-carboxylic acid (Compound 31)

^{1 1} H-NMR (400 MHz, CDCl ₃ ) δ 15.17 (s, 1 H), 9.08 (s, 1 H), 8.55 (d, J = 8.8 Hz, 1 H), 7.11 (d, J = 8.8, 1 H), 4.55 (t, J = 7.2 Hz, 2 H), 4.05 (s, 3 H), 1.84 (quin, J = 6.4 Hz, 2 H), 1.39-1.18 (m, 10 H), 0.83 (t) , J = 7.2 Hz, 3 H).
Synthesis Example 2 (19)
1-decyl-7-methoxy-1,4-dihydro-4-oxo-1,8-naphthalene-3-carboxylic acid (Compound 32)

^{1 1} H-NMR (500 MHz, CDCl ₃ ) δ 15.17 (s, 1 H), 9.08 (s, 1 H), 8.55 (d, J = 8.5 Hz, 1 H), 7.11 (d, J = 8.5, 1 H), 4.55 (t, J = 7.5 Hz, 2 H), 4.05 (s, 3 H), 1.84 (quin, J = 7.0 Hz, 2 H), 1.38-1.15 (m, 14 H), 0.83 (t) , J = 6.5 Hz, 3 H).
Synthesis Example 2 (20)
1,4-Dihydro-6,7-dimethoxy-1-octyl-4-oxoquinoline-3-carboxylic acid (Compound 33)

¹ H-NMR (400 MHz, CDCl ₃ ) 9.23 (s, 1 H), 8.01 (s, 1 H), 7.52 (s, 1 H), 4.60 (t, J = 6.4 Hz, 2 H), 4.10 ( s, 3 H), 4.02 (s, 3H), 1.89 (quin, J = 6.4 Hz, 2 H), 1.49 (quin, J = 6.8 Hz, 2 H), 1.35-1.20 (m, 8 H), 0.84 (t, J = 6.8 Hz, 3 H).
Synthesis Example 2 (21)
1-decyl-1,4-dihydro-6,7-dimethoxy-4-oxoquinoline-3-carboxylic acid (Compound 34)

¹ H-NMR (400 MHz, CDCl ₃ ) 8.66 (s, 1 H), 7.86 (s, 1 H), 6.88 (s, 1 H), 4.27 (t, J = 14.8 Hz, 2 H), 4.05 ( s, 3 H), 4.04 (s, 3H), 1.94 (quin, J = 7.4 Hz, 2 H), 1.43-1.22 (m, 14 H), 0.87 (t, J = 13.6 Hz, 3 H).

Synthesis Example 2 (22)
4-Oxo-1-propyl-cyclopenta [g] quinoline-3-carboxylic acid (Compound 35)

¹ H-NMR (500 MHz, CDCl ₃ ) δ 15.28 (br s, 1H), 8.68 (s, 1 H), 8.37 (s, 1 H), 7.43 (s, 1 H), 4.26 (t, J = 7.0 Hz, 2 H), 3.12 (t, J = 7.5 Hz, 2 H), 3.06 (t, J = 7.5 Hz, 2 H), 2.21 (quin, J = 7.0 Hz, 2 H), 1.92 (sext, J = 7.5 Hz, 2 H), 1.04 (t, J = 7.5 Hz, 3 H).
Synthesis Example 2 (23)
1-octyl-4-oxo-cyclopenta [g] quinoline-3-carboxylic acid (Compound 36)

¹ H-NMR (500 MHz, CDCl ₃ ) δ 15.3 (s, 1H), 8.68 (s, 1 H), 8.37 (s, 1 H), 7.42 (s, 1 H), 4.27 (t, J = 7.5) Hz, 2 H), 3.12 (t, J = 7.4 Hz, 2H), 3.08 (t, J = 7.6 Hz, 2H), 2.21 (quin, J = 7.5 Hz, 2 H), 1.92 (quin, J = 7.4 Hz, 2 H), 1.25-1.41 (m, 10 H), 0.88 (t, J = 6.8 Hz, 3 H).
Synthesis Example 2 (24)
1,4-Dihydro-1-propyl-7-trifluoromethyloxy-4-oxoquinoline-3-carboxylic acid (Compound 37)

^{1 1} H-NMR (400 MHz, CDCl ₃ ) δ 14.59 (s, 1H), 8.78 (s, 1 H), 8.62 (d, J = 8.8 Hz, 1 H), 7.44 (d, J = 8.8 Hz, 1 H), 7.39 (s, 1 H), 4.26 (t, J = 7.6 Hz, 2H), 1.99 (sext, J = 7.2 Hz, 2 H), 1.07 (t, J = 7.2 Hz, 3 H).
Synthesis Example 2 (25)
1,4-Dihydro-1-propyl-7-trifluoromethyloxy-4-oxoquinoline-3-carboxylic acid (Compound 38)

¹ H-NMR (400 MHz, CDCl ₃ ) δ 14.6 (s, 1H), 8.76 (s, 1 H), 8.62 (d, J = 9.0 Hz, 1 H), 7.43 (d, J = 9.0 Hz, 1 H), 7.38 (s, 1 H), 4.26 (t, J = 7.5 Hz, 2H), 1.92 (quin, J = 7.2 Hz, 2 H), 1.28-1.44 (m, 10 H), 0.88 (t, J = 7.9 Hz, 3 H).
Synthesis Example 2 (26)
1,4-Dihydro-8-methoxy-1-octyl-4-oxoquinoline-3-carboxylic acid (Compound 39)

¹ H-NMR (400 MHz, CDCl ₃ ) 8.62 (s, 1 H), 8.18 (dd, J = 8.0, 1.2 Hz, 1 H), 7.50 (t, J = 8.0 Hz, 1 H), 7.30 (dd) , J = 8.8, 1.2 Hz, 1 H), 4.60 (t, J = 7.6 Hz, 2 H), 4.02 (s, 3 H), 1.84 (quin, J = 7.2 Hz, 2 H), 1.32-1.26 ( m, 10 H), 0.88 (t, J = 6.8 Hz, 3 H). Synthesis Example 2 (27)
7,8-Dimethoxy-1,4-dihydro-1-octyl-4-oxoquinoline-3-carboxylic acid (Compound 40)

¹ H-NMR (400 MHz, CDCl ₃ ) 8.61 (s, 1 H), 8.34 (d, J = 8.8 Hz, 1 H), 7.24 (d, J = 9.2 Hz, 1 H), 4.49 (t, J) = 7.6 Hz, 2 H), 4.06 (s, 3 H), 3.93 (s, 3 H), 1.83 (quin, J = 7.2 Hz, 2 H), 1.30-1.25 (m, 10 H), 0.87 (t , J = 6.8 Hz, 3H).
Synthesis Example 2 (28)
1,4-Dihydro-7-methylthio-1-octyl-4-oxoquinoline-3-carboxylic acid (Compound 41)

^{1 1} H-NMR (400 MHz, CDCl ₃ ) 8.68 (s, 1 H), 8.42 (d, J = 8.8 Hz, 1 H), 7.39 (dd, J = 8.8, 1.6 Hz, 1 H), 7.28 (d) , J = 1.2 Hz, 1 H), 4.26 (t, J = 7.2 Hz, 2 H), 2.62 (s, 3 H), 1.92 (quin, J = 7.6 Hz, 2 H), 1.45-1.22 (m, 10 H), 0.88 (t, J = 6.8 Hz, 3 H).

Pharmacological test 1 Inhibitory activity test against RyR1 The compound of the present invention was tested for its inhibitory activity against RyR1 by the following method. The test method is based on Ca ²⁺ concentration ([Ca ²⁺ ] _ER ) in the vesicles of HEK293 cells expressing a disease variant (malignant hyperthermia, central core disease) with reference to Non-Patent Document 10. The degree of increase in Ca ²⁺ concentration in the endoplasmic reticulum due to the compound was measured.

(1) Construction of expression plasmids The cDNAs of RyR1 and RyR2 are cloned from rabbit skeletal muscle and mouse myocardium by the PCR method and used as a hygromycin-resistant expression vector (pcDNA5 / FRT / TO) for Flp-In T-REX. Incorporated. This vector is a tetracycline-induced expression type (Tet-On) and induces RyR expression in the presence of doxycycline. R-CEPIA1er was incorporated into pCMV / myc / ER, an expression vector with neomycin resistance. This vector constitutively expresses R-CEPIA1er.

(2) HEK293 cells As HEK293 cells, Flp-In T-REX 293 cells corresponding to the Tet-On-induced system were used. Cells were cultured in a CO ₂ incubator at 37 ° C.

(3) Introduction of R-CEPIA1er (described in Non-Patent Document 13) and disease mutant RyR gene into HEK293 cells The disease mutant RyR gene is introduced into Flp-In T-REX 293 cells by the lipofection method to obtain hyglomycin. A stable expression strain was established by culturing in the added medium for 10 to 14 days. By introducing the R-CEPIA1er expression vector into this disease mutant RyR stable expression strain by the lipofection method and culturing in a medium supplemented with G418, a stable expression strain that double-expresses the disease mutant RyR gene and R-CEPIA1er can be obtained. Established.

(4) Introduction of R-CEPIA1er and wild-type RyR gene into HEK293 cells The wild-type RyR gene was introduced into Flp-In T-REX 293 cells by the lipofection method and cultured in a medium supplemented with hyglomycin for 10 to 14 days. By doing so, a stable expression strain was established. By introducing the R-CEPIA1er expression vector into this disease mutant RyR stable expression strain by the lipofection method and culturing in a medium supplemented with G418, a stable expression strain that double-expresses the wild-type RyR gene and R-CEPIA1er is established. did.

(5) Measurement of Ca ²⁺ concentration in fluorescent vesicles Cultured cells expressing disease mutant RyR and R-CEPIA1er and cultured cells expressing wild-type RyR and R-CEPIA1er were seeded on 96-well plates, respectively, and 37 The cells were cultured at ° C for 24 hours. The medium was replaced with a medium supplemented with doxycycline and cultured for another 24 hours to induce the expression of RyR. The medium was replaced with Krebs solution prior to measurement. Fluorescence measurement was performed by FlexStation. R-CEPIA1er was excited at 560 nm to obtain fluorescence at 610 nm. Fluorescence was acquired every 10 seconds for 300 seconds, and the following test compounds or control compounds were added at various concentrations 60 seconds after the start, and the Ca ²⁺ concentration in the fluorescent vesicles was measured.

(6) Compounds to be tested The compounds used in the test are as follows.
Compounds according to the present invention Compound 2: 1,4-dihydro-1-ethyl-4-oxoquinoline-3-carboxylic acid,
Compound 4: 1,4-dihydro-1-butyl-4-oxoquinoline-3-carboxylic acid,
Compound 8: 1,4-dihydro-1-octyl-4-oxoquinoline-3-carboxylic acid,
Compound 13: 1,4-dihydro-6-methyl-1-octyl-4-oxoquinoline-3-carboxylic acid,
Compound 15: 1,4-dihydro-7-methyl-1-octyl-4-oxoquinoline-3-carboxylic acid,
Compound 17: 1,4-dihydro-6-methoxy-1-octyl-4-oxoquinoline-3-carboxylic acid,
Compound 19: 1,4-dihydro-7-methoxy-1-octyl-4-oxoquinoline-3-carboxylic acid,
Compound 20: 5-ethyl-8-oxo- [1,3] dioxolo [4,5-g] quinoline-7-carboxylic acid (oxophosphate), and compound 29: 5-octyl-8-oxo- [1, 3] Dioxolo [4,5-g] Quinoline-7-carboxylic acid

-Control compound: 1,4-dihydro-4-oxoquinoline-3-carboxylic acid (DOCA)

Note that (2), (4), (8), (13), (15), (17), (19), (20) and (29) in FIGS. 1 to 6 indicate the above compound numbers.

(7) Results The results are shown in FIGS. 1 to 4.
FIG. 1 is a comparison of the RyR1 inhibitory activity of oxolinic acid (Compound 20) and DOCA, which is a control compound. Compared with DOCA, oxolinic acid improved RyR1 inhibitory activity.
FIG. 2 shows that the RyR1 inhibitory effect was improved by substituting an alkyl group for the nitrogen atom of DOCA as a control compound.
FIG. 3 shows the activity of a compound having a substituent at the 6-position or 7-position, using 1,4-dihydro-1-octyl-4-oxoquinoline-3-carboxylic acid (Compound 8, NC ₈ ) as a control compound. It is shown that the RyR1 inhibitory activity was improved particularly in the compound having a substituent at the 7-position.
FIG. 4 shows that Compound 29 has excellent RyR1 inhibitory activity against dandlorene, which is known as a therapeutic agent for malignant hyperthermia.

Pharmacological test 2 Ryanodine binding test The compound of the present invention was tested for its inhibitory activity against the malignant hyperthermia mutant RyR1 by the ryanodine binding method. The test method was carried out with reference to Non-Patent Document 10, and the degree of decrease in ryanodine binding in the group to which compound 29 was added was measured with respect to the control group.

(1) Preparation of microsomes HEK cells expressing the disease mutant RyR1 were disrupted with a nitrogen gas cell disruptor. After removing the nuclear fraction by low-speed centrifugation, ultracentrifugation was performed. The precipitate was suspended in buffer, ultracentrifuged, and then resuspended to obtain a microsome fraction.

(2) Ryanodine-conjugated microsomes were subjected to a reaction with tritium-labeled ryanodine in reaction solutions having various Ca ²⁺ concentrations at 37 ° C. for 2 hours. Compound 29 was added at a final concentration of 10 μM. The reaction solution was filtered through a glass filter treated with polyethyleneimine to separate ryanodine bound to RyR1.

(3) Results The results are shown in FIG. Compound 29 reduced the ryanodine binding of the disease mutant RyR1, especially in the low Ca ²⁺ concentration region. This indicates that compound 29 suppresses RyR1.

Pharmacological test 3 Animal test The therapeutic effect of the compound of the present invention on RyR1 malignant hyperthermia mutation-introduced mice was tested by the following method. The test method was to measure the increase in body temperature of mice caused by isoflurane anesthesia.

(1) Creation of RyR1 Malignant Hyperthermia Mutation-Introduced Mouse A knock-in mouse in which the RyR1 amino acid mutation (Arg2508Cys) identified in a malignant hyperthermia patient was introduced into a mouse was prepared. A targeting vector for substituting arginine (Arg) at position 2509 of mouse RyR1 with cysteine (Cys) was designed, and genome editing was performed on fertilized mouse eggs by CRISPR / Cas9. The born mice were screened to obtain founder mice into which the target mutation was introduced, and these were crossed with wild-type mice to obtain F1 heterozygous mice.

(2) Preparation of Solution of Compound 29 Compound 29 was dissolved in a 0.15 M aqueous sodium chloride solution and a 3 mM aqueous sodium hydroxide solution to prepare a 1 mg / mL solution.

(3) Isoflurane anesthesia test Wild-type or mutant mice were anesthetized with 2% isoflurane and then maintained with 1% isoflurane. The mouse rectal temperature was continuously measured with a digital thermometer (Tateyama Kagaku Kogyo, thermistor high-precision temperature data recording device K730) and recorded every 5 minutes. The solution of compound 29 was administered intraperitoneally to 10 mg / kg 10 minutes before the test.

(4) Results and discussion The results are shown in FIG. In wild-type mice (○), rectal temperature gradually increased for 30 minutes after isoflurane anesthesia, but did not die. Mutant mice (●) all died due to general muscle contracture between 26 and 33 minutes after the rectal temperature rose sharply 15 minutes after anesthesia (+ mark in the figure). On the other hand, the mutant mouse (▽) to which compound 29 was pre-administered suppressed the increase in body temperature and did not die. This result shows a remarkable preventive effect of compound (X) on malignant hyperthermia model mice.

Claims (9)

General formula (I)
[In the formula, R ¹ represents a C1-12 alkyl group, a C2-12 alkenyl group or a C2-12 alkynyl group, and the alkyl group, the alkenyl group and the alkynyl group are a C3-12 carbocycle or a 4- to 12-membered complex. The rings may be substituted,
X indicates CR ⁴ or N,
R ^{2, R 3} and ^{R 4} are each independently a hydrogen atom, Cl to 6 alkyl, C2-6 alkenyl, C2-6 alkynyl, C1 -3 fluoroalkyl group, Cl to 6 alkoxy group, OR ¹⁰ (in the formula, R ¹⁰ represents a hydrogen atom or a phenyl group), NR ¹¹ R ¹² (in the formula, R ¹¹ and R ¹² each independently represent a hydrogen atom, a C1-6 alkyl group or a phenyl group). ), COR ¹³ (in the formula, R ¹³ is a hydroxyl group, OR ²⁰ (in the formula, R ²⁰ represents a hydrogen atom, a C1-6 alkyl group or a phenyl group) or NR ²¹ R ²² (in the formula, R ²¹ and R ²² independently indicate a hydrogen atom, a C1-6 alkyl group or a phenyl group)), a cyano group, a nitro group, a halogen atom, a C1 to 3 fluoride alkoxy group, and a C1 to 6 alkylthio group. , Cl to 6 alkylthio group, C3-12 indicates a carbon ring or a 4-12 membered heterocyclic ring, ^{R 2} and ^{R 3} may indicate a C3~5 alkylene group or C1~3 alkylenedioxy group together ,
Y is COR ³⁰ (R ³⁰ is a hydroxyl group, OR ⁴⁰ (in the formula, R ⁴⁰ indicates a C1-6 alkyl group or a phenyl group) or NR ⁴¹ R ⁴² (in the formula, R ⁴¹ and R ⁴² are independent of each other). , Hydrogen atom, C1-6 alkyl group or phenyl group.) Is shown.). ]
A therapeutic or prophylactic agent for a ryanodine receptor-related disease containing the compound represented by (1) or a pharmaceutically acceptable salt thereof as an active ingredient. Claim that in the compound represented by the general formula (I), X is CH, R ¹ is a C4 to 10 alkyl group, R ² is a hydrogen atom, and R ³ is a C 1 to 4 alkyl group. 1 Therapeutic or prophylactic agent according to the above. The therapeutic or prophylactic agent according to claim 1 or 2, wherein the ryanodine receptor is RyR1. The therapeutic or prophylactic agent according to any one of claims 1 to 3, wherein the ryanodine receptor-related disease is a disease selected from the group consisting of malignant hyperthermia and central core disease. A ryanodine receptor inhibitor containing the compound represented by the general formula (I) according to claim 1 or a pharmaceutically acceptable salt thereof as an active ingredient. The inhibitor according to claim 5, wherein the ryanodine receptor is RyR1. General formula (IA)
[In the formula, R ^1A represents a C1-12 alkyl group, a C3-6 cycloalkyl-C1-4 alkyl group or a phenyl-C1-4 alkyl group.
X ^A represents CR ^4A or N, R ^4A represents a hydrogen atom, a C1-4 alkyl group or a C1-4 alkoxy group.
One of R ^2A and R ^3A is a hydrogen atom, C1 to 4 alkyl group or C1 to 4 alkoxy group, and the other is hydrogen atom, C1 to 4 alkyl group, C1 to 4 alkoxy group, cyano group, halogen atom and trihalomethyl. A group, a trihalomethyloxy group or a methylthio group, or R ^2A and R ^3A together to indicate a C3-5 alkylene group or an -O-CH _2- O- group (provided that (i) R ^2A and When both R ^3A are hydrogen atoms, R ^1A shows a cyclohexylmethyl group,
(Ii) When R ^1A is an n-propyl group, R ^2A is not a methyl group or a methoxy group,
(Iii) R ^1A is not an octyl group when R ^2A and R ^3A together indicate an -O-CH _2- O- group)] or a pharmaceutically acceptable salt thereof. In the compound represented by the general formula (IA) according to claim 7.
One of R ^2A and R ^3A is a hydrogen atom, a C1-4 alkyl group or a C1-4 alkoxy group, and the other is a C1-4 alkyl group, a C1-4 alkoxy group, a halogen atom, a cyano group or a trihalomethyl group. Or, R ² and R ³ together represent an -O-CH _2- O- group (where (i) when R ^1A is an n-propyl group, R ^2A is not a methyl or methoxy group,
(Ii) R ^1A is not an octyl group when R ^2A and R ^3A together indicate an -O-CH _2- O- group)]
The compound according to claim 7 or a pharmaceutically acceptable salt thereof. (1) 1-Cyclohexylmethyl-1,4-dihydro-4-oxoquinoline-3-carboxylic acid,
(2) 1,4-dihydro-6-methyl-1-octyl-4-oxoquinoline-3-carboxylic acid,
(3) 1,4-Dihydro-7-methyl-1-propyl-4-oxoquinoline-3-carboxylic acid,
(4) 1,4-dihydro-7-methyl-1-octyl-4-oxoquinoline-3-carboxylic acid,
(5) 1,4-dihydro-7-methyl-1-octyl-4-oxoquinoline-3-carboxylic acid,
(6) 1,4-Dihydro-7-methoxy-4-oxo-1-propylquinoline-3-carboxylic acid,
(7) 1,4-Dihydro-7-methoxy-1-octyl-4-oxoquinoline-3-carboxylic acid,
(8) 1,4-Dihydro-1-propyl-4-oxo-7-trifluoromethylquinoline-3-carboxylic acid,
(9) 1,4-dihydro-1-octyl-4-oxo-7-trifluoromethylquinoline-3-carboxylic acid,
(10) 5-decyl-8-oxo- [1,3] dioxolo [4,5-g] quinoline-7-carboxylic acid,
(11) 7-Methoxy-1-propyl-1,4-dihydro-4-oxo-1,8-naphthylidine-3-carboxylic acid,
(12) 1,4-Dihydro-7-methoxy-1-octyl-4-oxo-1,8-naphthylidine-3-carboxylic acid,
(13) 1-decyl-1,4-dihydro-7-methoxy-4-oxo-1,8-naphthylidine-3-carboxylic acid,
(14) 1,4-dihydro-6,7-dimethoxy-1-octyl-4-oxoquinoline-3-carboxylic acid,
(15) 1-decyl-1,4-dihydro-6,7-dimethoxy-4-oxoquinoline-3-carboxylic acid,
(16) 1-propyl-4-oxo-cyclopenta [g] quinoline-3-carboxylic acid,
(17) 1-octyl-4-oxo-cyclopenta [g] quinoline-3-carboxylic acid,
(18) 1-propyl-1,4-dihydro-7-trifluoromethyloxy-4-oxoquinoline-3-carboxylic acid,
(19) 1-propyl-1,4-dihydro-7-trifluoromethyloxy-4-oxoquinoline-3-carboxylic acid,
(20) 1,4-dihydro-8-methoxy-1-octyl-4-oxoquinoline-3-carboxylic acid,
(21) 7,8-dimethoxy-1,4-dihydro-1-octyl-4-oxoquinoline-3-carboxylic acid, and (22) 1,4-dihydro-7-methylthio-1-octyl-4-oxo The compound according to claim 7, or a pharmaceutically acceptable salt thereof, selected from the group consisting of quinoline-3-carboxylic acid.