WO2019082940A1

WO2019082940A1 - Ryanodine receptor inhibitor

Info

Publication number: WO2019082940A1
Application number: PCT/JP2018/039542
Authority: WO
Inventors: 尚村山; なごみ呉林國廣; 影近　弘之; 修一森; 磨里湯浅
Original assignee: 学校法人順天堂; 国立大学法人東京医科歯科大学
Priority date: 2017-10-25
Filing date: 2018-10-24
Publication date: 2019-05-02
Also published as: JP7282332B2; JPWO2019082940A1

Abstract

The purpose of the present invention is to provide a medicament containing as an active ingredient a compound having ryanodine inhibitory activity, in particular, RyR1 inhibitory activity. Provided is an agent for treating or preventing ryanodine receptor-related diseases, the agent containing as an active ingredient a compound represented by formula (I) (all of the symbols in the formula are defined as in the description) or a pharmaceutically acceptable salt thereof.

Description

Ryanodine receptor inhibitor

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 cytoplasmic side to release Ca ²⁺ into the cytosol (Ca ²⁺ -induced Ca ²⁺ release, CICR). So it acts as a positive feedback mechanism, and small amounts of Ca ²⁺ in the cytosol near the channel cause more Ca ²⁺ release from SR.

As RyR, subtypes of RyR1 mainly expressed in skeletal muscle, RyR2 mainly expressed in cardiac muscle, and RyR3 widely expressed in brain but widely expressed in brain are known. RyR gene mutations cause abnormally enhanced CICR activity and are considered to be the cause of various diseases. For example, RyR1-related diseases include hyperthermia malignancy (MH) (eg, increase in body temperature due to inhaled anesthetic during surgery, skeletal muscle contracture, etc.), central core disease (CCD) (muscle of limbs designated as intractable disease) And the like)) (Non-Patent Document 1). Moreover, as RyR2-related diseases, catecholamine-induced polymorphic ventricular tachycardia (CPVT) (motor arrhythmia), arrhythmogenic right ventricular cardiomyopathy (ARVC) (right ventricular dysplasia, arrhythmia induction), idiopathic ventricular Fibrillation (IVF) (ventricular fibrillation induction at rest) and the like can be mentioned (Non-patent Document 2). Furthermore, as a disease resulting from the secondary occurrence of dysregulation of RyR due to a major factor other than RyR, malignant syndrome (side effect of a drug for treating schizophrenia), muscular dystrophy, Alzheimer's disease, Huntington's disease, trauma Sexual brain injury etc. (Non-patent documents 3 to 8).

The cause of these RyR related diseases is thought to be the abnormal enhancement of CICR activity by RyR gene mutation, but there is no effective therapeutic agent at this time. For example, dantrolene is regarded as the only therapeutic agent for malignant hyperthermia (Non-patent Document 9).

Japanese Patent Publication No. 51-18440 International Publication No. 2010/064701 brochure

However, dantrolene is unsuitable for long-term administration due to strong side effects, and its administration method is limited due to its low water solubility. In addition, dantrolene has no application as a CCD therapeutic agent. Therefore, an object of the present invention is to provide a medicament having ryanodine receptor inhibitory activity, particularly inhibitory activity against RyR1, and effective for ryanodine receptor related diseases.

Therefore, as a result of various investigations in view of the relationship between ryanodine receptor and CICR which is an essential step for muscle contraction, the present inventors cultured cells expressing a fluorescent endoplasmic reticulum Ca ²⁺ indicator and disease-altered 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 the screening of CICR activity inhibitors, ie, ryanodine receptor related disease therapeutic agents can be performed by comparing the concentrations, and a patent application was filed earlier (Japanese Patent Application No. 2016-113147, and Non-patent Document 10). And 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 a preventive or therapeutic agent for ryanodine receptor related diseases As a result, the present invention has been completed.

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

[Wherein, 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 have a C3-12 carbon ring, or a 4 to 12-membered complex] The ring may be substituted,
X represents CR ⁴ or N,
R ² , R ³ and R ⁴ are each independently a hydrogen atom, a C1-6 alkyl group, a C2-6 alkenyl group, a C2-6 alkynyl group, a C1 to 3 fluoroalkyl group, a C1 to 6 alkoxy group, OR ¹⁰ (wherein, R ¹⁰ represents a hydrogen atom or a phenyl group), NR ¹¹ R ¹² (wherein, R ¹¹ and R ¹² each independently represent a hydrogen atom, a C1-6 alkyl group or a phenyl group) ), COR ¹³ (wherein, R ¹³ is a hydroxyl group, OR ²⁰ (wherein, R ²⁰ is a hydrogen atom, a C1-6 alkyl group or a phenyl group)) or NR ²¹ R ²² (wherein, R ²¹ and R ²² each independently represents a hydrogen atom, a C 1 to 6 alkyl group or a phenyl group)), a cyano group, a nitro group, a halogen atom, a C 1 to 3 fluorinated alkoxy group, a C 1 to 6 alkyl And R ² and R ³ may be taken together to represent a C 3-5 alkylene group or a C 1-3 alkylenedioxy group, which is a thio group, a C 3-12 carbon ring or a 4 to 12-membered heterocyclic ring;
Y is COR ³⁰ (R ³⁰ is a hydroxyl group, OR ⁴⁰ (wherein R ⁴⁰ is a C1-6 alkyl group or a phenyl group)) or NR ⁴¹ R ⁴² (wherein R ⁴¹ and R ⁴² are each independently. , A hydrogen atom, a C.sub.1-6 alkyl group or a phenyl group is shown). ]
A therapeutic or preventive agent for a ryanodine receptor related disease, which comprises the compound represented by or the pharmaceutically acceptable salt thereof as an active ingredient.

[2] In the compound represented by the general formula (I) described in the preceding paragraph [1], X is CH, R ¹ is a C 6-10 alkyl group, R ² is a hydrogen atom, R ³ is C 1 The therapeutic or prophylactic agent according to the above [1], which is a -4 alkyl group.

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

[4] The therapeutic or prophylactic agent according to any one of the above-mentioned [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 ryanodine receptor, which comprises the compound represented by the general formula (I) according to the preceding paragraph [1] or a pharmaceutically acceptable salt thereof as an active ingredient.

[6] The inhibitor according to the above-mentioned [5], wherein the ryanodine receptor is RyR1.

[7] General Formula (IA)

[Wherein, 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, and R ^4A represents a hydrogen atom, a C 1-4 alkyl group or a C 1-4 alkoxy group,
One of R ^2A and R ^3A is a hydrogen atom, a C 1-4 alkyl group or a C 1-4 alkoxy group, and the other is a hydrogen atom, a C 1-4 alkyl group, a C 1-4 alkoxy group, a cyano group, a halogen atom, trihalomethyl Group, a trihalomethyloxy group or a methylthio group, or R ^2A and R ^3A together represent a C 3-5 alkylene group or an -O-CH ₂ -O- group (however, (i) R ^2A and When R ^3A is both hydrogen atoms, R ^1A represents a cyclohexylmethyl group,
(Ii) when R ^1A is n-propyl, R ^2A is not methyl or methoxy;
(Iii) When R ^2A and R ^3A are taken together to represent an —O—CH ₂ —O— group, R ^1A is not an octyl group) or a pharmaceutically acceptable salt thereof.

[8] In the compound represented by the general formula (IA) described in the preceding paragraph [7],
One of R ^2A and R ^3A is a hydrogen atom, and the other is a C 1-4 alkyl group, a C 1-4 alkoxy group, a halogen atom, a cyano group or a trihalomethyl group, or R ² and R ³ are together. shows a _-O-CH 2 -O- group Te (where, ⁽ⁱ⁾ ^{R 2A} when ^{R 1A} is n- propyl group is not a methyl group or a methoxy group,
(Ii) R ^1A is not an octyl group when R ^2A and R ^3A together represent an -O-CH ₂ -O- group)]
A 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] dioxoro [4,5-g] quinoline-7-carboxylic acid,
(11) 1,4-dihydro-7-methoxy-1-propyl-4-oxo-1,8-naphthyridine-3-carboxylic acid,
(12) 1,4-dihydro-7-methoxy-1-octyl-4-oxo-1,8-naphthyridine-3-carboxylic acid,
(13) 1-decyl-1,4-dihydro-7-methoxy-4-oxo-1,8-naphthyridine-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 above item [7] or a pharmaceutically acceptable salt thereof, which is 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 is useful for the prevention or treatment of skeletal muscle diseases because it exhibits the inhibitory activity of the ryanodine receptor, particularly the RyR1 receptor.

The RyR1 inhibitory activity of oxophosphoric acid and 1,4-dihydro-4-oxoquinoline 3-carboxylic acid (DOCA) is shown. The RyR1 inhibitory activity of the compound used by this invention is shown. The RyR1 inhibitory activity of the compound used by this invention is shown. The RyR1 inhibitory activity of the compound used by this invention is shown. The RyR1 binding activity of the compound used by this invention is shown. The therapeutic effect on RyR1 malignant hyperthermia mutation-introduced mice is shown.

The active ingredient in the therapeutic or preventive agent for ryanodine receptor related diseases of the present invention is a compound represented by the general formula (I) or a pharmaceutically acceptable salt thereof. The compounds represented by the general formula (I) include oxophosphoric acid and quinoline-3-carboxylic acid derivative compounds. Oxolinic acid is a quinolone antibacterial agent developed in the 1970s (Patent Document 1). Oxophosphate inhibits gyrase of bacterial DNA and is currently used mainly as an antimicrobial agent for animals. In addition, as a compound whose structure is similar to oxophosphoric acid, studies of structure-activity relationship with respect to the antimicrobial activity of 1,4-dihydro-4-oxoquinoline-3-carboxylic acid (DOCA) derivatives have been conducted (eg, non-patented) 11 and 12). These compounds are known to have antibacterial activity, but none are known to have ryanodine receptor inhibitory activity. In addition, the compounds represented by the general formula (IA) are novel compounds which have 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 alkyl group having 3 to 12 carbon atoms. Examples of C1-12 alkyl groups include methyl, ethyl, n-propyl, n-butyl, n-pentyl, n-hexyl, n-heptyl, n-octyl and n-nonyl Group, n-decyl group, n-undecyl group, n-dodecyl linear alkyl group, isopropyl group, isobutyl group, t-butyl group, s-butyl group, isopentyl group, 1-ethylpropyl group, 3 Branched chains such as -methylbutyl, 2,2-dimethylpropyl, isohexyl, 3-ethylbutyl, 3,3-dimethylbutyl, isoheptenyl, isooctyl, isononyl, isodecyl, isoundecyl and isododecyl groups Alkyl groups are 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 alkenyl group having 3 to 12 carbon atoms. Examples of C2-12 alkenyl groups include ethenyl group, n-propenyl group, n-butenyl group, n-pentenyl group, n-hexenyl group, n-heptanyl group, n-octanyl group, n-nonenyl group, n -Decenyl group, n-undecenyl group, linear alkenyl group of n-dodecenyl group, isopropenyl group, isobutenyl group, s-butenyl group, isopentenyl group, 1-ethylpropenyl group, 3-methylbutenyl group, 2, Branched alkenyl groups such as 2-dimethylpropenyl group, isohexenyl group, 3-ethylbutenyl group, 3, 3-dimethylbutenyl group, isoheptenyl group, isooctenyl group, isonononyl group, isodecenyl, isoundecenyl group, isododecenyl group, etc. It 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-heptynyl group, n-octynyl group, n-nonynyl group, n -Decynyl group, n-undecynyl group, linear alkynyl group of n-dodecynyl group, isobutynyl group, s-butynyl group, isopentynyl group, 1-ethylpropynyl group, 3-methylbutynyl group, 2,2-dimethylpropynyl group And branched alkynyl groups such as isohexynyl group, 3-ethylbutynyl group, 3,3-dimethylbutynyl group, isoheptynyl group, isoheptynyl group, isononynyl group, isononynyl group, isodecynyl, isoundecynyl group, isododecynyl group and the like.

In the present specification, “C3-12 carbocyclic ring” is a saturated hydrocarbon (cycloalkane) such as cyclopropane, cyclobutane, cyclopentane, cyclohexane, cycloheptane, cyclooctane, cyclononane, cyclodecane, bicycloheptane, bicyclooctane, bicyclononane Carbocyclic rings such as bicyclo rings such as cyclobutene, cyclopentene, cyclohexene, cycloheptene, cyclooctene, cyclononene, cyclodecene cycloalkenes, aromatic rings such as benzene, naphthalene, indane, indene and the like; It can be mentioned.

In the present specification, the "4 to 12-membered heterocyclic ring" includes "4 to 12-membered saturated heterocyclic ring" and "4 to 12-membered unsaturated heterocyclic ring".
The “4- to 12-membered saturated heterocycle” is a saturated monocyclic or bicyclic or more heterocycle having a heteroatom selected from nitrogen atom, oxygen atom and sulfur atom, and examples thereof include morpholine, 1- Pyrrolidine, piperidine, piperazine, tetrahydrofuran, tetrahydropyran, tetrahydrothiophene, thiazolidine, oxazolidine and the like can be mentioned.

In the present specification, the "4 to 12-membered unsaturated heterocyclic ring" is a fully or partially unsaturated monocyclic or bicyclic ring having a heteroatom selected from nitrogen, oxygen and sulfur atoms. And heterocyclic rings such as imidazoline, thiophene, furan, pyrrole, oxazole, isoxazole group, thiazole, isothiazole group, thiadiazole group, pyrazole, triazole, tetrazole, pyridine, pyrazine, pyrimidine, pyridazine, indole, isoindole, Indazole group, triazolopyridine, benzoimidazole, benzoxazole, benzothiazole group, benzothiophene, benzofuran, purine, quinoline, isoquinoline, quinazoline, quinoxaline, methylenedioxybenzene, ethylenedioxybenzene, dihydroben Fran, and the like.

In the present specification, the “C1-6 alkyl group” refers to a linear alkyl group having 1 to 6 carbon atoms or a branched alkyl group having 3 to 6 carbon atoms. Examples of the C1-6 alkyl group include methyl group, ethyl group, n-propyl group, n-butyl group, n-pentyl group, linear alkyl group of n-hexyl group, isopropyl group, isobutyl group, 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. And a branched alkyl group of

In the present specification, the "C1-4 alkyl group" refers to a linear alkyl group having 1 to 4 carbon atoms or a branched alkyl group having 3 to 4 carbon atoms. Examples of the C1-4 alkyl group include a methyl group, an ethyl group, an n-propyl group, a linear alkyl group of n-butyl group, an isopropyl group, an isobutyl group, a t-butyl group and an s-butyl group A branched alkyl group can be mentioned.

In the present specification, the “C2-6 alkenyl group” refers to a linear alkenyl group having 2 to 6 carbon atoms or a branched alkenyl group having 3 to 6 carbon atoms. Examples of the C2-6 alkenyl group include ethenyl group, n-propenyl group, n-butenyl group, n-pentenyl group, linear alkenyl group of n-hexenyl group, isopropenyl group, isobutenyl group, s- Examples thereof include branched alkenyl groups such as butenyl group, isopentenyl group, 1-ethylpropenyl group, 3-methylbutenyl group and 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, linear alkynyl group of n-hexynyl group, isobutynyl group, s-butynyl group, isopentinyl group And branched alkynyl groups such as 1-ethylpropynyl group, 3-methylbutynyl group, 2,2-dimethylpropynyl group and isohexynyl group.

In the present specification, as the "C.sub.1-3 trifluorinated alkyl group", fluoromethyl, difluoromethyl, trifluoromethyl, fluoroethyl, difluoroethyl, trifluoroethyl, trifluoroethyl, fluoropropyl, difluoropropyl, trifluoropropyl, hexafluoropropyl And the like.

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

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

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

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

In the present specification, examples of the "C1-C3 fluorinated alkoxy group" include fluoromethyloxy, difluoromethyloxy, trifluoromethyloxy, fluoroethyloxy, difluoroethyloxy, trifluoroethyloxy, fluoropropyloxy, difluoropropyloxy , Trifluoropropyloxy, hexafluoropropyloxy and the like.

In the present specification, the "C1-3 alkylenedioxy group" refers to a methylenedioxy, ethylenedioxy or propylenedioxy group.

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

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

Hereafter, the preferable aspect of a compound represented by general formula (I) is 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.

As R ¹ , a C1-12 alkyl group, a C2-12 alkenyl group or a C2-12 alkynyl group is preferable, a C1-12 alkyl group is more preferable, a C2-12 alkyl group is more preferable, and a C4-10 alkyl group is more preferable Preferred is a C6-10 alkyl group.

As R ² , a hydrogen atom, a C1-6 alkyl group and a C1-6 alkoxy group are preferable, and a hydrogen atom is more preferable.

As R ³ , a hydrogen atom, a C 1-6 alkyl group and a C 1-6 alkoxy group are preferable, and a C 1-4 alkyl group and a C 1-4 alkoxy group are more preferable.

Also, R ² and R ³ may be combined to form a C 1-3 alkylenedioxy group.

In the compounds represented by the general formula (I), X is CH, R ¹ is a C 4-10 alkyl group, R ² is a hydrogen atom, R ³ is a C 1-4 alkyl group or C 1-4 alkoxy Compounds which are groups are preferred.

The preferable aspect of a compound represented by general formula (IA) is shown.

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

As R ^1A , a C1-12 alkyl group is preferable, and a C6-10 alkyl group is 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.

In addition, R ^2A and R ^3A may be combined to form a C 3 to 5 alkylene group or a C 1 to 3 alkylenedioxy group.

In the compound represented by the general formula (I) used in the present invention, specifically,
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 (oxophosphoric acid), and compound 29: 5-decyl-8-oxo- [1,1 3] Dioxoro [4,5-g] quinoline-7-carboxylic acid is preferred,
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 (oxophosphoric acid), and compound 29: 5-decyl-8-oxo- [1,1 3] Dioxoro [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 (oxophosphoric acid) and compound 29: 5-decyl-8-oxo- [1,3 Further preferred is dioxolo [4,5-g] quinoline-7-carboxylic acid.

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

The compounds represented by the general formulas (I) and (IA) and pharmaceutically acceptable salts thereof may contain solvates (including hydrates) and crystal polymorphs, but may be isolated It may be a crystalline form or a mixture of multiple crystalline forms, both of which are included. It may also be in amorphous form.

The compounds represented by the general formulas (I) and (IA) or pharmaceutically acceptable salts thereof are substituted using characteristics which are themselves known or which are based on the basic skeleton or the kind of their substituents. It can be manufactured by applying various known synthetic methods for group introduction and functional group conversion. For example, using 1,4-dihydro-4-oxoquinoline-3-carboxylic acid (DOCA, CAS number: 13721-01-2) as a starting material, a reaction for introducing a substituent is performed by a known method, or Patent Document 2 It can manufacture according to the method as described in the nonpatent literature 11,12. For example, oxophosphoric acid is a known compound assigned CAS number: 14698-29-4.

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

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

Step (1)
Among the compounds represented by the general formula (I), a compound in which Y is a group other than a COOH group, that is, a compound represented by the general formula (I-1) is a compound represented by the general formula (II) It can be produced by N-alkylation reaction which is a reaction with the compound represented by (III). This reaction can be carried out in the presence or absence of a solvent in the presence of an alkaline agent or a strong base.
The compound represented by the formula (III) is a halogenated hydrocarbon, and methane chloride, bromide, iodide, methane ethane, bromide n-propane, bromide n-propane, bromide isopropane, 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 halide, benzyl bromide and allyl bromide can be used.
Examples of the alkaline agent include inorganic alkaline compounds such as sodium hydrogencarbonate, sodium carbonate, potassium carbonate, calcium carbonate, cesium carbonate, sodium hydroxide, potassium hydroxide and the like.
Examples of strong bases include sodium hydride and potassium hydride.
The solvent depends on the alkyl halide and the alkalizing agent used, but a solvent generally used for organic synthesis, for example, alcohol solvents such as methanol, ethanol and 2-propanol, ether solvents such as diethyl ether, tetrahydrofuran and dioxane, Aromatic solvents such as benzene, toluene and xylene, halogen solvents such as chloroform and dichloromethane, acetonitrile, DMF, DMSO, N-methylpyrrolidone and the like, water, or a mixture thereof can be used.
Even when a compound in which Y ¹ group of the compound represented by the general formula (II) is replaced with COOH group is used as a raw material, N-alkylation and esterification proceed simultaneously by the reaction of step (1) The compound shown by I-1) can be produced.

Step (2)
Among the compounds represented by the general formula (I), a compound in which Y is a carboxyl group (a compound represented by the general formula (I-2)) is a compound represented by the general formula (I-1) obtained by the step (1) Can be obtained by subjecting the compound represented by) 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 a base (eg, sodium hydroxide etc.) or catalytic reduction (eg, in the presence of 10% palladium-carbon catalyst) ) Can be implemented. The reaction can be carried out at room temperature to reflux temperature for 1 to 24 hours.
As the solvent, solvents generally used in organic synthesis, for example, alcohol solvents such as methanol, ethanol and 2-propanol, ether solvents such as diethyl ether, tetrahydrofuran and dioxane, aromatic solvents such as benzene, toluene and xylene Halogen solvents such as chloroform and dichloromethane, acetonitrile, DMF, DMSO, N-methyl pyrrolidone and the like, water, or a mixture of these can be used.

Step (3)
Among the compounds represented by the general formula (I), Y is COOR ⁴⁰ (R ⁴⁰ has the same meaning as described above), 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 by the above step (2) to an esterification or amidation reaction can do.
The esterification reaction and the amidation reaction are, for example,
(A) Method using an acid halide,
(B) a method using a mixed acid anhydride,
(C) It can carry out by the method of using a condensing agent.

If these methods are explained concretely,
(A) The method of using an acid halide can be carried out, for example, by using a carboxylic acid in an organic solvent (chloroform, dichloromethane, diethyl ether, tetrahydrofuran or the like) or without a solvent with an acid halide agent (oxalyl chloride, thionyl chloride or the like) The acid halide thus obtained is reacted at 20 ° C. to reflux temperature, and the resulting acid halide is reacted with an alcohol and an organic solvent (chloroform, dichloromethane, diethyl ether, tetrahydrofuran etc.) in the presence of a base (pyridine, triethylamine, dimethylaniline, dimethylaminopyridine, diisopropylethylamine etc.) ) At a temperature of about 0-40 ° C.). The reaction can also be carried out by reacting with an acid halide at about 0 to 40 ° C. using an aqueous alkaline solution (such as aqueous sodium bicarbonate or sodium hydroxide solution) in an organic solvent (such as dioxane or tetrahydrofuran).
(B) A method using a mixed acid anhydride is, for example, a base (pyridine, triethylamine, dimethylaniline, dimethylaminopyridine, diisopropyl) in an organic solvent (chloroform, dichloromethane, diethyl ether, tetrahydrofuran or the like) or without a solvent. Obtained by reacting with acid halide (pivaloyl chloride, tosyl chloride, mesyl chloride etc.) or acid derivative (ethyl chloroformate, isobutyl chloroformate etc.) in the presence of ethylamine etc.) at about 0 to 40 ° C. The reaction is carried out by reacting the mixed acid anhydride with an alcohol at about 0-40 ° C. in an organic solvent (chloroform, dichloromethane, diethyl ether, tetrahydrofuran or the like).
The method (C) using a condensing agent includes, for example, carboxylic acid and alcohol in an organic solvent (chloroform, dichloromethane, dimethylformamide, diethyl ether, tetrahydrofuran etc.) or without a base (pyridine, triethylamine, dimethylaniline, Condensation agent [1,3-dicyclohexylcarbodiimide (DCC), 1-ethyl-3- [3- (dimethylamino) propyl] carbodiimide (EDC), 1,1 ′ in the presence or absence of dimethylaminopyridine etc. -Carbonyldiimidazole (CDI), 2-chloro-1-methylpyridinium iodine, 1-propylphosphonic acid cyclic anhydride (PPA, etc.)], with or without 1-hydroxybenztriazole (HOBt), Conducted by reaction at 0 to 40 ° C Be
It is desirable that all of the reactions (A), (B) and (C) be carried out under an inert gas (for example, argon, nitrogen etc.) atmosphere under anhydrous conditions.

The compounds represented by the general formula (II) used as starting materials are known or known methods, for example, “Comprehensive Organic Transformations: A Guide to Functional Group Preparations 2nd Edition (Richard C. Larock, John Wiley & Sons Inc) , 1999) "can be easily manufactured. For example, it can be produced by the method shown in the following reaction scheme 2.

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

Step (4)
The reaction of the compound represented by the general formula (III) with the compound (methylene malonate ester) represented by the general formula (IV) can be carried out without solvent or in the presence of a solvent. As the solvent, water or an organic solvent (eg, ether solvents such as dioxane, tetrahydrofuran, diethyl ether, 1,2-dimethoxyethane, diethylene glycol dimethyl ether, ethylene glycol dimethyl ether, etc .; aromatic hydrocarbon solvents such as benzene, toluene, xylene etc. Lower alcohol solvents such as methanol, ethanol and isopropanol; polar solvents such as DMF, DMSO, hexamethylphosphoric acid triamide, and acetonitrile are used singly or as a mixture of two or more. The above reaction can be carried out under temperature conditions of usually room temperature to 200 ° C., preferably room temperature to 150 ° C., for any time of 1 to 30 hours.

Step (5)
The cyclization reaction starting from the compound represented by the general formula (V) can be carried out by heating in a solvent such as diphenyl ether as described in Patent Document 1. Moreover, it can also implement by heating the compound represented by general formula (V) also in absence of a solvent. The 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 known methods using known compounds as raw materials.

The compound represented by the general formula (I) or a pharmaceutically acceptable salt thereof has ryanodine receptor, particularly ryanodine 1 receptor (RyR1) inhibitory activity, and it is a skeletal muscle in animals such as mammals, especially 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 made into a pharmaceutical composition by being combined with a pharmaceutically acceptable carrier. As pharmaceutically acceptable carriers, known ones such as excipients, binders, buffers, thickeners, stabilizers, emulsifiers, dispersants, suspending agents, preservatives and the like can be used. It can be formulated by the usual methods.

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

The preparation for oral administration can be prepared according to a known method by incorporating additives commonly 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 polyvinyl pyrrolidone; disintegrants such as starch and carboxymethyl cellulose; magnesium stearate, Lubricants such as talc and the like can be mentioned. Parenteral administration can be administered as an injection, a preparation for rectal administration, a topical administration and the like, and among them, an injection is preferred.

Injections include, for example, sterile solutions or suspensions. These injections are produced, for example, by dissolving or suspending the compound of the present invention or a pharmaceutically acceptable salt thereof in water for injection by Japan Post. If necessary, an isotonicity agent such as sodium chloride; a buffer such as sodium dihydrogen phosphate or sodium monohydrogen phosphate; a solubilizing agent etc. may be blended. In addition, it can be used as an injectable preparation in a soluble form (powder filling, lyophilization), and in this case, it can be manufactured by an ordinary method by adding an excipient such as mannitol or lactose.

Suppository etc. are mentioned as a preparation for rectal administration. The 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 cocoa butter or macrogol, and pouring it into a mold for molding. Alternatively, the liquid or cream may be placed in a container for injection to give a preparation for rectal administration.

The topical administration preparations include solutions, eye drops, creams, ointments, gel preparations, sprays, powders and the like. The liquid preparation may 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 this thickener, gelatin, sodium hyaluronate, high molecular weight dextran, sodium alginate, sodium chondroitin sulfate and the like can be used. Eyedrops can be produced by adding a preservative, in addition to a buffer, pH adjuster, tonicity agent. Creams and ointments can be prepared using aqueous or oily bases such as water, liquid paraffin, vegetable oils (peanut oil, castor oil, etc.), macrogol and the like. The gel preparation may be gelatin, pectin, carrageenan, agar, tragacanth, alginate, cellulose ether (methylcellulose, sodium carboxymethylcellulose etc.), pectin derivative, polyacrylate, polymethacrylate, polyvinyl alcohol, polyvinyl pyrrolidone etc. Can be manufactured. A spray can be prepared by dissolving or suspending the compound of the present invention or a pharmaceutically acceptable salt thereof in water or the like, and then placing 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 can be produced by mixing it with a suitable excipient.

The dose of the compound of the present invention is appropriately determined depending on the individual case in consideration of the target disease or condition, the age, body weight, sex and the like of the administration subject. Usually, in the case of oral administration, the dose of the compound of the present invention per day for adults (body weight about 60 kg) is 0.01 to 100 mg, preferably 0.01 to 30 mg, more preferably 0.1 to 10 mg. Is administered once or in 2-4 divided doses. In the case of intravenous administration, 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 kg body weight, and 1 day 1 Administration is divided into multiple doses.

[Example]
EXAMPLES Hereinafter, the present invention will be specifically described by way of 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 type flask, 1,4-dihydro-4-oxoquinoline-3-carboxylic acid (200 mg, 1.06 mmol) and methyl iodide (450 mg, 3.18 mmol), N, N-dimethylformamide (3 mL) Solution was added and cesium carbonate (1.03 g) was further added, and the mixture was stirred at 50.degree. After 6 hours, chloroform and water were added to the reaction solution to separate it. 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)

In a 30 mL eggplant type flask, the compound (0.37 mmol) produced in Reference Example 1 was dissolved in methanol (3 mL). Thereto was added dropwise 1 M aqueous sodium hydroxide solution (2 mL) 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 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 needles (yield: 96%).
¹ H-NMR (500 MHz, CDCl ₃ ): δ 8.56 (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)
The following compounds were produced in the same manner as in Reference Example 1 → Synthesis Example 1, using the corresponding alkyl iodide compound instead of methyl iodide.

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 ): δ 15.0 (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 ): δ 14.95 (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 ): δ 14.96 (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 ): δ 14.99 (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 ): δ 14.89 (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 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
p-Toluidino methylene malonate diethyl

In a 30 mL eggplant type flask, p-toluidine (1.74 g, 16.2 mmol) was dissolved in ethanol (20 mL). While stirring the mixture, diethyl ethoxymethylene malonate (4.0 g, 16.3 mmol) was added dropwise, and the mixture was stirred at room temperature for 18 hours. After distilling off ethanol, the residue is partitioned between ethyl acetate and water, the organic layer is dried over anhydrous sodium sulfate and concentrated under reduced pressure, and the residue is 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

In a 200 mL two-necked flask, after heating diphenyl ether (50 mL) to 220 ° C., a solution of the compound (6.4 mmol) prepared in Reference Example 2 in 10 mL of diphenyl ether was added dropwise over 30 minutes. It stirred. The reaction solution was returned to room temperature, and 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 dry 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 then propyl iodide (0.86 mmol) was further added dropwise. After stirring for 9 hours at 55 ° C., the temperature was returned to room temperature, and the reaction was quenched by the addition of saturated aqueous ammonium chloride solution. Ethyl acetate and water were added and the mixture was separated, 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 ): δ 8.45 (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 type flask, the compound (0.32 mmol) prepared in Compound Reference Example 4 was dissolved in methanol (3 mL). 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 aqueous hydrochloric acid 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 needles (yield: 74%).
¹ H-NMR (500 MHz, CDCl ₃ ): δ 15.09 (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) to Synthesis Example 2 (28)
Reference Example 2 (if necessary, a corresponding compound was used instead of p-toluidine) → Reference Example 3 → Reference Example 4 (if necessary, corresponding alkyl iodide was used instead of ethyl iodide) → The following compounds were produced in the same manner as the method 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 ): 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 (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 ): 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 (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 ): 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 (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 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 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 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, 1 H), 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, 1 H), 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, 2 H), 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] dioxoro [4,5-g] quinoline-7-carboxylic acid (compound 29)

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-naphthyridine-3-carboxylic acid (compound 30)

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-naphthyridine-3-carboxylic acid (compound 31)

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-naphthyridine-3-carboxylic acid (compound 32)

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 (4 s, 3 H), 4.02 (s, 3 H), 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, 3 H), 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, 1 H), 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, 1 H), 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, 2 H), 3.08 (t, J = 7.6 Hz, 2 H), 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 H-NMR (400 MHz, CDCl ₃ ) δ 14.59 (s, 1 H), 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, 2 H), 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, 1 H), 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, 2 H), 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) .Composition 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, 3 H).
Synthesis example 2 (28)
1,4-Dihydro-7-methylthio-1-octyl-4-oxoquinoline-3-carboxylic acid (Compound 41)

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 for RyR1 The compounds of the present invention were tested for inhibitory activity against RyR1 by the following method. The test method is carried out with reference to Ca ²⁺ concentration ([Ca ²⁺ ] _ER ) in the endoplasmic reticulum of HEK 293 cells expressing a disease variant (malignant hyperthermia, central core disease) with reference to Non-patent document 10 The increase in endoplasmic reticulum Ca ²⁺ concentration by the compound was measured.

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

(2) HEK 293 cells HEK 293 cells used were Flp-In T-REX 293 cells corresponding to a Tet-On inducible system. The cells were cultured at 37 ° C. in a CO ₂ incubator.

(3) Transfer of R-CEPIA1er (described in Non-patent Document 13) to HEK293 cells and disease mutant RyR gene The disease mutant RyR gene is transferred to Flp-In T-REX 293 cells by lipofection to obtain hygromycin. A stable expression strain was established by culturing in the added medium for 10 to 14 days. The R-CEPIA1er expression vector is introduced into this disease mutant RyR stable expression strain by the lipofection method, and cultured in a medium to which G418 is added to obtain a stable expression strain which double-expresses the disease mutant RyR gene and R-CEPIA1er. Established.

(4) Transfer of R-CEPIA1er and wild-type RyR gene into HEK293 cells The wild-type RyR gene is introduced into Flp-In T-REX 293 cells by lipofection method and cultured in a medium supplemented with hygromycin for 10 to 14 days Stable expression strain was established. R-CEPIA1er expression vector is introduced into this disease mutant RyR stable expression strain by the lipofection method, and cultured in a medium to which G418 is added to establish a stable expression strain which dually expresses the wild-type RyR gene and R-CEPIA1er did.

(5) Measurement of fluorescent endoplasmic reticulum Ca ²⁺ concentration Cultured cells expressing the disease-altered RyR and R-CEPIA1er, and cultured cells expressing the wild-type RyR and R-CEPIA1er are respectively seeded in a 96-well plate and 37 Culturing was carried out for 24 hours. The medium was changed to a medium to which doxycycline was added, and culture was further continued for 24 hours to induce RyR expression. The medium was changed to Krebs solution before measurement. Fluorescence measurements were performed on a FlexStation. R-CEPIAl was excited at 560 nm to obtain fluorescence at 610 nm. The 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 to measure the concentration of Ca ²⁺ concentration in fluorescent vesicles.

(6) Test compounds The compounds subjected to the test are as follows.
Compounds according to the present invention Compound 1: 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 (oxophosphoric acid), and compound 29: 5-octyl-8-oxo- [1,1 3] Dioxoro [4,5-g] quinoline-7-carboxylic acid

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

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

(7) Results The results are shown in FIGS.
FIG. 1 is a comparison of the RyR1 inhibitory activity of oxophosphoric acid (compound 20) with the reference compound DOCA. As compared to DOCA, oxophosphoric acid has improved RyR1 inhibitory activity.
FIG. 2 shows that RyR1 inhibition was improved by substituting an alkyl group at the nitrogen atom of DOCA as a control compound.
FIG. 3 shows the activity of a compound having a substituent at

position

6 or 7 with 1,4-dihydro-1-octyl-4-oxoquinoline-3-carboxylic acid (compound 8, N-C ₈ ) as a reference compound. Shows that RyR1 inhibitory activity was improved particularly in compounds having a substituent at the 7-position.
FIG. 4 shows that compound 29 has excellent RyR1 inhibitory activity against dandrolen, which is known as a therapeutic agent for malignant hyperthermia.

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

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

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

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

Pharmacological Test 3 Animal Test The compound of the present invention was tested for its therapeutic effect on RyR1 malignant hyperthermia mutation-introduced mice by the following method. The test method was performed by measuring the temperature rise of the mouse produced by isoflurane anesthesia.

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

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

(3) Isoflurane Anesthetic Test Wild-type or mutant mice were anesthetized with 2% isoflurane and then maintained with 1% isoflurane. The mouse rectal temperature was measured continuously with a digital thermometer (Tateyama Scientific Industry, thermistor high accuracy 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. Wild-type mice (○) gradually increased in rectal temperature over 30 minutes after isoflurane anesthesia but did not die. The mutant mice (●) had a sudden rise in rectal temperature 15 minutes after anesthesia, and all muscle contraction occurred from 26 to 33 minutes and all died (+ marks in the figure). On the other hand, mutant mice pre-administered with Compound 29 (29) suppressed temperature rise and did not die. This result shows a remarkable preventive effect of compound (X) on malignant hyperthermia model mice.

Claims

General formula (I)

[Wherein, 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 have a C3-12 carbon ring, or a 4 to 12-membered complex] The ring may be substituted,
X represents CR 4 or N,
R 2 , R 3 and R 4 are each independently a hydrogen atom, a C1-6 alkyl group, a C2-6 alkenyl group, a C2-6 alkynyl group, a C1 to 3 fluoroalkyl group, a C1 to 6 alkoxy group, OR 10 (wherein, R 10 represents a hydrogen atom or a phenyl group), NR 11 R 12 (wherein, R 11 and R 12 each independently represent a hydrogen atom, a C1-6 alkyl group or a phenyl group) ), COR 13 (wherein, R 13 is a hydroxyl group, OR 20 (wherein, R 20 is a hydrogen atom, a C1-6 alkyl group or a phenyl group)) or NR 21 R 22 (wherein, R 21 and R 22 each independently represents a hydrogen atom, a C 1 to 6 alkyl group or a phenyl group)), a cyano group, a nitro group, a halogen atom, a C 1 to 3 fluorinated alkoxy group, a C 1 to 6 alkyl Thio, C1 ~ 6 alkylthio group, C3 ~ 12 shows a carbocyclic or 4-12 membered heterocyclic ring, R 2 and R 3 represents a since by C3 ~ 5 alkylene group or C1 ~ 3 alkylenedioxy group together Well,
Y is COR 30 (R 30 is a hydroxyl group, OR 40 (wherein R 40 is a C1-6 alkyl group or a phenyl group)) or NR 41 R 42 (wherein R 41 and R 42 are each independently. , A hydrogen atom, a C.sub.1-6 alkyl group or a phenyl group is shown). ]
A therapeutic or preventive agent for a ryanodine receptor related disease, which comprises the compound represented by or the pharmaceutically acceptable salt thereof as an active ingredient.
The compound represented by the general formula (I), wherein X is CH, R 1 is a C 4-10 alkyl group, R 2 is a hydrogen atom, and R 3 is a C 1-4 alkyl group. The therapeutic or prophylactic agent according to 1).
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 comprising a 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)

[Wherein, 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, and R 4A represents a hydrogen atom, a C 1-4 alkyl group or a C 1-4 alkoxy group,
One of R 2A and R 3A is a hydrogen atom, a C 1-4 alkyl group or a C 1-4 alkoxy group, and the other is a hydrogen atom, a C 1-4 alkyl group, a C 1-4 alkoxy group, a cyano group, a halogen atom, trihalomethyl Group, a trihalomethyloxy group or a methylthio group, or R 2A and R 3A together represent a C 3-5 alkylene group or an -O-CH 2 -O- group (however, (i) R 2A and When R 3A is both hydrogen atoms, R 1A represents a cyclohexylmethyl group,
(Ii) when R 1A is n-propyl, R 2A is not methyl or methoxy;
(Iii) When R 2A and R 3A are taken together to represent an —O—CH 2 —O— group, R 1A is not an octyl 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 C 1-4 alkyl group or a C 1-4 alkoxy group, and the other is a C 1-4 alkyl group, a C 1-4 alkoxy group, a halogen atom, a cyano group or a trihalomethyl group Or R 2 and R 3 taken together represent an —O—CH 2 —O— group (wherein (i) when R 1A is n-propyl, R 2A is not a methyl or methoxy group,
(Ii) R 1A is not an octyl group when R 2A and R 3A together represent an -O-CH 2 -O- group)]
A 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] dioxoro [4,5-g] quinoline-7-carboxylic acid,
(11) 7-Methoxy-1-propyl-1,4-dihydro-4-oxo-1,8-naphthyridine-3-carboxylic acid,
(12) 1,4-dihydro-7-methoxy-1-octyl-4-oxo-1,8-naphthyridine-3-carboxylic acid,
(13) 1-decyl-1,4-dihydro-7-methoxy-4-oxo-1,8-naphthyridine-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, which is selected from the group consisting of quinoline-3-carboxylic acid or a pharmaceutically acceptable salt thereof.