JP2018168083A - Novel quinolinecarboxylic acid derivative and medicament containing the same - Google Patents

Abstract

To provide a compound that inhibits the formation of amyloid fibrils, an amyloid fibrils formation inhibitor containing the compound, and therapeutic or preventive agents for neurodegenerative diseases.SOLUTION: The present invention provides a compound represented by formula (I). [X is -(CH)-; n is an integer of 0-3; Y is a single bond or a divalent linking group such as an alkyl group; Ar is a substituted/unsubstituted phenyl group or pyridyl group; Ris a substituted/unsubstituted alkyl group; Ar and R, Ar and Y or Rand Y may form a ring together; R-Rindependently represent H, a Calkyl group or the like].SELECTED DRAWING: None

Description

  The present invention relates to an amyloid fibril formation inhibitor. The present invention particularly relates to a therapeutic or prophylactic agent for neurodegenerative diseases by inhibiting amyloid fibril formation.

  In Japan, the population is aging at an unprecedented rate, and the number of patients with dementia centered on Alzheimer's disease is increasing. At present, the number of patients with dementia in Japan is estimated to be about 4.5 million, and it is estimated that the number of patients will increase with aging in the future. Since care for these patients with dementia is an economic burden, it is urgent to establish an effective treatment as soon as possible.

  Pathological features commonly seen in Alzheimer's disease patients include: (1) neuronal loss, (2) formation of patchy amyloid deposits (senile plaques), and (3) fibrous mass in neurons. There are three accumulations of (neurofibrillary tangles) that cause cognitive impairment (Alzheimer-type dementia), a clinical manifestation of Alzheimer's disease. As treatment for Alzheimer-type dementia, symptomatic treatment for temporarily improving symptoms of dementia is performed using a cholinesterase inhibitor, an NMDA inhibitor or the like. However, the effect is extremely limited, and establishment of a fundamental therapy that suppresses the onset and progression of Alzheimer's disease is strongly desired.

  In order to establish the basic therapy for Alzheimer's disease, it is necessary to elucidate the pathogenesis of Alzheimer's disease. Alzheimer's disease is known to be familial (hereditary) Alzheimer's disease caused by genetic factors and sporadic Alzheimer's disease without genetic background. And risk factors are being clarified. Since familial Alzheimer's disease and sporadic Alzheimer's disease share the same pathological findings and clinical symptoms, it is predicted that a common mechanism exists in the onset process. Research on the mechanism is expected to lead to the elucidation of the onset mechanism of sporadic Alzheimer's disease.

  One of genes causing familial Alzheimer's disease is a gene encoding an amyloid precursor protein (hereinafter referred to as APP). APP is cleaved by β-secretase and γ-secretase to produce amyloid β (hereinafter referred to as Aβ). Aβ has Aβ40 consisting of 40 amino acids and Aβ42 with 2 amino acids added to the C-terminus of Aβ40 due to differences in the cleavage point in APP, but Aβ40 is superior in physiological production. In contrast, senile plaques contain a large amount of Aβ42. Furthermore, it has been clarified that mutant APP increases the expression ratio of Aβ42 and that Aβ42 is more likely to aggregate than Aβ40. Based on these facts, it has been proposed that Aβ42 aggregates first, and Aβ40 aggregates and accumulates using it as a nucleus, so that amyloid fibril formation proceeds and Alzheimer's disease develops.

  Aggregation of Aβ in the brain is caused by the binding of GM1 ganglioside (hereinafter referred to as GM1 in the present specification) constituting the nerve cell membrane to a complex (GM1 binding type Aβ: hereinafter referred to as GAβ in this specification). It has been found that the process starts from the formation of (non-patent documents 1 and 2). In addition, GM1 aggregates in a nerve cell membrane depending on cholesterol concentration to form a cluster, and that Aβ specifically binds to this GM1 cluster to form GAβ. It has been confirmed by analysis using vesicles (Non-patent Document 3). Furthermore, analysis using an antibody that specifically recognizes GAβ has confirmed that GAβ has a structure different from that of soluble Aβ (Non-patent Document 4). From these findings, it has been proposed that when Aβ binds to the GM1 cluster, it changes to GAβ having a different structure, and this GAβ becomes a “seed” that promotes Aβ aggregation. Since then, many research results supporting this hypothesis have been obtained (Non-Patent Documents 5 to 9).

  Until now, various compounds have been developed and subjected to clinical trials for the purpose of developing fundamental treatment and prevention methods for Alzheimer's disease. Well-known ones include, for example, inhibitors for β-secretase that cleaves the N-terminal side of Aβ when Aβ is generated, and inhibitors for γ-secretase that cleaves the C-terminal side of Aβ (patent) Document 1, Non-Patent Document 10). These inhibitors were expected to be able to suppress amyloid fibril formation by inhibiting Aβ production. However, both β-secretase and γ-secretase are not enzymes using only APP as a specific substrate, and as a result, γ-secretase particularly inhibits Notch signals that are deeply involved in development and differentiation. The problem is that serious side effects are caused by inhibiting the physiological metabolism of APP itself. Many low molecular weight compounds that inhibit Aβ polymerization have also been developed (Patent Documents 2 to 6). These compounds inhibit the elongation of amyloid fibrils or disrupt the fibrils by inhibiting Aβ polymerization. However, it is suggested that if the administration of the compound is stopped, the polymerization of Aβ is resumed, and the fiber fragmentation itself may give rise to a new seed of polymerization, which is sufficient for the formation of amyloid fibrils. However, it has not been able to obtain a sufficient suppression effect.

  On the other hand, a method for suppressing amyloid fibril formation using an antibody (4396C antibody) that specifically binds to GAβ has been reported (Patent Document 7). The 4396C antibody specifically inhibits the formation of amyloid fibrils by specifically recognizing and binding to GAβ and inhibiting Aβ polymerization by GAβ. However, there is a problem that it takes a great amount of time and money to produce the antibody, and the administration method of the drug using the antibody is limited, and there is a problem of delivery to the brain. There is a problem that it is difficult to use.

US Patent Application Publication No. 201200233156 International Publication No. 2009/103652 International Publication No. 2003/045923 US Pat. No. 7,687,512 US Patent Application Publication No. 201000063077 US Patent Application Publication No. 20060223812 Japanese Patent No. 4102850

Yanagisawa, K .; et al. , Nat. Med. , Vol. 1, pp. 1062-1066 (1995) Yanagisawa, K .; et al. , Neurobiol. Aging, Vol. 19, pp. S65-67 (1998) Kakio, A .; et al. , J. et al. Biol. Chem. , Vol. 276, pp. 24985-24990 (2001) Yanagisawa, K .; et al. , FEBS Lett. , Vol. 420, pp. 43-46 (1997) McLaurin, J. et al. , And Chakrabarty, A .; , J. et al. Biol. Chem. , Vol. 271, pp. 26482-26489 (1996) Choo-Smith, L.M. P. , And Surewicz, W .; K. , FEBS Lett. , Vol. 402, pp. 95-98 (1997) Choo-Smith, L.M. P. et al. , J. et al. Biol. Chem. , Vol. 272, pp. 22987-22990 (1997) Matsuzaki, K .; et al. , Biochemistry, Vol. 38, pp. 4137-4142 (1999) Koppaka, V.K. and Axelsen, P.A. H. , Biochemistry, Vol. 39, pp. 10011-10016 (2000) Shuto, D.C. et al. Bioorg. Med. Chem. Lett. , Vol. 13, pp. 4273-4276 (2003)

  The present invention has been made for the purpose of solving various problems of the prior art and providing a fundamental prevention / treatment method for neurodegenerative diseases such as Alzheimer's disease. Specifically, the polymerization of Aβ is specifically performed. An object of the present invention is to provide an amyloid fibril formation inhibitor that can be inhibited and is excellent in clinical application.

  As a result of intensive studies, the present inventors succeeded in obtaining a low molecular weight compound that inhibits Aβ polymerization by specifically binding to GAβ.

［式中、
Ｘは、−（ＣＨ_２）_ｎ−
（式中、ｎは、０〜３の整数である）
であり、
Ｙは、単結合、または、主鎖が炭素原子、窒素原子および酸素原子からなる群から選択される１種または２種以上の原子から構成される、原子数が１〜４である２価の連結基であり、
Ａｒは、置換もしくは非置換のフェニル基またはピリジル基であり、
Ｒ_１は、置換もしくは非置換のＣ_１−６アルキル基または置換もしくは非置換のＣ_３−７シクロアルキル基であり、
ここで、ＡｒおよびＲ_１、ＡｒおよびＹ、またはＲ_１およびＹは、共同して環を形成してもよく、
Ｒ_２、Ｒ_３およびＲ_４は、それぞれ独立して、水素原子、Ｃ_１−２０アルキル基、Ｃ_１−２０ヒドロキシアルキル基、Ｃ_１−２０アルコキシ基またはハロゲン原子である］
で表される化合物もしくはその薬学的に許容される塩またはそれらの溶媒和物を含んでなる、アミロイド線維形成抑制剤を提供するものである。 That is, the present invention, according to one embodiment, is represented by the general formula (I):

[Where:
X represents — (CH ₂ ) _n —.
(In the formula, n is an integer of 0 to 3)
And
Y is a single bond or a divalent one having 1 to 4 atoms, the main chain being composed of one or more atoms selected from the group consisting of carbon atoms, nitrogen atoms and oxygen atoms. A linking group,
Ar is a substituted or unsubstituted phenyl group or pyridyl group;
R ₁ is a substituted or unsubstituted C _1-6 alkyl group or a substituted or unsubstituted C _3-7 cycloalkyl group,
Here, Ar and R ₁ , Ar and Y, or R ₁ and Y may jointly form a ring,
R ₂ , R ₃ and R ₄ are each independently a hydrogen atom, a C _1-20 alkyl group, a C _1-20 hydroxyalkyl group, a C _1-20 alkoxy group or a halogen atom]
Or a pharmaceutically acceptable salt thereof, or a solvate thereof, to provide an amyloid fibril formation inhibitor.

［式中、
Ｙは、単結合、または、置換もしくは非置換のＣ_１−２０アルキル基、アミド基、アミン基、エーテル基および置換もしくは非置換のＣ_３−２０シクロアルキル基からなる群から選択される２価の連結基であり、
Ａｒは、置換もしくは非置換のフェニル基であり、
Ｒ_１は、置換もしくは非置換のＣ_１−４アルキル基または置換もしくは非置換のＣ_３−６シクロアルキル基であり、
ここで、ＡｒおよびＲ_１、ＡｒおよびＹ、またはＲ_１およびＹは、共同して環を形成してもよく、
Ｒ_２およびＲ_３は、それぞれ独立して、水素原子、Ｃ_１−２０アルキル基またはハロゲン原子である］
で表されるものであることが好ましい。 The compound represented by the general formula (I) is represented by the general formula (Ia):

[Where:
Y is a single bond or a divalent group selected from the group consisting of a substituted or unsubstituted C _1-20 alkyl group, an amide group, an amine group, an ether group, and a substituted or unsubstituted C _3-20 cycloalkyl group. A linking group of
Ar is a substituted or unsubstituted phenyl group,
R ₁ is a substituted or unsubstituted C _1-4 alkyl group or a substituted or unsubstituted C _3-6 cycloalkyl group;
Here, Ar and R ₁ , Ar and Y, or R ₁ and Y may jointly form a ring,
R ₂ and R ₃ are each independently a hydrogen atom, a C _1-20 alkyl group or a halogen atom]
It is preferable that it is represented by these.

  In addition, according to one embodiment, the present invention provides one or more compounds selected from the group consisting of compounds Nos. 1 to 29 described in Table 1A and Table 1B, or pharmaceutically acceptable salts thereof, or An amyloid fibril formation inhibitor comprising the solvate of

  The compound is preferably selected from the group consisting of compounds Nos. 1, 2, 4, 7, 9, 10, 13, 15, 18, 21, 23-25, 27 and 28.

  Alternatively, the compound is preferably selected from the group consisting of compounds Nos. 1, 2, 7, 9, 23 and 25.

  Moreover, according to one embodiment of the present invention, there is provided a therapeutic or prophylactic agent for neurodegenerative diseases containing the amyloid fibril formation inhibitor.

  The neurodegenerative disease is preferably Alzheimer's disease.

  Moreover, according to one Embodiment, this invention provides the compound selected from the group which consists of compound numbers 1-29, its pharmaceutically acceptable salt, or those solvates.

  The compound is preferably selected from the group consisting of compounds Nos. 2-12 and 14-29.

  The amyloid fibril formation inhibitor according to the present invention, like the existing anti-GAβ antibody (4396C antibody), specifically binds to GAβ and blocks the initiation of polymerization of amyloid fibrils, thereby reliably forming amyloid fibrils. Can be suppressed. Moreover, since the active ingredient of the amyloid fibril formation inhibitor according to the present invention is a low-molecular compound, (1) it can be easily synthesized and can be prepared in large quantities at low cost, and (2) it does not exhibit antigenicity. Therefore, it is superior to antibodies in that it is highly safe, (3) has high molecular stability, and can secure a stable GAβ binding ability in various dosage forms.

  Further, by using the amyloid fibril formation inhibitor according to the present invention, it is possible to provide a fundamental prevention / treatment method for neurodegenerative diseases such as Alzheimer's disease.

  Hereinafter, the present invention will be described in detail, but the present invention is not limited to the embodiments described in the present specification.

で表される化合物（以降、本明細書では、「化合物（Ｉ）」と記載する）もしくはその薬学的に許容される塩またはそれらの溶媒和物を含んでなる、アミロイド線維形成抑制剤である。 The present invention, according to a first embodiment, has the general formula (I):

Or a pharmaceutically acceptable salt thereof or a solvate thereof, or an amyloid fibril formation inhibitor comprising the compound represented by formula (hereinafter referred to as “compound (I)”) or a pharmaceutically acceptable salt thereof: .

  “Amyloid fibrils” are insoluble substances formed by aggregation of Aβ in a fibrous form. Amyloid fibrils are formed by first binding Aβ to GM1 constituting the nerve cell membrane to form a complex GAβ, and then using Aβ as a “seed” to aggregate Aβ. The amyloid fibril formation inhibitor according to this embodiment suppresses the formation of amyloid fibrils by inhibiting the aggregation of Aβ with respect to GAβ.

  “Aβ” is a peptide produced by cleaving APP with β-secretase and γ-secretase, and includes Aβ40, Aβ42, etc., which differ in the number of amino acids due to differences in cleavage points in APP.

  “APP” includes RefSeq accession numbers NP_001129601.1, NP_001129602.1, NP_0011191232.2, NP_9588177.1 in the NCBI RefSeq database (https://www.ncbi.nlm.nih.gov/RefSeq/). In addition to human APP consisting of the amino acid sequences registered as NP_0011921231.1, NP_958816.1, NP_0011921230.1, NP_0004755.1, NP_001129488.1, NP_0011299603.1, it is cleaved by β-secretase and γ-secretase to produce Aβ. 80% or more, preferably 90% or more, and more preferably about 95% with the above amino acid sequence. Protein comprising the amino acid sequence identity with the above may be included. Amino acid sequence identity can be calculated using sequence analysis software or using programs routine in the art (FASTA, BLAST, etc.).

  In addition, “APP” includes substitution, deletion, insertion and / or substitution of one to several amino acids in the above amino acid sequence, as long as it is cleaved by β secretase and γ secretase to produce Aβ. Proteins consisting of added amino acid sequences can be included. Here, “1 to several” is, for example, “1 to 30”, preferably “1 to 10”, and particularly preferably “1 to 5”.

  The amyloid fibril formation inhibitor of this embodiment comprises Compound (I) or a pharmaceutically acceptable salt thereof or a solvate thereof.

In compound (I), X is — (CH ₂ ) _n —. In said formula, n is an integer of 0-3, Preferably it is 0.

In the compound (I), Y has a single bond, or one or more atoms selected from the group consisting of a carbon atom, a nitrogen atom and an oxygen atom in the main chain, and the number of atoms is 1 to 2 4 is a divalent linking group. The side chain of the said coupling group is not specifically limited, For example, you may have a substituent as described below. Preferably, in compound (I), Y is a single bond or a substituted or unsubstituted C _1-20 alkyl group, an amide group, an amine group, an ether group, and a substituted or unsubstituted C _3-10 cycloalkyl group. A divalent linking group selected from the group consisting of

The C _1-20 alkyl group is preferably C _1-10 , particularly preferably C _1-6 . The alkyl group includes both linear and branched chain forms. The C _3-20 cycloalkyl group is preferably C _3-10 , particularly preferably C _3-6 .

In this embodiment, the alkyl group and cycloalkyl group may not be substituted, and one or more hydrogen atoms may be substituted with a substituent. The substituents in this case are C _1-20 alkyl group, C _1-20 haloalkyl group, C _1-20 alkoxy group, C _1-20 haloalkoxy group, C _1-20 hydroxyalkoxy group, C _1-20 alkoxyalkyl. Group, C _1-20 haloalkoxyalkyl group, C _1-20 alkoxyalkoxy group, C _1-20 haloalkoxyalkoxy group, C _1-20 hydroxyalkyl group, C _1-20 aminoalkyl group, C _1-20 alkylamino Alkyl group, C _1-20 dialkylaminoalkyl group, C _1-20 alkylamino group, C _1-20 alkylaminocarbonyl group, C _1-20 dialkylaminocarbonyl group, C _1-20 alkoxycarbonyl group, C _1-20 alkoxycarbonylalkyl _{group, C 1-20} acyl _{groups, C 1-20} acyl Alkyl _{group, C 1-20} alkylthio _{group, C 1-20} alkylenedioxy _{group, C 1-20} halo alkylenedioxy group, a halogen atom, an amino group, a nitro group, aminosulfonyl _{group, C 1-20} alkylaminosulfonyl group , A C _1-20 alkylsulfonylamino group, an aryl group, a heteroaryl group, a cyano group, a thiol group, and a hydroxyl group. The number of substituents and the substitution position are not particularly limited, but the number of substituents is preferably 0 to 3.

  In compound (I), Ar is a substituted or unsubstituted phenyl group or pyridyl group. In the present embodiment, the phenyl group may not be substituted, and one or more hydrogen atoms may be substituted with a substituent. The substituents in this case are the same as those defined above.

In compound (I), R ₁ is a substituted or unsubstituted C _1-6 alkyl group or a substituted or unsubstituted C _3-7 cycloalkyl group, preferably a substituted or unsubstituted C _1-4 alkyl group. Or a substituted or unsubstituted C _3-6 cycloalkyl group. In this embodiment, the alkyl group and cycloalkyl group may not be substituted, and one or more hydrogen atoms may be substituted with a substituent. The substituents in this case are the same as those defined above.

In compound (I), Ar and R ₁ , Ar and Y, or R ₁ and Y may jointly form a ring. The ring may be a single ring or a multi-ring (for example, a bicyclo ring).

In compound (I), R ₂ , R ₃ and R ₄ are each independently a hydrogen atom, a C _1-20 alkyl group, a C _1-20 hydroxyalkyl group, a C _1-20 alkoxy group or a halogen atom. Preferably a hydrogen atom, a C _1-20 alkyl group or a halogen atom. Particularly preferably, R ₄ is a hydrogen atom.

The C _1-20 alkyl group, C _1-20 hydroxyalkyl group, and C _1-20 alkoxy group are preferably C _1-10 , particularly preferably C _1-6 . In addition, the alkyl moiety in the above group includes both linear and branched forms.

一般式（Ｉａ）中、Ｙは、単結合、または、置換もしくは非置換のＣ_１−２０アルキル基、アミド基、アミン基、エーテル基および置換もしくは非置換のＣ_３−２０シクロアルキル基からなる群から選択される２価の連結基であり、Ａｒは、置換もしくは非置換のフェニル基であり、Ｒ_１は、置換もしくは非置換のＣ_１−４アルキル基または置換もしくは非置換のＣ_３−６シクロアルキル基であり、ここで、ＡｒおよびＲ_１、ＡｒおよびＹ、またはＲ_１およびＹは、共同して環を形成してもよく、Ｒ_２およびＲ_３は、それぞれ独立して、水素原子、Ｃ_１−２０アルキル基またはハロゲン原子であることが好ましい。 The compound (I) is preferably one represented by the following general formula (Ia).

In general formula (Ia), Y is a single bond or a substituted or unsubstituted C _1-20 alkyl group, an amide group, an amine group, an ether group, and a substituted or unsubstituted C _3-20 cycloalkyl group. A divalent linking group selected from the group, Ar is a substituted or unsubstituted phenyl group, R ₁ is a substituted or unsubstituted C _1-4 alkyl group or a substituted or unsubstituted C _{3- 6} cycloalkyl groups, wherein Ar and R ₁ , Ar and Y, or R ₁ and Y may form a ring together, R ₂ and R ₃ are each independently hydrogen It is preferably an atom, a C _1-20 alkyl group or a halogen atom.

  Unless otherwise specified, the compound (I) used in the present embodiment also includes stereoisomers such as tautomers, geometric isomers (for example, E isomer, Z isomer, etc.), enantiomers and the like. .

  Compound (I) used in the present embodiment can be synthesized by appropriately combining various conventionally known methods with the chemical synthesis methods described in the following examples and chemical synthesis methods corresponding thereto.

  Alternatively, when compound (I) used in the present embodiment is selected from commercially available compounds, commercially available compounds may be purchased. Commercially available compounds are, for example, domestically, through Namiki Shoji Co., Ltd. or Kishida Chemical Co., Ltd. , ChemDiv etc. can be purchased.

  The amyloid fibril formation inhibitor of the present embodiment includes not only those comprising the above compound (I) but also those comprising a pharmaceutically acceptable salt of the above compound (I) or a solvate thereof. Is done.

  In the present embodiment, “pharmaceutically acceptable” means not harmful when a drug is used. The pharmaceutically acceptable salt in the present embodiment is, for example, an alkali metal salt such as a lithium salt, a sodium salt, a potassium salt, a magnesium salt, a calcium salt or the like when an acidic group is present in the compound (I) Alkaline earth metal salts; ammonia, methylamine, dimethylamine, trimethylamine, dicyclohexylamine, tris (hydroxymethyl) aminomethane, N, N-bis (hydroxyethyl) piperazine, 2-amino-2-methyl-1-propanol, Examples include amine addition salts such as ethanolamine, N-methylglucamine, and L-glucamine; or basic amino acid addition salts such as lysine, δ-hydroxylysine, and arginine. Further, for example, when a basic group is present in the compound (I), inorganic acid salts such as hydrochloride, hydrobromide, nitrate, sulfate, phosphate, etc .; methanesulfonic acid, benzenesulfonic acid, P-Toluenesulfonic acid, acetic acid, propionate, tartaric acid, fumaric acid, maleic acid, malic acid, oxalic acid, succinic acid, citric acid, benzoic acid, mandelic acid, cinnamic acid, lactic acid, glycolic acid, glucuronic acid, ascorbine Examples thereof include organic acid salts such as acid, nicotinic acid and salicylic acid; and acidic amino acid addition salts such as aspartic acid and glutamic acid.

  The pharmaceutically acceptable “solvate” in the present embodiment includes, for example, hydrate, alcoholate, dimethylformamide, dimethylacetamide, N-methyl-2-pyrrolidone, chloroform, dichloroethane, dichloromethane, Diethyl ether, methyl t-butyl ether, ethylene glycol monomethyl ether, ethylene glycol dimethyl ether, ethylene glycol monoethyl ether, ethylene glycol diethyl ether, diisopropyl ether, tetrahydrofuran, dioxane, acetone, methyl ethyl ketone, isobutyl methyl ketone, toluene, benzene, o-xylene , M-xylene, p-xylene, ethyl acetate, propyl acetate, butyl acetate, propylene carbonate, diethyl carbonate, dimethyl carbonate, hexane, Tan, heptane, etc. solvates with pharmaceutically acceptable solvents selected from the group consisting of octane and cyclohexane and the like, preferably hydrates.

  The compound represented by compound (I) or a pharmaceutically acceptable salt thereof or a solvate thereof may be used alone or a mixture of two or more thereof as an active ingredient of an amyloid fibril formation inhibitor. it can. The amyloid fibril formation inhibitor of the present embodiment may be composed of only an active ingredient, but generally, as an optional ingredient, a known pharmaceutically acceptable diluent, carrier, excipient, etc. May be included.

  In order to produce the amyloid fibril formation inhibitor of the present embodiment, the compound represented by Compound (I) or a pharmaceutically acceptable salt thereof or a solvate thereof, if necessary, is publicly known according to a conventional method. What is necessary is just to formulate in combination with these carriers. In the amyloid fibril formation inhibitor, the compound represented by Compound (I) or a pharmaceutically acceptable salt thereof, or a solvate thereof has an appropriate intake amount in a range corresponding to each form as an active ingredient. So long as it is contained. In the amyloid fibril formation inhibitor, the compound represented by Compound (I) or a pharmaceutically acceptable salt thereof or a solvate thereof is usually administered at a dose of 0.001 mg / kg (body weight) per day for an adult. As described above, the content in the drug is preferably determined so as to be preferably 0.01 mg / kg (body weight) or more. However, the content is not limited to this range, and may be appropriately determined depending on the patient's symptoms, age, sex, and the like. It can be adjusted. The upper limit of the dose is preferably 100 mg / kg (body weight) or less, more preferably 10 mg / kg (body weight) or less per day.

  The amyloid fibril formation inhibitor of the present embodiment can be formulated into various dosage forms. Examples of the dosage form include tablets, capsules, granules, powders, syrups, suspensions, suppositories, Examples include ointments, creams, gels, patches, inhalants, injections and the like. Therefore, the amyloid fibril formation inhibitor of this embodiment can be administered by various methods such as oral administration, intraperitoneal administration, intradermal administration, intravenous administration, intramuscular administration, and intracerebral administration.

  When the amyloid fibril formation inhibitor is used as an oral preparation, it can be made into solid preparations such as tablets, capsules, powders and granules. In this case, suitable additives such as additives such as starch, lactose, sucrose, mannitol, carboxymethylcellulose, corn starch, and inorganic salts, and if desired, binders, disintegrants, lubricants, coloring agents, fragrances, etc. Can be blended. When the solid preparation is a tablet or pill, it may be coated with a sugar coating such as sucrose, gelatin or hydroxypropylcellulose, or a film of a gastric or enteric substance, if desired. Alternatively, the oral preparation of an amyloid fibril formation inhibitor can be a liquid such as a syrup, and in this case, sterile water, physiological saline, ethanol, or the like can be used as a carrier. If desired, auxiliary agents such as suspending agents, sweetening agents, flavoring agents, preservatives and the like may be added.

  When the amyloid fibril formation inhibitor is used as a parenteral preparation, it can be a liquid such as an injection or a rectal agent. In this case, the active ingredient is dissolved in a diluent such as distilled water for injection, physiological saline, aqueous glucose solution, vegetable oil for injection, sesame oil, peanut oil, soybean oil, corn oil, propylene glycol, polyethylene glycol, etc. by conventional methods. Alternatively, it can be prepared by suspending and adding a disinfectant, a stabilizer, an isotonic agent, a soothing agent, or the like, if necessary. In addition, a solid composition can be produced and dissolved in sterile water or a sterile solvent for injection before use.

  Moreover, the parenteral preparation of an amyloid fibril formation inhibitor can be, for example, a sustained release preparation such as a microcapsule, and can also be directly administered into the brain. Sustained-release preparations can be prepared using a carrier that can prevent immediate release from the body. As a preferable carrier, for example, a biodegradable / biocompatible polymer such as ethylene vinyl acetate, polyanhydride, polyglycolic acid, collagen, polyorthoester, and polylactic acid can be used. Liposomes can also be used as the carrier. Preferred liposomes may be, but are not limited to, those that have been purified and prepared by the reverse phase evaporation method using a lipid composition comprising phosphatidylcholine, cholesterol and PEG-derivatized phosphatidylethanolamine (PEG-PE).

  Furthermore, the amyloid fibril formation inhibitor of this embodiment may contain a pharmaceutically acceptable additive such as a coloring agent, a preservative, a fragrance, a flavoring agent, and a sweetening agent and other therapeutic agents as desired. it can.

  Moreover, it is preferable to mix | blend a stabilizer when formulating an amyloid fibril formation inhibitor. Examples of the stabilizer include albumin, globulin, gelatin, mannitol, glucose, dextran, ethylene glycol and the like.

  The amount of the active ingredient contained in the preparation varies depending on various conditions, for example, the type of extraction solvent, the amount of solvent used, and the like. Therefore, the amount of the active ingredient may be sufficient in an amount smaller than the above preferable intake amount or may be necessary beyond the range.

  The amyloid fibril formation inhibitor of this embodiment can fundamentally and reliably suppress the formation of amyloid fibrils. Therefore, it is useful for uses such as treatment and prevention of diseases associated with amyloid fibril formation.

  According to a second embodiment, the present invention comprises one or more compounds selected from the group consisting of compounds of compound numbers 1 to 29, or a pharmaceutically acceptable salt thereof, or a solvate thereof. , An amyloid fibril formation inhibitor. The compounds of compound numbers 1 to 29 shown below can inhibit the polymerization of Aβ with respect to GAβ and suppress the formation of amyloid fibrils.

[1] 4- [Benzyl (methyl) amino] -7-chloro-8-methylquinoline-3-carboxylic acid (Compound No. 1),
[2] 2- (7-chloro-4-{[(4-fluorophenyl) methyl] (methyl) amino} -8-methylquinolin-3-yl) acetic acid (Compound No. 2),
[3] 4- (Benzylamino) -7-chloro-8-methylquinoline-3-carboxylic acid (Compound No. 3),
[4] 7-chloro-8-methyl-4- [methyl (2-phenylethyl) amino] quinoline-3-carboxylic acid (Compound No. 4),
[5] 7-chloro-8-methyl-4- {methyl [(pyridin-3-yl) methyl] amino} quinoline-3-carboxylic acid (Compound No. 5),
[6] 7-chloro-8-methyl-4- {methyl [(pyridin-2-yl) methyl] amino} quinoline-3-carboxylic acid (Compound No. 6),
[7] 7-chloro-4-{[(4-fluorophenyl) methyl] (methyl) amino} -8-methylquinoline-3-carboxylic acid (Compound No. 7),
[8] 7-chloro-8-methyl-4-[(1-phenylethyl) amino] quinoline-3-carboxylic acid (Compound No. 8),
[9] 7-chloro-8-methyl-4- (phenylamino) quinoline-3-carboxylic acid (Compound No. 9),
[10] 7-chloro-8-methyl-4- [methyl (phenyl) amino] quinoline-3-carboxylic acid (Compound No. 10),
[11] 4- [Benzyl (methyl) amino] -7-chloroquinoline-3-carboxylic acid (Compound No. 11),
[12] 4- [Benzyl (methyl) amino] -8-methylquinoline-3-carboxylic acid (Compound No. 12),
[13] 7-chloro-8-methyl-4- (1,2,3,4-tetrahydroisoquinolin-2-yl) quinoline-3-carboxylic acid hydrochloride (Compound No. 13),
[14] 7-chloro-8-methyl-4-[(1,2,3,4-tetrahydronaphthalen-1-yl) amino] quinoline-3-carboxylic acid hydrochloride (Compound No. 14),
[15] 7-chloro-4-{[(4-chlorophenyl) methyl] (methyl) amino} -8-methylquinoline-3-carboxylic acid (Compound No. 15),
[16] 7-chloro-4-{[1- (4-chlorophenyl) ethyl] amino} -8-methylquinoline-3-carboxylic acid (Compound No. 16),
[17] 7-chloro-8-methyl-4- (3-phenylazetidin-1-yl) quinoline-3-carboxylic acid (Compound No. 17),
[18] 7-chloro-4-[(2,3-dihydro-1H-inden-1-yl) amino] -8-methylquinoline-3-carboxylic acid (Compound No. 18),
[19] 7-chloro-4- (4-chlorobenzamide) -8-methylquinoline-3-carboxylic acid (Compound No. 19),
[20] 4- [Benzyl (methyl) amino] -7-fluoro-8-methylquinoline-3-carboxylic acid (Compound No. 20),
[21] 7-chloro-4-{[1- (4-chlorophenyl) ethyl] (methyl) amino} -8-methylquinoline-3-carboxylic acid (Compound No. 21),
[22] 7-chloro-4- (2,3-dihydro-1H-isoindol-2-yl) -8-methylquinoline-3-carboxylic acid (Compound No. 22),
[23] 7-chloro-4-[(2,3-dihydro-1H-inden-1-yl) (methyl) amino] -8-methylquinoline-3-carboxylic acid (Compound No. 23),
[24] 2- {4- [Benzyl (methyl) amino] -7-chloro-8-methylquinolin-3-yl} acetic acid (Compound No. 24),
[25] 7-chloro-4- [4- (4-fluorophenyl) piperazin-1-yl] -8-methylquinoline-3-carboxylic acid (Compound No. 25),
[26] 7-chloro-4- [4- (4-fluorophenyl) -4-hydroxypiperidin-1-yl] -8-methylquinoline-3-carboxylic acid (Compound No. 26),
[27] 4- [Benzyl (2-hydroxyethyl) amino] -7-chloro-8-methylquinoline-3-carboxylic acid (Compound No. 27),
[28] 4- [Benzyl (propan-2-yl) amino] -7-chloro-8-methylquinoline-3-carboxylic acid (Compound No. 28), and [29] 7-Chloro-8-methyl-4- [(2S) -2-[(Phenylamino) methyl] pyrrolidin-1-yl] quinoline-3-carboxylic acid (Compound No. 29).

  More preferably, the compound used in the present embodiment is selected from the group consisting of compounds Nos. 1, 2, 4, 7, 9, 10, 13, 15, 18, 21, 23-25, 27 and 28. Is done. The above compounds have a strong inhibitory effect on amyloid fibril formation.

  The compound used in this embodiment is particularly preferably selected from the group consisting of compounds of compound numbers 1, 2, 7, 9, 23 and 25. The above compounds have a particularly strong inhibitory effect on amyloid fibril formation.

  As in the case of Compound (I) in the first embodiment, compounds of Compound Nos. 1 to 29 are appropriately combined with various conventionally known methods in the chemical synthesis methods described in the following Examples and the chemical synthesis methods equivalent thereto. Can be synthesized. Or when selecting from a commercially available compound, you may purchase a commercially available compound.

  The amyloid fibril formation inhibitor of this embodiment includes those comprising the above-mentioned specific compound, as well as those comprising a pharmaceutically acceptable salt of the above compound or a solvate thereof. The “pharmaceutically acceptable salt” and “solvate” in the present embodiment are the same as those defined in the first embodiment.

  The amyloid fibril formation inhibitor of this embodiment may be prepared in the same manner as the amyloid fibril formation inhibitor of the first embodiment, except that the specific compound is used as the compound (I) in the first embodiment. . Compounds 1, 2, 4 to 7, 10 to 13, 15, 17, and 20 to 29 are compounds having a structure represented by the general formula (I). Compounds 3, 8, 9, 14, 16, 18, and 19 are compounds having a structure similar to the structure represented by formula (I), and can be used in place of compound (I).

  According to the third embodiment, the present invention is a therapeutic or prophylactic agent for neurodegenerative diseases containing the amyloid fibril formation inhibitor. The therapeutic or prophylactic agent for neurodegenerative diseases according to this embodiment is a compound represented by compound (I) or a compound selected from compound numbers 1 to 29, or a pharmaceutically acceptable salt thereof, or a solvent thereof. An amyloid fibril formation inhibitor comprising a single substance or a mixture of two or more selected from Japanese and an optional component. The arbitrary component referred to here may be a normal pharmaceutical ingredient, and is prepared as various commonly used forms such as tablets, capsules, powders, granules, liquids and the like. In addition, the therapeutic agent or preventive agent for neurodegenerative diseases according to this embodiment may further contain other active ingredients for treating or preventing neurodegenerative diseases as optional components.

  In this embodiment, `` treatment '' means to prevent or alleviate the progression and worsening of the disease state in an animal suffering from a neurodegenerative disease, and not only completely cure the disease, It also includes alleviating the symptoms of the disease. In addition, “prevention” means to prevent the disease from occurring in an animal that may suffer from a neurodegenerative disease.

  In the present embodiment, “neurodegenerative disease” means all diseases associated with (or resulting from) neuronal tissue degeneration associated with Aβ aggregation and amyloid fibril formation. Although it does not specifically limit as a neurodegenerative disease, For example, Alzheimer's disease, Parkinson's disease, Down's syndrome, cerebral amyloid angiopathy, Huntington's disease etc. are mentioned.

  The neurodegenerative disease that is the target of the therapeutic or prophylactic agent of this embodiment is preferably Alzheimer's disease. “Alzheimer's disease” includes familial (hereditary) Alzheimer's disease as well as so-called Alzheimer's disease (spontaneous Alzheimer's disease).

  The animal that is the target of the therapeutic or prophylactic agent of the present embodiment is, for example, a mammal such as a mouse, rat, rabbit, dog, cow, pig, sheep, non-human primate, human, and preferably a human. .

  The therapeutic agent or preventive agent for neurodegenerative diseases according to this embodiment is useful for the fundamental treatment or prevention of neurodegenerative diseases associated with the formation of amyloid fibrils.

  According to a fourth embodiment, the present invention is a compound selected from the group consisting of compounds of compound numbers 1 to 29, or a pharmaceutically acceptable salt thereof, or a solvate thereof.

  The compound of this embodiment is preferably selected from the group consisting of the compounds of Compound Nos. 2-12 and 14-29.

  The compound of the present embodiment can be synthesized by appropriately combining various conventionally known methods with the chemical synthesis methods described in the following examples and the chemical synthesis methods corresponding thereto.

  The “pharmaceutically acceptable salt” and “solvate” in the present embodiment are the same as those defined in the first embodiment.

  The compound according to the present embodiment or a pharmaceutically acceptable salt thereof or a solvate thereof can inhibit the polymerization of Aβ with respect to GAβ and suppress the formation of amyloid fibrils. Therefore, it is useful for uses such as treatment and prevention of diseases associated with amyloid fibril formation.

  The following examples further illustrate the present invention. In addition, these do not limit this invention at all.

<1. Screening for compounds that inhibit amyloid fibril formation>
In order to find a compound that specifically recognizes GAβ and suppresses the formation of amyloid fibrils, virtual screening using a molecular docking simulation program DOCK4 (https://dock.compbio.ucsf.edu/) was performed. In conducting virtual screening, we first constructed a GAβ complex model by molecular dynamics simulation. In the GM1 molecule, the structure optimization based on the molecular orbital calculation was applied to the sugar chain region considered to be involved in the binding to Aβ, and a partial charge was assigned to each atom. The three-dimensional coordinate data and partial charge values of each atom of these optimized structures are incorporated into the ff99 parameter set of the empirical force field used in AMBER, and a molecule of 5 ns in a molecular system in which water molecules are arranged Kinetic calculations were performed. Molecular dynamics calculation was performed on the sugar chain region of GM1, and trajectory poses were sampled. Using the obtained trajectory pose, docking with the three-dimensional structure of Aβ40 (PDB ID: 1AML) registered in the Protein Data Bank was performed. As a result of docking, 2 million docking poses were obtained. Among them, a docking pose with the least structural distortion and the most stable energy is extracted and used as an atomic model of GAβ. Using this atomic model of GAβ as an initial structure, molecular dynamics calculation of 20 ns is performed, structure clustering is performed based on positional information of the main chain Cα atom of Aβ, and compounds are bonded from the obtained representative structures. Extracted promising things in the pocket.

  Drug-like filtering is used to exclude compounds that are not desirable as drugs from a commercially available compound library consisting of a total of about 9 million compounds provided by Namiki Shoji Co., Ltd. and Kishida Chemical Co., Ltd., and clustering is performed using fingerprints. By extracting representative structures from these groups, a commercial compound library consisting of about 500,000 structures was created. A similar structure search was performed on the commercially available compound library using the scaffold extracted above as a query. For fingerprints for structure search, fingerprints of MACCS, Lingo, Circular, Path, and Tree were used. Among the above five types of fingerprints, as a result of searching by MACCS fingerprint, 4,344 compounds with a similarity of 0.8 or more and 1,220 compounds with a score of 0.9 or more were hit. In other fingerprints, the compound did not hit. From the hit compounds by the MACCS fingerprint, compounds having an aromatic substituent similar to the side chain of Trp were selected, and 542 compounds were extracted. Subsequently, a plurality of conformations including geometric isomers and optical isomers were generated from the two-dimensional structure of each compound, converted into three-dimensional structure coordinates, and docked with GAβ. From each docking pose, compounds satisfying the pharmacophore for forming aromatic stacking with Phe19 were selected and narrowed down to 299. Further, the structure of each compound was visually checked, and 185 compounds were selected so as not to be structurally biased.

で表される化合物が、ＧＡβ依存性アミロイド線維形成に対して阻害活性を有することが示唆された。その後、阻害活性を有した化合物の類似（ｓｉｍｉｌａｒｉｔｙ）検索を行い、ＧＡβ依存性アミロイド線維形成に対する阻害活性を評価した結果、上記一般式（Ｉ）で表される化合物が、ＧＡβ依存性アミロイド線維形成に対して阻害活性を有することが明らかになった。 About the compound narrowed down from the commercial compound by the above, the inhibitory activity with respect to GA (beta) dependence amyloid fibril formation was evaluated in the in vitro reproduction system mentioned later. As a result, the hit compound having the highest priority among those included in the series of hit compound groups is represented by the following general formula (I):

It was suggested that the compound represented by these has inhibitory activity with respect to GAβ-dependent amyloid fibril formation. Thereafter, a similarity search for compounds having inhibitory activity was performed and the inhibitory activity against GAβ-dependent amyloid fibril formation was evaluated. As a result, the compound represented by the above general formula (I) was found to be GAβ-dependent amyloid fibril formation. It became clear that it has inhibitory activity against.

<2. Selection of commercially available compounds (compounds 1 and 13)>
Commercially available compounds having a structure represented by the general formula (I) or commercially available compounds having a structure similar to the structure represented by the general formula (I) are mainly purchased from Enamine, UORSY, Ambinter, etc. did.

<3. Synthesis of novel compounds (compounds 2-12 and 14-29)>
Subsequently, the novel compound which has a structure represented by the said general formula (I) was synthesize | combined. The followings were used as equipment used and measurement conditions.

¹ H NMR spectra were measured using a Bruker Avance or Bruker Varian (400 MHz, 500 MHz, or 600 MHz). TMS (tetramethylsilane) was used as an internal standard.

  LC / MS analysis was performed using a quadrupole mass spectrometer LCMS-2010 (Shimadzu Corporation) (column: Shim-pack XR-ODS (3.0 × 30 mm, 2.2 μm)) using ESI (+) ions. Mode or APCI (+) ion mode and (−) ion mode. Flow rate conditions: 0.3 to 0.8 mL / min, acquisition time: 3 to 5 minutes, detection wavelength: 220 nm, oven temperature: 40 to 50 ° C.

  Prep-HPLC was performed under the following conditions. Silica gel column: Fuji C18 (300 × 25), YMC 250 × 20; wavelength: 220 nm; moving bed A: acetonitrile (0.1% HCl); moving bed B: water; flow rate condition: 25 mL / min, injection amount: 2 mL Run time: 20 minutes; equilibration time: 3 minutes.

<< Synthesis of 4- [benzyl (methyl) amino] -7-chloro-8-methylquinoline-3-carboxylic acid (Compound 1) >>

(1-1) Synthesis of ethyl 7-chloro-4-hydroxy-8-methylquinoline-3-carboxylate (Compound 1-1) Ethyl 7-chloro-4-hydroxy-8-methylquinoline-3-carboxylate ( Compound 1-1) was synthesized with reference to Revista de Chimie (Bucharest, Romania), Volume 61, Issue 8, Pages 745-749, and Journal 2010.

(1-2) Synthesis of ethyl 4,7-dichloro-8-methylquinoline-3-carboxylate (Compound 1-2) Compound 1-1 (13.6 g, 0.52 mmol) was dissolved in thionyl chloride (90 mL). And the solution was reacted at 80 degrees for 3 hours. After the reaction, the reaction solution was concentrated under reduced pressure, and neutralized to pH 8 with sodium hydrogen carbonate aqueous solution to obtain 14 g of ethyl 4,7-dichloro-8-methylquinoline-3-carboxylate (Compound 1-2) as a white solid. (Yield 100%).

(1-3) Synthesis of 4- [benzyl (methyl) amino] -7-chloro-8-methylquinoline-3-carboxylic acid (Compound No. 1) Compound 1-2 (0.85 g, 7,04 mmol) and N , N-diisopropylethylamine (0.91 g, 7.04 mmol) in water (60 mL) was added N-aminobenzylamine (1 g, 3.52 mmol) in acetonitrile (8 mL) and microwaved at 120 degrees for 1 hour. Was irradiated. After the reaction, the reaction solution was evaporated under reduced pressure, a solution obtained by adding ethyl acetate (50 mL) to the residue was washed with saturated brine, and the organic layer was dried over anhydrous sodium sulfate. After filtration, the crude product obtained by concentrating the filtrate was purified by column chromatography to obtain an ester intermediate (1.2 g, 94%).

The obtained ester intermediate (1.2 g, 3.27 mmol) was dissolved in a mixture of methanol (30 mL) and water (10 mL), and sodium hydroxide (0.39 g, 9.81 mmol) was added thereto. After reacting at 50 ° C. for 3 hours, the reaction solution was neutralized to pH 5-6 with dilute hydrochloric acid. The crude product was purified by reverse-layer column chromatography (10-70% methanol / water (0.05% NH ₃ )) to give 4- [benzyl (methyl) amino] -7-chloro-8-methylquinoline-3. -Carboxylic acid (Compound 1) (700 mg, 63%) was obtained.

  Compounds 3-23 and 25-29 were synthesized by the same method as above.

<< Synthesis of 2- (7-chloro-4-{[(4-fluorophenyl) methyl] (methyl) amino} -8-methylquinolin-3-yl) acetic acid (Compound 2) >>

(2-1) Synthesis of dimethyl 2-formylsuccinate (Compound 2-1) To a suspension of metallic sodium (16 g, 690 mmol) in diethyl ether (400 mL), ethyl formate (71.4 g, 965 mmol) and dimethyl succinate (100 g, 575 mmol) was added slowly with stirring. After the reaction was carried out at 40 degrees for 5 hours, water was carefully added to the reaction solution to terminate the reaction. The obtained mixture was washed with diethyl ether (400 mL × 2), the aqueous layer was acidified with 6N hydrochloric acid, and the aqueous layer was extracted with diethyl ether (400 mL × 2). The diethyl ether layer was dried over anhydrous sodium sulfate and then concentrated under reduced pressure of the filtrate to obtain dimethyl 2-formylsuccinate (Compound 2-1) (100 g, 72.3%) as a yellow solid.

(2-2) Synthesis of 2-{[(3-chloro-2-methylphenyl) amino] methylene} dimethyl succinate (Compound 2-2) Compound 2-1 (100 g, 495 mmol) and 3-chloro-2- Methylbenzylamine (49 g, 346 mmol) was dissolved in toluene (300 mL) and reacted at 120 degrees for 5 hours. The reaction solution was concentrated under reduced pressure to obtain dimethyl 2-{[(3-chloro-2-methylphenyl) amino] methylene} succinate (Compound 2-2) (140 g, 87%) as a yellow oily substance.

(2-3) Synthesis of 2- (7-chloro-4-hydroxy-8-methylquinolin-3-yl) acetic acid (Compound 2-3) Compound 2-2 (140 g, 431 mmol) was added to polyphosphoric acid (400 mL). The mixture was further reacted at 140 degrees for 2 hours. After the reaction, the reaction solution was poured into ice water (3000 mL) and stirred for 10 minutes. The precipitate was collected by filtration, washed with water (400 mL), dried and then 2- (7-chloro-4-hydroxy-8-methylquinolin-3-yl) acetic acid (Compound 2-3) (100 g, 92. 4%) was obtained as a pale yellow solid.

(2-4) Synthesis of methyl 2- (7-chloro-4-hydroxy-8-methylquinolin-3-yl) acetate (Compound 2-4) 2- (7-Chloro-4-hydroxy-8-methylquinoline) Concentrated sulfuric acid (70 mL) was added to a methanol (400 mL) solution of -3-yl) acetic acid (compound 2-3) (100 g, 398 mmol), and the mixture was reacted at 80 ° C. overnight. After the reaction, an aqueous sodium hydrogen carbonate solution (2000 mL) was added to the reaction solution and stirred for 1 hour. The resulting precipitate was collected by filtration, washed with water (500 mL), dried, and methyl 2- (7-chloro-4-hydroxy-8-methylquinolin-3-yl) acetate (Compound 2-4) (50 g). 47.5%) as a yellow solid.

(2-5) Synthesis of methyl 2- (4,7-dichloro-8-methylquinolin-3-yl) acetate (Compound 2-5) Compound 2-4 (50 g, 188 mmol) was converted to phosphorus oxychloride (250 mL). Was added under ice cooling. After reacting at 80 ° C. for 16 hours, the reaction solution was ice-cooled, and then poured into ice water (2500 g) with vigorous stirring. The mixture was neutralized with 4M aqueous sodium hydroxide solution and extracted with ethyl acetate (400 mL × 3). The organic layer was washed with saturated brine (600 mL), dried and filtered over anhydrous sodium sulfate, and the filtrate was concentrated to obtain a crude product. This was purified by column chromatography to obtain methyl 2- (4,7-dichloro-8-methylquinolin-3-yl) acetate (Compound 2-5) (15 g, 28%) as a yellow solid.

(2-6) Synthesis of 2- (7-chloro-4-{[(4-fluorophenyl) methyl] (methyl) amino} -8-methylquinolin-3-yl) acetic acid (Compound No. 2) Compound 2- To a solution of 5 (15 g, 14.8 mmol) in methanol (140 mL) was added THF (60 mL), water (50 mL) and lithium hydroxide (11.1 g, 265 mmol), and the mixture was stirred at room temperature for 1 hour. After the reaction, the mixture was concentrated under reduced pressure, and the resulting residue was diluted with water (100 mL), acidified with 3N hydrochloric acid to pH 3-4, and stirred for 10 minutes. The deposited precipitate was collected by filtration, washed with water and then dried to obtain a carboxylic acid intermediate (10.9 g, 76.4%) as yellow crystals.

  Subsequently, a suspension of carboxylic acid intermediate (10.9 g, 40.5 mmol) in n-butanol (90 mL) was stirred with N-methyl-4-fluorobenzylamine (22.5 g, 162 mmol) and N, N-diisopropylethylamine (21 g, 162 mmol) was added. The reaction was carried out at 140 degrees for 20 hours. After the reaction, the mixture was concentrated under reduced pressure, and ethyl acetate (60 mL) was added to the resulting residue for dilution. This solution was washed with water (30 mL × 2), dried and filtered with anhydrous sodium sulfate, and concentrated, and the crude product obtained by concentration was purified by silica gel chromatography to give 2- (7-chloro-4-{[( 4-Fluorophenyl) methyl] (methyl) amino} -8-methylquinolin-3-yl) acetic acid (Compound 2) (2.53 g, 16.8%) was obtained.

  Compound 24 was synthesized by the same method as above.

Table 2 shows compounds 1-29.

<3. Evaluation of amyloid fibril formation inhibitory activity by thioflavin T (ThT) assay>
The inhibitory activity of the compounds 1 to 29 on GAβ-dependent amyloid fibril formation was evaluated by ThT assay.

(1) Preparation of Aβ solution Human Aβ1-40 (cat. # 4307-v, manufactured by Peptide Institute) was dissolved in 0.02% ammonia solution (400 μM), and centrifuged at 100,000 rpm for 3 hours (S110AT Rotor manufactured by Hitachi). About two thirds of the upper layer of the supernatant was collected and the protein concentration was measured. The Aβ solution was aliquoted and stored at −80 ° C. until use. Thawed immediately before use and diluted to 75 μM with phosphate buffer (PBS: cat. # 10010, Life Technologies).

(2) Preparation of GM1-containing liposome Using a chloroform / methanol mixture (1: 1) as a solvent, a mixed solution of 100 mM cholesterol and sphingomyelin (both manufactured by Sigma-Aldrich) was prepared. In this mixed solution, GM1 (cat. # 1061, manufactured by Matreya) was dissolved to a concentration of 50 mM. This lipid mixture solution was aliquoted into glass test tubes and stored at −20 ° C. until use. After melting before use, chloroform / methanol mixture (1: 1) was added and mixed, and then the solvent was removed by applying nitrogen gas to the suspension. The dried lipid mixture was resuspended in PBS so that the GM1 concentration was 2.5 mM, and freeze-thawed 5 times using liquid nitrogen. This lipid suspension was centrifuged at 15,000 rpm for 10 minutes, and the precipitate was resuspended in PBS so that GM1 was 2.5 mM. Using an ultrasonic crusher (XL-2000, manufactured by MISONIX) equipped with a microchip, sonication for 10 seconds was repeated on ice until this suspension became transparent to prepare GM1-containing liposomes. PBS was added immediately before use to dilute the GM1 concentration to 750 μM.

(3) Preparation of Compound Solution Compounds 1 to 29 were prepared as 10 mM and 2 mM solutions in DMSO, respectively. Immediately before use, it was diluted to 75 μM or 15 μM with PBS.

(4) GAβ-dependent amyloid fibril formation In a 96-well microplate, 75 μM of the Aβ solution prepared in (1) above and 5 μl of the compound solution prepared in (3) or 0.75% DMSO were mixed, Incubated for 10 minutes at room temperature. Next, 5 μl of 750 μM GM1-containing liposome prepared in (2) was added to each well and incubated at 37 ° C. for 18 hours.

(5) ThT assay ThT (manufactured by Sigma-Aldrich) was prepared to 5 μM using 50 mM glycine-NaOH buffer (pH 8.5). Immediately before the measurement, 10 μl from each well in (4) was transferred to a 96-well black plate. Using a multiplate reader (ARVO-HTS, manufactured by PerkinElmer), 50 μl of 5 μM ThT solution was added to each well from an injector, and then optimum fluorescence of ThT bound to amyloid fibrils (excitation wavelength: 430 nm, fluorescence wavelength) 490 nm).

  The fluorescence intensity of ThT indicates the degree of amyloid fibril formation. By determining the ratio of the ThT fluorescence intensity when the compound was added to the ThT fluorescence intensity when no compound was added, the inhibition rate of amyloid fibril formation by the compound was calculated. When the final concentration of the compound was 25 μM, the inhibition rate was evaluated as B, and the inhibition rate as 40% or more and less than 80% was evaluated as B.

  In Table 3, evaluation about the amyloid fibril formation inhibitory activity of the compounds 1-29 is shown. Compounds 1 to 29 were all confirmed to have inhibitory activity against amyloid fibril formation.

Claims (8)

Formula (I):

[Where:
X represents — (CH ₂ ) _n —.
(In the formula, n is an integer of 0 to 3)
And
Y is a single bond or a divalent one having 1 to 4 atoms, the main chain being composed of one or more atoms selected from the group consisting of carbon atoms, nitrogen atoms and oxygen atoms. A linking group,
Ar is a substituted or unsubstituted phenyl group or pyridyl group;
R ₁ is a substituted or unsubstituted C _1-6 alkyl group or a substituted or unsubstituted C _3-7 cycloalkyl group,
Here, Ar and R ₁ , Ar and Y, or R ₁ and Y may jointly form a ring,
R ₂ , R ₃ and R ₄ are each independently a hydrogen atom, a C _1-20 alkyl group, a C _1-20 hydroxyalkyl group, a C _1-20 alkoxy group or a halogen atom]
Or a pharmaceutically acceptable salt thereof or a solvate thereof, an amyloid fibril formation inhibitor. Said compound has the general formula (Ia):

[Where:
Y is a single bond or a divalent group selected from the group consisting of a substituted or unsubstituted C _1-20 alkyl group, an amide group, an amine group, an ether group, and a substituted or unsubstituted C _3-20 cycloalkyl group. A linking group of
Ar is a substituted or unsubstituted phenyl group,
R ₁ is a substituted or unsubstituted C _1-4 alkyl group or a substituted or unsubstituted C _3-6 cycloalkyl group;
Here, Ar and R ₁ , Ar and Y, or R ₁ and Y may jointly form a ring,
R ₂ and R ₃ are each independently a hydrogen atom, a C _1-20 alkyl group or a halogen atom]
The amyloid fibril formation inhibitor of Claim 1 represented by these.   Inhibition of amyloid fibril formation, comprising one or more compounds selected from the group consisting of compounds Nos. 1-29 described in Table 1A and Table 1B, or pharmaceutically acceptable salts thereof, or solvates thereof Agent.   4. The amyloid according to claim 3, wherein the compound is selected from the group consisting of compounds Nos. 1, 2, 4, 7, 9, 10, 13, 15, 18, 21, 23-25, 27 and 28. Fibrosis inhibitor.   The amyloid fibril formation inhibitor according to claim 3, wherein the compound is selected from the group consisting of compounds of Compound Nos. 1, 2, 7, 9, 23 and 25.   The therapeutic agent or preventive agent of a neurodegenerative disease containing the amyloid fibril formation inhibitor of any one of Claims 1-5.   The therapeutic or prophylactic agent according to claim 6, wherein the neurodegenerative disease is Alzheimer's disease.   A compound selected from the group consisting of the compounds of Compound Nos. 1 to 29 described in Table 1A and Table 1B, or a pharmaceutically acceptable salt thereof, or a solvate thereof.