JP2009203166A

JP2009203166A - Diagnostic composition containing diphenyloxadiazole derivative

Info

Publication number: JP2009203166A
Application number: JP2008043750A
Authority: JP
Inventors: Masahiro Ono; 正博小野; Morio Nakayama; 守雄中山
Original assignee: Nagasaki University NUC
Current assignee: Nagasaki University NUC
Priority date: 2008-02-26
Filing date: 2008-02-26
Publication date: 2009-09-10

Abstract

<P>PROBLEM TO BE SOLVED: To provide a compound that exhibits high bonding specificity to an amyloid β protein and high permeability through blood brain barriers as well as rapid disappearance from a region other than intracerebral senile plaque. <P>SOLUTION: The diagnostic composition for amyloid protein-related diseases comprises a compound represented by general formula (I) (wherein R<SP>1</SP>is a 4-fluorophenyl group or the like; and R<SP>2</SP>is a 4-methoxyphenyl group or the like), or a compound obtained by labeling the compound with a radioactive nuclide, or a pharmaceutically acceptable salt of these compounds. <P>COPYRIGHT: (C)2009,JPO&INPIT

Description

本発明は、ジフェニルオキサジアゾール（DPOD）誘導体を含有する診断用組成物に関する。本発明の組成物は、アルツハイマー病などのアミロイド性タンパク質関連疾患の診断に用いることができる。 The present invention relates to a diagnostic composition containing a diphenyloxadiazole (DPOD) derivative. The composition of the present invention can be used for diagnosis of amyloid protein-related diseases such as Alzheimer's disease.

近年の急速な高齢化に伴い、アルツハイマー病(AD)をはじめとする痴呆性疾患の増加が大きな社会問題のひとつになっている。現在、ADの臨床診断法には、長谷川式、ADAS、MMSEがあり、いずれもADが疑われる個体の認知機能の低下を定量的に評価する方法が一般的に用いられる。この他画像診断法（MRI, CT等）が補助的に用いられるが、これらの診断法ではADを確定診断するには不十分であり、確定診断には生前における脳の生検、死後脳の病理組織学的検査において、老人斑と神経原繊維の出現を確認することが必要である。したがって、現在の診断方法では、広範な脳障害が生じる前の早期段階でADを診断するのは困難である。これまでにADの生物学的診断マーカーとしていくつかの報告があるが、臨床上実用的なものはいまだ開発されていない。このような状況下、ADの早期診断に対する社会的要求は高く、その早急な開発が強く望まれている。 With the recent rapid aging of society, an increase in dementia diseases such as Alzheimer's disease (AD) has become one of the major social problems. Currently, there are Hasegawa's method, ADAS, and MMSE as clinical diagnostic methods for AD, and all of them generally use a method for quantitatively evaluating the decline in cognitive function of an individual suspected of having AD. Other diagnostic imaging methods (MRI, CT, etc.) are used supplementarily, but these diagnostic methods are not sufficient for definitive diagnosis of AD. For definitive diagnosis, biopsy of the brain before birth, postmortem brain In histopathological examination, it is necessary to confirm the appearance of senile plaques and neurofibrils. Therefore, with current diagnostic methods, it is difficult to diagnose AD at an early stage before extensive brain damage occurs. To date, there have been several reports as biological diagnostic markers for AD, but clinically practical ones have not yet been developed. Under such circumstances, there is a high social demand for early diagnosis of AD, and its rapid development is strongly desired.

老人斑はADの最も特徴的な脳病変であり、その主構成成分はβシート構造をとったアミロイドβタンパク質である。体外からの老人斑の画像化はADの有効な診断法の確立につながると考えられるが、画像化には、アミロイドβタンパク質と特異的に結合するプローブ化合物が必要である。これまでに、プローブ化合物としてコンゴーレッドおよびチオフラビンTを母体構造とする誘導体が、いくつか報告されているが（特開2004-250407号公報、特開2004-250411号公報、W.E.Klunk et al., Annals of Neurology Vol55 No.3 March 2004 306-319）、アミロイドβタンパク質に対する結合特異性が低いこと、血液脳関門の透過性が低いこと、脳内での非特異的結合によりクリアランスが遅いことなど問題が少なくない。それゆえ、報告されたこれらの化合物は未だアミロイドが蓄積する疾患の診断において実用化されていないのが現状である。 Senile plaque is the most characteristic brain lesion of AD, and its main component is amyloid β protein with β sheet structure. Imaging of senile plaques from outside the body is thought to lead to the establishment of an effective diagnostic method for AD, but imaging requires a probe compound that specifically binds to amyloid β protein. Until now, several derivatives having a parent structure of Congo red and thioflavin T as probe compounds have been reported (JP 2004-250407 A, JP 2004-250411 A, WEKlunk et al., Annals of Neurology Vol55 No.3 March 2004 306-319), low binding specificity for amyloid β protein, low permeability of blood-brain barrier, slow clearance due to nonspecific binding in the brain There are many. Therefore, at present, these reported compounds have not been put into practical use in the diagnosis of diseases in which amyloid accumulates.

以上のような既存のプローブ化合物の問題点を解決するため、本発明者らはフラボノイド系化合物であるフラボン、カルコン、オーロンを基本骨格としたプローブ化合物を開発し、出願を行った（特許文献１、特許文献２）。 In order to solve the problems of the existing probe compounds as described above, the present inventors have developed and applied for a probe compound having flavone, chalcone, or aurone as basic skeletons (Patent Document 1). Patent Document 2).

国際公開第2006/057323号パンフレットInternational Publication No. 2006/057323 Pamphlet 国際公開第2007/136059号パンフレットInternational Publication No. 2007/136059 Pamphlet

本発明は、以上のような技術的背景のもとになされたものであり、アミロイドβタンパク質に対する高い結合特異性、高い血液脳関門の透過性、脳内老人斑以外の部位からの速やかな消失性を併せ持つ化合物を提供することを目的とする。 The present invention has been made based on the technical background as described above, and has high binding specificity for amyloid β protein, high permeability of the blood brain barrier, and rapid disappearance from sites other than brain senile plaques. An object is to provide a compound having both properties.

本発明者は、上記課題を解決するため鋭意検討を重ねた結果、フラボノイド系化合物とは異なる、DPODを基本骨格とする化合物が、アミロイドβタンパク質への高い結合性、脳移行性と速やかなクリアランスを示し、さらに、アルツハイマー病患者脳組織切片を用いた蛍光染色において、老人斑への選択的結合性を示すことを見出し、本発明を完成するに至った。 As a result of intensive studies to solve the above problems, the present inventor found that a compound having DPOD as a basic skeleton, which is different from a flavonoid compound, has a high binding ability to amyloid β protein, a brain transportability and a rapid clearance. Furthermore, it was found that the fluorescent staining using brain tissue sections of Alzheimer's disease patients showed selective binding to senile plaques, and the present invention was completed.

即ち、本発明は、以下の（１）〜（１３）を提供するものである。 That is, the present invention provides the following (1) to (13).

（１）一般式（Ｉ） (1) General formula (I)

〔式中、Ｒ^１は下記置換基群Ａから選ばれた１若しくは２以上の置換基によって置換されていてもよいフェニル基を表し、Ｒ^２は下記置換基群Ｂから選ばれた１若しくは２以上の置換基によって置換されていてもよいフェニル基を表し、置換基群Ａは、ハロゲン原子、式：−（ＣＨ_２ＣＨ_２Ｏ）_ｎ−Ｆ [式中、ｎは１〜１０の整数を表す。] で表される基、及び式：−（ＣＨ_２ＣＨ_２Ｏ）_ｎ−ＯＨ [式中、ｎは１〜１０の整数を表す。] で表される基からなる群であり、置換基群Ｂは、ハロゲン原子、ジメチルアミノ基、メチルアミノ基、アミノ基、メトキシ基、水酸基、式：−（ＣＨ_２ＣＨ_２Ｏ）_ｎ−Ｆ [式中、ｎは１〜１０の整数を表す。] で表される基、及び式：−（ＣＨ_２ＣＨ_２Ｏ）_ｎ−ＯＨ [式中、ｎは１〜１０の整数を表す。] で表される基からなる群である。〕
又は一般式（II） [Wherein, R ¹ represents a phenyl group which may be substituted with one or more substituents selected from the following substituent group A, and R ² represents 1 or 2 selected from the following substituent group B represents a more substituents which may be a phenyl group substituted by, substituent group a is a halogen atom, in the formula _{_{_{:-( CH 2 CH 2 O) n}}} -F [ wherein, n represents an integer of 1 to 10 To express. And a group represented by the formula: — (CH ₂ CH ₂ O) _n —OH [wherein n represents an integer of 1 to 10. The substituent group B is a halogen atom, a dimethylamino group, a methylamino group, an amino group, a methoxy group, a hydroxyl group, a formula: — (CH ₂ CH ₂ O) _n —F. [Wherein n represents an integer of 1 to 10. And a group represented by the formula: — (CH ₂ CH ₂ O) _n —OH [wherein n represents an integer of 1 to 10. ] Is a group consisting of groups represented by ]
Or general formula (II)

〔式中、Ｒ^１及びＲ^２は前記と同意義を示す。〕
で表される化合物若しくは前記化合物を放射性核種で標識した化合物、又はこれらの化合物の医薬上許容される塩を含有するアミロイド性タンパク質関連疾患診断用組成物。 [Wherein, R ¹ and R ² are as defined above. ]
Or a compound obtained by labeling said compound with a radionuclide, or a pharmaceutically acceptable salt of these compounds.

（２）アミロイド性タンパク質関連疾患が、アミロイドβタンパク質関連疾患である（１）に記載のアミロイド性タンパク質関連疾患診断用組成物。 (2) The amyloid protein-related disease diagnosis composition according to (1), wherein the amyloid protein-related disease is an amyloid β protein-related disease.

（３）アミロイド性タンパク質関連疾患が、アルツハイマー病である（１）に記載のアミロイド性タンパク質関連疾患診断用組成物。 (3) The amyloid protein-related disease diagnostic composition according to (1), wherein the amyloid protein-related disease is Alzheimer's disease.

（４）一般式（Ｉ）及び（II）におけるＲ^１が、フッ素原子又はヨウ素原子で置換されたフェニル基である（１）乃至（３）のいずれかに記載のアミロイド性タンパク質関連疾患診断用組成物。 (4) For diagnosing amyloid protein-related diseases according to any one of (1) to (3), wherein R ¹ in the general formulas (I) and (II) is a phenyl group substituted with a fluorine atom or an iodine atom Composition.

（５）一般式（Ｉ）及び（II）におけるＲ^１が、４−フルオロフェニル基、又は４−ヨードフェニル基である（１）乃至（３）のいずれかに記載のアミロイド性タンパク質関連疾患診断用組成物。 (5) Diagnosis of amyloid protein-related disease according to any one of (1) to (3), wherein R ¹ in the general formulas (I) and (II) is a 4-fluorophenyl group or a 4-iodophenyl group Composition.

（６）一般式（Ｉ）及び（II）におけるＲ^２が、ジメチルアミノ基、メチルアミノ基、アミノ基、メトキシ基、又は水酸基で置換されたフェニル基である（１）乃至（５）のいずれかに記載のアミロイド性タンパク質関連疾患診断用組成物。 (6) Any of (1) to (5), wherein R ² in general formulas (I) and (II) is a dimethylamino group, a methylamino group, an amino group, a methoxy group, or a phenyl group substituted with a hydroxyl group A composition for diagnosing amyloid protein-related diseases according to claim 1.

（７）一般式（Ｉ）及び（II）におけるＲ^２が、ジメチルアミノ基、メチルアミノ基、アミノ基、又はメトキシ基で置換されたフェニル基である（１）乃至（５）のいずれかに記載のアミロイド性タンパク質関連疾患診断用組成物。 (7) In any one of (1) to (5), R ² in the general formulas (I) and (II) is a phenyl group substituted with a dimethylamino group, a methylamino group, an amino group, or a methoxy group. The composition for amyloid protein-related disease diagnosis as described.

（８）一般式（Ｉ）及び（II）におけるＲ^２が、メトキシ基で置換されたフェニル基である（１）乃至（５）のいずれかに記載のアミロイド性タンパク質関連疾患診断用組成物。 (8) The amyloid protein-related disease diagnostic composition according to any one of (1) to (5), wherein R ² in the general formulas (I) and (II) is a phenyl group substituted with a methoxy group.

（９）一般式（Ｉ）及び（II）におけるＲ^２が、４−メトキシフェニル基である（１）乃至（５）のいずれかに記載のアミロイド性タンパク質関連疾患診断用組成物。 (9) The amyloid protein-related disease diagnostic composition according to any one of (1) to (5), wherein R ² in the general formulas (I) and (II) is a 4-methoxyphenyl group.

（１０）放射性核種が、陽電子放出核種である（１）乃至（９）のいずれかに記載のアミロイド性タンパク質関連疾患診断用組成物。 (10) The composition for diagnosing amyloid protein-related disease according to any one of (1) to (9), wherein the radionuclide is a positron emitting nuclide.

（１１）放射性核種が、γ線放出核種である（１）乃至（９）のいずれかに記載のアミロイド性タンパク質関連疾患診断用組成物。 (11) The composition for diagnosing amyloid protein-related disease according to any one of (1) to (9), wherein the radionuclide is a γ-ray-emitting nuclide.

（１２）アミロイド性タンパク質関連疾患のモデル動物に被験物質を投与する工程、前記モデル動物に（１）乃至（１１）のいずれかに記載のアミロイド性タンパク質関連疾患診断用組成物を投与する工程、及び前記モデル動物の脳中に含まれる一般式（Ｉ）又は（II）で表される化合物の分布又は量を調べる工程を含むアミロイド性タンパク質関連疾患の治療薬又は予防薬のスクリーニング方法。 (12) a step of administering a test substance to a model animal of amyloid protein-related disease, a step of administering to the model animal the composition for diagnosing amyloid protein-related disease according to any one of (1) to (11), And a screening method for a therapeutic or prophylactic agent for amyloid protein-related diseases, comprising the step of examining the distribution or amount of the compound represented by the general formula (I) or (II) contained in the brain of the model animal.

（１３）アミロイド性タンパク質関連疾患のモデル動物に前記疾患の治療薬又は予防薬を投与する工程、前記モデル動物に（１）乃至（１１）のいずれかに記載のアミロイド性タンパク質関連疾患診断用組成物を投与する工程、及び前記モデル動物の脳中に含まれる一般式（Ｉ）又は（II）で表される化合物の分布又は量を調べる工程を含むアミロイド性タンパク質関連疾患の治療薬又は予防薬の評価方法。 (13) A step of administering a therapeutic or prophylactic agent for the disease to a model animal of amyloid protein-related disease, the composition for diagnosing amyloid protein-related disease according to any one of (1) to (11), to the model animal A therapeutic or prophylactic agent for amyloid protein-related diseases comprising a step of administering a product, and a step of examining the distribution or amount of the compound represented by the general formula (I) or (II) contained in the brain of the model animal Evaluation method.

一般式（Ｉ）又は（II）で表される化合物は、アミロイドβタンパク質に対し高い結合特異性を持ち、また、脳内老人斑以外の部位からの速やかに消失する性質を持つので、アルツハイマー病の診断に有用である。 The compound represented by the general formula (I) or (II) has a high binding specificity for amyloid β protein, and also has a property of rapidly disappearing from a site other than cerebral senile plaques. Therefore, Alzheimer's disease Useful for diagnosis.

以下、本発明を詳細に説明する。 Hereinafter, the present invention will be described in detail.

本発明において、「ハロゲン原子」とは、例えば、フッ素原子、塩素原子、臭素原子、ヨウ素原子である。 In the present invention, the “halogen atom” is, for example, a fluorine atom, a chlorine atom, a bromine atom, or an iodine atom.

式：−（ＣＨ_２ＣＨ_２Ｏ）_ｎ−Ｆにおいてｎは、好適には１〜３である。 N In the formula _{_{_{:-( CH 2 CH 2 O) n}}} -F is preferably 1 to 3.

式：−（ＣＨ_２ＣＨ_２Ｏ）_ｎ−ＯＨにおいてｎは、好適には１〜３である。 In the formula: — (CH ₂ CH ₂ O) _n —OH, n is preferably 1 to 3.

一般式（Ｉ）及び（II）で表される化合物のいずれも本発明の診断用組成物として利用できるが、脳への移行性が高いことから、一般式（II）で表される化合物の方が好ましい。 Any of the compounds represented by the general formulas (I) and (II) can be used as the diagnostic composition of the present invention, but because of its high ability to migrate to the brain, the compounds represented by the general formula (II) Is preferred.

Ｒ^１のフェニル基の置換基としては、フッ素原子、ヨウ素原子が好ましい。フェニル基の置換基の位置は、２位、３位であってもよいが、４位であることが好ましい。Ｒ^１の具体例としては、４−フルオロフェニル基、４−ヨードフェニル基などを挙げることができる。 The substituent for the phenyl group of R ¹ is preferably a fluorine atom or an iodine atom. The position of the substituent of the phenyl group may be the 2-position or the 3-position, but is preferably the 4-position. Specific examples of R ¹ include 4-fluorophenyl group and 4-iodophenyl group.

Ｒ^２のフェニル基の置換基としては、ジメチルアミノ基、メチルアミノ基、アミノ基、メトキシ基、又は水酸基が好ましい。これらの中でも、アミロイドβタンパク質に対する結合性が高くなることから、ジメチルアミノ基、メチルアミノ基、アミノ基、又はメトキシ基が好ましく、メトキシ基が最も好ましい。フェニル基の置換基の位置は、２位、３位であってもよいが、４位であることが好ましい。Ｒ^２の具体例としては、４−ジメチルアミノフェニル基、４−メチルアミノフェニル基、４−アミノフェニル基、４−メトキシフェニル基、４−ヒドロキシフェニル基、２―メトキシフェニル基、３−メトキシフェニル基などを挙げることができ、これらの中でも、２―メトキシフェニル基、３−メトキシフェニル基、４−メトキシフェニル基が好ましく、４−メトキシフェニル基が最も好ましい。 Examples of the substituent of the phenyl group of R ^2, dimethylamino group, methylamino group, an amino group, a methoxy group, or a hydroxyl group are preferred. Among these, a dimethylamino group, a methylamino group, an amino group, or a methoxy group is preferable because the binding property to amyloid β protein is high, and a methoxy group is most preferable. The position of the substituent of the phenyl group may be the 2-position or the 3-position, but is preferably the 4-position. Specific examples of R ² include 4-dimethylaminophenyl group, 4-methylaminophenyl group, 4-aminophenyl group, 4-methoxyphenyl group, 4-hydroxyphenyl group, 2-methoxyphenyl group, 3-methoxyphenyl. Among them, 2-methoxyphenyl group, 3-methoxyphenyl group and 4-methoxyphenyl group are preferable, and 4-methoxyphenyl group is most preferable.

一般式(Ｉ)で表される化合物のうち代表的なものを表１〜８に示し、一般式（II）で表される化合物のうち代表的なものを表９〜１６に示す。なお、表中の「FX-」は、式：−（ＣＨ_２ＣＨ_２Ｏ）−Ｆで表される基を示し、「FX2-」は、式：−（ＣＨ_２ＣＨ_２Ｏ）_２−Ｆで表される基を示し、「FX3-」は、式：−（ＣＨ_２ＣＨ_２Ｏ）_３−Ｆで表される基を示し、「HOX-」は、式：−（ＣＨ_２ＣＨ_２Ｏ）−ＯＨで表される基を示し、「HOX2-」は、式：−（ＣＨ_２ＣＨ_２Ｏ）_２−ＯＨで表される基を示し、「HOX3-」は、式：−（ＣＨ_２ＣＨ_２Ｏ）_３−ＯＨで表される基を示す。 Representative compounds of the general formula (I) are shown in Tables 1 to 8, and typical compounds of the general formula (II) are shown in Tables 9 to 16. In the table, “FX-” represents a group represented by the formula: — (CH ₂ CH ₂ O) —F, and “FX2-” represents the formula: — (CH ₂ CH ₂ O) ₂ —F. “FX3-” represents a group represented by the formula: — (CH ₂ CH ₂ O) ₃ —F, and “HOX—” represents a formula: — (CH ₂ CH ₂ O ) —OH represents a group, “HOX2-” represents a group represented by the formula: — (CH ₂ CH ₂ O) ₂ —OH, and “HOX3-” represents a formula: — (CH ₂ CH ₂ _O) a group represented by 3 -OH.

上記化合物のうちで、好ましい化合物として、I-4、I-9、I-14、I-19、I-24、I-29、I-34、I-39、I-44、I-49、I-54、I-59、I-64、I-69、I-74、I-79、I-84、I-89、II-4、II-9、II-14、II-19、II-24、II-29、II-34、II-39、II-44、II-49、II-54、II-59、II-64、II-69、II-74、II-79、II-84、II-89を挙げることができ、より好ましい化合物として、I-14、I-29、I-44、I-59、I-74、I-89、II-14、II-29、II-44、II-59、II-74、II-89を挙げることができ、更により好ましい化合物として、I-44、I-89、II-44、II-89を挙げることができる。 Among the above compounds, preferred compounds include I-4, I-9, I-14, I-19, I-24, I-29, I-34, I-39, I-44, I-49, I-54, I-59, I-64, I-69, I-74, I-79, I-84, I-89, II-4, II-9, II-14, II-19, II- 24, II-29, II-34, II-39, II-44, II-49, II-54, II-59, II-64, II-69, II-74, II-79, II-84, II-89 can be mentioned, and more preferred compounds include I-14, I-29, I-44, I-59, I-74, I-89, II-14, II-29, II-44, II-59, II-74 and II-89 can be mentioned, and still more preferred compounds include I-44, I-89, II-44 and II-89.

一般式（Ｉ）又は（II）で表される化合物は、後述する実施例の記載、及びSantagada V. et al., Bioorganic & Medicinal Chemistry Letters, 2004, 14, 4491-3, Mahboobeh B et al., Tetrahedron Letter, 2006, 47, 6983-6などの記載に従って合成することができる。 The compounds represented by the general formula (I) or (II) are described in the Examples described later, and Santagada V. et al., Bioorganic & Medicinal Chemistry Letters, 2004, 14, 4491-3, Mahboobeh B et al. , Tetrahedron Letter, 2006, 47, 6983-6.

一般式（Ｉ）又は（II）で表される化合物は、標識物質によって標識されていることが好ましい。標識物質としては、蛍光物質、アフィニティー物質などを使用してもよいが、放射性核種を使用するのが好しい。標識に用いる放射性核種の種類は特に限定されず、使用の態様によって適宜決めることができる。例えば、一般式（Ｉ）又は（II）で表される化合物をコンピューター断層撮影法（SPECT）による診断に使用する場合、放射性核種は^９９ｍＴｃ、^１１１Ｉｎ、^６７Ｇａ、^２０１Ｔｌ、^１２３Ｉ、^１３３Ｘｅ（好適には、^９９ｍＴｃ、^１２３Ｉ）などのγ線放出核種を使用することができる。また、陽電子断層撮影法（PET）による診断に使用する場合には、^１１Ｃ、^１３Ｎ、^１５Ｏ、^１８Ｆ、^６２Ｃｕ、^６８Ｇａ、^７６Ｂｒ（好適には、^１１Ｃ、^１３Ｎ、^１５Ｏ、^１８Ｆ）などの陽電子放出核種を使用することができる。また、一般式（Ｉ）又は（II）で表される化合物をヒト以外の動物に投与する場合には、より半減期の長い放射性核種、例えば、^１２５Ｉなどを使用してもよい。放射性核種は、一般式（Ｉ）又は（II）で表される化合物の分子中に含まれる形でもよく、また、一般式（Ｉ）又は（II）で表される化合物に結合する形であってもよい。 The compound represented by the general formula (I) or (II) is preferably labeled with a labeling substance. As the labeling substance, a fluorescent substance or an affinity substance may be used, but it is preferable to use a radionuclide. The kind of radionuclide used for labeling is not particularly limited, and can be appropriately determined depending on the mode of use. For example, when the compound represented by the general formula (I) or (II) is used for diagnosis by computed tomography (SPECT), the radionuclide is ^99m Tc, ¹¹¹ In, ⁶⁷ Ga, ²⁰¹ Tl, ¹²³ I, ^133. Γ-ray emitting nuclides such as Xe (preferably ^99m Tc, ¹²³ I) can be used. When used for diagnosis by positron emission tomography (PET), ¹¹ C, ¹³ N, ¹⁵ O, ¹⁸ F, ⁶² Cu, ⁶⁸ Ga, ⁷⁶ Br (preferably ¹¹ C, ¹³ N, ¹⁵ Positron emitting nuclides such as O, ¹⁸ F) can be used. In addition, when the compound represented by the general formula (I) or (II) is administered to an animal other than a human, a radionuclide having a longer half-life, for example, ¹²⁵ I may be used. The radionuclide may be included in the molecule of the compound represented by the general formula (I) or (II), or may be bound to the compound represented by the general formula (I) or (II). May be.

一般式（Ｉ）又は（II）で表される化合物に放射性核種を結合させる方法は、各放射性核種において一般的に用いられている方法でよい。また、一般式（Ｉ）又は（II）で表される化合物に放射性核種を結合させる場合、放射性核種のみを結合させてもよいが、他の物質と結合した状態の放射性核種を結合させてもよい。前述した^９９ｍＴｃは、通常、錯体の形で被標識化合物に結合させるので、一般式（Ｉ）又は（II）で表される化合物に結合させる場合も、^９９ｍＴｃを含む錯体を結合させてもよい。^９９ｍＴｃを含む錯体としては、２−ヒドラジノピリジンを含む錯体（Liu S et al, Bioconjug Chem. 1996 Jan-Feb;7(1):63-71.）、Ｎ−（２−メルカプトエチル）−２−〔（２−メルカプトエチル）アミノ〕−アセトアミドを含む錯体（Zhen W et al, J Med Chem. 1999 Jul 29;42(15):2805-15.）、２，２’−（１，２−エタンジイルジイミノ）ビスエタンチオールを含む錯体（Oya S et al, Nucl Med Biol. 1998 Feb;25(2):135-40.）、トリカルボニル錯体（Schibli R et al, Bioconjug Chem. 2000 May-Jun;11(3):345-51）などを例示できる。 The method of binding the radionuclide to the compound represented by the general formula (I) or (II) may be a method generally used in each radionuclide. In addition, when a radionuclide is bound to the compound represented by the general formula (I) or (II), only the radionuclide may be bound, or a radionuclide that is bound to another substance may be bound. Good. ^99m Tc described above is usually bonded to a labeled compound in the form of a complex. Therefore, even if it is bonded to a compound represented by the general formula (I) or (II), a complex containing ^99m Tc may be bonded. Good. ^As a complex containing ^99m Tc, a complex containing 2-hydrazinopyridine (Liu S et al, Bioconjug Chem. 1996 Jan-Feb; 7 (1): 63-71.), N- (2-mercaptoethyl)- Complexes containing 2-[(2-mercaptoethyl) amino] -acetamide (Zhen W et al, J Med Chem. 1999 Jul 29; 42 (15): 2805-15.), 2,2 ′-(1,2 -Ethanediyldiimino) bisethanethiol-containing complexes (Oya S et al, Nucl Med Biol. 1998 Feb; 25 (2): 135-40.), Tricarbonyl complexes (Schibli R et al, Bioconjug Chem. 2000 May -Jun; 11 (3): 345-51).

一般式（Ｉ）又は（II）で表される化合物の代わりに、医薬上許容される塩を使用することも可能である。医薬上許容される塩としては、アルカリ金属塩（ナトリウム塩、カリウム塩、リチウム塩）、アルカリ土類金属塩（カルシウム塩、マグネシウム塩）、硫酸塩、塩酸塩、硝酸塩、リン酸塩などを例示できる。 It is also possible to use a pharmaceutically acceptable salt in place of the compound represented by the general formula (I) or (II). Examples of pharmaceutically acceptable salts include alkali metal salts (sodium salt, potassium salt, lithium salt), alkaline earth metal salts (calcium salt, magnesium salt), sulfate, hydrochloride, nitrate, phosphate, etc. it can.

本発明の組成物はアミロイド性タンパク質関連疾患の診断に用いられる。ここで、「アミロイド性タンパク質関連疾患」とは、アミロイドβタンパク質関連疾患やプリオン病などを意味する。「アミロイドβタンパク質関連疾患」とは、アミロイドβタンパク質の蓄積によって起きる疾患をいい、主にアルツハイマー病を意味するが、ダウン症候群、オランダ型アミロイドーシスを伴う遺伝性脳出血症（hereditary cerebral hemorrhage with amyloidosis─Dutch type: HCHWA-D）などの疾患も含まれる。また、「プリオン病」とは、異常型プリオンタンパク質によって起きる疾患をいい、クロイツフェルド-ヤコブ病(CJD)やウシ海綿状脳症(BSE)を含む。本発明のアミロイド性タンパク質関連疾患には、一般には「疾患」と認識されない疾患の前駆症状も含まれる。このような疾患の前駆症状としては、アルツハイマー病の発症前にみられる軽度認知障害（MCI）などを例示できる。 The composition of the present invention is used for diagnosis of amyloid protein-related diseases. Here, “amyloid protein-related disease” means amyloid β protein-related disease, prion disease and the like. “Amyloid β protein-related disease” refers to a disease caused by accumulation of amyloid β protein, which mainly means Alzheimer's disease, but hereditary cerebral hemorrhage with amyloidosis (Dutch) Type: HCHWA-D) and other diseases are also included. “Prion disease” refers to a disease caused by an abnormal prion protein, and includes Creutzfeld-Jakob disease (CJD) and bovine spongiform encephalopathy (BSE). The amyloid protein-related diseases of the present invention include precursor symptoms of diseases not generally recognized as “diseases”. Examples of prodromal symptoms of such diseases include mild cognitive impairment (MCI) seen before the onset of Alzheimer's disease.

本発明の組成物によるアミロイド性タンパク質関連疾患の診断は、通常、本発明の組成物を診断対象者又は実験動物などに投与し、その後、脳の画像を撮影し、画像における一般式（Ｉ）又は（II）で表される化合物の状態（量、分布等）に基づいて行う。本発明の組成物の投与方法は特に限定されず、化合物の種類、標識物質の種類などに応じて適宜決めることができるが、通常は、皮内、腹腔内、静脈、動脈、又は脊髄液への注射又は点滴等によって投与する。本発明の組成物の投与量は特に限定されず、化合物の種類、標識物質の種類などに応じて適宜決めることができるが、成人の場合、一般式（Ｉ）又は（II）で表される化合物を１日当たり10^-10〜10^-3mg投与するのが好ましく、10^-8〜10^-5 mg投与するのが更に好ましい。 Diagnosis of amyloid protein-related diseases using the composition of the present invention is usually performed by administering the composition of the present invention to a subject to be diagnosed or a laboratory animal, then taking a brain image, and the general formula (I) in the image. Or based on the state (amount, distribution, etc.) of the compound represented by (II). The administration method of the composition of the present invention is not particularly limited and can be appropriately determined according to the type of compound, the type of labeling substance, etc., but is usually intradermal, intraperitoneal, intravenous, arterial, or spinal fluid. It is administered by injection or infusion. The dose of the composition of the present invention is not particularly limited and can be appropriately determined according to the type of compound, the type of labeling substance, etc. In the case of an adult, it is represented by the general formula (I) or (II) The compound is preferably administered at 10 ⁻¹⁰ to 10 ⁻³ mg per day, more preferably at 10 ⁻⁸ to 10 ⁻⁵ mg.

上記のように本発明の組成物は、通常、注射又は点滴によって投与するので、注射液や点滴液に通常含まれる成分を含んでいてもよい。このような成分としては、液体担体（例えば、リン酸カリウム緩衝液、生理食塩水、リンゲル液、蒸留水、ポリエチレングリコール、植物性油脂、エタノール、グリセリン、ジメチルスルホキサイド、プロピレングリコールなど）、抗菌剤、局所麻酔剤（例えば、塩酸プロカイン、塩酸ジブカインなど）、緩衝液（例えば、トリス−塩酸緩衝液、ヘペス緩衝液など）、浸透圧調節剤（例えば、グルコース、ソルビトール、塩化ナトリウムなど）を例示できる。 As described above, since the composition of the present invention is usually administered by injection or infusion, it may contain components usually contained in injection solutions or infusion solutions. Such components include liquid carriers (for example, potassium phosphate buffer, physiological saline, Ringer's solution, distilled water, polyethylene glycol, vegetable oils, ethanol, glycerin, dimethyl sulfoxide, propylene glycol, etc.), antibacterial agents Local anesthetics (eg, procaine hydrochloride, dibucaine hydrochloride, etc.), buffers (eg, tris-hydrochloric acid buffer, hepes buffer, etc.), osmotic pressure regulators (eg, glucose, sorbitol, sodium chloride, etc.) .

本発明のアミロイド性タンパク質関連疾患診断用組成物は、アミロイド性タンパク質関連疾患の治療薬や予防薬のスクリーニングにも利用できる。例えば、アルツハイマー病などの「疾患」のモデル動物に被験物質を投与した後、前記モデル動物に本発明のアミロイド性タンパク質関連疾患診断用組成物を投与し、その後、前記モデル動物の脳中に含まれる一般式（Ｉ）又は（II）で表される化合物の分布又は量を調べ、その結果、コントロール（被験物質を投与していないモデル動物）との間に有意な差異（例えば、分布部位の縮小、量の減少など）が検出されれば、被験物質はアミロイド性タンパク質関連疾患の治療薬の候補となり得る。また、軽度認知障害などの「疾患の前駆症状」のモデル動物に被験物質を投与した後、前記モデル動物に本発明のアミロイド性タンパク質関連疾患診断用組成物を投与し、その後、前記モデル動物の脳中に含まれる一般式（Ｉ）又は（II）で表される化合物の分布又は量を調べ、その結果、コントロールとの間に有意な差異（例えば、分布部位の縮小又は拡大の鈍化、量の減少又は増大の鈍化など）が検出されれば、被験物質はアミロイド性タンパク質関連疾患の予防薬の候補となり得る。 The composition for diagnosing amyloid protein-related diseases of the present invention can also be used for screening for therapeutic or prophylactic agents for amyloid protein-related diseases. For example, after administering a test substance to a model animal of “disease” such as Alzheimer's disease, the composition for diagnosis of amyloid protein-related disease of the present invention is administered to the model animal, and then contained in the brain of the model animal. The distribution or amount of the compound represented by the general formula (I) or (II) is examined, and as a result, there is a significant difference (for example, distribution site) from the control (model animal not administered with the test substance). If a reduction, a decrease in the amount, etc.) are detected, the test substance can be a candidate for a therapeutic agent for amyloid protein-related diseases. Moreover, after administering a test substance to a model animal of “disease symptom” such as mild cognitive impairment, the composition for amyloid protein-related disease diagnosis of the present invention is administered to the model animal. The distribution or amount of the compound represented by the general formula (I) or (II) contained in the brain is examined, and as a result, there is a significant difference from the control (for example, the reduction or expansion of the distribution site, the amount of reduction, the amount) If a decrease or a decrease in increase is detected, the test substance can be a candidate for a prophylactic agent for amyloid protein-related diseases.

また、本発明のアミロイド性タンパク質関連疾患診断用組成物は、既に効果が確認されているアミロイド性タンパク質関連疾患の治療薬や予防薬の評価にも利用できる。即ち、アミロイド性タンパク質関連疾患のモデル動物に前記疾患の治療薬又は予防薬を投与した後、前記モデル動物に本発明のアミロイド性タンパク質関連疾患診断用組成物を投与し、その後、前記モデル動物の脳中に含まれる一般式（Ｉ）又は（II）で表される化合物の分布又は量を調べ、これにより、前記治療薬や予防薬の評価（具体的には、有効な投与量、有効な投与方法など）を行う。 In addition, the composition for diagnosing amyloid protein-related diseases of the present invention can also be used for the evaluation of therapeutic and prophylactic agents for amyloid protein-related diseases whose effects have already been confirmed. That is, after the therapeutic or prophylactic agent for the disease is administered to a model animal of amyloid protein-related disease, the composition for diagnosing amyloid protein-related disease of the present invention is administered to the model animal, and then The distribution or amount of the compound represented by the general formula (I) or (II) contained in the brain is examined, thereby evaluating the therapeutic agent or the preventive agent (specifically, effective dosage, effective dose). Administration method).

〔実験方法〕
（１）試薬・機器
放射性ヨウ素-125はMP Biomedicals,Inc製Iodine-125 (3.7 GBq/mL)を用いた。逆相HPLCはナカライテスク社製Cosmosil 5C₁₈-AR-IIカラム (4.6×150mm) を用いて、超純水:アセトニトリル=4:6 - 3:7を溶出溶媒として流速1mL/minで分析した。中圧分取クロマトグラフィーは山善社製ULTRA PACK SI-40B カラム (26×300mm)を用いた。¹H NMRはVarian Geminin 300を用い、テトラメチルシランを内標準物質として測定した。質量分析は、JEOL IMS-DX300を用いて測定した。分取TLCはMERCK社製12PLC plates 20×20cm Silica gel 60F₂₅₄, 2mmを用いた。Amyloid β-Protein (Human, 1-42) はペプチド研究所より購入し、その他の試薬は特級試薬を用いた。〔experimental method〕
(1) Reagent / instrument Radioiodine-125 used was Iodine-125 (3.7 GBq / mL) manufactured by MP Biomedicals, Inc. Reverse phase HPLC was performed using a Cosmosil 5C ₁₈ -AR-II column (4.6 × 150 mm) manufactured by Nacalai Tesque with ultrapure water: acetonitrile = 4: 6-3: 7 as an elution solvent at a flow rate of 1 mL / min. For medium pressure preparative chromatography, a ULTRA PACK SI-40B column (26 × 300 mm) manufactured by Yamazen Corporation was used. ¹ H NMR was measured using Varian Geminin 300, using tetramethylsilane as an internal standard substance. Mass spectrometry was measured using JEOL IMS-DX300. Preparative TLC was used MERCK Co. 12PLC plates 20 × 20cm Silica gel 60F 254, 2mm. Amyloid β-Protein (Human, 1-42) was purchased from Peptide Institute, and other reagents were special grades.

（２）2,5-ジフェニル-1,2,4-オキサジアゾール誘導体の合成
3-(4-ブロモフェニル)-5-(4-ニトロフェニル)-1,2,4-オキサジアゾール (化合物1)の合成
4-ブロモベンズアミドオキシム (645 mg, 3 mmol)と4-ニトロ安息香酸 (495 mg, 3 mmol)のDMF (10 mL)溶液に、DCC (3.6 mmol)とHOBT (6.0 mmol)のDMF (5 mL)溶液を加えた。室温で１８時間反応後、100 ℃で２時間加熱還流を行った。反応溶媒を減圧留去後、酢酸エチル／ヘキサン(9/1)を溶出溶媒とするシリカゲルクロマトグラフィに付し、目的とする化合物１を得た（収量370 mg、収率35.6％）。 ¹H NMR (300 MHz, CDCl₃) δ 7.70 (d, J = 8.7 Hz, 2H), 8.06 (d, J = 8.7 Hz, 2H), 8.43 (s, 4H). MS m/z 346 (M⁺). (2) Synthesis of 2,5-diphenyl-1,2,4-oxadiazole derivatives
Synthesis of 3- (4-bromophenyl) -5- (4-nitrophenyl) -1,2,4-oxadiazole (Compound 1)
To a solution of 4-bromobenzamide oxime (645 mg, 3 mmol) and 4-nitrobenzoic acid (495 mg, 3 mmol) in DMF (10 mL), add DCC (3.6 mmol) and HOBT (6.0 mmol) in DMF (5 mL ) The solution was added. After reaction at room temperature for 18 hours, the mixture was heated to reflux at 100 ° C. for 2 hours. After evaporating the reaction solvent under reduced pressure, the residue was subjected to silica gel chromatography using ethyl acetate / hexane (9/1) as an elution solvent to obtain the target compound 1 (yield 370 mg, yield 35.6%). ¹ H NMR (300 MHz, CDCl ₃ ) δ 7.70 (d, J = 8.7 Hz, 2H), 8.06 (d, J = 8.7 Hz, 2H), 8.43 (s, 4H) .MS m / z 346 (M ⁺ ).

4-(3-(4-ブロモフェニル)-1,2,4-オキサジアゾール-5-イル)アニリン (化合物2)の合成
化合物1 (350 mg, 1 mmol)とSnCl₂ (948 mg, 5 mmol)をEtOH (15 mL)に懸濁し、２時間加熱還流した。反応溶液を室温まで戻し、1 M NaOH (100 mL)を加えて、アルカリ状態にした。酢酸エチル(100 mL x 2)で抽出後、生理食塩水で洗浄、硫酸ナトリウムで乾燥後、溶媒を減圧留去することにより、目的とする化合物2を得た（収量258 mg、収率80.8％）。 ¹H NMR (300 MHz, CDCl₃) δ 4.16 (s, 2H), 6.75 (d, J = 8.7 Hz, 2H), 7.63 (d, J = 8.4 Hz, 2H), 8.00 (d, J = 9.0 Hz, 2H), 8.03 (d, J = 6.3 Hz, 2H). MS m/z 316 (M⁺). Synthesis of 4- (3- (4-bromophenyl) -1,2,4-oxadiazol-5-yl) aniline (Compound 2) Compound 1 (350 mg, 1 mmol) and SnCl ₂ (948 mg, 5 mmol) was suspended in EtOH (15 mL) and heated to reflux for 2 hours. The reaction solution was returned to room temperature and made alkaline by adding 1 M NaOH (100 mL). After extraction with ethyl acetate (100 mL x 2), washing with physiological saline and drying over sodium sulfate, the solvent was distilled off under reduced pressure to obtain the target compound 2 (yield 258 mg, yield 80.8%). ). ¹ H NMR (300 MHz, CDCl ₃ ) δ 4.16 (s, 2H), 6.75 (d, J = 8.7 Hz, 2H), 7.63 (d, J = 8.4 Hz, 2H), 8.00 (d, J = 9.0 Hz , 2H), 8.03 (d, J = 6.3 Hz, 2H) .MS m / z 316 (M ⁺ ).

4-(3-(4-ブロモフェニル)-1,2,4-オキサジアゾール-5-イル)-N-メチルアニリン (化合物3)の合成
化合物2 (185 mg, 0.59 mmol)とパラホルムアルデヒド(176 mg, 0.59 mmol)のメタノール溶液(10 mL)に NaOCH₃ (28 wt % in MeOH, 0.4 mL) を滴下した。反応溶液は、３０分間加熱還流し、その後、NaBH₄ (225 mg, 5.9 mmol)を加えて、さらに1.5時間加熱還流した。1 M NaOH (50 mL)を加えて、CHCl₃ (50 mL)で抽出した後、硫酸ナトリウムで乾燥した。残渣は、酢酸エチル／ヘキサン(1/4)を溶出溶媒とするシリカゲルクロマトグラフィに付し、目的とする化合物3を得た（収量98 mgm収率50.3％）。¹H NMR (300 MHz, CDCl₃) δ 2.93 (s, 3H), 4.30 (s, 1H), 6.66 (d, J = 8.7 Hz, 2H), 7.63 (d, J = 8.7 Hz, 2H), 8.01 (d, J = 8.7 Hz, 2H), 8.03 (d, J = 8.7 Hz, 2H). MS m/z 330 (M⁺). Synthesis of 4- (3- (4-bromophenyl) -1,2,4-oxadiazol-5-yl) -N-methylaniline (Compound 3) Compound 2 (185 mg, 0.59 mmol) and paraformaldehyde ( NaOCH ₃ (28 wt% in MeOH, 0.4 mL) was added dropwise to a methanol solution (10 mL) of 176 mg, 0.59 mmol). The reaction solution was heated to reflux for 30 minutes, then NaBH ₄ (225 mg, 5.9 mmol) was added, and the mixture was further heated to reflux for 1.5 hours. 1 M NaOH (50 mL) was added, and the mixture was extracted with CHCl ₃ (50 mL) and dried over sodium sulfate. The residue was subjected to silica gel chromatography using ethyl acetate / hexane (1/4) as an elution solvent to obtain the target compound 3 (yield 98 mgm, 50.3%). ¹ H NMR (300 MHz, CDCl ₃ ) δ 2.93 (s, 3H), 4.30 (s, 1H), 6.66 (d, J = 8.7 Hz, 2H), 7.63 (d, J = 8.7 Hz, 2H), 8.01 (d, J = 8.7 Hz, 2H), 8.03 (d, J = 8.7 Hz, 2H). MS m / z 330 (M ⁺ ).

4-(3-(4-ブロモフェニル)-1,2,4-オキサジアゾール-5-イル)-N,N-ジメチルアニリン (化合物4)の合成
化合物2 (35 mg, 0.10 mmol) とパラホルムアルデヒド(36 mg, 1.2 mmol)を酢酸(5 mL)溶液に、NaCNBH₃ (50 mg, 0.80 mmol)を加えた。室温で２時間反応を行い、1 M NaOH (30 mL)を加えた後、CH₃Cl (30 mL)で抽出した。硫酸ナトリウムで乾燥後、酢酸エチル／ヘキサン(1/4)を溶出溶媒とするシリカゲルクロマトグラフィに付し、目的とする化合物4を得た（収量24 mg、収率68.4％）。¹H NMR (300 MHz, CDCl₃) δ 3.09 (s, 6H), 6.75 (d, J = 9.0 Hz, 2H), 7.63 (d, J = 8.4 Hz, 2H), 8.35 (d, J = 8.7 Hz, 2H), 8.43 (d, J = 8.7 Hz, 2H). MS m/z 344. Synthesis of 4- (3- (4-bromophenyl) -1,2,4-oxadiazol-5-yl) -N, N-dimethylaniline (Compound 4) Compound 2 (35 mg, 0.10 mmol) and para To a solution of formaldehyde (36 mg, 1.2 mmol) in acetic acid (5 mL) was added NaCNBH ₃ (50 mg, 0.80 mmol). The reaction was performed at room temperature for 2 hours, 1 M NaOH (30 mL) was added, and the mixture was extracted with CH ₃ Cl (30 mL). After drying over sodium sulfate, the product was subjected to silica gel chromatography using ethyl acetate / hexane (1/4) as an elution solvent to obtain the target compound 4 (yield 24 mg, yield 68.4%). ¹ H NMR (300 MHz, CDCl ₃ ) δ 3.09 (s, 6H), 6.75 (d, J = 9.0 Hz, 2H), 7.63 (d, J = 8.4 Hz, 2H), 8.35 (d, J = 8.7 Hz , 2H), 8.43 (d, J = 8.7 Hz, 2H) .MS m / z 344.

3-(4-ブロモフェニル)-5-(4-メトキシフェニル)-1,2,4-オキサジアゾール (化合物5)の合成
化合物 1 を得る際に用いた方法により、目的とする化合物5を得た（収量153 mg、収率23.1％）。¹H NMR (300 MHz, CDCl₃) δ 3.91 (s, 3H), 7.05 (d, J = 8.7 Hz, 2H), 7.65 (d, J = 8.7 Hz, 2H), 8.04 (d, J = 8.4 Hz, 2H), 8.15 (d, J = 9.0 Hz, 2H). MS m/z 330 (M⁺). Synthesis of 3- (4-bromophenyl) -5- (4-methoxyphenyl) -1,2,4-oxadiazole (Compound 5) According to the method used to obtain Compound 1, the target compound 5 was synthesized. Obtained (yield 153 mg, yield 23.1%). ¹ H NMR (300 MHz, CDCl ₃ ) δ 3.91 (s, 3H), 7.05 (d, J = 8.7 Hz, 2H), 7.65 (d, J = 8.7 Hz, 2H), 8.04 (d, J = 8.4 Hz , 2H), 8.15 (d, J = 9.0 Hz, 2H) .MS m / z 330 (M ⁺ ).

4-(3-(4-ブロモフェニル)-1,2,4-オキサジアゾール-5-イル)フェノール (化合物6)の合成
氷冷下、化合物5 (300 mg, 0.91 mmol)のCH₂Cl₂ (10 mL)溶液に、BBr₃ (4.5 mL, 1 M solution in CH₂Cl₂)を滴下した。その後４２時間室温で反応を行い、水 (30 mL)を加えて反応を終了した。クロロホルム (30 mL x 2)で抽出を行い、有機層を硫酸ナトリウムで乾燥し、残渣をヘキサン／酢酸エチル（4/1）を溶出溶媒とするシリカゲルクロマトグラフィに付し、目的とする化合物6を得た（収量146 mg、収率50.6％）。¹H NMR (300 MHz, CDCl₃) d 6.99 (d, J = 8.7 Hz, 2H), 7.65 (d, J = 8.7 Hz, 2H), 8.04 (d, J = 8.1 Hz, 2H), 8.12 (d, J = 9.0 Hz, 2H). MS m/z 316 (M⁺). Synthesis of 4- (3- (4-bromophenyl) -1,2,4-oxadiazol-5-yl) phenol (Compound 6) Under cooling with ice, Compound 5 (300 mg, 0.91 mmol) in CH ₂ Cl BBr ₃ (4.5 mL, 1 M solution in CH ₂ Cl ₂ ) was added dropwise to the ₂ (10 mL) solution. Thereafter, the reaction was performed at room temperature for 42 hours, and water (30 mL) was added to terminate the reaction. Extraction was performed with chloroform (30 mL x 2), the organic layer was dried over sodium sulfate, and the residue was subjected to silica gel chromatography using hexane / ethyl acetate (4/1) as an elution solvent to obtain the target compound 6. (Yield 146 mg, Yield 50.6%). ¹ H NMR (300 MHz, CDCl ₃ ) d 6.99 (d, J = 8.7 Hz, 2H), 7.65 (d, J = 8.7 Hz, 2H), 8.04 (d, J = 8.1 Hz, 2H), 8.12 (d , J = 9.0 Hz, 2H) .MS m / z 316 (M ⁺ ).

4-(3-(4-(トリブチルスタンニル)フェニル)-1,2,4-オキサジアゾール-5-イル)アニリン (化合物7)の合成
化合物2 (100 mg, 0.32 mmol), ビス(トリブチルスズ) (0.2 mL)および(Ph₃P)₄Pd (16 mg, 0.014 mmol)をジオキサンとトリエチルアミンの混合溶媒(10 mL, 3:2 dioxane/triethylamine mixture)に溶解し、１０時間加熱還流を行った。反応溶媒を留去し、残渣をヘキサン／酢酸エチル（３／１）を溶出溶媒とするシリカゲルクロマトグラフィに付し、目的とする化合物7を得た（収量28 mg、収率16.8％）。¹H NMR (300 MHz, CDCl₃) δ 0.87-1.61 (m, 27H), 4.13 (s, 2H), 6.76 (d, J = 8.7 Hz, 2H), 7.59 (d, J = 8.1 Hz, 2H), 8.01 (d, J = 8.7 Hz, 1H), 8.07 (d, J = 8.1 Hz, 2H), 8.28 (s, 1H). Synthesis of 4- (3- (4- (tributylstannyl) phenyl) -1,2,4-oxadiazol-5-yl) aniline (Compound 7) Compound 2 (100 mg, 0.32 mmol), bis (tributyltin ) (0.2 mL) and (Ph ₃ P) ₄ Pd (16 mg, 0.014 mmol) were dissolved in a mixed solvent of dioxane and triethylamine (10 mL, 3: 2 dioxane / triethylamine mixture) and heated to reflux for 10 hours. . The reaction solvent was distilled off, and the residue was subjected to silica gel chromatography using hexane / ethyl acetate (3/1) as an elution solvent to obtain the target compound 7 (yield 28 mg, yield 16.8%). ¹ H NMR (300 MHz, CDCl ₃ ) δ 0.87-1.61 (m, 27H), 4.13 (s, 2H), 6.76 (d, J = 8.7 Hz, 2H), 7.59 (d, J = 8.1 Hz, 2H) , 8.01 (d, J = 8.7 Hz, 1H), 8.07 (d, J = 8.1 Hz, 2H), 8.28 (s, 1H).

N-メチル-4-(3-(4-(トリブチルスタンニル)フェニル)-1,2,4-オキサジアゾール-5-イル)アニリン (化合物8)の合成
化合物7を得るために用いた方法により、目的とする化合物8を得た（収量23 mg、収率15.8%）。¹H NMR (300 MHz, CDCl₃) δ 0.87-1.63 (m, 27H), 2.92 (s, 3H), 4.27 (s, 1H), 6.66 (d, J = 8.7 Hz, 2H), 7.59 (d, J = 8.1 Hz, 2H), 8.03 (d, J = 8.4 Hz, 2H), 8.08 (d, J = 7.8 Hz, 2H). Synthesis of N-methyl-4- (3- (4- (tributylstannyl) phenyl) -1,2,4-oxadiazol-5-yl) aniline (Compound 8) Method used to obtain Compound 7 Gave the target compound 8 (yield 23 mg, yield 15.8%). ¹ H NMR (300 MHz, CDCl ₃ ) δ 0.87-1.63 (m, 27H), 2.92 (s, 3H), 4.27 (s, 1H), 6.66 (d, J = 8.7 Hz, 2H), 7.59 (d, J = 8.1 Hz, 2H), 8.03 (d, J = 8.4 Hz, 2H), 8.08 (d, J = 7.8 Hz, 2H).

N,N-ジメチル-4-(3-(4-(トリブチルスタンニル)フェニル)-1,2,4-オキサジアゾール-5-イル)アニリン (化合物9)の合成
化合物7 を得る際に用いた方法により、目的とする化合物9 を得た（収量45 mg、収率20.3％）。¹H NMR (300 MHz, CDCl₃) δ 0.87-1.58 (m, 27H), 3.09 (s, 6H), 6.76 (d, J = 9.6 Hz, 2H), 7.59 (d, J = 8.1 Hz, 2H), 8.05 (d, J = 8.4 Hz, 2H), 8.08 (d, J = 8.4 Hz, 2H). Synthesis of N, N-dimethyl-4- (3- (4- (tributylstannyl) phenyl) -1,2,4-oxadiazol-5-yl) aniline (Compound 9) Used to obtain Compound 7. The target compound 9 was obtained by the conventional method (yield 45 mg, yield 20.3%). ¹ H NMR (300 MHz, CDCl ₃ ) δ 0.87-1.58 (m, 27H), 3.09 (s, 6H), 6.76 (d, J = 9.6 Hz, 2H), 7.59 (d, J = 8.1 Hz, 2H) , 8.05 (d, J = 8.4 Hz, 2H), 8.08 (d, J = 8.4 Hz, 2H).

5-(4-メトキシフェニル)-3-(4-(トリブチルスタンニル)フェニル)-1,2,4-オキサジアゾール (化合物10)の合成
化合物7を合成した方法を用いて、目的とする化合物10を得た（収量42 mg、収率22.8％）。¹H NMR (300 MHz, CDCl₃) δ 0.87-1.59 (m, 27H), 3.91 (s, 3H), 7.04 (d, J = 8.7 Hz, 2H), 7.61 (d, J = 7.8 Hz, 2H), 8.07 (d, J = 9.0 Hz, 2H), 8.17 (d, J = 9.0 Hz, 2H). Synthesis of 5- (4-methoxyphenyl) -3- (4- (tributylstannyl) phenyl) -1,2,4-oxadiazole (Compound 10) Compound 10 was obtained (yield 42 mg, yield 22.8%). ¹ H NMR (300 MHz, CDCl ₃ ) δ 0.87-1.59 (m, 27H), 3.91 (s, 3H), 7.04 (d, J = 8.7 Hz, 2H), 7.61 (d, J = 7.8 Hz, 2H) , 8.07 (d, J = 9.0 Hz, 2H), 8.17 (d, J = 9.0 Hz, 2H).

4-(3-(4-(トリブチルスタンニル)フェニル)-1,2,4-オキサジアゾール-5-イル)フェノール (化合物11)の合成
化合物 7 を合成した方法を用いて、目的とする化合物11を得た（収量28 mg、収率60.1％）。¹H NMR (300 MHz, CDCl₃) δ 0.87-1.58 (m, 27H), 6.99 (d, J = 9.0 Hz, 2H), 7.61 (d, J = 7.8 Hz, 2H), 8.07 (d, J = 8.7 Hz, 2H), 8.12 (d, J = 8.7 Hz, 2H). Synthesis of 4- (3- (4- (tributylstannyl) phenyl) -1,2,4-oxadiazol-5-yl) phenol (Compound 11) Compound 11 was obtained (yield 28 mg, yield 60.1%). ¹ H NMR (300 MHz, CDCl ₃ ) δ 0.87-1.58 (m, 27H), 6.99 (d, J = 9.0 Hz, 2H), 7.61 (d, J = 7.8 Hz, 2H), 8.07 (d, J = 8.7 Hz, 2H), 8.12 (d, J = 8.7 Hz, 2H).

4-(3-(4-ヨードフェニル)-1,2,4-オキサジアゾール-5-イル)アニリン (化合物12)の合成
化合物7 (27 mg, 0.05 mmol)のCHCl₃ (5 mL) 溶液に、ヨウ素のクロロホルム溶液(1 mL, 50 mg/mL)を加えた。室温で１０分間反応を行い、飽和亜硫酸水素ナトリウム水溶液 (25 mL)を加えることにより、反応を終了した。クロロホルム (25 mL x 2)で抽出した後、有機層を硫酸ナトリウムで乾燥し、溶媒を減圧留去した。残渣をヘキサンで洗浄することによって、目的とする化合物12を得た（収量12 mg、収率66.1％）。¹H NMR (300 MHz, CDCl₃) δ 4.15 (s, 2H), 6.76 (d, J = 8.7 Hz, 2H), 7.83 (d, J = 8.4 Hz, 2H), 7.89 (d, J = 8.4 Hz, 2H), 8.01 (d, J = 8.7 Hz, 2H). HRMS m/z C₁₄H₁₀N₃OI found 362.9855 / calcld 362.9869 (M⁺). Synthesis of 4- (3- (4-iodophenyl) -1,2,4-oxadiazol-5-yl) aniline (Compound 12) Compound 7 (27 mg, 0.05 mmol) in CHCl ₃ (5 mL) Was added a solution of iodine in chloroform (1 mL, 50 mg / mL). The reaction was performed at room temperature for 10 minutes, and the reaction was terminated by adding a saturated aqueous sodium hydrogen sulfite solution (25 mL). After extraction with chloroform (25 mL × 2), the organic layer was dried over sodium sulfate, and the solvent was distilled off under reduced pressure. The residue was washed with hexane to obtain the target compound 12 (yield 12 mg, yield 66.1%). ¹ H NMR (300 MHz, CDCl ₃ ) δ 4.15 (s, 2H), 6.76 (d, J = 8.7 Hz, 2H), 7.83 (d, J = 8.4 Hz, 2H), 7.89 (d, J = 8.4 Hz , 2H), 8.01 (d, J = 8.7 Hz, 2H) .HRMS m / z C ₁₄ H ₁₀ N ₃ OI found 362.9855 / calcld 362.9869 (M ⁺ ).

4-(3-(4-ヨードフェニル)-1,2,4-オキサジアゾール-5-イル)-N-メチルアニリン (化合物13)の合成
化合物8を出発原料とし、化合物12の合成方法と同様のヨウ素化反応により、目的とする化合物13を得た（収量20 mg、収率86.0％）。¹H NMR (300 MHz, CDCl₃) δ 2.93 (s, 3H), 4.29 (s, 1H), 6.66 (d, J = 8.4 Hz, 2H), 7.85 (d, J = 8.4 Hz, 2H), 7.88 (d, J = 8.4 Hz, 2H), 8.02 (d, J = 8.4 Hz, 2H). HRMS m/z C₁₅H₁₂N₃OI found 377.0022 / calcld 377.0025 (M⁺). Synthesis of 4- (3- (4-iodophenyl) -1,2,4-oxadiazol-5-yl) -N-methylaniline (Compound 13) Using Compound 8 as a starting material, The target compound 13 was obtained by the same iodination reaction (yield 20 mg, yield 86.0%). ¹ H NMR (300 MHz, CDCl ₃ ) δ 2.93 (s, 3H), 4.29 (s, 1H), 6.66 (d, J = 8.4 Hz, 2H), 7.85 (d, J = 8.4 Hz, 2H), 7.88 (d, J = 8.4 Hz, 2H), 8.02 (d, J = 8.4 Hz, 2H). HRMS m / z C ₁₅ H ₁₂ N ₃ OI found 377.0022 / calcld 377.0025 (M ⁺ ).

4-(3-(4-ヨードフェニル)-1,2,4-オキサジアゾール-5-イル)-N,N-ジメチルアニリン (化合物14)の合成
化合物9を出発原料とし、化合物12の合成方法と同様のヨウ素化反応により、目的とする化合物14を得た（収量34 mg、収率72.4％）。¹H NMR (300 MHz, CDCl₃) δ 3.09 (s, 6H), 6.75 (d, J = 9.3 Hz, 2H), 7.83 (d, J = 8.7 Hz, 2H), 7.90 (d, J = 8.7 Hz, 2H), 8.04 (d, J = 9.3 Hz, 2H). HRMS m/z C₁₆H₁₄N₃OI found 391.0192 / calcld 391.0182 (M⁺). Synthesis of 4- (3- (4-iodophenyl) -1,2,4-oxadiazol-5-yl) -N, N-dimethylaniline (Compound 14) Synthesis of Compound 12 using Compound 9 as a starting material The target compound 14 was obtained by the iodination reaction similar to the method (yield 34 mg, yield 72.4%). ¹ H NMR (300 MHz, CDCl ₃ ) δ 3.09 (s, 6H), 6.75 (d, J = 9.3 Hz, 2H), 7.83 (d, J = 8.7 Hz, 2H), 7.90 (d, J = 8.7 Hz , 2H), 8.04 (d, J = 9.3 Hz, 2H) .HRMS m / z C ₁₆ H ₁₄ N ₃ OI found 391.0192 / calcld 391.0182 (M ⁺ ).

3-(4-ヨードフェニル)-5-(4-メトキシフェニル)-1,2,4-オキサジアゾール (化合物15)の合成
化合物10を出発原料とし、化合物12の合成方法と同様のヨウ素化反応により、目的とする化合物15を得た（収量17 mg、収率69.2％）。¹H NMR (300 MHz, CDCl₃) δ 3.89 (s, 3H), 6.97 (d, J = 9.3 Hz, 2H), 7.48 (d, J = 8.4 Hz, 2H), 7.80 (d, J = 9.3 Hz, 2H), 7.85 (d, J = 9.0 Hz, 2H). HRMS m/z C₁₅H₁₁N₂O₂I found 377.9865 / calcld 377.9872 (M⁺). Synthesis of 3- (4-iodophenyl) -5- (4-methoxyphenyl) -1,2,4-oxadiazole (Compound 15) Iodination similar to the synthesis method of Compound 12 using Compound 10 as a starting material The target compound 15 was obtained by the reaction (yield 17 mg, yield 69.2%). ¹ H NMR (300 MHz, CDCl ₃ ) δ 3.89 (s, 3H), 6.97 (d, J = 9.3 Hz, 2H), 7.48 (d, J = 8.4 Hz, 2H), 7.80 (d, J = 9.3 Hz , 2H), 7.85 (d, J = 9.0 Hz, 2H) .HRMS m / z C ₁₅ H ₁₁ N ₂ O ₂ I found 377.9865 / calcld 377.9872 (M ⁺ ).

4-(3-(4-ヨードフェニル)-1,2,4-オキサジアゾール-5-イル)フェノール (化合物16)の合成
化合物11を出発原料とし、化合物12の合成方法と同様のヨウ素化反応により、目的とする化合物16を得た（収量14 mg、収率81.8％）。 ¹H NMR (300 MHz, CDCl₃) δ 6.99 (d, J = 8.7 Hz, 2H), 7.87 (d, J = 8.4 Hz, 2H), 7.88 (d, J = 8.1 Hz, 2H), 8.12 (d, J = 8.1 Hz, 2H). HRMS m/z C₁₄H₉N₂O₂I found 363.9704 / calcld 363.9709 (M⁺). Synthesis of 4- (3- (4-iodophenyl) -1,2,4-oxadiazol-5-yl) phenol (Compound 16) Iodination similar to the synthesis method of Compound 12 using Compound 11 as a starting material The target compound 16 was obtained by the reaction (yield 14 mg, yield 81.8%). ¹ H NMR (300 MHz, CDCl ₃ ) δ 6.99 (d, J = 8.7 Hz, 2H), 7.87 (d, J = 8.4 Hz, 2H), 7.88 (d, J = 8.1 Hz, 2H), 8.12 (d , J = 8.1 Hz, 2H) .HRMS m / z C ₁₄ H ₉ N ₂ O ₂ I found 363.9704 / calcld 363.9709 (M ⁺ ).

（２）2,5-ジフェニル-1,3,4-オキサジアゾール誘導体の合成
4-(5-(4-ブロモフェニル)-1,3,4-オキサジアゾール-2-イル)-N,N-ジメチルベンゼンアミン (化合物17)の合成
4-ブロモ安息香酸ヒドラジド(215 mg, 1 mmol)と4-ジメチルアミノベンズアルデヒド(149 mg, 1 mmol)、硝酸セリウムアンモニウム(548 mg, 1 mmol) をジクロロメタン (10 mL) に溶解し24時間加熱還流した。精製水を加えてクロロホルムで抽出し、無水硫酸ナトリウムで乾燥後、溶媒を減圧留去した。残渣を酢酸エチル/ヘキサン (1/4) を溶出溶媒とする中圧分取クロマトグラフィーに付し、化合物17を得た。収量12 mg (収率3.5％)¹H NMR (300 MHz, CDCl₃) δ3.08 (s, 6H), 6.76 (d, J=9.0 Hz, 2H), 7.66 (d, J=8.4 Hz, 2H), 7.98 (dd, J=5.4, 4.5 Hz, 4H). MS m/z 362 (M⁺). (2) Synthesis of 2,5-diphenyl-1,3,4-oxadiazole derivatives
Synthesis of 4- (5- (4-bromophenyl) -1,3,4-oxadiazol-2-yl) -N, N-dimethylbenzenamine (Compound 17)
4-Bromobenzoic acid hydrazide (215 mg, 1 mmol), 4-dimethylaminobenzaldehyde (149 mg, 1 mmol) and ceric ammonium nitrate (548 mg, 1 mmol) are dissolved in dichloromethane (10 mL) and heated to reflux for 24 hours. did. Purified water was added, the mixture was extracted with chloroform, dried over anhydrous sodium sulfate, and the solvent was evaporated under reduced pressure. The residue was subjected to medium pressure preparative chromatography using ethyl acetate / hexane (1/4) as an elution solvent to obtain Compound 17. Yield 12 mg (Yield 3.5%) ¹ H NMR (300 MHz, CDCl ₃ ) δ3.08 (s, 6H), 6.76 (d, J = 9.0 Hz, 2H), 7.66 (d, J = 8.4 Hz, 2H ), 7.98 (dd, J = 5.4, 4.5 Hz, 4H) .MS m / z 362 (M ⁺ ).

4-(5-(4-トリブチルスタンニル)フェニル)-1,3,4-オキサジアゾール-2-イル)-N,N-ジメチルベンゼンアミン (化合物18)の合成
化合物17 (19 mg, 0.06 mmol)を1,4-ジオキサン (3 mL) に溶解し、ビス(トリブチルスズ) (0.04 mL)、テトラトリフェニルホスフィンパラジウム (3 mg, 0.002 mmol)、トリエチルアミン (3 mL) を加えて4時間30分、加熱還流した。反応溶媒を減圧留去し、残渣を酢酸エチル/ヘキサン(1/3)を展開溶媒とする分取用TLCにより精製を行い、化合物18を得た。収量2.5mg (収率8.2％) ¹H NMR (300 MHz, CDCl₃) δ0.87-1.6 (m, 27H) 3.07 (s, 6H), 6.77 (d, J=9.0 Hz, 2H), 7.61 (d, J=8.4 Hz, 2H), 8.01 (dd, J=9.0, 8.1 Hz, 4H). Synthesis of 4- (5- (4-tributylstannyl) phenyl) -1,3,4-oxadiazol-2-yl) -N, N-dimethylbenzenamine (Compound 18)
Compound 17 (19 mg, 0.06 mmol) dissolved in 1,4-dioxane (3 mL), bis (tributyltin) (0.04 mL), tetratriphenylphosphine palladium (3 mg, 0.002 mmol), triethylamine (3 mL) And heated to reflux for 4 hours 30 minutes. The reaction solvent was distilled off under reduced pressure, and the residue was purified by preparative TLC using ethyl acetate / hexane (1/3) as a developing solvent to obtain Compound 18. Yield 2.5 mg (8.2% yield) ¹ H NMR (300 MHz, CDCl ₃ ) δ0.87-1.6 (m, 27H) 3.07 (s, 6H), 6.77 (d, J = 9.0 Hz, 2H), 7.61 ( d, J = 8.4 Hz, 2H), 8.01 (dd, J = 9.0, 8.1 Hz, 4H).

4-(5-(ヨードフェニル)-1,3,4-オキサジアゾール-2-イル)-N,N-ジメチルベンゼンアミン (化合物19)の合成
4-ヨード安息香酸ヒドラジド(524 mg, 2 mmol)と4-ジメチルアミノベンズアルデヒド(298 mg, 2 mmol)、硝酸セリウムアンモニウム(1096 mg, 2 mmol) をジクロロメタン (20 mL) に溶解し11時間加熱還流した。精製水を加えてクロロホルムで抽出し、無水硫酸ナトリウムで乾燥後、溶媒を減圧留去した。残渣をクロロホルム/メタノール (99/1) を展開溶媒とする分取用TLCにより精製を行い、化合物19を得た。収量6 mg (収率0.76％)¹H NMR (300 MHz, CDCl₃) δ3.07 (s, 6H), 6.76 (d, J=3.0 Hz, 2H), 7.85 (d, J=12.0 Hz, 4H), 7.97 (d, J=3.0 Hz, 2H). MS m/z 391 (M⁺). Synthesis of 4- (5- (iodophenyl) -1,3,4-oxadiazol-2-yl) -N, N-dimethylbenzenamine (Compound 19)
4-Iodobenzoic acid hydrazide (524 mg, 2 mmol), 4-dimethylaminobenzaldehyde (298 mg, 2 mmol) and ceric ammonium nitrate (1096 mg, 2 mmol) are dissolved in dichloromethane (20 mL) and heated to reflux for 11 hours. did. Purified water was added, the mixture was extracted with chloroform, dried over anhydrous sodium sulfate, and the solvent was evaporated under reduced pressure. The residue was purified by preparative TLC using chloroform / methanol (99/1) as a developing solvent to obtain Compound 19. Yield 6 mg (Yield 0.76%) ¹ H NMR (300 MHz, CDCl ₃ ) δ3.07 (s, 6H), 6.76 (d, J = 3.0 Hz, 2H), 7.85 (d, J = 12.0 Hz, 4H ), 7.97 (d, J = 3.0 Hz, 2H) .MS m / z 391 (M ⁺ ).

2-(4-ヨードフェニル)-5-(4-メトキシフェニル)-1,3,4-オキサジアゾール (化合物20)の合成
4-ヨード安息香酸ヒドラジド(524 mg, 2 mmol)と4-メトキシベンズアルデヒド(272 mg, 2 mmol)、硝酸セリウムアンモニウム (1096 mg, 2 mmol) をジクロロメタン (20mL) に溶解し19時間加熱還流した。精製水を加えてクロロホルムで抽出し、無水硫酸ナトリウムで乾燥後、溶媒を減圧留去した。残渣をクロロホルム/メタノール (99/1) を展開溶媒とする分取用TLCにより精製を行い、化合物20を得た。収量40mg (収率8.8％)¹H NMR (300 MHz, CDCl₃) δ3.89 (s, 3H), 7.03 (d, J=2.9 Hz, 2H), 7.86 (q, J=7.8 Hz, 4H), 8.03 (d, J=3.0 Hz, 2H). MS m/z 378 (M⁺). Synthesis of 2- (4-iodophenyl) -5- (4-methoxyphenyl) -1,3,4-oxadiazole (Compound 20)
4-Iodobenzoic acid hydrazide (524 mg, 2 mmol), 4-methoxybenzaldehyde (272 mg, 2 mmol) and ceric ammonium nitrate (1096 mg, 2 mmol) were dissolved in dichloromethane (20 mL) and heated to reflux for 19 hours. Purified water was added, the mixture was extracted with chloroform, dried over anhydrous sodium sulfate, and the solvent was evaporated under reduced pressure. The residue was purified by preparative TLC using chloroform / methanol (99/1) as a developing solvent to obtain Compound 20. Yield 40 mg (Yield 8.8%) ¹ H NMR (300 MHz, CDCl ₃ ) δ 3.89 (s, 3H), 7.03 (d, J = 2.9 Hz, 2H), 7.86 (q, J = 7.8 Hz, 4H) , 8.03 (d, J = 3.0 Hz, 2H). MS m / z 378 (M ⁺ ).

2-(4-(トリブチルスタンニル)フェニル)-5-(4-メトキシフェニル)-1,3,4-オキサジアゾール (化合物21)の合成
化合物20 (64 mg, 0.06 mmol) を1,4-ジオキサン (5 mL) に溶解し、ビス(トリブチルスズ) (0.11 mL)、テトラトリフェニルホスフィンパラジウム (8.1 mg, mmol)、トリエチルアミン (5 mL) を加えて4時間加熱還流した。反応溶媒を減圧留去し、残渣を酢酸エチル/ヘキサン (1/4) を展開溶媒とする分取用TLCにより精製を行い、化合物21を得た。収量6 mg (収率 6.5％) ¹H NMR (300 MHz, CDCl₃) δ0.87-1.58 (m, 27H), 3.91 (s, 3H), 7.04 (d, J=3.1 Hz, 2H), 7.63 (d, J=2.6 Hz, 2H), 8.06 (q, J=6.6 Hz, 4H). Synthesis of 2- (4- (tributylstannyl) phenyl) -5- (4-methoxyphenyl) -1,3,4-oxadiazole (Compound 21) Compound 20 (64 mg, 0.06 mmol) was converted to 1,4 -Dissolved in dioxane (5 mL), bis (tributyltin) (0.11 mL), tetratriphenylphosphine palladium (8.1 mg, mmol) and triethylamine (5 mL) were added, and the mixture was heated to reflux for 4 hours. The reaction solvent was distilled off under reduced pressure, and the residue was purified by preparative TLC using ethyl acetate / hexane (1/4) as a developing solvent to obtain Compound 21. Yield 6 mg (Yield 6.5%) ¹ H NMR (300 MHz, CDCl ₃ ) δ0.87-1.58 (m, 27H), 3.91 (s, 3H), 7.04 (d, J = 3.1 Hz, 2H), 7.63 (d, J = 2.6 Hz, 2H), 8.06 (q, J = 6.6 Hz, 4H).

4-(5-(4-ヨードフェニル)-1,3,4-オキサジアゾール-2-イル)フェノール (化合物22)の合成
化合物21 (36mg, 0.1 mmol) をジクロロメタン溶液に溶解し、氷冷下三臭化ホウ素ジクロロメタン溶液 (0.6mL) を徐々に加えていった。室温で5日間反応させた後、反応液に少量ずつ精製水を加え反応を停止させた。クロロホルムで抽出後、水層を酢酸エチルで抽出し、無水硫酸ナトリウムで乾燥後、溶媒を減圧留去した。残渣を酢酸エチル/ヘキサン (1/2) を溶出溶媒とする中圧分取クロマトグラフィーに付し、化合物22を得た。収量17 mg (収率 49％)¹H NMR (300 MHz, CDCl₃) δ6.98-7.06 (m, 2H), 7.86-7.91 (m, 4H), 8.02-8.09 (m, 2H). MS m/z 364 (M⁺). Synthesis of 4- (5- (4-iodophenyl) -1,3,4-oxadiazol-2-yl) phenol (Compound 22) Compound 21 (36 mg, 0.1 mmol) was dissolved in a dichloromethane solution and ice-cooled. Lower boron tribromide dichloromethane solution (0.6 mL) was added slowly. After reacting at room temperature for 5 days, purified water was added to the reaction solution little by little to stop the reaction. After extraction with chloroform, the aqueous layer was extracted with ethyl acetate, dried over anhydrous sodium sulfate, and the solvent was evaporated under reduced pressure. The residue was subjected to medium pressure preparative chromatography using ethyl acetate / hexane (1/2) as an elution solvent to obtain Compound 22. Yield 17 mg (Yield 49%) ¹ H NMR (300 MHz, CDCl ₃ ) δ6.98-7.06 (m, 2H), 7.86-7.91 (m, 4H), 8.02-8.09 (m, 2H). MS m / z 364 (M ⁺ ).

（３）DPOD誘導体の ¹²⁵ I標識
種々のトリブチルスズ化合物 (1 mg/mL) のエタノール溶液 (50 μL) に[¹²⁵I]NaI (1-5 μCi)、1 N塩酸 (50 μL) を加え、最後に3% w/v 過酸化水素水 (50 μL) を加えた。2分間室温放置後、飽和亜硫酸水素ナトリウム水溶液 (100 μL) を加え反応を停止させ、飽和炭酸水素ナトリウム水溶液 (100 μL) を加えて反応溶液を中和した。酢酸エチルで抽出し、硫酸ナトリウムを入れたパスツールピペットに通して脱水した後、逆相HPLC (水 : アセトニトリル = 4 : 6) で精製した。非放射性化合物を標品として254 nmにおける吸光度をHPLCで分析し、それと一致する目的物を分取し、酢酸エチルで抽出し、窒素気流下酢酸エチルを留去した。 (3) ¹²⁵ I-labeled DPOD derivative [ ¹²⁵ I] NaI (1-5 μCi) and 1 N hydrochloric acid (50 μL) were added to ethanol solutions (50 μL) of various tributyltin compounds (1 mg / mL). 3% w / v hydrogen peroxide solution (50 μL) was added. After standing at room temperature for 2 minutes, a saturated aqueous sodium hydrogen sulfite solution (100 μL) was added to stop the reaction, and a saturated aqueous sodium hydrogen carbonate solution (100 μL) was added to neutralize the reaction solution. The mixture was extracted with ethyl acetate, dehydrated through a Pasteur pipette containing sodium sulfate, and purified by reverse phase HPLC (water: acetonitrile = 4: 6). Absorbance at 254 nm was analyzed by HPLC using a non-radioactive compound as a standard product, and the target product corresponding to the sample was collected, extracted with ethyl acetate, and ethyl acetate was distilled off under a nitrogen stream.

（４）Ab(1-42)凝集体の調製
Ab(1-42)を、1 mM EDTAを含んだ10 mM リン酸緩衝液 (pH 7.4) に0.25 mg/mLの濃度になるよう溶解し、37 ℃で42時間インキュベートすることにより、Ab(1-42)凝集体を調製した。 (4) Preparation of Ab (1-42) aggregates
Ab (1-42) was dissolved in 10 mM phosphate buffer (pH 7.4) containing 1 mM EDTA to a concentration of 0.25 mg / mL, and incubated at 37 ° C for 42 hours to obtain Ab (1 -42) Aggregates were prepared.

（５）Ab(1-42)凝集体を用いた結合実験
10％EtOH溶液 900 μL、種々の濃度のサンプル溶液50 μL、Aβ (1-42)凝集体50 μLまたはリン酸Buffer 50 μLを加え、室温で3時間放置した。Aβ(1-42)凝集体と結合した化合物と結合していない化合物とは、Whatman GF/B filterを用いてBrandel M-24R cell harvester によって分離し、濾過したフィルターに残った放射能をγカウンターで計測した。 (5) Binding experiments using Ab (1-42) aggregates
900 μL of 10% EtOH solution, 50 μL of sample solutions of various concentrations, 50 μL of Aβ (1-42) aggregates or 50 μL of phosphate buffer were added and left at room temperature for 3 hours. The compound bound to the Aβ (1-42) aggregate and the unbound compound are separated by Brandel M-24R cell harvester using Whatman GF / B filter, and the radioactivity remaining on the filtered filter is determined by the γ counter. Measured with.

（６）Ab(1-42)凝集体を用いた結合阻害実験による阻害定数：K _i 値の算出
10%EtOH溶液 850 μL、[¹²⁵I]IMPYの10％エタノール溶液 50 μL、種々の濃度のサンプル溶液 50 μL (0〜20 μM) を混和し、最後にAb(1-42)凝集体溶液 50 μLを加え、室温で3時間放置した。Ab(1-42)凝集体と結合した化合物と結合していない化合物とはWhatman GF/B filterを用いてBrandel M-24R cell harvesterによって分離した。濾過したフィルターに残った放射能をγ-カウンターで計測し、GraphPad Prism 4.0を用いて阻害曲線を作成し、阻害定数：K_iを算出した。 (6) Inhibition constant by binding inhibition experiment using Ab (1-42) aggregate: calculation of _Ki value
Mix 850 μL of 10% EtOH solution, 50 μL of 10% ethanol solution of [ ¹²⁵ I] IMPY, 50 μL (0-20 μM) of sample solution of various concentrations, and finally, Ab (1-42) aggregate solution 50 μL was added and left at room temperature for 3 hours. The compound bound to the Ab (1-42) aggregate and the compound not bound were separated by Brandel M-24R cell harvester using Whatman GF / B filter. The radioactivity remaining on the filtered filter was measured with a γ-counter, an inhibition curve was prepared using GraphPad Prism 4.0, and an inhibition constant: _Ki was calculated.

（７）正常マウスを用いた体内放射能分布実験
10%EtOH含有生理食塩水を用い、放射性ヨウ素標識体を希釈した。1群4-5匹の4-5週齢ddY雄系マウス (20-25 g) に、それぞれの標識体[¹²⁵I]7、[¹²⁵I]10、 [¹²⁵I]13、 [¹²⁵I]14を1匹あたり100 μL (0.2-0.4 μCi) 尾静脈より投与した。投与後2、10、30、60分後に断頭、採血し、主要な臓器を摘出した後、それらの重量および放射能を測定した。 (7) In vivo radioactivity distribution experiment using normal mice
The radioactive iodine label was diluted with physiological saline containing 10% EtOH. 4-5 week old ddY male mice (20-25 g) per group were each labeled [ ¹²⁵ I] 7, [ ¹²⁵ I] 10, [ ¹²⁵ I] 13, [ ¹²⁵ I] 14 was administered from the tail vein of 100 μL (0.2-0.4 μCi) per animal. 2, 10, 30, and 60 minutes after administration, the mice were decapitated and bled, and the main organs were removed, and their weight and radioactivity were measured.

（８）アルツハイマー病病態モデルマウス脳組織切片を用いた蛍光染色実験
アルツハイマー病モデルトランスジェニックマウスTg2576 (20,24ヶ月齢)の脳を取り出し、ドライアイス中で4％CMCを加え凍結させた。その後、LEICA CM1900により厚さ10 μmの切片を作成した。切片とリガンド（100μ M in 50％EtOH）を10分間、または切片とTh-Sを3分間反応させた後、50％EtOHで2回洗浄し、Nikon ECLIPSE 80iを用い、波長365 nm、380-420 nmで観察した。 (8) Fluorescence staining experiment using Alzheimer's disease state model mouse brain tissue section Alzheimer's disease model transgenic mouse Tg2576 (20, 24 months old) brain was taken out and frozen by adding 4% CMC in dry ice. Thereafter, a section having a thickness of 10 μm was prepared using LEICA CM1900. After reacting the section and ligand (100 μM in 50% EtOH) for 10 minutes or the section and Th-S for 3 minutes, wash with 50% EtOH twice and use Nikon ECLIPSE 80i at a wavelength of 365 nm, 380- Observed at 420 nm.

〔実験結果〕
（１）DPOD誘導体の合成
図１及び２にDPOD誘導体の合成経路を示す。DPOD骨格の形成は既報の方法を用いて行った。それぞれのブロモ化合物はパラジウムを触媒とするビス(トリブチルスズ)との反応によりトリブチルスズ体へと変換した。これらのトリブチルスズ体はヨウ素と反応させることによりヨウ素体へと変換した。〔Experimental result〕
(1) Synthesis of DPOD Derivative FIGS. 1 and 2 show the synthesis route of DPOD derivatives. The formation of the DPOD skeleton was carried out using the previously reported method. Each bromo compound was converted to a tributyltin compound by reaction with bis (tributyltin) using palladium as a catalyst. These tributyltin bodies were converted to iodine bodies by reacting with iodine.

（２） ¹²⁵ I標識実験
図３に2,5-ジフェニル-1,3,4-オキサジアゾール誘導体の¹²⁵I標識経路を示す。¹²⁵I標識は、過酸化水素を酸化剤として用い、スズ-ヨウ素交換反応により目的とする¹²⁵I標識体を得た。あらかじめ254 nmにおける非放射能化合物の吸光度を逆相HPLCで分析しておき、その保持時間と一致する化合物を分離精製することにより、目的とする¹²⁵I標識体を98%以上の放射化学的純度で得た。 (2) ¹²⁵ I labeling experiment FIG. 3 shows the ¹²⁵ I labeling pathway of 2,5-diphenyl-1,3,4-oxadiazole derivatives. ^{For 125} I labeling, hydrogen peroxide was used as an oxidizing agent, and the target ¹²⁵ I label was obtained by a tin-iodine exchange reaction. The absorbance of non-radioactive compounds in advance 254 nm leave analyzed by reverse phase by HPLC, the separation and purification of the compounds that match the retention time, ¹²⁵ I-labeled substance 98% or more radiochemical purity of interest Got in.

（３）DPOD誘導体のAb(1-42)凝集体との結合親和性に関する検討
合成した種々のDPOD誘導体（化合物12、13、14、15、16）について、阻害定数：K_iを算出するため、[¹²⁵I]IMPYを放射性リガンドとして用いた結合阻害実験を行った。実験の結果得られたK_iを表１７に示す。DPOD誘導体におけるAb(1-42)凝集体に対する結合親和性は、置換基の種類により異なり、OH <NH₂ = NHMe = NMe₂ < OMeの順で向上することが示された。これら化合物のKi値は4から47 nMであり、いずれの化合物も、インビボアミロイドイメージングを行うのに十分な結合性を示した。また、化合物19、20については、アミロイド凝集体との直接的な結合性を評価した（図４）。その結果、化合物19、20は、IMPYと同程度の結合性を有していることが明らかとなり、これら化合物もインビボアミロイドイメージングを行うのに十分な結合性を示した。 (3) DPOD various DPOD derivatives discussed synthesized for binding affinity to Ab (1-42) aggregates derivatives for (Compound 12,13,14,15,16), inhibition constants: To calculate a K _i , [ ¹²⁵ I] IMPY was used as a radioligand for binding inhibition experiments. Table 17 shows _Ki obtained as a result of the experiment. It was shown that the binding affinity for the Ab (1-42) aggregate in the DPOD derivative was different depending on the type of the substituent, and improved in the order of OH <NH ₂ = NHMe = NMe ₂ <OMe. The Ki values of these compounds were 4 to 47 nM, and all compounds showed sufficient binding properties for in vivo amyloid imaging. Moreover, about the compounds 19 and 20, the direct binding property with an amyloid aggregate was evaluated (FIG. 4). As a result, it was revealed that Compounds 19 and 20 have the same binding ability as IMPY, and these compounds also showed sufficient binding ability for in vivo amyloid imaging.

（４） ¹²⁵ I標識DPOD誘導体の正常マウスにおける体内放射能分布実験
4-5週齢正常マウスを用いて¹²⁵I標識化合物12、13、14、15、19、20のDPOD誘導体の体内放射能分布実験を行った（表１８）。いずれの化合物も1 %ID/g以上の脳移行性を示し、アミロイドイメージングに十分な放射能が脳へ到達した。また、1,2,4誘導体に比べ、1,3,4誘導体は高い脳移行性を示した。中でも[¹²⁵I]20は、最も速やかな放射能消失を示すことが確認された。いずれの化合物も肝臓への高い集積を示し、経時的に腸管へと排泄された。
(4) ^125I- labeled DPOD derivative in vivo distribution experiment in normal mice
In vivo radioactivity distribution experiments of DPOD derivatives of ¹²⁵ I-labeled compounds 12, 13, 14, 15, 19, and 20 were performed using 4-5 week old normal mice (Table 18). All of the compounds showed brain transferability of 1% ID / g or more, and radioactivity sufficient for amyloid imaging reached the brain. Compared with 1,2,4 derivatives, 1,3,4 derivatives showed higher brain migration. Among them, [ ¹²⁵ I] 20 was confirmed to show the most rapid loss of radioactivity. All compounds showed high accumulation in the liver and were excreted into the intestine over time.

（５）アルツハイマー病病態モデルマウス脳組織切片を用いた蛍光染色実験
図５に、アルツハイマー病マウス脳組織切片を用いた化合物19、20による蛍光染色の結果を示す。化合物19、20の蛍光染色部位は、アミロイド染色試薬であるチオフラビン-S の蛍光染色部位と一致しており、化合物19、20は、マウス脳アミロイド斑に対して結合親和性を持つことが示された。 (5) Fluorescence staining experiment using Alzheimer's disease state model mouse brain tissue section FIG. 5 shows the results of fluorescent staining with compounds 19 and 20 using Alzheimer's disease mouse brain tissue sections. The fluorescent staining sites of compounds 19 and 20 are consistent with the fluorescent staining sites of thioflavin-S, an amyloid staining reagent, indicating that compounds 19 and 20 have binding affinity for mouse brain amyloid plaques. It was.

2,5-ジフェニル-1,2,4-オキサジアゾール誘導体の合成経路を示す図。図中の番号は化合物の番号を示す。The figure which shows the synthetic pathway of a 2, 5- diphenyl -1,2, 4- oxadiazole derivative. The numbers in the figure indicate the compound numbers. 2,5-ジフェニル-1,3,4-オキサジアゾール誘導体の合成経路を示す図。図中の番号は化合物の番号を示す。The figure which shows the synthetic pathway of a 2, 5- diphenyl -1,3,4- oxadiazole derivative. The numbers in the figure indicate the compound numbers. 2,5-ジフェニル-1,3,4-オキサジアゾール誘導体の¹²⁵I標識経路を示す図。図中の番号は化合物の番号を示す。The figure which shows the ¹²⁵ I labeling route of a 2, 5- diphenyl -1,3,4- oxadiazole derivative. The numbers in the figure indicate the compound numbers. 化合物19および20のAβ42凝集体への結合性を示す図（化合物19：■、化合物20：●、IMPY：▲、コントロール：×）。The figure which shows the binding property to the A (beta) 42 aggregate of the compounds 19 and 20 (Compound 19: ■, Compound 20: ●, IMPY: ▲, control: X). アルツハイマー病モデルマウス脳切片を用いた化合物１９、２０による蛍光染色および隣接切片によるチオフラビンS染色の結果を示す図。The figure which shows the result of the fluorescent dyeing | staining by the compounds 19 and 20 which used the Alzheimer's disease model mouse brain section | slice, and the thioflavin S dyeing | staining by the adjacent section | slice.

Claims

Formula (I)
[Wherein, R ¹ represents a phenyl group which may be substituted with one or more substituents selected from the following substituent group A, and R ² represents 1 or 2 selected from the following substituent group B represents a more substituents which may be a phenyl group substituted by, substituent group a is a halogen atom, in the formula _{_{_{:-( CH 2 CH 2 O) n}}} -F [ wherein, n represents an integer of 1 to 10 To express. And a group represented by the formula: — (CH ₂ CH ₂ O) _n —OH [wherein n represents an integer of 1 to 10. The substituent group B is a halogen atom, a dimethylamino group, a methylamino group, an amino group, a methoxy group, a hydroxyl group, a formula: — (CH ₂ CH ₂ O) _n —F. [Wherein n represents an integer of 1 to 10. And a group represented by the formula: — (CH ₂ CH ₂ O) _n —OH [wherein n represents an integer of 1 to 10. ] Is a group consisting of groups represented by ]
Or general formula (II)
[Wherein, R ¹ and R ² are as defined above. ]
Or a compound obtained by labeling said compound with a radionuclide, or a pharmaceutically acceptable salt of these compounds.

The amyloid protein-related disease diagnostic composition according to claim 1, wherein the amyloid protein-related disease is an amyloid β protein-related disease.

The amyloid protein-related disease diagnostic composition according to claim 1, wherein the amyloid protein-related disease is Alzheimer's disease.

The composition for diagnosing amyloid protein-related diseases according to any one of claims 1 to 3, wherein R ¹ in the general formulas (I) and (II) is a phenyl group substituted with a fluorine atom or an iodine atom.

The composition for diagnosing amyloid protein-related diseases according to any one of claims 1 to 3, wherein R ¹ in the general formulas (I) and (II) is a 4-fluorophenyl group or a 4-iodophenyl group. .

The R ² in the general formulas (I) and (II) is a dimethylamino group, a methylamino group, an amino group, a methoxy group, or a phenyl group substituted with a hydroxyl group. A composition for diagnosing amyloid protein-related diseases.

The amyloid according to any one of claims 1 to 5, wherein R ² in the general formulas (I) and (II) is a phenyl group substituted with a dimethylamino group, a methylamino group, an amino group, or a methoxy group. A composition for diagnosis of sex protein-related diseases.

The composition for diagnosing amyloid protein-related diseases according to any one of claims 1 to 5, wherein R ² in the general formulas (I) and (II) is a phenyl group substituted with a methoxy group.

The composition for diagnosing amyloid protein-related diseases according to any one of claims 1 to 5, wherein R ² in the general formulas (I) and (II) is a 4-methoxyphenyl group.

The composition for diagnosing amyloid protein-related diseases according to any one of claims 1 to 9, wherein the radionuclide is a positron emitting nuclide.

The composition for diagnosing amyloid protein-related diseases according to any one of claims 1 to 9, wherein the radionuclide is a γ-ray emitting nuclide.

A step of administering a test substance to a model animal of amyloid protein-related disease, a step of administering the amyloid protein-related disease diagnostic composition according to any one of claims 1 to 11 to the model animal, and the model A screening method for a therapeutic or prophylactic agent for amyloid protein-related diseases, comprising a step of examining the distribution or amount of a compound represented by the general formula (I) or (II) contained in an animal brain.

A step of administering a therapeutic or prophylactic agent for the disease to a model animal of amyloid protein-related disease, and administering the composition for diagnosing amyloid protein-related disease according to any one of claims 1 to 11 to the model animal. And a method for evaluating a therapeutic or prophylactic agent for amyloid protein-related diseases, comprising the step of examining the distribution or amount of the compound represented by the general formula (I) or (II) contained in the brain of the model animal .