JP5424296B2

JP5424296B2 - Method for producing flavone derivative and sialyltransferase inhibitor

Info

Publication number: JP5424296B2
Application number: JP2008219288A
Authority: JP
Inventors: 敏幸菅; 隆鈴木; 一八左; 巧古田
Original assignee: Shizuoka University NUC
Current assignee: Shizuoka University NUC
Priority date: 2008-08-28
Filing date: 2008-08-28
Publication date: 2014-02-26
Anticipated expiration: 2028-08-28
Also published as: JP2010053067A

Description

本発明は、フラボン誘導体の製造方法、ならびにフラボン誘導体を有効成分とするシアル酸転移酵素阻害剤に関する。 The present invention relates to a method for producing a flavone derivative and a sialyltransferase inhibitor containing the flavone derivative as an active ingredient.

シアル酸転移酵素は生体内で糖蛋白質や糖脂質の糖鎖にシアル酸を転移する酵素であり、癌の進行や転移、炎症、免疫応答、ウイルス感染などにおける関与が示唆されている。しかしながら、シアル酸転移酵素に対して阻害活性を有する物質はほとんど知られていない。特開平８−１９８８９１および特開平８−１９８８９２は、シアル酸転移酵素阻害活性をもつシアル酸誘導体を開示する。また、特開平１０−３３０３６４は、ペニシリウム属の微生物により産生されるある種のリン脂質がシアル酸転移酵素阻害活性をもつことを開示する。シアル酸転移酵素阻害活性を有するフラボン誘導体やカテキン誘導体はこれまでに全く報告がない。
特開平８−１９８８９１特開平８−１９８８９２特開平１０−３３０３６４ Sialyltransferase is an enzyme that transfers sialic acid to sugar chains of glycoproteins and glycolipids in vivo, and has been suggested to be involved in cancer progression, metastasis, inflammation, immune response, and viral infection. However, few substances have inhibitory activity against sialyltransferases. JP-A-8-198891 and JP-A-8-198892 disclose sialic acid derivatives having sialyltransferase inhibitory activity. JP-A-10-330364 discloses that certain phospholipids produced by Penicillium microorganisms have sialyltransferase inhibitory activity. There have been no reports of flavone derivatives or catechin derivatives having sialic acid transferase inhibitory activity so far.
JP-A-8-198891 JP-A-8-198892 JP 10-330364 A

本発明者らは、茶の成分であるカテキン類やテアフラビン類の種々の誘導体を化学合成してシアル酸転移酵素阻害活性を測定したところ、驚くべきことに、ある種のフラボン誘導体が高いシアル酸転移酵素阻害活性を有することを見いだした。さらにこれらのフラボン誘導体を合成する方法を見いだして、本発明を完成させた。 The inventors of the present invention chemically synthesized various derivatives of catechins and theaflavins, which are components of tea, and measured sialyltransferase inhibitory activity. Surprisingly, some flavone derivatives had high sialic acid derivatives. It was found to have transferase inhibitory activity. Furthermore, the inventors have found a method for synthesizing these flavone derivatives and completed the present invention.

すなわち、本発明は、一般式（I）：
［式中、Ｒ_１はＨまたはＯＨであり、ｎは１、２または３である］
で表される化合物を製造する方法を提供する。この方法は、
（ａ）次式（II）：
で表される化合物の水酸基を保護して、次式（III）：
［式中、Ｒ_２は水酸基の保護基である］
で表される化合物を製造し；
（ｂ）次式（IV）：
［式中、ｎは上で定義されるとおりである］
で表される化合物に保護基を導入して、次式（V）：
［式中、Ｒ_３は水酸基の保護基であり、Ｒ_４はアシル基の保護基であり、ｎは上で定義されるとおりである］
で表される化合物を製造し；
（ｃ）式（III）の化合物と式（V）の化合物を反応させて、次式（VI）：
［式中、Ｒ_２、Ｒ_３およびｎは上で定義されるとおりである］
で表される化合物を製造し；
（ｄ）式（VI）の化合物の保護基Ｒ_２を除去して、次式（VII）：
［式中、Ｒ_３およびｎは上で定義されるとおりである］
で表される化合物を製造し；
（ｅ）式（VII）の化合物を環化させて、次式（VIII）：
［式中、Ｒ_３およびｎは上で定義されるとおりである］
で表される化合物を製造し；
（ｆ）任意に、式（VIII）の化合物のクロモン環の３位を酸化して、次式（VIII’）：
［式中、Ｒ_３およびｎは上で定義されるとおりである］
で表される化合物を製造し；
（ｇ）式（VIII）または（VIII’）の化合物の水酸基の保護基を除去して、一般式（I）：
の化合物を得る、
の各工程を含む方法を提供する。 That is, the present invention relates to the general formula (I):
[Wherein R ₁ is H or OH, and n is 1, 2 or 3]
The method of manufacturing the compound represented by this is provided. This method
(A) The following formula (II):
Protecting the hydroxyl group of the compound represented by the following formula (III):
[Wherein R ₂ is a hydroxyl-protecting group]
A compound represented by:
(B) The following formula (IV):
[Wherein n is as defined above]
A protective group is introduced into the compound represented by the following formula (V):
[Wherein R ₃ is a protecting group for a hydroxyl group, R ₄ is a protecting group for an acyl group, and n is as defined above]
A compound represented by:
(C) reacting a compound of formula (III) with a compound of formula (V) to give the following formula (VI):
[Wherein R ₂ , R ₃ and n are as defined above]
A compound represented by:
(D) removing the protecting group R ₂ of the compound of formula (VI) to give the following formula (VII):
[Wherein R ₃ and n are as defined above]
A compound represented by:
(E) The compound of formula (VII) is cyclized to give the following formula (VIII):
[Wherein R ₃ and n are as defined above]
A compound represented by:
(F) optionally oxidizing the 3-position of the chromone ring of the compound of formula (VIII) to give the following formula (VIII ′):
[Wherein R ₃ and n are as defined above]
A compound represented by:
(G) removing the protecting group for the hydroxyl group of the compound of formula (VIII) or (VIII ′) to give a compound of the general formula (I):
To obtain a compound of
The method including each of these steps is provided.

好ましくは、本発明の方法において製造される一般式（Ｉ）の化合物は、下記のいずれかの化合物：
である。 Preferably, the compound of general formula (I) produced in the process of the present invention is one of the following compounds:
It is.

別の観点においては、本発明は、下記のいずれかの式：
で表される化合物を有効成分とするシアル酸転移酵素阻害剤を提供する。 In another aspect, the invention provides any of the following formulas:
The sialyltransferase inhibitor which uses the compound represented by these as an active ingredient is provided.

本発明によりシアル酸転移酵素阻害活性を有するフラボン誘導体を簡便かつ大量に合成することが可能となる。本発明のシアル酸転移酵素阻害剤は、炎症、がん転移、ウイルス感染症の予防／治療剤として、例えばインフルエンザの予防/治療剤として、あるいはこれらの疾患の発症メカニズムの解明に有用である。 According to the present invention, a flavone derivative having sialyltransferase inhibitory activity can be synthesized easily and in large quantities. The sialyltransferase inhibitor of the present invention is useful as a prophylactic / therapeutic agent for inflammation, cancer metastasis, and viral infection, for example, as a prophylactic / therapeutic agent for influenza, or for elucidating the onset mechanism of these diseases.

本発明は、一般式（I）：
［式中、Ｒ_１はＨまたはＯＨであり、ｎは１、２または３である］
で表される化合物を製造する方法を提供する。式（I）のフラボン誘導体は、天然のフラボンまたはフラボノールと異なり、Ａ環に水酸基を有さないことを特徴とする。驚くべきことに、このような構造を有するフラボン誘導体が、高いシアル酸転移酵素阻害活性を有することが見いだされた。 The present invention is directed to general formula (I):
[Wherein R ₁ is H or OH, and n is 1, 2 or 3]
The method of manufacturing the compound represented by this is provided. Unlike natural flavones or flavonols, the flavone derivative of formula (I) is characterized by having no hydroxyl group in the A ring. Surprisingly, it has been found that flavone derivatives having such a structure have high sialyltransferase inhibitory activity.

本発明の製造方法においては、出発物質として、式：
で表される２’−ヒドロキシアセトフェノンを用いる。この化合物は市販されている。 In the production method of the present invention, the starting material is represented by the formula:
2′-hydroxyacetophenone represented by This compound is commercially available.

本発明の方法の工程（ａ）においては、式（II）で表される化合物の水酸基を保護して、次式（III）：
［式中、Ｒ_２は水酸基の保護基である］
で表される化合物を製造する。 In step (a) of the method of the present invention, the hydroxyl group of the compound represented by formula (II) is protected, and the following formula (III):
[Wherein R ₂ is a hydroxyl-protecting group]
The compound represented by these is manufactured.

Ｒ_２としては、フェノール性水酸基の保護基として一般に使用されるいずれの基を用いてもよく、例えば、メタンスルホニル基、トリフルオロメタンスルホニル基、ベンゼンスルホニル基、トルエンスルホニル基、ニトロベンゼンスルホニル基、アセチル基、トリクロロアセチル基、トリフルオロアセチル基、ベンゾイル基、ピバロイル基、直鎖または分枝鎖のＣ_１−４アルキル基、ベンジル基、トリメチルシリル基、トリエチルシリル基、t-ブチルジメチルシリル基、t-ブチルジフェニルシリル基などが挙げられる。Ｒ_２として特に好ましいものはt-ブチルジフェニルシリル基である。 As R ₂ , any group generally used as a protecting group for a phenolic hydroxyl group may be used. For example, methanesulfonyl group, trifluoromethanesulfonyl group, benzenesulfonyl group, toluenesulfonyl group, nitrobenzenesulfonyl group, acetyl group , Trichloroacetyl group, trifluoroacetyl group, benzoyl group, pivaloyl group, linear or branched C _1-4 alkyl group, benzyl group, trimethylsilyl group, triethylsilyl group, t-butyldimethylsilyl group, t-butyl Examples thereof include a diphenylsilyl group. Particularly preferred as R ₂ is a t-butyldiphenylsilyl group.

反応は、非プロトン性溶媒中で塩基の存在下で、式（II）の化合物を式：Ｒ_２−Ｌ（Ｌは脱離基である）と反応させることにより行う。非プロトン性溶媒としては、アセトニトリル、塩化メチレン、トルエン、THF、DMF等が挙げられる。塩基としては、トリエチルアミン、ジイソプロピルエチルアミン、イミダゾール等の有機塩基や炭酸カリウム等の無機塩基を用いることができる。 The reaction is carried out by reacting the compound of formula (II) with the formula: R ₂ -L (L is a leaving group) in the presence of a base in an aprotic solvent. Examples of the aprotic solvent include acetonitrile, methylene chloride, toluene, THF, DMF and the like. As the base, organic bases such as triethylamine, diisopropylethylamine and imidazole, and inorganic bases such as potassium carbonate can be used.

本発明の方法においては、別の出発物質として次式（IV）：
［式中、ｎは上で定義されるとおりである］
で表される化合物を用いる。式（IV）の化合物としては、例えば、４−ヒドロキシ安息香酸、３，４−ジヒドロキシ安息香酸、３，４−ジヒドロキシ安息香酸、没食子酸などが挙げられ、いずれも市販されている。 In the process of the present invention, another starting material is the following formula (IV):
[Wherein n is as defined above]
The compound represented by these is used. Examples of the compound of formula (IV) include 4-hydroxybenzoic acid, 3,4-dihydroxybenzoic acid, 3,4-dihydroxybenzoic acid, gallic acid and the like, and all are commercially available.

本発明の方法の工程（ｂ）においては、式（IV）で表される化合物に保護基を導入して、次式（V）：
［式中、Ｒ_３は水酸基の保護基であり、Ｒ_４はアシル基の保護基であり、ｎは上で定義されるとおりである］
で表される化合物を製造する。 In step (b) of the method of the present invention, a protecting group is introduced into the compound represented by formula (IV), and the following formula (V):
[Wherein R ₃ is a protecting group for a hydroxyl group, R ₄ is a protecting group for an acyl group, and n is as defined above]
The compound represented by these is manufactured.

Ｒ_３としては、フェノール性水酸基の保護基として一般に使用されるいずれの基、例えば、上述のＲ_２として用いられる任意の基を用いることができる。なお、Ｒ_２とＲ_３は、後の工程（ｄ）において保護基Ｒ_２を脱保護する条件下でＲ_３が脱保護されないように選択する。 As R ₃ , any group generally used as a protecting group for a phenolic hydroxyl group, for example, any group used as R ₂ described above can be used. R ₂ and R ₃ are selected so that R ₃ is not deprotected under the conditions for deprotecting the protecting group R ₂ in the subsequent step (d).

Ｒ_４としては、アシル基の保護基として一般に使用される任意の基を用いることができる。Ｒ_４として特に好ましいものはベンゾトリアゾールである。ベンゾトリアゾールを用いる場合、反応は、式（V）の化合物をＲ_３基で保護した後、非プロトン性溶媒中で塩化チオニルと反応させて酸クロリドとし、非プロトン性溶媒中で塩基の存在下でベンゾトリアゾールと反応させることにより行う。 As R ₄ , any group generally used as a protecting group for an acyl group can be used. Particularly preferred as R ₄ is benzotriazole. When benzotriazole is used, the reaction is carried out by protecting the compound of formula (V) with the R ₃ group and then reacting with thionyl chloride in an aprotic solvent to give an acid chloride in the presence of a base in the aprotic solvent. By reacting with benzotriazole.

本発明の方法の工程（ｃ）においては、それぞれ工程（ａ）および（ｂ）で得られた式（III）の化合物と式（V）の化合物を反応させて、次式（VI）：
［式中、Ｒ_２、Ｒ_３およびｎは上で定義されるとおりである］
で表される化合物を製造する。反応は、非プロトン性溶媒中で塩基の存在下で行い、好ましくは低温下で行う。塩基としては、例えば、リチウムヘキサメチルジシラジドを用いることができる。 In step (c) of the method of the present invention, the compound of formula (III) obtained in steps (a) and (b) is reacted with the compound of formula (V) to give the following formula (VI):
[Wherein R ₂ , R ₃ and n are as defined above]
The compound represented by these is manufactured. The reaction is performed in the presence of a base in an aprotic solvent, preferably at a low temperature. As the base, for example, lithium hexamethyldisilazide can be used.

本発明の方法の工程（ｄ）においては、式（VI）の化合物の保護基Ｒ_２を除去して、次式（VII）：
［式中、Ｒ_３およびｎは上で定義されるとおりである］
で表される化合物を製造する。 In step (d) of the process of the invention, the protecting group R ₂ of the compound of formula (VI) is removed to give the following formula (VII):
[Wherein R ₃ and n are as defined above]
The compound represented by these is manufactured.

反応は、通常の脱保護条件下であって、Ｒ_２のみが脱保護され、Ｒ_３は脱保護されないような条件下で行う。当業者は、Ｒ_２とＲ_３の組み合わせに基づいて、このような条件を容易に選定することができる。なお、この工程は酢酸の存在下で行うことにより、副生成物の生成を抑制することができる。 The reaction is carried out under normal deprotection conditions, such that only R ₂ is deprotected and R ₃ is not deprotected. Those skilled in the art can easily select such conditions based on the combination of R ₂ and R ₃ . In addition, the production | generation of a by-product can be suppressed by performing this process in presence of acetic acid.

次に、本発明の方法の工程（ｅ）においては、式（VII）の化合物を環化させて、次式（VIII）：
［式中、Ｒ_３およびｎは上で定義されるとおりである］
で表される化合物を製造する。環化反応は、非プロトン性溶媒中で酸性条件下で、トリフルオロ酢酸等の酸化剤を用いて行う。この環化反応により、Ａ環に水酸基を有さないフラボノイド骨格を得ることができる。 Next, in step (e) of the method of the present invention, the compound of formula (VII) is cyclized to give the following formula (VIII):
[Wherein R ₃ and n are as defined above]
The compound represented by these is manufactured. The cyclization reaction is performed using an oxidizing agent such as trifluoroacetic acid under acidic conditions in an aprotic solvent. By this cyclization reaction, a flavonoid skeleton having no hydroxyl group in the A ring can be obtained.

本発明の方法の工程（ｆ）においては、式（VIII）の化合物のクロモン環の３位を酸化して、次式（VIII’）：
［式中、Ｒ_３およびｎは上で定義されるとおりである］
で表される化合物を製造する。工程（ｆ）は任意の工程であり、式（I）においてＲ_１がＯＨである場合に実施する。反応は、非プロトン性溶媒中でジメチルジオキシラン等の酸化剤を用いて行う。 In step (f) of the method of the present invention, the 3-position of the chromone ring of the compound of formula (VIII) is oxidized to give the following formula (VIII ′):
[Wherein R ₃ and n are as defined above]
The compound represented by these is manufactured. Step (f) is an optional step and is carried out when R ₁ is OH in formula (I). The reaction is carried out in an aprotic solvent using an oxidizing agent such as dimethyldioxirane.

本発明の方法の工程（ｇ）においては、工程（ｅ）または（ｆ）で得られた式（VIII）または（VIII’）の化合物の水酸基の保護基を除去して、目的とする一般式（I）：
の化合物を得る。反応は、通常の脱保護条件下で行う。 In step (g) of the method of the present invention, the protective group for the hydroxyl group of the compound of formula (VIII) or (VIII ′) obtained in step (e) or (f) is removed to obtain the desired general formula. (I):
To obtain a compound of The reaction is carried out under normal deprotection conditions.

以上のようにして得られる一般式（I）の化合物としては、例えば下記の化合物：
が挙げられる。 Examples of the compound of the general formula (I) obtained as described above include the following compounds:
Is mentioned.

本発明においては、上記の化合物がいずれも高いシアル酸転移酵素阻害活性を有することが見いだされた。すなわち、別の観点において本発明は、以下の化合物：
を有効成分とするシアル酸転移酵素阻害剤を提供する。 In the present invention, it has been found that any of the above compounds has high sialyltransferase inhibitory activity. That is, in another aspect, the present invention provides the following compounds:
The present invention provides a sialyltransferase inhibitor containing as an active ingredient.

本発明にしたがって得られる化合物のシアル酸転移酵素阻害活性は、定法にしたがって測定することができる。例えば、下記の実施例に示されるように、プレート表面に糖鎖含有ポリマーをコーティングしておき、ここに種々の濃度の試験化合物の存在下でシアル酸転移酵素と基質であるCMP-Neu5Acとを加えてインキュベーションする。プレートを洗浄した後、標識したアグルチニンを加えて、糖鎖に付加したシアル酸を定量することにより、試験化合物のシアル酸転移酵素阻害活性を測定することができる。 The sialyltransferase inhibitory activity of the compound obtained according to the present invention can be measured according to a conventional method. For example, as shown in the following examples, a sugar chain-containing polymer is coated on the plate surface, and sialyltransferase and the substrate CMP-Neu5Ac are added thereto in the presence of various concentrations of the test compound. In addition, incubate. After washing the plate, labeled agglutinin is added, and the sialic acid added to the sugar chain is quantified, whereby the sialic acid transferase inhibitory activity of the test compound can be measured.

本発明において、シアル酸転移酵素阻害剤とは、本発明にしたがうフラボン誘導体または薬学的に許容しうる塩を、薬学的に許容しうる担体もしくは賦形剤とともに含む医薬組成物を表す。シアル酸転移酵素は生体内で糖蛋白質や糖脂質の糖鎖にシアル酸を転移することから、本発明のシアル酸転移酵素阻害剤は、炎症、がん転移、ウイルス感染症の予防／治療や、その発症メカニズムの解明に有用である。 In the present invention, a sialyltransferase inhibitor represents a pharmaceutical composition comprising a flavone derivative or a pharmaceutically acceptable salt according to the present invention together with a pharmaceutically acceptable carrier or excipient. Since sialyltransferases transfer sialic acid to glycoproteins or glycolipid sugar chains in vivo, the sialyltransferase inhibitors of the present invention are useful for the prevention / treatment of inflammation, cancer metastasis, and viral infections. It is useful for elucidating the onset mechanism.

薬学的に許容しうる塩としては、薬理学的に許容されるものであれば特に限定されず、例えば、ナトリウム、カリウム、カルシウム等のアルカリ金属またはアルカリ土類金属等の塩、アンモニアや各種有機塩基等の塩類を挙げることができる。 The pharmaceutically acceptable salt is not particularly limited as long as it is pharmacologically acceptable, and examples thereof include salts of alkali metals or alkaline earth metals such as sodium, potassium and calcium, ammonia and various organic substances. Examples thereof include salts such as bases.

本発明のシアル酸転移酵素阻害剤は、当業者に公知の方法で製剤化することができる。例えば、薬学的に許容しうる担体もしくは賦形剤、具体的には、滅菌水や生理食塩水、植物油、乳化剤、懸濁剤、界面活性剤、安定剤、香味剤、ベヒクル、防腐剤、結合剤などと適宜組み合わせて、一般に認められた製薬実施に要求される単位用量形態で混和することによって製剤化することができる。 The sialyltransferase inhibitor of the present invention can be formulated by methods known to those skilled in the art. For example, a pharmaceutically acceptable carrier or excipient, specifically, sterile water or saline, vegetable oil, emulsifier, suspending agent, surfactant, stabilizer, flavoring agent, vehicle, preservative, binding It can be formulated by mixing in appropriate unit dosage forms required for generally accepted pharmaceutical practice, in appropriate combination with agents.

本発明のシアル酸転移酵素阻害剤の適当な投与経路には、限定されないが、経口、直腸内、経粘膜、または腸内投与、または筋肉内、皮下、骨髄内、鞘内、直接心室内、静脈内、硝子体内、腹腔内、鼻腔内、または眼内注射が含まれる。投与経路および投与方法は、患者の年齢、症状により適宜選択することができる。経口または経鼻投与が好ましい。経口投与用には，例えば，化合物をカプセル剤，錠剤および液体製剤（シロップ剤，エリキシル剤および濃縮ドロップ剤など）のような慣用の経口投与形に製剤することができる。吸入用には，本発明の化合物を乾燥粉体または適当な溶液，懸濁液，またはエアロゾルとして製剤することができる。粉体および溶液は，当該技術分野において知られる適当な添加物とともに製剤することができる。非経口投与用には、本発明の化合物またはその塩を当該技術分野においてよく知られる薬学的に許容しうるベヒクルを用いて通常の製剤実施に従って処方することができる。 Suitable routes of administration of the sialyltransferase inhibitors of the present invention include, but are not limited to, oral, rectal, transmucosal, or enteral administration, or intramuscular, subcutaneous, intramedullary, intrathecal, direct ventricular, Intravenous, intravitreal, intraperitoneal, intranasal, or intraocular injection is included. The administration route and administration method can be appropriately selected depending on the age and symptoms of the patient. Oral or nasal administration is preferred. For oral administration, for example, the compounds can be formulated into conventional oral dosage forms such as capsules, tablets, and liquid preparations such as syrups, elixirs and concentrated drops. For inhalation, the compounds of the invention can be formulated as a dry powder or a suitable solution, suspension, or aerosol. Powders and solutions can be formulated with suitable additives known in the art. For parenteral administration, the compounds of the invention or salts thereof can be formulated according to conventional pharmaceutical practice using pharmaceutically acceptable vehicles well known in the art.

本発明のシアル酸転移酵素阻害剤の投与量としては、例えば、一回につき体重1kgあたり0.0001mgから1000mgの範囲で選ぶことが可能である。あるいは、例えば、患者あたり0.001〜100000mg/bodyの範囲で投与量を選ぶことができるが、これらの数値に必ずしも制限されるものではない。投与量、投与方法は、患者の体重や年齢、症状などにより変動するが、当業者であれば適宜選択することが可能である。 The dosage of the sialyltransferase inhibitor of the present invention can be selected, for example, in the range of 0.0001 mg to 1000 mg per kg of body weight at a time. Alternatively, for example, the dose can be selected in the range of 0.001 to 100000 mg / body per patient, but is not necessarily limited to these values. The dose and administration method vary depending on the weight, age, symptoms, etc. of the patient, but can be appropriately selected by those skilled in the art.

以下に実施例を挙げ、本発明をさらに詳しく説明する。ただし、本発明はこれに限定されるものではない。 The following examples further illustrate the present invention. However, the present invention is not limited to this.

実施例１フラボン誘導体の製造方法
1-[2-(tert-ブチルジフェニルシリルオキシ)フェニル]エタノン(2)の合成
Ar 雰囲気下、0 °C で 1 (5.0 g, 36.7 mmol) に DMF (7.3 mL)、塩化tert-ブチルジフェニルシリル(11.1 g, 40.4 mmol)、イミダゾール(3.75 g, 55.1 mmol) を加え、100 °Cで 4 時間攪拌した。飽和塩化アンモニウム水溶液を加えた後、n-ヘキサンで三回抽出し、有機層を飽和食塩水で洗浄、MgSO₄で乾燥、減圧下濃縮した。その後、再結晶 (n-ヘキサン / EtOAc) により精製し、無色個体の 1-[2-(tert-ブチルジフェニルシリルオキシ)フェニル]エタノン(2) (7.65 g, 56%) を得た。
¹H NMR (500 MHz, CDCl₃): δ 7.73 (d, J = 7.3 Hz, 1H), 7.58 (dd, J = 7.3, 1.8 Hz, 1H), 7.50-7.30 (m, 6H), 6.96 (td, J= 7.3, 1.8 Hz, 1H), 6.88 (t, J= 7.3 Hz, 1H), 6.47 (d, J = 7.3 Hz, 1H), 2.77 (s, 3H), 1.09 (s, 9H) Example 1 Method for producing flavone derivative
Synthesis of 1- [2- (tert-butyldiphenylsilyloxy) phenyl] ethanone (2)
DMF (7.3 mL), tert-butyldiphenylsilyl chloride (11.1 g, 40.4 mmol), imidazole (3.75 g, 55.1 mmol) were added to 1 (5.0 g, 36.7 mmol) at 0 ° C under Ar atmosphere, and 100 ° C Stir at C for 4 hours. A saturated aqueous ammonium chloride solution was added, and the mixture was extracted 3 times with n-hexane. The organic layer was washed with saturated brine, dried over MgSO ₄ , and concentrated under reduced pressure. Then, purification by recrystallization (n-hexane / EtOAc) gave colorless solid 1- [2- (tert-butyldiphenylsilyloxy) phenyl] ethanone (2) (7.65 g, 56%).
¹ H NMR (500 MHz, CDCl ₃ ): δ 7.73 (d, J = 7.3 Hz, 1H), 7.58 (dd, J = 7.3, 1.8 Hz, 1H), 7.50-7.30 (m, 6H), 6.96 (td , J = 7.3, 1.8 Hz, 1H), 6.88 (t, J = 7.3 Hz, 1H), 6.47 (d, J = 7.3 Hz, 1H), 2.77 (s, 3H), 1.09 (s, 9H)

(1H-ベンゾ[d][1,2,3]トリアゾール-1-イル)[4-(ベンジルオキシ)フェニル]メタノン (5) の合成
Ar 雰囲気下、0 °C で 3 (10.0 g, 43.8 mmol) に CH₂Cl₂(100 mL)、SOCl₂(4.8 mL, 65.7 mmol)、DMF (1.4 mL, 17.5 mmol) を加え、0 °C で 2 時間攪拌した。その後、減圧下濃縮し、無色個体の塩化4-(ベンジルオキシ)ベンゾイル(4) を含む粗精製物を得た。続いて、Ar 雰囲気下、0 °C で 4 を含む粗精製物にCH₂Cl₂ (150 mL)、Et₃N (6.1 mL, 44 mmol)、1H-ベンゾ[d][1,2,3]トリアゾール(5.2 g, 43.8 mmol) を加え、0 °C で 1 時間攪拌した。飽和塩化アンモニウム水溶液を加えた後、EtOAc で三回抽出し、有機層を飽和食塩水で洗浄、MgSO₄で乾燥、減圧下濃縮した。その後、再結晶 (n-ヘキサン / CH₂Cl₂) により精製し、無色個体の (1H-ベンゾ[d][1,2,3]トリアゾール-1-イル)[4-(ベンジルオキシ)フェニル]メタノン (5) (9.1 g, 2 工程 62%) を得た。
¹H NMR (500 MHz, CDCl₃): δ 8.38 (d, J= 8.0 Hz, 1H), 8.30 (d, J = 9.2 Hz, 2H), 8.16 (d, J = 8.0 Hz, 1H), 7.69 (t, J = 8.0 Hz, 1H), 7.54 (t, J = 8.0 Hz, 1H), 7.50-7.30 (m, 5H), 7.14 (d, J = 9.2 Hz, 2H), 5.21 (s, 2H) Synthesis of (1H-benzo [d] [1,2,3] triazol-1-yl) [4- (benzyloxy) phenyl] methanone (5)
Under Ar atmosphere, add CH ₂ Cl ₂ (100 mL), SOCl ₂ (4.8 mL, 65.7 mmol), DMF (1.4 mL, 17.5 mmol) to 3 (10.0 g, 43.8 mmol) at 0 ° C, then add 0 ° C For 2 hours. Thereafter, the mixture was concentrated under reduced pressure to obtain a crude product containing colorless solid 4- (benzyloxy) benzoyl chloride (4). Subsequently, CH ₂ Cl ₂ (150 mL), Et ₃ N (6.1 mL, 44 mmol), 1H-benzo [d] [1,2,3 were added to the crude product containing 4 at 0 ° C under Ar atmosphere. ] Triazole (5.2 g, 43.8 mmol) was added, and the mixture was stirred at 0 ° C for 1 hour. Saturated aqueous ammonium chloride solution was added, and the mixture was extracted 3 times with EtOAc. The organic layer was washed with saturated brine, dried over MgSO ₄ , and concentrated under reduced pressure. Then, it was purified by recrystallization (n-hexane / CH ₂ Cl ₂ ), and colorless solid (1H-benzo [d] [1,2,3] triazol-1-yl) [4- (benzyloxy) phenyl] Methanone (5) (9.1 g, 2 steps 62%) was obtained.
¹ H NMR (500 MHz, CDCl ₃ ): δ 8.38 (d, J = 8.0 Hz, 1H), 8.30 (d, J = 9.2 Hz, 2H), 8.16 (d, J = 8.0 Hz, 1H), 7.69 ( t, J = 8.0 Hz, 1H), 7.54 (t, J = 8.0 Hz, 1H), 7.50-7.30 (m, 5H), 7.14 (d, J = 9.2 Hz, 2H), 5.21 (s, 2H)

1-[4-(ベンジルオキシ)フェニル]-3-[2-(tert-ブチルジフェニルシリルオキシ)フェニル]プロパン-1,3-ジオン (6) の合成
Ar 雰囲気下、-78 °C で 2 (1.0 g, 2.7 mmol) に THF (14 mL)、5 (879 mg, 2.67 mmol)、LiHMDS (14 mL, 14 mmol, 1 M溶液、THF中) を加え、-78 °C で 1 時間攪拌した。水を加えた後、EtOAc で三回抽出し、有機層を飽和食塩水で洗浄、MgSO₄ で乾燥、減圧下濃縮した。その後、シリカゲルカラムクロマトグラフィー (n-ヘキサン : EtOAc = 20 : 1) により精製し、白色固体の 1-[4-(ベンジルオキシ)フェニル]-3-[2-(tert-ブチルジフェニルシリルオキシ)フェニル]プロパン-1,3-ジオン (6) (1.56 g, quant.) を得た。
¹H NMR (500 MHz, CDCl₃): δ 17.0 (s, 1H), 7.88 (d, J = 9.2 Hz, 2H), 7.75-7.70 (m, 5H), 7.50-7.30 (m, 11H), 6.56 (dd, J = 8.0, 1.2 Hz, 1H), 5.13 (s, 2H), 1.03 (s, 9H) Synthesis of 1- [4- (benzyloxy) phenyl] -3- [2- (tert-butyldiphenylsilyloxy) phenyl] propane-1,3-dione (6)
Add THF (14 mL), 5 (879 mg, 2.67 mmol), LiHMDS (14 mL, 14 mmol, 1 M solution in THF) to 2 (1.0 g, 2.7 mmol) at -78 ° C under Ar atmosphere. The mixture was stirred at -78 ° C for 1 hour. Water was added, and the mixture was extracted 3 times with EtOAc. The organic layer was washed with saturated brine, dried over MgSO ₄ , and concentrated under reduced pressure. After that, it was purified by silica gel column chromatography (n-hexane: EtOAc = 20: 1), and 1- [4- (benzyloxy) phenyl] -3- [2- (tert-butyldiphenylsilyloxy) phenyl as a white solid Propane-1,3-dione (6) (1.56 g, quant.) Was obtained.
¹ H NMR (500 MHz, CDCl ₃ ): δ 17.0 (s, 1H), 7.88 (d, J = 9.2 Hz, 2H), 7.75-7.70 (m, 5H), 7.50-7.30 (m, 11H), 6.56 (dd, J = 8.0, 1.2 Hz, 1H), 5.13 (s, 2H), 1.03 (s, 9H)

1-[4-(ベンジルオキシ)フェニル]-3-(2-ヒドロキシフェニル)プロパン-1,3-ジオン (7)の合成
室温で 6 (450 mg, 0.77 mmol) に THF (2.5 mL)、AcOH (44μL, 0.77 mmol)、TBAF (0.8 mL, 0.8 mmol, 1M溶液、THF中) を加え、室温で 5 分間攪拌した。飽和塩化アンモニウム水溶液を加えた後、Et₂O で三回抽出し、有機層を飽和食塩水で洗浄、MgSO₄で乾燥、減圧下濃縮した。その後、シリカゲルカラムクロマトグラフィー (n-ヘキサン : EtOAc = 10 : 1) により精製し、黄色個体の1-[4-(ベンジルオキシ)フェニル]-3-(2-ヒドロキシフェニル)プロパン-1,3-ジオン (7) (260 mg, 98%) を得た。
¹H NMR (500 MHz, CDCl₃): δ 15.8 (s, 1H), 12.1 (s, 1H), 7.92 (d, J = 9.2 Hz, 2H), 7.77 (dd, J = 8.6, 1.8 Hz, 1H), 7.50-7.30 (m, 6H), 7.06 (d, J = 9.2 Hz, 2H), 7.00 (dd, J = 8.6, 1.8 Hz, 1H), 6.91 (t, J = 8.6 Hz, 1H), 6.77 (s, 1H), 5.16 (s, 2H) Synthesis of 1- [4- (benzyloxy) phenyl] -3- (2-hydroxyphenyl) propane-1,3-dione (7)
THF (2.5 mL), AcOH (44 μL, 0.77 mmol), TBAF (0.8 mL, 0.8 mmol, 1M solution in THF) were added to 6 (450 mg, 0.77 mmol) at room temperature, and the mixture was stirred at room temperature for 5 minutes. A saturated aqueous ammonium chloride solution was added, followed by extraction three times with Et ₂ O. The organic layer was washed with saturated brine, dried over MgSO ₄ , and concentrated under reduced pressure. Thereafter, the product was purified by silica gel column chromatography (n-hexane: EtOAc = 10: 1) to give yellow solid 1- [4- (benzyloxy) phenyl] -3- (2-hydroxyphenyl) propane-1,3- Dione (7) (260 mg, 98%) was obtained.
¹ H NMR (500 MHz, CDCl ₃ ): δ 15.8 (s, 1H), 12.1 (s, 1H), 7.92 (d, J = 9.2 Hz, 2H), 7.77 (dd, J = 8.6, 1.8 Hz, 1H ), 7.50-7.30 (m, 6H), 7.06 (d, J = 9.2 Hz, 2H), 7.00 (dd, J = 8.6, 1.8 Hz, 1H), 6.91 (t, J = 8.6 Hz, 1H), 6.77 (s, 1H), 5.16 (s, 2H)

2-[4-(ベンジルオキシ)フェニル]-4H-クロメン-4-オン(8)の合成
Ar 雰囲気下、0 °C で 7 (300 mg, 0.88 mmol) に THF / MeOH = 5 / 1 (3 mL)、TFA (1.14 mL, 15.3 mmol) を加え、50 °C で 1.5 時間攪拌した。反応液を減圧下濃縮した後、再結晶 (n-ヘキサン: CH₂Cl₂)により精製し、白色個体の 2-[4-(ベンジルオキシ)フェニル]-4H-クロメン-4-オン(8) (180 mg, 62%) を得た。
化合物8 のスペクトルデータは文献に記載のデータとよく一致した（Guz, N. R.; Stermitz, F. R.; Johnson, J. B.; Beeson, T. D.; Willen, S.; Hsiang, J.-F.; Lewis, K. J. Med. Chem. 2001, 44, 261-268）。 Synthesis of 2- [4- (benzyloxy) phenyl] -4H-chromen-4-one (8)
Under Ar atmosphere, THF / MeOH = 5/1 (3 mL) and TFA (1.14 mL, 15.3 mmol) were added to 7 (300 mg, 0.88 mmol) at 0 ° C., and the mixture was stirred at 50 ° C. for 1.5 hours. The reaction mixture was concentrated under reduced pressure and purified by recrystallization (n-hexane: CH ₂ Cl ₂ ) to give white 2- [4- (benzyloxy) phenyl] -4H-chromen-4-one (8) (180 mg, 62%) was obtained.
The spectral data of Compound 8 was in good agreement with the data described in the literature (Guz, NR; Stermitz, FR; Johnson, JB; Beeson, TD; Willen, S .; Hsiang, J.-F .; Lewis, KJ Med. Chem. 2001, 44, 261-268).

2-(4-ヒドロキシフェニル)-4H-クロメン-4-オン(9)の合成
H₂雰囲気下、室温で 8 (100 mg, 0.30 mmol) に EtOH (12 mL)、10% Pd(OH)₂ (5 mg) を加え、60 °C で1 時間攪拌した。反応液をセライトで濾過した後、減圧下濃縮した。その後、再結晶 (Et₂O) により無色個体の2-(4-ヒドロキシフェニル)-4H-クロメン-4-オン(9) (71 mg, 99%) を得た。
¹H NMR (270 MHz, DMSO-d₆): δ 8.02 (dd, J = 8.6, 1.4 Hz, 1H), 7.95 (d, J = 8,6 Hz, 2H), 7.83-7.70 (m, 2H), 7.47 (td, J = 8.6, 1.4 Hz, 1H), 6.93 (d, J = 8.6 Hz, 2H), 6.85 (s, 1H) Synthesis of 2- (4-hydroxyphenyl) -4H-chromen-4-one (9)
EtOH (12 mL) and 10% Pd (OH) ₂ (5 mg) were added to 8 (100 mg, 0.30 mmol) at room temperature under H ₂ atmosphere, and the mixture was stirred at 60 ° C. for 1 hour. The reaction mixture was filtered through celite and concentrated under reduced pressure. Thereafter, recrystallization (Et ₂ O) gave colorless solid 2- (4-hydroxyphenyl) -4H-chromen-4-one (9) (71 mg, 99%).
¹ H NMR (270 MHz, DMSO-d ₆ ): δ 8.02 (dd, J = 8.6, 1.4 Hz, 1H), 7.95 (d, J = 8,6 Hz, 2H), 7.83-7.70 (m, 2H) , 7.47 (td, J = 8.6, 1.4 Hz, 1H), 6.93 (d, J = 8.6 Hz, 2H), 6.85 (s, 1H)

2-[4-(ベンジルオキシ)フェニル]-3-ヒドロキシ-4H-クロメン-4-オン (10)の合成
Ar 雰囲気下、室温で 8 (100 mg, 0.30 mmol) に CH₂Cl₂ (5 mL)、DMDO (9 mL, 0,09-0.11 M in acetone) を加え、30 °C で 2 時間攪拌した。反応液を減圧下濃縮した後、シリカゲルカラムクロマトグラフィー (CH₂Cl₂) により精製し、淡褐色固体の2-[4-(ベンジルオキシ)フェニル]-3-ヒドロキシ-4H-クロメン-4-オン (10) (93.6 mg, 89%) を得た。
化合物10 のスペクトルデータは文献記載のデータとよく一致した（Sobottka, A. M.; Werner, W.; Blaschke, G.; Kiefer, W.; Nowe, U.; Dannhardt, G.; Schapoval, E. E. S.; Schenkel, E. P.; Scriba, G. K. E. Arch. Pharm. (Weinheim, Ger.) 2000, 333, 205-210）。 Synthesis of 2- [4- (benzyloxy) phenyl] -3-hydroxy-4H-chromen-4-one (10)
Under an Ar atmosphere, CH ₂ Cl ₂ (5 mL) and DMDO (9 mL, 0,09-0.11 M in acetone) were added to 8 (100 mg, 0.30 mmol) at room temperature, and the mixture was stirred at 30 ° C. for 2 hr. The reaction mixture was concentrated under reduced pressure and then purified by silica gel column chromatography (CH ₂ Cl ₂ ) to give 2- [4- (benzyloxy) phenyl] -3-hydroxy-4H-chromen-4-one as a light brown solid. (10) (93.6 mg, 89%) was obtained.
The spectral data of compound 10 were in good agreement with the literature data (Sobottka, AM; Werner, W .; Blaschke, G .; Kiefer, W .; Nowe, U .; Dannhardt, G .; Schapoval, EES; Schenkel, EP; Scriba, GKE Arch. Pharm. (Weinheim, Ger.) 2000, 333, 205-210).

3-ヒドロキシ-2-(4-ヒドロキシフェニル)-4H-クロメン-4-オン(11)の合成
H₂雰囲気下、室温で 10 (100 mg, 0.29 mmol) に EtOH (16 mL)、10% Pd(OH)₂ (5 mg) を加え、60 °C で1 時間攪拌した。反応液をセライトで濾過した後、減圧下濃縮した。その後、再結晶 (Et₂O)により淡黄色個体の3-ヒドロキシ-2-(4-ヒドロキシフェニル)-4H-クロメン-4-オン(11) (44 mg, 67%) を得た。
¹H NMR (270 MHz, DMSO-d₆): δ 10.13 (brs, 1H), 9.32 (brs, 1H), 8.10 (d, J = 9.2 Hz, 2H), 8.09 (d, J = 7.9 Hz, 1H), 7.85-7.65 (m ,2H), 7.44 (t, J= 7.9 Hz, 1H), 6.93 (d, J = 9.2 Hz, 2H) Synthesis of 3-hydroxy-2- (4-hydroxyphenyl) -4H-chromen-4-one (11)
EtOH (16 mL) and 10% Pd (OH) ₂ (5 mg) were added to 10 (100 mg, 0.29 mmol) at room temperature under H ₂ atmosphere, and the mixture was stirred at 60 ° C. for 1 hr. The reaction mixture was filtered through celite and concentrated under reduced pressure. Thereafter, 3-hydroxy-2- (4-hydroxyphenyl) -4H-chromen-4-one (11) (44 mg, 67%) as a pale yellow solid was obtained by recrystallization (Et ₂ O).
¹ H NMR (270 MHz, DMSO-d ₆ ): δ 10.13 (brs, 1H), 9.32 (brs, 1H), 8.10 (d, J = 9.2 Hz, 2H), 8.09 (d, J = 7.9 Hz, 1H ), 7.85-7.65 (m, 2H), 7.44 (t, J = 7.9 Hz, 1H), 6.93 (d, J = 9.2 Hz, 2H)

(1H-ベンゾ[d][1,2,3]トリアゾール-1-イル)[3,4-ビス(ベンジルオキシ)フェニル]メタノン(14)の合成
Ar 雰囲気下、0 °C で 12 (11.0 g, 33 mmol) に CH₂Cl₂(105 mL)、SOCl₂(7.2 mL, 99 mmol)、DMF (1.0 mL, 13.2 mmol) を加え、0 °C で 4 時間攪拌した。その後、減圧下濃縮し、無色個体の塩化3,4-ビス(ベンジルオキシ)ベンゾイル(13) を含む粗精製物を得た。続いて、Ar 雰囲気下、0 °C で 13 を含む粗精製物に CH₂Cl₂(150 mL)、Et₃N (4.6 mL, 33 mmol)、1H-ベンゾ[d][1,2,3]トリアゾール (3.9 g, 33 mmol) を加え、0 °C で12 時間攪拌した。飽和塩化アンモニウム水溶液を加えた後、EtOAc で三回抽出し、有機層を 3M 水酸化ナトリウム水溶液で洗浄、MgSO₄ で乾燥、減圧下濃縮した。その後、再結晶 (n-ヘキサン / CH₂Cl₂) により精製し、無色個体の (1H-ベンゾ[d][1,2,3]トリアゾール-1-イル)[3,4-ビス(ベンジルオキシ)フェニル]メタノン(14) (11.4 g, 2 工程、77%) を得た。
¹H NMR (500 MHz, CDCl₃): δ 8.35 (d, J= 8.6 Hz, 1H), 8.17 (d, J = 8.6 Hz, 1H), 7.97 (dd, J = 8.6, 1.8 Hz, 1H), 7.94 (d, J = 2.5 Hz, 1H), 7.69 (td, J = 8.6, 0.95 Hz, 1H), 7.54 (td, J = 8.6, 0.95 Hz, 1H), 7.53-7.45 (m, 4H), 7.43-7.30 (m, 6H), 7.07 (d, J = 8.6 Hz, 1H), 5.22 (s, 2H), 5.19 (s, 2H) Synthesis of (1H-benzo [d] [1,2,3] triazol-1-yl) [3,4-bis (benzyloxy) phenyl] methanone (14)
Add CH ₂ Cl ₂ (105 mL), SOCl ₂ (7.2 mL, 99 mmol), DMF (1.0 mL, 13.2 mmol) to 12 (11.0 g, 33 mmol) at 0 ° C under Ar atmosphere, and add 0 ° C For 4 hours. Thereafter, the mixture was concentrated under reduced pressure to obtain a crude product containing colorless solid 3,4-bis (benzyloxy) benzoyl (13). Subsequently, CH ₂ Cl ₂ (150 mL), Et ₃ N (4.6 mL, 33 mmol), 1H-benzo [d] [1,2,3 were added to a crude product containing 13 at 0 ° C under Ar atmosphere. ] Triazole (3.9 g, 33 mmol) was added, and the mixture was stirred at 0 ° C for 12 hours. A saturated aqueous ammonium chloride solution was added, followed by extraction three times with EtOAc. The organic layer was washed with 3M aqueous sodium hydroxide solution, dried over MgSO ₄ , and concentrated under reduced pressure. After purification by recrystallization (n-hexane / CH ₂ Cl ₂ ), colorless solid (1H-benzo [d] [1,2,3] triazol-1-yl) [3,4-bis (benzyloxy ) Phenyl] methanone (14) (11.4 g, 2 steps, 77%).
¹ H NMR (500 MHz, CDCl ₃ ): δ 8.35 (d, J = 8.6 Hz, 1H), 8.17 (d, J = 8.6 Hz, 1H), 7.97 (dd, J = 8.6, 1.8 Hz, 1H), 7.94 (d, J = 2.5 Hz, 1H), 7.69 (td, J = 8.6, 0.95 Hz, 1H), 7.54 (td, J = 8.6, 0.95 Hz, 1H), 7.53-7.45 (m, 4H), 7.43 -7.30 (m, 6H), 7.07 (d, J = 8.6 Hz, 1H), 5.22 (s, 2H), 5.19 (s, 2H)

1-[3,4-ビス(ベンジルオキシ)フェニル]-3-[2-(tert-ブチルジフェニルシリルオキシ)フェニル]プロパン-1,3-ジオン (15)の合成
Ar 雰囲気下、0 °C で 2 (430 mg, 1.15 mmol) に THF (3.5 mL)、14 (500 mg, 1.15 mmol)、LiHMDS (14 mL, 14 mmol, 1 M溶液、THF中) を加え、室温で 2 時間攪拌した。水を加えた後、EtOAc で三回抽出し、有機層を飽和食塩水で洗浄、MgSO₄ で乾燥、減圧下濃縮した。その後、シリカゲルカラムクロマトグラフィー (n-ヘキサン : EtOAc = 20 : 1 −10 : 1) により精製し、白色固体の 1-[3,4-ビス(ベンジルオキシ)フェニル]-3-[2-(tert-ブチルジフェニルシリルオキシ)フェニル]プロパン-1,3-ジオン (15) (792 mg, quant.) を得た。
¹H NMR (500 MHz, CDCl₃): δ 16.9 (s, 1H), 7.75-7.65 (m, 6H), 7.61 (d, J = 1.9 Hz, 1H), 7.50-7.25 (m, 16 H), 7.00 (td, J = 7.3, 1.9 Hz, 1H), 6.94 (td, J = 7.3, 1.9 Hz, 1H), 6.90 (s, 1H), 6.86 (d, J = 8.5 Hz, 1H), 6.56 (d, J = 7.3 Hz, 1H), 5.22, (s, 2H), 5.13 (s, 2H), 1.01 (s, 9H) Synthesis of 1- [3,4-bis (benzyloxy) phenyl] -3- [2- (tert-butyldiphenylsilyloxy) phenyl] propane-1,3-dione (15)
Under Ar atmosphere, THF (3.5 mL), 14 (500 mg, 1.15 mmol), LiHMDS (14 mL, 14 mmol, 1 M solution in THF) was added to 2 (430 mg, 1.15 mmol) at 0 ° C, Stir at room temperature for 2 hours. Water was added, and the mixture was extracted 3 times with EtOAc. The organic layer was washed with saturated brine, dried over MgSO ₄ , and concentrated under reduced pressure. Then, the product was purified by silica gel column chromatography (n-hexane: EtOAc = 20: 1-10: 1) and 1- [3,4-bis (benzyloxy) phenyl] -3- [2- (tert -Butyldiphenylsilyloxy) phenyl] propane-1,3-dione (15) (792 mg, quant.) Was obtained.
¹ H NMR (500 MHz, CDCl ₃ ): δ 16.9 (s, 1H), 7.75-7.65 (m, 6H), 7.61 (d, J = 1.9 Hz, 1H), 7.50-7.25 (m, 16 H), 7.00 (td, J = 7.3, 1.9 Hz, 1H), 6.94 (td, J = 7.3, 1.9 Hz, 1H), 6.90 (s, 1H), 6.86 (d, J = 8.5 Hz, 1H), 6.56 (d , J = 7.3 Hz, 1H), 5.22, (s, 2H), 5.13 (s, 2H), 1.01 (s, 9H)

1-[3,4-ビス(ベンジルオキシ)フェニル]-3-(2-ヒドロキシフェニル)プロパン-1,3-ジオン (17)の合成
室温で 6 (404 mg, 0.59 mmol) に THF (2 mL)、AcOH (35μL, 0.6 mmol)、TBAF (0.6 mL, 0.6 mmol, 1M溶液、THF中) を加え、室温で 10 分間攪拌した。飽和塩化アンモニウム水溶液を加えた後、EtOAc で三回抽出し、有機層を飽和食塩水で洗浄、MgSO₄で乾燥、減圧下濃縮した。その後、シリカゲルカラムクロマトグラフィー (n-ヘキサン : EtOAc = 10 : 1 to 3 :1) により精製し、黄色個体の 1-[3,4-ビス(ベンジルオキシ)フェニル]-3-(2-ヒドロキシフェニル)プロパン-1,3-ジオン (17) (262 mg, 98%) を得た。
¹H NMR (500 MHz, CDCl₃): δ 15.7 (s, 1H), 12.1 (s, 1H), 7.70 (dd, J = 8.6, 1.9 Hz, 1H), 7.63 (d, J = 1.9 Hz, 1H), 7.55-7.25 (m, 11H), 7.05-6.85 (m, 4H), 6.65 (s, 1H), 5.26 (s, 2H), 5.25 (s, 2H) Synthesis of 1- [3,4-bis (benzyloxy) phenyl] -3- (2-hydroxyphenyl) propane-1,3-dione (17)
THF (2 mL), AcOH (35 μL, 0.6 mmol), TBAF (0.6 mL, 0.6 mmol, 1M solution in THF) were added to 6 (404 mg, 0.59 mmol) at room temperature, and the mixture was stirred at room temperature for 10 minutes. Saturated aqueous ammonium chloride solution was added, and the mixture was extracted 3 times with EtOAc. The organic layer was washed with saturated brine, dried over MgSO ₄ , and concentrated under reduced pressure. Then, it was purified by silica gel column chromatography (n-hexane: EtOAc = 10: 1 to 3: 1), and 1- [3,4-bis (benzyloxy) phenyl] -3- (2-hydroxyphenyl) of a yellow solid ) Propane-1,3-dione (17) (262 mg, 98%) was obtained.
¹ H NMR (500 MHz, CDCl ₃ ): δ 15.7 (s, 1H), 12.1 (s, 1H), 7.70 (dd, J = 8.6, 1.9 Hz, 1H), 7.63 (d, J = 1.9 Hz, 1H ), 7.55-7.25 (m, 11H), 7.05-6.85 (m, 4H), 6.65 (s, 1H), 5.26 (s, 2H), 5.25 (s, 2H)

2-[3,4-ビス(ベンジルオキシ)フェニル]-4H-クロメン-4-オン(18)の合成
Ar 雰囲気下、0 °C で 17 (364 mg, 0.80 mmol) に THF / MeOH = 5 / 1 (3 mL)、TFA (0.7 mL, 9.4 mmol) を加え、50 °C で 1 時間攪拌した。反応液を減圧下濃縮した後、再結晶 (n-ヘキサン: CH₂Cl₂) により精製し、白色個体の 2-[3,4-ビス(ベンジルオキシ)フェニル]-4H-クロメン-4-オン(18) (212 mg, 61%) を得た。¹H NMR (500 MHz, CDCl₃): δ 8.22 (dd, J = 7.9, 1.6 Hz, 1H), 7.68 (td, J = 7.9, 1.6 Hz, 1H), 7.55-7.30 (m, 14H), 7.03 (d, J = 8.5 Hz, 1H), 6.68 (s, 1H), 5.27 (s, 4H) Synthesis of 2- [3,4-bis (benzyloxy) phenyl] -4H-chromen-4-one (18)
Under Ar atmosphere, THF / MeOH = 5/1 (3 mL) and TFA (0.7 mL, 9.4 mmol) were added to 17 (364 mg, 0.80 mmol) at 0 ° C., and the mixture was stirred at 50 ° C. for 1 hour. The reaction mixture was concentrated under reduced pressure and purified by recrystallization (n-hexane: CH ₂ Cl ₂ ) to give 2- [3,4-bis (benzyloxy) phenyl] -4H-chromen-4-one as a white solid. (18) (212 mg, 61%) was obtained. ¹ H NMR (500 MHz, CDCl ₃ ): δ 8.22 (dd, J = 7.9, 1.6 Hz, 1H), 7.68 (td, J = 7.9, 1.6 Hz, 1H), 7.55-7.30 (m, 14H), 7.03 (d, J = 8.5 Hz, 1H), 6.68 (s, 1H), 5.27 (s, 4H)

2-(3,4-ジヒドロキシフェニル)-4H-クロメン-4-オン(19)の合成
H₂雰囲気下、室温で 18 (100 mg, 0.23 mmol) に EtOH (14 mL)、10% Pd(OH)₂ (5 mg) を加え、60 °C で1 時間攪拌した。反応液をセライトで濾過した後、減圧下濃縮した。その後、再結晶 (Et₂O) により淡緑色個体の2-(3,4-ジヒドロキシフェニル)-4H-クロメン-4-オン(19) (36 mg, 62%) を得た。
¹H NMR (270 MHz, DMSO-d₆): δ 8.08 (d, J = 7.9 Hz, 1H), 8.85-7.65 (m, 4H), 7.59 (d, J = 7.3 Hz, 1H), 7.44 (t, J = 7.3 Hz, 1H), 6.89 (d, J = 7.9 Hz, 1H) Synthesis of 2- (3,4-dihydroxyphenyl) -4H-chromen-4-one (19)
EtOH (14 mL) and 10% Pd (OH) ₂ (5 mg) were added to 18 (100 mg, 0.23 mmol) at room temperature under H ₂ atmosphere, and the mixture was stirred at 60 ° C. for 1 hr. The reaction mixture was filtered through celite and concentrated under reduced pressure. Then, recrystallization (Et ₂ O) gave 2- (3,4-dihydroxyphenyl) -4H-chromen-4-one (19) (36 mg, 62%) as a pale green solid.
¹ H NMR (270 MHz, DMSO-d ₆ ): δ 8.08 (d, J = 7.9 Hz, 1H), 8.85-7.65 (m, 4H), 7.59 (d, J = 7.3 Hz, 1H), 7.44 (t , J = 7.3 Hz, 1H), 6.89 (d, J = 7.9 Hz, 1H)

2-[3,4-ビス(ベンジルオキシ)フェニル]-3-ヒドロキシ-4H-クロメン-4-オン (20)の合成
Ar 雰囲気下、-20 °Cで 18 (100 mg, 0.23 mmol) に CH₂Cl₂ (3 mL)、DMDO (7 mL, 0,09-0.11 M、アセトン中) を加え、30 °C で 1 時間攪拌した。反応液を減圧下濃縮した後、シリカゲルカラムクロマトグラフィー (CH₂Cl₂) により精製し、淡褐色固体の 2-[3,4-ビス(ベンジルオキシ)フェニル]-3-ヒドロキシ-4H-クロメン-4-オン (20) (72 mg, 70%) を得た。
化合物20のスペクトルデータは、文献に記載のデータとよく一致した（van Acker, F. A. A.; Hageman, J. A.; Haenen, G. R. M. M.; van der ViJgh, W. J. F.; Best, A.; Menge, W. M. P. B. J. Med. Chem. 2000, 43, 3752-3760）。 Synthesis of 2- [3,4-bis (benzyloxy) phenyl] -3-hydroxy-4H-chromen-4-one (20)
Add CH ₂ Cl ₂ (3 mL) and DMDO (7 mL, 0,09-0.11 M in acetone) to 18 (100 mg, 0.23 mmol) at -20 ° C under Ar atmosphere, and add 1 at 30 ° C. Stir for hours. The reaction mixture was concentrated under reduced pressure and purified by silica gel column chromatography (CH ₂ Cl ₂ ) to give 2- [3,4-bis (benzyloxy) phenyl] -3-hydroxy-4H-chromene- 4-one (20) (72 mg, 70%) was obtained.
The spectral data of compound 20 agreed well with the data described in the literature (van Acker, FAA; Hageman, JA; Haenen, GRMM; van der ViJgh, WJF; Best, A .; Menge, WMPBJ Med. Chem. 2000, 43, 3752-3760).

2-(3,4-ジヒドロキシフェニル)-3-ヒドロキシ-4H-クロメン-4-オン(21)の合成
H₂雰囲気下、室温で 10 (44 mg, 0.097 mmol) に EtOH (5.6 mL)、10% Pd(OH)₂ (3 mg) を加え、60 °C で1 時間攪拌した。反応液をセライトで濾過した後、減圧下濃縮した。その後、再結晶 (Et₂O) により淡緑色個体の2-(3,4-ジヒドロキシフェニル)-3-ヒドロキシ-4H-クロメン-4-オン(21) (20 mg, 82%) を得た。¹H NMR (270 MHz, DMSO-d₆): δ 8.07 (d, J = 7.9 Hz, 1H), 8.85-7.65 (m, 3H), 7.59 (dd, J = 7.9, 2.0 Hz, 1H), 7.44 (t, J = 7.9 Hz, 1H), 6.89 (d, J = 8.6 Hz, 1H) Synthesis of 2- (3,4-dihydroxyphenyl) -3-hydroxy-4H-chromen-4-one (21)
EtOH (5.6 mL) and 10% Pd (OH) ₂ (3 mg) were added to 10 (44 mg, 0.097 mmol) at room temperature under H ₂ atmosphere, and the mixture was stirred at 60 ° C. for 1 hour. The reaction mixture was filtered through celite and concentrated under reduced pressure. Thereafter, recrystallization (Et ₂ O) gave 2- (3,4-dihydroxyphenyl) -3-hydroxy-4H-chromen-4-one (21) (20 mg, 82%) as a pale green solid. ¹ H NMR (270 MHz, DMSO-d ₆ ): δ 8.07 (d, J = 7.9 Hz, 1H), 8.85-7.65 (m, 3H), 7.59 (dd, J = 7.9, 2.0 Hz, 1H), 7.44 (t, J = 7.9 Hz, 1H), 6.89 (d, J = 8.6 Hz, 1H)

(1H-ベンゾ[d][1,2,3]トリアゾール-1-イル)[3,4,5-トリス(ベンジルオキシ)フェニル]メタノン(24)の合成
Ar 雰囲気下、0 °C で 22 (10.0 g, 23 mmol) に CH₂Cl₂(52 mL)、SOCl₂(2.5 mL, 35 mmol)、DMF (0.7 mL, 9.0 mmol) を加え、0 °C で 2 時間攪拌した。その後、減圧下濃縮し、無色個体の塩化ｓ3,4,5-トリス(ベンジルオキシ)ベンゾイル(23) を含む粗精製物を得た。続いて、Ar 雰囲気下、0 °C で 23 を含む粗精製物に CH₂Cl₂(80 mL)、Et₃N (3.2 mL, 23 mmol)、1H-ベンゾ[d][1,2,3]トリアゾール (2.7 g, 23 mmol) を加え、0 °C で24 時間攪拌した。飽和塩化アンモニウム水を加えた後、EtOAc で三回抽出し、有機層を 3M 水酸化ナトリウム水溶液で洗浄、MgSO₄ で乾燥、減圧下濃縮した。その後、再結晶 (n-ヘキサン / CH₂Cl₂) により精製し、無色個体の (1H-ベンゾ[d][1,2,3]トリアゾール-1-イル)[3,4,5-トリス(ベンジルオキシ)フェニル]メタノン(24) (11 g, 2 工程、88%) を得た。
¹H NMR (500 MHz, CDCl₃): δ 8.36 (d, J= 8.3 Hz, 1H), 8.18 (d, J = 8.3 Hz, 1H), 7.71 (td, J = 7.3, 1.0 Hz, 1H), 7.68 (s, 2H), 7.56 (td, J= 7.3, 1.0 Hz, 1H), 7.50-7.20 (m, 15H), 5.20 (s, 2H), 5.19 (s, 4H) Synthesis of (1H-benzo [d] [1,2,3] triazol-1-yl) [3,4,5-tris (benzyloxy) phenyl] methanone (24)
Under Ar atmosphere, add CH ₂ Cl ₂ (52 mL), SOCl ₂ (2.5 mL, 35 mmol), DMF (0.7 mL, 9.0 mmol) to 22 (10.0 g, 23 mmol) at 0 ° C, then add 0 ° C For 2 hours. Thereafter, the reaction mixture was concentrated under reduced pressure to obtain a crude product containing colorless solid s3,4,5-tris (benzyloxy) benzoyl (23). Subsequently, CH ₂ Cl ₂ (80 mL), Et ₃ N (3.2 mL, 23 mmol), 1H-benzo [d] [1,2,3 were added to a crude product containing 23 at 0 ° C under Ar atmosphere. ] Triazole (2.7 g, 23 mmol) was added, and the mixture was stirred at 0 ° C for 24 hours. Saturated aqueous ammonium chloride was added, and the mixture was extracted 3 times with EtOAc. The organic layer was washed with 3M aqueous sodium hydroxide solution, dried over MgSO ₄ , and concentrated under reduced pressure. Then, it was purified by recrystallization (n-hexane / CH ₂ Cl ₂ ), and colorless solid (1H-benzo [d] [1,2,3] triazol-1-yl) [3,4,5-tris ( Benzyloxy) phenyl] methanone (24) (11 g, 2 steps, 88%) was obtained.
¹ H NMR (500 MHz, CDCl ₃ ): δ 8.36 (d, J = 8.3 Hz, 1H), 8.18 (d, J = 8.3 Hz, 1H), 7.71 (td, J = 7.3, 1.0 Hz, 1H), 7.68 (s, 2H), 7.56 (td, J = 7.3, 1.0 Hz, 1H), 7.50-7.20 (m, 15H), 5.20 (s, 2H), 5.19 (s, 4H)

1-[2-(tert-ブチルジフェニルシリルオキシ)フェニル]-3-[3,4,5-トリス(ベンジルオキシ)フェニル]プロパン-1,3-ジオン (25)の合成
Ar 雰囲気下、0 °Cで 2 (1.0 g, 2.7 mmol) に THF (6.0 mL)、24 (1.73 g, 3.19 mmol)、LiHMDS (11 mL, 11 mmol, 1 M溶液、THF中) を加え、室温で 3.5 時間攪拌した。飽和塩化アンモニウム水溶液を加えた後、CH₂Cl₂ で三回抽出し、有機層を飽和食塩水で洗浄、MgSO₄ で乾燥、減圧下濃縮した。その後、シリカゲルカラムクロマトグラフィー (n-ヘキサン: EtOAc = 20 : 1 − 10 : 1) により精製し、白色固体の 1-[2-(tert-ブチルジフェニルシリルオキシ)フェニル]-3-[3,4,5-トリス(ベンジルオキシ)フェニル]プロパン-1,3-ジオン (25) (1.83 g, 86%) を得た。
¹H NMR (500 MHz, CDCl₃): δ 16.8 (s, 1H), 7.72 (dd, J = 7.9, 1.2 Hz, 4H), 7.69 (dd, J = 7.3, 1.8 Hz, 1H), 7.45-7.27 (m, 21H), 7.25 (s, 2H), 7.04 (td, J = 7.3, 1.8 Hz, 1H), 6.97 (td, J = 7.3, 1.8 Hz, 1H), 6.89 (s, 1H), 6.57 (dd, J = 7.3, 1.8 Hz, 1H), 5.12 (s, 2H), 5.01 (s, 4H), 1.02 (s, 9H) Synthesis of 1- [2- (tert-butyldiphenylsilyloxy) phenyl] -3- [3,4,5-tris (benzyloxy) phenyl] propane-1,3-dione (25)
Under Ar atmosphere, THF (6.0 mL), 24 (1.73 g, 3.19 mmol), LiHMDS (11 mL, 11 mmol, 1 M solution in THF) was added to 2 (1.0 g, 2.7 mmol) at 0 ° C, Stir at room temperature for 3.5 hours. A saturated aqueous ammonium chloride solution was added, and the mixture was extracted 3 times with CH ₂ Cl _{2. The} organic layer was washed with saturated brine, dried over MgSO ₄ , and concentrated under reduced pressure. Thereafter, the residue was purified by silica gel column chromatography (n-hexane: EtOAc = 20: 1-10: 1) and 1- [2- (tert-butyldiphenylsilyloxy) phenyl] -3- [3,4 , 5-tris (benzyloxy) phenyl] propane-1,3-dione (25) (1.83 g, 86%) was obtained.
¹ H NMR (500 MHz, CDCl ₃ ): δ 16.8 (s, 1H), 7.72 (dd, J = 7.9, 1.2 Hz, 4H), 7.69 (dd, J = 7.3, 1.8 Hz, 1H), 7.45-7.27 (m, 21H), 7.25 (s, 2H), 7.04 (td, J = 7.3, 1.8 Hz, 1H), 6.97 (td, J = 7.3, 1.8 Hz, 1H), 6.89 (s, 1H), 6.57 ( dd, J = 7.3, 1.8 Hz, 1H), 5.12 (s, 2H), 5.01 (s, 4H), 1.02 (s, 9H)

1-(2-ヒドロキシフェニル)-3-[3,4,5-トリス(ベンジルオキシ)フェニル]プロパン-1,3-ジオン (26)の合成
室温で 6 (1.7 g, 2.1 mmol) に THF (17 mL)、AcOH (0.12 mL, 2.1 mmol)、TBAF (2.2 mL, 2.2 mmol, 1M 溶液、THF) を加え、室温で 10 分間攪拌した。飽和塩化アンモニウム水溶液を加えた後、EtOAc で三回抽出し、有機層を飽和食塩水で洗浄、MgSO₄で乾燥、減圧下濃縮した。その後、シリカゲルカラムクロマトグラフィー (n-ヘキサン : EtOAc = 10 : 1 to 5 :1) により精製し、黄色個体の 1-(2-ヒドロキシフェニル)-3-[3,4,5-トリス(ベンジルオキシ)フェニル]プロパン-1,3-ジオン (26) (998 mg, 85%) を得た。
¹H NMR (500 MHz, CDCl₃): δ 15.6 (s, 1H), 12.0 (s, 1H), 7.67 (dd, J = 7.9, 1.2 Hz, 1H), 7.50-7.22 (m, 16H), 7.21 (s, 2H), 7.00 (d, J= 7.9 Hz, 1H), 6.94 (td, J = 7.9, 1.2 Hz, 1H), 6.57 (s, 1H), 5.19 (s, 4H), 5.16 (s, 2H) Synthesis of 1- (2-hydroxyphenyl) -3- [3,4,5-tris (benzyloxy) phenyl] propane-1,3-dione (26)
THF (17 mL), AcOH (0.12 mL, 2.1 mmol) and TBAF (2.2 mL, 2.2 mmol, 1M solution, THF) were added to 6 (1.7 g, 2.1 mmol) at room temperature, and the mixture was stirred at room temperature for 10 minutes. Saturated aqueous ammonium chloride solution was added, and the mixture was extracted 3 times with EtOAc. The organic layer was washed with saturated brine, dried over MgSO ₄ , and concentrated under reduced pressure. Then, it was purified by silica gel column chromatography (n-hexane: EtOAc = 10: 1 to 5: 1) to give yellow solid 1- (2-hydroxyphenyl) -3- [3,4,5-tris (benzyloxy ) Phenyl] propane-1,3-dione (26) (998 mg, 85%) was obtained.
¹ H NMR (500 MHz, CDCl ₃ ): δ 15.6 (s, 1H), 12.0 (s, 1H), 7.67 (dd, J = 7.9, 1.2 Hz, 1H), 7.50-7.22 (m, 16H), 7.21 (s, 2H), 7.00 (d, J = 7.9 Hz, 1H), 6.94 (td, J = 7.9, 1.2 Hz, 1H), 6.57 (s, 1H), 5.19 (s, 4H), 5.16 (s, 2H)

2-[3,4,5-トリス(ベンジルオキシ)フェニル]-4H-クロメン-4-オン (27)の合成
Ar 雰囲気下、0 °C で 7 (610 mg, 1.09 mmol) に THF / MeOH = 5 / 1 (3.6 mL)、TFA (2.4 mL, 8.5 mmol) を加え、50 °C で 3 時間攪拌した。反応液を減圧下濃縮した後、シリカゲルカラムクロマトグラフィー (n-ヘキサン : CH₂Cl₂= 5 :1) により精製し、無色固体の 2-[3,4,5-トリス(ベンジルオキシ)フェニル]-4H-クロメン-4-オン (27) (466 mg, 79%) を得た。
¹H NMR (500 MHz, CDCl₃): δ 8.22 (dd, J= 7.9, 1.6 Hz, 1H), 7.70 (td, J= 7.9, 1.6 Hz, 1H), 7.53 (d, J= 7.9 Hz, 1H), 7.50-7.25 (m, 16H), 7.21 (s, 2H), 6.68 (s, 1H), 5.20 (s, 4H), 5.16 (s, 2H) Synthesis of 2- [3,4,5-tris (benzyloxy) phenyl] -4H-chromen-4-one (27)
Under Ar atmosphere, THF / MeOH = 5/1 (3.6 mL) and TFA (2.4 mL, 8.5 mmol) were added to 7 (610 mg, 1.09 mmol) at 0 ° C., and the mixture was stirred at 50 ° C. for 3 hours. The reaction solution was concentrated under reduced pressure and purified by silica gel column chromatography (n-hexane: CH ₂ Cl ₂ = 5: 1) to give 2- [3,4,5-tris (benzyloxy) phenyl] as a colorless solid. -4H-chromen-4-one (27) (466 mg, 79%) was obtained.
¹ H NMR (500 MHz, CDCl ₃ ): δ 8.22 (dd, J = 7.9, 1.6 Hz, 1H), 7.70 (td, J = 7.9, 1.6 Hz, 1H), 7.53 (d, J = 7.9 Hz, 1H ), 7.50-7.25 (m, 16H), 7.21 (s, 2H), 6.68 (s, 1H), 5.20 (s, 4H), 5.16 (s, 2H)

2-(3,4,5-トリヒドロキシフェニル)-4H-クロメン-4-オン(28)の合成
H₂雰囲気下、室温で 27 (100 mg, 0.18 mmol) に EtOH (4 mL)、10% Pd(OH)₂ (9 mg) を加え、60 °C で2.5 時間攪拌した。反応液をセライトで濾過した後、減圧下濃縮した。その後、再結晶 (Et₂O) により緑色個体の2-(3,4,5-トリヒドロキシフェニル)-4H-クロメン-4-オン(28) (26 mg, 54%) を得た。
¹H NMR (270 MHz, DMSO-d₆): δ 8.08 (dd, J = 7.9, 1.3 Hz, 1H), 7.77 (td, J = 7.9, 1.3 Hz, 1H), 7.66 (d, J = 7.9 Hz, 1H), 7.44 (t, J = 7.9 Hz, 1H), 7.29 (s, 2H), 6.89 (s, 1H) Synthesis of 2- (3,4,5-trihydroxyphenyl) -4H-chromen-4-one (28)
EtOH (4 mL) and 10% Pd (OH) ₂ (9 mg) were added to 27 (100 mg, 0.18 mmol) at room temperature under H ₂ atmosphere, and the mixture was stirred at 60 ° C. for 2.5 hours. The reaction mixture was filtered through celite and concentrated under reduced pressure. Then, 2- (3,4,5-trihydroxyphenyl) -4H-chromen-4-one (28) (26 mg, 54%) as a green solid was obtained by recrystallization (Et ₂ O).
¹ H NMR (270 MHz, DMSO-d ₆ ): δ 8.08 (dd, J = 7.9, 1.3 Hz, 1H), 7.77 (td, J = 7.9, 1.3 Hz, 1H), 7.66 (d, J = 7.9 Hz , 1H), 7.44 (t, J = 7.9 Hz, 1H), 7.29 (s, 2H), 6.89 (s, 1H)

3-ヒドロキシ-2-(3,4,5-トリヒドロキシフェニル)-4H-クロメン-4-オン(30)の合成
Ar 雰囲気下、0 ℃で 27 (300 mg, 0.555 mmol) に CH₂Cl₂ (30 mL)、DMDO (60 mL, 0,09-0.11 M、アセトン中) を加え、室温で 2 時間攪拌した。反応液を減圧下濃縮した後、淡褐色固体の 3-ヒドロキシ-2-[3,4,5-トリス(ベンジルオキシ)フェニル]-4H-クロメン-4-オン(29) を含む粗精製物を得た。
続いて、H₂雰囲気下、室温で 29 を含む粗精製物にEtOH (23 mL)、10% Pd(OH)₂ (23 mg) を加え、70 ℃で 1 時間攪拌した。反応液をセライトで濾過した後、減圧下濃縮した。その後、再結晶 (MeOH) により緑色個体の3-ヒドロキシ-2-(3,4,5-トリヒドロキシフェニル)-4H-クロメン-4-オン(30) (83.5 mg, 60%) を得た。
¹H NMR (270 MHz, DMSO-d₆): δ 8.15 (dd, J = 7.9, 1.3 Hz, 1H), 7.84 (td, J = 7.9, 1.3 Hz, 1H), 7.73 (d, J = 7.9 Hz, 1H), 7.51 (t, J = 7.9 Hz, 1H), 7.37 (s, 2H), 6.97 (s, 1H) Synthesis of 3-hydroxy-2- (3,4,5-trihydroxyphenyl) -4H-chromen-4-one (30)
Under Ar atmosphere, CH ₂ Cl ₂ (30 mL) and DMDO (60 mL, 0,09-0.11 M in acetone) were added to 27 (300 mg, 0.555 mmol) at 0 ° C., and the mixture was stirred at room temperature for 2 hours. After the reaction solution was concentrated under reduced pressure, a crude product containing 3-hydroxy-2- [3,4,5-tris (benzyloxy) phenyl] -4H-chromen-4-one (29) as a light brown solid was obtained. Obtained.
Subsequently, EtOH (23 mL) and 10% Pd (OH) ₂ (23 mg) were added to the crude product containing 29 at room temperature under H ₂ atmosphere, and the mixture was stirred at 70 ° C. for 1 hour. The reaction mixture was filtered through celite and concentrated under reduced pressure. Thereafter, 3-hydroxy-2- (3,4,5-trihydroxyphenyl) -4H-chromen-4-one (30) (83.5 mg, 60%) as a green solid was obtained by recrystallization (MeOH).
¹ H NMR (270 MHz, DMSO-d ₆ ): δ 8.15 (dd, J = 7.9, 1.3 Hz, 1H), 7.84 (td, J = 7.9, 1.3 Hz, 1H), 7.73 (d, J = 7.9 Hz , 1H), 7.51 (t, J = 7.9 Hz, 1H), 7.37 (s, 2H), 6.97 (s, 1H)

実施例２フラボン誘導体のシアル酸転移酵素阻害活性試験
フラボン誘導体のシアル酸転移酵素阻害活性を以下の方法で測定した。シアル酸転移酵素としては、組換え体ヒト由来シアル酸転移酵素（hST6Gal I）、およびラット由来シアル酸転移酵素（rST6Gal I、Calbiochem）を用いた。Universal bind, 1x8 stripwell^TM プレート (COSTAR（登録商標）,#29306009) に0.005 μMのLacNAc-5AP-γPGA含有PBS溶液を35 μl/ウエル加え、遮光した状態で37℃で一晩インキュベーションした後、UV254 nmを1分間照射し固層化した。未結合の溶液を除去し、2% (w/v) BSA含有PBS溶液を300 μl/ウエル加え、3時間以上, 4℃でブロッキングした。BSA含有PBS溶液を除去し、0.05% (w/v) Tween20含有PBS溶液を300 μl/ウエル加え洗浄した。同様の洗浄を3回繰り返した。プレートを氷上に置き、最終濃度がhST6Gal Iについては [3 micro U hST6Gal I, 50 mM カコジル酸ナトリウムバッファー(pH 6.5) , 2 mM MnCl₂, 0.5% (w/v) Triton CF-54, 0-15 μM CMP-Neu5Ac] に、rST6Gal Iについては [10 micro U rST6Gal I, 50 mM カコジル酸ナトリウムバッファー(pH 6.5) , 2 mM MnCl₂, 0.5% (w/v) Triton CF-54, 0-15 μM CMP-Neu5Ac]になるようにそれぞれ調製し、適宜希釈した試験化合物を加えて、50 μl/ウエル, 1時間, 37℃で酵素反応させた。反応溶液を除去し、0.05% (w/v) Tween20含有PBS溶液を300 μl/ウエル加え洗浄した。同様の洗浄を3回繰り返した。1ｍg/mlビオチン化Sambucus sieboldiana アグルチニン (SSA) (J-オイルミルズ, #300442) 含有PBS溶液を0.02% (w/v) Tween20含有PBS溶液で1,000倍希釈し、50 μl/ウエル, 1時間, 室温で反応させた。レクチン溶液を除去し、0.05% (w/v) Tween20含有PBS溶液を300 μl/ウエル加え洗浄した。同様の洗浄を3回繰り返した。アルカリホスファターゼ(AP) 標識ストレプトアビジン(Promega, V559C #18181908) を0.02% (w/v) Tween20含有PBS溶液で10,000倍希釈し、50 μl/ウエル, 1時間, 室温で反応させた。ストレプトアビジン-AP溶液を除去し、0.05% (w/v) Tween20含有PBS溶液を300 μl/ウエル加え洗浄した。同様の洗浄を3回繰り返した。基質溶液 [1 mg/ml p-ニトロフェニルリン酸二ナトリウム, 50 mM MgCl₂, 100mM ジエタノールアミンバッファー(pH 9.8) ] を100 μl/ウエル, 30分間, 37℃で反応させた後415 nm における吸光度を測定した (対照492 nm)。結果を図１および図２に示す。 Example 2 Sialyltransferase inhibitory activity test of flavone derivatives Sialyltransferase inhibitory activity of flavone derivatives was measured by the following method. As the sialyltransferase, recombinant human-derived sialyltransferase (hST6Gal I) and rat-derived sialyltransferase (rST6Gal I, Calbiochem) were used. Universal bind, 1x8 stripwell ^TM plate (COSTAR®, The PBS solution containing 0.005 μM LacNAc-5AP-γPGA was added to # 29306009) and incubated overnight at 37 ° C. in a light-shielded state, and then irradiated to UV254 nm for 1 minute to solidify. Unbound solution was removed, PBS solution containing 2% (w / v) BSA was added at 300 μl / well, and blocked at 4 ° C. for 3 hours or more. The BSA-containing PBS solution was removed, and a PBS solution containing 0.05% (w / v) Tween 20 was added at 300 μl / well for washing. The same washing was repeated 3 times. Place the plate on ice for a final concentration of hST6Gal I (3 micro U hST6Gal I, 50 mM sodium cacodylate buffer (pH 6.5), 2 mM MnCl ₂ , 0.5% (w / v) Triton CF-54, 0- 15 μM CMP-Neu5Ac] for rST6Gal I [10 micro U rST6Gal I, 50 mM sodium cacodylate buffer (pH 6.5), 2 mM MnCl ₂ , 0.5% (w / v) Triton CF-54, 0-15 [MuM CMP-Neu5Ac] was prepared, and each test compound diluted appropriately was added, followed by enzyme reaction at 50 μl / well for 1 hour at 37 ° C. The reaction solution was removed, and a PBS solution containing 0.05% (w / v) Tween 20 was added and washed with 300 μl / well. The same washing was repeated 3 times. 1 mg / ml biotinylated Sambucus sieboldiana Agglutinin (SSA) (J-Oil Mills, # 300442) PBS solution containing 0.02% (w / v) Tween20 in PBS diluted 1,000 times, 50 μl / well, 1 hour, room temperature It was made to react with. The lectin solution was removed, and a PBS solution containing 0.05% (w / v) Tween 20 was added and washed with 300 μl / well. The same washing was repeated 3 times. Alkaline phosphatase (AP) -labeled streptavidin (Promega, V559C # 18181908) was diluted 10,000-fold with a PBS solution containing 0.02% (w / v) Tween 20 and allowed to react at room temperature for 50 μl / well for 1 hour. The streptavidin-AP solution was removed, and a PBS solution containing 0.05% (w / v) Tween 20 was added and washed with 300 μl / well. The same washing was repeated 3 times. Substrate solution [1 mg / ml p-nitrophenyl phosphate disodium, 50 mM MgCl ₂ , 100 mM diethanolamine buffer (pH 9.8)] was reacted at 100 μl / well for 30 minutes at 37 ° C, and the absorbance at 415 nm was measured. Measured (control 492 nm). The results are shown in FIG. 1 and FIG.

図に示されるように、フラボン誘導体はシアル酸転移酵素阻害活性を有していた。最も強い活性を示したトリデオキシ体は、天然に存在しないだけでなく、これまでに報告がない化合物である。これらのシアル酸転移酵素阻害活性を示す化合物は、生体内でシアル酸が関与する疾患（炎症、がん転移、ウイルス感染症）に対して有効な治療薬となることが期待できる化合物である。 As shown in the figure, the flavone derivative had sialyltransferase inhibitory activity. The trideoxy form which showed the strongest activity is a compound which not only does not exist naturally but has not been reported so far. These compounds exhibiting sialic acid transferase inhibitory activity are compounds that can be expected to be effective therapeutic agents for diseases involving sialic acid in vivo (inflammation, cancer metastasis, viral infection).

図１は、本発明のフラボン誘導体のシアル酸転移酵素阻害活性を示す。FIG. 1 shows the sialyltransferase inhibitory activity of the flavone derivative of the present invention. 図２は、本発明のフラボン誘導体のシアル酸転移酵素阻害活性を示す。FIG. 2 shows the sialyltransferase inhibitory activity of the flavone derivative of the present invention.

Claims

Formula (I):
[Wherein R ₁ is H or OH, and n is 1, 2 or 3]
A method for producing a compound represented by
(A) The following formula (II):
Protecting the hydroxyl group of the compound represented by the following formula (III):
[Wherein R ₂ is a hydroxyl-protecting group]
A compound represented by:
(B) The following formula (IV):
[Wherein n is as defined above]
A protective group is introduced into the compound represented by the following formula (V):
[Wherein R ₃ is a protecting group for a hydroxyl group, R ₄ is benzotriazole which is a protecting group for an acyl group, and n is as defined above]
A compound represented by:
(C) reacting a compound of formula (III) with a compound of formula (V) to give the following formula (VI):
[Wherein R ₂ , R ₃ and n are as defined above]
A compound represented by:
(D) removing the protecting group R ₂ of the compound of formula (VI) to give the following formula (VII):
[Wherein R ₃ and n are as defined above]
A compound represented by:
(E) The compound of formula (VII) is cyclized to give the following formula (VIII):
[Wherein R ₃ and n are as defined above]
A compound represented by:
(F) optionally oxidizing the 3-position of the chromone ring of the compound of formula (VIII) to give the following formula (VIII ′):
[Wherein R ₃ and n are as defined above]
A compound represented by:
(G) removing the protecting group for the hydroxyl group of the compound of formula (VIII) or (VIII ′) to give a compound of the general formula (I):
To obtain a compound of
The method including each process of these.

The compound of general formula (I) is one of the following compounds:
The method of claim 1, wherein