JP2024068197A

JP2024068197A - Ionizable perfluorophosphoryl crown ethers.

Info

Publication number: JP2024068197A
Application number: JP2023189314A
Authority: JP
Inventors: 卓三相田; アビールゴスワミ; フービャオファン
Original assignee: RIKEN Institute of Physical and Chemical Research
Current assignee: RIKEN Institute of Physical and Chemical Research
Priority date: 2022-11-07
Filing date: 2023-11-06
Publication date: 2024-05-17

Abstract

【課題】水溶性であり、凝集などなく水中で安定に存在し、かつ多様な機能性基を導入可能なＦリッチな有機フッ素化合物を提供する。【解決手段】次式（Ｉ）：【化１】TIFF2024068197000021.tif49117（式中、Ｒは水酸基、塩素原子又は式：－Ｏ－（有機基）、－ＮＨ－（有機基）もしくは－Ｓ－（有機基）で示される基を表し、ｎは１～７の整数を表す。）で示される化合物、当該化合物の製造方法、及び当該化合物と水系溶媒とを含む組成物。【選択図】なし[Problem] To provide an F-rich organic fluorine compound that is water-soluble, exists stably in water without aggregation, and allows introduction of various functional groups. [Solution] A compound represented by the following formula (I): [Chemical 1] TIFF2024068197000021.tif49117 (wherein R represents a hydroxyl group, a chlorine atom, or a group represented by the formula: -O- (organic group), -NH- (organic group) or -S- (organic group), and n represents an integer of 1 to 7), a method for producing said compound, and a composition containing said compound and an aqueous solvent. [Selected Figure] None

Description

本発明は、イオン化可能なパーフルオロホスホリルクラウンエーテルに関する。 The present invention relates to ionizable perfluorophosphoryl crown ethers.

パーフルオロ有機化合物は生体内には存在しないが、そのユニークな性質から医薬品や農薬に広く利用されている。フッ素化は、高い血清安定性や生体分子の細胞透過性、非侵襲的なイメージング能力をもたらす。現在の医薬品の４分の１は、少なくとも１つのフッ素原子を含んでいる。 Perfluorinated organic compounds are not found in living organisms, but their unique properties make them widely used in pharmaceuticals and agrochemicals. Fluorination provides high serum stability, cell permeability of biomolecules, and non-invasive imaging capabilities. One-quarter of current pharmaceuticals contain at least one fluorine atom.

プロトンＭＲＩは、非侵襲的な生体イメージング技術として一般的に利用されている。しかし、従来のプロトンＭＲＩで使われるガドリニウム造影剤は、脳内に滞留する性質に起因して副作用を引き起こし、毒性が強い。近年では、ターゲットからの信号を直接検出でき、かつＳ／Ｎ比を向上させることができる^１９ＦＭＲＩがプロトンＭＲＩの代替技術として大きな関心を集めている。例えば、特許文献１には、パーフルオロ－ｔ－ブチルシクロヘキサンを分子イメージング用造影剤として使用できることが記載されている。しかし、パーフルオロ化合物は、水への溶解性が低く、化学的に不活性であり、水や土壌資源を汚染するだけでなく、生体内の様々な部位に蓄積するため、その長期間の使用や大量の使用には問題があることが最近の研究により明らかになってきた。 Proton MRI is commonly used as a non-invasive biological imaging technique. However, the gadolinium contrast agent used in conventional proton MRI causes side effects due to its tendency to remain in the brain and is highly toxic. In recent years, ¹⁹ F MRI, which can directly detect signals from targets and improve the S/N ratio, has attracted great interest as an alternative technique to proton MRI. For example, Patent Document 1 describes that perfluoro-t-butylcyclohexane can be used as a contrast agent for molecular imaging. However, recent research has revealed that perfluoro compounds have low solubility in water, are chemically inactive, and not only pollute water and soil resources but also accumulate in various parts of the body, so that their long-term use or large-scale use is problematic.

そこで、例えば非特許文献１では、パーフルオロオクチルブロマイドなどのパーフルオロ化合物のナノエマルジョンを安定化させるため手段として、標的リガンド（例えば、抗体、ペプチドなど）と共に界面活性分子（乳化剤）を混合することが提案された。特許文献２には、パーフルオロヘキシルオクタン、パーフルオロブチルペンタン等のセミフッ化化合物、当該セミフッ化化合物と混和可能であるトリグリセリド、及び乳化剤を含む水性エマルジョンが造影剤として使用できることが記載されている。しかしながら、安定な水性エマルジョンを形成するために、より水に溶けやすく、より容易に官能基を付加しやすいＦリッチな化合物を設計する新しい戦略を見つけることは依然として挑戦的な課題である。 For example, Non-Patent Document 1 proposes mixing a surface active molecule (emulsifier) with a target ligand (e.g., antibody, peptide, etc.) as a means for stabilizing nanoemulsions of perfluoro compounds such as perfluorooctyl bromide. Patent Document 2 describes that an aqueous emulsion containing a semifluorinated compound such as perfluorohexyl octane or perfluorobutyl pentane, a triglyceride miscible with the semifluorinated compound, and an emulsifier can be used as a contrast agent. However, it remains a challenging task to find a new strategy for designing F-rich compounds that are more soluble in water and easier to add functional groups to in order to form stable aqueous emulsions.

国際公開第２０１１／０１３０３８号International Publication No. 2011/013038 国際公開第２０１４／１５４５３１号International Publication No. 2014/154531

I. Tirotta, et al., Chem. Rev. 2015, 115, 1106I. Tirotta, et al., Chem. Rev. 2015, 115, 1106

本発明は、前記従来技術の有する課題に鑑みてなされたものであり、水溶性であり、凝集などなく水中で安定に存在し、かつ多様な機能性基を導入可能なＦリッチな有機フッ素化合物を提供することを目的とする。 The present invention has been made in consideration of the problems associated with the above-mentioned conventional techniques, and aims to provide an F-rich organic fluorine compound that is water-soluble, exists stably in water without aggregation, and can introduce a variety of functional groups.

本発明者らは、前記課題を解決すべく鋭意研究を行った結果、フッ素化ポリエチレングリコールを塩化ホスホリルと反応させて、塩素化されたパーフルオロホスホリルクラウンエーテルを得た後、水と反応させたところ、高収率で、イオン化可能なパーフルオロホスホリルクラウンエーテル（Proton-Ionizable Perfluorinated Phosphoryl Crown Ether）（以下「ＰＩＰＰＥＲ」という。）が得られることを見出した。また得られた中間体の塩素化されたパーフルオロホスホリルクラウンエーテル（以下「ＰＩＰＰＥＲ－Ｃｌ」という。）は、反応性に富み、求核付加－脱離反応により、マレイミド基、アルキン基、１級アミノ基、グアニジニウム基、蛍光体、薬剤などの任意の官能基を一段階で高収率で導入できることを見出し、本発明を完成するに至った。 The inventors conducted intensive research to solve the above problems and discovered that by reacting fluorinated polyethylene glycol with phosphoryl chloride to obtain a chlorinated perfluorophosphoryl crown ether, which was then reacted with water, a proton-ionizable perfluorinated phosphoryl crown ether (hereinafter referred to as "PIPPER") was obtained in high yield. The inventors also discovered that the intermediate chlorinated perfluorophosphoryl crown ether (hereinafter referred to as "PIPPER-Cl") obtained is highly reactive, and that any functional group, such as a maleimide group, an alkyne group, a primary amino group, a guanidinium group, a fluorescent material, or a drug, can be introduced in one step in high yield by a nucleophilic addition-elimination reaction, which led to the completion of the present invention.

すなわち、本発明の要旨は以下のとおりである。
（１）次式（Ｉ）：

（式中、Ｒは水酸基、塩素原子又は式：－Ｏ－（有機基）、－ＮＨ－（有機基）もしくは－Ｓ－（有機基）で示される基を表し、ｎは１～７の整数を表す。）
で示される化合物。
（２）前記式（Ｉ）において、Ｒが塩素原子である前記（１）に記載の化合物。
（３）前記式（Ｉ）において、Ｒが水酸基である前記（１）に記載の化合物。
（４）前記式（Ｉ）において、Ｒが式：－Ｏ－（有機基）、－ＮＨ－（有機基）又は－Ｓ－（有機基）で示される基であり、前記有機基がアルキン基又はマレイミド基を少なくとも１つ含む前記（１）に記載の化合物。
（５）前記式（Ｉ）において、Ｒが式：－Ｏ－（有機基）、－ＮＨ－（有機基）又は－Ｓ－（有機基）で示される基であり、前記有機基が、医薬もしくは診断薬の有効成分に由来する基を含む前記（１）に記載の化合物。
（６）前記式（Ｉ）において、Ｒが式：－Ｏ－（有機基）、－ＮＨ－（有機基）又は－Ｓ－（有機基）で示される基であり、前記有機基が、発光性及び／又は光吸収性の機能性基を含む前記（１）に記載の化合物。 That is, the gist of the present invention is as follows.
(1) The following formula (I):

(In the formula, R represents a hydroxyl group, a chlorine atom, or a group represented by the formula: -O- (organic group), -NH- (organic group), or -S- (organic group), and n represents an integer of 1 to 7.)
A compound represented by the formula:
(2) The compound according to (1) above, wherein in formula (I), R is a chlorine atom.
(3) The compound according to (1) above, wherein in formula (I), R is a hydroxyl group.
(4) The compound according to (1), wherein in formula (I), R is a group represented by the formula: -O- (organic group), -NH- (organic group), or -S- (organic group), and the organic group contains at least one alkyne group or maleimide group.
(5) The compound according to (1), wherein in formula (I), R is a group represented by the formula: -O- (organic group), -NH- (organic group), or -S- (organic group), and the organic group contains a group derived from an active ingredient of a pharmaceutical or diagnostic agent.
(6) The compound according to (1), wherein in formula (I), R is a group represented by the formula: -O- (organic group), -NH- (organic group), or -S- (organic group), and the organic group contains a light-emitting and/or light-absorbing functional group.

（７）次式（II）：
ＨＯＣＨ_２ＣＦ_２Ｏ－（ＣＦ_２ＣＦ_２Ｏ）_ｎ－ＣＦ_２ＣＨ_２ＯＨ（II）
（式中、ｎは請求項１記載の前記式（Ｉ）と同義である。）
で示されるフッ素化ポリエチレングリコールを塩化ホスホリルと反応させることを含む、前記（２）に記載の化合物の製造方法。
（８）前記（２）に記載の化合物を水と反応させることを含む、前記（３）に記載の化合物の製造方法。
（９）前記（２）に記載の化合物を式：ＨＯ－（有機基）、Ｈ_２Ｎ－（有機基）又はＨＳ－（有機基）で示される化合物と反応させることを含む、前記式（Ｉ）において、Ｒが式：－Ｏ－（有機基）、－ＮＨ－（有機基）又は－Ｓ－（有機基）で示される基である化合物の製造方法。
（１０）前記（４）に記載の化合物と、反応試薬とをクリック反応させることを含む前記（５）又は（６）に記載の化合物の製造方法。
（１１）前記（１）～（６）のいずれかに記載の化合物と、水系溶媒とを含む組成物。
（１２）更に無機イオンを含む前記（１１）に記載の組成物。
（１３）前記（１）～（６）のいずれかに記載の化合物を含む無機イオン包接剤。 (7) The following formula (II):
_HOCH2CF2O- ₍ _CF2CF2O ) _n- _CF2CH2OH ₍ _II )
(In the formula, n has the same meaning as in formula (I) described in claim 1.)
A method for producing the compound according to (2) above, comprising reacting a fluorinated polyethylene glycol represented by the following formula with phosphoryl chloride.
(8) A method for producing the compound according to (3) above, comprising reacting the compound according to (2) above with water.
(9) A method for producing a compound of formula (I) in which R is a group represented by the formula: -O- (organic group), _-NH- (organic group) or -S- (organic group), comprising reacting the compound according to (2) above with a compound represented by the formula: HO- (organic group), H 2 N- (organic group) or HS- (organic group).
(10) A method for producing the compound according to (5) or (6) above, comprising subjecting the compound according to (4) above to a click reaction with a reaction reagent.
(11) A composition comprising the compound according to any one of (1) to (6) above and an aqueous solvent.
(12) The composition according to (11) above, further comprising an inorganic ion.
(13) An inorganic ion inclusion agent comprising the compound according to any one of (1) to (6) above.

本発明によれば、水溶性であり、凝集などなく水中で安定に存在し、かつ多様な機能性基を導入可能なＦリッチな有機フッ素化合物を提供することができる。 The present invention provides an F-rich organic fluorine compound that is water-soluble, exists stably in water without aggregation, and can introduce a variety of functional groups.

以下、本発明を詳細に説明する。 The present invention is described in detail below.

前記式（Ｉ）において、Ｒで表される式：－Ｏ－（有機基）、－ＮＨ－（有機基）又は－Ｓ－（有機基）で示される基における有機基としては、医薬の有効成分に由来する基、発光性基及び／又は光吸収性基（これらの基は診断薬の有効成分に由来する基であってもよい）、又は官能基（例えば反応試薬として有用なマレイミド基、アルキン基）を含むものが挙げられる。 In the formula (I), the organic group in the group represented by the formula: -O- (organic group), -NH- (organic group) or -S- (organic group) represented by R includes groups derived from active ingredients of pharmaceuticals, luminescent groups and/or light absorbing groups (these groups may be groups derived from active ingredients of diagnostic agents), or functional groups (for example, maleimide groups and alkyne groups useful as reaction reagents).

前記式（Ｉ）において、Ｒが塩素原子である化合物は、次式（II）：
ＨＯＣＨ_２ＣＦ_２Ｏ－（ＣＦ_２ＣＦ_２Ｏ）_ｎ－ＣＦ_２ＣＨ_２ＯＨ（II）
（式中、ｎは１～７の整数を表す。）
で示されるフッ素化ポリエチレングリコールを塩化ホスホリルと反応させることにより製造することができる。 In the above formula (I), the compound in which R is a chlorine atom is represented by the following formula (II):
_HOCH2CF2O- ₍ _CF2CF2O ) _n- _CF2CH2OH ₍ _II )
(In the formula, n represents an integer of 1 to 7.)
The compound can be prepared by reacting a fluorinated polyethylene glycol represented by the formula: with phosphoryl chloride.

前記の反応は、通常、ピリジン、トリエチルアミン、Ｎ，Ｎ－ジイソプロピルエチルアミン（ＤＩＰＡ）、１，８－ジアザシクロ［５．４．０］ウンデカ－７－エン（ＤＢＵ）等の塩基の存在下に行う。塩化ホスホリルの使用量は、化合物（II）に対して、通常０．９～２当量、好ましくは０．９～１．２当量である。 The above reaction is usually carried out in the presence of a base such as pyridine, triethylamine, N,N-diisopropylethylamine (DIPA), or 1,8-diazacyclo[5.4.0]undec-7-ene (DBU). The amount of phosphoryl chloride used is usually 0.9 to 2 equivalents, preferably 0.9 to 1.2 equivalents, relative to compound (II).

溶媒としては、本反応を阻害しないものであれば特に限定さないが、例えば、無水ジクロロメタン、無水テトラヒドロフラン、無水アセトニトリル、無水１，４－ジオキサン、無水ジメチルホルムアミドが好適に使用される。 There are no particular limitations on the solvent, so long as it does not inhibit the reaction, but for example, anhydrous dichloromethane, anhydrous tetrahydrofuran, anhydrous acetonitrile, anhydrous 1,4-dioxane, and anhydrous dimethylformamide are preferably used.

反応は、－７０℃で開始し、特にその温度を維持せずに、時間経過で反応溶液の温度を室温に戻しながら行う。 The reaction is started at -70°C and is carried out without maintaining that temperature, allowing the temperature of the reaction solution to return to room temperature over time.

なお、前記式（II）において、ｎが１又は２である化合物、すなわちフッ素化トリエチレングリコール及びフッ素化テトラエチレングリコール（1H,1H,11H,11H-dodecafluoro-3,6,9-trioxaundecane-1,11-diol）は、市販されているので、これらを用いることができる。 In addition, compounds in which n is 1 or 2 in the above formula (II), i.e., fluorinated triethylene glycol and fluorinated tetraethylene glycol (1H,1H,11H,11H-dodecafluoro-3,6,9-trioxaundecane-1,11-diol), are commercially available, and these can be used.

前記式（Ｉ）において、Ｒが水酸基である化合物は、前記式（Ｉ）において、Ｒが塩素原子である化合物を水と反応させることにより製造することができる。 The compound of formula (I) in which R is a hydroxyl group can be produced by reacting the compound of formula (I) in which R is a chlorine atom with water.

反応温度は、通常１５～４０℃、好ましくは室温であり、反応時間は通常４０分～２時間、好ましくは５０分～１．５時間である。 The reaction temperature is usually 15 to 40°C, preferably room temperature, and the reaction time is usually 40 minutes to 2 hours, preferably 50 minutes to 1.5 hours.

前記式（Ｉ）において、Ｒが塩素原子である化合物は、反応性に富み、例えば、水酸基、アミノ基及び／又はチオール基（メルカプト基）を有する化合物と反応するので、種々の官能基（例えばマレイミド基、アルキン基）、機能性基（例えば発光性基）、医薬の有効成分に由来する基、診断薬の有効成分に由来する基を導入することができる。 In the formula (I), compounds in which R is a chlorine atom are highly reactive and react with compounds having, for example, a hydroxyl group, an amino group, and/or a thiol group (mercapto group), making it possible to introduce various functional groups (e.g., maleimide group, alkyne group), functional groups (e.g., luminescent groups), groups derived from active ingredients of pharmaceuticals, and groups derived from active ingredients of diagnostic agents.

前記式（Ｉ）において、Ｒは式：－Ｏ－（有機基）、－ＮＨ－（有機基）又は－Ｓ－（有機基）で示される基を表す。前記有機基の例としては、Ｃ１～Ｃ１５のアルキレン基と、その末端に反応性官能基とを有する基が挙げられる。前記末端の反応性官能基の例には、マレイミド基、アルキン基及び－ＮＨ_２・ＨＣｌが含まれる。この例の有機基を含む式（Ｉ）の化合物は、種々の試薬と反応（例えば、クリック反応）させることで、機能性基（例えば発光性基）、医薬の有効成分に由来する基、診断薬の有効成分に由来する基を導入することができる。本態様において、前記式（Ｉ）で示される化合物と反応させる試薬の例には、アルキン基及びマレイミド基を有する試薬とクリック反応可能な試薬として市販されている反応試薬及び従来公知の方法で製造した反応試薬いずれも含まれる。 In the formula (I), R represents a group represented by the formula: -O- (organic group), -NH- (organic group) or -S- (organic group). Examples of the organic group include a group having a C1-C15 alkylene group and a reactive functional group at its terminal. Examples of the reactive functional group at the terminal include a maleimide group, an alkyne group and -NH ₂ ·HCl. The compound of formula (I) containing the organic group of this example can be reacted with various reagents (e.g., click reaction) to introduce a functional group (e.g., a luminescent group), a group derived from an active ingredient of a medicine, or a group derived from an active ingredient of a diagnostic agent. In this embodiment, examples of the reagent to be reacted with the compound represented by formula (I) include both commercially available reaction reagents capable of click reaction with reagents having an alkyne group and a maleimide group, and reaction reagents produced by conventionally known methods.

例えば、前記式（Ｉ）において、Ｒが塩素原子で、ｎが２である化合物は、以下のようにして、具体的には、ＯＨ基とマレイミド基又はアルキン基とを有する化合物とを反応させることで、－Ｃ_ｍＨ_２ｍ－マレイミド基、－Ｃ_ｍＨ_２ｍ－アルキン基をそれぞれ導入することができる。以下では、ｍ＝２の例を示したが、ｍについては特に制限されず、例えば、ｍは１～１６である。また、ｍが３以上の場合、２つのＣＨ_２の間に位置する１以上のＣＨ_２はＯ及び／又はＮＨで置換されていてもよく、すなわち、オキシアルキレン基（例えばＣ１～Ｃ１６のオキシアルキレン基であって、－（Ｃ_ｐＨ_２ｐＯ）_ｑ－；ｐは１～４、ｑは１～４）やアミノアルキレン基（例えばＣ１～Ｃ１６のアミノアルキレン基であって、－（Ｃ_ｐＨ_２ｐＮＨ）_ｑ－；ｐは１～４、ｑは１～４）であってもよい。 For example, in the above formula (I), a compound in which R is a chlorine atom and n is 2 can be reacted with a compound having an OH group and a maleimide group or an alkyne group as follows to introduce _{a -CmH2m} _- maleimide group or _{a -CmH2m} _- alkyne group, respectively. In the following, an example in which m=2 is shown, but m is not particularly limited, and m is, for example, 1 to 16. Furthermore, when m is 3 or more, one or more _CH2s located between two _CH2s may be substituted with O and/or NH, that is, an oxyalkylene group (for example, a C1 to C16 oxyalkylene group, -( _CpH2pO ) _q- ; _p is 1 to 4, q is 1 to 4) or an aminoalkylene group (for example, a C1 to C16 _{aminoalkylene group, -(CpH2pNH} ₎ _q- ; p is 1 to 4, q is 1 to 4).

前記のようにして、マレイミド基が導入された化合物は、チオール基を有する様々な試薬とクリック反応可能であり、アルキン基が導入された化合物は、アジド基（Ｎ_３－）を有する試薬とクリック反応可能であり、バリエーションに富んだ機能性の有機フッ素化合物を得ることができる。 As described above, a compound having a maleimide group introduced therein can undergo a click reaction with various reagents having a thiol group, and a compound having an alkyne group introduced therein can undergo a click reaction with a reagent having an azide group (N ₃ -), thereby making it possible to obtain organic fluorine compounds with a wide variety of functions.

前記のマレイミド基が導入された化合物とチオール基を有する試薬との反応の一例を以下に示す。 An example of the reaction between the compound having the maleimide group and a reagent having a thiol group is shown below.

（式中、ＤＢＵはジアザビシクロウンデセンを表し、ＤＭＦはジメチルホルムアミドを表す。）

(In the formula, DBU represents diazabicycloundecene, and DMF represents dimethylformamide.)

また、ＲがＣ１～Ｃ１５のアルキレン基と、その末端に－ＮＨ_２・ＨＣｌを有する化合物は、例えば、後述する実施例４及び実施例７の方法に準じて種々合成することができる。末端に－ＮＨ_２・ＨＣｌを有する化合物は、ハロゲン（例えば－Ｃｌ）を有する試薬と反応させることで、種々の官能基をＰＩＰＰＥＲに結合させることができる。 In addition, various compounds in which R is a C1-C15 alkylene group and has _-NH2.HCl at its terminal can be synthesized, for example, according to the methods of Examples 4 and 7 described below. Various functional groups can be bonded to PIPPER by reacting the compound having _-NH2.HCl at its terminal with a reagent having a halogen (e.g., -Cl).

前記式（Ｉ）で示される化合物は、水溶性であり、凝集などなく水中で安定に存在（存在の状態には、溶解状態及び分散状態のいずれも含まれる）するので、当該化合物と、水系溶媒とを含む組成物は、薬剤キャリア、^１９ＦＮＭＲ造影剤などの診断薬、生体マーカー、接着剤、光電子デバイス用材料、２Ｄ材料として有用である。前記水系溶媒としては、例えば水、水と混和可能な有機溶媒（エタノール、ジメチルスルホキシド、ジメチルホルムアミド、メタノール、アセトニトリル、テトラヒドロフランなど）と水の混合液が挙げられる。 The compound represented by formula (I) is water-soluble and exists stably in water without aggregation (the state of existence includes both a dissolved state and a dispersed state), so that a composition containing the compound and an aqueous solvent is useful as a drug carrier, a diagnostic agent such as a ¹⁹ F NMR contrast agent, a biomarker, an adhesive, a material for optoelectronic devices, and a 2D material. Examples of the aqueous solvent include water and a mixture of water and an organic solvent miscible with water (ethanol, dimethyl sulfoxide, dimethylformamide, methanol, acetonitrile, tetrahydrofuran, etc.).

また、前記式（Ｉ）で示される化合物と水系溶媒とを含む組成物は、更に無機イオンを含んでいてもよい。前記式（Ｉ）で示される化合物は、非共有電子対を有する酸素原子を環構成原子とするクラウンエーテル類似の環状構造を有する。この特徴から、環の内側にカチオンを包接可能である。包接されるカチオンは特に制限されない。例えば、前記式（Ｉ）においてｎ＝２の化合物では、アルカリ金属のカチオンを包接可能である。 The composition containing the compound represented by formula (I) and the aqueous solvent may further contain an inorganic ion. The compound represented by formula (I) has a cyclic structure similar to crown ether, in which an oxygen atom having an unshared electron pair is a ring-constituting atom. Due to this characteristic, it is possible to include a cation inside the ring. The cation to be included is not particularly limited. For example, in the compound of formula (I) where n=2, it is possible to include an alkali metal cation.

更に、前記式（Ｉ）で示される化合物は、アルカリ金属イオン、アルカリ土類金属イオン等の無機イオンを取り込む（包接する）能力を有し、無機イオン包接剤として有用である。例えば、無機化合物のＫＭｎＯ_４はイオン性化合物のため有機溶媒に不溶であるが、前記式（Ｉ）で示される化合物が存在すると、カリウムイオン（Ｋ^＋）が当該化合物に捕捉され、ベンゼンをはじめとする有機溶媒に溶けるようになる。したがって、ＫＦ、ＫＣＮ、ＮａＮ_３などに代表される難溶性アルカリ金属塩を有機溶媒中で効果的に用いることができ、有機合成上有用である。 Furthermore, the compound represented by the formula (I) has the ability to capture (enclose) inorganic ions such as alkali metal ions and alkaline earth metal ions, and is useful as an inorganic ion inclusion agent. For example, the inorganic compound _KMnO4 is an ionic compound and is insoluble in organic solvents, but when the compound represented by the formula (I) is present, potassium ions (K ⁺ ) are captured by the compound, and it becomes soluble in organic solvents such as benzene. Therefore, poorly soluble alkali metal salts such as KF, KCN, _NaN3 , etc. can be effectively used in organic solvents, which is useful in organic synthesis.

以下、実施例により本発明を更に具体的に説明するが、本発明の範囲は下記の実施例に限定されることはない。 The present invention will be explained in more detail below with reference to examples, but the scope of the present invention is not limited to the following examples.

（実施例１）ＰＩＰＰＥＲ－Ｃｌの合成

(Example 1) Synthesis of PIPPER-Cl

オーブンで乾燥させた１Ｌの三口丸底フラスコに、フッ素化テトラエチレングリコール（1H,1H,11H,11H-dodecafluoro-3,6,9-trioxaundecane-1,11-diol）（４．５２ｇ，１１．０ｍｍｏｌ）を無水ジクロロメタン３００ｍＬに溶解し、温度を－７０℃に低下させ、白色懸濁液を得た。無水ジクロロメタン２００ｍＬ中の塩化ホスホリル（１．６５ｇ，１．０ｍＬ，１０．８ｍｍｏｌ）の溶液を、滴下漏斗を用いて滴下し、－７０℃を維持しつつ、１時間攪拌した後、ゆっくりと温度を室温に戻した。更に、６時間攪拌後、無水ピリジン（１．８７ｇ，２３．７ｍｍｏｌ）を注射器で滴下した。反応混合物を室温で更に２４時間攪拌した。溶液を窒素気流下溶媒留去し、目的とするＰＩＰＰＥＲ－Ｃｌを得た。ＭＡＬＤＩ－ＴＯＦＭＳは、マトリックスとしてtrans-2-[3-(4-tert-butylphenyl)-2-methyl-2-propenylidene]malononitrileを用いたネガティブイオンモードで、４７１．１４６Ｄａに、ＰＩＰＰＥＲ（ＰＩＰＰＥＲ－ＯＨ）の単一ピークを示し、反応性のＰＩＰＰＥＲ－Ｃｌ中間体の形成を示した。生成物を CH₂Cl₂/Hexane (1:1) で精製するため、－２０℃でＮ_２の下で再結晶した。
¹H NMR (CDCl₃, 298 K, 600 MHz): δ = 4.39 (m, -CH₂ CF₂), 4.64 (m, -CH₂ CF₂) ppm.¹³C NMR (CDCl₃, 298 K, 150 MHz): δ = 66.07 (m, -CH₂), 114.22 (m, -CF₂), 114.50 (m, -CF₂), 120.21 (td, J_C,F= 274.4 Hz, 10.6 Hz -CF₂) ppm.
¹⁹F NMR (CDCl₃, 298 K, 565 MHz): δ = -77.93 (m), -78.31 (m), -89.24 (m), -89.60 (s), -90.09 (m) ppm.
³¹P NMR (CDCl₃, 298 K, 243 MHz): δ = 5.24.
MALDI-TOF MS: for C₈H₄F₁₂O₇P [M-H]^-, calculated: 471.07 Da, found: 471.15 Da In an oven-dried 1 L three-necked round-bottom flask, fluorinated tetraethylene glycol (1H,1H,11H,11H-dodecafluoro-3,6,9-trioxaundecane-1,11-diol) (4.52 g, 11.0 mmol) was dissolved in 300 mL of anhydrous dichloromethane, and the temperature was lowered to -70°C to obtain a white suspension. A solution of phosphoryl chloride (1.65 g, 1.0 mL, 10.8 mmol) in 200 mL of anhydrous dichloromethane was added dropwise using a dropping funnel, and the mixture was stirred for 1 hour while maintaining the temperature at -70°C, and then the temperature was slowly returned to room temperature. After stirring for an additional 6 hours, anhydrous pyridine (1.87 g, 23.7 mmol) was added dropwise using a syringe. The reaction mixture was stirred at room temperature for an additional 24 hours. The solvent was distilled off from the solution under a nitrogen stream to obtain the desired PIPPER-Cl. MALDI-TOF MS in the negative ion mode using trans-2-[3-(4-tert-butylphenyl)-2-methyl-2-propenylidene]malononitrile as the matrix showed a single peak for PIPPER (PIPPER-OH) at 471.146 Da, indicating the formation of a reactive PIPPER-Cl intermediate. The product was purified with CH ₂ Cl ₂ /Hexane (1:1) and recrystallized under N ₂ at −20 °C.
^1H NMR ( _CDCl3 , 298 K, 600 MHz): δ = 4.39 (m _, _-CH2CF2 ), 4.64 (m, _-CH2CF2 ) ppm. ^13C NMR ( _CDCl3 , 298 K, 150 MHz): δ = 66.07 (m, _-CH2 ) _, 114.22 (m, _-CF2 ), 114.50 (m, _-CF2 ), 120.21 (td, _JC,F = 274.4 Hz, 10.6 Hz _-CF2 ) ppm.
^19F NMR ( _CDCl3 , 298 K, 565 MHz): δ = -77.93 (m), -78.31 (m), -89.24 (m), -89.60 (s), -90.09 (m) ppm.
^31P NMR ( _CDCl3 , 298 K, 243 MHz): δ = 5.24.
MALDI _- TOF MS: for _C8H4F12O7P [MH] ^- , calculated: _471.07 Da _, found: 471.15 Da

（実施例２）ＰＩＰＰＥＲ（ＰＩＰＰＥＲ－ＯＨ）の合成

(Example 2) Synthesis of PIPPER (PIPPER-OH)

実施例１で得られた粗ＰＩＰＰＥＲ－Ｃｌを１００ｍＬの丸底フラスコに取り、Milli-Q^TM 水３０ｍＬを加えた。混合物を１時間攪拌した後、蒸発乾固させた。白色粘着性の粗生成物を、１０％酢酸含有酢酸エチルを用いた、シリカゲル上のカラムクロマトグラフィーで精製し、目的とするＰＩＰＰＥＲ（ＰＩＰＰＥＲ－ＯＨ）を無色粘性の液状物として得た（収量２．１０ｇ，４．４４ｍｍｏｌ，収率４１％）。
¹H NMR (D₂O, 298 K, 600 MHz): δ = 4.34 (q, ³J_H,F = 7.2 Hz, 4H, -CH₂) ppm.
¹³C NMR (D₂O, 298 K, 150 MHz): δ = 64.24 (t, J_C,F = 36.8 Hz, -CH₂), 113.93 (m, -CF₂), 114.24 (m, -CF₂), 121.51 (td, J_C,F = 274.5 Hz, 10.4 Hz, -CF₂) ppm.
¹⁹F NMR (D₂O, 298 K, 565 MHz): δ = -78.6, -89.5, -89.6 ppm.
³¹P NMR (D₂O, 298 K, 243 MHz): δ = -2.28.
MALDI-TOF MS: for C₈H₄F₁₂O₇P [M-H]^-, calculated: 471.07 Da, found: 471.15 Da The crude PIPPER-Cl obtained in Example 1 was placed in a 100 mL round-bottom flask and 30 mL of Milli-Q ^™ water was added. The mixture was stirred for 1 hour and then evaporated to dryness. The white sticky crude product was purified by column chromatography on silica gel using 10% acetic acid in ethyl acetate to obtain the desired PIPPER (PIPPER-OH) as a colorless viscous liquid (yield 2.10 g, 4.44 mmol, 41%).
^1H NMR ( _D2O , 298K, 600MHz): δ = 4.34 (q, ^3JH _,F = 7.2Hz, 4H, _-CH2 ) ppm.
^13C NMR ( _D2O , 298 K, 150 MHz): δ = 64.24 (t, JC _,F = 36.8 Hz, _-CH2 ), 113.93 (m, _-CF2 ), 114.24 (m, _-CF2 ), 121.51 (td, _JC,F = 274.5 Hz, 10.4 Hz, _-CF2 ) ppm.
^19F NMR ( _D2O , 298 K, 565 MHz): δ = -78.6, -89.5, -89.6 ppm.
^31P NMR ( _D2O , 298K, 243MHz): δ = -2.28.
MALDI _- TOF MS: for _C8H4F12O7P [MH] ^- , calculated: _471.07 Da _, found: 471.15 Da

（実施例３）ＰＩＰＰＥＲ－ＭＡＬの合成

(Example 3) Synthesis of PIPPER-MAL

実施例１と同様にして得られた粗ＰＩＰＰＥＲ－Ｃｌを無水トルエン２００ｍＬで希釈した。次いで、ピリジン１．０ｍＬ及びＮ－（２－ヒドロキシエチル）マレイミド（１．５０ｇ，１０．６ｍｍｏｌ）を窒素気流下加えた。混合物を１００℃で１時間加熱した後、薄層クロマトグラフィーに付した。溶媒を留去した後、残渣をジクロロメタン１００ｍＬで抽出し、Milli-Q^TM 水１５０ｍＬで洗浄した。有機層を無水硫酸ナトリウム上で乾燥し、溶媒を除去した。粗生成物を、０～２０％酢酸エチル含有ジクロロメタンを用いた、シリカゲル上のカラムクロマトグラフィーで精製し、目的とするＰＩＰＰＥＲ－ＭＡＬを白色固体として得た（収量２．４ｇ，４．０３ｍｍｏｌ，収率３７％）。
¹H NMR (CD₂Cl₂, 298 K, 600 MHz): δ = 3.83 (t, ³J = 5.4 Hz, 2H, -NHCH₂), 4.26 (m, 2H, -OCH₂), 4.31 (m, 2H, -OCF₂CH₂O), 4.48 (m, 2H, -OCF₂CH₂O), 6.75 (s, 2H, -CH) ppm.
¹³C NMR (CD₂Cl₂, 298 K, 150 MHz): δ = 37.86 (q, ³J_C,F = 6.0 Hz, -CH₂), 65.61 (m, -CH₂), 66.01 (q, ³J_C,F = 6.0 Hz, -CH₂), 114.17 (m, -CF₂), 114.50 (m, -CF₂), 120.81 (td, ³J_C,F = 276, 9.0 Hz, -CF₂), 134.41 (s, -CH), 170.59 (s, -C=O) ppm.
¹⁹F NMR (CD₂Cl₂, 298 K, 565 MHz): δ = -78.18 (ddt, J_C,F/F,F = 497.2, 146.9, 17.0, 11.3 Hz), -79.06 (ddt, ³J_C,F/F,F = 146.9, 17.0, 5.7 Hz), -88.83 (ddt, J_C,F/F,F = 146.9, 17.0, 5.7 Hz), -89.74 (t, J_C,F/F,F = 5.7 Hz), -90.60 (ddt, J_C,F/F,F = 146.9, 17.0, 5.7 Hz) ppm.
³¹P NMR (CD₂Cl₂, 298 K, 243 MHz): δ = -2.52 ppm.
MALDI-TOF MS: for C₁₄H₁₀F₁₂NO₉P・Na [M+Na]⁺, calculated: 617.98 Da, found: 618.40 Da Crude PIPPER-Cl obtained as in Example 1 was diluted with 200 mL of anhydrous toluene. Then, 1.0 mL of pyridine and N-(2-hydroxyethyl)maleimide (1.50 g, 10.6 mmol) were added under a nitrogen stream. The mixture was heated at 100° C. for 1 hour and then subjected to thin layer chromatography. After distilling off the solvent, the residue was extracted with 100 mL of dichloromethane and washed with 150 mL of Milli-Q ^™ water. The organic layer was dried over anhydrous sodium sulfate and the solvent was removed. The crude product was purified by column chromatography on silica gel using 0-20% ethyl acetate in dichloromethane to obtain the desired PIPPER-MAL as a white solid (yield 2.4 g, 4.03 mmol, 37%).
^1H NMR ( _CD2Cl2 , ₂₉₈ K, 600 MHz): δ = 3.83 (t, ^3J = 5.4 Hz, 2H, _-NHCH2 ), 4.26 (m, 2H, _-OCH2 ) _, 4.31 (m, _2H , _-OCF2CH2O ), 4.48 (m, 2H _, -OCF2CH2O), 6.75 (s, 2H, -CH) ppm.
^13C NMR ( _CD2Cl2 , ₂₉₈ K, 150 MHz): δ = 37.86 (q, ^3JC _,F = 6.0 Hz, _-CH2 ), 65.61 (m, _-CH2 ), 66.01 (q, ^3JC _,F = 6.0 Hz, _-CH2 ), 114.17 (m, _-CF2 ), 114.50 (m, _-CF2 ), 120.81 (td, ^3JC _,F = 276, 9.0 Hz, _-CF2 ), 134.41 (s, -CH), 170.59 (s, -C=O) ppm.
^19F NMR ( _CD2Cl2 , ₂₉₈ K, 565 MHz): δ = -78.18 (ddt, JC _,F/F,F = 497.2, 146.9, 17.0, 11.3 Hz), -79.06 (ddt, ³ JC _,F/F,F = 146.9, 17.0, 5.7 Hz), -88.83 (ddt, JC _,F/F,F = 146.9, 17.0, 5.7 Hz), -89.74 (t, _JC,F/F,F = 5.7 Hz), -90.60 (ddt, _JC,F/F,F = 146.9, 17.0, 5.7 Hz) ppm.
^31P NMR ( _CD2Cl2 , _298K , 243MHz): δ = -2.52 ppm.
MALDI-TOF MS: for _{C14H10F12NO9P}・Na [ _M ₊ Na] ⁺ , calculated: 617.98 Da, _found : 618.40 Da

（実施例４）ＰＩＰＰＥＲ－ＮＨ_２・ＨＣｌの合成

(Example 4) Synthesis of PIPPER- _NH2.HCl

実施例１と同様にして得られた粗ＰＩＰＰＥＲ－Ｃｌを無水トルエン２００ｍＬで希釈した。次いで、ピリジン１．０ｍＬ及びＮ－（tert－ブトキシカルボニル）エタノールアミン（１．７４ｇ，１０．８ｍｍｏｌ）を窒素気流下加えた。混合物を１００℃で１時間加熱した後、薄層クロマトグラフィーに付した。溶媒を留去した後、粗生成物を、０～１０％酢酸エチル含有ジクロロメタンを用いた、シリカゲル上のカラムクロマトグラフィーで精製し、Ｂｏｃ－ＮＨ体を白色固体として得た（収量２．５２ｇ，４．１０ｍｍｏｌ，収率３８％）。Ｂｏｃ－ＮＨ体を１００ｍＬの丸底フラスコに取り、酢酸エチル３０ｍＬに溶解した。次いで、エタノール３０ｍＬ及び塩化アセチル３．０ｍＬ（３．３０ｇ，４２．０ｍｍｏｌ）を加え、混合物を２４時間室温で撹拌した。溶媒を留去し、更に精製することなく、核磁気共鳴分光法及びＭＡＬＤＩ－ＴＯＦＭＳで特徴づけられる目的化合物ＰＩＰＰＥＲ－ＮＨ_２・ＨＣｌを得た。
¹H NMR (CD₃OD, 298 K, 600 MHz): δ = 3.36 (t, ³J = 5.4 Hz, 2H, -NHCH₂), 4.48 (dd, ³J = 9.4, 5.4 Hz, 2H, -OCH₂), 4.70 (m, 2H, -OCF₂CH₂O), 4.75 (m, 2H, -OCF₂CH₂O) ppm.
¹³C NMR (CD₃OD, 298 K, 150 MHz): δ = 41.10 (m, -CH₂), 66.76 (m, -CH₂), 67.21 (m, -CH₂), 115.82 (m, -CF₂), 116.12 (m, -CF₂), 122.71 (td, ³J_C,F = 276, 9.2 Hz, -CF₂) ppm.
¹⁹F NMR (CD₃OD, 298 K, 565 MHz): δ = -79.34 (m), -80.02 (m), -90.14 (m), -90.91 (s), -91.44 (m) ppm.
³¹P NMR (CD₃OD, 298 K, 243 MHz): δ = -3.14 ppm.
MALDI-TOF MS: for C₁₀H₁₀F₁₂NO₇P・Na [M+Na]⁺, calculated: 538.13 Da, found: 538.965 Da (PIPPER-NH₂・HCl), 471.254 Da (PIPPER-OH). The crude PIPPER-Cl obtained as in Example 1 was diluted with 200 mL of anhydrous toluene. Then, 1.0 mL of pyridine and N-(tert-butoxycarbonyl)ethanolamine (1.74 g, 10.8 mmol) were added under a nitrogen stream. The mixture was heated at 100° C. for 1 h and then subjected to thin layer chromatography. After distilling off the solvent, the crude product was purified by column chromatography on silica gel using 0-10% ethyl acetate in dichloromethane to give the Boc-NH compound as a white solid (yield 2.52 g, 4.10 mmol, 38%). The Boc-NH compound was taken up in a 100 mL round bottom flask and dissolved in 30 mL of ethyl acetate. Then, 30 mL of ethanol and 3.0 mL of acetyl chloride (3.30 g, 42.0 mmol) were added and the mixture was stirred at room temperature for 24 h. The solvent was evaporated to give the target compound PIPPER-NH ₂ ·HCl, which was characterized by nuclear magnetic resonance spectroscopy and MALDI-TOF MS without further purification.
^1H NMR ( _CD3OD , 298K, 600MHz): δ = 3.36 (t, ^3J = 5.4Hz, 2H, _-NHCH2 ) _, 4.48 (dd, ^3J = 9.4, 5.4Hz _, 2H, _-OCH2 ), 4.70 (m, 2H _, -OCF2CH2O), 4.75 (m, 2H, _-OCF2CH2O ) ppm.
^13C NMR (CD3OD, _298K , 150MHz): δ = 41.10 (m, _-CH2 ), 66.76 (m, _-CH2 ), 67.21 (m, _-CH2 ) _, 115.82 (m, -CF2), 116.12 (m, _-CF2 ), 122.71 (td, ^3JC _,F = 276, 9.2Hz, _-CF2 ) ppm.
^19F NMR ( _CD3OD , 298K, 565MHz): δ = -79.34 (m), -80.02 (m), -90.14 (m), -90.91 (s), -91.44 (m) ppm.
^31P NMR ( _CD3OD , 298K, 243MHz): δ = -3.14ppm.
MALDI-TOF MS: for _{C10H10F12NO7P}・Na [M ₊ Na] ⁺ , calculated: 538.13 Da, found: _538.965 Da (PIPPER- _NH2・HCl), 471.254 Da (PIPPER-OH ₎ .

（実施例５）ＰＩＰＰＥＲ－Guanidinium Chlorideの合成

(Example 5) Synthesis of PIPPER-Guanidinium Chloride

実施例１と同様にして得られた粗ＰＩＰＰＥＲ－Ｃｌを無水トルエン２００ｍＬで希釈した。次いで、ピリジン１．０ｍＬ及びCarbamic acid, N,N'-[[2-(2-hydroxyethoxy)ethyl]carbonimidoyl]bis-,C,C'-bis(2,2-dimethylethyl)ester（３．７５ｇ，１０．８ｍｍｏｌ）を窒素気流下加えた。混合物を１００℃で１時間加熱した後、薄層クロマトグラフィーに付した。溶媒を留去した後、粗生成物を、３０％酢酸エチル含有ヘキサンを用いた、シリカゲル上のカラムクロマトグラフィーで精製し、Ｂｏｃ－Guanidinium体を白色固体として得た（収量２．３０ｇ，２．８７ｍｍｏｌ，収率２７％）。Ｂｏｃ－Guanidinium体を１００ｍＬの丸底フラスコに取り、酢酸エチル１５ｍＬに溶解した。次いで、エタノール１５ｍＬ及び塩化アセチル６．０ｍＬ（６．６０ｇ，８４．０ｍｍｏｌ）を加え、混合物を２４時間室温で撹拌した。溶媒を留去し、更に精製することなく、核磁気共鳴分光法及びＭＡＬＤＩ－ＴＯＦＭＳで特徴づけられる目的化合物ＰＩＰＰＥＲ－Guanidinium Chlorideを得た。
¹H NMR (CD₃OD, 298 K, 600 MHz): δ = 3.40 (d, ³J = 5.4 Hz, 2H, -NHCH₂), 3.68 (br, 2H, -OCH₂), 3.77 (br, 2H, -OCH₂), 4.34 (t, ³J = 5.4 Hz, 2H, -OCH₂), 4.59 (m, 2H, -OCF₂CH₂O), 4.66 (m, 2H, -OCF₂CH₂O) ppm.
¹³C NMR (CD₃OD, 298 K, 150 MHz): δ = 42.71 (m, -CH₂), 66.73(m, -CH₂), 70.02 (m, -CH₂), 70.35 (m, -CH₂), 70.86 (m, -CF₂), 115.48 (m, -CF₂), 115.80 (m, -CF₂), 122.48 (td, ³J_C,F = 276, 9.0 Hz, -CF₂), 159.08 (s, -N₂C=NH) ppm.
¹⁹F NMR (CD₃OD, 298 K, 565 MHz): δ = -79.28 (m), -79.90 (m), -90.09 (m), -90.84 (s), -91.32 (m) ppm.
³¹P NMR (CD₃OD, 298 K, 243 MHz): δ = -2.87 ppm.
MALDI-TOF MS: for C₁₃H₁₇F₁₂N₃O₈P [M-Cl]⁺, calculated: 602.06 Da, found: 602.24 Da (PIPPER-Guanidinium⁺). The crude PIPPER-Cl obtained in the same manner as in Example 1 was diluted with 200 mL of anhydrous toluene. Then, 1.0 mL of pyridine and carbamic acid, N,N'-[[2-(2-hydroxyethoxy)ethyl]carbonimidoyl]bis-,C,C'-bis(2,2-dimethylethyl)ester (3.75 g, 10.8 mmol) were added under a nitrogen stream. The mixture was heated at 100°C for 1 hour and then subjected to thin layer chromatography. After distilling off the solvent, the crude product was purified by column chromatography on silica gel using 30% ethyl acetate in hexane to obtain the Boc-Guanidinium compound as a white solid (yield 2.30 g, 2.87 mmol, 27%). The Boc-Guanidinium compound was placed in a 100 mL round-bottom flask and dissolved in 15 mL of ethyl acetate. Then, 15 mL of ethanol and 6.0 mL of acetyl chloride (6.60 g, 84.0 mmol) were added, and the mixture was stirred at room temperature for 24 hours. The solvent was distilled off to obtain the target compound PIPPER-Guanidinium Chloride without further purification, which was characterized by nuclear magnetic resonance spectroscopy and MALDI-TOF MS.
^1H NMR ( _CD3OD , 298K, 600MHz): δ = 3.40 (d, ^3J = 5.4Hz, 2H, _-NHCH2 ), 3.68 (br, 2H, _-OCH2 ), 3.77 (br, 2H, _-OCH2 ), 4.34 (t, ^3J = 5.4Hz, 2H, _-OCH2 ) _, 4.59 (m, 2H, -OCF2CH2O), 4.66 (m, 2H _, _-OCF2CH2O ) ppm _.
^13C NMR (CD3OD, _298K , 150MHz): δ = 42.71 (m, _-CH2 ), 66.73(m, _-CH2 ), 70.02 (m, _-CH2 ), 70.35 (m, _-CH2 ), 70.86 (m, _-CF2 ), 115.48 (m, _-CF2 ), 115.80 (m, _-CF2 ), 122.48 (td, ^3JC _,F = 276, 9.0Hz, _-CF2 ), 159.08 (s, _-N2C =NH) ppm.
^19F NMR (CD3OD, _298K , 565MHz): δ = -79.28 (m), -79.90 (m), -90.09 (m), -90.84 (s), -91.32 (m) ppm.
^31P NMR ( _CD3OD , 298K, 243MHz): δ = -2.87ppm.
MALDI-TOF MS: for _{C13H17F12N3O8P} [ _M -Cl] ₊ ^, calculated: 602.06 Da, found: _602.24 Da (PIPPER-Guanidinium ⁺ ₎ .

（実施例６）ＰＩＰＰＥＲ－Alkyneの合成

(Example 6) Synthesis of PIPPER-Alkyne

実施例３において、Ｎ－（２－ヒドロキシエチル）マレイミドの代わりに３－ブチン－１－オールを用いることで、ＰＩＰＰＥＲ－Alkyneを得た。
¹H NMR (CDCl₃, 298 K, 600 MHz): δ = 2.06 (t, 4J = 2.6 Hz, 1H, alkyne-H), 2.62 (td, 3J = 6.6, 4J 2.6 Hz, 2H, alkyne-CH₂), 4.21 (m, 2H, -OCH₂), 4.31 (m, 2H, -OCF₂CH₂O), 4.50 (m, 2H, -OCF₂CH₂O) ppm.
¹³C NMR (CDCl₃, 298 K, 600 MHz): δ = 20.75 (m, -CH₂), 65.60 (m, -CH₂), 66.80 (m, -CH₂), 71.02 (d, J_C,F/P= 40.5 Hz, -CH), 78.80 (d, J_C,F/P= 13.5 Hz, -CH), 114.02 (m, -CF₂), 114.54 (m, -CF₂), 120.64 (td, 3J_C,F= 276, 9.0 Hz, -CF₂)ppm.
¹⁹F NMR (CDCl₃, 298 K, 565 MHz): δ = -77.71 (m), -78.79 (m), -88.37 (m), -89.47 (s), -90.52 (m) ppm.
³¹P NMR (CDCl₃, 298 K, 243 MHz): δ = -3.22 ppm.
MALDI-TOF MS: for C₁₂H₉F₁₂O₇P・K [M+K]⁺, calculated: 562.95 Da, found: 563.011 Da (PIPPER-Alkyne + K⁺). In Example 3, 3-butyn-1-ol was used in place of N-(2-hydroxyethyl)maleimide to obtain PIPPER-Alkyne.
^1H NMR ( _CDCl3 , 298 K, 600 MHz): δ = 2.06 (t, 4J = 2.6 Hz, 1H, alkyne-H), 2.62 (td, 3J = 6.6, 4J 2.6 Hz, 2H, alkyne- _CH2 ), 4.21 (m, 2H, _-OCH2 ) _, 4.31 (m, 2H _, -OCF2CH2O), 4.50 (m, 2H _, _-OCF2CH2O ) ppm.
^13C NMR ( _CDCl3 , 298 K, 600 MHz): δ = 20.75 (m, _-CH2 ), 65.60 (m, _-CH2 ), 66.80 (m, _-CH2 ), 71.02 (d, _JC,F/P = 40.5 Hz, -CH), 78.80 (d, _JC,F/P = 13.5 Hz, -CH), 114.02 (m, _-CF2 ), 114.54 (m, _-CF2 ), 120.64 (td, 3JC _,F = 276, 9.0 Hz, _-CF2 ) ppm.
^19F NMR ( _CDCl3 , 298 K, 565 MHz): δ = -77.71 (m), -78.79 (m), -88.37 (m), -89.47 (s), -90.52 (m) ppm.
^31P NMR ( _CDCl3 , 298 K, 243 MHz): δ = -3.22 ppm.
MALDI-TOF MS: for _C12H9F12O7PK [M+K] ₊ ^, calculated: 562.95 Da _, found: 563.011 Da (PIPPER-Alkyne + K ⁺ ₎ .

精製したＰＩＰＰＥＲ－Alkyneを特別な操作を行わずにＭＡＬＤＩ－ＴＯＦＭＳにかけたところ、Ｋ^＋を補足したＰＩＰＰＥＲ－Alkyneのピークを観測し、以下に示すカリウムイオン包接体（Cal. m/z=562.95）が形成されたことを確認した。Ｋ^＋はＭＡＬＤＩ－ＴＯＦＭＳを行う際に用いるマトリックスに由来するものと考えられる。 When the purified PIPPER-Alkyne was subjected to MALDI-TOF MS without any special operations, the peak of PIPPER-Alkyne with K ⁺ was observed, and it was confirmed that the potassium ion inclusion complex shown below (Cal. m/z = 562.95) was formed. K ⁺ is considered to be derived from the matrix used when performing MALDI-TOF MS.

（実施例７）ＰＩＰＰＥＲ－ＮＨ－ＮＨＢＯＣの合成

(Example 7) Synthesis of PIPPER-NH-NHBOC

実施例４において、Ｎ－（tert－ブトキシカルボニル）エタノールアミンの代わりにＮ－（tert－ブトキシカルボニル）トリメチレンジアミンを用いることで、ＰＩＰＰＥＲ－ＮＨ－ＮＨＢＯＣを得た。
¹H NMR (CD₂Cl₂, 298 K, 600 MHz): δ = 1.42 (s, 9H, -Boc), 1.60 (m, 2H, - CH₂), 2.96 (m, 2H, -NHCH₂), 3.20 (m, 2H, -NHCH₂), 4.05 (br, 1H, -NH), 4.25 (m, 2H, -OCF₂CH₂O), 4.44 (m, 2H, -OCF₂CH₂O), 4.76 (br, 1H, -NH) ppm.
¹³C NMR (CD₂Cl₂, 298 K, 150 MHz): δ = 28.08 (s, -CH₂), 31.75 (d, 3J_C,F= 4.5 Hz, -CH₂), 36.71 (s, -CH₂), 38.04 (s, -CH₂), 64.28 (m, -CH₂), 79.44 (s, -OCMe₃), 114.18 (m, -CF₂), 115.58 (m, -CF₂), 121.30 (td, ³J_C,F= 276, 9.0 Hz, -CF₂), 156.88 (s, O=C-NH) ppm.
¹⁹F NMR (CD₂Cl₂, 298 K, 565 MHz): δ = -77.54 (m), -79.22 (m), -88.20 (m), -89.70 (s), -91.17 (m)ppm.
³¹P NMR (CD₂Cl₂, 298 K, 243 MHz): δ = -8.70ppm.
MALDI-TOF MS: for C₁₁H₁₄F₁₂N₂O₆P⁺[M+H]⁺, calculated: 529.04 Da, found: 529.515 Da (PIPER-NH-NH₃ ⁺). In Example 4, N-(tert-butoxycarbonyl)trimethylenediamine was used in place of N-(tert-butoxycarbonyl)ethanolamine to obtain PIPPER-NH-NHBOC.
^1H NMR ( _CD2Cl2 , ₂₉₈ K, 600 MHz): δ = 1.42 (s, 9H, -Boc), 1.60 (m, 2H, _-CH2 ), 2.96 (m, 2H, _-NHCH2 ), 3.20 (m, 2H, _-NHCH2 ), 4.05 (br, 1H, -NH) _, 4.25 (m, 2H _, -OCF2CH2O), 4.44 (m, 2H _, -OCF2CH2O), _4.76 (br, 1H, -NH) ppm.
^13C NMR ( _CD2Cl2 , ₂₉₈ K, 150 MHz): δ = 28.08 (s, _-CH2 ), 31.75 (d, 3J _C,F = 4.5 Hz, _-CH2 ), 36.71 (s, -CH2) _, 38.04 (s, _-CH2 ), 64.28 (m, _-CH2 ), 79.44 (s, _-OCMe3 ), 114.18 (m, _-CF2 ), 115.58 (m, _-CF2 ), 121.30 (td, ^3J _C,F = 276, 9.0 Hz, _-CF2 ), 156.88 (s, O=C-NH) ppm.
^19F NMR ( _CD2Cl2 , ₂₉₈ K, 565 MHz): δ = -77.54 (m), -79.22 (m), -88.20 (m), -89.70 (s), -91.17 (m)ppm.
^31P NMR ( _CD2Cl2 , _298K , 243MHz): δ = -8.70ppm.
MALDI-TOF MS: for _{C11H14F12N2O6P} ₊ [ _M +H] ₊ _, calculated: 529.04 Da, found ^: 529.515 Da (PIPER-NH- _NH3 ⁺ ⁾ .

（実施例８）ＰＩＰＰＥＲ－ＮＨ－ＮＢＤの合成

(Example 8) Synthesis of PIPPER-NH-NBD

（１）ＰＩＰＰＥＲ－ＮＨ－ＮＨ_３Ｃｌの合成
実施例４と同様に脱保護させて、ＰＩＰＰＥＲ－ＮＨ－ＮＨ_３Ｃｌを得た。
（２）ＰＩＰＰＥＲ－ＮＨ－ＮＢＤの合成
ＰＩＰＰＥＲ－ＮＨ－ＮＨ_３Ｃｌ及び４－クロロ－７－ニトロベンゾフラザンをトリエチルアミン１当量の存在下、アセトニトリル中、室温で１２時間反応させて、ＰＩＰＰＥＲ－ＮＨ－ＮＢＤを得た（収率７８％）。
¹H NMR (CDCl₃, 298 K, 600 MHz): δ = 1.97 (m, 2H,-CH₂), 3.17 (m, 2H, - CH₂), 3.21 (m, 1H, -NH), 3.62 (dd, ³J = 8.4, 5.4 Hz, 2H, ,-CH₂), 4.32 (m, 2H, -OCF₂CH₂O), 4.50 (m, 2H, -OCF₂CH₂O), 6.20 (d, ³J = 8.4 Hz, 1H, ,NBD-CH), 7.00 (t, ³J = 5.4 Hz 1H, -NH), 8.49 (d, ³J = 8.4 Hz, 1H, ,NBD-CH)ppm.
¹³C NMR (CD₂Cl₂, 298 K, 150 MHz): δ = 30.35 (s, -CH₂), 39.35 (d, ³J_C,F= 4.5 Hz, -CH₂), 41.68 (s, -CH₂), 65.08 (m, -CH₂), 99.82 (s, -Ar-C), 115.49 (m, -CF₂), 115.06 (m, -CF₂), 122.62 (td, ³J_C,F= 276, 9.0 Hz, -CF₂), 123.70 (s, Ar-C), 138.25 (s, Ar-C), 145.40 (s, -Ar-C), 145.78 (s, -Ar-C), 145.92 (s, -Ar-C) ppm.
¹⁹F NMR (CDCl₃, 298 K, 565 MHz): δ = -77.47 (m), -79.21 (m), -87.96 (m), -89.66 (s), -91.28 (m)ppm.
³¹P NMR (CD₂Cl₂, 298 K, 243 MHz): δ = 9.04 ppm.
MALDI-TOF MS: for C₁₇H₁₂F₁₂N₅O₉P[M-H]^-, calculated: 690.03 Da, found: 690.384 Da (PIPER-NH-NBD). (1) Synthesis of PIPPER-NH-NH ₃ Cl PIPPER-NH-NH ₃ Cl was obtained by deprotection in the same manner as in Example 4.
(2) Synthesis of PIPPER-NH-NBD PIPPER-NH-NH ₃ Cl and 4-chloro-7-nitrobenzofurazan were reacted in the presence of 1 equivalent of triethylamine in acetonitrile at room temperature for 12 hours to obtain PIPPER-NH-NBD (yield 78%).
^1H NMR ( _CDCl3 , 298 K, 600 MHz): δ = 1.97 (m, 2H, _-CH2 ), 3.17 (m, 2H, _-CH2 ), 3.21 (m, 1H,-NH), 3.62 (dd, ³ J = 8.4, 5.4 _Hz , 2H, , _-CH2 ) _, 4.32 (m, 2H, -OCF2CH2O), 4.50 (m, 2H, -OCF2CH2O) _, 6.20 (d, ³ J = _8.4 Hz, 1H, ,NBD-CH), 7.00 (t, ³ J = 5.4 Hz 1H, -NH), 8.49 (d, ³ J = 8.4 Hz, 1H, ,NBD-CH)ppm.
^13C NMR ( _CD2Cl2 , ₂₉₈ K, 150 MHz): δ = 30.35 (s, _-CH2 ), 39.35 (d, ^3JC _,F = 4.5 Hz, _-CH2 ), 41.68 (s, -CH2), 65.08 (m, _-CH2 ), 99.82 (s, -Ar-C) _, 115.49 (m, -CF2), 115.06 (m, _-CF2 ) _, 122.62 (td, ^3JC _,F = 276, 9.0 Hz, _-CF2 ), 123.70 (s, Ar-C), 138.25 (s, Ar-C), 145.40 (s, -Ar-C), 145.78 (s, -Ar-C), 145.92 (s, -Ar-C) ppm.
^19F NMR ( _CDCl3 , 298 K, 565 MHz): δ = -77.47 (m), -79.21 (m), -87.96 (m), -89.66 (s), -91.28 (m)ppm.
^31P NMR ( _CD2Cl2 , _298K , 243MHz): δ = 9.04ppm.
MALDI-TOF MS: for _{C17H12F12N5O9P} [MH _] ^- , calculated: _690.03 Da, _found : 690.384 Da (PIPER-NH _- NBD).

（実施例９）ＰＩＰＰＥＲ－Ｓ－Phenylethaneの合成

(Example 9) Synthesis of PIPPER-S-Phenylethane

実施例４において、Ｎ－（tert－ブトキシカルボニル）エタノールアミンの代わりに２－フェニルエタンチオールを用いる以外は同様にして、ＰＩＰＰＥＲ－Ｓ－Phenylethaneを得た。
¹H NMR (CDCl₃, 298 K, 600 MHz): δ = 1.42 (s, 9H, -Boc), 1.60 (m, 2H, - CH₂), 2.96 (m, 2H, -NHCH₂), 3.20 (m, 2H, -NHCH₂), 4.05 (br, 1H, -NH), 4.25 (m, 2H, -OCF₂CH₂O), 4.44 (m, 2H, -OCF₂CH₂O), 4.76 (br, 1H, -NH) ppm.
¹³C NMR (CDCl₃, 298 K, 150 MHz): δ = 29.85 (m, -CH₂), 40.41 (m, -CH₂), 66.22 (m, -CH₂), 114.18 (m, -CF₂), 114.48 (m, -CF₂), 120.17 (td, 3J_C,F= 276, 9.0 Hz, -CF₂), 126.55 (s, Ar-C), 126.75 (s, Ar-C), 128.73(s, Ar-C), 139.97 (s, Ar-C) ppm. ¹⁹F NMR (CDCl₃, 298 K, 565 MHz): δ = -77.71 (m), -78.09 (m), -89.04 (m), -89.41 (s), -89.92 (m) ppm.
³¹P NMR (CDCl₃, 298 K, 243 MHz): δ = -5.30 ppm.
MALDI-TOF MS: for C8H4F12O7P [M-H]^-, calculated: 471.07 Da, found: 471.15 Da. PIPPER-S-Phenylethane was obtained in the same manner as in Example 4, except that 2-phenylethanethiol was used instead of N-(tert-butoxycarbonyl)ethanolamine.
^1H NMR ( _CDCl3 , 298 K, 600 MHz): δ = 1.42 (s, 9H, -Boc), 1.60 (m, 2H, _-CH2 ), 2.96 (m, 2H, _-NHCH2 ), 3.20 (m, 2H, _-NHCH2 ), 4.05 (br, 1H, -NH ₎ _, 4.25 (m, 2H, -OCF2CH2O), 4.44 (m, 2H, -OCF2CH2O) _, _4.76 (br, 1H, -NH) ppm.
^13C NMR ( _CDCl3 , 298 K, 150 MHz): δ = 29.85 (m, _-CH2 ), 40.41 (m, _-CH2 ), 66.22 (m, _-CH2 ), 114.18 (m, _-CF2 ), 114.48 (m, _-CF2 ), 120.17 (td, 3JC _,F = 276, 9.0 Hz, _-CF2 ), 126.55 (s, Ar-C), 126.75 (s, Ar-C), 128.73(s, Ar-C), 139.97 (s, Ar-C) ppm. ^19F NMR ( _CDCl3 , 298 K, 565 MHz): δ = -77.71 (m), -78.09 (m), -89.04 (m), -89.41 (s), -89.92 (m) ppm.
^31P NMR ( _CDCl3 , 298 K, 243 MHz): δ = -5.30 ppm.
MALDI-TOF MS: for C8H4F12O7P [MH] ^- , calculated: 471.07 Da, found: 471.15 Da.

（実施例１０）ＰＩＰＰＥＲ－ＥＧ－ＮＢＤの合成例

(Example 10) Synthesis Example of PIPPER-EG-NBD

実施例４と同様にして合成したＰＩＰＰＥＲ－ＮＨ_２・ＨＣｌ（１２０．０ｍｇ，０．２１８ｍｍｏｌ）及び４－クロロ－７－ニトロベンゾフラザン（4-chloro-7-nitrobenzofurazan；４４．０ｍｇ，０．２２０ｍｍｏｌ）を無水アセトニトリル（５０ｍＬ）に溶解した。この溶液に、１０ｍＬの無水アセトニトリルにトリエチルアミン（ＴＥＡ，２０．０ｍｇ，０．１９８ｍｍｏｌ）を溶解した溶液を、窒素雰囲気下で、滴下することにより加えた。混合液を２５℃で１２時間攪拌し、この反応液を薄層クロマトグラフィー（ＴＬＣ）処理した。溶媒を蒸発させた後、残留物をシリカゲルクロマトグラフィー（φ＝１．５ｃｍ，ｌ＝２０ｃｍ）で精製した。溶離液として、酢酸エチル／ＤＣＭ（１：１）を利用した。これにより、化合物ＰＩＰＰＥＲ－ＥＧ－ＮＢＤを橙色の固体として得た（９６ｍｇ，０．１４４ｍｍｏｌ，収率６５％）。
¹H NMR (CD₃CN, 298 K, 600 MHz): δ = 3.86 (br, 2H, -CH₂), 4.39 (m, 2H, -CH₂), 4.42 (m, 2H, -OCF₂CH₂O), 4.56 (m, 2H, -OCF₂CH₂O), 6.39 (d, ³J = 8.8 Hz, 1H, NBD-CH), 7.46 (br, 1H, -NH), 8.51 (d, ³J = 8.8 Hz, 1H, NBD-CH) ppm.
¹³C NMR (CD₃CN, 298 K, 150 MHz): δ = 44.02 (s, -CH₂), 65.91 (m, -CH₂), 67.26 (s, -CH₂), 100.25 (s, -Ar-C), 114.71 (m, -CF₂), 115.11 (m, -CF₂), 121.83 (td, ³J_C,F = 276, 9.0 Hz, -CF₂), 124.23 (s, Ar-C), 137.71 (s, Ar-C), 145.07 (s, -Ar-C), 145.48 (s), 145.57 (s) ppm.
¹⁹F NMR (CD₃CN, 298 K, 565 MHz): δ = -77.49 (m), -79.02 (m), -87.94 (m), -89.44 (s), -90.89 (m) ppm.
³¹P NMR (CD₃CN, 298 K, 243 MHz): δ = -2.27 ppm.
MALDI-TOF MS: for C₁₆H₁₁F₁₂N₄O₁₀P [M-H⁺], calculated: 676.99 Da, found: 677.66 Da (PIPPER-EG-NBD). PIPPER- _NH2.HCl (120.0 mg, 0.218 mmol) and 4-chloro-7-nitrobenzofurazan (44.0 mg, 0.220 mmol), which were synthesized in the same manner as in Example 4, were dissolved in anhydrous acetonitrile (50 mL). A solution of triethylamine (TEA, 20.0 mg, 0.198 mmol) dissolved in 10 mL of anhydrous acetonitrile was added dropwise to this solution under a nitrogen atmosphere. The mixture was stirred at 25°C for 12 hours, and the reaction solution was subjected to thin layer chromatography (TLC). After evaporating the solvent, the residue was purified by silica gel chromatography (φ=1.5 cm, l=20 cm). As the eluent, ethyl acetate/DCM (1:1) was used. This resulted in the compound PIPPER-EG-NBD being obtained as an orange solid (96 mg, 0.144 mmol, 65% yield).
^1H NMR ( _CD3CN , 298 K, 600 MHz): δ = 3.86 (br, 2H, _-CH2 ), 4.39 (m, 2H, _-CH2 ) _, 4.42 (m, 2H, -OCF2CH2O), 4.56 (m, 2H _, -OCF2CH2O), 6.39 ₍ _d , ³ J = 8.8 Hz, 1H, NBD-CH), 7.46 (br, 1H, -NH), 8.51 (d, ³ J = 8.8 Hz, 1H, NBD-CH) ppm.
^13C NMR ( _CD3CN , 298 K, 150 MHz): δ = 44.02 (s, _-CH2 ), 65.91 (m, _-CH2 ), 67.26 (s, _-CH2 ), 100.25 (s, -Ar-C), 114.71 (m, _-CF2 ), 115.11 (m, _-CF2 ), 121.83 (td, ^3JC _,F = 276, 9.0 Hz, _-CF2 ), 124.23 (s, Ar-C), 137.71 (s, Ar-C), 145.07 (s, -Ar-C), 145.48 (s), 145.57 (s) ppm.
^19F NMR ( _CD3CN , 298 K, 565 MHz): δ = -77.49 (m), -79.02 (m), -87.94 (m), -89.44 (s), -90.89 (m) ppm.
^31P NMR ( _CD3CN , 298K, 243MHz): δ = -2.27ppm.
MALDI-TOF MS: for _{C16H11F12N4O10P} [MH ⁺ _] _, calculated: 676.99 Da _, found: 677.66 Da (PIPPER-EG-NBD ₎ .

（実施例１１）ＰＩＰＰＥＲ－ＤＥＧ－ＮＢＤの合成
ＮＢＤ－ＤＥＧ－ＯＨの合成

(Example 11) Synthesis of PIPPER-DEG-NBD Synthesis of NBD-DEG-OH

４－クロロ－７－ニトロベンゾフラザン（4-chloro-7-nitrobenzofurazan；２．００ｇ，１０．０２ｍｍｏｌ）を窒素雰囲気下で１００ｍＬの乾燥アセトニトリルに溶解し、その溶液を０℃まで冷却した。この溶液に、２０ｍＬの無水アセトニトリルに２－（２－アミノエトキシ）エタノール（2-(2-aminoethoxy)ethanol；２．４０ｇ，２２．８ｍｍｏｌ）を溶解した溶液を、滴下して加え、混合液を室温で１２時間攪拌した。溶媒を乾燥するまで蒸発させ、粗生成物をシリカゲルカラムクロマトグラフィー（φ＝３．０ｃｍ，ｌ＝２０ｃｍ）で精製した。溶離液として、メタノールとジクロロメタンの混合液（ＭｅＯＨ：ＣＨ_２Ｃｌ_２＝１：２０）を用いた。これにより化合物ＮＢＤ－ＤＥＧ－ＯＨ（１．８０ｇ，６．７１ｍｍｏｌ，収率６７％）を得た。
¹H NMR (CD₂Cl₂, 298 K, 600 MHz): δ = 1.97 (s, 1H, -OH), 3.67 (m, 2H, -CH₂), 3.68 (br, 2H, -CH₂), 3.78 (m, 2H, -CH₂), 3.87 (m, 2H, -CH₂), 6.22 (d, ³J = 8.8 Hz, 1H, NBD-CH), 6.95 (br, 1H, -NH), 8.48 (d, ³J = 8.8 Hz, 1H, NBD-CH) ppm.
¹³C NMR (CD₂Cl₂, 298 K, 150 MHz): δ = 43.91, 61.93, 68.37, 72.62, 98.90, 123.97, 136.65, 144.15, 144.24, 144.58.
MALDI-TOF MS: for C₁₀H₁₂N₄O₅[M-H⁺], calculated: 267.07 Da, found: 266.35 Da (NBD-DEG-OH). 4-chloro-7-nitrobenzofurazan (2.00 g, 10.02 mmol) was dissolved in 100 mL of dry acetonitrile under nitrogen atmosphere, and the solution was cooled to 0°C. A solution of 2-(2-aminoethoxy)ethanol (2.40 g, 22.8 mmol) dissolved in 20 mL of anhydrous acetonitrile was added dropwise to the solution, and the mixture was stirred at room temperature for 12 hours. The solvent was evaporated to dryness, and the crude product was purified by silica gel column chromatography (φ=3.0 cm, l=20 cm). A mixture of methanol and dichloromethane (MeOH:CH ₂ Cl ₂ =1:20) was used as the eluent. This gave the compound NBD-DEG-OH (1.80 g, 6.71 mmol, 67% yield).
^1H NMR ( _CD2Cl2 , ₂₉₈ K, 600 MHz): δ = 1.97 (s, 1H, -OH), 3.67 (m, 2H, _-CH2 ), 3.68 (br, 2H, _-CH2 ), 3.78 (m, 2H, _-CH2 ), 3.87 (m, 2H, _-CH2 ), 6.22 (d, ³ J = 8.8 Hz, 1H, NBD-CH), 6.95 (br, 1H, -NH), 8.48 (d, ³ J = 8.8 Hz, 1H, NBD-CH) ppm.
^13C NMR ( _CD2Cl2 , ₂₉₈ K, 150 MHz): δ = 43.91, 61.93, 68.37, 72.62, 98.90, 123.97, 136.65, 144.15, 144.24, 144.58.
MALDI-TOF MS: for _C10H12N4O5 [MH ⁺ _] _, calculated: 267.07 Da _, found: 266.35 Da (NBD-DEG-OH).

ＰＩＰＰＥＲ－ＤＥＧ－ＮＢＤの合成

Synthesis of PIPPER-DEG-NBD

実施例１と同様にして合成したＰＩＰＰＥＲ－Ｃｌの粗生成物を２００ｍＬの無水トルエンに溶解した。その後、１．０ｍＬのピリジンと前記で合成したＮＢＤ－ＤＥＧ－ＯＨ（１．５０ｇ，５．５９ｍｍｏｌ）を窒素雰囲気下で加えた。この混合物を８０℃で１時間加熱した後、反応物を薄層クロマトグラフィーＴＬＣ処理した。溶媒を蒸発させた後、残留物をジクロロメタン（ＤＣＭ；１００ｍＬ）で抽出し、Milli-Q^TM 水（１５０ｍＬ）で洗浄した。有機層を無水硫酸ナトリウムで乾燥し、溶媒を除去した。粗生成物をシリカゲルクロマトグラフィー（φ＝３．５ｃｍ，ｌ＝２０ｃｍ）で精製した（Ｒｆ＝０．２０，ＳｉＯ_２）。溶離液としてメタノールとジクロロメタンの混合液（ＭｅＯＨ：ＤＣＭ＝１：２０）を使用した。これにより化合物ＰＩＰＰＥＲ－ＤＥＧ－ＮＢＤを黄色の固体として得た（１．５５ｇ，２．１５ｍｍｏｌ，収率３８％）。
¹H NMR (CD₂Cl₂, 298 K, 600 MHz): δ = 3.73 (br, 2H, -CH₂), 3.80 (m, 2H, -CH₂), 3.87 (m, 2H, -CH₂), 4.32 (dd, ³J = 8.4, 5.4 Hz, 2H, ,-CH₂), 4.36 (m, 2H, -OCF₂CH₂O), 4.55 (m, 2H, -OCF₂CH₂O), 6.52 (d, ³J = 8.8 Hz, 1H, NBD-CH), 7.37 (t, ³J = 5.4 Hz, 1H, -NH), 8.48 (d, ³J = 8.8 Hz, 1H, NBD-CH) ppm.
¹³C NMR (CD₂Cl₂, 298 K, 150 MHz): δ = 44.30 (s, -CH₂), 65.97 (d, ³J_C,F = 4.5 Hz, -CH₂), 68.57 (s, -CH₂), 68.60 (s, -CH₂), 70.12 (m, -CH₂), 99.49 (s, -Ar-C), 114.49 (m, -CF₂), 114.81 (m, -CF₂), 121.15 (td, ³J_C,F = 276, 9.0 Hz, -CF₂), 124.18 (s, Ar-C), 136.84 (s, Ar-C), 144.60 (s, -Ar-C), 144.87 (s, 2C) ppm.
¹⁹F NMR (CD₂Cl₂, 298 K, 565 MHz): δ = -78.09 (m), -79.07 (m), -88.72 (m), -89.73 (s), -90.67 (m) ppm.
³¹P NMR (CD₂Cl₂, 298 K, 243 MHz): δ = -2.56 ppm.
MALDI-TOF MS: for C₁₈H₁₅F₁₂N₄O₁₁P [M+H⁺], calculated: 723.30 Da, found: 723.89 Da (PIPPER-DEG-NBD). The crude product of PIPPER-Cl, synthesized in the same manner as in Example 1, was dissolved in 200 mL of anhydrous toluene. Then, 1.0 mL of pyridine and the NBD-DEG-OH (1.50 g, 5.59 mmol) synthesized above were added under a nitrogen atmosphere. The mixture was heated at 80° C. for 1 h, and the reaction was then subjected to thin layer chromatography (TLC). After evaporation of the solvent, the residue was extracted with dichloromethane (DCM; 100 mL) and washed with Milli-Q ^TM water (150 mL). The organic layer was dried over anhydrous sodium sulfate and the solvent was removed. The crude product was purified by silica gel chromatography (φ=3.5 cm, l=20 cm) (Rf=0.20, SiO ₂ ). A mixture of methanol and dichloromethane (MeOH:DCM=1:20) was used as the eluent. This resulted in the compound PIPPER-DEG-NBD as a yellow solid (1.55 g, 2.15 mmol, 38% yield).
^1H NMR ( _CD2Cl2 , ₂₉₈ K, 600 MHz): δ = 3.73 (br, 2H, _-CH2 ), 3.80 (m, 2H, _-CH2 ), 3.87 (m, 2H, _-CH2 ), 4.32 (dd, ³ J = 8.4, 5.4 Hz, 2H, , _-CH2 ) _, 4.36 (m, 2H, -OCF2CH2O), _4.55 (m, 2H, -OCF2CH2O), 6.52 (d, ³ J = 8.8 Hz, 1H, NBD-CH), 7.37 (t, ³ J = 5.4 _Hz ^{, 1H, -NH), 8.48 (d, 3} _J = 8.8 Hz, 1H, NBD-CH) ppm.
^13C NMR ( _CD2Cl2 , ₂₉₈ K, 150 MHz): δ = 44.30 (s, _-CH2 ), 65.97 (d, ^3JC _,F = 4.5 Hz, _-CH2 ), 68.57 (s, -CH2) _, 68.60 (s, _-CH2 ), 70.12 (m, _-CH2 ), 99.49 (s, -Ar-C), 114.49 (m, _-CF2 ), 114.81 (m, _-CF2 ), 121.15 (td, ^3JC _,F = 276, 9.0 Hz, _-CF2 ), 124.18 (s, Ar-C), 136.84 (s, Ar-C), 144.60 (s, -Ar-C), 144.87 (s, 2C) ppm.
^19F NMR ( _CD2Cl2 , ₂₉₈ K, 565 MHz): δ = -78.09 (m), -79.07 (m), -88.72 (m), -89.73 (s), -90.67 (m) ppm.
^31P NMR ( _CD2Cl2 , _298K , 243MHz): δ = -2.56 ppm.
MALDI-TOF MS: for _{C18H15F12N4O11P} [ _M +H ⁺ ] _, calculated: 723.30 Da, _found : 723.89 Da (PIPPER-DEG-NBD ₎ .

（実施例１２）ＰＩＰＰＥＲ－ＴＥＧ－ＮＢＤの合成
ＮＢＤ－ＴＥＧ－ＯＨの合成

(Example 12) Synthesis of PIPPER-TEG-NBD Synthesis of NBD-TEG-OH

４－クロロ－７－ニトロベンゾフラザン（4-chloro-7-nitrobenzofurazan；１．２０ｇ，６．０１ｍｍｏｌ）を窒素雰囲気下で、１００ｍＬの乾燥アセトニトリルに溶解し、この溶液を０℃まで冷却した。この反応溶液に、３０ｍＬの無水アセトニトリルに２－［２－（２－アミノエトキシ）エトキシ］エタノール（１．００ｇ，６．７０ｍｍｏｌ）を溶解した溶液を滴下により加え、混合物を室温で１２時間攪拌した。溶媒を蒸発させて乾燥し、得られた粗生成物をシリカゲルカラムクロマトグラフィー（φ＝３．０ｃｍ，ｌ＝２０ｃｍ）で精製した。溶離液としてメタノールとジクロロメタンの混合液（ＭｅＯＨ：ＣＨ_２Ｃｌ_２＝１：２０）を用いた。これにより目的の化合物ＮＢＤ－ＴＥＧ－ＯＨ（１．６５ｇ，５．２９ｍｍｏｌ，収率７９％）を得た。 4-chloro-7-nitrobenzofurazan (1.20 g, 6.01 mmol) was dissolved in 100 mL of dry acetonitrile under a nitrogen atmosphere, and the solution was cooled to 0°C. A solution of 2-[2-(2-aminoethoxy)ethoxy]ethanol (1.00 g, 6.70 mmol) dissolved in 30 mL of anhydrous acetonitrile was added dropwise to the reaction solution, and the mixture was stirred at room temperature for 12 hours. The solvent was evaporated to dryness, and the obtained crude product was purified by silica gel column chromatography (φ=3.0 cm, l=20 cm). A mixture of methanol and dichloromethane (MeOH:CH ₂ Cl ₂ =1:20) was used as the eluent. As a result, the target compound NBD-TEG-OH (1.65 g, 5.29 mmol, 79% yield) was obtained.

ＰＩＰＰＥＲ－ＴＥＧ－ＮＢＤの合成

Synthesis of PIPPER-TEG-NBD

実施例１１で用いたＮＢＤ－ＤＥＧ－ＯＨを前記で合成したＮＢＤ－ＴＥＧ－ＯＨに置き換えた以外は、同様にして、褐色の液状のＰＩＰＰＥＲ－ＴＥＧ－ＮＢＤを得た（１．２７ｇ，１．６６ｍｍｏｌ，収率３２％）。
¹H NMR (CD₂Cl₂, 298 K, 600 MHz): δ = 3.69 (m, 6H, -CH₂), 3.73 (m, 2H, -CH₂), 3.84 (m, 2H, -CH₂), 4.30 (m, 2H, -CH₂), 4.38(m, 2H, -OCF₂CH₂O), 4.56 (m, 2H, -OCF₂CH₂O), 6.22 (d, ³J = 8.8 Hz, 1H, NBD-CH), 7.20 (br, 1H, -NH), 8.47 (d, ³J = 8.8 Hz, 1H, NBD-CH) ppm.
¹³C NMR (CD₂Cl₂, 298 K, 150 MHz): δ = 44.17 (s, -CH₂), 65.79 (m, -CH₂), 68.58 (s, -CH₂), 69.02 (m, -CH₂), 70.19 (m, -CH₂), 70.92 (m, -2C), 99.31 (s, -Ar-C), 114.52 (m, -CF₂), 114.82 (m, -CF₂), 121.28 (td, ³J_C,F = 276, 9.0 Hz, -CF₂), 124.04 (s, Ar-C), 137.00 (s, Ar-C), 144.54 (s, -Ar-C), 144.75 (s, -Ar-C) 144.91 (s, -Ar-C)ppm.
¹⁹F NMR (CD₂Cl₂, 298 K, 565 MHz): δ = -78.20 (m), -79.02 (m), -88.90 (m), -89.76 (s), -90.56 (m) ppm.
³¹P NMR (CD₂Cl₂, 298 K, 243 MHz): δ = -2.78 ppm.
MALDI-TOF MS: for C₂₀H₁₈F₁₂N₄O₁₂P [M-H⁺], calculated: 765.05 Da, found: 764.97 Da (PIPPER-TEG-NBD). A brown liquid PIPPER-TEG-NBD was obtained in the same manner as in Example 11 except that the NBD-DEG-OH used in Example 11 was replaced with the NBD-TEG-OH synthesized above (1.27 g, 1.66 mmol, yield 32%).
^1H NMR ( _CD2Cl2 , ₂₉₈ K, 600 MHz): δ = 3.69 (m, 6H, _-CH2 ), 3.73 (m, 2H, _-CH2 ), 3.84 (m, 2H, _-CH2 ) _, 4.30 (m, 2H, _-CH2 ) _, 4.38(m, 2H, -OCF2CH2O) _, 4.56 (m, 2H, -OCF2CH2O), 6.22 (d, ³ J = 8.8 Hz, 1H, NBD-CH), _7.20 (br, 1H, -NH), 8.47 (d, ³ J = 8.8 Hz, 1H, NBD-CH) ppm.
^13C NMR ( _CD2Cl2 , ₂₉₈ K, 150 MHz): δ = 44.17 (s, _-CH2 ), 65.79 (m, _-CH2 ), 68.58 (s, _-CH2 ), 69.02 (m, -CH2), 70.19 (m, _-CH2 ), 70.92 (m, _-2C ), 99.31 (s, -Ar-C), 114.52 (m, _-CF2 ), 114.82 (m, _-CF2 ), 121.28 (td, ^3JC _,F = 276, 9.0 Hz, _-CF2 ), 124.04 (s, Ar-C), 137.00 (s, Ar-C), 144.54 (s, -Ar-C), 144.75 (s, -Ar-C) 144.91 (s, -Ar-C)ppm.
^19F NMR ( _CD2Cl2 , ₂₉₈ K, 565 MHz): δ = -78.20 (m), -79.02 (m), -88.90 (m), -89.76 (s), -90.56 (m) ppm.
^31P NMR ( _CD2Cl2 , _298K , 243MHz): δ = -2.78 ppm.
MALDI-TOF MS: for _{C20H18F12N4O12P} [MH ₊ _] _, ^calculated : 765.05 Da, found: 764.97 Da (PIPPER-TEG-NBD ₎ .

（実施例１３）PIPPER-Fmocの合成

(Example 13) Synthesis of PIPPER-Fmoc

実施例１と同様にして合成したＰＩＰＰＥＲ－Ｃｌの粗生成物を１００ｍＬの無水トルエンに溶解した。その後、この溶液に、１．０ｍＬのピリジンと２－（Ｆｍｏｃ－アミノ）エタノール（2-(Fmoc-amino)ethanol；２．００ｇ，７．０６ｍｍｏｌ）を窒素雰囲気下で加えた。混合物を１時間還流した後、反応物を薄層クロマトグラフィーＴＬＣ処理した。溶媒を蒸発させた後、残留物をジクロロメタン（ＤＣＭ；１００ｍＬ）で抽出し、Milli-Q^TM 水（１５０ｍＬ）で洗浄した．有機層を無水硫酸ナトリウムで乾燥し、溶媒を除去した。粗生成物をシリカゲルクロマトグラフィー（φ＝３．５ｃｍ，ｌ＝２０ｃｍ）で精製した。溶離液として、酢酸エチルとジクロロメタンとの混合液（酢酸エチル：ＤＣＭ＝２：８）を用いた。これにより、無色の液体として化合物ＰＩＰＰＥＲ－Ｆｍｏｃ（２．５０ｇ，３．３９ｍｍｏｌ，収率４８％）を得た。
¹H NMR (CDCl₃, 298 K, 600 MHz): δ = 3.49 (dd, ³J = 10.8, 5.4 Hz, 2H, CH₂), 4.18 (dd, ³J = 10.8, 5.4 Hz, 2H, CH₂), 4.20 (t, ³J = 7.2 Hz, 1H), 4.28 (m, 2H, -OCF₂CH₂O), 4.43 (d, ³J = 7.2 Hz, 2H), 4.48 (m, 2H, -OCF₂CH₂O), 5.33 (t, ³J = 6.0 Hz, 1H, N-H), 7.31 (td, ³J = 7.8 Hz, ⁵J = 0.8 Hz, 2H, Ar-CH), 7.40 (t, ³J = 7.8 Hz, 2H, Ar-CH), 7.58 (d, ³J = 7.8 Hz, 2H, Ar-CH), 7.77 (d, ³J = 7.8 Hz, 2H, Ar-CH) ppm.
¹³C NMR (CDCl₃, 298 K, 150 MHz): δ = 41.28 (s, -CH₂), 47.34 (d, ³J_C,F = 4.5 Hz, -CH₂), 65.68 (m, -CH₂), 67.11 (s, -CH₂), 68.37 (m, -CH₂), 114.29 (m, -CF₂), 114.61 (m, -CF₂), 120.26 (d, ³J_C,F = 50.16 Hz, Ar), 120.72 (td, ³J_C,F = 276, 9.0 Hz, -CF₂), 125.12 (d, ³J_C,F = 69.66 Hz, Ar), 127.27 (d, ³J_C,F = 98.4 Hz, Ar), 127.98 (d, ³J_C,F = 97.8 Hz, Ar), 141.55 (s, Ar), 138.25 (s, Ar-C), 143.97 (s, -Ar-C), 156.6 (s, C=O) ppm.
¹⁹F NMR (CDCl₃, 298 K, 565 MHz): δ = -77.50 (m), -78.85 (m), -88.01 (m), -89.39 (s), -90.69 (m) ppm.
³¹P NMR (CDCl₃, 298 K, 243 MHz): δ = -2.42 ppm.
MALDI-TOF MS: for C₂₅H₂₀F₁₂NO₉P.Na [M+Na]⁺, calculated: 760.38 Da, found: 760.148 Da (PIPPER-Fmoc+Na⁺); C₂₅H₂₀F₁₂NO₉P.K [M+K]⁺, calculated: 776.49 Da, found: 776.165 Da (PIPPER-Fmoc+K⁺).
The crude product of PIPPER-Cl synthesized in the same manner as in Example 1 was dissolved in 100 mL of anhydrous toluene. Then, 1.0 mL of pyridine and 2-(Fmoc-amino)ethanol (2.00 g, 7.06 mmol) were added to this solution under nitrogen atmosphere. The mixture was refluxed for 1 hour, and the reaction product was treated by thin layer chromatography (TLC). After evaporating the solvent, the residue was extracted with dichloromethane (DCM; 100 mL) and washed with Milli-Q ^TM water (150 mL). The organic layer was dried over anhydrous sodium sulfate, and the solvent was removed. The crude product was purified by silica gel chromatography (φ=3.5 cm, l=20 cm). A mixture of ethyl acetate and dichloromethane (ethyl acetate:DCM=2:8) was used as the eluent. This gave the compound PIPPER-Fmoc (2.50 g, 3.39 mmol, 48% yield) as a colorless liquid.
^1H NMR ( _CDCl3 , 298 K, 600 MHz): δ = 3.49 (dd, ³ J = 10.8, 5.4 Hz, 2H, _CH2 ), 4.18 (dd, ³ J = 10.8, 5.4 Hz, 2H, _CH2 ), 4.20 (t, ³ J = 7.2 Hz, 1H), 4.28 (m, 2H, _-OCF2CH2O ), 4.43 ₍ d, ³ J = 7.2 Hz, 2H), 4.48 (m, 2H _, _-OCF2CH2O ), 5.33 (t, ³ J = 6.0 Hz, 1H, NH), 7.31 (td, ³ J = 7.8 Hz, ⁵ J = 0.8 Hz, 2H, Ar-CH), 7.40 (t, ³ J = 7.8 Hz, 2H, Ar-CH), 7.58 (d, ³ J = 7.8 Hz, 2H, Ar-CH), 7.77 (d, ³ J = 7.8 Hz, 2H, Ar-CH) ppm.
^13C NMR ( _CDCl3 , 298 K, 150 MHz): δ = 41.28 (s, _-CH2 ), 47.34 (d, ^3JC _,F = 4.5 Hz, _-CH2 ), 65.68 (m, _-CH2 ), 67.11 (s, _-CH2 ), 68.37 (m, _-CH2 ), 114.29 (m, -CF2), 114.61 (m, _-CF2 ), 120.26 (d, _3JC _,F = 50.16 Hz, Ar), 120.72 (td, ^3JC _,F = 276 ^, 9.0 Hz, _-CF2 ), 125.12 (d, ^3JC _,F = 69.66 Hz, Ar), 127.27 (d, ³ J _C,F = 98.4 Hz, Ar), 127.98 (d, ³ J _C,F = 97.8 Hz, Ar), 141.55 (s, Ar), 138.25 (s, Ar-C), 143.97 (s, -Ar-C), 156.6 (s, C=O) ppm.
^19F NMR ( _CDCl3 , 298 K, 565 MHz): δ = -77.50 (m), -78.85 (m), -88.01 (m), -89.39 (s), -90.69 (m) ppm.
^31P NMR ( _CDCl3 , 298 K, 243 MHz): δ = -2.42 ppm.
MALDI-TOF MS: for C _{25 H 20 F 12 NO 9 P.Na [M+Na] + , calculated: 760.38 Da, found: 760.148 Da (PIPPER-Fmoc+Na + ); C 25} _{H 20} _F ₁₂ ^NO ⁹ _PK _[ _M ₊ K] ⁺ , calculated: 776.49 Da, found: 776.165 Da (PIPPER-Fmoc+K ⁺ ).

Claims

The following formula (I):

(In the formula, R represents a hydroxyl group, a chlorine atom, or a group represented by the formula: -O- (organic group), -NH- (organic group), or -S- (organic group), and n represents an integer of 1 to 7.)
A compound represented by the formula:

The compound according to claim 1, wherein in formula (I), R is a chlorine atom.

The compound according to claim 1, wherein in formula (I), R is a hydroxyl group.

The compound according to claim 1, wherein in formula (I), R is a group represented by the formula: -O- (organic group), -NH- (organic group), or -S- (organic group), and the organic group contains at least one alkyne group or maleimide group.

The compound according to claim 1, wherein in formula (I), R is a group represented by the formula: -O- (organic group), -NH- (organic group), or -S- (organic group), and the organic group includes a group derived from an active ingredient of a pharmaceutical or diagnostic agent.

The compound according to claim 1, wherein in formula (I), R is a group represented by the formula: -O- (organic group), -NH- (organic group), or -S- (organic group), and the organic group contains a light-emitting and/or light-absorbing functional group.

The following formula (II):
_HOCH2CF2O- ₍ _CF2CF2O ) _n- _CF2CH2OH ₍ _II )
(In the formula, n has the same meaning as in formula (I) described in claim 1.)
3. A process for preparing the compound of claim 2, comprising reacting a fluorinated polyethylene glycol of the formula: with phosphoryl chloride.

A method for producing the compound of claim 3, comprising reacting the compound of claim 2 with water.

A method for producing a compound represented by formula (I) in which R is a group represented by the formula: -O- (organic group), _-NH- (organic group) or -S- (organic group), comprising reacting the compound according to claim 2 with a compound represented by the formula: HO- (organic group), H 2 N- (organic group) or HS- (organic group).

A method for producing the compound according to claim 5 or 6, comprising subjecting the compound according to claim 4 to a click reaction with a reaction reagent.

A composition comprising the compound according to any one of claims 1 to 6 and an aqueous solvent.

The composition according to claim 11, further comprising an inorganic ion.

An inorganic ion inclusion agent comprising the compound according to any one of claims 1 to 6.