JP2006257069A

JP2006257069A - Methylene substitution product and alkenyl phosphate substitution product of fluorescence-labeled sphingosine 1-phosphate

Info

Publication number: JP2006257069A
Application number: JP2005281750A
Authority: JP
Inventors: Shigeo Katsumura; 成雄勝村; Hiroko Hasegawa; 浩子長谷川; Tetsuya Yamamoto; 哲也山本
Original assignee: KEMIKUREA KK; Kwansei Gakuin Educational Foundation; Chemicrea Inc
Current assignee: KEMIKUREA KK; Kwansei Gakuin Educational Foundation; Chemicrea Inc
Priority date: 2005-02-17
Filing date: 2005-09-28
Publication date: 2006-09-28

Abstract

<P>PROBLEM TO BE SOLVED: To provide a sphingosine analog having a fluorescence-labeled site, useful for elucidating a location or the metabolic mechanism of sphingosine in cells and scarcely hydrolyzing a phosphoric ester moiety. <P>SOLUTION: A fluorescence-labeled sphingosine 1-phosphate methylene substitution product is represented by general formula (I) (wherein, R<SP>1</SP>and R<SP>2</SP>are each a hydrogen atom or a lower alkyl group; Q<SP>1</SP>and Q<SP>2</SP>are each a hydrogen atom or a protecting group of amino group; Q<SP>3</SP>is a hydrogen atom or a protecting group of hydroxy group; X is a 1-15C alkylene group; Z is a fluorescence-labeling group; and Q<SP>1</SP>and Q<SP>2</SP>, Q<SP>2</SP>and Q<SP>3</SP>or Q<SP>1</SP>and Q<SP>3</SP>together each may form a ring) or an acceptable salt thereof. <P>COPYRIGHT: (C)2006,JPO&NCIPI

Description

本発明はスフィンゴ脂質の細胞内での挙動を解明するために有用な、蛍光標識基を導入したスフィンゴシン類縁体に関するものである。 The present invention relates to a sphingosine analog having a fluorescent labeling group useful for elucidating the intracellular behavior of sphingolipids.

スフィンゴミエリンの代謝物であるスフィンゴ脂質は、細胞死、細胞増殖、細胞分化、プロテインキナーゼＣ阻害など様々な生体内情報伝達に関与していることから大きな注目を集めており、スフィンゴ脂質が関係する生体現象の解明が多くの研究者により積極的に行われている。 Sphingolipid, which is a metabolite of sphingomyelin, has attracted a great deal of attention because it is involved in various in vivo signal transduction such as cell death, cell proliferation, cell differentiation, and protein kinase C inhibition. Biological phenomena have been elucidated actively by many researchers.

そのスフィンゴ脂質の１つであるスフィンゴシンは主に酵母細胞に存在するが、これは哺乳動物細胞においてシグナル分子として知られているスフィンゴシンに対応しており、多くの研究が行われている。しかしながら、スフィンゴシンの細胞内での存在場所や発現に至る経路、作用機序などの情報は充分に得られていない。 One of the sphingolipids, sphingosine, is mainly present in yeast cells. This corresponds to sphingosine known as a signal molecule in mammalian cells, and many studies have been conducted. However, information on the location of sphingosine in the cell, the route leading to its expression, the mechanism of action, etc. has not been sufficiently obtained.

現在、この代謝機構を解明するものとして、Ｃ６−ＮＢＤフィトスフィンゴシンやＣ１２−ＮＢＤフィトスフィンゴシンという化合物が市販されているが、これらは副鎖であるアシル基に蛍光標識基ユニットがあり、この部分は加水分解酵素により切断されるため、フィトスフィンゴシンが代謝されたフィトスフィンゴシン−１−リン酸の細胞内の局在場所や代謝物の挙動を解明するためには有用ではない。 At present, as a mechanism for elucidating this metabolic mechanism, compounds such as C6-NBD phytosphingosine and C12-NBD phytosphingosine are commercially available, and these have a fluorescent labeling group unit in the acyl group as a sub-chain. Since it is cleaved by the hydrolase, it is not useful for elucidating the intracellular location of phytosphingosine-1-phosphate in which phytosphingosine is metabolized and the behavior of the metabolite.

このような問題を解決するために代謝をうけない主鎖に蛍光標識基を導入した化合物も開発されている。例えば、特許文献１には、スフィンゴシン類縁体のアルキル基末端に蛍光標識基を導入した化合物が記載されている。また、特許文献２には、フィトスフィンゴシン類縁体のアルキル基末端に蛍光標識基を導入した化合物が記載されている。
特開２００３−２６１７９４号公報特開２００４−２６９３８１号公報 In order to solve such a problem, a compound in which a fluorescent labeling group is introduced into a main chain that is not metabolized has been developed. For example, Patent Document 1 describes a compound in which a fluorescent labeling group is introduced at the end of an alkyl group of a sphingosine analog. Patent Document 2 describes a compound in which a fluorescent labeling group is introduced at the end of an alkyl group of a phytosphingosine analog.
JP 2003-261794 A JP 2004-269281 A

しかしながら、特許文献１および２に記載されている化合物は細胞内でリン酸エステル部分が加水分解されるため、受容体との複合体の挙動観測が困難であるという問題を有している。 However, the compounds described in Patent Documents 1 and 2 have a problem that it is difficult to observe the behavior of the complex with the receptor because the phosphate ester moiety is hydrolyzed in the cell.

本発明は上記事情に鑑みなされたものであり、スフィンゴシンの細胞内での存在場所や代謝機構を解明するために有用な、リン酸エステル部分が加水分解されにくい蛍光標識基部位を有するスフィンゴシン類縁体を提供することを目的とするものである。 The present invention has been made in view of the above circumstances, and is useful for elucidating the location and metabolic mechanism of sphingosine in cells, and a sphingosine analog having a fluorescent labeling group site where the phosphate ester moiety is difficult to hydrolyze Is intended to provide.

本発明の蛍光標識化スフィンゴシン１−リン酸メチレン置換体は、下記一般式（Ｉ）

The fluorescently labeled sphingosine 1-methylene phosphate-substituted product of the present invention has the following general formula (I)

（式中、Ｒ^１およびＲ^２は水素原子または低級アルキル基であり、Ｑ^１およびＱ^２は水素原子またはアミノ基の保護基であり、Ｑ^３は水素原子または水酸基の保護基であり、Ｘは炭素数１〜１５のアルキレン基であり、Ｚは蛍光標識基である。Ｑ^１とＱ^２、Ｑ²とＱ^３あるいはＱ^１とＱ^３はそれぞれ一体となって環を形成していてもよい。）で表されることを特徴とするものである。 Wherein R ¹ and R ² are a hydrogen atom or a lower alkyl group, Q ¹ and Q ² are a hydrogen atom or an amino group protecting group, Q ³ is a hydrogen atom or a hydroxyl protecting group, Is an alkylene group having 1 to 15 carbon atoms and Z is a fluorescent labeling group, Q ¹ and Q ² , Q ² and Q ^3, or Q ¹ and Q ³ may be combined to form a ring. It is characterized by being expressed by

上記一般式（Ｉ）で表される化合物は、塩基であるアミン部位と酸であるリン酸部位を分子内に含んでいるため、溶液中のｐＨや単離する時のｐＨによって、アミノ酸類のように分子内塩や溶液中の酸・塩基との分子間塩を形成することがあるが、本発明の蛍光標識化スフィンゴシン１−リン酸メチレン置換体はこのような許容される塩も包むものである。これらの許容される分子間塩としては、塩酸塩、硝酸塩、硫酸塩、リン酸塩、酢酸塩、トリフルオロ酢酸塩などの酸性塩、ナトリウム塩、リチウム塩、カリウム塩、カルシウム塩、マグネシウム塩、バリウム塩などの塩基性塩が挙げられる。 Since the compound represented by the above general formula (I) contains an amine site as a base and a phosphate site as an acid in the molecule, depending on the pH in the solution and the pH at the time of isolation, In this way, an intramolecular salt or an intermolecular salt with an acid / base in a solution may be formed, but the fluorescently labeled sphingosine 1-phosphate methylene phosphate of the present invention also encompasses such an acceptable salt. . These acceptable intermolecular salts include hydrochlorides, nitrates, sulfates, phosphates, acetates, trifluoroacetates and other acidic salts, sodium salts, lithium salts, potassium salts, calcium salts, magnesium salts, Examples include basic salts such as barium salts.

本発明の蛍光標識化スフィンゴシン１−リン酸メチレン置換体は、蛍光標識基を代謝を受けないスフィンゴシンの主鎖に導入しているため、フィトスフィンゴシン−１−リン酸の細胞内の局在場所や代謝物の挙動を解明するのに有用である。また、本発明の蛍光標識化スフィンゴシン１−リン酸メチレン置換体はリン酸エステル部分のリンと主鎖の炭素の結合が強固であって、細胞内でリン酸エステル部分が加水分解されにくいため、フィトスフィンゴシン−１−リン酸の受容体との複合体の挙動を解明するのにも有用である。 Since the fluorescently labeled sphingosine 1-phosphate methylene substitute of the present invention introduces a fluorescent labeling group into the main chain of sphingosine that does not undergo metabolism, the location of phytosphingosine-1-phosphate in the cell It is useful for elucidating the behavior of metabolites. In addition, since the fluorescently labeled sphingosine 1-phosphate methylene phosphate of the present invention has a strong bond between phosphorus in the phosphate ester moiety and carbon in the main chain, and the phosphate ester moiety is not easily hydrolyzed in the cell, It is also useful for elucidating the behavior of the complex with the receptor for phytosphingosine-1-phosphate.

（式中、Ｒ^１およびＲ^２は水素原子または低級アルキル基であり、Ｑ^１およびＱ^２は水素原子またはアミノ基の保護基であり、Ｑ^３は水素原子または水酸基の保護基であり、Ｘは炭素数１〜１５のアルキレン基であり、Ｚは蛍光標識基である。Ｑ^１とＱ^２、Ｑ²とＱ^３あるいはＱ^１とＱ^３はそれぞれ一体となって環を形成していてもよい。）で表されることを特徴とする。 Wherein R ¹ and R ² are a hydrogen atom or a lower alkyl group, Q ¹ and Q ² are a hydrogen atom or an amino group protecting group, Q ³ is a hydrogen atom or a hydroxyl protecting group, Is an alkylene group having 1 to 15 carbon atoms and Z is a fluorescent labeling group, Q ¹ and Q ² , Q ² and Q ^3, or Q ¹ and Q ³ may be combined to form a ring. It is characterized by being expressed by

本発明における低級アルキル基とは、枝分かれを有していてもよい炭素数１から８のアルキル基であり、メチル基、エチル基、プロピル基、イソプロピル基、ブチル基、ｓｅｃ−ブチル基、ｔ−ブチル基、ペンチル基等を例示することができる。 The lower alkyl group in the present invention is an alkyl group having 1 to 8 carbon atoms which may have a branch, and includes a methyl group, an ethyl group, a propyl group, an isopropyl group, a butyl group, a sec-butyl group, t- A butyl group, a pentyl group, etc. can be illustrated.

本発明のアミノ基の保護基としては、公知のアミノ基の保護基がいずれも使用でき、たとえばメトキシカルボニル基、ｔ−ブトキシキシカルボニル基、ベンジルオキシカルボニル基、アセチル基、トリフルオロアセチル基、ベンジル基、p-メトキシベンジル基、p-メトキシフェニル基等を挙げることができる。 As the amino-protecting group of the present invention, any known amino-protecting group can be used, for example, methoxycarbonyl group, t-butoxyoxycarbonyl group, benzyloxycarbonyl group, acetyl group, trifluoroacetyl group, benzyl group. Group, p-methoxybenzyl group, p-methoxyphenyl group and the like.

本発明の水酸基の保護基としては、公知の水酸基の保護基がいずれも使用でき、たとえば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、メトキシメチル基、テトラヒドロピラニル基、テトラヒドロフラニル基、ｐ−メトキシベンジル基、
等を例示することができる。 As the hydroxyl-protecting group of the present invention, any known hydroxyl-protecting group can be used. For example, trimethylsilyl group, triethylsilyl group, t-butyldimethylsilyl group, methoxymethyl group, tetrahydropyranyl group, tetrahydrofuranyl group, p-methoxybenzyl group,
Etc. can be illustrated.

Ｑ^１とＱ^２、Ｑ²とＱ^３あるいはＱ^１とＱ^３はそれぞれ一体となって環を形成していてもよく、Ｑ^１とＱ^２が一体となって環を形成している保護基の例としては、ジフェニルマレイミド環、フタルイミド環、ジチアコハク酸イミド環、テトラメチルジシラザシクロペンタン環、テトラメチルジシライソインドリン環等を例示することができる。 Q ¹ and Q ² , Q ² and Q ³ or Q ¹ and Q ³ may be combined to form a ring, and Q ¹ and Q ² are combined to form a ring. Examples of these include diphenylmaleimide ring, phthalimide ring, dithiasuccinimide ring, tetramethyldisilazacyclopentane ring, tetramethyldisilaisoindoline ring, and the like.

Ｑ²とＱ^３、あるいはＱ^１とＱ^３が一体となって環を形成している保護基の例としては、オキサゾリジンノン環、ジメチルオキサゾリジン環等を例示することができる。 Examples of the protecting group in which Q ² and Q ³ or Q ¹ and Q ³ form a ring together include an oxazolidinenon ring and a dimethyloxazolidine ring.

本発明の蛍光標識基とは、アミノ基と結合することが可能な蛍光標識ユニットであれば何ら限定されることはなく、たとえば、７−ニトロベンゾ−２−オキサ−１，３−ジアゾール基、７−ニトロベンゾ［１，２，５］オキサジアゾール基、アントラセニル基、ダンシル基等を例示することができる。 The fluorescent labeling group of the present invention is not particularly limited as long as it is a fluorescent labeling unit capable of binding to an amino group. For example, a 7-nitrobenzo-2-oxa-1,3-diazole group, 7 -Nitrobenzo [1,2,5] oxadiazole group, anthracenyl group, dansyl group and the like can be exemplified.

本発明の一般式（Ｉ）で表される化合物は、公知の化合物からたとえば下記に示すメタセシス反応を利用し、水酸基やアミノ基を保護・脱保護することにより容易に製造することができる。

The compound represented by the general formula (I) of the present invention can be easily produced from a known compound by protecting / deprotecting a hydroxyl group or an amino group using, for example, the following metathesis reaction.

メタセシス反応には、既存の各種メタセシス触媒が好適に用いられるが、中でもルテニウムカルベン錯体が、反応効率の点で好ましい。 Various existing metathesis catalysts are suitably used for the metathesis reaction, and among them, a ruthenium carbene complex is preferable in terms of reaction efficiency.

メタセシス反応は反応に関与しない溶媒中で行うことが好ましく、ベンゼン、トルエン、キシレン、ヘキサン、シクロヘキサン等の炭化水素系溶媒、テトラヒドロフラン、ジメトキシエタン、ジオキサン等のエーテル系溶媒、ジクロロメタン、クロロホルム、ジクロロエタン等のハロゲン化溶媒等を例示することができる。 The metathesis reaction is preferably carried out in a solvent that does not participate in the reaction, hydrocarbon solvents such as benzene, toluene, xylene, hexane, cyclohexane, ether solvents such as tetrahydrofuran, dimethoxyethane, dioxane, dichloromethane, chloroform, dichloroethane, etc. A halogenated solvent etc. can be illustrated.

メタセシス反応の反応温度は、0℃〜150℃の温度範囲から適宜選択することができるが、反応速度ならびに経済的観点から室温（20℃）〜80℃の範囲が好ましい。
以下、本発明を実施例および参考例によりさらに詳しく説明する。 The reaction temperature of the metathesis reaction can be appropriately selected from a temperature range of 0 ° C. to 150 ° C., but the range of room temperature (20 ° C.) to 80 ° C. is preferable from the viewpoint of reaction rate and economics.
Hereinafter, the present invention will be described in more detail with reference to Examples and Reference Examples.

（参考例１）

(Reference Example 1)

11-ヒドロキシ-1-ウンデセン(10.0 ml, 49.9 mmol)のTHF(100 ml)溶液に、0 ℃でトリエチルアミン(17.4 ml, 124.8 mmol)、メシルクロリド(5.79 ml, 74.9 mmol)を順次加え、10分間攪拌した。反応混合物に飽和塩化アンモニウム水溶液を加え、酢酸エチルで抽出した。有機層を飽和食塩水溶液で洗浄し、無水硫酸マグネシウムで乾燥後、減圧濃縮を行い、11-メタンスルホニルオキシ-1-ウンデセンを得た。11-メタンスルホニルオキシ-1-ウンデセン(3.00g, 12.08 mmol)のDMF(24.2 ML)溶液に0 ℃でアジ化ナトリウム(1.178 g, 18.12 mmol)を加え、50 ℃で3時間攪拌した。反応混合物に水を加え、ジエチルエーテルで抽出した。有機層を飽和食塩水溶液で洗浄し、無水硫酸マグネシウムで乾燥後、減圧濃縮を行った。残渣をシリカゲルクロマトグラフィー(ヘキサンに4.8％の酢酸エチルを溶かしたもの)により精製し、11-アジド-1-ウンデセンを得た(2.228 g, quant.)。11-アジド-1-ウンデセンの¹H NMR、¹³C NMRデータを以下に示す。 Triethylamine (17.4 ml, 124.8 mmol) and mesyl chloride (5.79 ml, 74.9 mmol) were sequentially added to a solution of 11-hydroxy-1-undecene (10.0 ml, 49.9 mmol) in THF (100 ml) at 0 ° C for 10 minutes. Stir. A saturated aqueous ammonium chloride solution was added to the reaction mixture, and the mixture was extracted with ethyl acetate. The organic layer was washed with a saturated aqueous sodium chloride solution, dried over anhydrous magnesium sulfate, and concentrated under reduced pressure to obtain 11-methanesulfonyloxy-1-undecene. Sodium azide (1.178 g, 18.12 mmol) was added to a DMF (24.2 ML) solution of 11-methanesulfonyloxy-1-undecene (3.00 g, 12.08 mmol) at 0 ° C., and the mixture was stirred at 50 ° C. for 3 hours. Water was added to the reaction mixture, and the mixture was extracted with diethyl ether. The organic layer was washed with a saturated aqueous sodium chloride solution, dried over anhydrous magnesium sulfate, and concentrated under reduced pressure. The residue was purified by silica gel chromatography (4.8% ethyl acetate dissolved in hexane) to give 11-azido-1-undecene (2.228 g, quant.). ¹ H NMR and ¹³ C NMR data of 11-azido-1-undecene are shown below.

¹H NMR (CDCl₃, 400 MHz) δ5.81 (tdd, J = 6.6, 10.3, 17.1 Hz, 1H), 4.99 (tdd, J = 1.5, 2.2, 17.1 Hz, 1H), 4.93 (tdd, J = 1.2, 2.2, 10.3 Hz, 1H), 3.25 (t, J = 7.1 Hz, 2H), 2.04 (m, 2H), 1.60 (m, 2H), 1.28 - 1.45(m, 14H)
¹³C NMR (CDCl₃, 100 MHz) δ139.2, 114.1, 51.5, 33.8, 29.4, 29.3, 29.11, 29.06, 28.9, 28.8, 26.7 ¹ H NMR (CDCl ₃ , 400 MHz) δ5.81 (tdd, J = 6.6, 10.3, 17.1 Hz, 1H), 4.99 (tdd, J = 1.5, 2.2, 17.1 Hz, 1H), 4.93 (tdd, J = 1.2, 2.2, 10.3 Hz, 1H), 3.25 (t, J = 7.1 Hz, 2H), 2.04 (m, 2H), 1.60 (m, 2H), 1.28-1.45 (m, 14H)
¹³ C NMR (CDCl ₃ , 100 MHz) δ139.2, 114.1, 51.5, 33.8, 29.4, 29.3, 29.11, 29.06, 28.9, 28.8, 26.7

（参考例２）

(Reference Example 2)

11-アジド-1-ウンデセン(11.49 g, 58.72 mmol)のTHF(147 ml)、水(14.7 ml)溶液に室温でトリフェニルホスフィン(23.98 g, 88.08 mmol)を加え、60 ℃で12時間攪拌した。反応混合物に水を加え、酢酸エチルで抽出した。有機層を飽和食塩水溶液で洗浄し、無水硫酸マグネシウムで乾燥後、減圧濃縮を行った。残渣を減圧蒸留(2 mmHg, 86 - 87 ℃)により精製し、11-アミノ-1-ウンデセンを得た(8.29 g, 83% yield)。11-アミノ-1-ウンデセンのIR、¹H NMR、¹³C NMRデータを以下に示す。 Triphenylphosphine (23.98 g, 88.08 mmol) was added to a solution of 11-azido-1-undecene (11.49 g, 58.72 mmol) in THF (147 ml) and water (14.7 ml) at room temperature, and the mixture was stirred at 60 ° C. for 12 hours. . Water was added to the reaction mixture, and the mixture was extracted with ethyl acetate. The organic layer was washed with a saturated aqueous sodium chloride solution, dried over anhydrous magnesium sulfate, and concentrated under reduced pressure. The residue was purified by distillation under reduced pressure (2 mmHg, 86-87 ° C.) to obtain 11-amino-1-undecene (8.29 g, 83% yield). IR, ¹ H NMR and ¹³ C NMR data of 11-amino-1-undecene are shown below.

IR (NaCl neat): 3296, 2924, 2854, 1642, 1462, 993, 909 cm^-1
¹H NMR (CDCl₃, 400 MHz) δ5.81 (tdd, J = 6.8, 10.3, 17.1 Hz, 1H), 4.99 (tdd, J = 1.5, 2.2, 17.1 Hz, 1H), 4.93 (tdd, J = 1.2, 2.2, 10.3 Hz, 1H), 2.68 (t, J = 6.8 Hz, 2H), 2.04 (m, 2H), 1.28 - 1.45(m, 14H)
¹³C NMR (CDCl₃, 100 MHz) δ139.2, 114.1, 42.3, 33.9, 33.8, 29.6, 29.5, 29.4, 29.1, 28.91, 26.87 IR (NaCl neat): 3296, 2924, 2854, 1642, 1462, 993, 909 cm ^-1
¹ H NMR (CDCl ₃ , 400 MHz) δ5.81 (tdd, J = 6.8, 10.3, 17.1 Hz, 1H), 4.99 (tdd, J = 1.5, 2.2, 17.1 Hz, 1H), 4.93 (tdd, J = 1.2, 2.2, 10.3 Hz, 1H), 2.68 (t, J = 6.8 Hz, 2H), 2.04 (m, 2H), 1.28-1.45 (m, 14H)
¹³ C NMR (CDCl ₃ , 100 MHz) δ139.2, 114.1, 42.3, 33.9, 33.8, 29.6, 29.5, 29.4, 29.1, 28.91, 26.87

（参考例３）

(Reference Example 3)

11-アミノ-1-ウンデセン(3.00 g, 17.72 mmol)のTHF溶液に0 ℃で4-クロロ-７-ニトロベンゾ［1,2,5］オキサジアゾール(3.536 g, 17.72 mmol)を加え、室温で1時間攪拌した。反応混合物に飽和塩化アンモニウム水溶液を加え、酢酸エチルで抽出した。有機層を飽和食塩水溶液で洗浄し、無水硫酸マグネシウムで乾燥後、減圧濃縮を行った。残渣をシリカゲルクロマトグラフィー(ヘキサンに９％の酢酸エチルを溶かしたもの)により精製し、11-[(7-ニトロベンゾ[1,2,5]オキサジアゾール-4-イル)アミノ]-1-ウンデセンを得た(3.56 g, 60% yield)。11-[(7-ニトロベンゾ[1,2,5]オキサジアゾール-4-イル)アミノ]-1-ウンデセンのIR、¹H NMR、¹³C NMRデータを以下に示す。 To a THF solution of 11-amino-1-undecene (3.00 g, 17.72 mmol) was added 4-chloro-7-nitrobenzo [1,2,5] oxadiazole (3.536 g, 17.72 mmol) at 0 ° C., and at room temperature. Stir for 1 hour. A saturated aqueous ammonium chloride solution was added to the reaction mixture, and the mixture was extracted with ethyl acetate. The organic layer was washed with a saturated aqueous sodium chloride solution, dried over anhydrous magnesium sulfate, and concentrated under reduced pressure. The residue was purified by silica gel chromatography (9% ethyl acetate in hexane) to give 11-[(7-nitrobenzo [1,2,5] oxadiazol-4-yl) amino] -1-undecene. (3.56 g, 60% yield). IR, ¹ H NMR, and ¹³ C NMR data of 11-[(7-nitrobenzo [1,2,5] oxadiazol-4-yl) amino] -1-undecene are shown below.

IR (KBr disk): 3418, 3324, 2924, 2855, 1622, 1561, 1300, 1258, 1111 cm^-1
¹H NMR (CDCl₃, 400 MHz) δ8.50 (d, J = 8.9 Hz, 1H), 6.19 (br m, 1H), 6.17 (d, J = 8.8 Hz, 1H), 5.81 (tdd, J = 6.8, 10.3, 17.1 Hz, 1H), 4.96 (tdd, J = 1.5, 2.2, 17.1 Hz, 1H), 4.93 (tdd, J = 1.2, 2.2, 10.3 Hz, 1H), 3.49 (m, 2H), 2.04 (m, 2H), 1.81 (tt, J = 7.3, 7.3 Hz, 2H), 1.30 - 1.51(m, 12H)
¹³C NMR (CDCl₃, 100 MHz) δ144.1, 144.0, 143.7, 138.9, 136.7, 123.0, 113.9, 98.5, 44.0, 33.6, 29.20, 29.15, 29.0, 28.8, 28.7, 28.3, 26.7 IR (KBr disk): 3418, 3324, 2924, 2855, 1622, 1561, 1300, 1258, 1111 cm ^-1
¹ H NMR (CDCl ₃ , 400 MHz) δ8.50 (d, J = 8.9 Hz, 1H), 6.19 (br m, 1H), 6.17 (d, J = 8.8 Hz, 1H), 5.81 (tdd, J = 6.8, 10.3, 17.1 Hz, 1H), 4.96 (tdd, J = 1.5, 2.2, 17.1 Hz, 1H), 4.93 (tdd, J = 1.2, 2.2, 10.3 Hz, 1H), 3.49 (m, 2H), 2.04 (m, 2H), 1.81 (tt, J = 7.3, 7.3 Hz, 2H), 1.30-1.51 (m, 12H)
¹³ C NMR (CDCl ₃ , 100 MHz) δ 144.1, 144.0, 143.7, 138.9, 136.7, 123.0, 113.9, 98.5, 44.0, 33.6, 29.20, 29.15, 29.0, 28.8, 28.7, 28.3, 26.7

（参考例４）

(Reference Example 4)

p-メトキシ安息香酸(0.94 g, 9.39 mmol)のトルエン(18.8 ml)溶液に室温でジフェニルリン酸アジド(DPPA)(2.23 ml, 10.33 mmol)、トリエチルアミン(1.44 ml, 10.33 mmol)を順次加え、10分間攪拌した後、(3R, 4S)-4,5-エポキシ-3-ヒドロキシ-1-ペンテン(1.50 g, 9,86 mmol)を加え、100 ℃に昇温し、1時間攪拌した。反応混合物に飽和塩化アンモニウム水溶液を加え、ジエチルエーテルで抽出した。有機層を飽和食塩水溶液で洗浄し、無水硫酸マグネシウムで乾燥後、減圧濃縮を行った。残渣をシリカゲルクロマトグラフィー(ヘキサンに17〜25％の酢酸エチルを溶かしたもの)により精製し、（(3R,4S)-4,5-エポキシ-1-ペンテン-3-イル）p-メトキシフェニルカルバメートを得た(1.829 g, 78% yield)。(3R,4S)-4,5-エポキシ-1-ペンテン-3-イル）p-メトキシフェニルカルバメートの比旋光度、IR、¹H NMR、¹³C NMRデータを以下に示す。 To a toluene (18.8 ml) solution of p-methoxybenzoic acid (0.94 g, 9.39 mmol), diphenylphosphoric acid azide (DPPA) (2.23 ml, 10.33 mmol) and triethylamine (1.44 ml, 10.33 mmol) were sequentially added at room temperature. After stirring for 3 minutes, (3R, 4S) -4,5-epoxy-3-hydroxy-1-pentene (1.50 g, 9,86 mmol) was added, and the temperature was raised to 100 ° C. and stirred for 1 hour. A saturated aqueous ammonium chloride solution was added to the reaction mixture, and the mixture was extracted with diethyl ether. The organic layer was washed with a saturated aqueous sodium chloride solution, dried over anhydrous magnesium sulfate, and concentrated under reduced pressure. The residue was purified by silica gel chromatography (17-25% ethyl acetate in hexane) and ((3R, 4S) -4,5-epoxy-1-penten-3-yl) p-methoxyphenylcarbamate (1.829 g, 78% yield). Specific rotation, IR, ¹ H NMR and ¹³ C NMR data of (3R, 4S) -4,5-epoxy-1-penten-3-yl) p-methoxyphenylcarbamate are shown below.

[α]D^23.0 +23.9 (c = 0.82, CHCl₃)
IR (KBr disk): 3333, 2955, 1701, 1597, 1526, 1231, 1051 cm^-1
¹H NMR (CDCl₃, 400 MHz) δ7.28 (md, J = 8.1 Hz, 2H), 6.85 (md, J = 9.0 Hz, 2H), 6.58 (brm, 1H), 5.84 (ddd, J = 6.8, 10.5, 17.3 Hz, 1H), 5.45 (md, J = 17.3 Hz, 1H), 5.35 (md, J = 10.5 Hz, 1H), 5.29 (m, 1H), 3.18 (ddd, J = 2.7, 3.9, 3.9 Hz, 1H), 2.81 (dd, J = 4.2, 4.9 Hz, 1H), 2.72 (dd, J = 2.7, 4.9 Hz, 1H)
¹³C NMR (CDCl₃, 100 MHz) δ156.2, 142.8, 131.7, 129.3, 120.7, 120.0, 114.3, 74.5, 44.5, 52.1, 44.8. [α] D ^23.0 +23.9 (c = 0.82, CHCl ₃ )
IR (KBr disk): 3333, 2955, 1701, 1597, 1526, 1231, 1051 cm ^-1
¹ H NMR (CDCl ₃ , 400 MHz) δ7.28 (md, J = 8.1 Hz, 2H), 6.85 (md, J = 9.0 Hz, 2H), 6.58 (brm, 1H), 5.84 (ddd, J = 6.8 , 10.5, 17.3 Hz, 1H), 5.45 (md, J = 17.3 Hz, 1H), 5.35 (md, J = 10.5 Hz, 1H), 5.29 (m, 1H), 3.18 (ddd, J = 2.7, 3.9, 3.9 Hz, 1H), 2.81 (dd, J = 4.2, 4.9 Hz, 1H), 2.72 (dd, J = 2.7, 4.9 Hz, 1H)
¹³ C NMR (CDCl ₃ , 100 MHz) δ156.2, 142.8, 131.7, 129.3, 120.7, 120.0, 114.3, 74.5, 44.5, 52.1, 44.8.

（参考例５）

(Reference Example 5)

（(3R,4S)-4,5-エポキシ-1-ペンテン-3-イル）p-メトキシフェニルカルバメート (13.88 g, 55.69 mmol)のTHF(111 ml)、DMF(11 ml)溶液に0 ℃で水素化ナトリウム(0.267 g, 11.14 mmol)を加え、45分間攪拌した。反応混合物に飽和塩化アンモニウム水溶液を加え、ジエチルエーテルで抽出した。有機層を飽和食塩水溶液で洗浄し、無水硫酸マグネシウムで乾燥後、減圧濃縮を行った。残渣をシリカゲルクロマトグラフィー(ヘキサンに33〜67％の酢酸エチルを溶かしたもの)により精製し、(4R,5R)-4-ヒドロキシメチル-3-p-メトキシフェニル-5-ビニルオキサゾリジン-2-オンを得た(13.68g, 99% yield)。(4R,5R)-4-ヒドロキシメチル-3-p-メトキシフェニル-5-ビニルオキサゾリジン-2-オンの比旋光度、IR、¹H NMR、¹³C NMRデータを以下に示す。 ((3R, 4S) -4,5-epoxy-1-penten-3-yl) p-methoxyphenylcarbamate (13.88 g, 55.69 mmol) in THF (111 ml) and DMF (11 ml) at 0 ° C Sodium hydride (0.267 g, 11.14 mmol) was added and stirred for 45 minutes. A saturated aqueous ammonium chloride solution was added to the reaction mixture, and the mixture was extracted with diethyl ether. The organic layer was washed with a saturated aqueous sodium chloride solution, dried over anhydrous magnesium sulfate, and concentrated under reduced pressure. The residue was purified by silica gel chromatography (33-67% ethyl acetate in hexane) and (4R, 5R) -4-hydroxymethyl-3-p-methoxyphenyl-5-vinyloxazolidine-2-one (13.68g, 99% yield). Specific rotation, IR, ¹ H NMR, and ¹³ C NMR data of (4R, 5R) -4-hydroxymethyl-3-p-methoxyphenyl-5-vinyloxazolidine-2-one are shown below.

[α]D^23.5 +5.8 (c = 0.82, CHCl₃)
IR (KBr disk): 3319, 2909, 1721, 1611, 1518, 1426, 1254, 1155, 1030 cm^-1
¹H NMR (CDCl₃, 400 MHz)δ7.30 (md, J = 8.8 Hz, 2H), 6.93 (md, J = 9.0 Hz, 2H), 6.01 (ddd, J = 6.3, 10.5, 17.3 Hz, 1H), 5.54 (ddd, J = 0.98, 0.98, 17.3 Hz, 1H), 5.39 (ddd, J = 0.98, 0.98, 10.5 Hz, 1H), 5.01 (m, 1H), 4.02 (ddd, J = 2.7, 4.1, 6.1 Hz, 1H), 3.81 (m, 1H), 3.81 (s, 3H), 3.67 (ddd, J = 2.7, 6.6, 12.0 Hz, 1H)
¹³C NMR (CDCl₃, 100 MHz) δ157.9, 134.3, 128.9, 125.2, 125.1, 118.9, 114.6, 76.3, 64.0, 59.6, 55.5 [α] D ^23.5 +5.8 (c = 0.82, CHCl ₃ )
IR (KBr disk): 3319, 2909, 1721, 1611, 1518, 1426, 1254, 1155, 1030 cm ^-1
¹ H NMR (CDCl ₃ , 400 MHz) δ 7.30 (md, J = 8.8 Hz, 2H), 6.93 (md, J = 9.0 Hz, 2H), 6.01 (ddd, J = 6.3, 10.5, 17.3 Hz, 1H ), 5.54 (ddd, J = 0.98, 0.98, 17.3 Hz, 1H), 5.39 (ddd, J = 0.98, 0.98, 10.5 Hz, 1H), 5.01 (m, 1H), 4.02 (ddd, J = 2.7, 4.1 , 6.1 Hz, 1H), 3.81 (m, 1H), 3.81 (s, 3H), 3.67 (ddd, J = 2.7, 6.6, 12.0 Hz, 1H)
¹³ C NMR (CDCl ₃ , 100 MHz) δ157.9, 134.3, 128.9, 125.2, 125.1, 118.9, 114.6, 76.3, 64.0, 59.6, 55.5

（参考例６）

(Reference Example 6)

(4R,5R)-4-ヒドロキシメチル-3-p-メトキシフェニル-5-ビニルオキサゾリジン-2-オン (0.500 g, 2.01 mmol)の塩化メチレン(20.1 ml)溶液に-78 ℃で2,6-ルチジン(0.304 ml, 2.61 ml)、トリフルオロメタンスルホン酸無水物(0.439 ml, 2.61 mmol)を順次加え、10分間攪拌した。反応混合物に飽和炭酸水素ナトリウム水溶液を加え、クロロホルムで抽出した。有機層を飽和食塩水溶液で洗浄し、無水硫酸マグネシウムで乾燥後、減圧濃縮を行った。残渣をシリカゲルクロマトグラフィー(ヘキサンに17〜50％の酢酸エチルを溶かしたもの)により精製し、(4S,5R)-4-トリフルオロメタンスルホニルオキシメチル-3-p-メトキシフェニル-5-ビニルオキサゾリジン-2-オンを得た(0.743 g, 97% yield)。(4S,5R)-4-トリフルオロメタンスルホニルオキシメチル-3-p-メトキシフェニル-5-ビニルオキサゾリジン-2-オンのIR、¹H NMR、¹³C NMRデータを以下に示す。 (4R, 5R) -4-Hydroxymethyl-3-p-methoxyphenyl-5-vinyloxazolidin-2-one (0.500 g, 2.01 mmol) in methylene chloride (20.1 ml) at −78 ° C. Lutidine (0.304 ml, 2.61 ml) and trifluoromethanesulfonic anhydride (0.439 ml, 2.61 mmol) were sequentially added and stirred for 10 minutes. A saturated aqueous sodium hydrogen carbonate solution was added to the reaction mixture, and the mixture was extracted with chloroform. The organic layer was washed with a saturated aqueous sodium chloride solution, dried over anhydrous magnesium sulfate, and concentrated under reduced pressure. The residue was purified by silica gel chromatography (17-50% ethyl acetate in hexane) and (4S, 5R) -4-trifluoromethanesulfonyloxymethyl-3-p-methoxyphenyl-5-vinyloxazolidine- 2-one was obtained (0.743 g, 97% yield). IR, ¹ H NMR, and ¹³ C NMR data of (4S, 5R) -4-trifluoromethanesulfonyloxymethyl-3-p-methoxyphenyl-5-vinyloxazolidin-2-one are shown below.

IR (KBr disk): 3449, 1732, 1516, 1410, 1248, 1221, 1150 cm^-1
¹H NMR (CDCl₃, 400 MHz) δ7.26 (md, J = 9.0 Hz, 2H), 6.95 (md, J = 9.0 Hz, 2H), 6.01 (ddd, J = 6.6, 10.5, 17.1 Hz, 1H), 5.59 (ddd, J = 0.73, 1.2, 17.1 Hz, 1H), 5.48 (ddd, J = 0.73, 0.98, 10.5 Hz, 1H), 4.92 (dddd, J = 0.98, 1.2, 5.6, 6.3 Hz, 1H), 4.54 (dd, J = 4.6, 11.0 Hz, 1H), 4.50 (dd, J = 3.2, 11.0 Hz, 1H), 4.26 (ddd, J = 3.2, 4.6, 5.4 Hz, 1H), 3.82 (s, 3H)
¹³C NMR (CDCl₃, 100 MHz) δ158.6, 154.9, 132.9, 127.6, 125.4, 120.5, 75.7, 71.5, 61.3, 55.5 IR (KBr disk): 3449, 1732, 1516, 1410, 1248, 1221, 1150 cm ^-1
¹ H NMR (CDCl ₃ , 400 MHz) δ7.26 (md, J = 9.0 Hz, 2H), 6.95 (md, J = 9.0 Hz, 2H), 6.01 (ddd, J = 6.6, 10.5, 17.1 Hz, 1H ), 5.59 (ddd, J = 0.73, 1.2, 17.1 Hz, 1H), 5.48 (ddd, J = 0.73, 0.98, 10.5 Hz, 1H), 4.92 (dddd, J = 0.98, 1.2, 5.6, 6.3 Hz, 1H ), 4.54 (dd, J = 4.6, 11.0 Hz, 1H), 4.50 (dd, J = 3.2, 11.0 Hz, 1H), 4.26 (ddd, J = 3.2, 4.6, 5.4 Hz, 1H), 3.82 (s, 3H)
¹³ C NMR (CDCl ₃ , 100 MHz) δ158.6, 154.9, 132.9, 127.6, 125.4, 120.5, 75.7, 71.5, 61.3, 55.5

（参考例７）

(Reference Example 7)

メチルホスホン酸ジメチル(3.19 ml, 29.41 mmol)のTHF(46 ml)溶液に-78 ℃でn-ブチルリチウムの1.6Nヘキサン溶液(17.4 ml, 28.86 mmol)を滴下し、同温で30分間攪拌した後、同温で(4S,5R)-4-トリフルオロメタンスルホニルオキシメチル-3-p-メトキシフェニル-5-ビニルオキサゾリジン-2-オン (5.903 g, 15.48 mmol)のTHF(46 ml)溶液を滴下し、10分間攪拌した。反応混合物に反応混合物に飽和塩化アンモニウム水溶液を加え、酢酸エチルで抽出した。有機層を飽和食塩水溶液で洗浄し、無水硫酸マグネシウムで乾燥後、減圧濃縮を行った。残渣をシリカゲルクロマトグラフィー（クロロホルムに0〜４%のメタノールを溶かしたもの)により精製し、[2-((4R,5R)-3-p-メトキシフェニル-2-オキソ-5-ビニルオキサゾリジン-4-イル)エチル] ホスホン酸ジメチルエステルを得た(3.957 g, 72% yield)。[2-((4R,5R)-3-p-メトキシフェニル-2-オキソ-5-ビニルオキサゾリジン-4-イル)エチル] ホスホン酸ジメチルエステルの比旋光度、IR、¹H NMR、¹³C NMRデータを以下に示す。 To a solution of dimethyl methylphosphonate (3.19 ml, 29.41 mmol) in THF (46 ml) was added dropwise a 1.6N hexane solution of n-butyllithium (17.4 ml, 28.86 mmol) at -78 ° C and stirred at the same temperature for 30 minutes. (4S, 5R) -4-trifluoromethanesulfonyloxymethyl-3-p-methoxyphenyl-5-vinyloxazolidine-2-one (5.903 g, 15.48 mmol) in THF (46 ml) was added dropwise at the same temperature. And stirred for 10 minutes. A saturated aqueous ammonium chloride solution was added to the reaction mixture, and the mixture was extracted with ethyl acetate. The organic layer was washed with a saturated aqueous sodium chloride solution, dried over anhydrous magnesium sulfate, and concentrated under reduced pressure. The residue was purified by silica gel chromatography (0-4% methanol in chloroform) and [2-((4R, 5R) -3-p-methoxyphenyl-2-oxo-5-vinyloxazolidine-4 -Yl) ethyl] phosphonic acid dimethyl ester was obtained (3.957 g, 72% yield). [2-((4R, 5R) -3-p-methoxyphenyl-2-oxo-5-vinyloxazolidin-4-yl) ethyl] Specific rotation of phosphonic acid dimethyl ester, IR, ¹ H NMR, ¹³ C NMR The data is shown below.

[α]D^23.5 -4.1 (c = 1.06, CHCl₃)
IR (KBr disk): 3015, 2955, 1748, 1516, 1429, 1408, 1242, 993, 820 cm^-1
¹H NMR (CDCl₃, 400 MHz) δ7.29 (md, J = 9.0 Hz, 2H), 6.93 (md, J = 9.0 Hz, 2H), 5.98 (ddd, J = 6.6, 10.5, 17.1 Hz, 1H), 5.53 (ddd, J = 0.73, 1.2, 17.1 Hz, 1H), 5.41 (ddd, J = 0.73, 0.98, 10.5 Hz, 1H), 4.65 (dddd, J = 0.98, 1.2, 5.6, 6.6 Hz, 1H), 4.11 (ddd, J = 3.9, 5.6, 6.6 Hz, 1H), 3.82 (s, 3H), 3.69 (d, J = 10.7 Hz, 3H), 3.67 (d, J = 11.0 Hz, 3H), 1.91 - 2.02 (m, 2H), 1.64 - 1.73 (m, 2H)
¹³C NMR (CDCl₃, 100 MHz) δ157.6, 155.2, 134.1, 128.8, 124.6, 119.4, 114.5, 78.2, 61.7 (d, Jc-p = 17.4 Hz), 55.3, 52.4 (d, Jc-p = 6.6 Hz), 52.3 (d, Jc-p = 6.6 Hz), 24.3 (d, Jc-p = 4.1 Hz), 19.0 (d, Jc-p = 143.9 Hz) [α] D ^23.5 -4.1 (c = 1.06, CHCl ₃ )
IR (KBr disk): 3015, 2955, 1748, 1516, 1429, 1408, 1242, 993, 820 cm ^-1
¹ H NMR (CDCl ₃ , 400 MHz) δ7.29 (md, J = 9.0 Hz, 2H), 6.93 (md, J = 9.0 Hz, 2H), 5.98 (ddd, J = 6.6, 10.5, 17.1 Hz, 1H ), 5.53 (ddd, J = 0.73, 1.2, 17.1 Hz, 1H), 5.41 (ddd, J = 0.73, 0.98, 10.5 Hz, 1H), 4.65 (dddd, J = 0.98, 1.2, 5.6, 6.6 Hz, 1H ), 4.11 (ddd, J = 3.9, 5.6, 6.6 Hz, 1H), 3.82 (s, 3H), 3.69 (d, J = 10.7 Hz, 3H), 3.67 (d, J = 11.0 Hz, 3H), 1.91 -2.02 (m, 2H), 1.64-1.73 (m, 2H)
¹³ C NMR (CDCl ₃ , 100 MHz) δ157.6, 155.2, 134.1, 128.8, 124.6, 119.4, 114.5, 78.2, 61.7 (d, Jc-p = 17.4 Hz), 55.3, 52.4 (d, Jc-p = 6.6 Hz), 52.3 (d, Jc-p = 6.6 Hz), 24.3 (d, Jc-p = 4.1 Hz), 19.0 (d, Jc-p = 143.9 Hz)

（参考例８）

(Reference Example 8)

[2-((4R,5R)-3-p-メトキシフェニル-2-オキソ-5-ビニルオキサゾリジン-4-イル)エチル] ホスホン酸ジメチルエステル (0.100 g, 0.281 mmol)のアセトニトリル(4.3 ml), 水(1.4 ml)に、-15 ℃で硝酸二アンモニウムセリウム(IV)（CAN）(0.617 g, 1.126 mmol)を加え、7分間攪拌した。反応混合物に飽和食塩水溶液を加え、酢酸エチルで抽出した。有機層を飽和食塩水溶液で洗浄し、無水硫酸マグネシウムで乾燥後、減圧濃縮を行った。残渣をシリカゲルクロマトグラフィー(クロロホルムに0〜９%のメタノールを溶かしたもの)により粗精製し、[2-((4R,5R)-2-オキソ-5-ビニルオキサゾリジン-4-イル)エチル] ホスホン酸ジメチルエステルを得た(0.059 g, 84% yield)。[2-((4R,5R)-2-オキソ-5-ビニルオキサゾリジン-4-イル)エチル] ホスホン酸ジメチルエステルの¹H NMR、¹³C NMRデータを以下に示す。 [2-((4R, 5R) -3-p-methoxyphenyl-2-oxo-5-vinyloxazolidin-4-yl) ethyl] phosphonic acid dimethyl ester (0.100 g, 0.281 mmol) in acetonitrile (4.3 ml), To water (1.4 ml), diammonium cerium (IV) nitrate (CAN) (0.617 g, 1.126 mmol) was added at −15 ° C. and stirred for 7 minutes. A saturated aqueous sodium chloride solution was added to the reaction mixture, and the mixture was extracted with ethyl acetate. The organic layer was washed with a saturated aqueous sodium chloride solution, dried over anhydrous magnesium sulfate, and concentrated under reduced pressure. The residue was roughly purified by silica gel chromatography (0-9% methanol in chloroform) to give [2-((4R, 5R) -2-oxo-5-vinyloxazolidine-4-yl) ethyl] phosphone. Acid dimethyl ester was obtained (0.059 g, 84% yield). [2-((4R, 5R) -2-oxo-5-vinyloxazolidin-4-yl) ethyl] ¹ H NMR and ¹³ C NMR data of phosphonic acid dimethyl ester are shown below.

¹H NMR (CDCl₃, 400 MHz) δ6.58 (br d, 5.1 Hz, 1H), 5.90 (ddd, J = 6.8, 10.3, 17.1 Hz, 1H), 5.44 (ddd, J = 0.73, 0.98, 17.1 Hz, 1H), 5.35 (md, J = 10.3 Hz, 1H), 4.55 (mdd, J = 6.6, 6.6 Hz, 1H), 3.78 (d, J = 11.0 Hz, 3H), 3.77 (d, J = 10.8 Hz, 3H), 3.63 (m, 1H), 1.81 - 1.97 (m, 4H)
¹³C NMR (CDCl₃, 100 MHz) δ158.6, 133.8, 119.4, 82.4, 58.22 (d, Jc-p = 14.1 Hz), 52.7 (d, Jc-p = 6.6 Hz), 52.6 (d, Jc-p = 6.6 Hz), 27.5 (d, Jc-p = 5.0 Hz), 21.0 (d, Jc-p = 143.9 Hz) ¹ H NMR (CDCl ₃ , 400 MHz) δ6.58 (br d, 5.1 Hz, 1H), 5.90 (ddd, J = 6.8, 10.3, 17.1 Hz, 1H), 5.44 (ddd, J = 0.73, 0.98, 17.1 Hz, 1H), 5.35 (md, J = 10.3 Hz, 1H), 4.55 (mdd, J = 6.6, 6.6 Hz, 1H), 3.78 (d, J = 11.0 Hz, 3H), 3.77 (d, J = 10.8 Hz, 3H), 3.63 (m, 1H), 1.81-1.97 (m, 4H)
¹³ C NMR (CDCl ₃ , 100 MHz) δ158.6, 133.8, 119.4, 82.4, 58.22 (d, Jc-p = 14.1 Hz), 52.7 (d, Jc-p = 6.6 Hz), 52.6 (d, Jc- p = 6.6 Hz), 27.5 (d, Jc-p = 5.0 Hz), 21.0 (d, Jc-p = 143.9 Hz)

（参考例９）

(Reference Example 9)

[2-((4R,5R)-2-オキソ-5-ビニルオキサゾリジン-4-イル)エチル] ホスホン酸ジメチルエステル (0.423 g, 1.69 mmol)の塩化メチレン(17.0 ml)溶液に0 ℃でジメチルアミノピリジン(DMAP)(0.104 g, 0.849 mmol)、トリエチルアミン(0.473 ml, 3.39 mmol)、ジ-t-ブチルジカーボネート（Boc₂O）(0.443 g, 2.546 mmol)を順次加え、30分間攪拌した。反応混合物に飽和塩化アンモニウム水溶液を加え、クロロホルムで抽出した。有機層を飽和食塩水溶液で洗浄し、無水硫酸マグネシウムで乾燥後、減圧濃縮を行った。残渣をシリカゲルクロマトグラフィー(クロロホルムに0〜２%のメタノールを溶かしたもの）により精製し、[2-((4R,5R)-3-t-ブトキシカルボニル-2-オキソ-5-ビニルオキサゾリジン-4-イル)エチル] ホスホン酸ジメチルエステルを得た(0.363 g, 61% yield)。[2-((4R,5R)-3-t-ブトキシカルボニル-2-オキソ-5-ビニルオキサゾリジン-4-イル)エチル] ホスホン酸ジメチルエステルのIR、¹H NMR、¹³C NMRデータを以下に示す。 [2-((4R, 5R) -2-oxo-5-vinyloxazolidine-4-yl) ethyl] Phosphonic acid dimethyl ester (0.423 g, 1.69 mmol) in methylene chloride (17.0 ml) at 0 ° C with dimethylamino Pyridine (DMAP) (0.104 g, 0.849 mmol), triethylamine (0.473 ml, 3.39 mmol) and di-t-butyl dicarbonate (Boc ₂ O) (0.443 g, 2.546 mmol) were sequentially added and stirred for 30 minutes. A saturated aqueous ammonium chloride solution was added to the reaction mixture, and the mixture was extracted with chloroform. The organic layer was washed with a saturated aqueous sodium chloride solution, dried over anhydrous magnesium sulfate, and concentrated under reduced pressure. The residue was purified by silica gel chromatography (0 to 2% methanol dissolved in chloroform) and [2-((4R, 5R) -3-t-butoxycarbonyl-2-oxo-5-vinyloxazolidine-4 -Yl) ethyl] phosphonic acid dimethyl ester was obtained (0.363 g, 61% yield). [2-((4R, 5R) -3-t-butoxycarbonyl-2-oxo-5-vinyloxazolidin-4-yl) ethyl] IR, ¹ H NMR and ¹³ C NMR data of phosphonic acid dimethyl ester are shown below. Show.

IR (NaCl neat): 2982, 1817, 1725, 1372, 1327, 1256, 1159cm^-1
¹H NMR (CDCl₃, 400 MHz) δ5.87 (ddd, J = 6.1, 10.5, 17.1 Hz, 1H), 5.46 (ddd, J = 0.73, 1.2, 17.1 Hz, 1H), 5.36 (md, J = 10.5 Hz, 1H), 4.56 (m, 1H), 4.05 (m, 1H), 3.76 (d, J = 11.0 Hz, 6H), 2.06 - 2.17 (m, 2H), 1.71 - 1.81 (m, 2H), 1.55 (s, 9H)
¹³C NMR (CDCl₃, 100 MHz) δ151.3, 149.2, 133.5, 119.2, 84.4, 77.5, 59.8 (d, Jc-p = 18.2 Hz), 52.6 (d, Jc-p = 6.6 Hz), 52.6 (d, Jc-p = 5.8 Hz), 27.9, 25.7 (d, Jc-p = 4.1 Hz), 19.7 (d, Jc-p = 144.8 Hz) IR (NaCl neat): 2982, 1817, 1725, 1372, 1327, 1256, 1159cm ^-1
¹ H NMR (CDCl ₃ , 400 MHz) δ5.87 (ddd, J = 6.1, 10.5, 17.1 Hz, 1H), 5.46 (ddd, J = 0.73, 1.2, 17.1 Hz, 1H), 5.36 (md, J = 10.5 Hz, 1H), 4.56 (m, 1H), 4.05 (m, 1H), 3.76 (d, J = 11.0 Hz, 6H), 2.06-2.17 (m, 2H), 1.71-1.81 (m, 2H), 1.55 (s, 9H)
¹³ C NMR (CDCl ₃ , 100 MHz) δ151.3, 149.2, 133.5, 119.2, 84.4, 77.5, 59.8 (d, Jc-p = 18.2 Hz), 52.6 (d, Jc-p = 6.6 Hz), 52.6 ( d, Jc-p = 5.8 Hz), 27.9, 25.7 (d, Jc-p = 4.1 Hz), 19.7 (d, Jc-p = 144.8 Hz)

（参考例１０）

(Reference Example 10)

[(1R,2R)-1-(tert-ブチルジメチルシラニルオキシメチル)-2-ヒドロキシブタ-3-エニル]-カルバミン酸tert-ブチルエステル(7.304 g, 22.03 mmol)のTHF(110 ml)溶液に０℃で水素化ナトリウム(793 mg, 33.05 mmol)を加え、５０℃に昇温し２時間撹拌した。反応混合物に飽和塩化アンモニウム水溶液を加え中和した後、酢酸エチルで抽出した。有機層を飽和食塩水で洗浄し、無水硫酸マグネシウムで乾燥後、減圧濃縮を行った。残渣をシリカゲルクロマトグラフィー（ヘキサンに13〜50％の酢酸エチルを溶かしたもの）により精製し、(4R,5R)-4-(tert-ブチルジメチルシラニルオキシメチル)-5-ビニルオキサゾリジン-2-オン(5.264 g, 93% yield)を得た。(4R,5R)-4-(tert-ブチルジメチルシラニルオキシメチル)-5-ビニルオキサゾリジン-2-オンの¹H NMR、¹³C NMRデータを以下に示す。 [(1R, 2R) -1- (tert-butyldimethylsilanyloxymethyl) -2-hydroxybut-3-enyl] -carbamic acid tert-butyl ester (7.304 g, 22.03 mmol) in THF (110 ml) To the mixture was added sodium hydride (793 mg, 33.05 mmol) at 0 ° C., and the mixture was heated to 50 ° C. and stirred for 2 hours. The reaction mixture was neutralized with a saturated aqueous ammonium chloride solution, and extracted with ethyl acetate. The organic layer was washed with saturated brine, dried over anhydrous magnesium sulfate, and concentrated under reduced pressure. The residue was purified by silica gel chromatography (13-50% ethyl acetate in hexane) and (4R, 5R) -4- (tert-butyldimethylsilanyloxymethyl) -5-vinyloxazolidine-2- On (5.264 g, 93% yield) was obtained. ¹ H NMR and ¹³ C NMR data of (4R, 5R) -4- (tert-butyldimethylsilanyloxymethyl) -5-vinyloxazolidine-2- ^one are shown below.

¹H NMR (CDCl₃, 400 MHz), δ5.90 (ddd, J = 6.8, 10.6, 17.2 Hz, 1H), 5.49 (ddd, J = 1.0, 2.1, 17.2 Hz, 1H), 5.37 (ddd, J = 1.2, 2.0, 10.6 Hz, 1H), 5.09 (m, 1H), 3.89 (ddd, J = 4.3, 7.5, 8.1 Hz, 1H), 3.59 (dd, J = 4.3, 10.3 Hz, 1H), 3.54 (dd, J = 7.4, 10.3 Hz, 1H), 0.89 (s, 9H), 0.061 (s, 3H), 0.057 (s, 3H).
¹³C NMR (CDCl₃, 100 MHz), δ158.7, 130.3, 120.2, 78.8, 62.4, 56.9, 25.7, 18.1, -5.5. ¹ H NMR (CDCl ₃ , 400 MHz), δ5.90 (ddd, J = 6.8, 10.6, 17.2 Hz, 1H), 5.49 (ddd, J = 1.0, 2.1, 17.2 Hz, 1H), 5.37 (ddd, J = 1.2, 2.0, 10.6 Hz, 1H), 5.09 (m, 1H), 3.89 (ddd, J = 4.3, 7.5, 8.1 Hz, 1H), 3.59 (dd, J = 4.3, 10.3 Hz, 1H), 3.54 ( dd, J = 7.4, 10.3 Hz, 1H), 0.89 (s, 9H), 0.061 (s, 3H), 0.057 (s, 3H).
¹³ C NMR (CDCl ₃ , 100 MHz), δ158.7, 130.3, 120.2, 78.8, 62.4, 56.9, 25.7, 18.1, -5.5.

（参考例１１）

(Reference Example 11)

(4R,5R)-4-(tert-ブチルジメチルシラニルオキシメチル)-5-ビニルオキサゾリジン-2-オン (7.365 g, 28.65 mmol)のTHF(150 ml)溶液に０℃で水素化ナトリウム(1.719 g, 42.98 mmol)を加え30分撹拌した後4-メトキシベンジルクロリド(5.83 ml, 43.0 mmol)、テトラブチルアンモニウムヨージド(21.267 g, 57.306 mmol)を順次加え、室温で１３時間撹拌した。反応混合物を飽和塩化アンモニウム水溶液で中和した後、酢酸エチルで抽出した。有機層を飽和食塩水で洗浄し、無水硫酸マグネシウムで乾燥後、減圧濃縮を行った。残渣をシリカゲルクロマトグラフィー（ヘキサンに７〜25％の酢酸エチルを溶かしたもの）により精製し、(4R,5R)-4-(tert-ブチルジメチルシラニルオキシメチル)-3-(4-メトキシベンジル)-5-ビニルオキサゾリジン-2-オン(10.882 g, quant.)を得た。(4R,5R)-4-(tert-ブチルジメチルシラニルオキシメチル)-3-(4-メトキシベンジル)-5-ビニルオキサゾリジン-2-オンの¹H NMR、¹³C NMRデータを以下に示す。 Sodium hydride (1.719) was added to a solution of (4R, 5R) -4- (tert-butyldimethylsilanyloxymethyl) -5-vinyloxazolidine-2-one (7.365 g, 28.65 mmol) in THF (150 ml) at 0 ° C. g, 42.98 mmol) was added and the mixture was stirred for 30 minutes, 4-methoxybenzyl chloride (5.83 ml, 43.0 mmol) and tetrabutylammonium iodide (21.267 g, 57.306 mmol) were successively added, and the mixture was stirred at room temperature for 13 hours. The reaction mixture was neutralized with saturated aqueous ammonium chloride solution and extracted with ethyl acetate. The organic layer was washed with saturated brine, dried over anhydrous magnesium sulfate, and concentrated under reduced pressure. The residue was purified by silica gel chromatography (7-25% ethyl acetate in hexane) and (4R, 5R) -4- (tert-butyldimethylsilanyloxymethyl) -3- (4-methoxybenzyl). ) -5-vinyloxazolidine-2-one (10.882 g, quant.) Was obtained. ¹ H NMR and ¹³ C NMR data of (4R, 5R) -4- (tert-butyldimethylsilanyloxymethyl) -3- (4-methoxybenzyl) -5-vinyloxazolidine-2- ^one are shown below.

¹H NMR (CDCl₃, 400 MHz), δ7.23 (md, J = 8.7 Hz, 2H), 6.87 (md, J = 8.7 Hz, 2H), 5.99 (ddd, J = 7.1, 10.6, 17.4 Hz, 1H), 5.41 (ddd, J = 1.1, 1.2, 17.4 Hz, 1H), 5.34 (ddd, J = 0.9, 1.1, 10.5 Hz, 1H), 4.89 (m, 1H), 4.89 (m, 1H), 4.85 (d, J = 15.1 Hz, 1H), 3.99 (s, 3H), 3.68 (m, 1H), 3.55 - 3.62 (m, 2H), 0.90 (s, 9H), 0.070 (s, 3H), 0.053 (s, 3H).
¹³C NMR (CDCl₃, 100 MHz), δ159.3, 158.1, 131.5, 129.4, 128.3, 120.3, 114.1, 77.6, 59.2, 58.5, 55.3, 45.9, 25.7, 18.0, -5.7. ¹ H NMR (CDCl ₃ , 400 MHz), δ7.23 (md, J = 8.7 Hz, 2H), 6.87 (md, J = 8.7 Hz, 2H), 5.99 (ddd, J = 7.1, 10.6, 17.4 Hz, 1H), 5.41 (ddd, J = 1.1, 1.2, 17.4 Hz, 1H), 5.34 (ddd, J = 0.9, 1.1, 10.5 Hz, 1H), 4.89 (m, 1H), 4.89 (m, 1H), 4.85 (d, J = 15.1 Hz, 1H), 3.99 (s, 3H), 3.68 (m, 1H), 3.55-3.62 (m, 2H), 0.90 (s, 9H), 0.070 (s, 3H), 0.053 ( s, 3H).
¹³ C NMR (CDCl ₃ , 100 MHz), δ159.3, 158.1, 131.5, 129.4, 128.3, 120.3, 114.1, 77.6, 59.2, 58.5, 55.3, 45.9, 25.7, 18.0, -5.7.

（参考例１２）

(Reference Example 12)

(4R,5R)-4-(tert-ブチルジメチルシラニルオキシメチル)-3-(4-メトキシベンジル)-5-ビニルオキサゾリジン-2-オン (10.731 g, 28.426 mmol)のMeOH(150 ml)溶液に0℃で2N-HCl(57 ml)を加え、室温で１２時間撹拌した。反応混合物に飽和炭酸水素ナトリウム水溶液で中和した後酢酸エチルで抽出した。有機層を飽和食塩水で洗浄し、無水硫酸マグネシウムで乾燥後、減圧濃縮を行った。残渣をシリカゲルクロマトグラフィー（ヘキサンに33〜67％の酢酸エチルを溶かしたもの）により精製し、(4S,5R)-4-ヒドロキシメチル-3-(4-メトキシベンジル)-5-ビニルオキサゾリジン-2-オン(7.447 g, 99% yield)を得た。(4S,5R)-4-ヒドロキシメチル-3-(4-メトキシベンジル)-5-ビニルオキサゾリジン-2-オンの¹H NMR、¹³C NMRデータを以下に示す。 (4R, 5R) -4- (tert-butyldimethylsilanyloxymethyl) -3- (4-methoxybenzyl) -5-vinyloxazolidine-2-one (10.731 g, 28.426 mmol) in MeOH (150 ml) To the mixture was added 2N-HCl (57 ml) at 0 ° C., and the mixture was stirred at room temperature for 12 hours. The reaction mixture was neutralized with saturated aqueous sodium hydrogen carbonate solution and extracted with ethyl acetate. The organic layer was washed with saturated brine, dried over anhydrous magnesium sulfate, and concentrated under reduced pressure. The residue was purified by silica gel chromatography (33-67% ethyl acetate in hexane) to give (4S, 5R) -4-hydroxymethyl-3- (4-methoxybenzyl) -5-vinyloxazolidine-2 -Obtained ON (7.447 g, 99% yield). ¹ H NMR and ¹³ C NMR data of (4S, 5R) -4-hydroxymethyl-3- (4-methoxybenzyl) -5-vinyloxazolidin-2- ^one are shown below.

¹H NMR (CDCl₃, 400 MHz), δ7.26 (md, J = 8.7 Hz, 2H), 6.88 (md, J = 8.7 Hz, 2H), 6.05 (ddd, J = 6.9, 10.5, 17.2 Hz, 1H), 5.50 (md, J = 17.2 Hz, 1H), 5.40 (md, J = 10.6 Hz, 1H), 4.94 (mdd, J = 7.1, 8.0 Hz, 1H), 4.72 (d, J = 15.0 Hz, 1H), 4.21 (d, J = 15.0 Hz, 1H), 3.80 (s, 3H), 3.62 - 3.74 (m, 3H).
¹³C NMR (CDCl₃, 100 MHz), δ159.4, 158.1, 131.4, 129.4, 128.3, 120.5, 114.3, 77.3, 59.2, 58.8, 55.3, 46.2. ¹ H NMR (CDCl ₃ , 400 MHz), δ7.26 (md, J = 8.7 Hz, 2H), 6.88 (md, J = 8.7 Hz, 2H), 6.05 (ddd, J = 6.9, 10.5, 17.2 Hz, 1H), 5.50 (md, J = 17.2 Hz, 1H), 5.40 (md, J = 10.6 Hz, 1H), 4.94 (mdd, J = 7.1, 8.0 Hz, 1H), 4.72 (d, J = 15.0 Hz, 1H), 4.21 (d, J = 15.0 Hz, 1H), 3.80 (s, 3H), 3.62-3.74 (m, 3H).
¹³ C NMR (CDCl ₃ , 100 MHz), δ159.4, 158.1, 131.4, 129.4, 128.3, 120.5, 114.3, 77.3, 59.2, 58.8, 55.3, 46.2.

（参考例１３）

(Reference Example 13)

(4S,5R)-4-ヒドロキシメチル-3-(4-メトキシベンジル)-5-ビニルオキサゾリジン-2-オン (1.900 g, 7.216 mmol)の塩化メチレン(35 ml)溶液に-78℃で2,6-ルチジン(1.01 ml, 8.66 mmol)、トリフルオロメタンスルホン酸無水物(1.46 ml, 8.66 mmol)を順次加え、10分間撹拌した。反応混合物を飽和炭酸水素ナトリウム水溶液中に注ぎ、中和した後クロロホルムで抽出した。有機層を飽和食塩水で洗浄し、無水硫酸マグネシウムで乾燥後、減圧濃縮を行った。残渣をシリカゲルクロマトグラフィー（ヘキサンに20〜33％の酢酸エチルを溶かしたもの）により精製し、トリフルオロメタンスルホン酸(4R,5R)-3-(4-メトキシベンジル)-2-オキソ-5-ビニルオキサゾリジン-4-イルメチルエステル(2.935 g, quant.)を得た。トリフルオロメタンスルホン酸(4R,5R)-3-(4-メトキシベンジル)-2-オキソ-5-ビニルオキサゾリジン-4-イルメチルエステルの¹H NMR、¹³C NMRデータを以下に示す。 (4S, 5R) -4-Hydroxymethyl-3- (4-methoxybenzyl) -5-vinyloxazolidine-2-one (1.900 g, 7.216 mmol) in methylene chloride (35 ml) at −78 ° C. 6-Lutidine (1.01 ml, 8.66 mmol) and trifluoromethanesulfonic anhydride (1.46 ml, 8.66 mmol) were sequentially added and stirred for 10 minutes. The reaction mixture was poured into a saturated aqueous sodium hydrogen carbonate solution, neutralized, and extracted with chloroform. The organic layer was washed with saturated brine, dried over anhydrous magnesium sulfate, and concentrated under reduced pressure. The residue was purified by silica gel chromatography (20-33% ethyl acetate in hexane) and trifluoromethanesulfonic acid (4R, 5R) -3- (4-methoxybenzyl) -2-oxo-5-vinyl Oxazolidin-4-ylmethyl ester (2.935 g, quant.) Was obtained. ¹ H NMR and ¹³ C NMR data of trifluoromethanesulfonic acid (4R, 5R) -3- (4-methoxybenzyl) -2-oxo-5-vinyloxazolidin-4-ylmethyl ester are shown below.

¹H NMR (CDCl₃, 400 MHz), δ7.23 (md, J = 8.7 Hz, 2H), 6.90 (md, J = 8.7 Hz, 2H), 5.85 (ddd, J = 6.4, 10.8, 17.1 Hz, 1H), 5.57 (ddd, J = 1.2, 1.2, 17.1 Hz, 1H), 5.50 (ddd, J = 1.2, 1.2, 10.8 Hz, 1H), 5.00 (dddd, J = 1.1, 1.2, 6.4, 8.5 Hz, 1H), 4.85 (d, J = 15.1 Hz, 1H), 4.48 (dd, J = 4.8, 10.8 Hz, 1H), 4.39 (dd, J = 4.8, 10.8 Hz, 1H), 4.10 (d, J = 15.1 Hz, 1H), 3.89 (td, J = 4.6, 8.2 Hz, 1H), 3.81 (s, 3H).
¹³C NMR (CDCl₃, 100 MHz), δ159.7, 156.9, 129.6, 129.0, 126.9, 122.0, 118.4 (q, J_C-F= 320.1 Hz), 71.7, 55.8, 55.3, 46.5. ¹ H NMR (CDCl ₃ , 400 MHz), δ7.23 (md, J = 8.7 Hz, 2H), 6.90 (md, J = 8.7 Hz, 2H), 5.85 (ddd, J = 6.4, 10.8, 17.1 Hz, 1H), 5.57 (ddd, J = 1.2, 1.2, 17.1 Hz, 1H), 5.50 (ddd, J = 1.2, 1.2, 10.8 Hz, 1H), 5.00 (dddd, J = 1.1, 1.2, 6.4, 8.5 Hz, 1H), 4.85 (d, J = 15.1 Hz, 1H), 4.48 (dd, J = 4.8, 10.8 Hz, 1H), 4.39 (dd, J = 4.8, 10.8 Hz, 1H), 4.10 (d, J = 15.1 Hz, 1H), 3.89 (td, J = 4.6, 8.2 Hz, 1H), 3.81 (s, 3H).
¹³ C NMR (CDCl ₃ , 100 MHz), δ159.7, 156.9, 129.6, 129.0, 126.9, 122.0, 118.4 (q, J _CF = 320.1 Hz), 71.7, 55.8, 55.3, 46.5.

（参考例１４）

(Reference Example 14)

メチルホスホン酸ジメチル(2.35 ml, 21.65 mmol)のTHF(35 ml)溶液に-78℃で1,6N n-ブチルリチウム(13.5 ml, 21.65 mmol)を滴下し、同温で30分間撹拌した。この混合物を-78℃に冷却したトリフルオロメタンスルホン酸(4R,5R)-3-(4-メトキシベンジル)-2-オキソ-5-ビニルオキサゾリジン-4-イルメチルエステル(2.935 g, 7.216 mmol)のTHF(35 ml)溶液中にTLCにて原料がほぼ消失するまで滴下した。反応混合物に飽和塩化アンモニウム水溶液を加え中和した後、酢酸エチルで抽出した。有機層を飽和食塩水で洗浄し、無水硫酸マグネシウムで乾燥後、減圧濃縮を行った。残渣をシリカゲルクロマトグラフィー（クロロホルムに0〜1.3％のメタノールを溶かしたもの）により精製し、{2-[(4S,5R)-3-(4-メトキシベンジル)-2-オキソ-5-ビニルオキサゾリジン-4-イル]-エチル}-ホスホン酸ジメチルエステル(1.826 g, 69% for 2 steps)を得た。{2-[(4S,5R)-3-(4-メトキシベンジル)-2-オキソ-5-ビニルオキサゾリジン-4-イル]-エチル}-ホスホン酸ジメチルエステルの¹H NMR、¹³C NMRデータを以下に示す。 To a solution of dimethyl methylphosphonate (2.35 ml, 21.65 mmol) in THF (35 ml) was added dropwise 1,6N n-butyllithium (13.5 ml, 21.65 mmol) at −78 ° C., and the mixture was stirred at the same temperature for 30 minutes. This mixture was cooled to −78 ° C. with trifluoromethanesulfonic acid (4R, 5R) -3- (4-methoxybenzyl) -2-oxo-5-vinyloxazolidin-4-ylmethyl ester (2.935 g, 7.216 mmol). The solution was added dropwise to a THF (35 ml) solution until almost disappearance of the raw material by TLC. The reaction mixture was neutralized with a saturated aqueous ammonium chloride solution, and extracted with ethyl acetate. The organic layer was washed with saturated brine, dried over anhydrous magnesium sulfate, and concentrated under reduced pressure. The residue was purified by silica gel chromatography (0 to 1.3% methanol in chloroform), and {2-[(4S, 5R) -3- (4-methoxybenzyl) -2-oxo-5-vinyloxazolidine -4-yl] -ethyl} -phosphonic acid dimethyl ester (1.826 g, 69% for 2 steps) was obtained. ¹ H NMR and ¹³ C NMR data of {2-[(4S, 5R) -3- (4-methoxybenzyl) -2-oxo-5-vinyloxazolidin-4-yl] -ethyl} -phosphonic acid dimethyl ester It is shown below.

¹H NMR (CDCl₃, 400 MHz), δ7.23 (md, J = 8.7 Hz, 2H), 6.88 (md, J = 8.7 Hz, 2H), 5.90 (ddd, J = 6.8, 10.5, 17.2 Hz, 1H), 5.51 (ddd, J = 1.1, 1.2, 17.2 Hz, 1H), 5.44 (mdd, J = 1.1, 10.5, Hz, 1H), 4.88 (mdd, J = 7.1, 7.8 Hz, 1H), 4.80 (d, J = 15.1 Hz, 1H), 3.97 (d, J = 15.1 Hz, 1H), 3.80 (s, 3H), 3.71 (d, J = 11.0 Hz, 3H), 3.70 (d, J = 10.7 Hz, 3H), 3.66 (dt, J = 3.2, 7.8 Hz, 1H), 1.75 - 1.95 (m, 2H), 1.59 - 1.68 (m, 2H).
¹³C NMR (CDCl₃, 100 MHz), δ159.3, 157.5, 130.2, 129.4, 127.5, 121.0, 114.2, 77.7, 56.6 (d, J_C-P= 17.7 Hz), 52.45 (d, J_C-P= 6.8 Hz), 52.40 (d, J_C-P= 6.8 Hz), 45.6, 20.6, 19.9 (d, J_C-P= 140 Hz). ¹ H NMR (CDCl ₃ , 400 MHz), δ7.23 (md, J = 8.7 Hz, 2H), 6.88 (md, J = 8.7 Hz, 2H), 5.90 (ddd, J = 6.8, 10.5, 17.2 Hz, 1H), 5.51 (ddd, J = 1.1, 1.2, 17.2 Hz, 1H), 5.44 (mdd, J = 1.1, 10.5, Hz, 1H), 4.88 (mdd, J = 7.1, 7.8 Hz, 1H), 4.80 ( d, J = 15.1 Hz, 1H), 3.97 (d, J = 15.1 Hz, 1H), 3.80 (s, 3H), 3.71 (d, J = 11.0 Hz, 3H), 3.70 (d, J = 10.7 Hz, 3H), 3.66 (dt, J = 3.2, 7.8 Hz, 1H), 1.75-1.95 (m, 2H), 1.59-1.68 (m, 2H).
¹³ C NMR (CDCl ₃ , 100 MHz), δ159.3, 157.5, 130.2, 129.4, 127.5, 121.0, 114.2, 77.7, 56.6 (d, J _CP = 17.7 Hz), 52.45 (d, J _CP = 6.8 Hz) , 52.40 (d, J _CP = 6.8 Hz), 45.6, 20.6, 19.9 (d, J _CP = 140 Hz).

（参考例１５）

(Reference Example 15)

{2-[(4S,5R)-3-(4-メトキシベンジル)-2-オキソ-5-ビニルオキサゾリジン-4-イル]-エチル}-ホスホン酸ジメチルエステル(2.733 g, 7.399 mmol)のアセトニトリル(22 ml)、水(7.4 ml)溶液中に0℃で硝酸二アンモニウムセリウム(12.170 g, 22.198 mmol)を加え同温で10分間撹拌した。反応混合物に飽和食塩水を加えた後、酢酸エチルで抽出した。有機層を飽和食塩水で洗浄し、無水硫酸マグネシウムで乾燥後、減圧濃縮を行った。残渣をシリカゲルクロマトグラフィー（クロロホルムに0〜4.8％のメタノールを溶かしたもの）により2回精製し、[2-((4S,5R)-2-オキソ-5-ビニルオキサゾリジン-4-イル)-エチル]-ホスホン酸ジメチルエステル(1.482 g, 80% yield)を得た。[2-((4S,5R)-2-オキソ-5-ビニルオキサゾリジン-4-イル)-エチル]-ホスホン酸ジメチルエステルの¹H NMR、¹³C NMRデータを以下に示す。 {2-[(4S, 5R) -3- (4-Methoxybenzyl) -2-oxo-5-vinyloxazolidin-4-yl] -ethyl} -phosphonic acid dimethyl ester (2.733 g, 7.399 mmol) in acetonitrile ( 22 ml) and water (7.4 ml) were added diammonium cerium nitrate (12.170 g, 22.198 mmol) at 0 ° C. and stirred at the same temperature for 10 minutes. Saturated brine was added to the reaction mixture, and the mixture was extracted with ethyl acetate. The organic layer was washed with saturated brine, dried over anhydrous magnesium sulfate, and concentrated under reduced pressure. The residue was purified twice by silica gel chromatography (0 to 4.8% methanol dissolved in chloroform) and [2-((4S, 5R) -2-oxo-5-vinyloxazolidine-4-yl) -ethyl ] -Phosphonic acid dimethyl ester (1.482 g, 80% yield) was obtained. ¹ H NMR and ¹³ C NMR data of [2-((4S, 5R) -2-oxo-5-vinyloxazolidin-4-yl) -ethyl] -phosphonic acid dimethyl ester are shown below.

¹H NMR (CDCl₃, 400 MHz), δ7.41 (brm, 1H), 5.90 (ddd, J = 6.9, 10.5, 17.2 Hz, 1H), 5.48 (mdd, J = 1,2, 17.2 Hz, 1H), 5.41 (mdd, 0.9, 10.5 Hz, 1H), 5.07 (dd, J = 7.4, 7.8 Hz, 1H), 3.94 (m, 1H), 3.75 (d, J = 10.8 Hz, 3H), 3.74 (d, J = 10.7 Hz, 3H), 1.65 - 2.13 (m, 4H).
¹³C NMR (CDCl₃, 100 MHz), δ158.9, 130.4, 120.6, 80.2, 55.8 (d, Jc-p = 13.4 Hz), 52.7 (d, Jc-p = 6.2 Hz), 52.7 (d, Jc-p = 6.3 Hz), 24.4 (d, Jc-p = 4.8 Hz), 21.2 (d, Jc-p = 142.8 Hz). ¹ H NMR (CDCl ₃ , 400 MHz), δ7.41 (brm, 1H), 5.90 (ddd, J = 6.9, 10.5, 17.2 Hz, 1H), 5.48 (mdd, J = 1,2, 17.2 Hz, 1H ), 5.41 (mdd, 0.9, 10.5 Hz, 1H), 5.07 (dd, J = 7.4, 7.8 Hz, 1H), 3.94 (m, 1H), 3.75 (d, J = 10.8 Hz, 3H), 3.74 (d , J = 10.7 Hz, 3H), 1.65-2.13 (m, 4H).
¹³ C NMR (CDCl ₃ , 100 MHz), δ158.9, 130.4, 120.6, 80.2, 55.8 (d, Jc-p = 13.4 Hz), 52.7 (d, Jc-p = 6.2 Hz), 52.7 (d, Jc -p = 6.3 Hz), 24.4 (d, Jc-p = 4.8 Hz), 21.2 (d, Jc-p = 142.8 Hz).

（参考例１６）

(Reference Example 16)

[2-((4S,5R)-2-オキソ-5-ビニルオキサゾリジン-4-イル)-エチル]-ホスホン酸ジメチルエステル (320 mg, 1.28 mmol)の塩化メチレン(13 ml)溶液に0℃で4-ジメチルアミノピリジン(78 mg, 0.64 mmol)トリエチルアミン(0.36 ml, 2.57 mmol)、Boc₂O(420 mg, 1.93 mmol)を順次加え、２０分間撹拌した。反応混合物に飽和塩化アンモニウム水溶液を加え中和した後、クロロホルムで抽出した。有機層を飽和食塩水で洗浄し、無水硫酸マグネシウムで乾燥後、減圧濃縮を行った。残渣をシリカゲルクロマトグラフィー（クロロホルムに0〜1％のメタノールを溶かしたもの）により精製し、(4S,5R)-4-[2-(ジメトキシホスホリル)-エチル]-2-オキソ-5-ビニルオキサゾリジン-3-カルボン酸tert-ブチルエステル(420 mg, 94% yield)を得た。(4S,5R)-4-[2-(ジメトキシホスホリル)-エチル]-2-オキソ-5-ビニルオキサゾリジン-3-カルボン酸tert-ブチルエステルの¹H NMR、¹³C NMRデータを以下に示す。 [2-((4S, 5R) -2-oxo-5-vinyloxazolidin-4-yl) -ethyl] -phosphonic acid dimethyl ester (320 mg, 1.28 mmol) in methylene chloride (13 ml) at 0 ° C. 4-Dimethylaminopyridine (78 mg, 0.64 mmol) triethylamine (0.36 ml, 2.57 mmol) and Boc ₂ O (420 mg, 1.93 mmol) were sequentially added and stirred for 20 minutes. The reaction mixture was neutralized with a saturated aqueous ammonium chloride solution and extracted with chloroform. The organic layer was washed with saturated brine, dried over anhydrous magnesium sulfate, and concentrated under reduced pressure. The residue was purified by silica gel chromatography (0 to 1% methanol in chloroform) and (4S, 5R) -4- [2- (dimethoxyphosphoryl) -ethyl] -2-oxo-5-vinyloxazolidine -3-Carboxylic acid tert-butyl ester (420 mg, 94% yield) was obtained. ¹ H NMR and ¹³ C NMR data of (4S, 5R) -4- [2- (dimethoxyphosphoryl) -ethyl] -2-oxo-5-vinyloxazolidine-3-carboxylic acid tert-butyl ester are shown below.

¹H NMR (CDCl₃, 400 MHz), δ5.90 (ddd, J = 6.6, 10.8, 17.2 Hz, 1H), 5.58 (md, J = 17.2 Hz, 1H), 5.51 (md, J = 10.8, Hz, 1H), 5.00 (m, 1H), 4.35 (dt, J = 4.8, 6.6 Hz, 1H), 3.741 (d, J = 10.8 Hz, 3H), 3.737 (d, J = 10.8 Hz, 3H), 2.02 - 2.14 (m, 1H), 1.84 - 1.97 (m, 1H), 1.70 - 1.82 (m, 2H), 1.55 (s, 9H).
¹³C NMR (CDCl₃, 100 MHz), δ151.3, 149.2, 129.0, 121.9, 84.3, 77.8, 58.1 (d, Jc-p = 20.1 Hz), 52.5 (d, Jc-p = 6.7 Hz), 52.46 (d, Jc-p = 6.7 Hz), 27.9, 22.5 (d, Jc-p = 3.8 Hz), 20.6 (d, Jc-p = 142.8 Hz). ¹ H NMR (CDCl ₃ , 400 MHz), δ5.90 (ddd, J = 6.6, 10.8, 17.2 Hz, 1H), 5.58 (md, J = 17.2 Hz, 1H), 5.51 (md, J = 10.8, Hz , 1H), 5.00 (m, 1H), 4.35 (dt, J = 4.8, 6.6 Hz, 1H), 3.741 (d, J = 10.8 Hz, 3H), 3.737 (d, J = 10.8 Hz, 3H), 2.02 -2.14 (m, 1H), 1.84-1.97 (m, 1H), 1.70-1.82 (m, 2H), 1.55 (s, 9H).
¹³ C NMR (CDCl ₃ , 100 MHz), δ151.3, 149.2, 129.0, 121.9, 84.3, 77.8, 58.1 (d, Jc-p = 20.1 Hz), 52.5 (d, Jc-p = 6.7 Hz), 52.46 (d, Jc-p = 6.7 Hz), 27.9, 22.5 (d, Jc-p = 3.8 Hz), 20.6 (d, Jc-p = 142.8 Hz).

（参考例１７）

(Reference Example 17)

(4S,5R)-4-[2-(ジメトキシホスホリル)-エチル]-2-オキソ-5-ビニルオキサゾリジン-3-カルボン酸tert-ブチルエステル(100 mg, 0.286 mmol)のメタノール(2.9 ml)溶液に室温で炭酸カリウム(0.198 g, 1.431 mmol)を加え、１２分間撹拌した。反応混合物に飽和塩化アンモニウム水溶液を加え中和した後、酢酸エチルで抽出した。有機層を飽和食塩水で洗浄し、無水硫酸マグネシウムで乾燥後、減圧濃縮を行った。残渣をシリカゲルクロマトグラフィー（クロロホルムに0〜2％のメタノールを溶かしたもの）により精製し、(3S,4R)-(3-tert-ブトキシカルボニルアミノ-4-ヒドロキシヘキサ-5-エニル)ホスホン酸ジメチルエステル(59 mg, 64% yield)を得た。(3S,4R)-(3-tert-ブトキシカルボニルアミノ-4-ヒドロキシヘキサ-5-エニル)ホスホン酸ジメチルエステルの¹H NMR、¹³C NMRデータを以下に示す。 (4S, 5R) -4- [2- (Dimethoxyphosphoryl) -ethyl] -2-oxo-5-vinyloxazolidine-3-carboxylic acid tert-butyl ester (100 mg, 0.286 mmol) in methanol (2.9 ml) Was added potassium carbonate (0.198 g, 1.431 mmol) at room temperature and stirred for 12 minutes. The reaction mixture was neutralized with a saturated aqueous ammonium chloride solution, and extracted with ethyl acetate. The organic layer was washed with saturated brine, dried over anhydrous magnesium sulfate, and concentrated under reduced pressure. The residue was purified by silica gel chromatography (0-2% methanol in chloroform) and dimethyl (3S, 4R)-(3-tert-butoxycarbonylamino-4-hydroxyhex-5-enyl) phosphonate An ester (59 mg, 64% yield) was obtained. ¹ H NMR and ¹³ C NMR data of (3S, 4R)-(3-tert-butoxycarbonylamino-4-hydroxyhex-5-enyl) phosphonic acid dimethyl ester are shown below.

¹H NMR (CDCl₃, 400 MHz), δ5.86 (ddd, J = 5.5, 10.5, 17.2 Hz, 1H), 5.36 (ddd, J = 1.4, 1.6, 17.2 Hz, 1H), 5.23 (ddd, J = 1.4, 1.4, 10.5 Hz, 1H), 5.13 (brd, J = 9.2 Hz, 1H), 4.23 (m, 1H), 3.73 (d, J = 10.8 Hz, 6H), 3.67 (m, 1H), 2.22 (brm, 1H), 1.60 - 1.92 (m, 4H), 1.45 (s, 9H).
¹³C NMR (CDCl₃, 100 MHz), δ156.5, 136.9, 116.8, 79.8, 75.1, 55.4 (d, Jc-p = 16.3 Hz), 52.4 (d, Jc-p = 6.7 Hz), 28.3, 22.4, 21.5 (d, Jc-p = 142.3 Hz). ¹ H NMR (CDCl ₃ , 400 MHz), δ5.86 (ddd, J = 5.5, 10.5, 17.2 Hz, 1H), 5.36 (ddd, J = 1.4, 1.6, 17.2 Hz, 1H), 5.23 (ddd, J = 1.4, 1.4, 10.5 Hz, 1H), 5.13 (brd, J = 9.2 Hz, 1H), 4.23 (m, 1H), 3.73 (d, J = 10.8 Hz, 6H), 3.67 (m, 1H), 2.22 (brm, 1H), 1.60-1.92 (m, 4H), 1.45 (s, 9H).
¹³ C NMR (CDCl ₃ , 100 MHz), δ156.5, 136.9, 116.8, 79.8, 75.1, 55.4 (d, Jc-p = 16.3 Hz), 52.4 (d, Jc-p = 6.7 Hz), 28.3, 22.4 , 21.5 (d, Jc-p = 142.3 Hz).

（実施例１）

Example 1

[2-((4R,5R)-3-t-ブトキシカルボニル-2-オキソ-5-ビニルオキサゾリジン-4-イル)エチル] ホスホン酸ジメチルエステル (0.05 g, 0.143 mmol)の塩化メチレン(2.9 ml)溶液に11-[(7-ニトロベンゾ[1,2,5]オキサジアゾール-4-イル)アミノ]-1-ウンデセン (0.143 g, 0.429 mmol)、1,3-(ビス(メシチル)-2-イミダゾリジニリデン)ジクロロ(フェニルメチレン)(トリシクロヘキシルホスフィン)ルテニウム（Grubbs 2nd catalyst：0.004 g, 0.004 mmol)を順次加え、２時間加熱還流した。反応混合物を減圧濃縮し、残渣をシリカゲルクロマトグラフィー(クロロホルムに0〜４%のメタノールを溶かしたもの)により粗精製し、(2-((4R,5R)-3-t-ブトキシカルボニル-5-[11-(7-ニトロベンゾ[1,2,5]オキサジアゾール-4-イルアミノ)-ウンデカ-1-エニル]-2-オキソオキサゾリジン-4-イル）エチル）ホスホン酸ジメチルエステルを[2-(3-t-ブトキシカルボニル-2-オキソ-5-ビニルオキサゾリジン-4-イル)エチル] ホスホン酸ジメチルエステルとの混合物として得た(0.089 g)。(2-((4R,5R)-3-t-ブトキシカルボニル-5-[11-(7-ニトロベンゾ[1,2,5]オキサジアゾール-4-イルアミノ)-ウンデカ-1-エニル]-2-オキソオキサゾリジン-4-イル）エチル）ホスホン酸ジメチルエステルの¹H NMR、¹³C NMRデータを以下に示す。 [2-((4R, 5R) -3-t-Butoxycarbonyl-2-oxo-5-vinyloxazolidine-4-yl) ethyl] phosphonic acid dimethyl ester (0.05 g, 0.143 mmol) in methylene chloride (2.9 ml) 11-[(7-nitrobenzo [1,2,5] oxadiazol-4-yl) amino] -1-undecene (0.143 g, 0.429 mmol), 1,3- (bis (mesityl) -2- Imidazolidinylidene) dichloro (phenylmethylene) (tricyclohexylphosphine) ruthenium (Grubbs 2nd catalyst: 0.004 g, 0.004 mmol) was sequentially added and heated to reflux for 2 hours. The reaction mixture was concentrated under reduced pressure, and the residue was roughly purified by silica gel chromatography (0-4% methanol dissolved in chloroform) to give (2-((4R, 5R) -3-t-butoxycarbonyl-5- [11- (7-Nitrobenzo [1,2,5] oxadiazol-4-ylamino) -undec-1-enyl] -2-oxooxazolidin-4-yl) ethyl) phosphonic acid dimethyl ester [2- ( 3-t-Butoxycarbonyl-2-oxo-5-vinyloxazolidine-4-yl) ethyl] obtained as a mixture with phosphonic acid dimethyl ester (0.089 g). (2-((4R, 5R) -3-t-butoxycarbonyl-5- [11- (7-nitrobenzo [1,2,5] oxadiazol-4-ylamino) -undec-1-enyl] -2 ¹ H NMR and ¹³ C NMR data of -oxooxazolidin-4-yl) ethyl) phosphonic acid dimethyl ester are shown below.

¹H NMR (CDCl₃, 400 MHz) δ8.50 (d, J = 8.8 Hz, 1H), 6.59 (br m, 1H), 6.18 (d, J = 8.8 Hz, 1H), 5.87 (td, J = 6.8, 14.9 Hz, 1H), 5.49 (dd, J = 7.1, 15.6 Hz, 1H), 4.51 (dd, J = 3.7, 7.1 Hz, 1H), 4.01 (m, 1H), 3.76 (d, J = 10.7 Hz, 6H), 3.50 (m, 2H), 1.70 - 2.14 (m, 6H), 1.55 (s, 9H), 1.29 - 1.47 (m, 14 H)
¹³C NMR (CDCl₃, 100 MHz) δ151.5, 149.3, 144.3, 144.0, 137.5, 136.5, 125.5, 98.4, 84.3, 78.1, 60.1 (d, Jc-p = 18.2 Hz), 52.6 (m), 44.0, 31.9, 29.2, 29.1, 29.0, 28.8, 28.4, 28.3, 27.9, 26.8, 27.9, 25.5 (d, Jc-p = 4.1 Hz), 19.6 (d, Jc-p = 144.7 Hz) ¹ H NMR (CDCl ₃ , 400 MHz) δ8.50 (d, J = 8.8 Hz, 1H), 6.59 (br m, 1H), 6.18 (d, J = 8.8 Hz, 1H), 5.87 (td, J = 6.8, 14.9 Hz, 1H), 5.49 (dd, J = 7.1, 15.6 Hz, 1H), 4.51 (dd, J = 3.7, 7.1 Hz, 1H), 4.01 (m, 1H), 3.76 (d, J = 10.7 Hz, 6H), 3.50 (m, 2H), 1.70-2.14 (m, 6H), 1.55 (s, 9H), 1.29-1.47 (m, 14 H)
¹³ C NMR (CDCl ₃ , 100 MHz) δ151.5, 149.3, 144.3, 144.0, 137.5, 136.5, 125.5, 98.4, 84.3, 78.1, 60.1 (d, Jc-p = 18.2 Hz), 52.6 (m), 44.0 , 31.9, 29.2, 29.1, 29.0, 28.8, 28.4, 28.3, 27.9, 26.8, 27.9, 25.5 (d, Jc-p = 4.1 Hz), 19.6 (d, Jc-p = 144.7 Hz)

（実施例２）

(Example 2)

(2-((4R,5R)-3-t-ブトキシカルボニル-5-[11-(7-ニトロベンゾ[1,2,5]オキサジアゾール-4-イルアミノ)-ウンデカ-1-エニル]-2-オキソオキサゾリジン-4-イル）エチル）ホスホン酸ジメチルエステル (0.374 g, 0.572 mmol)のMeOH(5.7 ml)溶液に0 ℃で炭酸カリウム(0.395 g, 2.861 mmol)を加え、室温で1時間攪拌した。反応混合物に飽和塩化アンモニウム水溶液を加え、クロロホルムで抽出した。有機層を飽和食塩水溶液で洗浄し、無水硫酸マグネシウムで乾燥後、減圧濃縮を行った。残渣をシリカゲルクロマトグラフィー(クロロホルムに9〜17%のアセトンを溶かしたもの）により精製し、[(3R,4R,5E)-3-t-ブトキシカルボニルアミノ-4-ヒドロキシ-15-(7-ニトロベンゾ[1,2,5]オキサジアゾール-4-イルアミノ）ペンタデカ-5-エニル]ホスホン酸ジメチルエステルを得た(0.133 g, 37% yield)。[(3R,4R,5E)-3-t-ブトキシカルボニルアミノ-4-ヒドロキシ-15-(7-ニトロベンゾ[1,2,5]オキサジアゾール-4-イルアミノ）ペンタデカ-5-エニル]ホスホン酸ジメチルエステルの¹H NMR、¹³C NMRデータを以下に示す。 (2-((4R, 5R) -3-t-butoxycarbonyl-5- [11- (7-nitrobenzo [1,2,5] oxadiazol-4-ylamino) -undec-1-enyl] -2 -Oxooxazolidin-4-yl) ethyl) phosphonic acid dimethyl ester (0.374 g, 0.572 mmol) in MeOH (5.7 ml) was added potassium carbonate (0.395 g, 2.861 mmol) at 0 ° C. and stirred at room temperature for 1 hour. . A saturated aqueous ammonium chloride solution was added to the reaction mixture, and the mixture was extracted with chloroform. The organic layer was washed with a saturated aqueous sodium chloride solution, dried over anhydrous magnesium sulfate, and concentrated under reduced pressure. The residue was purified by silica gel chromatography (9-17% acetone in chloroform) and [(3R, 4R, 5E) -3-t-butoxycarbonylamino-4-hydroxy-15- (7-nitrobenzo [1,2,5] oxadiazol-4-ylamino) pentadeca-5-enyl] phosphonic acid dimethyl ester was obtained (0.133 g, 37% yield). [(3R, 4R, 5E) -3-t-Butoxycarbonylamino-4-hydroxy-15- (7-nitrobenzo [1,2,5] oxadiazol-4-ylamino) pentadeca-5-enyl] phosphonic acid ¹ H NMR and ¹³ C NMR data of dimethyl ester are shown below.

¹H NMR (CDCl₃, 400 MHz) δ8.48 (d, J = 8.8 Hz, 1H), 7.08 (br m, 1H), 6.18 (d, J = 8.8 Hz, 1H), 5.71 (td, J = 7.1, 15.5 Hz, 1H), 5.47 (dd, J = 6.6, 15.6 Hz, 1H), 5.01 (br d, J = 7.8 Hz, 1H), 4.09 (m, 1H), 3.73 (d, J = 10.7 Hz, 6H), 3.50 - 3.59 (m, 3H), 2.01 (m, 2H), 1.78 - 1.91 (m, 4H), 1.43 (s, 9H), 1.28 - 1.38 (m, 14 H)
¹³C NMR (CDCl₃, 100 MHz)δ156.3, 144.2, 143.9, 136.6, 133.4, 129.4, 98.4, 79.2, 73.7, 55.3 (d, Jc-p = 17.4 Hz), 52.4 (d, Jc-p = 6.6 Hz), 52.3 (d, Jc-p = 6.6 Hz), 45.7, 44.0, 32.1, 29.2, 29.1, 29.0, 28.9, 28.8, 28.3, 26.8, 24.8 (m), 21.2 (d, Jc-p = 141.4 Hz) ¹ H NMR (CDCl ₃ , 400 MHz) δ8.48 (d, J = 8.8 Hz, 1H), 7.08 (br m, 1H), 6.18 (d, J = 8.8 Hz, 1H), 5.71 (td, J = 7.1, 15.5 Hz, 1H), 5.47 (dd, J = 6.6, 15.6 Hz, 1H), 5.01 (br d, J = 7.8 Hz, 1H), 4.09 (m, 1H), 3.73 (d, J = 10.7 Hz , 6H), 3.50-3.59 (m, 3H), 2.01 (m, 2H), 1.78-1.91 (m, 4H), 1.43 (s, 9H), 1.28-1.38 (m, 14 H)
¹³ C NMR (CDCl ₃ , 100 MHz) δ156.3, 144.2, 143.9, 136.6, 133.4, 129.4, 98.4, 79.2, 73.7, 55.3 (d, Jc-p = 17.4 Hz), 52.4 (d, Jc-p = 6.6 Hz), 52.3 (d, Jc-p = 6.6 Hz), 45.7, 44.0, 32.1, 29.2, 29.1, 29.0, 28.9, 28.8, 28.3, 26.8, 24.8 (m), 21.2 (d, Jc-p = 141.4 Hz)

（実施例３）

(Example 3)

[(3R,4R,5E)-3-t-ブトキシカルボニルアミノ-4-ヒドロキシ-15-(7-ニトロベンゾ[1,2,5]オキサジアゾール-4-イルアミノ）ペンタデカ-5-エニル]ホスホン酸ジメチルエステル (0.077 g, 0.123 mmol)のDMF(1.23 ml)溶液に室温でイミダゾール(0.017 g, 0.246 mmol)、クロロトリエチルシラン(0.031 ml, 0.185 mmol)を加え、10分間攪拌した。反応混合物に飽和塩化アンモニウム水溶液を加え、酢酸エチルで抽出した。有機層を飽和食塩水溶液で洗浄し、無水硫酸マグネシウムで乾燥後、減圧濃縮を行った。残渣をシリカゲルクロマトグラフィー(トリエチルアミン３％を含むヘキサンに50〜67%の酢酸エチルを溶かしたもの)により精製し、[(3R,4R,5E)-3-t-ブトキシカルボニルアミノ-4-トリエチルシリルオキシ-15-(7-ニトロベンゾ[1,2,5]オキサジアゾール-4-イルアミノ）ペンタデカ-5-エニル]ホスホン酸ジメチルエステルを得た(0.092 g, quant.)。[(3R,4R,5E)-3-t-ブトキシカルボニルアミノ-4-トリエチルシリルオキシ-15-(7-ニトロベンゾ[1,2,5]オキサジアゾール-4-イルアミノ）ペンタデカ-5-エニル]ホスホン酸ジメチルエステルのIR、¹H NMR、¹³C NMRデータを以下に示す。 [(3R, 4R, 5E) -3-t-Butoxycarbonylamino-4-hydroxy-15- (7-nitrobenzo [1,2,5] oxadiazol-4-ylamino) pentadeca-5-enyl] phosphonic acid Imidazole (0.017 g, 0.246 mmol) and chlorotriethylsilane (0.031 ml, 0.185 mmol) were added to a DMF (1.23 ml) solution of dimethyl ester (0.077 g, 0.123 mmol) at room temperature, and the mixture was stirred for 10 minutes. A saturated aqueous ammonium chloride solution was added to the reaction mixture, and the mixture was extracted with ethyl acetate. The organic layer was washed with a saturated aqueous sodium chloride solution, dried over anhydrous magnesium sulfate, and concentrated under reduced pressure. The residue was purified by silica gel chromatography (50-67% ethyl acetate in hexane containing 3% triethylamine), and [(3R, 4R, 5E) -3-t-butoxycarbonylamino-4-triethylsilyl Oxy-15- (7-nitrobenzo [1,2,5] oxadiazol-4-ylamino) pentadeca-5-enyl] phosphonic acid dimethyl ester was obtained (0.092 g, quant.). [(3R, 4R, 5E) -3-t-butoxycarbonylamino-4-triethylsilyloxy-15- (7-nitrobenzo [1,2,5] oxadiazol-4-ylamino) pentadeca-5-enyl] IR, ¹ H NMR and ¹³ C NMR data of phosphonic acid dimethyl ester are shown below.

IR (KBr disk): 3221, 2926, 2855, 1620, 1588, 1302, 1273, 1036, 816 cm^-1
¹H NMR (CDCl₃, 400 MHz) δ8.48 (d, J = 8.8 Hz, 1H), 6.89 (br m, 1H), 6.18 (d, J = 8.8 Hz, 1H), 5.61 (td, J = 6.6, 15.1 Hz, 1H), 5.39 (dd, J = 7.1, 15.1 Hz, 1H), 4.65 (br d, J = 9.5 Hz, 1H), 4.07 (m, 1H), 3.73 (d, J = 10.7 Hz, 6H), 3.52 (m, 3H), 2.00 (m, 2H), 1.59 - 1.90 (m, 6H), 1.43 (s, 9H), 1.29 - 1.48 (m, 12 H), 0.93 (t, J = 7.8 z, 9H), 0.58 (m, 6H)
¹³C NMR (CDCl₃, 100 MHz)δ155.9, 144.22, 144.15, 143.9, 136.5, 133.0, 129.6, 123.4, 98.4, 79.0, 74.8, 56.0 (d, Jc-p = 18.2 Hz), 52.3 (d, Jc-p = 5.8 Hz), 44.0, 32.0, 29.3, 29.2, 29.1, 29.0, 28.8, 28.4, 28.3, 26.9, 25.0, 21.6 (d, Jc-p = 141.4 Hz), 6.73, 4.87 IR (KBr disk): 3221, 2926, 2855, 1620, 1588, 1302, 1273, 1036, 816 cm ^-1
¹ H NMR (CDCl ₃ , 400 MHz) δ8.48 (d, J = 8.8 Hz, 1H), 6.89 (br m, 1H), 6.18 (d, J = 8.8 Hz, 1H), 5.61 (td, J = 6.6, 15.1 Hz, 1H), 5.39 (dd, J = 7.1, 15.1 Hz, 1H), 4.65 (br d, J = 9.5 Hz, 1H), 4.07 (m, 1H), 3.73 (d, J = 10.7 Hz , 6H), 3.52 (m, 3H), 2.00 (m, 2H), 1.59-1.90 (m, 6H), 1.43 (s, 9H), 1.29-1.48 (m, 12 H), 0.93 (t, J = 7.8 z, 9H), 0.58 (m, 6H)
¹³ C NMR (CDCl ₃ , 100 MHz) δ155.9, 144.22, 144.15, 143.9, 136.5, 133.0, 129.6, 123.4, 98.4, 79.0, 74.8, 56.0 (d, Jc-p = 18.2 Hz), 52.3 (d, Jc-p = 5.8 Hz), 44.0, 32.0, 29.3, 29.2, 29.1, 29.0, 28.8, 28.4, 28.3, 26.9, 25.0, 21.6 (d, Jc-p = 141.4 Hz), 6.73, 4.87

（実施例４）

Example 4

[(3R,4R,5E)-3-t-ブトキシカルボニルアミノ-4-トリエチルシリルオキシ-15-(7-ニトロベンゾ[1,2,5]オキサジアゾール-4-イルアミノ）ペンタデカ-5-エニル]ホスホン酸ジメチルエステル (0.032 g, 0.043 mmol)の塩化メチレン(0.86 ml)溶液に0 ℃でトリフルオロ酢酸(0.129 ml)を加え、室温で２時間攪拌した。反応混合物に飽和炭酸水素ナトリウム水溶液を加え、クロロホルムで抽出した。有機層を飽和食塩水溶液で洗浄し、減圧濃縮を行った。残渣をシリカゲルクロマトグラフィー(クロロホルムに9〜26.6%のメタノールと4.3%の水を加えたもの)により粗精製し、[(3R,4R,5E)-3-アミノ-4-ヒドロキシ-15-(7-ニトロベンゾ[1,2,5]オキサジアゾール-4-イルアミノ）ペンタデカ-5-エニル]ホスホン酸ジメチルエステルを得た(0.010 g, 44% yield)。[(3R,4R,5E)-3-アミノ-4-ヒドロキシ-15-(7-ニトロベンゾ[1,2,5]オキサジアゾール-4-イルアミノ）ペンタデカ-5-エニル]ホスホン酸ジメチルエステルの¹H NMRデータを以下に示す。 [(3R, 4R, 5E) -3-t-butoxycarbonylamino-4-triethylsilyloxy-15- (7-nitrobenzo [1,2,5] oxadiazol-4-ylamino) pentadeca-5-enyl] Trifluoroacetic acid (0.129 ml) was added to a methylene chloride (0.86 ml) solution of phosphonic acid dimethyl ester (0.032 g, 0.043 mmol) at 0 ° C., and the mixture was stirred at room temperature for 2 hours. A saturated aqueous sodium hydrogen carbonate solution was added to the reaction mixture, and the mixture was extracted with chloroform. The organic layer was washed with a saturated saline solution and concentrated under reduced pressure. The residue was roughly purified by silica gel chromatography (9-26.6% methanol and 4.3% water added to chloroform) to give [(3R, 4R, 5E) -3-amino-4-hydroxy-15- (7 -Nitrobenzo [1,2,5] oxadiazol-4-ylamino) pentadeca-5-enyl] phosphonic acid dimethyl ester was obtained (0.010 g, 44% yield). Of [(3R, 4R, 5E) -3- amino-4-hydroxy-15- (7-nitrobenzo [1,2,5] oxadiazol-4-ylamino) pentadeca-5-enyl] phosphonic acid dimethyl ester ¹ 1 H NMR data is shown below.

¹H NMR (CD₃OD, 400 MHz) δ8.50 (d, J = 8.5 Hz, 1H), 6.33 (d, J = 8.8 Hz, 1H), 5.73 (m, J = 1H), 5.44 (dd, J = 7.6, 15.4 Hz, 1H), 3.80 (m, 1H), 3.74 (d, J = 10.7 Hz, 6H), 3.52 (m, 2H), 2.82 - 3.02 (m, 1H), 2.68 (br m, 1H), 2.05 (m, 2H), 1.79 (m, 2H), 1.32 - 2.05 (m, 18 H) ¹ H NMR (CD ₃ OD, 400 MHz) δ8.50 (d, J = 8.5 Hz, 1H), 6.33 (d, J = 8.8 Hz, 1H), 5.73 (m, J = 1H), 5.44 (dd, J = 7.6, 15.4 Hz, 1H), 3.80 (m, 1H), 3.74 (d, J = 10.7 Hz, 6H), 3.52 (m, 2H), 2.82-3.02 (m, 1H), 2.68 (br m, 1H), 2.05 (m, 2H), 1.79 (m, 2H), 1.32-2.05 (m, 18 H)

（実施例５）

(Example 5)

[(3R,4R,5E)-3-アミノ-4-ヒドロキシ-15-(7-ニトロベンゾ[1,2,5]オキサジアゾール-4-イルアミノ）ペンタデカ-5-エニル]ホスホン酸ジメチルエステル(0.03 g, 0.040 mmol)のアセトニトリル(1.0 ml)溶液に0 ℃でブロモトリメチルシラン(0.054 ml, 0.404 mmol)を加え、室温で30分間攪拌した。反応混合物を減圧濃縮し、残渣に室温でメタノール（1.5 ml）を加え、同温で1時間攪拌した。反応混合物を減圧濃縮し、残渣を逆相シリカゲルクロマトグラフィー(メタノールに33%の水を加えたもの)により粗精製し、[(3R,4R,5E)-3-アミノ-4-ヒドロキシ-15-(7-ニトロベンゾ[1,2,5]オキサジアゾール-4-イルアミノ）ペンタデカ-5-エニル]ホスホン酸を得た(0.023 g, quant.)。これを逆相HPLC(溶離液：メタノールに９％の水を加えたもの、流速：2 ml/min、保持時間：７分、カラム：Develosil C8-5 (250 x 10 mmI.D.)、検出波長：220nm)により精製した( 0.018 g, 78% yield)。[(3R,4R,5E)-3-アミノ-4-ヒドロキシ-15-(7-ニトロベンゾ[1,2,5]オキサジアゾール-4-イルアミノ）ペンタデカ-5-エニル]ホスホン酸のIR、¹H NMR、¹³C NMRデータを以下に示す。 [(3R, 4R, 5E) -3-Amino-4-hydroxy-15- (7-nitrobenzo [1,2,5] oxadiazol-4-ylamino) pentadeca-5-enyl] phosphonic acid dimethyl ester (0.03 g, 0.040 mmol) in acetonitrile (1.0 ml) was added bromotrimethylsilane (0.054 ml, 0.404 mmol) at 0 ° C. and stirred at room temperature for 30 minutes. The reaction mixture was concentrated under reduced pressure, methanol (1.5 ml) was added to the residue at room temperature, and the mixture was stirred at the same temperature for 1 hr. The reaction mixture was concentrated under reduced pressure, and the residue was roughly purified by reverse-phase silica gel chromatography (methanol added with 33% water) to give [(3R, 4R, 5E) -3-amino-4-hydroxy-15- (7-Nitrobenzo [1,2,5] oxadiazol-4-ylamino) pentadeca-5-enyl] phosphonic acid was obtained (0.023 g, quant.). This was reverse-phase HPLC (eluent: methanol plus 9% water, flow rate: 2 ml / min, retention time: 7 minutes, column: Develosil C8-5 (250 x 10 mm I.D.), detection (Wavelength: 220 nm) (0.018 g, 78% yield). IR of [(3R, 4R, 5E) -3-amino-4-hydroxy-15- (7-nitrobenzo [1,2,5] oxadiazol-4-ylamino) pentadeca-5-enyl] phosphonic acid, ¹ 1 H NMR and ¹³ C NMR data are shown below.

IR (KBr disk): 3455, 2930, 1655, 1588, 1402, 1298, 1256 1088 cm^-1
¹H NMR (CD₃OD, 400 MHz) δ8.53 (d, J = 9.0 Hz, 1H), 6.38 (d, J = 9.0 Hz, 1H), 5.83 (dtd, J = 0.98, 6.8, 15.4 Hz, 1H), 5.44 (tdd, J = 1.5, 7.3, 15.4 Hz, 1H), 3.99 (m, 1H), 3.74 (d, J = 10.7 Hz, 6H), 3.52 (m, 2H), 3.04 (m, 1H), 2.08, (m, 2H), 1.92 - 2.04 (m, 1H), 1.58 - 1.82 (m, 5H), 1.32 - 1.48 (m, 12 H)
¹³C NMR (CD₃OD, 100 MHz) δ145.6, 137.2, 135.8, 128.2, 123.5, 98.2, 71.3, 56.3(d, Jc-p = 14.1 Hz), 43.4, 31.9, 29.1, 29.0, 28.90, 28.88, 28.6, 27.8, 26.6, 24.7, 24.6, 26.6, 23.1 (d, Jc-p = 3.3 Hz), 23.0 (d, Jc-p = 142.3 Hz) IR (KBr disk): 3455, 2930, 1655, 1588, 1402, 1298, 1256 1088 cm ^-1
¹ H NMR (CD ₃ OD, 400 MHz) δ8.53 (d, J = 9.0 Hz, 1H), 6.38 (d, J = 9.0 Hz, 1H), 5.83 (dtd, J = 0.98, 6.8, 15.4 Hz, 1H), 5.44 (tdd, J = 1.5, 7.3, 15.4 Hz, 1H), 3.99 (m, 1H), 3.74 (d, J = 10.7 Hz, 6H), 3.52 (m, 2H), 3.04 (m, 1H ), 2.08, (m, 2H), 1.92-2.04 (m, 1H), 1.58-1.82 (m, 5H), 1.32-1.48 (m, 12 H)
¹³ C NMR (CD ₃ OD, 100 MHz) δ145.6, 137.2, 135.8, 128.2, 123.5, 98.2, 71.3, 56.3 (d, Jc-p = 14.1 Hz), 43.4, 31.9, 29.1, 29.0, 28.90, 28.88 , 28.6, 27.8, 26.6, 24.7, 24.6, 26.6, 23.1 (d, Jc-p = 3.3 Hz), 23.0 (d, Jc-p = 142.3 Hz)

（実施例６）

(Example 6)

(3S,4R)-(3-tert-ブトキシカルボニルアミノ-4-ヒドロキシヘキサ-5-エニル)ホスホン酸ジメチルエステル(258 mg, 0.797 mmol)の塩化メチレン(8.0 ml)溶液に室温で11-[(7-ニトロベンゾ[1,2,5]オキサジアゾール-4-イル)アミノ]-1-ウンデセン(1.061 g, 3.19 mmol)、1,3-(ビス(メシチル)-2-イミダゾリジニリデン)ジクロロ(フェニルメチレン)(トリシクロヘキシルホスフィン)ルテニウム(Grubbs 2nd catalyst) (24 mg, 0.024 mmol)を加え、４０℃で3時間還流した。反応混合物に水を加え、クロロホルムで抽出した。有機層を飽和食塩水で洗浄し、無水硫酸マグネシウムで乾燥後、減圧濃縮を行った。残渣をシリカゲルクロマトグラフィー（クロロホルムに0〜2％のメタノールを溶かしたもの）により精製し、[(3S,4R,5E)-3-tert-ブトキシアルボニルアミノ-4-ヒドロキシ-15-(7-ニトロベンゾ[1,2,5]オキサジアゾール-4-イルアミノ)-ベンタデカ-5-エニル]-ホスホン酸ジメチルエステル(428 mg, 85% yield)を得た。[(3S,4R,5E)-3-tert-ブトキシアルボニルアミノ-4-ヒドロキシ-15-(7-ニトロベンゾ[1,2,5]オキサジアゾール-4-イルアミノ)-ベンタデカ-5-エニル]-ホスホン酸ジメチルエステルの¹H NMR、¹³C NMRデータを以下に示す。 11-[((3S, 4R)-(3-tert-butoxycarbonylamino-4-hydroxyhex-5-enyl) phosphonic acid dimethyl ester (258 mg, 0.797 mmol) in methylene chloride (8.0 ml) at room temperature. 7-nitrobenzo [1,2,5] oxadiazol-4-yl) amino] -1-undecene (1.061 g, 3.19 mmol), 1,3- (bis (mesityl) -2-imidazolidinylidene) dichloro ( Phenylmethylene) (tricyclohexylphosphine) ruthenium (Grubbs 2nd catalyst) (24 mg, 0.024 mmol) was added, and the mixture was refluxed at 40 ° C. for 3 hours. Water was added to the reaction mixture, and the mixture was extracted with chloroform. The organic layer was washed with saturated brine, dried over anhydrous magnesium sulfate, and concentrated under reduced pressure. The residue was purified by silica gel chromatography (0 to 2% methanol in chloroform) and [(3S, 4R, 5E) -3-tert-butoxycarbonylamino-4-hydroxy-15- (7- Nitrobenzo [1,2,5] oxadiazol-4-ylamino) -bentadeca-5-enyl] -phosphonic acid dimethyl ester (428 mg, 85% yield) was obtained. [(3S, 4R, 5E) -3-tert-butoxycarbonylamino-4-hydroxy-15- (7-nitrobenzo [1,2,5] oxadiazol-4-ylamino) -bentadeca-5-enyl] ^-1 H NMR and ¹³ C NMR data of phosphonic acid dimethyl ester are shown below.

¹H NMR (CDCl₃, 400 MHz), δ8.49 (d, J = 8.7 Hz, 2H), 6.76 (brm, 1H), 6.18 (d, J = 8.7 Hz, 2H), 5.72 (td, J = 6.6, 15.6 Hz, 1H), 5.45 (dd, J = 15.3 Hz, 1H), 4.92 (brd, J = 8.5 Hz, 1H), 4.15 (m, 1H), 3.74 (d, J = 11.0 Hz, 6H), 3.63 (m, 1H), 3.50 (td, J = 6.4, 6.4 Hz, 2H), 2.04 (dt, J = 6.6, 6.9 Hz, 2H), 1.60 - 1.92 (m, 8H), 1.44 (s, 9H), 1.29 - 1.48 (m, 12H).
¹³C NMR (CDCl₃, 100 MHz), δ156.4, 144.3, 144.09, 143.95, 136.6, 134.1, 228.5, 123.6, 98.5, 79.7, 75.1, 55.7 (d, Jc-p = 13.4 Hz), 52.4 (d, Jc-p = 6.7 Hz), 44.0, 32.2, 29.20, 29.16, 29.1, 28.9, 28.3, 26.8, 22.5, 21.4 (d, Jc-p = 142.8 Hz). ¹ H NMR (CDCl ₃ , 400 MHz), δ8.49 (d, J = 8.7 Hz, 2H), 6.76 (brm, 1H), 6.18 (d, J = 8.7 Hz, 2H), 5.72 (td, J = 6.6, 15.6 Hz, 1H), 5.45 (dd, J = 15.3 Hz, 1H), 4.92 (brd, J = 8.5 Hz, 1H), 4.15 (m, 1H), 3.74 (d, J = 11.0 Hz, 6H) , 3.63 (m, 1H), 3.50 (td, J = 6.4, 6.4 Hz, 2H), 2.04 (dt, J = 6.6, 6.9 Hz, 2H), 1.60-1.92 (m, 8H), 1.44 (s, 9H ), 1.29-1.48 (m, 12H).
¹³ C NMR (CDCl ₃ , 100 MHz), δ156.4, 144.3, 144.09, 143.95, 136.6, 134.1, 228.5, 123.6, 98.5, 79.7, 75.1, 55.7 (d, Jc-p = 13.4 Hz), 52.4 (d , Jc-p = 6.7 Hz), 44.0, 32.2, 29.20, 29.16, 29.1, 28.9, 28.3, 26.8, 22.5, 21.4 (d, Jc-p = 142.8 Hz).

（実施例７）

(Example 7)

[(3S,4R,5E)-3-tert-ブトキシアルボニルアミノ-4-ヒドロキシ-15-(7-ニトロベンゾ[1,2,5]オキサジアゾール-4-イルアミノ)-ベンタデカ-5-エニル]-ホスホン酸ジメチルエステル(428 mg, 0.682 mmol)のDMF(6.8 ml)溶液に室温でイミダゾール(139 mg, 2.05 mmol)、クロロトリエチルシラン(0.173 ml, 1.02 mmol)を順次加え、１０分間撹拌した。反応混合物に飽和炭酸水素ナトリウムを加え、ジエチルエーテルで抽出した。有機層を飽和食塩水で洗浄し、無水硫酸マグネシウムで乾燥後、減圧濃縮を行った。残渣をシリカゲルクロマトグラフィー（トリエチルアミン3％を含むヘキサンに33〜67％の酢酸エチルを溶かしたもの）により精製し、[(3S,4R,5E)-3-tert-ブトキシアルボニルアミノ-4-トリエチルシロキシ-15-(7-ニトロベンゾ[1,2,5]オキサジアゾール-4-イルアミノ)-ベンタデカ-5-エニル]-ホスホン酸ジメチルエステル(407 mg, 80% yield)を得た。[(3S,4R,5E)-3-tert-ブトキシアルボニルアミノ-4-トリエチルシロキシ-15-(7-ニトロベンゾ[1,2,5]オキサジアゾール-4-イルアミノ)-ベンタデカ-5-エニル]-ホスホン酸ジメチルエステルの¹H NMR、¹³C NMRデータを以下に示す。 [(3S, 4R, 5E) -3-tert-butoxycarbonylamino-4-hydroxy-15- (7-nitrobenzo [1,2,5] oxadiazol-4-ylamino) -bentadeca-5-enyl] -Imidazole (139 mg, 2.05 mmol) and chlorotriethylsilane (0.173 ml, 1.02 mmol) were sequentially added to a DMF (6.8 ml) solution of phosphonic acid dimethyl ester (428 mg, 0.682 mmol) at room temperature and stirred for 10 minutes. Saturated sodium hydrogen carbonate was added to the reaction mixture, and the mixture was extracted with diethyl ether. The organic layer was washed with saturated brine, dried over anhydrous magnesium sulfate, and concentrated under reduced pressure. The residue was purified by silica gel chromatography (33-67% ethyl acetate in hexane containing 3% triethylamine) and [(3S, 4R, 5E) -3-tert-butoxycarbonylamino-4-triethyl Siloxy-15- (7-nitrobenzo [1,2,5] oxadiazol-4-ylamino) -bentadeca-5-enyl] -phosphonic acid dimethyl ester (407 mg, 80% yield) was obtained. [(3S, 4R, 5E) -3-tert-butoxycarbonylamino-4-triethylsiloxy-15- (7-nitrobenzo [1,2,5] oxadiazol-4-ylamino) -bentadeca-5-enyl The ¹ H NMR and ¹³ C NMR data of] -phosphonic acid dimethyl ester are shown below.

¹H NMR (CDCl₃, 400 MHz), δ8.49 (d, J = 8.7 Hz, 2H), 6.81 (brm, 1H), 6.18 (d, J = 8.7 Hz, 2H), 5.63 (td, J = 6.9, 15.4 Hz, 1H), 5.38 (dd, J = 6.9, 15.4 Hz, 1H), 4.66 (brd, J = 8.7 Hz, 2H), 4.17 (m, 1H), 3.74 (d, J = 10.7 Hz, 3H), 3.73 (d, J = 10.8 Hz, 3H), 3.46 - 3.51 (m, 3H), 2.01 (m, 2H), 1.57 - 1.92 (m, 10H), 1.43 (s, 9H), 1.29 - 1.51 (m, 10H), 0.93 (t, J = 7.8 Hz, 9H), 0.57 (q, J = 7.8 Hz, 6H),
¹³C NMR (CDCl₃, 100 MHz), δ155.9, 144.4, 144.2, 144.1, 136.7, 133.1, 129.9, 123.5, 98.4, 79.4, 75.6, 56.3 (d, J_C-P= 16.3 Hz), 52.4 (d, J_C-P= 6.7 Hz), 44.2, 32.1, 29.4, 29.3, 29.2, 29.03, 28.96, 28.4, 27.0, 21.5 (d, J_C-P= 141.8 Hz), 21.5, 6.88, 4.96. ¹ H NMR (CDCl ₃ , 400 MHz), δ8.49 (d, J = 8.7 Hz, 2H), 6.81 (brm, 1H), 6.18 (d, J = 8.7 Hz, 2H), 5.63 (td, J = 6.9, 15.4 Hz, 1H), 5.38 (dd, J = 6.9, 15.4 Hz, 1H), 4.66 (brd, J = 8.7 Hz, 2H), 4.17 (m, 1H), 3.74 (d, J = 10.7 Hz, 3H), 3.73 (d, J = 10.8 Hz, 3H), 3.46-3.51 (m, 3H), 2.01 (m, 2H), 1.57-1.92 (m, 10H), 1.43 (s, 9H), 1.29-1.51 (m, 10H), 0.93 (t, J = 7.8 Hz, 9H), 0.57 (q, J = 7.8 Hz, 6H),
¹³ C NMR (CDCl ₃ , 100 MHz), δ155.9, 144.4, 144.2, 144.1, 136.7, 133.1, 129.9, 123.5, 98.4, 79.4, 75.6, 56.3 (d, J _CP = 16.3 Hz), 52.4 (d, (J _CP = 6.7 Hz), 44.2, 32.1, 29.4, 29.3, 29.2, 29.03, 28.96, 28.4, 27.0, 21.5 (d, J _CP = 141.8 Hz), 21.5, 6.88, 4.96.

（実施例８）

(Example 8)

[(3S,4R,5E)-3-tert-ブトキシアルボニルアミノ-4-トリエチルシロキシ-15-(7-ニトロベンゾ[1,2,5]オキサジアゾール-4-イルアミノ)-ベンタデカ-5-エニル]-ホスホン酸ジメチルエステル(407 mg, 0.549 mmol)の塩化メチレン(5.5 ml)溶液に室温でブロモトリメチルシラン(0.72 ml, 5.49 mmol)を加え、５０分間撹拌した。反応混合物を減圧濃縮し、残渣にメタノール(5.5 ml)を加え、３０分間撹拌した。反応混合物を減圧濃縮し、残渣を逆相シリカゲルクロマトグラフィー（トリフルオロ酢酸１％を含む水に25〜50％のメタノールを溶かしたもの）により粗精製し、[(3S,4R,5E)-3-アミノ-4-ヒドロキシ-15-(7-ニトロベンゾ[1,2,5]オキサジアゾール-4-イルアミノ)-ベンタデカ-5-エニル]-ホスホン酸(289 mg, 85% yield)を得た。[(3S,4R,5E)-3-アミノ-4-ヒドロキシ-15-(7-ニトロベンゾ[1,2,5]オキサジアゾール-4-イルアミノ)-ベンタデカ-5-エニル]-ホスホン酸の¹H NMR、¹³C NMRデータを以下に示す。 [(3S, 4R, 5E) -3-tert-butoxycarbonylamino-4-triethylsiloxy-15- (7-nitrobenzo [1,2,5] oxadiazol-4-ylamino) -bentadeca-5-enyl Bromotrimethylsilane (0.72 ml, 5.49 mmol) was added to a methylene chloride (5.5 ml) solution of] -phosphonic acid dimethyl ester (407 mg, 0.549 mmol) at room temperature and stirred for 50 minutes. The reaction mixture was concentrated under reduced pressure, methanol (5.5 ml) was added to the residue, and the mixture was stirred for 30 min. The reaction mixture was concentrated under reduced pressure, and the residue was roughly purified by reversed-phase silica gel chromatography (25 to 50% methanol in water containing 1% trifluoroacetic acid) and [(3S, 4R, 5E) -3 -Amino-4-hydroxy-15- (7-nitrobenzo [1,2,5] oxadiazol-4-ylamino) -bentadeca-5-enyl] -phosphonic acid (289 mg, 85% yield) was obtained. [(3S, 4R, 5E) -3- amino-4-hydroxy-15- (7-nitrobenzo [1,2,5] oxadiazol-4-ylamino) - Bentadeka-5-enyl] - ¹ phosphonate 1 H NMR and ¹³ C NMR data are shown below.

¹H NMR (CD₃OD, 400 MHz), δ8.42 (d, J = 8.5 Hz, 2H), 6.26 (d, J = 8.9 Hz, 2H), 5.86 (td, J = 6.6, 15.3 Hz, 1H), 5.48 (dd, J = 6.6, 15.4 Hz, 1H), 4.30 (dd, J = 4.3, 5.7 Hz, 1H), 3.48 (m, 2H), 3.30 (m, 1H), 2.07 (td, J = 6.8, 6.8 Hz, 2H), 1.81 - 2.06 (m, 4H), 1.75 (tt, J = 7.3, 7.3 Hz, 2H), 1.28 - 1.46 (m, 12H).
¹³C NMR (CD₃OD, 100 MHz), δ146.5, 145.6, 145.3, 138.5, 136.9, 122.5, 99.6, 72.3, 57.5 (d, J_C-P= 15.3 Hz), 44.8, 33.4, 30.52, 30.45, 30.33, 30.30, 30.1, 29.2, 28.0, 24.7 (d, J_C-P= 138.9 Hz), 23.2. ¹ H NMR (CD ₃ OD, 400 MHz), δ8.42 (d, J = 8.5 Hz, 2H), 6.26 (d, J = 8.9 Hz, 2H), 5.86 (td, J = 6.6, 15.3 Hz, 1H ), 5.48 (dd, J = 6.6, 15.4 Hz, 1H), 4.30 (dd, J = 4.3, 5.7 Hz, 1H), 3.48 (m, 2H), 3.30 (m, 1H), 2.07 (td, J = 6.8, 6.8 Hz, 2H), 1.81-2.06 (m, 4H), 1.75 (tt, J = 7.3, 7.3 Hz, 2H), 1.28-1.46 (m, 12H).
¹³ C NMR (CD ₃ OD, 100 MHz), δ146.5, 145.6, 145.3, 138.5, 136.9, 122.5, 99.6, 72.3, 57.5 (d, J _CP = 15.3 Hz), 44.8, 33.4, 30.52, 30.45, 30.33 , 30.30, 30.1, 29.2, 28.0, 24.7 (d, J _CP = 138.9 Hz), 23.2.

Claims

The following general formula (I)

Wherein R ¹ and R ² are a hydrogen atom or a lower alkyl group, Q ¹ and Q ² are a hydrogen atom or an amino group protecting group, Q ³ is a hydrogen atom or a hydroxyl protecting group, Is an alkylene group having 1 to 15 carbon atoms and Z is a fluorescent labeling group, Q ¹ and Q ² , Q ² and Q ^3, or Q ¹ and Q ³ may be combined to form a ring. A fluorescently labeled sphingosine 1-methylene phosphate substitute or an acceptable salt thereof.