JP7296636B2 - Alkoxyphenyl derivative, protected nucleoside and protected nucleotide, method for producing oligonucleotide, and method for removing substituent - Google Patents

Description

The present invention provides an alkoxyphenyl derivative that enables the synthesis of oligonucleotides by a liquid phase synthesis method, a nucleoside-protected body and a nucleotide-protected body bound thereto, a method for producing an oligonucleotide that can be easily constructed using the same, and The present invention relates to a method for selectively removing an alkoxyphenyl derivative portion of a protected nucleoside or nucleotide to which an alkoxyphenyl derivative is bound.

In recent years, oligonucleotides can generally be synthesized by sequentially linking nucleotides as starting materials, but in the case of synthesizing oligonucleotides of about 20mer or more, building blocks of 2 to 3mer nucleotides are prepared in advance. A blocker synthesis method that obtains a product of a desired chain length by repeating the ligation of this prepared product, and a unit coupling (fragment condensation) method that obtains an oligonucleotide by ligation between oligonucleotides of about 10 bases or more. ) are also used (for example, Non-Patent Documents 1 to 3). In solid-phase synthesis of oligonucleotides using the phosphoramidite method, a large excess of nucleoside phosphoramidite compound and tetrazole compound must be used in order to increase the yield of the target oligonucleotide. There is a problem. In addition, in the solid-phase synthesis method,
There are restrictions on scale-up in terms of equipment, etc., and it is difficult to check the progress of the reaction in the middle and to analyze the structure of the intermediate.

On the other hand, conventional liquid-phase synthesis methods require complicated purification procedures in each step of deprotection, coupling, and oxidation of nucleotides. was difficult. For this reason, for example, Patent Documents 1 and 2 propose liquid-phase synthesis using a pseudo-solid-phase protecting group, but for example, it takes time to remove the pseudo-solid-phase protecting group, and furthermore, However, since it is difficult to selectively remove only the pseudo solid-phase protecting group from an oligonucleotide having a pseudo solid-phase protecting group, the nucleobases and phosphate groups that are precursors of building blocks and fragments are protected. It has also been difficult to synthesize such nucleotides (hereinafter sometimes referred to as "protected nucleotides" in this specification). In addition, in Non-Patent Document 4, the pseudo-solid-phase protective group is removed by catalytic reduction using a Pd catalyst. It takes a long time. In addition, the pseudo-solid-phase protecting group has not been removed under general deprotection conditions for protecting groups of nucleotides, and there is a problem in its versatility.

Patent No. 5548852 WO2012/157723

S. L. Beaucage, D.; E. Bergstorm, G.; D. Glick, R. A. Jones; Current Protocols in Nucleic Acid Chemistry; John Wiley & Sons (2000) Mitsuo Sekine, Retsu Saito, eds., "Genome Chemistry - A Scientific Approach Utilizing Artificial Nucleic Acids", Kodansha Scientific (2003) C. -H. Chen, et al. , Aust. J. Chem. , 2010, 63, 227-235. Y. Matsuno, et al. , Org. Lett. , 2016, 18, 800-803.

In light of such circumstances, the present invention provides a highly versatile alkoxyphenyl derivative that enables not only the synthesis of oligonucleotides but also the easy and rapid synthesis of protected oligonucleotides, which has been difficult in the past. aim.

In addition, the present invention provides not only the synthesis of oligonucleotides, but also the highly versatile alkoxyphenyl derivative that enables simple and rapid synthesis of protected oligonucleotides, which has been difficult in the past (hereinafter referred to as In some cases, it is also referred to as "tagging".) Nucleoside protected body and nucleotide protected body (hereinafter, in this specification, respectively, "Tag
It is sometimes referred to as "protected nucleoside" and "protected tagged nucleotide". ) is intended to provide

Furthermore, the present invention provides an oligonucleotide production method (synthesis method) that is more versatile than conventional ones, using the above alkoxyphenyl derivative, or the above tagged nucleoside protected form or tagged nucleotide protected form. aim.

Another object of the present invention is to provide a method for selectively removing the Tag portion in a protected nucleoside tagged or protected nucleotide tagged. In the present invention, "Tag portion" refers to a portion derived from the alkoxyphenyl derivative in the protected tagged nucleoside or protected nucleotide.

The inventors of the present invention have made intensive studies to solve the above problems, and as a result, have succeeded in creating the novel alkoxyphenyl compounds and derivatives thereof shown below, and found that the above objects can be achieved by the above compounds, thereby completing the present invention. reached.

（式中、
Ｒは、それぞれ独立して、置換されていてもよい、炭素数１０～４０のアルキル基を表
す。
ｍは、１～５の整数を表す。ｍが２以上の場合には、複数存在するＲＯは同一、または
、異なっていてもよい。
Ｘは、Ｏ、Ｓ、ＮＨ、または、ＮＲ^Ｎを表す。
ｎは、１～４の整数を表す。
Ｒ^Ｎは、置換されていてもよい、炭素数１～６のアルキル基を表す。） That is, the compounds and derivatives thereof of the present invention are represented by general formula (1) (hereinafter, sometimes referred to as "compound (1)" in the present specification). Compounds represented by other general formulas are also according to ).

(In the formula,
Each R independently represents an optionally substituted alkyl group having 10 to 40 carbon atoms.
m represents an integer of 1 to 5; When m is 2 or more, multiple ROs may be the same or different.
X represents O, S, NH, or ^NRN .
n represents an integer of 1-4.
R ^N represents an optionally substituted alkyl group having 1 to 6 carbon atoms. )

According to the compound or derivative thereof of the present invention, by having the structure represented by the general formula (1), not only the synthesis of oligonucleotides by the liquid phase synthesis method, but also the protection of oligonucleotides, which has been difficult in the past, can be obtained. Synthesis becomes possible. More specifically, the compound of the present invention or a derivative thereof (alkoxyphenyl derivative) can obtain protected nucleoside tagged and protected nucleotide tagged via binding reaction with nucleoside or nucleotide. Then, the obtained tagged nucleoside protected body and tagged nucleotide protected body are easily separated from auxiliary materials such as amidite monomers by addition of a polar solvent and filtering operation etc., because the Tag portion has the above structure. Furthermore, it is possible to selectively and rapidly remove only the Tag portion, and as a result, it is possible to synthesize (manufacture) protected oligonucleotides, which has been difficult in the past, as well as to synthesize oligonucleotides. Become.

Derivatives of the above compounds broadly include those obtained by partially substituting the chemical structure of the above compounds as long as they do not impair the effects of the present invention.
COOH) groups in which the OH group portion is replaced with a substituent. Examples of the above-mentioned substituents include substituents having high leaving ability so as to facilitate bond formation with nucleosides or nucleotides. An ester structure and the like can be mentioned. Moreover, you may use a well-known leaving group suitably as said substituent.

（式中、
Ｒ^１、Ｒ^２、および、Ｒ^３は、それぞれ独立して、置換されていてもよい、炭素数１０
～４０のアルキル基を表す。
Ｘは、Ｏ、Ｓ、ＮＨ、または、ＮＲ^Ｎを表す。
ｎは、１～４の整数を表す。
Ｒ^Ｎは、置換されていてもよい、炭素数１～６のアルキル基を表す。） Moreover, the compound or derivative thereof of the present invention is preferably a compound represented by the general formula (2) or a derivative thereof.

(In the formula,
R ¹ , R ² and R ³ each independently have 10 carbon atoms and may be substituted.
represents an alkyl group of ∼40.
X represents O, S, NH, or ^NRN .
n represents an integer of 1-4.
R ^N represents an optionally substituted alkyl group having 1 to 6 carbon atoms. )

（式中、
Ｒは、それぞれ独立して、置換されていてもよい、炭素数１０～４０のアルキル基を表
す。
ｍは、１～５の整数を表す。ｍが２以上の場合には、複数存在するＲＯは、同一、また
は、異なっていてもよい。
Ｘは、Ｏ、Ｓ、ＮＨ、または、ＮＲ^Ｎを表す。
ｎは、１～４の整数を表す。
Ｒ^Ｎは、置換されていてもよい、炭素数１～６のアルキル基を表す。
Ｙは、下記一般式（４）、または、一般式（５）を表す。）

（式中、
Ｂａｓｅは、修飾されていてもよい核酸塩基を表す。Ｂａｓｅが２以上存在する場合に
は、複数存在するＢａｓｅは同一、または、異なっていてもよい。
Ｂは、水素原子、保護されていてもよい水酸基、または、ハロゲンを表す。Ｂが２以上
存在する場合には、複数存在するＢは、同一、または、異なっていてもよい。
Ｗは、水素原子、または、保護基を表す。
Ｒ^４は、電子求引性基を有する、炭素数１～４のアルキル基を表す。Ｒ^４が２以上存在
する場合には、複数存在するＲ^４は同一、または、異なっていてもよい。
Ｚは、Ｏ、または、Ｓを表す。Ｚが２以上存在する場合には、複数存在するＺは、同一
、または、異なっていてもよい。
ｙ１は、０以上の任意の整数を表す。
＊は、上記一般式（３）における結合箇所を表す。）

（式中、
Ｂａｓｅは、修飾されていてもよい核酸塩基を表す。Ｂａｓｅが２以上存在する場合に
は、複数存在するＢａｓｅは同一、または、異なっていてもよい。
Ｑは、水素原子、または、保護基を表す。
Ｖは、炭素数１～６のアルコキシ基、ジ（Ｃ_１－６アルキル）アミノ基、または、４位
窒素原子が保護基で保護され、さらに置換されていてもよいピペラジノ基を表す。Ｖが２
以上存在する場合には、複数存在するＶは同一、または、異なっていてもよい。
ｙ２は、０以上の任意の整数を表す。
＊は、上記一般式（３）における結合箇所を表す。） Moreover, the compound or its derivative of this invention WHEREIN: The compound represented by General formula (3) or its derivative may be sufficient.

(In the formula,
Each R independently represents an optionally substituted alkyl group having 10 to 40 carbon atoms.
m represents an integer of 1 to 5; When m is 2 or more, multiple ROs may be the same or different.
X represents O, S, NH, or ^NRN .
n represents an integer of 1-4.
R ^N represents an optionally substituted alkyl group having 1 to 6 carbon atoms.
Y represents the following general formula (4) or general formula (5). )

(In the formula,
Base represents an optionally modified nucleobase. When two or more Bases exist, the plurality of Bases may be the same or different.
B represents a hydrogen atom, an optionally protected hydroxyl group, or a halogen. When two or more B are present, the plural B may be the same or different.
W represents a hydrogen atom or a protecting group.
R ⁴ represents a C 1-4 alkyl group having an electron-withdrawing group. When two or more R ⁴ are present, the plurality of R ⁴ may be the same or different.
Z represents O or S; When there are two or more Z's, the multiple Z's may be the same or different.
y1 represents any integer of 0 or more.
* represents a bonding site in the above general formula (3). )

(In the formula,
Base represents an optionally modified nucleobase. When two or more Bases exist, the plurality of Bases may be the same or different.
Q represents a hydrogen atom or a protecting group.
V represents an alkoxy group having 1 to 6 carbon atoms, a di(C _1-6 alkyl)amino group, or an optionally substituted piperazino group whose 4-position nitrogen atom is protected by a protecting group. V is 2
When there are more than one V, the plurality of V may be the same or different.
y2 represents any integer of 0 or more.
* represents a bonding site in the general formula (3). )

When it has the above structure, it becomes the above tagged nucleoside protected body and tagged nucleotide protected body. , It can be easily separated from auxiliary materials such as amidite monomers by a filtration operation, etc., and it is possible to selectively and quickly remove only the Tag portion. It becomes possible to synthesize protected oligonucleotides, which has been difficult.

In addition, the nucleoside in the present invention generally represents a glycosidic bond between a purine or pyrimidine base and a sugar, but as long as it does not impair the action and effect of the present invention, its analogue or partial substitution of the general structure widely included. For example, base moieties are not necessarily purine or pyrimidine structures (e.g., modified bases, etc.), and sugar moieties are not D-ribose or deoxy-D-ribose (e.g., general formulas (5) and (8) and morpholino-type morpholino nucleosides, such as shown in the above.

In addition, the nucleotide in the present invention generally represents a phosphate ester of the above nucleoside in which a purine or pyrimidine base and a sugar are glycoside-bonded. Those with partial replacement of the structure are widely included.
For example, those in which the base moiety is not necessarily a purine or pyrimidine structure (e.g., modified bases, etc.), those in which the sugar moiety is not D-ribose or deoxy-D-ribose (e.g., morpholino-type morpholino nucleotides), Phosphorothioate in which one O atom of the phosphate ester moiety is replaced with an S atom can be mentioned.

（式中、
Ｒ^１、Ｒ^２、および、Ｒ^３は、それぞれ独立して、置換されていてもよい、炭素数１０
～４０のアルキル基を表す。
Ｘは、Ｏ、Ｓ、ＮＨ、または、ＮＲ^Ｎを表す。
ｎは、１～４の整数を表す。
Ｒ^Ｎは、置換されていてもよい、炭素数１～６のアルキル基を表す。
Ｙは、下記一般式（４）、または、一般式（５）を表す。）

（式中、
Ｂａｓｅは、修飾されていてもよい核酸塩基を表す。Ｂａｓｅが２以上存在する場合に
は、複数存在するＢａｓｅは同一、または、異なっていてもよい。
Ｂは、水素原子、保護されていてもよい水酸基、または、ハロゲンを表す。Ｂが２以上
存在する場合には、複数存在するＢは、同一、または、異なっていてもよい。
Ｗは、水素原子、または、保護基を表す。
Ｒ^４は、電子求引性基を有する、炭素数１～４のアルキル基を表す。Ｒ^４が２以上存在
する場合には、複数存在するＲ^４は同一、または、異なっていてもよい。
Ｚは、Ｏ、または、Ｓを表す。Ｚが２以上存在する場合には、複数存在するＺは、同一
、または、異なっていてもよい。
ｙ１は、０以上の任意の整数を表す。
＊は、上記一般式（６）における結合箇所を表す。）

（式中、
Ｂａｓｅは、修飾されていてもよい核酸塩基を表す。Ｂａｓｅが２以上存在する場合に
は、複数存在するＢａｓｅは同一、または、異なっていてもよい。
Ｑは、水素原子、または、保護基を表す。
Ｖは、炭素数１～６のアルコキシ基、ジ（Ｃ_１－６アルキル）アミノ基、または、４位
窒素原子が保護基で保護され、さらに置換されていてもよいピペラジノ基を表す。Ｖが２
以上存在する場合には、複数存在するＶは同一、または、異なっていてもよい。
ｙ２は、０以上の任意の整数を表す。
＊は、上記一般式（６）における結合箇所を表す。） Moreover, the compound or derivative thereof of the present invention is preferably a compound represented by the general formula (6) or a derivative thereof.

(In the formula,
R ¹ , R ² and R ³ each independently have 10 carbon atoms and may be substituted.
represents an alkyl group of ∼40.
X represents O, S, NH, or ^NRN .
n represents an integer of 1-4.
R ^N represents an optionally substituted alkyl group having 1 to 6 carbon atoms.
Y represents the following general formula (4) or general formula (5). )

(In the formula,
Base represents an optionally modified nucleobase. When two or more Bases exist, the plurality of Bases may be the same or different.
B represents a hydrogen atom, an optionally protected hydroxyl group, or a halogen. When two or more B are present, the plural B may be the same or different.
W represents a hydrogen atom or a protecting group.
R ⁴ represents a C 1-4 alkyl group having an electron-withdrawing group. When two or more R ⁴ are present, the plurality of R ⁴ may be the same or different.
Z represents O or S; When there are two or more Z's, the multiple Z's may be the same or different.
y1 represents any integer of 0 or more.
* represents a bonding site in the general formula (6). )

(In the formula,
Base represents an optionally modified nucleobase. When two or more Bases exist, the plurality of Bases may be the same or different.
Q represents a hydrogen atom or a protecting group.
V represents an alkoxy group having 1 to 6 carbon atoms, a di(C _1-6 alkyl)amino group, or an optionally substituted piperazino group whose 4-position nitrogen atom is protected by a protecting group. V is 2
When there are more than one V, the plurality of V may be the same or different.
y2 represents any integer of 0 or more.
* represents a bonding site in the general formula (6). )

Further, in the compound or derivative thereof of the present invention, W is a hydrogen atom, a trityl group, a di(
C _1-6 alkoxy)trityl group, mono (C _1-18 alkoxy)trityl group, 9-(9
-phenyl)xanthenyl group or 9-(9-phenyl)thioxanthenyl group is preferred. When W is any of the above, oligonucleotides can be synthesized more easily.

Moreover, in the compound or derivative thereof of the present invention, ^R4 is preferably an ethyl group having an electron-withdrawing group at the 2-position.

Further, in the compound or derivative thereof of the present invention, Q is a hydrogen atom, a trityl group, a di(
C _1-6 alkoxy)trityl group, mono (C _1-18 alkoxy)trityl group, 9-(9
-phenyl)xanthenyl group or 9-(9-phenyl)thioxanthenyl group is preferred. When Q is any of the above, oligonucleotides can be synthesized more easily.

Further, in the compound or derivative thereof of the present invention, y1 or y2 is 1 to 100
may be

In addition, in the compound or derivative thereof of the present invention, y1 or y2 may be 1-30.

In addition, y1 or y2 may be 0 in the compound or derivative thereof of the present invention.

Moreover, n is preferably 1 or 2 in the compound or derivative thereof of the present invention.

In the compound or derivative thereof of the present invention, m is preferably an integer of 2-4.

（式中、
Ｒは、それぞれ独立して、置換されていてもよい、炭素数１０～４０のアルキル基を表
す。
ｍは、１～５の整数を表す。ｍが２以上の場合には、複数存在するＲＯは、同一、また
は、異なっていてもよい。
Ｘは、Ｏ、Ｓ、ＮＨ、または、ＮＲ^Ｎを表す。
ｎは、１～４の整数を表す。
Ｒ^Ｎは、置換されていてもよい、炭素数１～６のアルキル基を表す。
Ｙは、下記一般式（４）、または、一般式（５）を表す。）

（式中、
Ｂａｓｅは、修飾されていてもよい核酸塩基を表す。Ｂａｓｅが２以上存在する場合に
は、複数存在するＢａｓｅは同一、または、異なっていてもよい。
Ｂは、水素原子、保護されていてもよい水酸基、または、ハロゲンを表す。Ｂが２以上
存在する場合には、複数存在するＢは、同一、または、異なっていてもよい。
Ｗは、水素原子、または、保護基を表す。
Ｒ^４は、電子求引性基を有する、炭素数１～４のアルキル基を表す。Ｒ^４が２以上存在
する場合には、複数存在するＲ^４は同一、または、異なっていてもよい。
Ｚは、Ｏ、または、Ｓを表す。Ｚが２以上存在する場合には、複数存在するＺは、同一
、または、異なっていてもよい。
ｙ１は、０以上の任意の整数を表す。
＊は、上記一般式（３）における結合箇所を表す。）

（式中、
Ｂａｓｅは、修飾されていてもよい核酸塩基を表す。Ｂａｓｅが２以上存在する場合に
は、複数存在するＢａｓｅは同一、または、異なっていてもよい。
Ｑは、水素原子、または、保護基を表す。
Ｖは、炭素数１～６のアルコキシ基、ジ（Ｃ_１－６アルキル）アミノ基、または、４位
窒素原子が保護基で保護され、さらに置換されていてもよいピペラジノ基を表す。Ｖが２
以上存在する場合には、複数存在するＶは同一、または、異なっていてもよい。
ｙ２は、０以上の任意の整数を表す。
＊は、上記一般式（３）における結合箇所を表す。）

（式中、
Ｂａｓｅは、修飾されていてもよい核酸塩基を表す。Ｂａｓｅが２以上存在する場合に
は、複数存在するＢａｓｅは同一、または、異なっていてもよい。
Ｂは、水素原子、保護されていてもよい水酸基、または、ハロゲンを表す。Ｂが２以上
存在する場合には、複数存在するＢは、同一、または、異なっていてもよい。
Ｗは、水素原子、または、保護基を表す。
Ｒ^４は、電子求引性基を有する、炭素数１～４のアルキル基を表す。Ｒ^４が２以上存在
する場合には、複数存在するＲ^４は同一、または、異なっていてもよい。
Ｚは、Ｏ、または、Ｓを表す。Ｚが２以上存在する場合には、複数存在するＺは、同一
、または、異なっていてもよい。
ｙ１は、０以上の任意の整数を表す。）

（式中、
Ｂａｓｅは、修飾されていてもよい核酸塩基を表す。Ｂａｓｅが２以上存在する場合に
は、複数存在するＢａｓｅは同一、または、異なっていてもよい。
Ｑは、水素原子、または、保護基を表す。
Ｖは、炭素数１～６のアルコキシ基、ジ（Ｃ_１－６アルキル）アミノ基、または、４位
窒素原子が保護基で保護され、さらに置換されていてもよいピペラジノ基を表す。Ｖが２
以上存在する場合には、複数存在するＶは同一、または、異なっていてもよい。
ｙ２は、０以上の任意の整数を表す。） On the other hand, the production method of the present invention includes step (1) of obtaining a compound represented by general formula (7) or (8) by reducing the compound represented by general formula (3) or a derivative thereof.

(In the formula,
Each R independently represents an optionally substituted alkyl group having 10 to 40 carbon atoms.
m represents an integer of 1 to 5; When m is 2 or more, multiple ROs may be the same or different.
X represents O, S, NH, or ^NRN .
n represents an integer of 1-4.
R ^N represents an optionally substituted alkyl group having 1 to 6 carbon atoms.
Y represents the following general formula (4) or general formula (5). )

(In the formula,
Base represents an optionally modified nucleobase. When two or more Bases exist, the plurality of Bases may be the same or different.
B represents a hydrogen atom, an optionally protected hydroxyl group, or a halogen. When two or more B are present, the plural B may be the same or different.
W represents a hydrogen atom or a protecting group.
R ⁴ represents a C 1-4 alkyl group having an electron-withdrawing group. When two or more R ⁴ are present, the plurality of R ⁴ may be the same or different.
Z represents O or S; When there are two or more Z's, the multiple Z's may be the same or different.
y1 represents any integer of 0 or more.
* represents a bonding site in the above general formula (3). )

(In the formula,
Base represents an optionally modified nucleobase. When two or more Bases exist, the plurality of Bases may be the same or different.
Q represents a hydrogen atom or a protecting group.
V represents an alkoxy group having 1 to 6 carbon atoms, a di(C _1-6 alkyl)amino group, or an optionally substituted piperazino group whose 4-position nitrogen atom is protected by a protecting group. V is 2
When there are more than one V, the plurality of V may be the same or different.
y2 represents any integer of 0 or more.
* represents a bonding site in the above general formula (3). )

(In the formula,
Base represents an optionally modified nucleobase. When two or more Bases exist, the plurality of Bases may be the same or different.
B represents a hydrogen atom, an optionally protected hydroxyl group, or a halogen. When two or more B are present, the plural B may be the same or different.
W represents a hydrogen atom or a protecting group.
R ⁴ represents a C 1-4 alkyl group having an electron-withdrawing group. When two or more R ⁴ are present, the plurality of R ⁴ may be the same or different.
Z represents O or S; When there are two or more Z's, the multiple Z's may be the same or different.
y1 represents any integer of 0 or more. )

(In the formula,
Base represents an optionally modified nucleobase. When two or more Bases exist, the plurality of Bases may be the same or different.
Q represents a hydrogen atom or a protecting group.
V represents an alkoxy group having 1 to 6 carbon atoms, a di(C _1-6 alkyl)amino group, or an optionally substituted piperazino group whose 4-position nitrogen atom is protected by a protecting group. V is 2
When there are more than one V, the plurality of V may be the same or different.
y2 represents any integer of 0 or more. )

According to the production method of the present invention, since the tagged nucleoside protected body and the tagged nucleotide protected body are used, especially in liquid phase synthesis (including pseudo-liquid phase synthesis), secondary raw materials such as amidite monomers can be obtained by adding a polar solvent. It can be easily separated by filtration operation etc., and it is possible to selectively and quickly remove only the Tag part,
It enables not only the synthesis of oligonucleotides but also the synthesis of protected oligonucleotides, which has been difficult in the past. In addition, for example, compared with the proposal of Patent Document 2, the production method of the present invention enables removal of substituents (elimination reaction of Tag moiety) under milder conditions (for example, 35° C. in ammonia water), and , making it possible to manufacture in a shorter time.

（式中、
Ｒ^１、Ｒ^２、および、Ｒ^３は、それぞれ独立して、置換されていてもよい、炭素数１０
～４０のアルキル基を表す。
Ｘは、Ｏ、Ｓ、ＮＨ、または、ＮＲ^Ｎを表す。
ｎは、１～４の整数を表す。
Ｒ^Ｎは、置換されていてもよい、炭素数１～６のアルキル基を表す。
Ｙは、下記一般式（４）、または、一般式（５）を表す。）

（式中、
Ｂａｓｅは、修飾されていてもよい核酸塩基を表す。Ｂａｓｅが２以上存在する場合に
は、複数存在するＢａｓｅは同一、または、異なっていてもよい。
Ｂは、水素原子、保護されていてもよい水酸基、または、ハロゲンを表す。Ｂが２以上
存在する場合には、複数存在するＢは、同一、または、異なっていてもよい。
Ｗは、水素原子、または、保護基を表す。
Ｒ^４は、電子求引性基を有する、炭素数１～４のアルキル基を表す。Ｒ^４が２以上存在
する場合には、複数存在するＲ^４は同一、または、異なっていてもよい。
Ｚは、Ｏ、または、Ｓを表す。Ｚが２以上存在する場合には、複数存在するＺは、同一
、または、異なっていてもよい。
ｙ１は、０以上の任意の整数を表す。
＊は、上記一般式（６）における結合箇所を表す。）

（式中、
Ｂａｓｅは、修飾されていてもよい核酸塩基を表す。Ｂａｓｅが２以上存在する場合に
は、複数存在するＢａｓｅは同一、または、異なっていてもよい。
Ｑは、水素原子、または、保護基を表す。
Ｖは、炭素数１～６のアルコキシ基、ジ（Ｃ_１－６アルキル）アミノ基、または、４位
窒素原子が保護基で保護され、さらに置換されていてもよいピペラジノ基を表す。Ｖが２
以上存在する場合には、複数存在するＶは同一、または、異なっていてもよい。
ｙ２は、０以上の任意の整数を表す。
＊は、上記一般式（６）における結合箇所を表す。）

（式中、
Ｂａｓｅは、修飾されていてもよい核酸塩基を表す。Ｂａｓｅが２以上存在する場合に
は、複数存在するＢａｓｅは同一、または、異なっていてもよい。
Ｂは、水素原子、保護されていてもよい水酸基、または、ハロゲンを表す。Ｂが２以上
存在する場合には、複数存在するＢは、同一、または、異なっていてもよい。
Ｗは、水素原子、または、保護基を表す。
Ｒ^４は、電子求引性基を有する、炭素数１～４のアルキル基を表す。Ｒ^４が２以上存在
する場合には、複数存在するＲ^４は同一、または、異なっていてもよい。
Ｚは、Ｏ、または、Ｓを表す。Ｚが２以上存在する場合には、複数存在するＺは、同一
、または、異なっていてもよい。
ｙ１は、０以上の任意の整数を表す。）

（式中、
Ｂａｓｅは、修飾されていてもよい核酸塩基を表す。Ｂａｓｅが２以上存在する場合に
は、複数存在するＢａｓｅは同一、または、異なっていてもよい。
Ｑは、水素原子、または、保護基を表す。
Ｖは、炭素数１～６のアルコキシ基、ジ（Ｃ_１－６アルキル）アミノ基、または、４位
窒素原子が保護基で保護され、さらに置換されていてもよいピペラジノ基を表す。Ｖが２
以上存在する場合には、複数存在するＶは同一、または、異なっていてもよい。
ｙ２は、０以上の任意の整数を表す。） Moreover, the method for producing a protected oligonucleotide of the present invention preferably includes the step (1) of reducing the compound represented by the general formula (6) or its derivative.

(In the formula,
R ¹ , R ² and R ³ each independently have 10 carbon atoms and may be substituted.
represents an alkyl group of ∼40.
X represents O, S, NH, or ^NRN .
n represents an integer of 1-4.
R ^N represents an optionally substituted alkyl group having 1 to 6 carbon atoms.
Y represents the following general formula (4) or general formula (5). )

(In the formula,
Base represents an optionally modified nucleobase. When two or more Bases exist, the plurality of Bases may be the same or different.
B represents a hydrogen atom, an optionally protected hydroxyl group, or a halogen. When two or more B are present, the plural B may be the same or different.
W represents a hydrogen atom or a protecting group.
R ⁴ represents a C 1-4 alkyl group having an electron-withdrawing group. When two or more R ⁴ are present, the plurality of R ⁴ may be the same or different.
Z represents O or S; When there are two or more Z's, the multiple Z's may be the same or different.
y1 represents any integer of 0 or more.
* represents a bonding site in the general formula (6). )

(In the formula,
Base represents an optionally modified nucleobase. When two or more Bases exist, the plurality of Bases may be the same or different.
Q represents a hydrogen atom or a protecting group.
V represents an alkoxy group having 1 to 6 carbon atoms, a di(C _1-6 alkyl)amino group, or an optionally substituted piperazino group whose 4-position nitrogen atom is protected by a protecting group. V is 2
When there are more than one V, the plurality of V may be the same or different.
y2 represents any integer of 0 or more.
* represents a bonding site in the general formula (6). )

(In the formula,
Base represents an optionally modified nucleobase. When two or more Bases exist, the plurality of Bases may be the same or different.
B represents a hydrogen atom, an optionally protected hydroxyl group, or a halogen. When two or more B are present, the plural B may be the same or different.
W represents a hydrogen atom or a protecting group.
R ⁴ represents a C 1-4 alkyl group having an electron-withdrawing group. When two or more R ⁴ are present, the plurality of R ⁴ may be the same or different.
Z represents O or S; When there are two or more Z's, the multiple Z's may be the same or different.
y1 represents any integer of 0 or more. )

(In the formula,
Base represents an optionally modified nucleobase. When two or more Bases exist, the plurality of Bases may be the same or different.
Q represents a hydrogen atom or a protecting group.
V represents an alkoxy group having 1 to 6 carbon atoms, a di(C _1-6 alkyl)amino group, or an optionally substituted piperazino group whose 4-position nitrogen atom is protected by a protecting group. V is 2
When there are more than one V, the plurality of V may be the same or different.
y2 represents any integer of 0 or more. )

Moreover, in the production method of the present invention, the reduction treatment preferably uses a boron-containing reducing agent or uses a boron-containing reducing agent and an amine. By having the above structure, the protected oligonucleotide can be synthesized more easily.

Further, in the production method of the present invention, the boron-containing reducing agent is lithium borohydride,
Preferred are sodium borohydride, lithium triethylborohydride, and tetrabutylammonium borohydride. By having the above structure, the protected oligonucleotide can be easily synthesized with higher yield.

Further, in the production method of the present invention, W is a hydrogen atom, a trityl group, a di(C _1-6 alkoxy)trityl group, a mono(C _1-18 alkoxy)trityl group, a 9-(9-phenyl)xanthenyl group, Alternatively, it is preferably a 9-(9-phenyl)thioxanthenyl group. When W is any of the above, protected oligonucleotides can be synthesized more easily.

Moreover, in the production method of the present invention, ^R4 is preferably an ethyl group having an electron-withdrawing group at the 2-position.

Further, in the production method of the present invention, Q is a hydrogen atom, a trityl group, a di(C _1-6 alkoxy)trityl group, a mono(C _1-18 alkoxy)trityl group, a 9-(9-phenyl)xanthenyl group, Alternatively, it is preferably a 9-(9-phenyl)thioxanthenyl group. When Q is any of the above, oligonucleotides can be synthesized more easily.

In the production method of the present invention, y1 or y2 may be 1-100.

Further, in the production method of the present invention, y1 or y2 may be 1-30.

Moreover, in the production method of the present invention, y1 or y2 may be 0.

Moreover, in the production method of the present invention, n is preferably 1 or 2.

Further, in the production method of the present invention, m is preferably an integer of 2-4.

（式中、
Ｒは、それぞれ独立して、置換されていてもよい、炭素数１０～４０のアルキル基を表
す。
ｍは、１～５の整数を表す。ｍが２以上の場合には、複数存在するＲＯは、同一、また
は、異なっていてもよい。
Ｘは、Ｏ、Ｓ、ＮＨ、または、ＮＲ^Ｎを表す。
ｎは、１～４の整数を表す。
Ｒ^Ｎは、置換されていてもよい、炭素数１～６のアルキル基を表す。
Ｙは、下記一般式（４）、または、一般式（５）を表す。）

（式中、
Ｂａｓｅは、修飾されていてもよい核酸塩基を表す。Ｂａｓｅが２以上存在する場合に
は、複数存在するＢａｓｅは同一、または、異なっていてもよい。
Ｂは、水素原子、保護されていてもよい水酸基、または、ハロゲンを表す。Ｂが２以上
存在する場合には、複数存在するＢは、同一、または、異なっていてもよい。
Ｗは、水素原子、または、保護基を表す。
Ｒ^４は、電子求引性基を有する、炭素数１～４のアルキル基を表す。Ｒ^４が２以上存在
する場合には、複数存在するＲ^４は同一、または、異なっていてもよい。
Ｚは、Ｏ、または、Ｓを表す。Ｚが２以上存在する場合には、複数存在するＺは、同一
、または、異なっていてもよい。
ｙ１は、０以上の任意の整数を表す。
＊は、上記一般式（３）における結合箇所を表す。）

（式中、
Ｂａｓｅは、修飾されていてもよい核酸塩基を表す。Ｂａｓｅが２以上存在する場合に
は、複数存在するＢａｓｅは同一、または、異なっていてもよい。
Ｑは、水素原子、または、保護基を表す。
Ｖは、炭素数１～６のアルコキシ基、ジ（Ｃ_１－６アルキル）アミノ基、または、４位
窒素原子が保護基で保護され、さらに置換されていてもよいピペラジノ基を表す。Ｖが２
以上存在する場合には、複数存在するＶは同一、または、異なっていてもよい。
ｙ２は、０以上の任意の整数を表す。
＊は、上記一般式（３）における結合箇所を表す。） On the other hand, the Tag moiety removal method (substituent removal method) of the present invention includes step (1) of reducing the compound represented by general formula (3) or a derivative thereof.

(In the formula,
Each R independently represents an optionally substituted alkyl group having 10 to 40 carbon atoms.
m represents an integer of 1 to 5; When m is 2 or more, multiple ROs may be the same or different.
X represents O, S, NH, or ^NRN .
n represents an integer of 1-4.
R ^N represents an optionally substituted alkyl group having 1 to 6 carbon atoms.
Y represents the following general formula (4) or general formula (5). )

(In the formula,
Base represents an optionally modified nucleobase. When two or more Bases exist, the plurality of Bases may be the same or different.
B represents a hydrogen atom, an optionally protected hydroxyl group, or a halogen. When two or more B are present, the plural B may be the same or different.
W represents a hydrogen atom or a protecting group.
R ⁴ represents a C 1-4 alkyl group having an electron-withdrawing group. When two or more R ⁴ are present, the plurality of R ⁴ may be the same or different.
Z represents O or S; When there are two or more Z's, the multiple Z's may be the same or different.
y1 represents any integer of 0 or more.
* represents a bonding site in the above general formula (3). )

(In the formula,
Base represents an optionally modified nucleobase. When two or more Bases exist, the plurality of Bases may be the same or different.
Q represents a hydrogen atom or a protecting group.
V represents an alkoxy group having 1 to 6 carbon atoms, a di(C _1-6 alkyl)amino group, or an optionally substituted piperazino group whose 4-position nitrogen atom is protected by a protecting group. V is 2
When there are more than one V, the plurality of V may be the same or different.
y2 represents any integer of 0 or more.
* represents a bonding site in the above general formula (3). )

According to the Tag portion removal method of the present invention, the above tagged nucleoside protected body and Tag
Since a protected nucleotide is used, it is possible to selectively and rapidly remove only the Tag portion. In addition, the above effects are achieved by the carbonyl group (C=O group) directly bonded to the phenyl group in the general formula (3), etc., and the X atoms (O, S , NH, or NR ^N ) forms some kind of interaction with a reducing agent such as LiBH ₄ , making it easier for the reduction reaction to occur more favorably at the Tag compound site. Although it is speculated that it may do so, the speculation does not limit the scope of rights.

（式中、
Ｒ^１、Ｒ^２、および、Ｒ^３は、それぞれ独立して、置換されていてもよい、炭素数１０
～４０のアルキル基を表す。
Ｘは、Ｏ、Ｓ、ＮＨ、または、ＮＲ^Ｎを表す。
ｎは、１～４の整数を表す。
Ｒ^Ｎは、置換されていてもよい、炭素数１～６のアルキル基を表す。
Ｙは、下記一般式（４）、または、一般式（５）を表す。）

（式中、
Ｂａｓｅは、修飾されていてもよい核酸塩基を表す。Ｂａｓｅが２以上存在する場合に
は、複数存在するＢａｓｅは同一、または、異なっていてもよい。
Ｂは、水素原子、保護されていてもよい水酸基、または、ハロゲンを表す。Ｂが２以上
存在する場合には、複数存在するＢは、同一、または、異なっていてもよい。
Ｗは、水素原子、または、保護基を表す。
Ｒ^４は、電子求引性基を有する、炭素数１～４のアルキル基を表す。Ｒ^４が２以上存在
する場合には、複数存在するＲ^４は同一、または、異なっていてもよい。
Ｚは、Ｏ、または、Ｓを表す。Ｚが２以上存在する場合には、複数存在するＺは、同一
、または、異なっていてもよい。
ｙ１は、０以上の任意の整数を表す。
＊は、上記一般式（６）における結合箇所を表す。）

（式中、
Ｂａｓｅは、修飾されていてもよい核酸塩基を表す。Ｂａｓｅが２以上存在する場合に
は、複数存在するＢａｓｅは同一、または、異なっていてもよい。
Ｑは、水素原子、または、保護基を表す。
Ｖは、炭素数１～６のアルコキシ基、ジ（Ｃ_１－６アルキル）アミノ基、または、４位
窒素原子が保護基で保護され、さらに置換されていてもよいピペラジノ基を表す。Ｖが２
以上存在する場合には、複数存在するＶは同一、または、異なっていてもよい。
ｙ２は、０以上の任意の整数を表す。
＊は、上記一般式（６）における結合箇所を表す。） Further, the method for removing Tag moieties of the present invention preferably includes step (1) of subjecting the compound represented by general formula (6) or its derivative to reduction treatment.

(In the formula,
R ¹ , R ² and R ³ each independently have 10 carbon atoms and may be substituted.
represents an alkyl group of ∼40.
X represents O, S, NH, or ^NRN .
n represents an integer of 1-4.
R ^N represents an optionally substituted alkyl group having 1 to 6 carbon atoms.
Y represents the following general formula (4) or general formula (5). )

(In the formula,
Base represents an optionally modified nucleobase. When two or more Bases exist, the plurality of Bases may be the same or different.
B represents a hydrogen atom, an optionally protected hydroxyl group, or a halogen. When two or more B are present, the plural B may be the same or different.
W represents a hydrogen atom or a protecting group.
R ⁴ represents a C 1-4 alkyl group having an electron-withdrawing group. When two or more R ⁴ are present, the plurality of R ⁴ may be the same or different.
Z represents O or S; When there are two or more Z's, the multiple Z's may be the same or different.
y1 represents any integer of 0 or more.
* represents a bonding site in the general formula (6). )

(In the formula,
Base represents an optionally modified nucleobase. When two or more Bases exist, the plurality of Bases may be the same or different.
Q represents a hydrogen atom or a protecting group.
V represents an alkoxy group having 1 to 6 carbon atoms, a di(C _1-6 alkyl)amino group, or an optionally substituted piperazino group whose 4-position nitrogen atom is protected by a protecting group. V is 2
When there are more than one V, the plurality of V may be the same or different.
y2 represents any integer of 0 or more.
* represents a bonding site in the general formula (6). )

In the method for removing Tag moieties of the present invention, the reduction treatment preferably uses a boron-containing reducing agent or uses a boron-containing reducing agent and an amine. By having the above-described structure, selective removal of the Tag portion in the protected nucleoside tagged or protected nucleotide tagged can be easily performed.

In the method for removing Tag moieties of the present invention, the boron-containing reducing agent is preferably lithium borohydride, sodium borohydride, lithium triethylborohydride, or tetrabutylammonium borohydride. By having the above structure, the selective removal of the Tag moiety in the protected nucleoside or protected nucleotide tagged can be performed more efficiently.

The compounds or derivatives thereof (compounds (1), (2), etc., alkoxyphenyl derivatives) of the present invention are novel compounds, and by using them, the above-mentioned tagged nucleoside protected bodies and nucleotide protected bodies can be easily obtained. can be obtained.

In addition, the compounds of the present invention or derivatives thereof (compounds (3), (6), etc., protected nucleosides and protected nucleotides tagged, etc.) of the present invention have the above-mentioned structures, and are particularly suitable for liquid-phase synthesis ( Including pseudo-liquid phase synthesis), by adding a polar solvent, it can be easily separated from auxiliary materials such as amidite monomers by filtration operation, etc., and only the Tag portion can be selectively and quickly removed. It is possible to synthesize not only oligonucleotides by the liquid phase synthesis method but also protected oligonucleotides, which has been difficult in the past.

In addition, the method for producing the compounds of the present invention or derivatives thereof (compounds (7), (8), etc., oligonucleotides, etc.) is simpler than conventional methods because the above protected nucleosides and protected nucleotides are used. It enables the synthesis of oligonucleotides by a rapid liquid phase synthesis method.

Also, compounds of the present invention or derivatives thereof (compounds (1), (2), (3), (6), etc.).
Alkoxyphenyl derivatives, tagged nucleoside protected bodies, tagged nucleotide protected bodies, and the like. ) uses the tagged nucleoside protected body and the tagged nucleotide protected body, so that only the Tag moiety can be selectively and rapidly removed under mild conditions. It is possible.

BEST MODE FOR CARRYING OUT THE INVENTION Hereinafter, embodiments of the present invention will be described in detail.

（式中、
Ｒは、それぞれ独立して、置換されていてもよい、炭素数１０～４０のアルキル基を表
す。
ｍは、１～５の整数を表す。ｍが２以上の場合には、複数存在するＲＯは同一、または
、異なっていてもよい。
Ｘは、Ｏ、Ｓ、ＮＨ、または、ＮＲ^Ｎを表す。
ｎは、１～４の整数を表す。
Ｒ^Ｎは、置換されていてもよい、炭素数１～６のアルキル基を表す。） [Alkoxyphenyl compounds and derivatives thereof]
The alkoxyphenyl compounds and derivatives thereof of the present invention are compounds represented by general formula (1) or derivatives thereof.

(In the formula,
Each R independently represents an optionally substituted alkyl group having 10 to 40 carbon atoms.
m represents an integer of 1 to 5; When m is 2 or more, multiple ROs may be the same or different.
X represents O, S, NH, or ^NRN .
n represents an integer of 1-4.
R ^N represents an optionally substituted alkyl group having 1 to 6 carbon atoms. )

In the general formula (1), each R independently represents an optionally substituted alkyl group having 10 to 40 carbon atoms. The above R may be an alkyl group having 13 to 30 carbon atoms or may be an alkyl group having 15 to 20 carbon atoms, and an alkyl group having 18 carbon atoms is particularly preferable. In addition, when there are a plurality of Rs, they may each independently be a straight-chain alkyl group or a branched alkyl group, but a straight-chain alkyl group is preferable. In addition, when R is an alkyl group having 10 to 40 carbon atoms, the tagged nucleoside protected body and tagged nucleotide protected body of the present invention are easily synthesized, and the effect of improving solubility and separation ability is also obtained. be able to.

In the above general formula (1), m represents an integer of 1-5. When m is 2 or more, multiple ROs may be the same or different. m is preferably an integer of 2 to 4, more preferably m is 3. In addition, the RO group is preferably bonded to the 3-position, 4-position, or 5-position when the carbonyl group (C=O group) directly bonded to the phenyl group is the 1-position, and m =3, it is preferably attached to the 3-, 4- or 5-position. Further, by adjusting the value of m within the above range, the Ta of the present invention
It is possible to adjust the solubility of the g-nucleoside protected form and the tagged nucleotide protected form in a non-polar solvent.

In the above general formula (1), X represents O, S, NH, or ^NRN . Above all,
Preferably X is O.

In the above general formula (1), n represents an integer of 1-4. Among them, n is preferably 1 or 2.

In general formula (1) above, R 3 ^N represents an optionally substituted alkyl group having 1 to 6 carbon atoms. Examples of the R ^N include methyl group, ethyl group, 1-propyl group, 1-methyl-1-ethyl group, 1-butyl group, 1-methyl-1-propyl group, 1,1-dimethyl-1
-ethyl group, 2-methyl-1-propyl group, 1-pentyl group and 2-pentyl group.

（式中、
Ｒ^１、Ｒ^２、および、Ｒ^３は、それぞれ独立して、置換されていてもよい、炭素数１０
～４０のアルキル基を表す。
Ｘは、Ｏ、Ｓ、ＮＨ、または、ＮＲ^Ｎを表す。
ｎは、１～４の整数を表す。
Ｒ^Ｎは、置換されていてもよい、炭素数１～６のアルキル基を表す。） Further, in the present invention, the alkoxyphenyl compound and its derivative are represented by the general formula (
2) It is preferably a compound represented or a derivative thereof.

(In the formula,
R ¹ , R ² and R ³ each independently have 10 carbon atoms and may be substituted.
represents an alkyl group of ∼40.
X represents O, S, NH, or ^NRN .
n represents an integer of 1-4.
R ^N represents an optionally substituted alkyl group having 1 to 6 carbon atoms. )

In general formula (2) above, R ¹ , R ² and R ³ each independently represent an optionally substituted alkyl group having 10 to 40 carbon atoms. R ¹ , R ² and R ³ above
may be an alkyl group having 13 to 30 carbon atoms or may be an alkyl group having 15 to 20 carbon atoms, and an alkyl group having 18 carbon atoms is particularly preferable.
R ¹ , R ² and R ³ may each independently be a straight-chain alkyl group or a branched alkyl group, but all are preferably straight-chain alkyl groups.

In general formula (2) above, X, n and ^RN are the same as in general formula (1) above.

In addition, the alkoxyphenyl compounds and derivatives thereof of the present invention are exemplified in their synthesis methods in the examples, but they may be synthesized by using commercially available raw materials and using known methods.

（式中、
Ｒは、それぞれ独立して、置換されていてもよい、炭素数１０～４０のアルキル基を表
す。
ｍは、１～５の整数を表す。ｍが２以上の場合には、複数存在するＲＯは、同一、また
は、異なっていてもよい。
Ｘは、Ｏ、Ｓ、ＮＨ、または、ＮＲ^Ｎを表す。
ｎは、１～４の整数を表す。
Ｒ^Ｎは、置換されていてもよい、炭素数１～６のアルキル基を表す。
Ｙは、下記一般式（４）、または、一般式（５）を表す。）

（式中、
Ｂａｓｅは、修飾されていてもよい核酸塩基を表す。Ｂａｓｅが２以上存在する場合に
は、複数存在するＢａｓｅは同一、または、異なっていてもよい。
Ｂは、水素原子、保護されていてもよい水酸基、または、ハロゲンを表す。Ｂが２以上
存在する場合には、複数存在するＢは、同一、または、異なっていてもよい。
Ｗは、水素原子、または、保護基を表す。
Ｒ^４は、電子求引性基を有する、炭素数１～４のアルキル基を表す。Ｒ^４が２以上存在
する場合には、複数存在するＲ^４は同一、または、異なっていてもよい。
Ｚは、Ｏ、または、Ｓを表す。Ｚが２以上存在する場合には、複数存在するＺは、同一
、または、異なっていてもよい。
ｙ１は、０以上の任意の整数を表す。
＊は、上記一般式（３）における結合箇所を表す。）

（式中、
Ｂａｓｅは、修飾されていてもよい核酸塩基を表す。Ｂａｓｅが２以上存在する場合に
は、複数存在するＢａｓｅは同一、または、異なっていてもよい。
Ｑは、水素原子、または、保護基を表す。
Ｖは、炭素数１～６のアルコキシ基、ジ（Ｃ_１－６アルキル）アミノ基、または、４位
窒素原子が保護基で保護され、さらに置換されていてもよいピペラジノ基を表す。Ｖが２
以上存在する場合には、複数存在するＶは同一、または、異なっていてもよい。
ｙ２は、０以上の任意の整数を表す。
＊は、上記一般式（３）における結合箇所を表す。） [Tagged Nucleoside Protected Body, Tagged Nucleotide Protected Body, etc.]
The protected nucleoside tagged and protected nucleotide tagged in the present invention are compounds represented by general formula (3) or derivatives thereof.

(In the formula,
Each R independently represents an optionally substituted alkyl group having 10 to 40 carbon atoms.
m represents an integer of 1 to 5; When m is 2 or more, multiple ROs may be the same or different.
X represents O, S, NH, or ^NRN .
n represents an integer of 1-4.
R ^N represents an optionally substituted alkyl group having 1 to 6 carbon atoms.
Y represents the following general formula (4) or general formula (5). )

(In the formula,
Base represents an optionally modified nucleobase. When two or more Bases exist, the plurality of Bases may be the same or different.
B represents a hydrogen atom, an optionally protected hydroxyl group, or a halogen. When two or more B are present, the plural B may be the same or different.
W represents a hydrogen atom or a protecting group.
R ⁴ represents a C 1-4 alkyl group having an electron-withdrawing group. When two or more R ⁴ are present, the plurality of R ⁴ may be the same or different.
Z represents O or S; When there are two or more Z's, the multiple Z's may be the same or different.
y1 represents any integer of 0 or more.
* represents a bonding site in the above general formula (3). )

(In the formula,
Base represents an optionally modified nucleobase. When two or more Bases exist, the plurality of Bases may be the same or different.
Q represents a hydrogen atom or a protecting group.
V represents an alkoxy group having 1 to 6 carbon atoms, a di(C _1-6 alkyl)amino group, or an optionally substituted piperazino group whose 4-position nitrogen atom is protected by a protecting group. V is 2
When there are more than one V, the plurality of V may be the same or different.
y2 represents any integer of 0 or more.
* represents a bonding site in the above general formula (3). )

In general formula (3) above, R, m, X, n, and R ^N are the same as in general formula (1) above.

In the above general formula (3), Y represents the following general formula (4) or general formula (5).

In the above general formula (4), Base represents an optionally modified nucleic acid base. Ba
When two or more se exist, the plurality of Bases may be the same or different.

Examples of the Base include an adenyl group, a guanyl group, a cytidyl group, a thymidyl group, a uracil group, derivatives thereof, and modifications thereof. As the modification, for example, a polar group or the like may be protected with a known protecting group or the like. Moreover, multiple Bases present in an oligonucleotide may be the same or different.

In the above general formula (4), B is a hydrogen atom, an optionally protected hydroxyl group, or
represents halogen. When two or more B are present, the plural B may be the same or different. Moreover, in the case of a protected hydroxyl group, a known technique can be appropriately used as a protecting group and a protecting method.

In general formula (4) above, W represents a hydrogen atom or a protecting group. When W is a protecting group, known techniques can be appropriately used as protecting groups and protecting methods.

Further, W is a hydrogen atom, a trityl group, a di (C _1-6 alkoxy) trityl group, a mono (C
_1-18 alkoxy)trityl group, 9-(9-phenyl)xanthenyl group, or 9-
A (9-phenyl)thioxanthenyl group is preferred. When W is any of the above, oligonucleotides can be synthesized more easily.

In general formula (4) above, R ⁴ represents an alkyl group having 1 to 4 carbon atoms and having an electron-withdrawing group. When two or more R ⁴ are present, the plurality of R ⁴ may be the same or different. Also, ^R4 is preferably an ethyl group having an electron-withdrawing group at the 2-position. Moreover, as said electron withdrawing group, a cyano group can be mentioned as a preferable example, for example.

In the above general formula (4), Z represents O or S. When there are two or more Z's, the multiple Z's may be the same or different.

In the above general formula (4), y1 represents any integer of 0 or more. y1 is 1 to 100
, 1 to 50, 1 to 30, 1 to 20, 1 to 10, or 0.

In the above general formula (4), * represents the bonding site in the above general formula (3).

In general formula (5) above, Base is the same as in general formula (4) above.

In general formula (5) above, Q represents a hydrogen atom or a protecting group. When Q is a protecting group, known techniques can be appropriately used as protecting groups and protecting methods.

Further, Q is a hydrogen atom, a trityl group, a di (C _1-6 alkoxy) trityl group, a mono (C
_1-18 alkoxy)trityl group, 9-(9-phenyl)xanthenyl group, or 9-
A (9-phenyl)thioxanthenyl group is preferred. When Q is any of the above, oligonucleotides can be synthesized more easily.

In the above general formula (5), V is an alkoxy group having 1 to 6 carbon atoms, a di(C _1-6 alkyl)amino group, or a nitrogen atom at the 4-position protected by a protecting group, even if it is further substituted. represents a good piperazino group. When there are two or more Vs, the plurality of Vs may be the same or different.

In the above general formula (5), y2 represents any integer of 0 or more. y2 is 1 to 100
, 1 to 50, 1 to 30, 1 to 20, 1 to 10, or 0.

In the above general formula (5), * represents the bonding site in the above general formula (3).

（式中、
Ｒ^１、Ｒ^２、および、Ｒ^３は、それぞれ独立して、置換されていてもよい、炭素数１０
～４０のアルキル基を表す。
Ｘは、Ｏ、Ｓ、ＮＨ、または、ＮＲ^Ｎを表す。
ｎは、１～４の整数を表す。
Ｒ^Ｎは、置換されていてもよい、炭素数１～６のアルキル基を表す。
Ｙは、下記一般式（４）、または、一般式（５）を表す。）

（式中、
Ｂａｓｅは、修飾されていてもよい核酸塩基を表す。Ｂａｓｅが２以上存在する場合に
は、複数存在するＢａｓｅは同一、または、異なっていてもよい。
Ｂは、水素原子、保護されていてもよい水酸基、または、ハロゲンを表す。Ｂが２以上
存在する場合には、複数存在するＢは、同一、または、異なっていてもよい。
Ｗは、水素原子、または、保護基を表す。
Ｒ^４は、電子求引性基を有する、炭素数１～４のアルキル基を表す。Ｒ^４が２以上存在
する場合には、複数存在するＲ^４は同一、または、異なっていてもよい。
Ｚは、Ｏ、または、Ｓを表す。Ｚが２以上存在する場合には、複数存在するＺは、同一
、または、異なっていてもよい。
ｙ１は、０以上の任意の整数を表す。
＊は、上記一般式（６）における結合箇所を表す。）

（式中、
Ｂａｓｅは、修飾されていてもよい核酸塩基を表す。Ｂａｓｅが２以上存在する場合に
は、複数存在するＢａｓｅは同一、または、異なっていてもよい。
Ｑは、水素原子、または、保護基を表す。
Ｖは、炭素数１～６のアルコキシ基、ジ（Ｃ_１－６アルキル）アミノ基、または、４位
窒素原子が保護基で保護され、さらに置換されていてもよいピペラジノ基を表す。Ｖが２
以上存在する場合には、複数存在するＶは同一、または、異なっていてもよい。
ｙ２は、０以上の任意の整数を表す。
＊は、上記一般式（６）における結合箇所を表す。） In addition, the protected nucleoside tagged and protected nucleotide tagged according to the present invention are preferably compounds represented by general formula (6) or derivatives thereof.

(In the formula,
R ¹ , R ² and R ³ each independently have 10 carbon atoms and may be substituted.
represents an alkyl group of ∼40.
X represents O, S, NH, or ^NRN .
n represents an integer of 1-4.
R ^N represents an optionally substituted alkyl group having 1 to 6 carbon atoms.
Y represents the following general formula (4) or general formula (5). )

(In the formula,
Base represents an optionally modified nucleobase. When two or more Bases exist, the plurality of Bases may be the same or different.
B represents a hydrogen atom, an optionally protected hydroxyl group, or a halogen. When two or more B are present, the plural B may be the same or different.
W represents a hydrogen atom or a protecting group.
R ⁴ represents a C 1-4 alkyl group having an electron-withdrawing group. When two or more R ⁴ are present, the plurality of R ⁴ may be the same or different.
Z represents O or S; When there are two or more Z's, the multiple Z's may be the same or different.
y1 represents any integer of 0 or more.
* represents a bonding site in the general formula (6). )

(In the formula,
Base represents an optionally modified nucleobase. When two or more Bases exist, the plurality of Bases may be the same or different.
Q represents a hydrogen atom or a protecting group.
V represents an alkoxy group having 1 to 6 carbon atoms, a di(C _1-6 alkyl)amino group, or an optionally substituted piperazino group whose 4-position nitrogen atom is protected by a protecting group. V is 2
When there are more than one V, the plurality of V may be the same or different.
y2 represents any integer of 0 or more.
* represents a bonding site in the general formula (6). )

In general formula (6) above, R ¹ , R ² , R ³ , X, n, and ^RN are the same as in general formula (2) above.

In general formula (6) above, Y is the same as in general formula (3) above.

In addition, the tagged nucleoside protected form and the tagged nucleotide protected form of the present invention are exemplified in the examples for their synthesis methods, but they may be synthesized by other known methods using commercially available raw materials. .

Tagged nucleoside protected form and tagged nucleotide protected form production method of the present invention (
Synthetic method), for example, a method of binding the above alkoxyphenyl compound or derivative thereof to a nucleoside or nucleotide. As a binding method,
For example, a method of condensing the alkoxyphenyl compound or derivative thereof with a nucleoside or nucleotide can be used. When the condensation reaction is used, for example, the alkoxyphenyl compound represented by the general formula (1) or (2) and a nucleoside or nucleotide are combined using a condensing agent, or the reactivity is increased by a substituent or the like. A method of binding a derivative of the alkoxyphenyl compound represented by the enhanced general formula (1) or (2) (e.g., ester derivative, acid chloride derivative, etc.) with a nucleoside or nucleotide, or ) or the alkoxyphenyl compound represented by (2), and a nucleoside or nucleotide whose reactivity is enhanced by a substituent or the like, can be appropriately used for bonding.

（式中、
Ｒは、それぞれ独立して、置換されていてもよい、炭素数１０～４０のアルキル基を表
す。
ｍは、１～５の整数を表す。ｍが２以上の場合には、複数存在するＲＯは、同一、また
は、異なっていてもよい。
Ｘは、Ｏ、Ｓ、ＮＨ、または、ＮＲ^Ｎを表す。
ｎは、１～４の整数を表す。
Ｒ^Ｎは、置換されていてもよい、炭素数１～６のアルキル基を表す。
Ｙは、下記一般式（４）、または、一般式（５）を表す。）

（式中、
Ｂａｓｅは、修飾されていてもよい核酸塩基を表す。Ｂａｓｅが２以上存在する場合に
は、複数存在するＢａｓｅは同一、または、異なっていてもよい。
Ｂは、水素原子、保護されていてもよい水酸基、または、ハロゲンを表す。Ｂが２以上
存在する場合には、複数存在するＢは、同一、または、異なっていてもよい。
Ｗは、水素原子、または、保護基を表す。
Ｒ^４は、電子求引性基を有する、炭素数１～４のアルキル基を表す。Ｒ^４が２以上存在
する場合には、複数存在するＲ^４は同一、または、異なっていてもよい。
Ｚは、Ｏ、または、Ｓを表す。Ｚが２以上存在する場合には、複数存在するＺは、同一
、または、異なっていてもよい。
ｙ１は、０以上の任意の整数を表す。
＊は、上記一般式（３）における結合箇所を表す。）

（式中、
Ｂａｓｅは、修飾されていてもよい核酸塩基を表す。Ｂａｓｅが２以上存在する場合に
は、複数存在するＢａｓｅは同一、または、異なっていてもよい。
Ｑは、水素原子、または、保護基を表す。
Ｖは、炭素数１～６のアルコキシ基、ジ（Ｃ_１－６アルキル）アミノ基、または、４位
窒素原子が保護基で保護され、さらに置換されていてもよいピペラジノ基を表す。Ｖが２
以上存在する場合には、複数存在するＶは同一、または、異なっていてもよい。
ｙ２は、０以上の任意の整数を表す。
＊は、上記一般式（３）における結合箇所を表す。）

（式中、
Ｂａｓｅは、修飾されていてもよい核酸塩基を表す。Ｂａｓｅが２以上存在する場合に
は、複数存在するＢａｓｅは同一、または、異なっていてもよい。
Ｂは、水素原子、保護されていてもよい水酸基、または、ハロゲンを表す。Ｂが２以上
存在する場合には、複数存在するＢは、同一、または、異なっていてもよい。
Ｗは、水素原子、または、保護基を表す。
Ｒ^４は、電子求引性基を有する、炭素数１～４のアルキル基を表す。Ｒ^４が２以上存在
する場合には、複数存在するＲ^４は同一、または、異なっていてもよい。
Ｚは、Ｏ、または、Ｓを表す。Ｚが２以上存在する場合には、複数存在するＺは、同一
、または、異なっていてもよい。
ｙ１は、０以上の任意の整数を表す。）

（式中、
Ｂａｓｅは、修飾されていてもよい核酸塩基を表す。Ｂａｓｅが２以上存在する場合に
は、複数存在するＢａｓｅは同一、または、異なっていてもよい。
Ｑは、水素原子、または、保護基を表す。
Ｖは、炭素数１～６のアルコキシ基、ジ（Ｃ_１－６アルキル）アミノ基、または、４位
窒素原子が保護基で保護され、さらに置換されていてもよいピペラジノ基を表す。Ｖが２
以上存在する場合には、複数存在するＶは同一、または、異なっていてもよい。
ｙ２は、０以上の任意の整数を表す。） [Method for producing protected oligonucleotide, method for removing Tag portion, etc.]
The production method (synthesis method) of the protected oligonucleotide of the present invention and the Tag of the present invention
The partial removal method (substituent removal method) includes step (1) of reducing the compound represented by general formula (3) or a derivative thereof.

(In the formula,
Each R independently represents an optionally substituted alkyl group having 10 to 40 carbon atoms.
m represents an integer of 1 to 5; When m is 2 or more, multiple ROs may be the same or different.
X represents O, S, NH, or ^NRN .
n represents an integer of 1-4.
R ^N represents an optionally substituted alkyl group having 1 to 6 carbon atoms.
Y represents the following general formula (4) or general formula (5). )

(In the formula,
Base represents an optionally modified nucleobase. When two or more Bases exist, the plurality of Bases may be the same or different.
B represents a hydrogen atom, an optionally protected hydroxyl group, or a halogen. When two or more B are present, the plural B may be the same or different.
W represents a hydrogen atom or a protecting group.
R ⁴ represents a C 1-4 alkyl group having an electron-withdrawing group. When two or more R ⁴ are present, the plurality of R ⁴ may be the same or different.
Z represents O or S; When there are two or more Z's, the multiple Z's may be the same or different.
y1 represents any integer of 0 or more.
* represents a bonding site in the above general formula (3). )

(In the formula,
Base represents an optionally modified nucleobase. When two or more Bases exist, the plurality of Bases may be the same or different.
Q represents a hydrogen atom or a protecting group.
V represents an alkoxy group having 1 to 6 carbon atoms, a di(C _1-6 alkyl)amino group, or an optionally substituted piperazino group whose 4-position nitrogen atom is protected by a protecting group. V is 2
When there are more than one V, the plurality of V may be the same or different.
y2 represents any integer of 0 or more.
* represents a bonding site in the above general formula (3). )

(In the formula,
Base represents an optionally modified nucleobase. When two or more Bases exist, the plurality of Bases may be the same or different.
B represents a hydrogen atom, an optionally protected hydroxyl group, or a halogen. When two or more B are present, the plural B may be the same or different.
W represents a hydrogen atom or a protecting group.
R ⁴ represents a C 1-4 alkyl group having an electron-withdrawing group. When two or more R ⁴ are present, the plurality of R ⁴ may be the same or different.
Z represents O or S; When there are two or more Z's, the multiple Z's may be the same or different.
y1 represents any integer of 0 or more. )

(In the formula,
Base represents an optionally modified nucleobase. When two or more Bases exist, the plurality of Bases may be the same or different.
Q represents a hydrogen atom or a protecting group.
V represents an alkoxy group having 1 to 6 carbon atoms, a di(C _1-6 alkyl)amino group, or an optionally substituted piperazino group whose 4-position nitrogen atom is protected by a protecting group. V is 2
When there are more than one V, the plurality of V may be the same or different.
y2 represents any integer of 0 or more. )

In the method for producing an oligonucleotide protected body, the general formulas (3), (4), (5)
) is the same as above.

In the above general formula (7) in the method for producing the protected oligonucleotide, Ba
se, B, W, R ⁴ , Z, and y1 are the same as in general formula (4) above.

In the above general formula (8) in the method for producing an oligonucleotide protected body, Ba
se, Q, V, and y2 are the same as in general formula (5) above.

（式中、
Ｒ^１、Ｒ^２、および、Ｒ^３は、それぞれ独立して、置換されていてもよい、炭素数１０
～４０のアルキル基を表す。
Ｘは、Ｏ、Ｓ、ＮＨ、または、ＮＲ^Ｎを表す。
ｎは、１～４の整数を表す。
Ｒ^Ｎは、置換されていてもよい、炭素数１～６のアルキル基を表す。
Ｙは、下記一般式（４）、または、一般式（５）を表す。）

（式中、
Ｂａｓｅは、修飾されていてもよい核酸塩基を表す。Ｂａｓｅが２以上存在する場合に
は、複数存在するＢａｓｅは同一、または、異なっていてもよい。
Ｂは、水素原子、保護されていてもよい水酸基、または、ハロゲンを表す。Ｂが２以上
存在する場合には、複数存在するＢは、同一、または、異なっていてもよい。
Ｗは、水素原子、または、保護基を表す。
Ｒ^４は、電子求引性基を有する、炭素数１～４のアルキル基を表す。Ｒ^４が２以上存在
する場合には、複数存在するＲ^４は同一、または、異なっていてもよい。
Ｚは、Ｏ、または、Ｓを表す。Ｚが２以上存在する場合には、複数存在するＺは、同一
、または、異なっていてもよい。
ｙ１は、０以上の任意の整数を表す。
＊は、上記一般式（６）における結合箇所を表す。）

（式中、
Ｂａｓｅは、修飾されていてもよい核酸塩基を表す。Ｂａｓｅが２以上存在する場合に
は、複数存在するＢａｓｅは同一、または、異なっていてもよい。
Ｑは、水素原子、または、保護基を表す。
Ｖは、炭素数１～６のアルコキシ基、ジ（Ｃ_１－６アルキル）アミノ基、または、４位
窒素原子が保護基で保護され、さらに置換されていてもよいピペラジノ基を表す。Ｖが２
以上存在する場合には、複数存在するＶは同一、または、異なっていてもよい。
ｙ２は、０以上の任意の整数を表す。
＊は、上記一般式（６）における結合箇所を表す。）

（式中、
Ｂａｓｅは、修飾されていてもよい核酸塩基を表す。Ｂａｓｅが２以上存在する場合に
は、複数存在するＢａｓｅは同一、または、異なっていてもよい。
Ｂは、水素原子、保護されていてもよい水酸基、または、ハロゲンを表す。Ｂが２以上
存在する場合には、複数存在するＢは、同一、または、異なっていてもよい。
Ｗは、水素原子、または、保護基を表す。
Ｒ^４は、電子求引性基を有する、炭素数１～４のアルキル基を表す。Ｒ^４が２以上存在
する場合には、複数存在するＲ^４は同一、または、異なっていてもよい。
Ｚは、Ｏ、または、Ｓを表す。Ｚが２以上存在する場合には、複数存在するＺは、同一
、または、異なっていてもよい。
ｙ１は、０以上の任意の整数を表す。）

（式中、
Ｂａｓｅは、修飾されていてもよい核酸塩基を表す。Ｂａｓｅが２以上存在する場合に
は、複数存在するＢａｓｅは同一、または、異なっていてもよい。
Ｑは、水素原子、または、保護基を表す。
Ｖは、炭素数１～６のアルコキシ基、ジ（Ｃ_１－６アルキル）アミノ基、または、４位
窒素原子が保護基で保護され、さらに置換されていてもよいピペラジノ基を表す。Ｖが２
以上存在する場合には、複数存在するＶは同一、または、異なっていてもよい。
ｙ２は、０以上の任意の整数を表す。） The method for producing a protected oligonucleotide of the present invention and the method for removing a Tag moiety of the present invention preferably include step (1) of reducing the compound represented by general formula (6) or its derivative.

(In the formula,
R ¹ , R ² and R ³ each independently have 10 carbon atoms and may be substituted.
represents an alkyl group of ∼40.
X represents O, S, NH, or ^NRN .
n represents an integer of 1-4.
R ^N represents an optionally substituted alkyl group having 1 to 6 carbon atoms.
Y represents the following general formula (4) or general formula (5). )

(In the formula,
Base represents an optionally modified nucleobase. When two or more Bases exist, the plurality of Bases may be the same or different.
B represents a hydrogen atom, an optionally protected hydroxyl group, or a halogen. When two or more B are present, the plural B may be the same or different.
W represents a hydrogen atom or a protecting group.
R ⁴ represents a C 1-4 alkyl group having an electron-withdrawing group. When two or more R ⁴ are present, the plurality of R ⁴ may be the same or different.
Z represents O or S; When there are two or more Z's, the multiple Z's may be the same or different.
y1 represents any integer of 0 or more.
* represents a bonding site in the general formula (6). )

(In the formula,
Base represents an optionally modified nucleobase. When two or more Bases exist, the plurality of Bases may be the same or different.
Q represents a hydrogen atom or a protecting group.
V represents an alkoxy group having 1 to 6 carbon atoms, a di(C _1-6 alkyl)amino group, or an optionally substituted piperazino group whose 4-position nitrogen atom is protected by a protecting group. V is 2
When there are more than one V, the plurality of V may be the same or different.
y2 represents any integer of 0 or more.
* represents a bonding site in the general formula (6). )

(In the formula,
Base represents an optionally modified nucleobase. When two or more Bases exist, the plurality of Bases may be the same or different.
B represents a hydrogen atom, an optionally protected hydroxyl group, or a halogen. When two or more B are present, the plural B may be the same or different.
W represents a hydrogen atom or a protecting group.
R ⁴ represents a C 1-4 alkyl group having an electron-withdrawing group. When two or more R ⁴ are present, the plurality of R ⁴ may be the same or different.
Z represents O or S; When there are two or more Z's, the multiple Z's may be the same or different.
y1 represents any integer of 0 or more. )

(In the formula,
Base represents an optionally modified nucleobase. When two or more Bases exist, the plurality of Bases may be the same or different.
Q represents a hydrogen atom or a protecting group.
V represents an alkoxy group having 1 to 6 carbon atoms, a di(C _1-6 alkyl)amino group, or an optionally substituted piperazino group whose 4-position nitrogen atom is protected by a protecting group. V is 2
When there are more than one V, the plurality of V may be the same or different.
y2 represents any integer of 0 or more. )

In the method for producing the protected oligonucleotide and the method for removing the Tag portion of the present invention, the above general formulas (3), (4) and (5) are the same as above.

In the method for producing a protected oligonucleotide and the method for removing the Tag portion of the present invention, in the general formula (7), Base, B, W, R ⁴ , Z, and y1 are represented by the general formula (4). is similar to

In the method for producing an oligonucleotide protected body and the method for removing the Tag portion of the present invention, in the general formula (8), Base, Q, V, and y2 are represented by the general formula (5
).

In the method for producing a protected oligonucleotide and the method for removing a Tag moiety of the present invention, the reduction treatment preferably uses a boron-containing reducing agent, or uses a boron-containing reducing agent and an amine. By having the above structure, the protected oligonucleotide can be synthesized more easily.

As the boron-containing reducing agent, known ones can be used, and lithium borohydride, sodium borohydride, lithium triethylborohydride, tetrabutylammonium borohydride and the like are particularly preferable. These compounds may be used alone or in combination of two or more. By having the above structure, the protected oligonucleotide can be easily synthesized with higher yield.

When the boron-containing reducing agent and amine are used, known amines can be used as the amine, and among them, diisopropylethylamine, triethylamine, pyridine and the like are preferable. These compounds may be used alone or in combination of two or more. By having the above structure, the protected oligonucleotide can be easily synthesized with higher yield.

In the above step (1), as long as the synthesis method and reaction method do not damage the chemical structure of the compound represented by general formula (6) or the like or a derivative thereof, other known methods using commercially available raw materials can be used. good too.

In the above step (1), for example, a compound represented by general formula (6) or a derivative thereof,
A process of dissolving or mixing a boron-containing reducing agent, or a boron-containing reducing agent and an amine in an organic solvent, and stirring the mixture at -10°C or room temperature for reaction can be mentioned.

In addition, although the method for producing the protected oligonucleotide of the present invention and the method for removing the Tag portion of the present invention are illustrated in the examples, the synthesis method and the reaction method thereof are shown in the examples. method may be used.

As a method for producing a tagged oligonucleotide protected body of the present invention, for example, taking a dimer as an example, the above tagged nucleoside protected body or tagged nucleotide protected body and an amidated nucleoside or nucleotide are subjected to a coupling reaction. Let (step A),
The obtained dimer is phosphorylated in an oxidation step (step B), and the protecting group W is deprotected (
A method of obtaining a dimer of a tagged nucleotide-protected compound by performing step C) and filtration (step D) can be mentioned. Higher-order (n+1)-mers also apply the technique to n
Examples include a method of obtaining by repeating a number of times, a method of obtaining by binding nucleotide derivatives of a certain length, and the like.

Examples and the like that specifically show the configuration and effects of the present invention will be described below. Hereinafter, concentration operations in the examples were performed under reduced pressure unless otherwise specified. The evaluation items in Examples and the like were measured as follows.

<Measurement of ¹ H-NMR spectrum>
¹ H-NMR spectrum was measured using a nuclear magnetic resonance apparatus (manufactured by JEOL Ltd., product name AL400
and ECS-600 manufactured by JEOL Ltd.), using tetramethylsilane as an internal standard.

<Measurement of ¹³ C-NMR spectrum>
^{The 13} C-NMR spectrum was measured using a nuclear magnetic resonance apparatus (manufactured by JEOL Ltd., product name AL40
0 and ECS-600 manufactured by JEOL Ltd.), and tetramethylsilane as an internal standard.

<Measurement of TOF/MS spectrum>
TOF/MS spectra were measured using time-of-flight mass spectrometers (manufactured by Waters, product name LCT-PremierXE and JEOL Ltd., product name JMS T-100LP).

1. Synthesis of 2-((3,4,5-tris(octadecyloxy)benzoyl)oxy)acetic acid (A) Method via tert-butyl ester [Example 1-1]
(1) Synthesis of 2-(tert-butoxy)-2-oxoethyl 3,4,5-tris(octadecyloxy)benzoate

3,4,5-tris(octadecyloxy)benzoic acid (1.80 g, 1.94 mmol
) in THF (10 mL) was added triethylamine (0.39 g, 3.87 mmol),
After adding tert-butyl 2-bromoacetate (0.76 g, 3.87 mmol), 60
C. for 2 hours. After the reaction solution was cooled to room temperature, methanol (54 mL) was added dropwise, and the precipitated solid was filtered. The solid was washed with methanol and dried under reduced pressure at 50°C to give the title compound (1.95 g, 97%) as a white solid.
¹ H-NMR (400 MHz, CDCl ₃ ): δ0.88 (t, 9H, J=6.8 Hz)
, 1.16-1.41 (m, 84H), 1.41-1.57 (m, 6H), 1.49 (s
, 9H), 1.66-1.88 (m, 6H), 3.95-4.11 (m, 6H), 4.7
0(s, 2H), 7.31(s, 2H).
¹³ C-NMR (100 MHz, CDCl ₃ ): δ 14.02, 22.48, 22.66
, 22.83, 26.08, 26.11, 27.88, 28.08, 29.35, 29.
40, 29.56, 29.64, 29.65, 29.70, 29.97, 30.22, 3
0.36, 31.76, 31.93, 32.10, 61.66, 69.35, 73.51
, 82.32, 108.69, 108.90, 123.82, 123.93, 143.1
1, 143.38, 152.93, 165.83, 166.90.
TOF/MS (ESI): _{calcd for C67H124O7Na} [M+Na] ⁺
1063.9245, found 1063.9254.

[Example 1-2]
(2) Synthesis of 2-((3,4,5-tris(octadecyloxy)benzoyl)oxy)acetic acid

3,4,5-tris(octadecyloxy)benzoic acid 2-(tert-butoxy)-2
After adding trifluoroacetic acid (11.40 g, 100.0 mmol) to a solution of -oxoethyl (5.21 g, 5.0 mmol) in chloroform (20 mL), the mixture was stirred at 50°C for 17 hours. After cooling the reaction solution to room temperature, acetonitrile (50 mL) was added dropwise, and the precipitated solid was filtered. The solid was washed with an acetonitrile-chloroform mixed solvent and then dried under reduced pressure at 50° C. to give the title compound (4.89 g, 99%) as a white solid.
¹ H-NMR (400 MHz, CDCl ₃ ): δ0.88 (t, 9H, J=7.0 Hz)
, 1.16-1.54 (m, 90H), 1.69-1.88 (m, 6H), 3.95-4
. 11 (m, 6H), 4.87 (s, 2H), 7.30 (s, 2H).
¹³ C-NMR (100 MHz, CDCl ₃ ): δ 14.08, 22.68, 26.06
, 26.10, 29.36, 29.41, 29.57, 29.65, 29.66, 29.
72, 30.35, 31.93, 60.49, 69.30, 73.57, 108.53,
123.31, 143.17, 152.96, 165.74.
TOF/MS (ESI): _{calcd for C63H116O7Na} [M+Na] ⁺
1007.8619, found 1007.8639.

(B) Method via benzyl ester [Example 1-3]
(1) 2-benzyloxy-2-3,4,5-tris(octadecyloxy)benzoate
Synthesis of oxoethyl

3,4,5-tris(octadecyloxy)benzoic acid (0.90 g, 0.97 mmol
) in chloroform (5 mL) was added with triethylamine (0.20 g, 1.94 mmol).
) and benzyl 2-bromoacetate (0.45 g, 1.94 mmol) were added, followed by stirring at 50° C. for 14 hours. After the reaction solution was cooled to room temperature, methanol (18 mL) was added dropwise, and the precipitated solid was filtered. After washing the solid with methanol, it is dried under reduced pressure at 50°C.
The title compound (1.02 g, 97%) was obtained as a white solid.
¹ H-NMR (400 MHz, CDCl ₃ ): δ0.88 (t, 9H, J=6.8 Hz)
, 1.16-1.41 (m, 84H), 1.41-1.54 (m, 6H), 1.66-1
. 91 (m, 6H), 3.95-4.08 (m, 6H), 4.86 (s, 2H), 5.2
3 (s, 2H), 7.24-7.41 (m, 7H).
¹³ C-NMR (100 MHz, CDCl ₃ ): δ 14.08, 22.71, 26.13
, 26.16, 29.40, 29.43, 29.46, 29.62, 29.69, 29.
71, 29.76, 30.00, 30.43, 31.98, 61.24, 67.07, 6
9.40, 73.59, 108.76, 123.69, 128.32, 128.50, 1
28.65, 135.35, 143.28, 152.96, 153.01, 165.84
, 167.77.

[Example 1-4]
(2) Synthesis of 2-((3,4,5-tris(octadecyloxy)benzoyl)oxy)acetic acid

2-benzyloxy-2-oxoethyl 3,4,5-tris(octadecyloxy)benzoate (0.91 g, 0.85 mmol) in THF (4.3 mL) with 10% Pd—C
(0.09 g) was added, and the mixture was stirred at 40°C for 19 hours under a hydrogen balloon. After cooling the reaction solution to room temperature, 10% Pd—C was removed by celite filtration. After concentrating the filtrate, acetonitrile (18 mL) was added dropwise to the residue, and the precipitated solid was filtered. After washing the solid with acetonitrile, it was dried under reduced pressure at 50° C. to yield the title compound (0.84 g, 100%).
was obtained as a white solid.

2. thymidinyl-[3′→5′]deoxycytidinyl-[3′→5′]-deoxycytidinyl-[3′→5′]-deoxycytidinyl-[3′→5′]-deoxyg Anidyl
[3′→5′]-deoxycytidinyl-[3′→5′]-deoxycytidinyl-[3′→
5′]-thymidinyl-[3′→5′]-deoxyguanidyl-[3′→5′]-thymidinyl-[3′→5′]-deoxyguanidyl-[3′→5′]- deoxyadenylyl-[3
'→5']-deoxycytidinyl-[3'→5']-deoxyadenylyl-[3'→5'
]-thymidinyl-[3′→5′]-deoxyguanidyl-[3′→5′]-deoxycytidinyl-[3′→5′]-deoxyadenylyl-[3′→5′] -thymidinyl-[3'→
Synthesis of 5′]-thymidine (20mer oligonucleic acid)

[Example 2-1]
(1) Synthesis of 2-((3,4,5-tris(octadecyloxy)benzoyl)oxy)acetic acid 5′-O-(4,4′-dimethoxytrityl)thymidin-3′-yl

2-((3,4,5-tris(octadecyloxy)benzoyl)oxy)acetic acid (4.
95 g, 5.02 mmol), 5′-O-(4,4′-dimethoxytrityl)thymidine (
3.28 g, 6.03 mmol), 1-methylimidazole (2.27 g, 27.62 m
mol) in THF (50 mL) was added with COMU (4.73 g, 11.05 mmol) and stirred at room temperature for 3.5 hours. Acetonitrile (248 mL) was added dropwise to the reaction solution, and the precipitated solid was filtered. The solid was washed with an acetonitrile-THF mixed solvent and then dried under reduced pressure at 50° C. to obtain the title compound (7.16 g, 95%).
¹ H-NMR (400 MHz, CDCl ₃ ): δ0.88 (t, 9H, J=6.8 Hz)
, 1.16-1.54 (m, 93H), 1.69-1.88 (m, 6H), 2.42-2
. 58 (m, 2H), 3.42-3.55 (m, 2H), 3.78 (s, 6H), 3.9
5-4.05 (m, 6H), 4.18 (s, 1H), 4.82 (s, 2H), 5.56 (
d, 1H, J = 5.6Hz), 6.40-6.50 (m, 1H), 6.80-6.90 (
m, 4H), 7.20-7.40 (m, 9H), 7.61 (s, 1H), 8.03 (s,
1H).
¹³ C-NMR (100 MHz, CDCl ₃ ): δ 11.62, 14.10, 22.71
, 26.11, 26.15, 29.38, 29.45, 29.61, 29.68, 29.
75, 30.40, 31.96, 37.88, 55.28, 61.12, 63.68, 6
9.35, 73.60, 83.93, 84.40, 87.36, 108.59, 111.
66, 113.43, 123.26, 127.29, 128.08, 128.19, 13
0.10, 130.15, 135.20, 135.25, 135.32, 144.22,
150.11, 153.02, 158.92, 163.16, 165.85, 167.5
0.
TOF/MS ( _ESI ): _{calcd for C94H146N2O13Na} [M+
Na] ⁺ 1534.0723, found 1534.0759.

[Example 2-2]
(2) Synthesis of 2-((3,4,5-tris(octadecyloxy)benzoyl)oxy)thymidin-3′-yl acetate

2-((3,4,5-tris(octadecyloxy)benzoyl)oxy)acetic acid 5′-
O-(4,4'-dimethoxytrityl)thymidin-3'-yl (6.95 g, 4.60 m
mol) in dichloromethane solution (174 mL) was added pyrrole (1.54 g, 22.98 mm
ol), and after adding trifluoroacetic acid (0.66 g, 5.75 mmol), the mixture was stirred at room temperature for 2.5 hours.
Stirred for 5 hours. Acetone (52 mL) and acetonitrile (156 mL) were added dropwise to the reaction solution, and the precipitated solid was filtered. After washing the solid with acetonitrile-acetone-dichloromethane mixed solvent and acetonitrile, it was dried under reduced pressure at 50° C. to give the title compound (5.0
4 g, 91%).
¹ H-NMR (400 MHz, CDCl ₃ ): δ0.88 (t, 9H, J=6.8 Hz)
, 1.11-1.39 (m, 84H), 1.43-1.51 (m, 6H), 1.66 (s
, 1H), 1.71-1.85 (m, 6H), 1.95 (s, 3H), 2.36-2.5
8 (m, 3H), 3.89-3.97 (m, 2H), 3.97-4.08 (m, 6H),
4.15 (dd, 1H, J=2.0, 2.4Hz), 4.84 (s, 2H), 5.45-
5.53 (m, 1H), 6.20 (dd, 1H, J = 6.0, 8.8Hz), 7.29 (
s, 2H), 7.45 (d, 1H, J=1.2Hz), 8.59 (s, 1H).
¹³ C-NMR (100 MHz, CDCl ₃ ): δ12.58, 14.12, 22.70
, 26.07, 26.11, 29.33, 29.38, 29.43, 29.59, 29.
67, 29.68, 29.73, 30.35, 31.94, 36.99, 61.12, 6
2.57, 69.24, 73.58, 75.97, 84.91, 86.46, 108.3
6, 111.48, 123.17, 136.41, 143.09, 150.29, 152
. 96, 163.40, 165.91, 167.71.
TOF/MS (ESI): _{calcd for C73H128N2O11Na [} M+
Na] ⁺ 1231.9416, found 1231.9432.

[Example 2-3]
(3) 2-((3,4,5-tris(octadecyloxy)benzoyl)oxy)acetic acid 5′-O-((cyanoethoxy)(thymidin-3′-yl)phosphoryl)thymidine-3′
-Synthesis of yl

Under argon, 2-((3,4,5-tris(octadecyloxy)benzoyl)oxy)thymidin-3′-yl acetate (1.00 g, 0.83 mmol), 5′-O-(4,4
'-Dimethoxytrityl)thymidin-3'-yl-phosphoramidite (1.23 g, 1
. 65 mmol) in dichloromethane (15 mL) was added with 5-benzylthio-1H-tetrazole (0.32 g, 1.65 mmol), and the mixture was stirred at room temperature for 1.5 hours.
Next, cumene hydroperoxide (content 82%) (0.23 g, 1.24 mmol) was added, followed by stirring at room temperature for 2 hours. Pyrrole (0.56 g, 8.27 mmol) was added to the reaction solution.
, and trifluoroacetic acid (0.28 g, 2.48 mmol) were added, followed by stirring at room temperature for 2 hours. Acetone (15 mL) and acetonitrile (45 mL) were added dropwise to the reaction solution, and the precipitated solid was filtered. After washing the solid with acetonitrile-acetone-dichloromethane mixed solvent and acetonitrile, it was dried under reduced pressure at 50° C. to give the title compound (1.20 g, 92%
) as a white solid.
¹ H-NMR (400 MHz, CDCl ₃ ): δ0.88 (t, 9H, J=6.8 Hz)
, 1.11-1.55 (m, 90H), 1.65-2.05 (m, 12H), 2.40-
2.60 (m, 4H), 2.75-2.90 (m, 2H), 3.84 (s, 2H), 3.
95-4.10 (m, 6H), 4.15-4.50 (m, 6H), 4.85 (s, 2H)
, 5.15-5.25 (m, 1H), 5.35-5.55 (m, 1H), 6.10-6.
35 (m, 2H), 7.29 (s, 2H), 7.35 (d, 1H, J=1.2Hz), 7
. 47 (s, 1H), 9.60-9.95 (m, 2H).
¹³ C-NMR (100 MHz, CDCl ₃ ): δ12.42, 12.46, 14.10
, 19.72, 19.79, 22.71, 26.12, 26.19, 29.38, 29.
43, 29.49, 29.63, 29.70, 29.75, 30.43, 31.96, 3
6.47, 36.56, 38.46, 61.14, 61.91, 62.00, 62.63
, 62.68, 62.75, 62.80, 67.53, 69.42, 73.64, 74.
66, 74.77, 78.94, 79.12, 82.33, 82.40, 82.48, 8
5.60, 85.66, 85.79, 86.02, 86.21, 86.35, 108.6
1, 111.27, 111.33, 111.73, 111.82, 116.58, 116
. 66, 123.15, 135.87, 135.94, 136.53, 136.62, 1
43.41, 150.49, 150.59, 153.07, 163.94, 164.02
, 165.96, 167.79, 167.83.
TOF/MS (ESI): _calcd for _{C86H144N5O18PNa [ M}
+ Na] ⁺ 1589.0142, found 1589.0104.

[Example 2-4]
(4) 2-((3,4,5-tris(octadecyloxy)benzoyl)oxy)acetic acid 5′-O-((2-cyanoethoxy)(5′-O-((2-cyanoethoxy)(5 '-O
-((2-cyanoethoxy)(5'-O-((2-cyanoethoxy)(5'-O-((2
-cyanoethoxy)(5′-O-((2-cyanoethoxy)(5′-O-((2-cyanoethoxy)(5′-O-((2-cyanoethoxy)(5′-O-( (2-cyanoethoxy)(5′-O-((2-cyanoethoxy)(5′-O-((2-cyanoethoxy)(5′
-O-((2-cyanoethoxy)(5'-O-((2-cyanoethoxy)(5'-O-(
(2-cyanoethoxy)(5′-O-((2-cyanoethoxy)(5′-O-((2-cyanoethoxy)(5′-O-((2-cyanoethoxy)(5′-O -((2-cyanoethoxy)(5'-O-((2-cyanoethoxy)(thymidin-3'-yl)phosphoryl)-
N ⁴ -benzoyldeoxycytidine-3′-yl)phosphoryl)-N ⁴ -benzoyldeoxycytidine-3′-yl)phosphoryl)-N ⁴ -benzoyldeoxycytidine-3′-
yl)phosphoryl)-N ² -isobutyryldeoxyguanosin-3′-yl)phosphoryl)-N ⁴ -benzoyldeoxycytidin-3′-yl)phosphoryl)-N ⁴ -benzoyldeoxycytidin-3′-yl)phosphoryl )-thymidin-3′-yl)phosphoryl)-
N ² -isobutyryldeoxyguanosin-3′-yl)phosphoryl)thymidin-3′-yl)phosphoryl)-N ² -isobutyryldeoxyguanosin-3′-yl)phosphoryl)
-N ⁶ -benzoyldeoxyadenosine-3′-yl)phosphoryl)-N ⁴ -benzoyldeoxycytidine-3′-yl)phosphoryl)-N ⁶ -benzoyldeoxyadenosine-
3′-yl)phosphoryl)thymidin-3′-yl)phosphoryl)-N ² -isobutyryldeoxyguanosin-3′-yl)phosphoryl)-N ⁴ -benzoyldeoxycytidine-3
Synthesis of '-yl)phosphoryl)-N ⁶ -benzoyldeoxyadenosin-3'-yl)phosphoryl)thymidin-3'-yl)phosphoryl)thymidin-3'-yl

The same procedure as in Example 2-3 was repeated 18 times to give the title compound (1.10 g).
TOF/MS (ESI): m/z 3114.3 [M-3H] ^3- , 2335.6 [M-4
H] ^4- , 1868.9 [M-5H] ^5- , 1557.6 [M-6H] ^6- .

[Example 2-5]
(5) Deprotection and Purification Step The compound (50 mg, 5.4 μmol) synthesized in Example 2-4 was added with 28% aqueous ammonia (
2.1 mL) was added, and the mixture was heated and stirred at 80° C. for 1.5 hours. After the reaction solution was concentrated under reduced pressure, 0.1 mol/L-triethylamine acetate buffer was added to the concentrated residue, and the precipitated insoluble matter was removed by filtration. The eluate obtained by C-18 cartridge purification of the filtrate is dried to give the target thymidinyl-[3′→5′]deoxycytidinyl-[3′→
5′]-deoxycytidinyl-[3′→5′]-deoxycytidinyl-[3′→5′]-
Deoxyguanidyl-[3′→5′]-deoxycytidinyl-[3′→5′]-deoxycytidinyl-[3′→5′]-thymidinyl-[3′→5′]-deoxy guanidyl-[3
'→5']-thymidinyl-[3'→5']-deoxyguanidyl-[3'→5']-deoxyadenylyl-[3'→5']-deoxycytidinyl-[3' →5′]-deoxyadenylyl-[3′→5′]-thymidinyl-[3′→5′]-deoxyguanidyl-[3′→
5′]-deoxycytidinyl-[3′→5′]-deoxyadenylyl-[3′→5′]-
Thymidinyl-[3′→5′]-thymidine was obtained.
HPLC (shodex ODP (4.6 Φ × 150 mm), flow rate: 1.0 mL / min,
Transfer: 0.1 mol/L triethylamine-acetate buffer, acetonitrile gradie
nt: 0-15 min (2 to 98% acetonitrile), Rt = 3.9 min (98.6
%)
TOF/MS (ESI): m/z 3020.9 [M-2H] ^2- , 2013.4 [M-3
H] ^3- , 1509.8 [M-4H] ^4- .

(6) Synthesis of 2-((3,4,5-tris(octadecyloxy)benzoyl)oxy)acetic acid 5′-O-(4,4′-dimethoxytrityl)thymidin-3′-yl [Example 2- 6]
(When using EDCI-HCl as a condensing agent)

2-((3,4,5-tris(octadecyloxy)benzoyl)oxy)acetic acid (0.
87 g, 0.89 mmol), 5′-O-(4,4′-dimethoxytrityl)thymidine (
4-dimethylaminopyridine (0.02 g, 0.18 mmol) and 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride (0.34 g , 1.77 mmol) was added, and the mixture was stirred at room temperature for 14 hours. Acetonitrile (18 mL) was added dropwise to the reaction solution, and the precipitated solid was filtered. The solid was washed with an acetonitrile-THF mixed solvent and then dried under reduced pressure at 50° C. to obtain a crude product (1.28 g). The title compound (1.06 g, 79%
) as a white solid.

[Example 2-7]
(When using CDI as a condensing agent)

2-((3,4,5-tris(octadecyloxy)benzoyl)oxy)acetic acid (9.
86 g, 10.0 mmol), 4-dimethylaminopyridine (0.12 g, 1.0 mmol
l) 1,1′-carbonyldiimidazole (2
. 43 g, 15.0 mmol) was added, and the mixture was stirred at room temperature for 1 hour and then stirred at 30 to 35° C. for 4 hours. Then 5′-O-(4,4′-dimethoxytrityl)thymidine (10.89
g, 20.0 mmol) was added and stirred at room temperature for 17 hours. Furthermore, 5'-O-
After adding (4,4′-dimethoxytrityl)thymidine (1.09 g, 0.4 mmol), the mixture was stirred at room temperature for 4.5 hours. After adding dichloromethane (10 mL) to the reaction solution, acetonitrile (400 mL) was added dropwise, and the precipitated solid was filtered. After washing the solid with an acetonitrile-dichloromethane mixed solvent, it was dried under reduced pressure at 50° C. to give a crude product (14.4
4g) was obtained. The title compound (11.11 g, 74%
) as a white solid.

3.2-((3,4,5-tris(octadecyloxy)benzoyl)oxy)acetic acid 5
'-O-((2-cyanoethoxy)(5'-O-((2-cyanoethoxy)(5'-O-
Synthesis of (4,4′-dimethoxytrityl)-N ² -isobutyryldeoxyguanosin-3′-yl)phosphoryl)thymidin-3′-yl)phosphoryl)-N ⁶ -benzoyldeoxyadenosin-3′-yl

[Example 3-1]
(1) 2-((3,4,5-tris(octadecyloxy)benzoyl)oxy)acetic acid 5′-O-(4,4′-dimethoxytrityl)-N ⁶ -benzoyldeoxyadenosine-
Synthesis of 3'-yl

2-((3,4,5-Tris(octadecyloxy)benzoyl)oxy)acetic acid (1.97 g, 2.00 mmol) and 5′-O-(4,4′- dimethoxytrityl)-N ⁶ -benzoyldeoxyadenosine (1.58 g, 2.40 mm
ol) gave the title compound (3.11 g, 96%).
¹ H-NMR (400 MHz, CDCl ₃ ): δ0.88 (t, 9H, J=6.8 Hz)
, 1.11-1.40 (m, 84H), 1.42-1.50 (m, 6H), 1.71-1
. 84 (m, 7H), 2.73 (dd, 1H, J = 4.4, 12.8Hz), 3.04-
3.13 (m, 1H), 3.46 (d, 2H, J=4.0Hz), 3.77 (s, 6H)
, 3.99-4.05 (m, 6H), 4.34-4.37 (m, 1H), 4.86 (s,
2H), 5.67 (d, 1H, J=5.6Hz), 6.51 (dd, 1H, J=5.6,
8.4Hz), 6.79 (d, 2H, J = 9.4Hz), 7.18-7.29 (m, 7H
), 7.31 (s, 2H), 7.38 (d, 2H, J = 7.2 Hz), 7.53 (dd,
2H, J = 7.2, 7.6Hz), 7.59-7.63 (m, 1H), 8.03 (d, 2
H, J = 7.2 Hz), 8.17 (s, 1H), 8.74 (s, 1H), 8.98 (s,
1H).
¹³ C-NMR (100 MHz, CDCl ₃ ): δ 14.13, 22.71, 26.08
, 26.11, 29.33, 29.38, 29.43, 29.60, 29.67, 29.
74, 30.36, 31.94, 38.00, 55.23, 61.16, 63.52, 6
9.23, 73.58, 84.35, 84.48, 86.82, 108.35, 113.
25, 123.22, 123.32, 127.03, 127.85, 127.95, 12
8.08, 128.91, 129.99, 130.04, 132.82, 133.66,
135.38, 135.42, 141.21, 143.07, 144.33, 149.5
3, 151.56, 152.75, 152.97, 158.63, 164.48, 165
. 89, 167.38.
TOF/MS ( _ESI ): calcd for _{C101H149N5O12Na [ M}
+ Na] ⁺ 1647.1100, found 1647.1149.

[Example 3-2]
(2) Synthesis of 2-((3,4,5-tris(octadecyloxy)benzoyl)oxy)acetic acid N ⁶ -benzoyldeoxyadenosin-3′-yl

2-((3,4,5-tris(octadecyloxy)benzoyl)oxy)acetic acid 5′-O-(4,4′-dimethoxytrityl)-N ⁶ -benzoyldeoxyadenosine according to the method of Example 2-2 -3′-yl (3.00 g, 1.85 mmol) gave the title compound (2.25 g, 92%).
¹ H-NMR (400 MHz, CDCl ₃ ): δ0.88 (t, 9H, J=6.8 Hz)
, 1.11-1.40 (m, 84H), 1.44-1.50 (m, 6H), 1.71-1
. 85 (m, 7H), 2.53 (dd, 1H, J = 5.6, 14.0Hz), 3.21-
3.28 (m, 1H), 3.89-3.96 (m, 1H), 3.98-4.06 (m, 6
H), 4.36 (s, 1H), 4.87 (s, 2H), 5.70 (d, 1H, J = 5.2
Hz), 5.91 (brs, 1H), 6.32 (dd, 1H, J = 5.6, 10.0Hz
), 7.32 (s, 2H), 7.54 (dd, 2H, J = 7.2, 8.0 Hz), 7.6
1-7.65 (m, 1H), 8.03 (d, 2H, J=7.2Hz), 8.07 (s, 1
H), 8.79 (s, 1H), 9.07 (s, 1H).
¹³ C-NMR (100 MHz, CDCl ₃ ): δ 14.12, 22.70, 26.07
, 26.11, 29.33, 29.37, 29.42, 29.59, 29.67, 29.
73, 30.35, 31.94, 37.76, 61.20, 63.17, 69.28, 7
3.60, 87.18, 87.61, 108.39, 123.17, 124.66, 12
7.90, 128.96, 133.01, 133.39, 142.42, 143.17,
150.40, 150.68, 152.26, 153.00, 164.42, 165.9
7, 167.41.
TOF/MS ( _ESI ): _calcd for _{C80H131N5O10Na [} M+
Na] ⁺ 1344.9794, found 1344.9828.

[Example 3-3]
(3) 5′-O-((2-cyanoethoxy)(thymidin-3′-yl)phosphoryl)-N ⁶ -benzoyl 2-((3,4,5-tris(octadecyloxy)benzoyl)oxy)acetic acid Synthesis of deoxyadenosin-3′-yl

2-((3,4,5-Tris(octadecyloxy)benzoyl)oxy)acetic acid N ⁶ -benzoyldeoxyadenosin-3′-yl (2.00 g) was prepared according to the method of Example 2-3.
, 1.51 mmol) and DMTr-dT-CE-phosphoramidite (2.25 g, 3.
02 mmol) gave the title compound (2.50 g, 97%).
¹ H-NMR (400 MHz, CDCl ₃ ): δ0.88 (t, 9H, J=6.8 Hz)
, 1.12-1.40 (m, 84H), 1.43-1.50 (m, 6H), 1.71-1
. 85 (m, 9H), 1.94 (s, 1H), 2.22-2.51 (m, 2H), 2.7
0-2.78 (m, 3H), 3.14-3.26 (m, 1H), 3.45-3.56 (m
, 1H), 3.69-3.85 (m, 2H), 3.95-4.07 (m, 6H), 4.0
7-4.32 (m, 3H), 4.40-4.48 (m, 3H), 4.88 (s, 2H),
4.99-5.11 (m, 1H), 5.67-5.73 (m, 1H), 6.11-6.2
2 (m, 1H), 6.50-6.58 (m, 1H), 7.30 (s, 2H), 7.41-
7.53 (m, 3H), 7.57-7.61 (m, 1H), 8.02-8.06 (m, 2
H), 8.38 (m, 1H), 8.81 (m, 1H), 9.58-9.67 (m, 2H)
.
¹³ C-NMR (100 MHz, CDCl ₃ ): δ12.43, 14.14, 19.66
, 19.73, 19.78, 22.71, 26.09, 26.15, 29.35, 29.
38, 29.46, 29.61, 29.68, 29.74, 29.99, 30.38, 3
1.94, 36.57, 36.87, 38.35, 61.13, 61.84, 61.92
, 62.45, 62.49, 62.58, 62.62, 67.39, 69.27, 73.
61, 74.81, 75.06, 78.89, 79.19, 79.24, 82.98, 8
3.05, 83.12, 84.47, 84.54, 85.43, 85.48, 85.62
, 85.67, 85.79, 85.93, 108.34, 111.22, 111.27,
116.52, 116.64, 123.01, 123.82, 124.08, 128.2
7, 128.31, 128.69, 128.72, 132.87, 132.94, 133
. 17, 133.41, 136.45, 141.68, 141.86, 143.18, 1
49.93, 150.00, 150.47, 151.84, 152.05, 152.67
, 153.01, 163.81, 163.86, 165.43, 165.53, 165.
99, 167.76.
TOF/MS ₍ ESI): _calcd for _{C93H147N8O17PNa [} M
+ Na] ⁺ 1702.0520, found 1702.0604.

[Example 3-4]
(4) 2-((3,4,5-tris(octadecyloxy)benzoyl)oxy)acetic acid 5′-O-((2-cyanoethoxy)(5′-O-((2-cyanoethoxy)(5 '-O
-(4,4'-dimethoxytrityl)-N ² -isobutyryldeoxyguanosine-3'-
Synthesis of yl)phosphoryl)thymidin-3′-yl)phosphoryl)-N ⁶ -benzoyldeoxyadenosin-3′-yl

2-((3,4,5-tris(octadecyloxy)benzoyl)oxy)acetic acid 5′-
O-((2-cyanoethoxy)(thymidin-3′-yl)phosphoryl)-N ⁶ -benzoyldeoxyadenosin-3′-yl (7.98 g, 4.75 mmol), N ² -isobutyryl-DMTr-dG- CE phosphoramidite (11.97 g, 14.25 mmol)
5-benzylthio-1H-tetrazole (2.7
4 g, 14.25 mmol) was added, and the mixture was stirred at room temperature for 3 hours. Next, cumene hydroperoxide (content 82%) (2.64 g, 14.25 mmol) was added, followed by 1
Stirred for an hour. Acetone (120 mL) and acetonitrile (359 mL) were added dropwise to the reaction solution, and the precipitated solid was filtered. The solid was washed with a mixed solvent of acetonitrile-acetone-dichloromethane, acetonitrile and methanol, and dried under reduced pressure at 50° C. to obtain the title compound (10.86 g, 94%).
¹ H-NMR (400 MHz, CDCl ₃ ): δ0.88 (t, 9H, J=6.8 Hz)
, 1.12-1.40 (m, 90H), 1.43-1.50 (m, 6H), 1.71-1
. 93 (m, 9H), 1.95-2.16 (m, 2H), 2.35-2.87 (m, 9H
), 3.16-3.37 (m, 3H), 3.76 (s, 6H), 3.99-4.05 (m
, 6H), 4.09-4.43 (m, 10H), 4.86 (s, 2H), 5.07-5.
26 (m, 2H), 5.65-5.71 (m, 1H), 5.91-6.22 (m, 2H)
, 6.49-6.54 (m, 1H), 6.78 (d, 4H, J=8.4Hz), 7.15
-7.27 (m, 8H), 7.30 (s, 2H), 7.34-7.36 (m, 2H), 7
. 43-7.48 (m, 2H), 7.54-7.59 (m, 1H), 7.69-7.71
(m, 1H), 7.98-8.02 (m, 2H), 8.30-8.41 (m, 1H), 8
. 75-8.82 (m, 1H), 9.43-9.58 (m, 1H), 9.88-10.2
7 (m, 1H), 12.09-12.22 (m, 1H).
¹³ C-NMR (100 MHz, CDCl ₃ ): δ12.23, 12.36, 14.13
, 18.84, 19.00, 19.05, 19.65, 19.73, 22.70, 26.
09, 26.15, 29.35, 29.37, 29.46, 29.61, 29.68, 2
9.74, 29.77, 30.38, 31.94, 35.89, 36.47, 36.63
, 55.26, 61.13, 62.47, 62.63, 62.68, 63.42, 67.
55, 69.28, 73.61, 74.86, 75.05, 79.76, 81.43, 8
3.08, 83.59, 84.68, 86.85, 108.33, 111.11, 111
. 17, 113.15, 113.26, 116.48, 116.75, 116.88, 1
21.42, 121.59, 123.03, 124.05, 127.07, 127.12
, 127.78, 127.85, 127.98, 128.06, 128.74, 129.
14, 130.05, 132.88, 133.37, 135.23, 139.48, 14
1.92, 142.01, 143.18, 144.23, 147.35, 148.11,
148.27, 148.49, 149.98, 150.09, 151.80, 152.6
0, 153.01, 155.40, 155.47, 158.62, 158.68, 164
. 77, 165.20, 165.96, 167.71, 167.74, 179.81, 1
80.21.
TOF/MS _{( ESI} ): _{calcd for C131H186N14O26P2Na}
[M+Na] ⁺ 2456.3036, found 2456.2988.

2-((3,4,5-tris(octadecyloxy)benzoyl)oxy)acetic acid 5′-
O-((2-cyanoethoxy)(5'-O-((2-cyanoethoxy)(5'-O-(4
,4′-dimethoxytrityl)thymidin-3′-yl)phosphoryl)-N ² -isobutyryldeoxyguanosin-3′-yl)phosphoryl)-N ⁴ -benzoyldeoxycytidin-3′-yl
[Example 3-5]
(1) 2-((3,4,5-tris(octadecyloxy)benzoyl)oxy)acetic acid 5′-O-(4,4′-dimethoxytrityl)-N ⁴ -benzoyldeoxycytidine-3
'-yl synthesis

2-((3,4,5-Tris(octadecyloxy)benzoyl)oxy)acetic acid (1.97 g, 2.00 mmol) and 5′-O-(4,4′- dimethoxytrityl)-N ⁴ -benzoyldeoxycytidine (1.52 g, 2.40 mmo
l) gave the title compound (3.10 g, 97%).
¹ H-NMR (400 MHz, CDCl ₃ ): δ0.88 (t, 9H, J=6.8 Hz)
, 1.12-1.53 (m, 90H), 1.71-1.85 (m, 6H), 2.36-2
. 43 (m, 1H), 2.83-2.95 (m, 1H), 3.49 (ddd, 2H, J =
3.2, 10.4, 18.0 Hz), 3.78 (s, 3H), 3.79 (s, 3H), 3
. 95-4.07 (m, 6H), 4.30-4.35 (m, 1H), 4.83 (dd, 2
H, J = 16.0, 23.6Hz), 5.51-5.58 (m, 1H), 6.33 (dd
, 1H, J = 6.0, 7.6Hz), 6.86 (dd, 4H, J = 2.4, 8.8Hz)
, 7.20-7.38 (m, 11H), 7.50-7.54 (m, 2H), 7.60-7
. 64 (m, 1H), 7.88 (d, 2H, J=7.2Hz), 8.14 (d, 1H, J
= 7.6Hz), 8.40-8.60 (brs, 1H).
¹³ C-NMR (100 MHz, CDCl ₃ ): δ 14.09, 22.70, 26.10
, 26.15, 29.38, 29.46, 29.61, 29.68, 29.75, 30.
40, 31.95, 39.47, 55.26, 61.10, 63.17, 69.33, 7
3.59, 75.77, 84.68, 87.32, 108.54, 113.46, 123
. 32, 127.22, 128.10, 128.13, 129.10, 130.04, 1
30.10, 133.20, 135.13, 135.30, 144.07, 153.01
, 158.84, 165.81, 167.43.
TOF/MS ( _ESI ): calcd for _{C100H149N3O13Na [ M}
+ Na] ⁺ 1623.0988, found 1623.1069.

[Example 3-6]
(2) Synthesis of N ⁴ -benzoyldeoxycytidin-3′-yl 2-((3,4,5-tris(octadecyloxy)benzoyl)oxy)acetate

2-((3,4,5-tris(octadecyloxy)benzoyl)oxy)acetic acid 5′-O-(4,4′-dimethoxytrityl)-N ⁶ -benzoyldeoxycytidine according to the method of Example 2-2 -3′-yl (3.04 g, 1.90 mmol) to the title compound (
2.39 g, 97%).
¹ H-NMR (400 MHz, CDCl ₃ ): δ0.88 (t, 9H, J=6.8 Hz)
, 1.20-1.51 (m, 90H), 1.71-1.85 (m, 6H), 2.46-2
. 53 (m, 1H), 2.69-2.80 (m, 1H), 3.92-4.04 (m, 8H
), 4.27 (d, 1H, J = 2.4 Hz), 4.84 (dd, 2H, J = 16.4, 2
0.4 Hz), 5.51-5.53 (m, 1H), 6.26 (dd, 1H, J = 5.6,
7.6 Hz), 7.29 (s, 2H), 7.48-7.62 (m, 4H), 7.88 (d
, 2H, J = 7.6 Hz), 8.28 (d, 1H, J = 7.2 Hz), 8.66-8.9
1 (brs, 1H).
¹³ C-NMR (100 MHz, CDCl ₃ ): δ14.10, 22.71, 26.10
, 26.14, 29.38, 29.45, 29.61, 29.68, 29.75, 30.
39, 31.95, 38.46, 61.15, 62.37, 69.34, 73.61, 7
5.80, 85.82, 88.44, 108.55, 123.25, 127.61, 12
9.07, 133.25, 143.25, 145.32, 153.01, 162.38,
165.89, 167.79.
TOF/MS ( _ESI ): _{calcd for C79H131N3O11Na} [M+
Na] ⁺ 1320.9681, found 1320.9723.

[Example 3-7]
(3) 2-((3,4,5-tris(octadecyloxy)benzoyl)oxy)acetic acid 5′-O-((2-cyanoethoxy)(N ² -isobutyryldeoxyguanosine-3′-
Synthesis of yl)phosphoryl)-N ⁴ -benzoyldeoxycytidin-3′-yl

2-((3,4,5-Tris(octadecyloxy)benzoyl)oxy)acetic acid N ⁴ -benzoyldeoxycytidin-3′-yl (2.27 g,
1.75 mmol) and N ² -isobutyrylDMTr-dG-CE-phosphoramidite (
2.94 g, 3.50 mmol) gave the title compound (3.02 g, 99%).
¹ H-NMR (400 MHz, CDCl ₃ ): δ0.88 (t, 9H, J=6.8 Hz)
, 1.08-1.53 (m, 96H), 1.70-1.84 (m, 6H), 2.36-2
. 62 (m, 2H), 2.75-3.30 (m, 7H), 3.75-3.87 (m, 2H
), 3.98-4.04 (m, 6H), 4.26-4.53 (m, 6H), 4.78-4
. 89 (m, 2H), 5.26-5.29 (m, 1H), 5.42-5.52 (m, 1H
), 5.92-6.11 (m, 1H), 6.24-6.38 (m, 1H), 7.26-7
. 28 (m, 2H), 7.38-7.67 (m, 4H), 7.80-7.89 (m, 2H
), 7.95-8.00 (m, 1H), 8.14-8.24 (m, 1H), 10.22-
10.70 (m, 1H), 12.08-12.22 (m, 1H).
¹³ C-NMR (100 MHz, CDCl ₃ ): δ 14.14, 18.96, 18.99
, 19.05, 19.81, 19.87, 22.71, 26.08, 26.15, 29.
34, 29.38, 29.46, 29.61, 29.68, 29.75, 30.37, 3
1.94, 36.01, 36.06, 38.37, 38.83, 39.11, 61.09
, 62.01, 62.59, 62.64, 62.77, 62.82, 67.20, 67.
51, 69.25, 73.60, 74.76, 75.03, 80.49, 83.50, 8
3.75, 84.57, 84.81, 86.32, 87.91, 88.04, 97.29
, 97.50, 108.30, 116.55, 116.68, 120.72, 121.0
5, 123.01, 127.73, 127.80, 128.88, 128.93, 132
. 71, 132.81, 133.28, 138.19, 138.30, 143.13, 1
44.20, 144.42, 147.79, 147.99, 148.16, 148.33
, 152.99, 155.25, 155.65, 162.86, 163.04, 165.
92, 165.95, 167.77, 167.85, 179.77, 180.03.
TOF/MS (ESI): _calcd for _{C96H152N9O18PNa [ M}
+ Na] ⁺ 1773.0891, found 1773.0914.

[Example 3-8]
(4) 2-((3,4,5-tris(octadecyloxy)benzoyl)oxy)acetic acid 5′-O-((2-cyanoethoxy)(5′-O-((2-cyanoethoxy)(5 '-O
Synthesis of -(4,4'-dimethoxytrityl)thymidin-3'-yl)phosphoryl)-N ² -isobutyryldeoxyguanosin-3'-yl)phosphoryl)-N ⁴ -benzoyldeoxycytidin-3'-yl

2-((3,4,5-Tris(octadecyloxy)benzoyl)oxy)acetic acid 5′-O-((2-cyanoethoxy)(N ² -isobutyryldeoxyguanosine) according to the method of Example 3-4 -3′-yl)phosphoryl)-N ⁴ -benzoyldeoxycytidine-3′
-yl (2.80 g, 1.60 mmol) and DMTr-dT-CE phosphoramidite (
3.58 g, 4.80 mmol) gave the title compound (3.44 g, 89%).
¹ H-NMR (400 MHz, CDCl ₃ ): δ0.88 (t, 9H, J=6.8 Hz)
, 1.13-1.55 (m, 99H), 1.71-1.84 (m, 6H), 1.95-2
. 14 (m, 2H), 2.27-2.95 (m, 9H), 3.00-3.55 (m, 3H
), 3.73-3.81 (m, 6H), 3.99-4.04 (m, 6H), 4.14-4
. 48 (m, 11H), 4.82-4.85 (m, 2H), 5.05-5.53 (m, 3
H), 6.00-6.46 (m, 3H), 6.76-6.87 (m, 4H), 7.20-
7.36 (m, 11H), 7.44-7.64 (m, 4H), 7.72-7.88 (m,
3H), 8.11-8.22 (m, 1H), 9.00-9.15 (m, 1H), 10.2
9-10.49 (m, 1H), 12.14-12.17 (m, 1H).
¹³ C-NMR (100 MHz, CDCl ₃ ): δ 11.67, 11.71, 14.13
, 18.86, 18.90, 18.94, 18.98, 19.00, 19.59, 19.
67, 19.81, 19.88, 22.70, 26.08, 26.15, 29.35, 2
9.37, 29.46, 29.61, 29.67, 29.74, 29.77, 30.37
, 31.94, 35.88, 38.67, 55.29, 61.13, 62.66, 62.
73, 62.87, 63.33, 69.25, 73.59, 84.20, 84.28, 8
5.31, 87.29, 108.31, 111.69, 111.73, 113.15, 1
13.35, 116.64, 116.79, 123.01, 127.31, 127.71
, 127.77, 128.08, 128.99, 129.14, 130.09, 133.
33, 134.93, 134.97, 135.01, 135.17, 143.14, 14
3.96, 144.04, 147.84, 147.93, 148.16, 150.40,
150.48, 152.99, 155.48, 158.83, 163.52, 165.9
6, 167.86, 179.65, 179.76.
TOF/MS _{( ESI} ): _{calcd for C130H186N12O27P2Na}
[M+Na] ⁺ 2432.2923, found 2432.2937.

2-((3,4,5-tris(octadecyloxy)benzoyl)oxy)acetic acid 5′-
O-((2-cyanoethoxy)(5'-O-((2-cyanoethoxy)(5'-O-(4
,4′-dimethoxytrityl)thymidin-3′-yl)phosphoryl)thymidin-3′-yl)phosphoryl)-N ² -isobutyryldeoxyguanosin-3′-yl
[Example 3-9]
(1) 2-((3,4,5-tris(octadecyloxy)benzoyl)oxy)acetic acid 5′-O-(4,4′-dimethoxytrityl)-N ² -isobutyryldeoxyguanosine-3′- Synthesis of yl

2-((3,4,5-Tris(octadecyloxy)benzoyl)oxy)acetic acid (1.97 g, 2.00 mmol) and 5′-O-(4,4′- dimethoxytrityl)-N ² -isobutyryldeoxyguanosine (1.54 g, 2.40 m
mol) gave the title compound (3.14 g, 98%).
¹ H-NMR (400 MHz, CDCl ₃ ): δ 0.86-0.89 (m, 12H), 1
. 03 (d, 3H, J = 6.8Hz), 1.11-1.40 (m, 84H), 1.42-
1.50 (m, 6H), 1.70-1.83 (m, 6H), 1.95 (sept, 1H,
J = 6.8 Hz), 2.51 (dd, 1H, J = 5.2, 12.4 Hz), 3.17-3
. 29 (m, 2H), 3.42 (dd, 1H, J = 3.2, 10.4Hz), 3.76 (
s, 3H), 3.77 (s, 3H), 3.98-4.05 (m, 6H), 4.22 (s,
1H), 4.83 (s, 2H), 5.70 (d, 1H, J = 6.0 Hz), 6.07 (d
d, 1H, J = 5.2, 9.2Hz), 6.76-6.82 (m, 4H), 7.16-7
. 38 (m, 9H), 7.43-7.50 (m, 2H), 7.71 (s, 1H), 7.7
3(s, 1H), 11.96(s, 1H).
¹³ C-NMR (100 MHz, CDCl ₃ ): δ 14.13, 18.64, 18.77
, 22.71, 26.05, 26.11, 29.32, 29.37, 29.42, 29.
60, 29.66, 29.68, 29.74, 29.75, 29.77, 30.33, 3
1.94, 36.17, 37.21, 55.24, 61.24, 63.48, 69.30
, 73.64, 76.16, 83.94, 84.44, 86.49, 108.33, 11
3.27, 113.29, 122.34, 123.24, 127.15, 128.02,
129.99, 135.47, 135.77, 137.83, 143.04, 144.7
0, 147.18, 148.02, 152.94, 155.38, 158.74, 166
. 00, 167.35, 178.30.
TOF/MS ( _ESI ): _{calcd for C98H151N5O13Na} [M+
Na] ⁺ 1629.1206, found 1629.1240.

[Example 3-10]
(2) Synthesis of 2-((3,4,5-tris(octadecyloxy)benzoyl)oxy)acetic acid N ² -isobutyryldeoxyguanosin-3′-yl

2-((3,4,5-tris(octadecyloxy)benzoyl)oxy)acetic acid 5′-O-(4,4′-dimethoxytrityl)-N ² -isobutyryl according to the method of Example 2-2 Deoxyguanosin-3′-yl (3.05 g, 1.90 mmol) gave the title compound (2.42 g, 98%).
¹ H-NMR (400 MHz, CDCl ₃ ): δ0.88 (t, 9H, J=6.8 Hz)
, 1.20-1.50 (m, 96H), 1.71-1.85 (m, 6H), 2.48 (d
d, 1H, J = 5.6, 14.0Hz), 2.72 (sept, 1H, J = 6.8Hz)
, 2.97-3.05 (m, 1H), 3.49 (brs, 2H), 3.86-4.05 (
m, 8H), 4.26 (s, 1H), 4.86 (s, 2H), 5.59 (d, 1H, J =
6.0), 6.13 (dd, 1H, J = 5.6, 9.2Hz), 7.30 (s, 2H),
7.86 (s, 2H), 8.82 (s, 1H), 12.16 (s, 1H).
¹³ C-NMR (100 MHz, CDCl ₃ ): δ 14.10, 18.94, 18.97
, 22.71, 26.09, 26.15, 29.38, 29.45, 29.61, 29.
69, 29.75, 30.39, 31.96, 36.50, 37.85, 61.26, 6
2.92, 69.43, 73.67, 86.27, 86.34, 108.59, 122.
49, 123.26, 138.42, 143.34, 147.09, 147.73, 15
3.04, 155.01, 166.02, 167.52, 178.69.
TOF/MS (ESI): _{calcd for C77H133N5O11Na [} M+
Na] ⁺ 1326.9899, found 1326.9879.

[Example 3-11]
(3) 2-((3,4,5-tris(octadecyloxy)benzoyl)oxy)acetic acid 5′-O-((2-cyanoethoxy)(thymidin-3′-yl)phosphoryl)-N ² -iso Synthesis of butyryldeoxyguanosin-3′-yl

2-((3,4,5-Tris(octadecyloxy)benzoyl)oxy)acetic acid N ² -isobutyryldeoxyguanosin-3′-yl (2.25
g, 1.72 mmol) and DMTr-dT-CE-phosphoramidite (2.57 g, 3
. 45 mmol) gave the title compound (2.68 g, 94%).
¹ H-NMR (400 MHz, CDCl ₃ ): δ0.88 (t, 9H, J=6.8 Hz)
, 1.05-1.52 (m, 96H), 1.71-1.89 (m, 9H), 2.24-2
. 78 (m, 8H), 3.38-3.49 (m, 1H), 3.69-4.05 (m, 8H
), 4.17-4.89 (m, 7H), 5.20-5.29 (m, 1H), 5.55-5
. 66 (m, 1H), 6.08-6.24 (m, 2H), 7.30 (s, 2H), 7.4
2-7.54 (m, 1H), 7.68-7.73 (m, 1H), 8.64-8.83 (m
, 1H), 10.20-10.32 (m, 1H), 12.11-12.22 (m, 1H)
.
¹³ C-NMR (100 MHz, CDCl ₃ ): δ12.46, 14.14, 18.59
, 18.93, 19.06, 19.42, 19.66, 19.71, 19.73, 19.
78, 22.71, 26.07, 26.14, 29.33, 29.38, 29.45, 2
9.61, 29.68, 29.74, 30.36, 31.94, 35.57, 35.93
, 38.42, 61.12, 61.24, 61.71, 61.89, 62.56, 62.
64, 62.69, 69.26, 73.62, 75.69, 75.81, 79.29, 8
2.80, 85.73, 86.04, 86.41, 108.29, 111.23, 111
. 32, 116.46, 116.51, 122.39, 122.98, 123.03, 1
36.55, 136.78, 139.04, 143.11, 143.18, 147.93
, 147.98, 150.30, 150.37, 152.98, 155.39, 155.
59, 163.53, 163.62, 166.00, 166.12, 167.69, 16
7.86, 179.92, 180.04.
TOF/MS _{( ESI} ): calcd _{for C90H149N8O18PNa} [M
+ Na] ⁺ 1684.0625, found 1684.0648.

[Example 3-12]
(4) 2-((3,4,5-tris(octadecyloxy)benzoyl)oxy)acetic acid 5′-O-((2-cyanoethoxy)(5′-O-((2-cyanoethoxy)(5 '-O
-(4,4'-dimethoxytrityl)thymidin-3'-yl)phosphoryl)thymidine-3
Synthesis of '-yl)phosphoryl)-N ² -isobutyryldeoxyguanosin-3'-yl

2-((3,4,5-tris(octadecyloxy)benzoyl)oxy)acetic acid 5′-O-((2-cyanoethoxy)(thymidin-3′-yl)acid according to the method of Example 3-4
phosphoryl)-N ² -isobutyryldeoxyguanosin-3′-yl (2.50 g, 1.
50 mmol) and DMTr-dT-CE phosphoramidite (3.36 g, 4.51 mm
ol) gave the title compound (3.11 g, 89%).
¹ H-NMR (400 MHz, CDCl ₃ ): δ0.88 (t, 9H, J=6.4 Hz)
, 1.05-1.53 (m, 96H), 1.71-1.88 (m, 13H), 2.24-
2.87 (m, 10H), 3.25-3.53 (m, 3H), 3.72-3.81 (m,
7H), 3.95-4.05 (m, 6H), 4.05-4.72 (m, 10H), 4.8
1-4.91 (m, 2H), 5.03-5.24 (m, 2H), 5.55-5.61 (m
, 1H), 5.95-6.43 (m, 3H), 6.82-6.85 (m, 4H), 7.1
8-7.37 (m, 12H), 7.53 (s, 1H), 7.73 (s, 1H), 9.10
-9.31 (m, 2H), 10.05-10.09 (m, 1H), 12.18 (s, 1H)
).
¹³ C-NMR (100 MHz, CDCl ₃ ): δ 11.68, 11.72, 12.33
, 12.37, 14.14, 18.78, 18.88, 19.13, 19.24, 19.
58, 19.65, 19.70, 19.77, 22.70, 26.08, 26.16, 2
9.34, 29.38, 29.46, 29.62, 29.68, 29.74, 29.77
, 30.36, 31.94, 35.84, 35.99, 37.65, 38.95, 55.
29, 61.18, 62.62, 62.77, 62.83, 63.26, 67.10, 6
7.45, 69.27, 73.63, 75.55, 79.91, 82.62, 82.87
, 84.27, 87.31, 108.29, 111.50, 111.80, 113.15
, 113.35, 116.66, 116.74, 116.86, 122.51, 122.
59, 123.07, 127.32, 128.08, 128.13, 129.13, 13
0.13, 134.95, 135.12, 136.32, 138.75, 143.10,
143.99, 147.93, 147.97, 148.04, 150.28, 150.3
7, 150.60, 150.69, 152.98, 155.49, 155.54, 158
. 81, 163.63, 163.68, 163.79, 166.02, 167.69, 1
67.71, 179.73, 179.77.
TOF/MS _{( ESI} ): _{calcd for C124H183N11O27P2Na}
[M+Na] ⁺ 2343.2658, found 2343.2742.

2-((3,4,5-tris(octadecyloxy)benzoyl)oxy)acetic acid 5′-
O-((2-cyanoethoxy)(5'-O-((2-cyanoethoxy)(5'-O-(4
,4′-dimethoxytrityl)thymidin-3′-yl)phosphoryl)-N ² -isobutyryldeoxyguanosin-3′-yl)phosphoryl)thymidin-3′-yl
[Example 3-13]
(1) 2-((3,4,5-tris(octadecyloxy)benzoyl)oxy)acetic acid 5′-O-((2-cyanoethoxy)(N ² -isobutyryldeoxyguanosine-3′-
Synthesis of yl)phosphoryl)thymidin-3′-yl

2-((3,4,5-Tris(octadecyloxy)benzoyl)oxy)thymidin-3′-yl acetate (1.82 g, 1.50 mmol) and N ²
- isobutyryl-DMTr-dG-CE phosphoramidite (2.52 g, 3.00 mm
ol) gave the title compound (2.43 g, 98%).
¹ H-NMR (400 MHz, CDCl ₃ ): δ0.88 (t, 9H, J=7.2 Hz)
, 1.00-1.41 (m, 90H), 1.41-1.54 (m, 6H), 1.71-1
. 85 (m, 6H), 1.92-2.00 (m, 3H), 2.24-2.97 (m, 6H
), 3.68-4.08 (m, 8H), 4.14-4.54 (m, 6H), 4.78-4
. 92 (m, 2H), 5.14-5.78 (m, 2H), 5.95-6.24 (m, 2H
), 7.27-7.43 (m, 3H), 7.87-7.95 (m, 1H), 10.19-
10.49 (m, 2H), 12.24-12.38 (m, 1H).
¹³ C-NMR (100 MHz, CDCl ₃ ): δ12.35, 14.14, 19.01
, 19.06, 19.10, 19.73, 19.80, 19.85, 22.70, 26.
07, 26.14, 29.33, 29.38, 29.45, 29.61, 29.68, 2
9.74, 30.02, 30.36, 31.77, 31.94, 35.80, 35.92
, 36.62, 38.19, 38.48, 61.05, 61.13, 62.26, 62.
44, 62.49, 62.70, 62.75, 66.12, 67.37, 69.21, 7
3.58, 74.51, 80.47, 80.64, 82.33, 82.40, 85.33
, 85.52, 86.12, 86.55, 86.84, 87.31, 108.24, 11
1.24, 112.18, 116.50, 116.83, 121.43, 121.59,
123.05, 123.08, 136.55, 137.73, 138.66, 138.9
9, 143.02, 143.06, 147.75, 147.78, 148.49, 150
. 42, 151.06, 152.96, 152.97, 155.23, 155.27, 1
64.76, 164.83, 165.92, 165.96, 167.73, 167.80
, 180.26, 180.45.
TOF/MS _{( ESI} ): calcd _{for C90H149N8O18PNa} [M
+ Na] ⁺ 1684.0625, found 1684.0643.

[Example 3-14]
(2) 2-((3,4,5-tris(octadecyloxy)benzoyl)oxy)acetic acid 5′-O-((2-cyanoethoxy)(5′-O-((2-cyanoethoxy)(5 '-O
Synthesis of -(4,4′-dimethoxytrityl)thymidin-3′-yl)phosphoryl)-N ² -isobutyryldeoxyguanosin-3′-yl)phosphoryl)thymidin-3′-yl

2-((3,4,5-tris(octadecyloxy)benzoyl)oxy)acetic acid 5′-O-((2-cyanoethoxy)(N ² -isobutyryldeoxyguanosine) according to the method of Example 3-4 -3′-yl)phosphoryl)thymidin-3′-yl (2.20 g, 1.3
2 mmol) and DMTr-dT-CE-phosphoramidite (2.96 g, 3.97 mm
ol) gave the title compound (2.76 g, 90%).
¹ H-NMR (400 MHz, CDCl ₃ ): δ0.88 (t, 9H, J=6.8 Hz)
, 1.13-1.47 (m, 96H), 1.71-2.01 (m, 15H), 2.35-
2.85 (m, 10H), 3.31-3.51 (m, 2H), 3.78 (s, 6H), 3
. 96-4.04 (m, 6H), 4.05-4.59 (m, 10H), 4.84 (s, 2
H), 5.13-5.55 (m, 3H), 6.11-6.42 (m, 3H), 6.83 (
d, 4H, J=8.8Hz), 7.16-7.37 (m, 12H), 7.45-7.55
(m, 1H), 7.72-7.80 (m, 1H), 9.23-10.39 (m, 3H),
12.15-12.21 (m, 1H).
¹³ C-NMR (100 MHz, CDCl ₃ ): δ 11.70, 12.39, 12.45
, 14.14, 18.82, 18.86, 19.04, 19.08, 19.58, 19.
65, 19.74, 19.80, 22.70, 26.08, 26.15, 29.34, 2
9.38, 29.46, 29.61, 29.68, 29.74, 29.77, 30.37
, 31.94, 35.89, 36.13, 36.52, 38.68, 55.29, 61.
12, 62.74, 62.89, 63.22, 63.32, 67.12, 69.23, 7
3.60, 74.76, 79.03, 79.98, 82.49, 84.28, 85.14
, 87.28, 108.26, 111.37, 111.49, 111.69, 111.7
2, 113.14, 113.34, 116.74, 116.84, 116.92, 121
. 89, 121.96, 123.02, 127.08, 127.31, 127.76, 1
27.85, 128.08, 129.13, 130.11, 134.94, 134.99
, 135.03, 135.20, 135.30, 136.93, 138.26, 139.
46, 143.07, 143.96, 144.04, 147.98, 148.02, 14
8.14, 148.23, 150.46, 150.57, 150.62, 152.97,
155.42, 158.60, 158.79, 163.79, 164.43, 164.5
0, 165.96, 167.79, 179.87, 179.95.
TOF/MS _{( ESI} ): _{calcd for C124H183N11O27P2Na}
[M+Na] ⁺ 2343.2658, found 2343.2693.

[Example 3-15]
2-((3,4,5-tris(octadecyloxy)benzoyl)oxy)acetic acid 5′-
Synthesis of O-((2-cyanoethoxy)(5′-O-(4,4′-dimethoxytrityl)-thymidin-3′-yl)phosphorothioyl)-thymidin-3′-yl

2-((3,4,5-tris(octadecyloxy)benzoyl)oxy)thymidin-3′-yl acetate (1.82 g, 1.50 mmol), DMTr-dT-CE phosphoramidite (2.23 g, 3 .00 mmol) in dichloromethane (27 mL) was added with 5-benzylthio-1H-tetrazole (0.58 g, 3.00 mmol), and the mixture was stirred at room temperature for 1.5 hours. ((N,N-dimethylaminomethylidene)amino)-3H
After adding -1,2,4-dithiazoline-3-thione (1.13 g, 4.50 mmol) and pyridine (0.87 g, 9.00 mmol), the mixture was stirred at room temperature for 1.5 hours. Acetonitrile (109 mL) was added dropwise to the reaction solution, and the precipitated solid was filtered. After washing the solid with acetonitrile-dichloromethane mixed solvent, acetonitrile and methanol,
After drying under reduced pressure at °C, the title compound (2.71 g, 96%) was obtained.
¹ H-NMR (400 MHz, CDCl ₃ ): δ0.88 (t, 9H, J=6.8 Hz)
, 1.11-1.53 (m, 93H), 1.72-1.84 (m, 6H), 1.92-1
. 95 (m, 3H), 2.28-2.49 (m, 3H), 2.57-2.66 (m, 2H
), 2.77 (dd, 1H, J = 6.0, 7.2Hz), 3.39-3.50 (m, 2H
), 3.79 (s, 6H), 3.97-4.04 (m, 6H), 4.09-4.40 (m
, 6H), 4.83 (d, 2H, J=6.8Hz), 5.31-5.43 (m, 2H),
6.27-6.32 (m, 1H), 6.35-6.41 (m, 1H), 6.82-6.8
6 (m, 4H), 7.22-7.32 (m, 10H), 7.37-7.39 (m, 2H)
, 7.56 (s, 1H), 8.69-8.91 (m, 2H).
¹³ C-NMR (100 MHz, CDCl ₃ ): δ 11.79, 12.58, 14.14
, 19.31, 19.40, 19.47, 19.55, 22.70, 26.07, 26.
13, 29.32, 29.38, 29.44, 29.60, 29.68, 29.73, 3
0.35, 31.94, 36.78, 39.00, 55.29, 61.06, 62.65
, 62.69, 62.90, 62.94, 63.21, 67.63, 69.19, 73.
56, 75.01, 75.09, 80.18, 82.27, 82.36, 84.40, 8
4.53, 84.91, 85.39, 87.31, 108.25, 111.74, 111
. 90, 113.37, 116.15, 116.39, 123.06, 127.28, 1
28.05, 128.11, 130.09, 134.97, 135.03, 135.06
, 135.30, 143.01, 144.04, 150.22, 150.27, 150.
35, 152.94, 158.78, 163.36, 163.46, 165.86, 16
5.88, 167.71, 167.76.
TOF/MS _{( ESI} ): calcd _{for C107H162N5O19PSNa} [
M+Na] ⁺ 1907.1220, found 1907.1223.

4.2-((3,4,5-tris(octadecyloxy)benzoyl)oxy)acetic acid N
⁴ -benzoyl-5′-O-((2-cyanoethoxy)(5′-O-(4,4′-dimethoxytrityl)-2′-O-methyluridin-3′-yl)phosphoryl)-2′ Synthesis of -O-methylcytidin-3'-yl [Example 4-1]
(1) 2-((3,4,5-tris(octadecyloxy)benzoyl)oxy)acetic acid N ⁴ -benzoyl-5′-O-(4,4′-dimethoxytrityl)-2′-O-methylcytidine Synthesis of -3'-yl

2-((3,4,5-Tris(octadecyloxy)benzoyl)oxy)acetic acid (1.97 g, 2.00 mmol) and N ⁴ -benzoyl-5′- were prepared according to the method of Example 2-1.
O-(4,4'-dimethoxytrityl)-2'-O-methylcytidine (1.59 g, 2.
40 mmol) gave the title compound (2.96 g, 91%).
¹ H-NMR (400 MHz, CDCl ₃ ): δ0.88 (t, 9H, J=6.8 Hz)
, 1.10-1.38 (m, 84H), 1.42-1.50 (m, 6H), 1.70-1
. 85 (m, 6H), 3.48 (d, 1H, J=11.6Hz), 3.62 (s, 3H)
, 3.69(d, 1H, J=11.2Hz), 3.71(s, 1H), 3.83(s, 5
H), 3.97-4.04 (m, 6H), 4.18 (d, 1H, J=4.8Hz), 4.
36 (d, 1H, J = 8.4Hz), 4.80-4.89 (m, 2H), 5.29 (dd
, 1H, J=4.8 Hz, 8.4 Hz), 6.11 (d, 1 H, J=1.6 Hz), 6.
90 (dd, 4H, J=2.0, 8.8Hz), 7.27-7.42 (m, 11H), 7
. 52 (t, 2H, J=7.6Hz), 7.62 (t, 1H, J=7.6Hz), 7.8
9 (d, 2H, J = 7.6Hz), 8.55 (d, 1H, J = 6.4Hz), 8.61-
8.72 (brs, 1H).
¹³ C-NMR (100 MHz, CDCl ₃ ): δ 14.14, 22.70, 26.07
, 26.09, 26.11, 29.31, 29.38, 29.43, 29.59, 29.
66, 29.68, 29.73, 30.34, 31.94, 52.31, 55.24, 5
9.15, 60.47, 60.72, 61.06, 69.14, 70.07, 73.53
, 80.42, 82.21, 87.49, 88.91, 108.22, 113.43, 1
23.33, 123.50, 127.27, 127.49, 128.17, 129.06
, 130.06, 130.08, 133.17, 135.07, 135.22, 142.
85, 143.86, 152.87, 152.90, 158.75, 165.64, 16
5.84, 167.17, 168.42.
TOF/MS ( _ESI ): calcd _{for C101H151N3O14Na [} M+
Na] ⁺ 1653.1094, found 1653.1171.

[Example 4-2]
(2) Synthesis of 2-((3,4,5-tris(octadecyloxy)benzoyl)oxy)acetic acid N ⁴ -benzoyl-2′-O-methylcytidin-3′-yl

2-((3,4,5-Tris(octadecyloxy)benzoyl)oxy)acetic acid N ⁴ -benzoyl-5′-O-(4,4′-dimethoxytrityl)-2 according to the method of Example 2-2 '-O-Methylcytidin-3'-yl (2.80 g, 1.72 mmol) gave the title compound (2.26 g, 99%).
¹ H-NMR (400 MHz, CDCl ₃ ): δ0.88 (t, 9H, J=6.8 Hz)
, 1.10-1.55 (m, 90H), 1.70-1.85 (m, 6H), 3.51 (s
, 3H), 3.79-3.83 (m, 2H), 3.95-4.08 (m, 7H), 4.3
4 (d, 1H, J = 4.8Hz), 4.50 (t, 1H, J = 4.8Hz), 4.83-
4.90 (m, 2H), 5.44 (t, 1H, J=4.8Hz), 5.73 (d, 1H,
J = 4.4 Hz), 7.29-7.30 (m, 2H), 7.47-7.66 (m, 3H)
, 7.90 (d, 2H, J=7.6Hz), 8.16 (d, 1H, J=7.6Hz), 8
. 75-9.00 (brs, 1H).
¹³ C-NMR (100 MHz, CDCl ₃ ): δ 14.14, 22.71, 26.07
, 26.11, 29.30, 29.38, 29.42, 29.59, 29.66, 29.
68, 29.73, 30.34, 31.94, 52.32, 59.19, 60.97, 6
1.07, 61.24, 69.15, 69.20, 71.18, 73.56, 80.58
, 83.43, 92.64, 108.28, 123.20, 123.50, 127.61
, 129.09, 132.80, 133.36, 142.99, 147.07, 152.
87, 152.93, 162.63, 165.85, 167.66, 168.43.
TOF/MS ( _ESI ): _calcd for _{C80H133N3O12Na} [ _M +N
a] ⁺ 1350.9787, found 1350.9855.

[Example 4-3]
(3) 2-((3,4,5-tris(octadecyloxy)benzoyl)oxy)acetic acid N ⁴ -benzoyl-5′-O-((2-cyanoethoxy)(5′-O-(4,4 '-dimethoxytrityl)-2'-O-methyluridin-3'-yl)phosphoryl)-2'-O-
Synthesis of methylcytidin-3'-yl

2-((3,4,5-Tris(octadecyloxy)benzoyl)oxy)acetic acid N ⁴ -benzoyl-2′-O-methylcytidin-3′-yl (2
. 10 g, 1.58 mmol) and 5′-O-DMTr-2′-O-methyluridine-CE
A crude product (2.80 g) was obtained from the phosphoramidite (2.40 g, 3.16 mmol).
The crude product was purified by column chromatography (neutral silica gel, developing solvent: dichloromethane-methanol) to obtain the title compound (2.21 g, 70%).
¹ H-NMR (400 MHz, CDCl ₃ ): δ0.88 (t, 9H, J=6.8 Hz)
, 1.10-1.53 (m, 90H), 1.71-1.83 (m, 6H), 2.50-2
. 74 (m, 1H), 2.79 (t, 1H, J=6.4Hz), 3.46-3.76 (m
, 8H), 3.77 (s, 3H), 3.79 (s, 3H), 3.97-4.04 (m, 6
H), 4.11-4.58 (m, 7H), 4.82-4.95 (m, 2H), 5.13-
5.30 (m, 3H), 5.86-6.08 (m, 2H), 6.82-6.88 (m, 4
H), 7.23-7.36 (m, 11H), 7.51 (t, 2H, 8.0Hz), 7.6
1 (t, 1H, J=7.2Hz), 7.82-8.05 (m, 4H), 8.69-8.9
6(m, 2H).
¹³ C-NMR (100 MHz, CDCl ₃ ): δ 14.14, 19.52, 19.58
, 22.70, 26.07, 26.13, 29.33, 29.38, 29.45, 29.
60, 29.68, 29.73, 30.35, 31.94, 55.25, 55.30, 5
8.84, 59.27, 60.82, 61.14, 62.35, 62.40, 62.68
, 62.73, 65.55, 66.16, 69.19, 69.90, 70.16, 73.
56, 74.28, 74.38, 79.38, 79.46, 79.52, 79.60, 8
1.32, 81.38, 81.52, 82.19, 86.71, 86.87, 87.46
, 90.07, 90.64, 102.58, 102.62, 108.24, 113.34
, 113.36, 116.36, 116.43, 123.19, 127.39, 127.
44, 127.64, 128.10, 128.12, 128.26, 128.29, 12
9.04, 130.21, 130.27, 130.30, 133.28, 134.64,
134.73, 139.53, 139.75, 142.96, 143.82, 143.9
0, 150.02, 150.07, 152.93, 158.81, 158.84, 158
. 86, 162.70, 162.73, 165.77, 165.81, 167.22, 1
67.26.
TOF/MS ( _ESI ): calcd _{for C114H167N6O22PNa} [ _M
+ Na] ⁺ 2026.1769, found 2026.1764.

2-((3,4,5-tris(octadecyloxy)benzoyl)oxy)acetic acid 5′-
O-((2-cyanoethoxy)(5'-O-(4,4'-dimethoxytrityl)-2'-
Synthesis of fluorodeoxyuridin-3′-yl)phosphoryl)-2′-fluoro-N ² -isobutyryldeoxyguanosin-3′-yl
[Example 4-4]
(1) 2-((3,4,5-tris(octadecyloxy)benzoyl)oxy)acetic acid 5′-O-(4,4′-dimethoxytrityl)-2′-fluoro-N ² -isobutyryldeoxy Synthesis of guanosin-3'-yl

2-((3,4,5-Tris(octadecyloxy)benzoyl)oxy)acetic acid (1.97 g, 2.00 mmol) and 5′-O-(4,4′- dimethoxytrityl)-2′-fluoro-N ² -isobutyryldeoxyguanosine (1.5
8 g, 2.40 mmol) gave the title compound (3.13 g, 96%).
¹ H-NMR (400 MHz, CDCl ₃ ): δ0.81 (d, 3H, J=6.8 Hz)
, 0.88 (t, 9H, J = 6.8Hz), 0.94 (d, 3H, J = 7.2Hz), 1
. 10-1.53 (m, 90H), 1.70-1.87 (m, 6H), 1.93 (sep
t, 1H, J = 6.8 Hz), 3.17 (d, 1H, J = 11.2 Hz), 3.59 (d
, 1H, J = 11.2 Hz), 3.77 (s, 6H), 3.97-4.05 (m, 6H)
, 4.32-4.34 (m, 1H), 4.85 (d, 1H, J = 16.0 Hz), 4.9
2 (d, 1H, J=16.0Hz), 5.92-6.07 (m, 2H), 6.18-6.
24 (m, 1H), 6.78 (dd, 4H, J=5.2, 8.8Hz), 7.16-7.
29 (m, 7H), 7.30 (s, 2H), 7.36 (d, 2H, J=8.0Hz), 7
. 79 (s, 1H), 8.01 (s, 1H), 11.92 (s, 1H).
¹³ C-NMR (100 MHz, CDCl ₃ ): δ 14.14, 18.39, 18.47
, 18.88, 18.91, 22.70, 25.61, 26.06, 26.11, 29.
30, 29.38, 29.42, 29.59, 29.66, 29.68, 29.73, 3
0.34, 31.94, 36.25, 36.43, 55.24, 55.25, 60.68
, 60.84, 61.24, 67.98, 69.25, 70.85, 71.00, 73.
61, 79.97, 81.44, 83.15, 86.36, 86.47, 86.67, 9
0.96, 108.26, 113.15, 113.22, 122.23, 123.17,
127.08, 127.13, 127.76, 127.85, 127.89, 127.9
8, 129.13, 129.85, 129.92, 135.20, 135.66, 138
. 43, 138.70, 139.43, 143.05, 144.47, 147.30, 1
47.34, 147.47, 147.90, 152.93, 152.96, 155.04
, 155.31, 158.62, 158.69, 165.65, 165.89, 167.
23, 167.60, 178.18, 178.76.
TOF/MS ( _ESI ): _calcd for _{C98H150N5O13F} [M+ _H ]
⁺ 1623.1136, found 1623.1160.

[Example 4-5]
(2) Synthesis of 2-((3,4,5-tris(octadecyloxy)benzoyl)oxy)acetic acid 2′-fluoro-N ² -isobutyryldeoxyguanosin-3′-yl

2-((3,4,5-tris(octadecyloxy)benzoyl)oxy)acetic acid 5′-O-(4,4′-dimethoxytrityl)-2′-fluoro-N according to the method of Example 2-2 ² -isobutyryldeoxyguanosin-3′-yl (3.00 g, 1.85 mmol
) gave the title compound (2.40 g, 98%).
¹ H-NMR (400 MHz, CDCl ₃ ): δ0.88 (t, 9H, J=6.8 Hz)
, 1.10-1.55 (m, 96H), 1.70-1.85 (m, 6H), 2.71 (s
ept, 1H, J = 6.8Hz), 3.37-3.72 (brs, 2H), 3.80 (d
, 1H, J = 11.6 Hz), 3.99-4.05 (m, 7H), 4.34 (d, 1H,
J = 4.0 Hz), 4.94 (s, 2H), 5.69 (dt, 1H, J = 4.4, 52.
0 Hz), 5.78-5.84 (m, 1H), 5.97 (dd, 1H, J = 4.0, 16
. 0 Hz), 7.31 (s, 2H), 7.90 (s, 1H), 9.22 (s, 1H), 1
2.15(s, 1H).
¹³ C-NMR (100 MHz, CDCl ₃ ): δ 14.14, 18.85, 18.95
, 22.70, 26.06, 26.11, 29.30, 29.38, 29.43, 29.
60, 29.68, 29.74, 30.33, 31.94, 36.32, 60.86, 6
9.24, 71.25, 71.39, 73.60, 82.89, 87.22, 87.55
, 89.60, 91.55, 108.27, 121.56, 123.15, 138.45
, 142.99, 147.29, 148.08, 152.91, 155.03, 165.
91, 167.26, 179.15.
TOF/MS ₍ ESI): _{calcd for C77H132N5O11FNa} [M+
Na] ⁺ 1344.9805, found 1344.9834.

[Example 4-6]
(3) 2-((3,4,5-tris(octadecyloxy)benzoyl)oxy)acetic acid 5′-O-((2-cyanoethoxy)(5′-O-(4,4′-dimethoxytrityl) -
2'-fluorodeoxyuridin-3'-yl)phosphoryl)-2'-fluoro-N ² -
Synthesis of isobutyryldeoxyguanosin-3′-yl

2-((3,4,5-tris(octadecyloxy)benzoyl)oxy)acetic acid 2′-fluoro-N ² -isobutyryldeoxyguanosine-3′ according to the method of Example 3-4
-yl (2.20 g, 1.66 mmol) and 5′-O-DMTr-2′-fluoro-deoxyuridine-CE phosphoramidite (2.49 g, 3.33 mmol) to give the title compound (3.07 g, 93 mmol). %) was obtained.
¹ H-NMR (400 MHz, CDCl ₃ ): 0.88 (t, 9 H, J=7.2 Hz),
0.97-1.53 (m, 96H), 1.71-1.83 (m, 6H), 2.56-2.
75 (m, 3H), 3.37-3.66 (m, 2H), 3.74-3.79 (m, 6H)
, 3.97-4.55 (m, 12H), 4.78-5.08 (m, 2H), 5.08-5
. 50 (m, 3H), 5.70-6.01 (m, 4H), 6.80-6.84 (m, 4H
), 7.17-7.40 (m, 11H), 7.57-7.73 (m, 2H), 8.94-
9.07 (m, 1H), 10.00 (s, 1H), 12.18-12.26 (m, 1H)
.
δ ¹³ C-NMR (100 MHz, CDCl ₃ ): δ 14.14, 18.51, 18.7
6, 18.92, 19.20, 19.55, 19.62, 22.70, 26.07, 26
. 14, 29.33, 29.38, 29.45, 29.60, 29.68, 29.74,
30.36, 31.94, 35.90, 35.99, 55.23, 55.30, 60.7
0, 60.88, 62.85, 69.26, 73.61, 79.54, 79.63, 87
. 17, 87.21, 102.93, 108.31, 113.31, 116.34, 11
6.52, 122.55, 122.61, 122.95, 122.98, 127.25,
127.31, 128.07, 128.10, 130.14, 134.79, 134.8
8, 134.92, 138.65, 140.55, 143.15, 143.19, 143
. 92, 144.02, 147.71, 147.74, 148.30, 148.43, 1
49.68, 149.98, 152.98, 152.99, 155.30, 155.48
, 158.74, 158.79, 162.67, 165.89, 166.00, 167.
20, 179.68.
TOF/MS ( _ESI ) _{: calcd for C110H163N8O20F2PNa}
[M+Na] ⁺ 2008.1587, found 2008.1593.

2-((3,4,5-tris(octadecyloxy)benzoyl)oxy)acetic acid 5′-
O-((2-cyanoethoxy)(5'-O-(4,4'-dimethoxytrityl)-2'-
O-(tert-butyldimethylsilyl)uridin-3'-yl)phosphoryl)-2'-
Synthesis of O-(tert-butyldimethylsilyl)uridin-3′-yl [Example 4-7]
(1) 2-((3,4,5-tris(octadecyloxy)benzoyl)oxy)acetic acid 5′-O-(4,4′-dimethoxytrityl)-2′-O-(tert-butyldimethylsilyl) Synthesis of uridin-3'-yl

2-((3,4,5-tris(octadecyloxy)benzoyl)oxy)acetic acid (2.
46 g, 2.50 mmol) and 5′-O-(4,4′-dimethoxytrityl)-2′-O
To a suspension of -(tert-butyldimethylsilyl)uridine (1.98 g, 3.00 mmol) in dichloromethane (25 mL) was added 4-dimethylaminopyridine (0.03 g, 0.2
5 mmol) and 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride (0.58 g, 3.00 mmol) were added, followed by stirring at room temperature for 5 hours. Acetonitrile (148 mL) was added dropwise to the reaction solution, and the precipitated solid was filtered. After washing the solid with a mixed solvent of acetonitrile and dichloromethane, it was dried under reduced pressure at 50° C. to give a crude product (2.89 g
). The crude product was purified by column chromatography (neutral silica gel, developing solvent: dichloromethane-THF) to give the title compound (2.76 g, 68%) as a white solid.
¹ H-NMR (400 MHz, CDCl ₃ ): δ 0.08 (s, 3H), 0.11 (s,
3H), 0.81-0.92 (m, 18H), 1.04-1.53 (m, 90H), 1.
70-1.84 (m, 6H), 3.49 (dd, 2H, J = 10.8, 27.6Hz),
3.79 (s, 6H), 3.79-4.04 (m, 6H), 4.26-4.27 (m, 1
H), 4.56 (t, 1H, J = 5.2Hz), 4.81 (d, 1H, J = 16.0Hz
), 4.96 (d, 1H, 16.4Hz), 5.30 (d, 1H, J = 8.0Hz), 5
. 47 (t, 1H, J=4.0Hz), 6.02 (d, 1H, J=5.6Hz), 6.8
5 (d, 4H, J=8.4Hz), 7.23-7.37 (m, 11H), 7.85 (d,
1H, J=8.4Hz), 8.37(s, 1H).
¹³ C-NMR (100 MHz, CDCl ₃ ): δ -5.22, -5.09, 14.14
, 17.87, 22.71, 25.51, 26.07, 26.11, 29.30, 29.
38, 29.43, 29.59, 29.67, 29.74, 30.34, 31.94, 5
5.25, 60.75, 62.47, 69.14, 73.36, 73.54, 74.40
, 81.20, 87.58, 88.03, 102.56, 108.25, 113.40,
113.43, 123.33, 127.33, 128.03, 128.14, 130.0
3, 130.17, 134.71, 134.83, 140.00, 142.87, 144
. 10, 150.14, 152.89, 158.78, 158.83, 162.59, 1
65.67, 167.10.
TOF/MS ₍ ESI): _{calcd for C99H158N2O14SiNa} [M
+ Na] ⁺ 1650.1380, found 1650.1455.

[Example 4-8]
(2) Synthesis of 2-((3,4,5-tris(octadecyloxy)benzoyl)oxy)acetic acid 2′-O-(tert-butyldimethylsilyl)uridin-3′-yl

2-((3,4,5-Tris(octadecyloxy)benzoyl)oxy)acetic acid 5′-O-(4,4′-dimethoxytrityl)-2′-O-( t
ert-butyldimethylsilyl)uridin-3′-yl (2.60 g, 1.60 mmol
) gave the title compound (2.05 g, 97%).
¹ H-NMR (400 MHz, CDCl ₃ ): δ 0.04 (s, 3H), 0.07 (s,
3H), 0.81-0.92 (m, 18H), 1.11-1.53 (m, 90H), 1.
70-1.85 (m, 6H), 2.95-3.10 (brs, 1H), 3.77-3.8
2 (m, 1H), 3.93-4.04 (m, 7H), 4.25-4.28 (m, 1H),
4.67 (t, 1H, J=5.2Hz), 4.81 (d, 1H, 16.0Hz), 4.9
6 (d, 1H, 16.0Hz), 5.34 (dd, 1H, J = 3.2, 4.8Hz), 5
. 60 (d, 1H, J = 5.6Hz), 5.76 (d, 1H, J = 8.0Hz), 7.2
9 (s, 2H), 7.63 (d, 1H, J=8.0 Hz), 8.71 (s, 1H).
¹³ C-NMR (100 MHz, CDCl ₃ ): δ -5.22, -5.19, 14.14
, 17.88, 22.71, 25.52, 26.07, 26.10, 29.30, 29.
38, 29.42, 29.59, 29.66, 29.68, 29.73, 30.34, 3
1.94, 60.84, 61.84, 69.20, 72.78, 73.24, 73.57
, 82.99, 92.46, 102.70, 108.32, 123.24, 141.88
, 142.99, 150.17, 152.92, 162.72, 165.78, 167.
38.
TOF/MS _{( ESI} ): calcd _{for C78H140N2O12SiNa} [M
+ Na] ⁺ 1348.0073, found 1348.0115.

[Example 4-9]
(3) 2-((3,4,5-tris(octadecyloxy)benzoyl)oxy)acetic acid 5′-O-((2-cyanoethoxy)(5′-O-(4,4′-dimethoxytrityl) -
2′-O-(tert-butyldimethylsilyl)uridin-3′-yl)phosphoryl)-
Synthesis of 2′-O-(tert-butyldimethylsilyl)uridin-3′-yl

2-((3,4,5-tris(octadecyloxy)benzoyl)oxy)acetic acid 2′-O-(tert-butyldimethylsilyl)uridine-3′ according to the method of Example 3-4
-yl (1.95 g, 1.47 mmol) and 5′-O-DMTr-2′-O-(tert
-butyldimethylsilyl)uridine-CE phosphoramidite (2.53g, 2.94m)
mol) gave the title compound (2.84 g, 92%).
¹ H-NMR (400 MHz, CDCl ₃ ): δ 0.03-0.13 (m, 12H), 0
. 86-0.90 (m, 27H), 1.02-1.53 (m, 90H), 1.68-1.
84 (m, 6H), 2.45-2.69 (m, 1H), 2.73 (t, 1H, J = 6.0
Hz), 3.48 (d, 1H, J=10.8Hz), 3.60-3.68 (m, 1H),
3.76-3.84 (m, 6H), 3.97-4.04 (m, 6H), 4.10-4.5
8 (m, 8H), 4.73-4.84 (m, 1H), 4.90-5.03 (m, 2H),
5.20-5.25 (m, 2H), 5.67-5.78 (m, 2H), 5.95-6.0
3 (m, 1H), 6.80-6.89 (m, 4H), 7.21-7.44 (m, 12H)
, 7.84 (d, 1H, J=8.0 Hz), 8.72-8.83 (m, 2H).
¹³ C-NMR (100 MHz, CDCl ₃ ): δ -5.27, -5.24, -5.07
, -4.94, -4.92, 14.14, 17.86, 18.06, 19.50, 19.
57, 19.65, 22.70, 25.50, 25.55, 25.60, 26.07, 2
6.13, 29.32, 29.38, 29.44, 29.60, 29.68, 29.74
, 30.35, 31.94, 55.27, 55.30, 60.69, 61.96, 62.
37, 62.42, 62.58, 62.63, 69.19, 71.39, 71.60, 7
2.86, 73.00, 73.57, 74.47, 79.43, 79.51, 81.75
, 87.65, 87.68, 87.99, 88.18, 90.56, 91.22, 102
. 54, 102.64, 103.05, 103.13, 108.28, 113.39, 1
16.31, 116.40, 123.21, 127.43, 128.13, 128.21
, 128.25, 130.25, 130.27, 130.32, 134.56, 134.
65, 134.70, 134.73, 139.84, 140.05, 140.15, 14
2.97, 143.01, 143.92, 143.97, 149.95, 150.19,
150.40, 152.92, 158.83, 158.87, 162.53, 162.5
8, 162.69, 165.64, 165.68, 167.05, 167.12.
TOF/MS _{( ESI} ): _{calcd for C117H186N5O22Si2PN}
a[M+Na] ⁺ 2123.2763, found 2123.2769.

5. Synthesis of 3-((3,4,5-tris(octadecyloxy)benzoyl)oxy)propionic acid [Example 5-1]
(1) Synthesis of tert-butyl 3-((3,4,5-tris(octadecyloxy)benzoyl)oxy)propionate

3,4,5-tris(octadecyloxy)benzoic acid (2.78 g, 3.0 mmol)
, 4-dimethylaminopyridine (0.04 g, 0.3 mmol), tert-butyl 3-hydroxypropionate (4.39 g, 30.0 mmol), triethylamine (0.4
6 g, 4.5 mmol) in dichloromethane (60 mL) was added with 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride (0.86 g, 4.5 mmol) and stirred at room temperature for 20 hours. bottom. Acetonitrile (180 mL) was added dropwise to the reaction solution, and the precipitated solid was filtered. The solid was washed with an acetonitrile-dichloromethane mixed solvent and then dried under reduced pressure at 50° C. to obtain a crude product (3.07 g). The crude product was purified by column chromatography (neutral silica gel, developing solvent: dichloromethane) to give the title compound (1.90 g, 60%) as a white solid.
¹ H-NMR (400 MHz, CDCl ₃ ): δ0.88 (t, 9H, J=7.0 Hz)
, 1.15-1.55 (m, 99H), 1.70-1.85 (m, 6H), 2.68 (t
, 2H, J = 6.4 Hz), 3.95-4.05 (m, 6H), 4.53 (t, 2H, J
= 6.4Hz), 7.23(s, 2H).
¹³ C-NMR (100 MHz, CDCl ₃ ): δ 14.08, 22.68, 26.08
, 28.09, 29.35, 29.40, 29.57, 29.65, 29.71, 30.
35, 31.93, 35.42, 60.72, 69.25, 73.51, 80.97, 1
08.19, 124.57, 152.84, 166.14, 169.93.
TOF/MS (ESI): _{calcd for C68H126O7Na} [M+Na] ⁺
1077.9401, found 1077.9427.

[Example 5-2]
(2) Synthesis of 3-((3,4,5-tris(octadecyloxy)benzoyl)oxy)propionic acid

To a solution of tert-butyl 3-((3,4,5-tris(octadecyloxy)benzoyl)oxy)propionate (2.75 g, 2.6 mmol) in dichloromethane (52 mL) was added methanesulfonic acid (3.75 g, 39. 0 mmol) was added, and the mixture was stirred at room temperature for 20 hours. Acetonitrile (156 mL) was added dropwise to the reaction solution, and the precipitated solid was filtered. The solid was washed with an acetonitrile-dichloromethane mixed solvent and acetonitrile, and dried under reduced pressure at 50° C. to give the title compound (2.54 g, 98%) as a white solid.
¹ H-NMR (400 MHz, CDCl ₃ ): δ0.88 (t, 9H, J=7.0 Hz)
, 1.15-1.55 (m, 90H), 1.70-1.85 (m, 6H), 2.85 (t
, 2H, J = 6.6 Hz), 3.95-4.10 (m, 6H), 4.57 (t, 2H, J
= 6.4Hz), 7.23(s, 2H).
¹³ C-NMR (100 MHz, CDCl ₃ ): δ14.10, 22.71, 26.10
, 26.13, 29.38, 29.45, 29.61, 29.68, 29.75, 30.
38, 31.96, 33.54, 59.99, 69.34, 73.57, 108.35,
124.32, 142.88, 152.91, 166.17.
TOF/MS ( _ESI ): calcd _{for C64H118O7Na} [M+Na]
⁺ 1021.8775, found 1021.8817.

Synthesis of 2-(N-methyl-3,4,5-tris(octadecyloxy)benzamido)acetic acid [Example 5-3]
(1) 2-(N-methyl-3,4,5-tris(octadecyloxy)benzamide)
Synthesis of tert-butyl acetate

3,4,5-tris(octadecyloxy)benzoic acid (2.78 g, 3.0 mmol)
, 4-dimethylaminopyridine (0.04 g, 0.3 mmol), sarcosine tert-
Butyl hydrochloride (2.73 g, 15.0 mmol), triethylamine (1.52 g, 15
. 0 mmol) in dichloromethane (60 mL) was added with 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride (0.86 g, 4.5 mmol),
Stir at room temperature for 20 hours. Acetonitrile (180 mL) was added dropwise to the reaction solution, and the precipitated solid was filtered. After washing the solid with an acetonitrile-dichloromethane mixed solvent,
It was dried under reduced pressure at 50° C. to obtain a crude product (3.07 g). The crude product was purified by column chromatography (neutral silica gel, developing solvent: dichloromethane-THF) to give the title compound (2.66 g, 84%) as a white solid.
¹ H-NMR (400 MHz, CDCl ₃ ): δ0.88 (t, 9H, J=7.0 Hz)
, 1.15-1.55 (m, 99H), 1.70-1.85 (m, 6H), 3.04 (s
, 3H), 3.80-4.20 (m, 8H), 6.63 (s, 2H).
¹³ C-NMR (100 MHz, CDCl ₃ ): δ 14.01, 22.66, 26.13
, 26.15, 28.16, 29.35, 29.42, 29.48, 29.65, 29.
71, 30.39, 31.93, 69.51, 73.52, 82.02, 106.19,
130.61, 139.93, 153.25, 168.33, 172.01.
TOF/MS ( _ESI ): calcd _{for C68H127NO6Na} [M+Na]
⁺ 1076.9561, found 1076.9591.

[Example 5-4]
(2) 2-(N-methyl-3,4,5-tris(octadecyloxy)benzamide)
Synthesis of acetic acid

The title compound (2.38 g) was obtained from tert-butyl 2-(N-methyl-3,4,5-tris(octadecyloxy)benzamido)acetate (2.53 g, 2.4 mmol) according to the method of Example 5-2. , 99%) as a white solid.
¹ H-NMR (400 MHz, CDCl ₃ ): δ0.88 (t, 9H, J=7.0 Hz)
, 1.15-1.55 (m, 90H), 1.70-1.85 (m, 6H), 3.11 (s
, 3H), 3.85-4.35 (m, 8H), 6.15-6.30 (brs, 1H), 6
. 66(s, 2H).
¹³ C-NMR (100 MHz, CDCl ₃ ): δ 14.05, 22.67, 26.12
, 29.36, 29.43, 29.61, 29.66, 29.72, 29.98, 30.
36, 31.93, 39.20, 50.02, 69.49, 73.56, 106.47,
129.16, 140.27, 152.97, 153.24, 172.11, 172.9
2.
TOF/MS (ESI): calcd for C ₆₄ H ₁₁₈ NO ₆ [MH] ⁻ 9
96.8959, found 996.8947.

Synthesis of 3-(3,4,5-tris(octadecyloxy)benzamido)propionic acid [Example 5-5]
(1) Synthesis of tert-butyl 3-(3,4,5-tris(octadecyloxy)benzamido)propionate

3,4,5-tris(octadecyloxy)benzoic acid (2.78 g, 3.0 mmol)
, 4-dimethylaminopyridine (0.04 g, 0.3 mmol), β-alanine tert
- butyl ester hydrochloride (2.73 g, 15.0 mmol), triethylamine (1.5
2 g, 15.0 mmol) in dichloromethane (60 mL) was added with 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride (0.86 g, 4.5 mmol). Stirred for an hour. Acetonitrile (180 mL) was added dropwise to the reaction solution, and the precipitated solid was filtered. The solid was washed with an acetonitrile-dichloromethane mixed solvent and then dried under reduced pressure at 50° C. to obtain a crude product (3.12 g). The crude product was purified by column chromatography (neutral silica gel, developing solvent: dichloromethane-THF) to give the title compound (2.75 g, 87%) as a white solid.
¹ H-NMR (400 MHz, CDCl ₃ ): δ0.88 (t, 9H, J=7.0 Hz)
, 1.15-1.55 (m, 99H), 1.70-1.85 (m, 6H), 2.55 (t
, 2H, J = 5.8 Hz), 3.66 (q, 2H, J = 6.0 Hz), 3.95-4.0
5 (m, 6H), 6.72 (t, 1H, J=6.0Hz), 6.94 (s, 2H).
¹³ C-NMR (100 MHz, CDCl ₃ ): δ 14.09, 22.71, 26.11
, 28.16, 29.38, 29.43, 29.61, 29.68, 29.74, 30.
36, 31.95, 35.18, 35.58, 69.45, 73.53, 81.19, 1
05.86, 129.57, 141.32, 153.14, 167.20, 172.27
.
TOF/MS ( _ESI ): calcd _{for C68H127NO6Na} [M+Na]
⁺ 1076.9561, found 1076.9579.

[Example 5-6]
(2) Synthesis of 3-(3,4,5-tris(octadecyloxy)benzamido)propionic acid

The title compound (2.52 g, 101%) was prepared from tert-butyl 3-(3,4,5-tris(octadecyloxy)benzamido)propionate (2.64 g, 2.5 mmol) according to the method of Example 5-2. ) as a white solid.
¹ H-NMR (400 MHz, CDCl ₃ ): δ0.88 (t, 9H, J=7.0 Hz)
, 1.15-1.55 (m, 90H), 1.65-1.85 (m, 6H), 2.74 (t
, 2H, J = 5.8 Hz), 3.72 (t, 2H, J = 5.8 Hz), 3.90-4.0
5 (m, 6H), 6.95 (s, 2H).
¹³ C-NMR (100 MHz, CDCl ₃ ): δ 14.06, 22.71, 26.16
, 26.19, 29.40, 29.50, 29.53, 29.65, 29.71, 29.
77, 30.44, 31.98, 33.68, 35.85, 39.37, 69.77, 7
3.67, 106.51, 128.33, 142.29, 153.35, 168.43,
175.46.
TOF/MS (ESI): calcd for C ₆₄ H ₁₁₈ NO ₆ [MH] ⁻ 9
96.8959, found 996.8912.

3-((3,4,5-tris(octadecyloxy)benzoyl)oxy)propionic acid 5′-O-((2-cyanoethoxy)(5′-O-(4,4′-dimethoxytrityl)
Synthesis of thymidin-3′-yl)phosphoryl)thymidin-3′-yl [Example 5-7]
(1) Synthesis of thymidin-3′-yl 3-((3,4,5-tris(octadecyloxy)benzoyl)oxy)propionate

3-((3,4,5-tris(octadecyloxy)benzoyl)oxy)propionic acid (2.40 g, 2.40 mmol), 5′-O-(4,4′-dimethoxytrityl) thymidine (1.57 g) , 2.88 mmol), 1-methylimidazole (0.99 g, 12
. 00 mmol) of COMU (2.06 g, 4.80 mmol) in THF (24 mL).
l) was added and stirred at room temperature for 6.5 hours. Furthermore, COMU (0.21 g, 0.48
mmol) was added, and the mixture was stirred at room temperature for 17.5 hours. Acetonitrile (12
0 mL) was added dropwise, and the precipitated solid was filtered. The solid was washed with an acetonitrile-THF mixed solvent and then dried under reduced pressure at 50° C. to obtain a crude product (2.76 g). The crude product was used for the next reaction without purification.

Pyrrole (0.57 g, 8.5 g) was added to a dichloromethane solution (68 mL) of the crude product (2.60 g).
0 mmol) and trifluoroacetic acid (0.24 g, 2.13 mmol) were added, and the mixture was stirred at room temperature for 3.5 hours. After adding trifluoroacetic acid (0.05 g, 0.43 mmol), the mixture was stirred at room temperature for 1.5 hours. Acetone (20 mL) and acetonitrile (61 mL) were added dropwise to the reaction solution, and the precipitated solid was filtered. The solid was washed with a mixed solvent of acetonitrile-acetone-dichloromethane and acetonitrile, and dried under reduced pressure at 50° C. to obtain a crude product (2.44 g). The title compound (0.52 g, 1
8%) was obtained as a white solid.
¹ H-NMR (400 MHz, CDCl ₃ ): δ0.88 (t, 9H, J=6.8 Hz)
, 1.15-1.55 (m, 90H), 1.65-1.85 (m, 7H), 1.92 (s
, 3H), 2.30-2.40 (m, 1H), 2.45-2.55 (m, 1H), 2.6
0-2.70 (m, 1H), 2.80-2.90 (m, 2H), 3.85-3.95 (m
, 2H), 3.95-4.05 (m, 6H), 4.05-4.10 (m, 1H), 4.5
5-. 65 (m, 2H), 5.40-5.55 (m, 1H), 6.10-6.20 (m,
1H), 7.22 (s, 2H), 7.41 (s, 1H), 8.58-8.64 (m, 1H
).
¹³ C-NMR (100 MHz, CDCl ₃ ): δ12.57, 14.14, 22.71
, 26.07, 26.11, 29.33, 29.38, 29.43, 29.59, 29.
67, 29.74, 30.02, 30.34, 31.94, 34.16, 36.90, 6
0.05, 62.58, 69.24, 73.54, 75.25, 85.10, 86.67
, 108.06, 111.42, 124.14, 136.56, 142.69, 150.
26, 152.87, 163.40, 166.10, 170.43.
TOF/MS ( _ESI ): _{calcd for C74H130N2O11Na} [M+N
a] ⁺ 1245.9572, found 1245.9548.

[Example 5-8]
(2) 3-((3,4,5-tris(octadecyloxy)benzoyl)oxy)propionate 5′-O-((2-cyanoethoxy)(5′-O-(4,4′-dimethoxytrityl) ) thymidin-3′-yl)phosphoryl)thymidin-3′-yl

3-((3,4,5-tris((octadecyloxy)
benzoyl)oxy)thymidin-3′-yl propionate (1.50 g, 1.23 mmo
l) and DMTr-dT-CE phosphoramidite (1.83 g, 2.45 mmol) gave the title compound (2.22 g, 96%) as a white solid.
¹ H-NMR (400 MHz, CDCl ₃ ): δ0.88 (t, 9H, J=6.8 Hz)
, 1.10-1.38 (m, 84H), 1.41-1.50 (m, 9H), 1.70-1
. 83 (m, 7H), 1.91 (s, 3H), 2.30-2.46 (m, 3H), 2.6
2-2.68 (m, 2H), 2.75 (t, 1H, J=6.0Hz), 2.84 (dd,
2H, J = 4.8, 6.4Hz), 3.37-3.42 (m, 1H), 3.50-3.5
5 (m, 1H), 3.79 (s, 6H), 3.98-4.02 (m, 6H), 4.12-
4.38 (m, 6H), 4.56 (dd, 2H, J = 4.8, 7.2Hz), 5.16-
5.20 (m, 1H), 5.28-5.35 (m, 1H), 6.21-6.26 (m, 1
H), 6.38-6.44 (m, 1H), 6.84 (d, 4H, J=7.6Hz), 7.
21-7.32 (m, 10H), 7.34-7.37 (m, 2H), 7.50-7.59
(m, 1H), 8.91-9.06 (m, 2H).
¹³ C-NMR (100 MHz, CDCl ₃ ): δ 11.72, 12.46, 14.13
, 19.59, 19.66, 19.75, 22.70, 26.08, 26.15, 29.
37, 29.46, 29.61, 29.67, 29.74, 30.36, 31.94, 3
3.94, 36.62, 38.95, 55.29, 59.89, 59.93, 62.39
, 62.45, 62.54, 62.59, 63.23, 67.62, 69.27, 73.
55, 74.09, 79.77, 79.82, 79.87, 79.92, 82.52, 8
4.29, 84.34, 84.46, 84.53, 85.36, 85.57, 87.34
, 108.14, 111.70, 111.82, 113.36, 116.17, 116.
36, 124.17, 127.34, 128.08, 128.14, 130.13, 13
4.94, 135.03, 135.07, 135.35, 135.49, 142.74,
143.97, 143.99, 150.23, 150.27, 150.38, 150.4
6, 152.89, 158.84, 163.40, 163.50, 166.14, 170
. 29, 170.34.
TOF/MS (ESI): _{calcd for C108H164N5O20PNa} [ _M
+ Na] ⁺ 1905.1605, found 1905.1636.

2-(N-methyl-3,4,5-tris(octadecyloxy)benzamido)acetic acid 5
Synthesis of '-O-((2-cyanoethoxy)(5'-O-(4,4'-dimethoxytrityl)thymidin-3'-yl)phosphoryl)thymidin-3'-yl [Example 5-9]
(1) 2-(N-methyl-3,4,5-tris(octadecyloxy)benzamide)
Synthesis of 5′-O-(4,4′-dimethoxytrityl)thymidin-3′-yl acetate

2-(N-methyl-3,4,5-tris(octadecyloxy)benzamido)acetic acid (
2.10 g, 2.10 mmol), 5′-O-(4,4′-dimethoxytrityl)thymidine (1.37 g, 2.52 mmol), 1-methylimidazole (0.86 g, 10.5
COMU (1.80 g, 4.20 mmol) in a suspension of COMU (1.80 g, 4.20 mmol) in THF (32 mL)
was added, and the mixture was stirred at room temperature for 3.5 hours. Furthermore, COMU (0.27 g, 0.63 mm
ol) was added, and the mixture was stirred at room temperature for 20 hours. Acetonitrile (158 mL) was added to the reaction solution.
was added dropwise to filter the precipitated solid. The solid was washed with an acetonitrile-THF mixed solvent and then dried under reduced pressure at 50° C. to obtain a crude product (2.96 g). The crude product was purified by column chromatography (neutral silica gel, developing solvent: dichloromethane-THF) to give the title compound (2.51 g, 78%) as a white solid.
¹ H-NMR (400 MHz, CDCl ₃ ): δ0.88 (t, 9H, J=6.8 Hz)
, 1.15-1.55 (m, 93H), 1.65-1.90 (m, 6H), 2.35-2
. 60 (m, 2H), 3.08 (s, 3H), 3.45-3.55 (m, 2H), 3.7
9 (s, 6H), 3.85-4.00 (m, 6H), 4.10-4.40 (m, 3H),
5.50-5.60 (m, 1H), 6.40-6.50 (m, 1H), 6.65 (s, 2
H), 6.80-6.90 (m, 4H), 7.20-7.40 (m, 9H), 7.62 (
s, 1H), 8.36 (s, 1H).
¹³ C-NMR (100 MHz, CDCl ₃ ): δ 11.64, 14.07, 22.71
, 26.17, 29.38, 29.48, 29.65, 29.70, 29.75, 30.
41, 31.97, 38.01, 55.30, 63.79, 69.54, 73.58, 7
6.72, 84.09, 84.54, 87.42, 106.36, 111.64, 113
. 49, 127.31, 128.09, 128.26, 129.90, 130.15, 1
35.30, 135.38, 140.18, 144.29, 150.20, 153.29
, 159.01, 163.21, 168.70, 172.20.
TOF/MS ( _ESI ): _{calcd for C95H149N3O12Na} [M+N
a] ⁺ 1547.1039, found 1547.1042.

[Example 5-10]
(2) 2-(N-methyl-3,4,5-tris(octadecyloxy)benzamide)
Synthesis of thymidin-3′-yl acetate

2-(N-methyl-3,4,5-tris(octadecyloxy)benzamido)acetic acid 5′-O-(4,4′-dimethoxytrityl)thymidine-3 according to the method of Example 2-2
'-yl (2.36 g, 1.55 mmol) gave the title compound (1.86 g, 98%) as a white solid.
¹ H-NMR (400 MHz, CDCl ₃ ): δ0.88 (t, 9H, J=6.8 Hz)
, 1.05-1.55 (m, 90H), 1.65-1.85 (m, 6H), 1.92 (s
, 3H), 2.30-2.60 (m, 3H), 3.09 (s, 3H), 3.85-4.0
5 (m, 8H), 4.05-4.35 (m, 3H), 5.45 (s, 1H), 6.05-
6.30 (brs, 1H), 6.64 (s, 2H), 7.44 (s, 1H), 8.28 (
s, 1H).
¹³ C-NMR (100 MHz, CDCl ₃ ): δ12.50, 14.08, 22.71
, 26.17, 29.38, 29.47, 29.51, 29.65, 29.70, 29.
75, 30.41, 31.97, 37.20, 39.06, 62.67, 69.59, 7
3.60, 75.74, 85.09, 86.62, 106.33, 111.45, 129
. 86, 136.38, 140.23, 150.26, 153.33, 163.22, 1
68.96, 172.29.
TOF/MS ( _ESI ): _{calcd for C74H131N3O10Na} [M+N
a] ⁺ 1244.9732, found 1244.9786.

[Example 5-11]
(3) 2-(N-methyl-3,4,5-tris(octadecyloxy)benzamide)
Synthesis of 5′-O-((2-cyanoethoxy)(5′-O-(4,4′-dimethoxytrityl)thymidin-3′-yl)phosphoryl)thymidin-3′-yl acetate

2-(N-Methyl-3,4,5-tris(octadecyloxy)benzamido)thymidin-3′-yl acetate (1.71 g, 1.40 mmol) and DMTr-dT- CE-phosphoramidite (2.09 g, 2.80 mmol) gave the title compound (2.54 g, 96%) as a white solid.
¹ H-NMR (400 MHz, CDCl ₃ ): δ0.88 (t, 9H, J=6.6 Hz)
, 1.05-1.55 (m, 93H), 1.65-1.85 (m, 6H), 1.91 (s
, 3H), 2.30-2.55 (m, 3H), 2.60-2.80 (m, 3H), 3.0
8 (s, 3H), 3.35-3.45 (m, 1H), 3.50-3.60 (m, 1H),
3.78 (s, 3H), 3.79 (s, 3H), 3.90-4.05 (m, 6H), 4.
10-4.45 (m, 8H), 5.10-5.20 (m, 1H), 5.30-5.45 (
m, 1H), 6.10-6.30 (brs, 1H), 6.35-6.45 (m, 1H),
6.64 (s, 2H), 6.75-6.90 (m, 4H), 7.15-7.40 (m, 1
0H), 7.49 (d, 1H, J=6.4Hz), 8.50-8.75 (m, 2H).
¹³ C-NMR (100 MHz, CDCl ₃ ): δ 11.76, 12.41, 12.46
, 14.07, 19.63, 19.70, 19.78, 19.85, 22.70, 26.
20, 29.37, 29.50, 29.52, 29.70, 29.75, 29.79, 3
0.43, 31.96, 36.86, 38.50, 39.03, 55.30, 55.34
, 61.99, 62.10, 62.50, 62.55, 62.63, 63.33, 67.
69, 69.61, 69.64, 73.60, 74.61, 79.78, 81.51, 8
2.43, 84.41, 84.52, 84.57, 85.65, 85.88, 86.36
, 87.47, 106.43, 111.40, 111.78, 111.83, 111.8
9, 113.31, 113.50, 116.07, 116.42, 127.09, 127
. 38, 127.86, 128.11, 128.27, 129.21, 130.20, 1
35.07, 135.18, 135.45, 135.54, 136.37, 139.66
, 140.30, 144.12, 147.51, 150.16, 150.21, 150.
26, 150.38, 153.30, 158.81, 159.03, 163.33, 16
8.90, 172.25, 172.43.
TOF/MS (ESI): _{calcd for C108H165N6O19PNa} [ _M
+ Na] ⁺ 1904.1765, found 1904.1780.

3-(3,4,5-tris(octadecyloxy)benzamido)propionic acid 5′-
Synthesis of O-((2-cyanoethoxy)(5′-O-(4,4′-dimethoxytrityl)thymidin-3′-yl)phosphoryl)thymidin-3′-yl [Example 5-12]
(1) Synthesis of 5′-O-(4,4′-dimethoxytrityl)thymidin-3′-yl 3-(3,4,5-tris(octadecyloxy)benzamido)propionate

3-(3,4,5-tris(octadecyloxy)benzamido)propionic acid (2.
20 g, 2.20 mmol), 5′-O-(4,4′-dimethoxytrityl)thymidine (
1.44 g, 2.64 mmol), 1-methylimidazole (0.90 g, 11.00 m
mol) in THF (44 mL) was added with COMU (1.88 g, 4.40 mmol) and stirred at room temperature for 3.5 hours. Furthermore, 1-methylimidazole (0.36 g, 4
. 4 mmol), COMU (0.94 g, 2.20 mmol) was added, and then 68 at room temperature.
Stirred for an hour. Acetonitrile (220 mL) was added dropwise to the reaction solution, and the precipitated solid was filtered. After washing the solid with acetonitrile-THF mixed solvent and methanol,
and dried under reduced pressure to obtain a crude product (3.07 g). The crude product was purified by column chromatography (neutral silica gel, developing solvent: dichloromethane-THF) to give the title compound (2.01 g, 60%) as a white solid.
¹ H-NMR (400 MHz, CDCl ₃ ): δ0.88 (t, 9H, J=6.8 Hz)
, 1.15-1.55 (m, 93H), 1.60-1.85 (m, 6H), 2.40-2
. 55 (m, 2H), 2.65-2.75 (m, 2H), 3.40-3.55 (m, 2H
), 3.65-3.75 (m, 2H), 3.78 (s, 6H), 3.95-4.05 (m
, 6H), 4.14 (d, 1H, J = 1.6Hz), 5.47 (d, 1H, J = 4.8H
z), 6.42 (t, 1H, J = 6.2Hz), 6.59 (t, 1H, J = 6.0Hz)
, 6.80-6.90 (m, 4H), 6.94 (s, 2H), 7.20-7.45 (m,
9H), 7.60 (s, 1H), 8.43 (s, 1H).
¹³ C-NMR (100 MHz, CDCl ₃ ): δ 11.62, 14.14, 22.71
, 26.11, 29.38, 29.45, 29.61, 29.68, 29.74, 30.
33, 31.94, 34.05, 35.35, 37.92, 55.24, 63.65, 6
9.36, 73.51, 75.85, 83.94, 84.34, 87.24, 105.6
7, 111.65, 113.15, 113.32, 127.26, 127.76, 127
. 85, 128.06, 128.10, 129.12, 130.04, 130.08, 1
35.03, 135.24, 135.35, 141.26, 144.12, 150.23
, 153.10, 158.78, 163.39, 167.36, 172.25.
TOF/MS ( _ESI ): _{calcd for C95H149N3O12Na} [M+N
a] ⁺ 1547.1039, found 1547.1014.

[Example 5-13]
(2) Synthesis of thymidin-3′-yl 3-(3,4,5-tris(octadecyloxy)benzamido)propionate

3-(3,4,5-Tris(octadecyloxy)benzamido)propionate 5′-O-(4,4′-dimethoxytrityl)thymidine-3′- according to the method of Example 2-2
yl (1.91 g, 1.25 mmol) gave the title compound (1.51 g, 99%) as a white solid.
¹ H-NMR (400 MHz, CDCl ₃ ): δ0.88 (t, 9H, J=6.8 Hz)
, 1.15-1.55 (m, 90H), 1.65-1.85 (m, 6H), 1.91 (s
, 3H), 2.30-2.55 (m, 2H), 2.55-2.75 (m, 3H), 3.6
5-3.75 (m, 2H), 3.85-3.95 (m, 2H), 3.95-4.05 (m
, 6H), 4.05-4.10 (m, 1H), 5.35-5.40 (m, 1H), 6.1
6 (dd, 1H, J = 6.0, 8.4Hz), 6.55-6.65 (m, 1H), 6.9
5 (s, 2H), 7.43 (s, 1H), 8.25-8.55 (m, 1H).
¹³ C-NMR (100 MHz, CDCl ₃ ): δ12.51, 14.09, 22.71
, 26.13, 26.16, 29.38, 29.47, 29.62, 29.69, 29.
75, 29.99, 30.38, 31.96, 34.26, 35.57, 37.12, 6
2.57, 69.57, 73.59, 75.09, 85.06, 86.65, 106.0
4, 111.41, 129.08, 136.51, 141.64, 150.33, 153
. 20, 163.40, 167.46, 172.39.
TOF/MS ( _ESI ): _{calcd for C74H131N3O10Na} [M+N
a] ⁺ 1244.9732, found 1244.9789.

[Example 5-14]
(3) 3-(3,4,5-tris(octadecyloxy)benzamido)propionic acid 5′-O-((2-cyanoethoxy)(5′-O-(4,4′-dimethoxytrityl)thymidine- Synthesis of 3′-yl)phosphoryl)thymidin-3′-yl

3-(3,4,5-Tris(octadecyloxy)benzamido)thymidin-3′-ylpropionate (1.41 g, 1.15 mmol) and DM according to the method of Example 3-4.
Tr-dT-CE-phosphoramidite (1.71 g, 2.30 mmol) gave the title compound (2.07 g, 95%) as a white solid.
¹ H-NMR (400 MHz, CDCl ₃ ): δ0.88 (t, 9H, J=6.8 Hz)
, 1.15-1.55 (m, 93H), 1.65-1.85 (m, 7H), 1.90 (d
, 3H, J=1.2Hz), 2.25-2.55(m, 3H), 2.55-2.80(m
, 5H), 3.30-3.45 (m, 1H), 3.50-3.60 (m, 1H), 3.6
0-3.75 (m, 2H), 3.78 (s, 3H), 3.79 (s, 3H), 3.90-
4.05 (m, 6H), 4.05-4.40 (m, 6H), 5.10-5.25 (m, 1
H), 5.30-5.40 (m, 1H), 6.15-6.25 (m, 1H), 6.40-
6.50 (m, 1H), 6.75-6.90 (m, 4H), 6.99 (d, 2H, J = 3
. 6Hz), 7.20-7.40 (m, 9H), 7.50-7.60 (m, 1H), 8.
95-9.05 (m, 1H), 9.05-9.15 (m, 1H).
¹³ C-NMR (100 MHz, CDCl ₃ ): δ 11.72, 12.49, 14.14
, 19.59, 19.67, 19.75, 22.71, 26.11, 26.14, 29.
38, 29.47, 29.63, 29.69, 29.75, 29.78, 30.35, 3
1.94, 34.03, 35.53, 35.58, 36.64, 38.95, 55.29
, 62.36, 62.41, 62.52, 62.57, 63.24, 67.60, 69.
38, 73.53, 73.84, 79.90, 82.35, 82.44, 84.25, 8
4.32, 84.39, 85.48, 85.67, 87.35, 105.86, 111.
64, 111.74, 111.88, 113.14, 113.35, 116.25, 11
6.44, 127.34, 127.76, 127.85, 128.09, 129.03,
129.13, 130.13, 134.91, 134.95, 135.00, 135.0
7, 135.48, 135.61, 141.22, 143.93, 143.97, 150
. 21, 150.26, 150.46, 150.49, 153.03, 158.83, 1
63.48, 163.53, 167.47, 167.51, 172.19.
TOF/MS (ESI): _{calcd for C108H165N6O19PNa} [ _M
+ Na] ⁺ 1904.1765, found 1904.1807.

6.2-((3,4,5-tris(octadecyloxy)benzoyl)oxy)acetic acid (
4-(((6-(N ⁶ -benzoyladenin-9-yl)-4-tritylmorpholine-2
Synthesis of -yl)methoxy)(dimethylamino)phosphoryl)-6-(thymin-1-yl)morpholin-2-yl)methyl [Example 6-1]
(1) Synthesis of (6-(thymin-1-yl)-4-tritylmorpholin-2-yl)methyl 2-((3,4,5-tris(octadecyloxy)benzoyl)oxy)acetate

According to the method of Example 2-1, 2-((3,4,5-tris(octadecyloxy)benzoyl)oxy)acetic acid (1.97 g, 2.00 mmol) and 6-(hydroxymethyl)
-4-tritylmorpholin-2-yl)-5-methylpyrimidine-2,4-(1H,3H
)-dione (1.16 g, 2.40 mmol) to title compound (2.85 g, 98%)
got
¹ H-NMR (400 MHz, CDCl ₃ ): δ0.88 (t, 9H, J=6.8 Hz)
, 1.11-1.53 (m, 91H), 1.68-1.83 (m, 6H), 1.84 (s
, 3H), 3.13 (d, 1H, J=12.0 Hz), 3.37 (d, 1H, J=11.
2Hz), 3.98-4.04 (m, 6H), 4.17 (d, 2H, J = 5.2Hz),
4.37-4.41 (m, 1H), 4.78 (s, 2H), 6.13 (dd, 1H, J=
2.0, 9.6Hz), 7.00 (s, 1H), 7.15-7.26 (m, 3H), 7.
26-7.38 (m, 8H), 7.34-7.58 (m, 6H), 8.34 (s, 1H)
.
¹³ C-NMR (100 MHz, CDCl ₃ ): δ12.45, 14.13, 22.70
, 26.07, 26.11, 29.32, 29.38, 29.42, 29.59, 29.
66, 29.68, 29.73, 30.35, 31.94, 49.01, 51.85, 6
0.89, 65.08, 69.19, 73.55, 74.58, 80.50, 108.3
1, 110.68, 123.33, 126.64, 127.97, 129.13, 135
. 33, 142.95, 149.63, 152.91, 163.23, 165.75, 1
67.64.
TOF/MS ( _ESI ): _calcd for _{C92H143N3O10Na} [ _M +N
a] ⁺ 1473.0671, found 1473.0629.

[Example 6-2]
(2) Synthesis of 2-((3,4,5-tris(octadecyloxy)benzoyl)oxy)acetic acid (6-(thymin-1-yl)morpholin-2-yl)methyl trifluoroacetate

(6-(thymin-1-yl)-4-tritylmorpholin-2-yl)methyl 2-((3,4,5-tris(octadecyloxy)benzoyl)oxy)acetate according to the method of Example 2-2 (2.76 g, 1.90 mmol) to the title compound (2.44 g, 97%)
got
¹ H-NMR (400 MHz, CDCl ₃ ): δ0.88 (t, 9H, J=6.8 Hz)
, 1.10-1.39 (m, 84H), 1.42-1.50 (m, 6H), 1.70-1
. 83 (m, 6H), 1.87 (s, 3H), 2.95-3.16 (m, 2H), 3.4
2 (d, 1H, J=11.2Hz), 3.62-3.75 (m, 1H), 3.95-4.
05 (m, 6H), 4.31 (d, 1H, J = 8.4Hz), 4.46 (d, 2H, J =
10.0 Hz), 4.86 (s, 2H), 6.17 (d, 1H, J=9.6 Hz), 7.
25 (s, 1H), 7.30 (s, 2H), 10.15-10.75 (brs, 1H).
¹³ C-NMR (100 MHz, CDCl ₃ ): δ12.48, 14.13, 22.71
, 26.09, 26.15, 29.35, 29.39, 29.47, 29.63, 29.
69, 29.75, 29.78, 30.38, 31.94, 43.24, 44.99, 6
0.87, 63.56, 69.21, 72.38, 73.57, 108.30, 112.
18, 114.99, 117.90, 123.12, 134.33, 143.10, 15
0.40, 152.99, 162.55, 162.90, 163.78, 166.02,
167.45.
TOF/MS ₍ ESI): _calcd for _{C73H129N3O10 - CF3CO}
OH [MH] ^- 1320.9528, found 1320.9517.

[Example 6-3]
(3) 2-((3,4,5-tris(octadecyloxy)benzoyl)oxy)acetic acid (4-(((6-(N ⁶ -benzoyladenin-9-yl)-4-tritylmorpholine-)
Synthesis of 2-yl)methoxy)(dimethylamino)phosphoryl)-6-(thymin-1-yl)morpholin-2-yl)methyl

2-((3,4,5-tris(octadecyloxy)benzoyl)oxy)acetic acid (6-
(thymin-1-yl)morpholin-2-yl)methyltrifluoroacetate (2.32 g,
1.76 mmol), N,N-diisopropylethylamine (4.16 g, 32.16 m
mol) of 1,3-dimethyl-2-imidazolidinone (16 mL)-THF solution (18 mL
L) to (6-(N ⁶ -benzoyladenin-9-yl)-4-tritylmorpholine-2-
yl)methyldimethylphosphoroamide chloride (3.81 g, 5.27 mmol) was added and stirred at room temperature for 2.5 hours. The slurry obtained by adding methanol (93 mL) to the reaction solution was concentrated under reduced pressure. A slurry obtained by adding methanol to the concentrated residue was stirred at room temperature for 30 minutes and then filtered. After washing the solid with methanol, it was dried under reduced pressure at 50° C. to obtain a crude product (3.23 g). The title compound (
1.96 g, 59%) as a white solid.
¹ H-NMR (400 MHz, CDCl ₃ ): δ0.88 (t, 9H, J=6.8 Hz)
, 1.10-1.53 (m, 90H), 1.55-1.97 (m, 12H), 2.46 (
d, 3H, J = 10.4 Hz), 2.63 (d, 3H, J = 9.6 Hz), 2.46-2
. 63 (m, 1H), 3.12-3.60 (m, 4H), 3.75-4.07 (m, 10
H), 4.15-4.50 (m, 2H), 4.78-4.91 (m, 2H), 5.41-
5.53 (m, 1H), 6.42 (d, 1H, J=10.4Hz), 7.12-7.20
(m, 4H), 7.20-7.32 (m, 9H), 7.40-7.58 (m, 8H), 8
. 05 (d, 2H, J=7.6Hz), 8.07-8.10 (m, 1H), 8.68-8
. 80 (m, 1H), 9.50-9.76 (m, 2H).
¹³ C-NMR (100 MHz, CDCl ₃ ): δ12.44, 12.53, 14.14
, 22.70, 26.08, 26.12, 29.32, 29.38, 29.43, 29.
60, 29.68, 29.73, 30.35, 31.94, 36.47, 36.51, 3
6.63, 36.67, 44.56, 44.84, 47.02, 48.64, 48.78
, 52.81, 53.10, 60.93, 64.23, 65.34, 65.69, 69.
19, 73.56, 74.90, 74.97, 75.34, 79.53, 79.61, 7
9.97, 80.79, 108.27, 110.92, 111.25, 122.82, 1
23.26, 126.65, 128.00, 128.22, 128.29, 128.57
, 128.63, 129.16, 132.47, 132.57, 133.49, 133.
92, 134, 74, 134.80, 140.75, 140.78, 142.97, 14
3.00, 149.31, 149.68, 149.82, 150.12, 150.89,
151.40, 152.65, 152.77, 152.92, 152.94, 163.6
2, 165.02, 165.23, 165.82, 165.84, 167.56, 167
. 63.
TOF/MS ( _ESI ): calcd _{for C111H165N10O14PNa [}
M+Na] ⁺ 1916.2142, found 1916.2216.

2-((3,4,5-tris(octadecyloxy)benzoyl)oxy)acetic acid (4-
(((6-(N ⁴ -benzoylcytosin-1-yl)-4-tritylmorpholin-2-yl)methoxy)(dimethylamino)phosphoryl)-6-(N ² -isobutyrylguanine-
Synthesis of 9-yl)morpholin-2-yl)methyl [Example 6-4]
(1) (6-(N ² -isobutyrylguanin-9-yl)-4-tritylmorpholin-2-yl)methyl 2-((3,4,5-tris(octadecyloxy)benzoyl)oxy)acetate Synthesis of

2-((3,4,5-Tris(octadecyloxy)benzoyl)oxy)acetic acid (1.97 g, 2.00 mmol) and N-(9-(6-(hydroxymethyl )-4-Tritylmorpholin-2-yl))-6-oxo-6,9-dihydro-1H-purin-2-yl)isobutyramide (1.51 g, 2.60 mmol) to the title compound (3.00 g) , 97%).
¹ H-NMR (400 MHz, CDCl ₃ ): δ0.88 (t, 9H, J=6.8 Hz)
, 1.10-1.60 (m, 97H), 1.70-1.90 (m, 7H), 2.64 (s
ept, 1H), 3.18 (d, 1H, J = 12.0 Hz), 3.45 (d, 1H, J =
11.6 Hz), 3.95-4.15 (m, 6H), 4.13-4.25 (m, 2H),
4.37-4.46 (m, 1H), 4.72 (d, 1H, J=16.0Hz), 4.84
(d, 1H, J = 16.0Hz), 5.89 (dd, 1H, J = 2.2, 9.8Hz),
7.06-7.37 (m, 12H), 7.39-7.55 (m, 5H), 7.61 (s,
1H), 8.18 (s, 1H), 11.92 (s, 1H).
¹³ C-NMR (100 MHz, CDCl ₃ ): δ 14.14, 18.93, 19.03
, 22.71, 26.07, 26.12, 29.33, 29.37, 29.42, 29.
60, 29.67, 29.74, 30.35, 31.94, 36.64, 49.04, 5
2.24, 60.90, 65.03, 69.25, 73.58, 74.22, 80.36
, 108.38, 121.30, 123.27, 126.69, 127.96, 129.
18, 136.12, 143.08, 147.32, 147.42, 152.95, 15
5.36, 165.98, 167.61, 177.85.
TOF/MS ( _ESI ): _{calcd for C96H148N6O10Na} [M+N
a] ⁺ 1568.1155, found 1568.1176.

[Example 6-5]
(2) 2-((3,4,5-tris(octadecyloxy)benzoyl)oxy)acetic acid (6-(N ² -isobutyrylguanin-9-yl)morpholin-2-yl)methyltrifluoroacetate Synthesis of

2-((3,4,5-tris(octadecyloxy)benzoyl)oxy)acetic acid (6-(N ² -isobutyrylguanin-9-yl)-4-tritylmorpholine according to the method of Example 2-2 -2-yl)methyl (2.78 g, 1.80 mmol) to the title compound (
2.45 g, 96%).
¹ H-NMR (400 MHz, CDCl ₃ ): δ0.87 (t, 9H, J=6.8 Hz)
, 1.00-1.53 (m, 96H), 1.69-1.88 (m, 6H), 2.72-2
. 88 (m, 1H), 3.19-3.38 (m, 1H), 3.44-3.63 (m, 1H
), 3.75-4.13 (m, 8H), 4.25-4.63 (m, 3H), 4.69-4
. 94 (m, 2H), 6.06-6.31 (m, 1H), 7.20-7.30 (m, 3H
), 7.87-8.06 (m, 1H), 10.19-10.63 (brs, 1H), 12
. 19-12.63 (brs, 1H).
¹³ C-NMR (100 MHz, CDCl ₃ ): δ 14.12, 18.92, 22.70
, 26.09, 26.16, 29.35, 29.38, 29.47, 29.63, 29.
68, 29.75, 29.79, 30.04, 30.38, 31.94, 36.17, 4
2.86, 60.73, 63.62, 69.21, 71.44, 73.57, 108.2
9, 115.13, 118.04, 123.10, 143.09, 148.28, 152
. 95, 156.00, 165.93, 167.50, 180.39.
TOF/MS ₍ ESI): calcd _{for C77H134N6O10 - CF3CO}
OH [M-H] ^- 1416.0012, found 1415.9960.

[Example 6-6]
(3) 2-((3,4,5-tris(octadecyloxy)benzoyl)oxy)acetic acid (4-(((6-(N ⁴ -benzoylcytosin-1-yl)-4-tritylmorpholine-)
Synthesis of 2-yl)methoxy)(dimethylamino)phosphoryl)-6-(N ² -isobutyrylguanin-9-yl)morpholin-2-yl)methyl

2-((3,4,5-tris(octadecyloxy)benzoyl)oxy)acetic acid (6-(N ² -isobutyrylguanin-9-yl)morpholine-
2-yl)methyl trifluoroacetate (2.27 g, 1.60 mmol) and (6-(N ⁴
-benzoylcytosin-1-yl)-4-tritylmorpholin-2-yl)methyldimethylphosphoroamide chloride (3.35 g, 4.80 mmol) to the title compound (2).
. 87 g, 91%).
¹ H-NMR (400 MHz, CDCl ₃ ): δ0.88 (t, 9H, J=6.8 Hz)
, 1.00-1.56 (m, 96H), 1.69-1.88 (m, 9H), 2.38-2
. 97 (m, 8H), 3.06-3.69 (m, 5H), 3.75-4.56 (m, 12
H), 4.69-4.94 (m, 2H), 5.34-5.63 (m, 1H), 6.28-
6.50 (m, 1H), 7.13-8.00 (m, 23H), 8.69-8.94 (br
s, 1H), 9.31-9.63 (brs, 1H), 10.06-10.31 (brs,
1H), 12.13-12.31 (m, 1H).
¹³ C-NMR (100 MHz, CDCl ₃ ): δ 14.13, 19.01, 19.04
, 19.17, 19.22, 22.70, 26.07, 26.12, 29.32, 29.
37, 29.43, 29.60, 29.67, 29.73, 29.98, 30.35, 3
1.94, 36.17, 36.27, 36.64, 36.68, 36.74, 36.78
, 45.00, 45.08, 46.96, 47.43, 48.25, 48.76, 52.
48, 60.82, 60.90, 64.27, 65.84, 69.20, 69.22, 7
3.57, 74.31, 74.51, 75.41, 79.30, 80.47, 81.36
, 81.59, 108.31, 121.25, 122.00, 123.26, 123.3
3, 126.67, 126.73, 127.73, 127.99, 128.80, 128
. 98, 132.71, 133.07, 133.27, 136.19, 137.10, 1
42.90, 143.01, 147.88, 148.00, 148.09, 152.89
, 152.93, 155.79, 155.90, 165.79, 165.91, 167.
56, 167.58, 178.67, 179.37.
TOF/MS ( _ESI ): calcd _{for C114H170N11O15PNa [}
M+Na] ⁺ 1987.2513, found 1987.2521.

7. Confirmation of detagging [Example 7-1]
5′-O-((2-cyanoethoxy)(5′-O-((2-cyanoethoxy)(5′-
Synthesis ^of O-(4,4′-dimethoxytrityl)thymidin-3′-yl)phosphoryl)-N 2 -isobutyryldeoxyguanosin-3′-yl)phosphoryl)thymidine

2-((3,4,5-tris(octadecyloxy)benzoyl)oxy)acetic acid 5′-
O-((2-cyanoethoxy)(5'-O-((2-cyanoethoxy)(5'-O-(4
, 4′-dimethoxytrityl)thymidin-3′-yl)phosphoryl)-N ² -isobutyryldeoxyguanosin-3′-yl)phosphoryl)thymidin-3′-yl (233 mg,
0.10 mmol) in THF (9 mL)-2-propanol (1 mL) was cooled to 0°C. Under cooling, 4 mol / L-lithium borohydride-THF solution (0.125 m
L, 0.50 mmol) was added, and the mixture was stirred at the same temperature for 30 minutes. After adding an aqueous solution of ammonium chloride to the reaction solution, extraction was performed three times with dichloromethane. The organic layer was washed with brine, dried over anhydrous sodium sulfate, filtered to remove sodium sulfate, and the filtrate was concentrated. Remove the solid precipitated by suspending the concentrated residue in acetonitrile by filtration,
The filtrate was concentrated. Dimethyl sulfoxide of concentrated residue - methanol mixed solution (1: 1, 5m
A portion (2.5 mL) of L) was purified by preparative chromatography in 5 portions to give the title compound (64 mg, 95%) as a white solid.

Preparative chromatography conditions: column Kanto Kagaku Mightysil RP-18GP (5 μm, 2
0 Φ × 250 mm), flow rate 5 mL / min, mobile phase acetonitrile-water (gradient 0
-30 min (proportion of acetonitrile 50→60%).
¹ H-NMR (600 MHz, DMSO-d ₆ ): δ 1.12 (d, 6H, J = 6.0H
z), 1.42 (d, 3H, J = 7.2Hz), 1.74 (d, 3H, J = 2.8Hz)
, 2.07 (s, 1H), 2.30-2.41 (m, 3H), 2.60-2.90 (m,
6H), 3.18-3.29 (m, 2H), 3.73 (d, 6H, J=4.2Hz), 4
. 00-4.43 (m, 12H), 4.97 (s, 1H), 5.10 (s, 1H), 5.
51-5.52 (m, 1H), 6.15-6.25 (m, 3H), 6.88 (d, 4H,
J=9.0 Hz), 7.20-7.50 (m, 11H), 8.17 (q, 1H, J=1.
8 Hz), 11.33-12.07 (m, 3H).
¹³ C-NMR (151 MHz, DMSO-d ₆ ): δ 10.56, 10.88, 17.
66, 33.65, 35.78, 52.00, 53.86, 56.34, 61.48, 6
1.80, 65.49, 66.58, 68.73, 76.58, 77.11, 81.63
, 82.11, 82.52, 82.87, 84.98, 90.73, 108.72, 11
2.08, 116.98, 119.14, 125.69, 126.46, 126.77,
128.53, 133.81, 134.00, 134.26, 134.70, 136.1
5, 143.26, 147.04, 147.32, 149.14, 153.53, 156
. 99, 162.50, 168.14, 178.89.
TOF/MS ₍ ESI): _{calcd for C61H69N11O21P2Na [}
M+Na] ⁺ 1376.4, found 1376.6.

[Example 7-2]
5'-O-((2-cyanoethoxy)(5'-O-((2-cyanoethoxy)(5'-
O-(4,4'-dimethoxytrityl)-N ² -isobutyryldeoxyguanosine-3'
-yl)phosphoryl)thymidin-3′-yl)phosphoryl)-N ⁶ -benzoyldeoxyadenosine

2-((3,4,5-tris(octadecyloxy)benzoyl)oxy)acetic acid 5′-O-((2-cyanoethoxy)(5′-O-((2 -cyanoethoxy ⁾ (5′-O-(4,4′-dimethoxytrityl)-N 2 -isobutyryldeoxyguanosin-3′-yl)phosphoryl)thymidin-3′-yl)phosphoryl)-N ⁶
-benzoyldeoxyadenosin-3′-yl (244 mg, 0.10 mmol) gave the title compound (55 mg, 75%) as a yellow solid.
¹ H-NMR (600 MHz, DMSO-d ₆ ): δ 1.17-1.30 (m, 7H),
1.74-1.79 (m, 3H), 1.97-2.30 (m, 2H), 2.45-2.5
1 (m, 2H), 2.68-2.76 (m, 1H), 2.79-2.83 (m, 1H),
2.96 (dt, 4H, J=6.4, 12.4Hz), 3.03-3.12 (m, 1H)
, 3.22(t, 1H, J=11.3 Hz), 3.77(s, 6H), 3.93(s, 1
H), 4.07-4.38 (m, 11H), 5.07-5.45 (m, 5H), 6.05
-6.45 (m, 3H), 6.85 (t, 4H, J = 10.7 Hz), 7.24 (d, 5
H, J = 8.9 Hz), 7.29 (t, 2H, J = 7.6 Hz), 7.36 (d, 2H,
J = 7.6 Hz), 7.49-7.56 (m, 3H), 7.65 (t, 1H, J = 7.2
Hz), 7.90 (dt, 1H, J = 3.0, 8.1 Hz), 8.07 (d, 2H, J =
7.6Hz), 8.13-8.16 (m, 1H), 10.89-12.13 (m, 3H)
.
¹³ C-NMR (151 MHz, DMSO-d ₆ ): δ 12.52, 14.58, 19.
33, 35.33, 36.13, 37.02, 55.48, 60.34, 62.73, 6
3.24, 64.06, 67.31, 68.33, 70.66, 77.30, 79.12
, 82.64, 83.37, 84.09, 84.74, 86.25, 113.58, 11
8.69, 121.00, 123.54, 127.24, 128.19, 128.72,
128.94, 130.23, 132.68, 133.50, 134.55, 135.7
7, 136.46, 137.81, 139.68, 145.08, 148.56, 149
. 12, 150.82, 155.31, 158.56, 161.08, 164.11, 1
65.03, 180.61.
TOF/MS _{( ESI} ): _{calcd for C68H72N14O20P2Na} [
M+Na] ⁺ 1489.4, found 1489.4.

[Example 7-3]
5′-O-((2-cyanoethoxy)(5′-O-((2-cyanoethoxy)(5′-
Synthesis of O-(4,4′-dimethoxytrityl)thymidin-3′-yl)phosphoryl)-N ² -isobutyryldeoxyguanosin-3′-yl)phosphoryl)-N ⁴ -benzoyldeoxycytidine

2-((3,4,5-tris(octadecyloxy)benzoyl)oxy)acetic acid 5′-O-((2-cyanoethoxy)(5′-O-((2 -cyanoethoxy)(5′-O-(4,4′-dimethoxytrityl)thymidin-3′-yl)phosphoryl)-N ² -isobutyryldeoxyguanosin-3′-yl)phosphoryl)-N ⁴
-benzoyldeoxycytidin-3′-yl (243 mg, 0.10 mmol) gave the title compound (57 mg, 79%) as a white solid.
¹ H-NMR (600 MHz, DMSO-d ₆ ): δ 1.09-1.12 (m, 6H),
1.42 (s, 3H), 1.75-2.10 (m, 4H), 2.61-2.95 (m, 7
H), 3.15-3.25 (m, 3H), 3.70-3.77 (m, 7H), 4.08-
4.36 (m, 11H), 5.09 (d, 2H, J=41.9Hz), 5.32 (d, 2
H, J = 44.0 Hz), 6.18-6.29 (m, 3H), 6.85-6.88 (m,
4H), 7.20-7.22 (m, 5H), 7.29 (q, 2H, J=6.9Hz), 7
. 34 (d, 2H, J = 7.6Hz), 7.41-7.53 (m, 4H), 7.86 (d
, 2H, J=7.6 Hz), 8.19 (dt, 1H, J=4.4, 8.2 Hz), 8.8
4 (q, 1H, J=6.4Hz), 11.34-12.05 (m, 3H).
¹³ C-NMR (151 MHz, DMSO-d ₆ ): δ 12.17, 19.35, 27.
86, 35.35, 55.56, 57.07, 63.19, 63.61, 67.29, 6
8.60, 70.86, 78.19, 78.93, 82.62, 83.40, 83.86
, 84.14, 86.67, 110.47, 113.78, 118.59, 118.88
, 120.91, 127.38, 128.02, 128.69, 130.24, 131.
86, 134.55, 135.51, 135.97, 137.92, 144.97, 14
8.76, 149.02, 150.84, 154.36, 155.25, 158.69,
164.09, 165.95, 166.18, 180.60.
TOF/MS ₍ ESI): _{calcd for C67H72N12O21P2Li [}
M+Li] ⁺ 1449.5, found 1449.6.

[Example 7-4]
5'-O-((2-cyanoethoxy)(5'-O-((2-cyanoethoxy)(5'-
O-(4,4'-dimethoxytrityl)thymidin-3'-yl)phosphoryl)thymidine-
Synthesis of 3′-yl)phosphoryl)-N ² -isobutyryldeoxyguanosine

2-((3,4,5-tris(octadecyloxy)benzoyl)oxy)acetic acid 5′-O-((2-cyanoethoxy)(5′-O-((2 -cyanoethoxy)(5′-O-(4,4′-dimethoxytrityl)thymidin-3′-yl)phosphoryl)thymidin-3′-yl)phosphoryl)-N ² -isobutyryldeoxyguanosine-3′- title compound (63 mg, 93%) from yl (232 mg, 0.10 mmol)
) as a white solid.
¹ H-NMR (600 MHz, DMSO-d ₆ ): δ 1.12 (d, 6H, J = 6.2H
z), 1.42 (d, 3H, J = 7.6 Hz), 1.73 (d, 3H, J = 2.7 Hz)
, 2.07 (s, 1H), 2.32-2.43 (m, 3H), 2.61-2.91 (m,
6H), 3.20-3.29 (m, 2H), 3.73 (d, 6H, J=3.8Hz), 4
. 04-4.43 (m, 12H), 4.97 (s, 1H), 5.10 (s, 1H), 5.
51-5.52 (m, 1H), 6.13-6.26 (m, 3H), 6.88 (d, 4H,
J = 8.9Hz), 7.23 (d, 5H, J = 7.6Hz), 7.30 (t, 2H, J =
6.2Hz), 7.36 (d, 2H, J = 8.2Hz), 7.43-7.47 (m, 2H
), 7.89-8.18 (m, 1H), 11.33-12.07 (m, 3H).
¹³ C-NMR (151 MHz, DMSO-d ₆ ): δ 10.46, 10.84, 17.
69, 17.83, 33.61, 35.32, 36.22, 53.86, 61.50, 6
1.98, 65.53, 66.44, 68.89, 75.88, 77.15, 80.92
, 81.78, 82.16, 82.47, 82.92, 83.51, 85.00, 108
. 75, 108.90, 112.08, 116.92, 119.11, 125.71, 1
26.48, 126.77, 128.55, 133.80, 134.02, 134.33
, 134.53, 136.24, 143.27, 146.96, 147.20, 149.
17, 153.63, 157.01, 162.41, 178.92.
TOF/MS ( _ESI ): _{calcd for C61H69N11O21P2Na [}
M+Na] ⁺ 1376.4, found 1376.6.

[Example 7-5]
5'-O-((2-cyanoethoxy)(5'-O-(4,4'-dimethoxytrityl)
-thymidin-3′-yl)phosphorothioyl)-thymidine

2-((3,4,5-tris(octadecyloxy)benzoyl)oxy)acetic acid 5′-
O-((2-cyanoethoxy)(5′-O-(4,4′-dimethoxytrityl)-thymidin-3′-yl)phosphorothioyl)-thymidin-3′-yl (189 mg, 0.10 m
mol) in THF (9 mL)-2-propanol (1 mL) was cooled to 0°C.
Under cooling, 4 mol/L-lithium borohydride-THF solution (0.125 mL, 0.5
0 mmol) was added, and the mixture was stirred at the same temperature for 30 minutes. After adding an aqueous solution of ammonium chloride to the reaction solution, extraction was performed three times with dichloromethane. The organic layer was washed with brine, dried over anhydrous sodium sulfate, filtered to remove sodium sulfate, and the filtrate was concentrated. The solid precipitated by suspending the concentrated residue in acetonitrile was removed by filtration, and the filtrate was concentrated. The concentrated residue was purified by column chromatography (silica gel, developing solvent: chloroform-methanol) to give the title compound (86 mg, 93%) as a white solid.
¹ H-NMR (600 MHz, CDCl ₃ ): δ 1.46 (d, 3H, J = 15.8 Hz
), 1.91 (d, 3H, J=8.2Hz), 2.21-2.79 (m, 6H), 3.3
9-3.49 (m, 2H), 3.79 (d, 6H, J=3.4Hz), 4.08-4.5
2 (m, 8H), 5.26-5.31 (m, 1H), 6.25-6.41 (m, 2H),
6.85 (dd, 4H, J=2.7, 8.9Hz), 7.22-7.38 (m, 9H),
7.58 (d, 1H, J=24.1Hz), 9.45-10.50 (m, 2H).
¹³ C-NMR (151 MHz, CDCl ₃ ): δ 11.90, 12.57, 19.43
, 38.92, 39.90, 55.30, 62.59, 63.36, 67.70, 71.
03, 80.38, 84.66, 85.46, 87.28, 111.32, 111.97
, 113.38, 116.54, 127.28, 128.00, 128.11, 130.
06, 135.00, 135.80, 144.09, 150.64, 150.92, 15
8.74, 163.98.
TOF/MS ( _ESI ) _: calcd for _{C44H48N5O13PSNa} [ _M
+ Na] ⁺ 940.3, found 940.2.

[Example 7-6]
5'-O-((2-cyanoethoxy)(5'-O-(4,4'-dimethoxytrityl)
-2′-fluorodeoxyuridin-3′-yl)phosphoryl)-2′-fluoro-N ²
- Synthesis of isobutyryldeoxyguanosine

2-((3,4,5-tris(octadecyloxy)benzoyl)oxy)acetic acid 5′-
O-((2-cyanoethoxy)(5'-O-(4,4'-dimethoxytrityl)-2'-
fluorodeoxyuridin-3′-yl)phosphoryl)-2′-fluoro-N ² -isobutyryldeoxyguanosin-3′-yl (0.397 g, 0.200 mmol) in THF
A mixed solution of (27 mL)-2-propanol (3 mL) was cooled to 0 to -10°C. Under cooling, 4 mol/L-lithium borohydride-THF solution (0.10 mL, 0.400 m
mol) was added, and the mixture was stirred at the same temperature for 20 minutes. A 10% aqueous ammonium chloride solution (4 mL) was added to the reaction mixture, and the mixture was separated, and the organic layer was washed with 10% brine. The organic layer was dried over anhydrous sodium sulfate, filtered to remove sodium sulfate, and the filtrate was concentrated. The solid precipitated by adding methanol to the concentrated residue was removed by filtration, and the filtrate was concentrated to dryness. The concentrated residue was purified by column chromatography (neutral silica gel, developing solvent: dichloromethane-methanol) to give the title compound (0.137 g, 67%) as a white solid.
¹ H-NMR (400 MHz, CDCl ₃ ): δ1.16-1.26 (m, 6H),2.
19 (s, 2H), 2.45-2.87 (m, 3H), 3.44-3.66 (m, 2H)
, 3.72-3.75 (m, 6H), 3.92-4.18 (m, 2H), 4.25-4.
50 (m, 4H), 4.89-5.55 (m, 6H), 5.85-6.00 (m, 2H)
，
6.80-6.82 (m, 4H), 7.15-7.30 (m, 7H), 7.33-7.3
7 (m, 2H), 7.71-7.82 (m, 2H), 10.05-10.42 (brs,
1H), 10.55-10.87 (brs, 1H), 12.36 (s, 1H).
¹³ C-NMR (100 MHz, CDCl ₃ ): δ 18.75, 18.85, 19.01
, 19.07, 19.38, 19.45, 36.01, 50.81, 55.26, 55.
28, 60.65, 62.85, 67.86, 80.26, 81.13, 87.13, 8
7.19, 87.53, 87.88, 90.59, 92.53, 102.62, 113.
33, 116.74, 121.28, 127.26, 128.05, 128.19, 13
0.22, 134.80, 134.91, 134.96, 135.07, 140.41,
144.01, 144.11, 148.00, 148.06, 148.50, 148.5
5, 150.44, 155.70, 158.76, 163.72, 180.33, 180
. 39.
TOF _/ MS (ESI): _{calcd for C47H49N8O14F2PNa [}
M+Na] ⁺ 1041.2972, found 1041.2975.

[Example 7-7]
N ⁴ -benzoyl-5′-O-((2-cyanoethoxy)(5′-O-(4,4′-dimethoxytrityl)-2′-O-methyluridin-3′-yl)phosphoryl)-2 '-O
- Synthesis of methylcytidine

N,N-diisopropylethylamine (0.065 g, 0.50 mmol) in THF (
13.5 mL) and 2-propanol (1.5 mL) was cooled to 0 to -10°C. Under cooling, 4 mol/L-lithium borohydride-THF solution (0.125 mL, 0.
50 mmol) was added, and the mixture was stirred at the same temperature for 10 minutes. Then 2-((3,4,5-tris(octadecyloxy)benzoyl)oxy)acetic acid N ⁴ -benzoyl-5′-O-(
(2-Cyanoethoxy)(5′-O-(4,4′-dimethoxytrityl)-2′-O-methyluridin-3′-yl)phosphoryl)-2′-O-methylcytidin-3′-yl (0
. 200 g, 0.10 mmol) of THF (0.9 mL)-2-propanol (0.1 mL
After adding the mixed solution of L), the mixture was stirred at the same temperature for 0.5 hour. A 10% ammonium chloride aqueous solution (2 mL) was added to the reaction mixture, and the mixture was separated, and the organic layer was washed with 10% brine. The organic layer was dried over anhydrous sodium sulfate, filtered to remove sodium sulfate, and the filtrate was concentrated. The solid precipitated by adding methanol to the concentrated residue was removed by filtration, and the filtrate was concentrated to dryness. The title compound (0.055 g, 53
%) was obtained as a white solid.
¹ H-NMR (400 MHz, CDCl ₃ ): δ 2.00-2.27 (brs, 2H),
2.55-2.74 (m, 1H), 2.80 (t, 1H, J=6.2Hz), 3.14-
3.41 (m, 1H), 3.44-3.92 (m, 14H), 4.03-4.64 (m,
7H), 5.12 (q, 1H, J = 5.6Hz), 5.27 (t, 1H, J = 8.0Hz)
), 5.87-6.05 (m, 2H), 6.78-6.89 (m, 4H), 7.19-7
. 64 (m, 12H), 7.76-8.16 (m, 4H), 9.05-9.42 (brs
, 1H), 9.50-9.87 (brs, 1H).
¹³ C-NMR (100 MHz, CDCl ₃ ): δ19.54, 19.56, 19.61
, 19.64, 55.28, 55.32, 58.80, 58.82, 58.93, 61.
30, 61.34, 62.38, 62.43, 62.68, 62.73, 66.03, 6
6.08, 66.41, 66.46, 67.85, 67.95, 74.30, 74.34
, 74.42, 74.47, 81.32, 81.41, 81.49, 81.58, 81.
64, 81.72, 82.17, 82.99, 83.08, 86.58, 86.79, 8
7.46, 87.48, 89.42, 89.51, 96.78, 102.69, 102.
71, 113.39, 116.35, 116.48, 127.37, 127.42, 12
7.80, 127.85, 128.12, 128.25, 128.29, 128.57,
128.90, 128.93, 130.22, 130.27, 130.30, 131.0
1, 132.94, 133.02, 133.14, 133.17, 134.72, 134
. 75, 134.82, 134.84, 139.51, 139.66, 143.90, 1
43.97, 144.26, 150.43, 154.73, 158.81, 158.85
, 162.82, 162.86, 163.22, 166.72.
TOF _/ MS ₍ ESI): calcd _{for C51H53N6O16PNa} [M+
Na] ⁺ 1059.3153, found 1059.3202.

[Example 7-8]
5'-O-((2-cyanoethoxy)(5'-O-(4,4'-dimethoxytrityl)
-2′-O-(tert-butyldimethylsilyl)uridin-3′-yl)phosphoryl)
Synthesis of -2'-O-(tert-butyldimethylsilyl)uridine

2-((3,4,5-tris(octadecyloxy)benzoyl)oxy)acetic acid 5′-
O-((2-cyanoethoxy)(5'-O-(4,4'-dimethoxytrityl)-2'-
O-(tert-butyldimethylsilyl)uridin-3'-yl)phosphoryl)-2'-
O-(tert-butyldimethylsilyl)uridin-3′-yl (0.420 g, 0.2
0 mmol) of THF (27 mL)-2-propanol (3 mL) to 0 to -1
Cooled to 0°C. Under cooling, 4 mol / L-lithium borohydride-THF solution (0.2
5 mL, 1.00 mmol) was added, and the mixture was stirred at the same temperature for 1.5 hours. 10 in the reaction liquid
% ammonium chloride aqueous solution (4 mL) was added, and the mixture was separated, and the organic layer was washed with 10% brine. After drying the organic layer with anhydrous sodium sulfate and removing the sodium sulfate by filtration,
The filtrate was concentrated. The solid precipitated by adding methanol to the concentrated residue was removed by filtration, and the filtrate was concentrated to dryness. Column chromatography (spherical neutral silica gel,
Developing solvent: dichloromethane-methanol) to give the title compound (0.1
35 g, 60%) as a white solid.
¹ H-NMR (400 MHz, CDCl ₃ ): δ0.11-0.15 (m, 12H), 0
. 89-0.93 (m, 18H), 1.66-1.84 (brs, 2H), 2.50-2
. 77 (m, 3H), 3.46-3.67 (m, 2H), 3.80 (s, 6H), 3.9
8-4.55 (m, 9H), 4.92-4.96 (m, 1H), 5.23-5.27 (m
, 1H), 5.61-5.75 (m, 2H), 5.97-6.00 (m, 1H), 6.8
4-6.87 (m, 4H), 7.14-7.45 (m, 10H), 7.77-7.87 (
m, 1H), 9.00-9.50 (brs, 2H).
¹³ C-NMR (100 MHz, CDCl ₃ ): δ -5.14, -4.98, -4.96
, -4.90, -4.82, -4.76, -4.74, -4.70, -4.68, 17.
96, 18.02, 18.06, 19.54, 19.60, 19.67, 25.56, 2
5.60, 25.66, 25.69, 55.31, 62.02, 62.29, 62.34
, 62.52, 62.57, 62.62, 67.06, 67.12, 67.32, 69.
38, 69.54, 70.49, 70.93, 73.61, 74.57, 74.63, 7
4.71, 74.88, 81.55, 81.60, 81.73, 81.84, 81.93
, 82.01, 87.37, 87.65, 87.85, 87.95, 88.05, 91.
58, 91.81, 93.18, 93.89, 102.60, 102.67, 102.7
6, 102.82, 102.86, 113.41, 116.26, 116.30, 127
. 41, 127.45, 128.13, 128.17, 128.21, 130.16, 1
30.23, 130.29, 130.34, 134.65, 134.69, 134.72
, 134.75, 139.67, 139.75, 140.36, 140.40, 141.
65, 141.77, 143.99, 144.03, 149.97, 150.09, 15
0.19, 150.40, 150.55, 150.75, 158.83, 158.88,
162.93, 163.00, 163.18.
TOF/MS _{( ESI} ): _{calcd for C54H72N5O16Si2PNa}
[M+Na] ⁺ 1156.4148, found 1156.4148.

5'-O-((2-cyanoethoxy)(5'-O-(4,4'-dimethoxytrityl)
Synthesis of thymidin-3′-yl)phosphoryl)thymidine [Example 7-9]
(1) 3-((3,4,5-tris(octadecyloxy)benzoyl)oxy)propionate 5′-O-((2-cyanoethoxy)(5′-O-(4,4′-dimethoxytrityl) ) thymidin-3′-yl)phosphoryl)thymidin-3′-yl

3-((3,4,5-tris(octadecyloxy)benzoyl)oxy)propionic acid 5′-O-((2-cyanoethoxy)(5′-O-(4
,4′-dimethoxytrityl)thymidin-3′-yl)phosphoryl)thymidin-3′-yl (0.377 g, 0.200 mmol) gave the title compound (0.136 g, 76%) as a white solid.
¹ H-NMR (400 MHz, CDCl ₃ ): δ1.41 (d, 3H, J=5.4 Hz)
, 1.87 (d, 3H, J=2.0Hz), 2.19-2.27 (m, 1H), 2.34
-2.48 (m, 2H), 2.62-2.77 (m, 3H), 3.39 (t, 1H, J =
7.2 Hz), 3.47-3.52 (m, 1H), 3.78 (s, 6H), 4.07-4
. 55 (m, 8H), 5.12-5.18 (m, 1H), 6.18-6.24 (m, 1H
), 6.35-6.40 (m, 1H), 6.84 (d, 4H, J=8.8Hz), 7.2
1-7.31 (m, 8H), 7.32-7.38 (m, 2H), 7.51-7.58 (m
, 1H), 9.92-10.14 (m, 2H).
¹³ C-NMR (100 MHz, CDCl ₃ ): δ 11.78, 12.45, 12.49
, 19.59, 19.67, 19.71, 38.83, 39.01, 39.61, 39.
67, 55.31, 62.45, 62.50, 62.60, 62.65, 63.31, 6
7.74, 70.84, 80.08, 84.40, 84.50, 85.60, 87.30
, 87.36, 111.23, 111.34, 111.91, 112.00, 113.3
7, 116.49, 116.74, 127.32, 128.10, 130.11, 130
. 14, 134.98, 135.01, 135.04, 135.19, 136.00, 1
44.01, 144.06, 150.69, 151.00, 151.05, 158.80
, 163.96, 164.16.
TOF/MS ₍ ESI): _calcd for _{C44H48N5O14PNa} [ _M +
Na] ⁺ 924.2833, found 924.2836.

[Examples 7-10]
(2) 2-(N-methyl-3,4,5-tris(octadecyloxy)benzamide)
Synthesis from 5′-O-((2-cyanoethoxy)(5′-O-(4,4′-dimethoxytrityl)thymidin-3′-yl)phosphoryl)thymidin-3′-yl acetate

2-(N-methyl-3,4,5-tris(octadecyloxy)benzamido)acetic acid 5
'-O-((2-cyanoethoxy)(5'-O-(4,4'-dimethoxytrityl)thymidin-3'-yl)phosphoryl)thymidin-3'-yl (189 mg, 0.10 mmol
) in THF (9 mL)-2-propanol (1 mL) was cooled to 0°C. Under cooling, 4 mol/L-lithium borohydride-THF solution (0.125 mL, 0.50 mm
ol) was added, and the mixture was stirred at the same temperature for 30 minutes. After adding an aqueous solution of ammonium chloride to the reaction solution, extraction was performed three times with dichloromethane. The organic layer was washed with brine, dried over anhydrous sodium sulfate, filtered to remove sodium sulfate, and the filtrate was concentrated. The solid precipitated by suspending the concentrated residue in acetonitrile was removed by filtration, and the filtrate was concentrated.
The concentrated residue was purified by column chromatography (silica gel, developing solvent: chloroform-methanol) to give the title compound (89 mg, 98%) as a white solid.
TOF/MS ₍ ESI): _calcd for _{C44H48N5O14PNa} [ _M +
Na] ⁺ 924.3, found 924.2.

[Example 7-11]
(3) 3-(3,4,5-tris(octadecyloxy)benzamido)propionic acid 5′-O-((2-cyanoethoxy)(5′-O-(4,4′-dimethoxytrityl)thymidine- 3′-yl)phosphoryl)thymidin-3′-yl

3-(3,4,5-tris(octadecyloxy)benzamido)propionic acid 5′-
O-((2-cyanoethoxy)(5′-O-(4,4′-dimethoxytrityl)thymidin-3′-yl)phosphoryl)thymidin-3′-yl (189 mg, 0.10 mmol) in THF (9 mL) )-2-propanol (1 mL) mixed solution was cooled to 0°C. under cooling,
4 mol/L-lithium borohydride-THF solution (0.125 mL, 0.50 mmol
) was added and stirred at room temperature for 1 hour. After adding an aqueous solution of ammonium chloride to the reaction solution, extraction was performed three times with dichloromethane. The organic layer was washed with brine, dried over anhydrous sodium sulfate, filtered to remove sodium sulfate, and the filtrate was concentrated. The solid precipitated by suspending the concentrated residue in acetonitrile was removed by filtration, and the filtrate was concentrated. The concentrated residue was purified by column chromatography (silica gel, developing solvent: chloroform-methanol) to give the title compound (74 mg, 82%) as a white solid.
TOF/MS ₍ ESI): _calcd for _{C44H48N5O14PNa} [ _M +
Na] ⁺ 924.3, found 924.2.

[Example 7-12]
(6-(N ⁶ -benzoyladenin-9-yl)-4-tritylmorpholin-2-yl)methyl P-(2-(hydroxymethyl)-6-thymin-1-yl)morpholino)-N
, N-dimethylphosphonoamidate synthesis

2-((3,4,5-Tris(octadecyloxy)benzoyl)oxy)acetic acid (4-(((6-(N ⁶ -benzoyladenin-9-yl)-4) was prepared according to the method of Example 7-7.
-tritylmorpholin-2-yl)methoxy)(dimethylamino)phosphoryl)-6-(
Thymin-1-yl)morpholin-2-yl)methyl (0.379 g, 0.20 mmol)
to give the title compound (0.114 g, 61%) as a white solid.
¹ H-NMR (400 MHz, CDCl ₃ ): δ 0.87-1.95 (m, 6H),2.
01 (s, 1H), 2.45-2.64 (m, 7H), 3.06-4.04 (m, 9H)
, 4.43-4.46(m, 1H), 5.52(dd, 1H, J=2.4, 10.4Hz
), 6.41 (dt, 1H, J = 2.4, 10.0Hz), 7.05-7.64 (m, 1
9H), 8.03 (d, 2H, J = 8.4Hz), 8.09 (s, 1H), 8.71-8
. 80 (m, 1H), 9.69-9.78 (m, 1H), 10.28 (s, 1H), 10
. 75-11.05 (brs, 1H).
¹³ C-NMR (100 MHz, CDCl ₃ ): δ12.41, 12.45, 36.54
, 36.58, 36.62, 36.66, 44.61, 44, 82, 47.13, 48.
68, 48.78, 52.80, 52.85, 62.72, 62.78, 65.46, 6
5.61, 75.07, 75.14, 75.45, 75.52, 78.06, 78.13
, 78.27, 78.33, 79.60, 79.65, 80.01, 80.49, 110
. 96, 111.03, 122.47, 122.78, 126.65, 128.00, 1
28.24, 128.41, 128.49, 128.62, 129.17, 132.61
, 132.69, 133.48, 133.64, 134.86, 134.96, 140.
94, 149.54, 149.75, 149.82, 150.02, 151.23, 15
1.42, 152.66, 152.75, 163.75, 163.79, 165.25.
TOF _/ MS (ESI): calcd _{for C48H51N10O8PNa [} M+
Na] ⁺ 949.3527, found 949.3545.

[Example 7-13]
(6-(N ⁴ -benzoylcytosin-1-yl)-4-tritylmorpholin-2-yl)methyl P-(2-(hydroxymethyl)-6-(N ² -isobutyrylguanin-9-yl) Synthesis of morpholino)-N,N-dimethylphosphonoamidate

2-((3,4,5-Tris(octadecyloxy)benzoyl)oxy)acetic acid (4-(((6-(N ⁴ -benzoylcytosin-1-yl)-4) was prepared according to the method of Example 7-7.
-tritylmorpholin-2-yl)methoxy)(dimethylamino)phosphoryl)-6-(
N ² -isobutyrylguanin-9-yl)morpholin-2-yl)methyl (0.197 g
, 0.10 mmol) gave the title compound (0.052 g, 52%) as a white solid.
¹ H-NMR (400 MHz, CDCl ₃ ): δ 0.78-1.55 (m, 9H),2.
37-2.95 (m, 8H), 3.08-3.94 (m, 9H), 4.13-4.43 (
m, 2H), 5.42-5.54 (m, 1H), 6.22-6.38 (m, 1H), 7.
05-7.80 (m, 19H), 7.93 (d, 2H, J=7.2Hz), 9.58-9
. 92 (brs, 1H), 10.64-10.86 (brs, 1H), 12.05-12
. 21 (brs, 1H).
¹³ C-NMR (100 MHz, CDCl ₃ ): δ 13.91, 14.12, 19.03
, 19.06, 19.28, 22.69, 25.76, 28.55, 29.36, 29.
70, 31.72, 31.92, 35.99, 36.05, 36.62, 36.65, 3
6.69, 44.63, 44.92, 47.15, 47.56, 48.44, 48.79
, 52.26, 52.42, 61.85, 62.58, 62.73, 65.66, 66.
15, 71.09, 71.69, 75.40, 75.86, 75.91, 78.19, 7
9.73, 80.56, 81.37, 81.76, 97.87, 120.56, 120.
92, 126.65, 127.96, 128.04, 128.70, 128.83, 12
8.92, 129.13, 131.00, 132.21, 132.97, 133.02,
133.14, 136.66, 137.43, 144.02, 144.54, 147.9
2, 147.98, 148.00, 154.99, 155.57, 155.62, 162
. 69, 163.09, 167.24, 179.62, 179.86.
TOF/MS ( _ESI ): _{calcd for C51H56N11O9PNa} [M+
Na] ⁺ 1020.3898, found 1020.3942.

[Example 7-14]
Confirmation of 8-mer detagging (1) 2-((3,4,5-tris(octadecyloxy)benzoyl)oxy)acetic acid 5′-O-((2-cyanoethoxy)(5′-O-( (2-cyanoethoxy)(5′-O
-((2-cyanoethoxy)(5'-O-((2-cyanoethoxy)(5'-O-((2
-cyanoethoxy)(5′-O-((2-cyanoethoxy)(5′-O-((2-cyanoethoxy)(N ⁴ -benzoyldeoxycytidine-3′-yl)phosphoryl)-N ⁶ -benzoyl deoxyadenosin-3′-yl)phosphoryl)thymidin-3′-yl)phosphoryl)-N ² -isobutyryldeoxyguanosin-3′-yl)phosphoryl)-N ⁴ -benzoyldeoxycytidin-3′-yl)phosphoryl )-N ⁶ -benzoyldeoxyadenosin-3′-yl)phosphoryl)thymidin-3′-yl)phosphoryl)thymidine-3′
-yl (8mer form) Under argon, 2-((3,4,5-tris(octadecyloxy)benzoyl)oxy)thymidin-3′-yl acetate (1.19 g, 0.98 mmol), 5′ -O-(4,4
'-Dimethoxytrityl)thymidin-3'-yl-phosphoramidite (1.46 g, 1
. 96 mmol) in dichloromethane (10 mL) was added with 5-benzylthio-1H-tetrazole (0.38 g, 1.96 mmol) and stirred at room temperature for 1.5 hours.
Then pyridine (0.93 g, 11.77 mmol) and water (0.11 g, 5.89 mmol)
l), iodine (1.25 g, 4.91 mmol) was added, and the mixture was stirred at room temperature for 4 hours.
After adding 1.2 mol/L-ascorbic acid aqueous solution (3 mL) to the reaction solution,
Stir for a minute. Acetonitrile (24 mL) was added dropwise to the reaction solution, and the precipitated solid was filtered. The solid was washed with an acetonitrile-dichloromethane mixed solvent and then dried under reduced pressure at 50° C. to obtain a DMTr form (2.22 g).

Next, a suspension of DMTr form (1.83 g, 0.98 mmol) in dichloromethane (15 mL
L), pyrrole (0.33 g, 4.91 mmol), trifluoroacetic acid (0.13 g, 1
. 18 mmol) was added, and the mixture was stirred at room temperature for 1.5 hours. Acetonitrile (
37 mL) and methanol (18 mL) were added dropwise, and the precipitated solid was filtered. The solid was washed with a mixed solvent of acetonitrile-methanol-dichloromethane and then dried under reduced pressure at 50° C. to obtain a 2mer compound (1.48 g).

An operation similar to the above was repeated six times to obtain an 8-mer (2.94 g).
TOF/MS (ESI): m/z 2106.3 [M+2H] ^<2+> , 1403.9
[M+3H] ³⁺ .

[Example 7-15]
(2) 2-((3,4,5-tris(octadecyloxy)benzoyl)oxy)acetic acid 5′-O-((2-cyanoethoxy)(5′-O-((2-cyanoethoxy)(5 '-O
-((2-cyanoethoxy)(5'-O-((2-cyanoethoxy)(5'-O-((2
-cyanoethoxy)(5′-O-((2-cyanoethoxy)(5′-O-((2-cyanoethoxy)(N ⁴ -benzoyldeoxycytidine-3′-yl)phosphoryl)-N ⁶ -benzoyl deoxyadenosin-3′-yl)phosphoryl)thymidin-3′-yl)phosphoryl)-N ² -isobutyryldeoxyguanosin-3′-yl)phosphoryl)-N ⁴ -benzoyldeoxycytidin-3′-yl)phosphoryl )-N ⁶ -benzoyldeoxyadenosin-3′-yl)phosphoryl)thymidin-3′-yl)phosphoryl)thymidine-3′
-Detag of yl N,N-diisopropylethylamine (65 mg, 0.50 mmol) was added to a mixed solution of dichloromethane (0.4 mL), THF (0.2 mL), and 2-propanol (0.04 mL). Cooled to -10°C. Next, 4 mol / L-lithium borohydride-TH
After adding F solution (0.125 mL, 0.50 mmol), the mixture was stirred at the same temperature for 20 minutes,
A reducing reagent was prepared. The 8-mer (84 mg, 0.25 mg, 0.25 mg,
02 mmol) of dichloromethane (4.2 mL)-THF (1.8 mL)-2-propanol (0.4 mL) mixture was added with the reducing reagent and stirred at the same temperature for 40 minutes. 1 in reaction liquid
After adding 0% aqueous ammonium chloride solution (5 mL), the mixture was extracted with dichloromethane. After washing the organic layer with brine, the organic layer was concentrated and dried. The solid precipitated by suspending the concentrated residue in dichloromethane was collected by filtration. After washing the solid with dichloromethane, it is dried under reduced pressure, and 5'-O-((2-cyanoethoxy)(5'-O-((2-cyanoethoxy)(5'
-O-((2-cyanoethoxy)(5'-O-((2-cyanoethoxy)(5'-O-(
(2-cyanoethoxy)(5′-O-((2-cyanoethoxy)(5′-O-((2-cyanoethoxy)(N ⁴ -benzoyldeoxycytidin-3′-yl)phosphoryl)-N ⁶
-benzoyldeoxyadenosin-3′-yl)phosphoryl)thymidin-3′-yl)phosphoryl)-N ² -isobutyryldeoxyguanosin-3′-yl)phosphoryl)-N ⁴
-benzoyldeoxycytidin-3′-yl)phosphoryl)-N ⁶ -benzoyldeoxyadenosin-3′-yl)phosphoryl)thymidin-3′-yl)phosphoryl)thymidine (
25 mg) was obtained.
TOF/MS (ESI): m/z 1621.9 [M+2H] ^<2+> , 1081.6
[M+3H] ³⁺ , 811.2 [M+4H] ⁴⁺ .

8. Comparison of detagging progress rate [Example 8]
2-((3,4,5-tris(octadecyloxy)benzoyl)oxy)acetic acid 5′-
O-(4,4'-dimethoxytrityl)thymidin-3'-yl (A-Tag-dT-DM
Tr) and 3,4,5-tris(octadecyloxy)benzyl succinate 5′-O-
(4,4′-dimethoxytrityl)thymidin-3′-yl (S-Tag-dT-DMTr
(abbreviated as )) were compared in terms of the rate of progress of detagging under the following ammoniacal basic conditions.

<Detagging conditions>
A-Tag-dT-DMTr or S-Tag-dT-DMTr 50 μmol
28% ammonia water 2.0 mL
ethanol 0.5 mL
THF 0.5 mL
Reaction temperature 35°C

<Production rate of dT-DMTr (untagged)>

<HPLC conditions>
Column: Inertsil WP300 Diol (5 μm, 4.6Φ×150 mm
)
Flow rate: 1.0 mL/min
Mobile phase: n-hexane, CHCl ₃ -MeOH (1/1)
Gradient: 0-11 min; 5 to 30% ( _CHCl3
-MeOH)
Detection method: UV (λ = 254 nm)
Retention time: A-Tag-dT-DMTr: Rt = 4.23 min, S-Tag-dT-
DMTr: Rt = 4.17 min, dT-DMTr: Rt = 9.40 min
Production rate: (dT-DMTr) / [(A-Tag-dT-DMTr or S-Tag-
dT-DMTr) + (dT-DMTr)]

As described above, it was confirmed that A-Tag is detagged more rapidly than the succinate under general deprotection conditions for oligonucleic acid synthesis.

9. Summary of Synthesis Examples A summary of synthesis examples in each of the above-described examples is shown below.

9.1 Synthesis of trimers

As shown in Table 2 above, various Tag-bound trimeric DNA oligomers could be obtained in high yields.

9.2 Dimer Synthesis

As shown in Table 3 above, various Tag-linked oligonucleotides (dimers) could be obtained in high yields.

9.3 Dimer Synthesis

As shown in Table 4 above, various Tag-linked oligonucleotides (dimers) could be obtained in high yields.

9.4 Dimer Synthesis

As shown in Table 5 above, various Tag-linked oligonucleotides (dimers) could be obtained in high yields.

9.5 Synthesis of 20-mer and Total Deprotection

By using the alkoxyphenyl derivative and tagged nucleotide protected body of the present invention, the tagged oligonucleotide protected body (20-mer) could be easily obtained. Furthermore, the oligonucleotide could be easily obtained by removing the Tag portion and the protecting group.

9.6 Trimer Detag

As shown in Table 6 above, T of various tagged oligonucleotide protected bodies (trimers)
By removing the ag portion, each protected oligonucleotide (trimer) could be obtained in high yield.

9.7 Detagging Dimers

As shown in Table 7 above, T of various tagged oligonucleotide protected bodies (dimers)
By removing the ag portion, each protected oligonucleotide (dimer) could be obtained in high yield.

9.8 Detagging Dimers

As shown in Table 8 above, the T of various tagged oligonucleotide protected bodies (dimers)
By removing the ag portion, each protected oligonucleotide (dimer) could be obtained in high yield.

9.9 Detagging Dimers

As shown in Table 9 above, the T of various tagged oligonucleotide protected bodies (dimers)
By removing the ag portion, each protected oligonucleotide (dimer) could be obtained in high yield.

The alkoxyphenyl derivative of the present invention is the above tagged nucleoside protected body and Tag
It can be used for the production of protected nucleotides. In addition, the method for producing an oligonucleotide such as a protected nucleoside and a protected nucleotide of the present invention, and a method for selectively removing a Tag portion of a protected nucleoside or a protected nucleotide of the present invention are particularly effective in a liquid phase. It can be used for the synthesis of oligonucleotide protectors in synthesis (including pseudo-liquid phase synthesis). All of these are extremely useful compounds and techniques for development of nucleic acid medicines, elucidation of biological mechanisms, and the like.

Claims (18)

A tagged mono- or oligonucleoside or a tagged mono- or oligonucleotide having a Tag portion shown in the partial structure below.

(In the formula,
Each R independently represents a linear or branched alkyl group having 10 to 40 carbon atoms.
m represents an integer of 1 to 5; When m is 2 or more, multiple ROs may be the same or different.
X represents O, S, NH, or ^NRN .
n represents an integer of 1-4.
R ^N represents an optionally substituted alkyl group having 1 to 6 carbon atoms.
* represents the point of attachment to a mono- or oligonucleoside or mono- or oligonucleotide,
The partial structure is bound to the oxygen atom that replaces the carbon atom located at the 3'-position of the nucleoside or nucleotide, or the oxygen that replaces the carbon atom located at the 5'-position of the morpholinonucleoside or morpholinonucleotide. Bonded to an atom. ) 2. The mono- or oligonucleoside or mono- or oligonucleotide according to claim 1, wherein said partial structure is represented by the following formula.

(In the formula,
R ¹ , R ² and R ³ each independently represent a linear or branched alkyl group having 10 to 40 carbon atoms.
X represents O, S, NH, or ^NRN .
n represents an integer of 1-4.
R ^N represents an optionally substituted alkyl group having 1 to 6 carbon atoms. ) 3. The mono- or oligonucleoside or mono- or oligonucleotide according to claim 1 or 2, wherein said partial structure is bound to the oxygen atom substituting the carbon atom located at the 3'-position of the nucleoside or nucleotide. 3. The mono- or oligonucleoside or mono- or oligonucleotide according to claim 1 or 2, wherein the partial structure is bound to the oxygen atom substituting the carbon atom located at the 5'-position of the morpholinonucleoside or morpholinonucleotide. A mono- or oligonucleoside or mono- or oligonucleotide according to any one of claims 1 to 3, wherein said nucleoside or nucleotide has a phosphate or phosphorothioate moiety. The mono- or oligonucleoside according to any one of claims 1 to 5, wherein the nucleoside or nucleotide contains an adenyl group, a guanyl group, a citidyl group, a thymidyl group, or a uracil group as Base. or mono- or oligonucleotides. The mono- or oligonucleoside or mono- or oligonucleotide according to any one of claims 1 to 6, wherein n is 1 or 2. The mono- or oligonucleoside or mono- or oligonucleotide according to any one of claims 1, 3-7, wherein m is an integer of 2-4. A method for producing a mono- or oligonucleoside or mono- or oligonucleotide, comprising a step of reducing the tagged mono- or oligonucleoside or the tagged mono- or oligonucleotide according to any one of claims 1 to 8 . 10. The production method according to claim 9, wherein the reduction treatment uses a boron-containing reducing agent or uses a boron-containing reducing agent and an amine. 11. The production method according to claim 10 , wherein the boron-containing reducing agent is lithium borohydride, sodium borohydride, lithium triethylborohydride, or tetrabutylammonium borohydride. A step of reducing the tagged mono- or oligonucleoside or tagged mono- or oligonucleotide according to any one of claims 1 to 8,
A method for removing the Tag portion. 13. The removal method of claim 12, wherein the reduction treatment uses a boron-containing reducing agent or uses a boron-containing reducing agent and an amine. 14. The removal method of claim 13 , wherein the boron-containing reducing agent is lithium borohydride, sodium borohydride, lithium triethylborohydride, tetrabutylammonium borohydride. A tagging agent having a tag portion represented by the partial structure below.

(In the formula,
Each R independently represents a linear or branched alkyl group having 10 to 40 carbon atoms.
m represents an integer of 1 to 5; When m is 2 or more, multiple ROs may be the same or different.
X represents O, S, NH, or ^NRN .
n represents an integer of 1-4.
R ^N represents an optionally substituted alkyl group having 1 to 6 carbon atoms.
* represents a bonding site with a leaving group, and the leaving group is a halogen, an imidazole group, or an alkoxy group. ) 16. The tagging agent according to claim 15, wherein the partial structure is represented by the following formula.

(In the formula,
R ¹ , R ² and R ³ each independently represent a linear or branched alkyl group having 10 to 40 carbon atoms.
X represents O, S, NH, or ^NRN .
n represents an integer of 1-4.
R ^N represents an optionally substituted alkyl group having 1 to 6 carbon atoms.
* represents a bonding site with a leaving group. ) 17. The tagging agent according to claim 15 or 16, wherein said n is 1 or 2. The tagging agent according to any one of claims 15 to 17, which is used for tagging mono- or oligonucleosides or mono- or oligonucleotides.