Lipid-binding oligonucleotides or their complexes

Abstract

The objective is to provide lipid-binding oligonucleotides or their complexes that enhance pharmacological effects in the liver and other organs. [Solution] The present invention provides a lipid-binding oligonucleotide or a complex thereof, represented by the following general formula (I), or a pharmaceutical composition containing the same. TIFF2026131055000022.tif18165 (In the formula, W is a group derived from an oligonucleotide compound or oligonucleotide complex, L is a divalent group derived from a substituted or unsubstituted C1-10 alkylene group or a substituted or unsubstituted polyC1-10 alkylene glycol, and R1 and R2 are each independently a substituted or unsubstituted C5-32 alkyl group or a substituted or unsubstituted C5-32 alkenyl group, or R1 and R2 may be bonded to each other to form a ring.)

Description

Technical Field

[0001] The present invention relates to a lipid-conjugated oligonucleotide or a complex thereof.

Background Art

[0002] Nucleic acid drugs are drugs composed of nucleic acids (oligonucleotides) that form complementary base pairs with target DNA or RNA, and are expected as novel drugs. Representative nucleic acid drugs include antisense nucleic acids (ASO) and siRNA, etc. However, developing means for efficiently delivering these to target organs, particularly to organs other than the liver, has long been recognized as a problem.

[0003] As a delivery means, it has been reported that when lipids such as cholesterol, tocopherol, or palmitic acid are conjugated to ASO, the antisense effect is improved in muscles and the heart (see, for example, Non-Patent Document 1). However, the effect is limited. Also, it has been shown that toxicity occurs with cholesterol conjugates.

[0004] Furthermore, it has also been reported that a complex composed of a first oligonucleotide containing ASO and a second oligonucleotide conjugated with a lipid such as cholesterol improves the antisense effect on organs other than the liver (see, for example, Patent Document 1).

[0005] Thus, it has been shown that by conjugating a lipid to an oligonucleotide, the effect can be exerted on organs other than the liver, but many of the effects are limited, and there is still a desire to develop more effective lipids as delivery means.

Prior Art Documents

Patent Documents

[0006]

Patent Document 1

Non-Patent Documents

[0007] [Non-Patent Document 1] Nucleic Acids Research, 2019, 47, 12, pp 6045-6058 [Overview of the project] [Problems that the invention aims to solve]

[0008] The object of this invention is to provide lipid-binding oligonucleotides or complexes thereof that improve pharmacological effects in the liver and other organs. [Means for solving the problem]

[0009] To solve the above problems, the inventors conducted extensive research and found that oligonucleotides or complexes thereof conjugated with novel dialkyl lipids improve their pharmacological effects on the liver and other organs. Based on these findings, the inventors have completed the present invention. That is, the present invention encompasses the following aspects.

[0010] [1] A lipid-binding oligonucleotide or its complex, represented by the following general formula (I). [ka] (In the formula, W is a group derived from an oligonucleotide compound or oligonucleotide complex. L is a divalent group derived from a substituted or unsubstituted C1-10 alkylene group or a substituted or unsubstituted polyC1-10 alkylene glycol. R1 and R2 may each be independently a substituted or unsubstituted C5-32 alkyl group or a substituted or unsubstituted C5-32 alkenyl group, or R1 and R2 may be bonded to each other to form a ring. [2] The lipid-binding oligonucleotide or complex thereof according to [1], wherein R1 and R2 are each independently an unsubstituted C5-32 alkyl group or an unsubstituted C5-32 alkenyl group, or R1 and R2 are bonded to each other to form a ring. [3] The lipid-binding oligonucleotide or complex thereof according to [1] or [2], wherein R1 and R2 are each independently an unsubstituted C5-32 alkyl group or an unsubstituted C5-32 alkenyl group. [4] A lipid-binding oligonucleotide or complex thereof according to any one of [1] to [3], wherein R1 and R2 are each independently an unsubstituted C10-20 alkyl group. [5] A lipid-binding oligonucleotide or complex thereof according to any one of [1] to [4], wherein R1 and R2 are unsubstituted C14 alkyl groups.

[0011] [6] A lipid-binding oligonucleotide or complex thereof according to any one of [1] to [5], wherein L is an unsubstituted C1-10 alkylene group. [7] The lipid-binding oligonucleotide or complex thereof according to any one of [1] to [6], wherein L is an unsubstituted C6 alkylene group. [8] The lipid-binding oligonucleotide according to any one of [1] to [7], wherein the oligonucleotide compound is a gapmer-type antisense oligonucleotide. [9] The lipid-binding oligonucleotide according to [8], wherein the gapmer-type antisense oligonucleotide comprises at least one selected from the group consisting of a 2′-modified non-crosslinked nucleoside and a 2′-4′-crosslinked nucleoside.

[10] The lipid-binding oligonucleotide according to [8] or [9], wherein the gapmer-type antisense oligonucleotide comprises at least four consecutive deoxyribonucleosides.

[0012]

[11] The lipid-conjugated oligonucleotide according to any one of [8] to

[10] , wherein the gapmer-type antisense oligonucleotide consists of 13 to 25 nucleosides.

[12] L is the lipid-conjugated oligonucleotide according to any one of [8] to

[11] , which is bound to the 5'-end of the gapmer-type antisense oligonucleotide.

[13] The lipid-conjugated oligonucleotide according to any one of [1] to [7], wherein the oligonucleotide compound is a mixmer-type antisense oligonucleotide.

[14] The lipid-conjugated oligonucleotide according to

[13] , wherein the mixmer-type antisense oligonucleotide contains at least one selected from the group consisting of 2'-modified non-bridged nucleotides and 2'-4'-bridged nucleotides.

[15] The lipid-conjugated oligonucleotide according to

[13] or

[14] , wherein the mixmer-type antisense oligonucleotide is an oligonucleotide in which a nucleoside or oligonucleotide consisting of 1 to 20 sugar-modified nucleosides and a nucleoside or oligonucleotide consisting of 1 to 3 deoxyribonucleosides are alternately linked, or an oligonucleotide composed only of sugar-modified nucleosides as nucleosides.

[0013]

[16] The oligonucleotide compound according to any one of

[13] to

[15] , wherein the mixmer-type antisense oligonucleotide consists of 13 to 25 nucleosides.

[17] L is the lipid-conjugated oligonucleotide compound according to any one of

[13] to

[16] , which is bound to the 5'-end of the mixmer-type antisense oligonucleotide.

[18] The oligonucleotide complex is a double-stranded oligonucleotide complex comprising a first oligonucleotide and a second oligonucleotide, The first oligonucleotide is a gapmer-type or mixed-type antisense oligonucleotide consisting of 7 to 100 nucleosides. The second oligonucleotide contains a sequence that enables hybridization with at least a portion of the first oligonucleotide, and consists of 4 to 100 nucleosides independently selected from deoxyribonucleosides, ribonucleosides, and sugar-modified nucleosides, and the first or second oligonucleotide is bound to L. A complex according to any one of [1] to [7], wherein a primary oligonucleotide and a secondary oligonucleotide are hybridized.

[19] The complex according to

[18] , wherein the second oligonucleotide comprises at least four consecutive ribonucleosides.

[20] The complex according to

[18] or

[19] , wherein L is bound to the 5′ end of the second oligonucleotide.

[0014] [twenty one] The oligonucleotide compound comprises a first oligonucleotide and a second oligonucleotide, The first oligonucleotide is a gapmer-type or mixed-type antisense oligonucleotide consisting of 7 to 100 nucleosides. The second oligonucleotide contains a sequence that enables hybridization with at least a portion of the first oligonucleotide, and consists of 4 to 100 nucleosides independently selected from deoxyribonucleosides, ribonucleosides, and sugar-modified nucleosides, and the first or second oligonucleotide is bound to L. The first oligonucleotide and the second oligonucleotide are linked together. A lipid-binding oligonucleotide as described in any of [1] to [7], wherein the first oligonucleotide and the second oligonucleotide are hybridized. [twenty two] The lipid-binding oligonucleotide according to

[21] , wherein the second oligonucleotide comprises at least four consecutive ribonucleosides. [twenty three] The lipid-binding oligonucleotide according to

[21] or

[22] , wherein L is bound to the 5′ end of the second oligonucleotide. [twenty four] The lipid-binding oligonucleotide according to any one of [1] to [7], wherein the oligonucleotide compound or oligonucleotide complex is selected from siRNA, aptamer, and ribozyme. [twenty five] A pharmaceutical composition comprising a lipid-binding oligonucleotide or a complex thereof as described in any of [1] to

[24] , and a pharmacokinetically acceptable carrier.

[26] A method for controlling the function of a target RNA, comprising the step of contacting a cell with a lipid-binding oligonucleotide or a complex thereof described in any of [1] to

[24] .

[27] A method for controlling the function of a target RNA in a mammal, comprising the step of administering the pharmaceutical composition described in

[25] to the mammal.

[28] A method for controlling the expression of a target gene, comprising the step of contacting a cell with a lipid-binding oligonucleotide or a complex thereof described in any of [1] to

[24] .

[29] A method for controlling the expression of a target gene in a mammal, comprising the step of administering the pharmaceutical composition described in

[25] to the mammal.

[30] A compound represented by the following formula (II). [ka] (In the formula, L is a divalent group derived from a substituted or unsubstituted C1-10 alkylene group or a substituted or unsubstituted polyC1-10 alkylene glycol. R1 and R2 may each be independently a substituted or unsubstituted C5-32 alkyl group or a substituted or unsubstituted C5-32 alkenyl group, or R1 and R2 may be bonded to each other to form a ring. Each of the R3 groups may be independently a C1-10 alkyl group, or the two alkyl groups of R3 may be bonded to each other to form a ring. R4 is a 2-cyanoethyl group. [Effects of the Invention]

[0015] The present invention provides lipid-binding oligonucleotides or complexes thereof with enhanced pharmacological effects on the liver and other organs. [Brief explanation of the drawing]

[0016] [Figure 1] This graph shows the results of Malat1 expression levels 10 days after administration of antisense oligonucleotides in mice, as demonstrated in Evaluation Example 3, regarding antisense suppression of Malat1. [Figure 2] This graph shows the results of Malat1 expression levels 3 days after administration of antisense oligonucleotides in mice, as demonstrated in Evaluation Example 3, regarding antisense suppression of Malat1. [Figure 3] This graph shows the results of the HPrt1 expression level in the liver after administration of 2.9 μmol / kg of antisense oligonucleotide in mice, as in Evaluation Example 4, regarding the antisense suppression of HPrt1. [Figure 4] This graph shows the results of the antisense suppression of HPrt1 in mice in Evaluation Example 4, specifically the HPrt1 expression level in the liver 10 days after administration of antisense oligonucleotides. [Figure 5]This graph shows the results of the HPrt1 expression level in the heart of mice administered 2.9 μmol / kg of antisense oligonucleotide in the antisense suppression of HPrt1 in evaluation example 4. [Figure 6] This graph shows the results of the antisense suppression of HPrt1 in mice in Evaluation Example 4, specifically the HPrt1 expression level in the heart 10 days after administration of antisense oligonucleotides. [Figure 7] This graph shows the results of HPrt1 expression levels in the lungs after administration of 2.9 μmol / kg of antisense oligonucleotide in mice, as in Evaluation Example 4, regarding antisense suppression of HPrt1. [Figure 8] This graph shows the results of the antisense suppression of HPrt1 in mice in Evaluation Example 4, specifically the HPrt1 expression level in muscle after administration of 2.9 μmol / kg of antisense oligonucleotide. [Figure 9] This graph shows the results of the antisense suppression of HPrt1 in mice in Evaluation Example 4, specifically the HPrt1 expression level in the muscle 10 days after administration of antisense oligonucleotides. [Modes for carrying out the invention]

[0017] <Terminology> Unless otherwise specified, the terms used in this invention have the following definitions.

[0018] "Nucleoside" is a term well known to those skilled in the art and is generally understood to be a molecule in which a sugar and a nucleic acid base are bonded, and which can be a constituent unit of a nucleic acid. In this specification, nucleoside is a broader concept and includes deoxyribonucleosides, ribonucleosides, and sugar-modified nucleosides, which are described below. The nucleic acid base may be modified.

[0019] "Deoxyribonucleoside" refers to a molecule having a nucleic acid base at the carbon atom at position 1 of 2-deoxyribose. In this invention, the deoxyribonucleoside may be a naturally occurring deoxyribonucleoside or a deoxyribonucleoside in which the nucleic acid base portion of a naturally occurring deoxyribonucleoside has been modified. A naturally occurring deoxyribonucleoside is a deoxyribonucleoside that has a naturally occurring nucleic acid base. Modifications may be applied to a single deoxyribonucleoside in combination of multiple types. The modified deoxyribonucleosides are described, for example, in Journal of Medicinal Chemistry, 2016, 59, pp 9645-9667, Medicinal Chemistry Communication, 2014, 5, pp 1454-1471, Future Medicinal Chemistry, 2011, 3, pp 339-365, and International Publication No. 2018 / 155450, among others.

[0020] "Ribonucleoside" refers to a molecule having a nucleic acid base at the carbon atom at position 1 of ribose. In this invention, the ribonucleoside may be a naturally occurring ribonucleoside or a ribonucleoside in which the nucleic acid base portion of a naturally occurring ribonucleoside has been modified. A naturally occurring ribonucleoside is a ribonucleoside that has a naturally occurring nucleic acid base. Modification may be applied to a single ribonucleoside in combination of multiple types. The aforementioned modified ribonucleosides are described, for example, in Journal of Medicinal Chemistry, 2016, 59, pp 9645-9667, Medicinal Chemistry Communication, 2014, 5, pp 1454-1471, Future Medicinal Chemistry, 2011, 3, pp 339-365, and International Publication No. 2018 / 155450, among others.

[0021] "Modified sugars" are (Z1) A molecule in which ribose or 2-deoxyribose is partially substituted by one or more substituents. (Z2) Molecules in which ribose or 2-deoxyribose is substituted with a penta-monosaccharide or hexa-monosaccharide different from ribose and 2-deoxyribose (e.g., hexitol, threose, etc.), (Z3) Ribose or 2-deoxyribose as a whole, or molecules in which the tetrahydrofuran ring thereof is replaced with a 5- to 7-membered saturated or unsaturated ring (e.g., cyclohexane, cyclohexene, morpholine, etc.), or a substructure (e.g., a peptide structure) that can form a 5- to 7-membered ring by hydrogen bonding. or (Z4) Molecules in which ribose or 2-deoxyribose is replaced with C2-6 alkylene glycol (e.g., ethylene glycol, propylene glycol, etc.) It means... Modified sugars include the "2-modified sugars" and "2-4 cross-linked sugars" described below. Examples of modified sugars and sugar-modified nucleosides described later include sugars and sugar-modified nucleosides disclosed as suitably used in antisense methods in Japanese Patent Publication No. 10-304889, International Publication No. 2005 / 021570, Japanese Patent Publication No. 10-195098, Japanese Patent Publication No. 2002-521310, International Publication No. 2007 / 143315, International Publication No. 2008 / 043753, International Publication No. 2008 / 029619, International Publication No. 2008 / 049085, and International Publication No. 2017 / 142054 (hereinafter, these documents are referred to as "documents relating to antisense methods"), etc. Modified sugars and sugar-modified nucleosides are also disclosed in Journal of Medicinal Chemistry, 2016, 59, pp 9645-9667, Medicinal Chemistry Communication, 2014, 5, pp 1454-1471, Future Medicinal Chemistry, 2011, 3, pp 339-365, and International Publication No. 2018 / 155450, among others.

[0022] Examples of modified sugars partially substituted by a single substituent include ribose or 2-deoxyribose in which any position of the sugar moiety is substituted with (i) or (ii) below. (i) C1-6 alkyl group. Here, if substitution by two or more C1-6 alkyl groups is included, the two or more C1-6 alkyl groups may together form a 3- to 6-membered ring. (ii) A C1-6 alkyl group substituted with at least one selected from the group consisting of a halogen atom, a C1-6 alkoxy group, a halo-C1-6 alkoxy group, a mono- or di-C1-6 alkylamino group, a 5-10 membered heterocyclic group, a carboxyl group, a carbamoyl group, and an N-substituted carbamoyl group. Here, examples of the N-substituted carbamoyl group include the N-methyl-carbamoyl group and the N-ethyl-carbamoyl group, where the methyl and ethyl groups of the N-methyl-carbamoyl group and the N-ethyl-carbamoyl group may be substituted with a 5-10 membered heterocyclic group or a mono- or di-C1-6 alkylamino group. Specific examples of the N-substituted carbamoyl group include the N-methylcarbamoyl group, the N-ethylcarbamoyl group, the N-dimethylaminoethyl-carbamoyl group, the N-morpholinoethylcarbamoyl group, the N-(2-pyridylethyl)carbamoyl group, and the N-((benzimidazole-1-yl)ethyl)carbamoyl group.

[0023] "Sugar-modified nucleoside" refers to a molecule having the aforementioned "modified sugar" instead of the sugar portion of a deoxyribonucleoside or ribonucleoside. For example, it includes the "2'-modified nucleoside" and "2'-4'-bridged nucleoside" described later. If the modified sugar is (Z3) as defined above, sugar-modified nucleoside also includes molecules in which the modified sugar and a nucleic acid base are linked via a methylene chain or the like.

[0024] "2-modified sugars" refer to non-crosslinked sugars in which the oxygen or carbon atom at the 2nd position of ribose is modified, and include "2-O-Me", "2-O-MOE", "2-O-MCE", "2-O-NMA", "2-O-AP", "2-F", "2-DMAECE", "2-MоrECE", "2-PyECE", and "2-BimECE". "2'-modified nucleoside" refers to a molecule having a nucleic acid base at position 1 of the aforementioned 2'-modified sugar, and examples include "2'-O-Me nucleoside", "2'-O-MOE nucleoside", "2'-O-MCE nucleoside", "2'-O-NMA nucleoside", "2'-O-AP nucleoside", "2'-F nucleoside", "2'-DMAECE nucleoside", "2'-MоrECE nucleoside", "2'-PyECE nucleoside", and "2'-BimECE nucleoside".

[0025] "2-O-Me" (also called 2-O-methyl) refers to a sugar in which the hydroxyl group at the 2nd position of ribose is replaced with a methoxy group. A "2'-O-Me nucleoside" (also called a 2'-O-methyl nucleoside) refers to a molecule that has a nucleic acid base at position 1 of "2'O-Me".

[0026] "2-O-MOE" (also known as 2-O-methoxyethyl) refers to a sugar in which the hydroxyl group at position 2 of ribose is replaced by a 2-methoxyethyloxy group. A "2'-O-MOE nucleoside" (also called a 2'-O-methoxyethyl nucleoside) refers to a molecule that has a nucleic acid base at position 1 of the "2'-O-MOE" structure.

[0027] "2-O-MCE" (also known as 2-O-methylcarbamoylethyl) refers to a sugar in which the hydroxyl group at the 2nd position of ribose is replaced by a methylcarbamoylethyloxy group. A "2'-O-MCE nucleoside" (also known as a 2'-O-methylcarbamoylethyl nucleoside) refers to a molecule that has a nucleic acid base at position 1 of "2'-O-MCE".

[0028] "2-O-NMA" refers to a sugar in which the hydroxyl group at position 2 of ribose is replaced by a 2-[(methylamino)-2-oxoethyl]oxy group. A "2'-O-NMA nucleoside" refers to a molecule that has a nucleic acid base at position 1 of the "2'-O-NMA" structure.

[0029] "2-O-AP" refers to a sugar in which the hydroxyl group at position 2 of ribose is replaced with a 3-aminopropyloxy group. "2'-O-AP nucleoside" refers to a molecule that has a nucleic acid base at position 1 of "2'-O-AP".

[0030] "2-F" refers to a sugar in which the hydroxyl group at position 2 of ribose is replaced by a fluorine atom. A "2'-F nucleoside" refers to a molecule that has a nucleic acid base at the 1st position of the "2-F" group.

[0031] "2-DMAECE," "2-MorECE," "2-PyECE," and "2-BimECE" are modified sugars in which the hydroxyl group at position 2 of ribose is replaced with the structures shown below for DMAECE, MorECE, PyECE, and BimECE, respectively. In the following structures, the wavy lines indicate the bonding position of the hydroxyl group at position 2 of ribose to the carbon atom to which it is attached. [ka]

[0032] "2'-DMAECE nucleoside", "2'-MorECE nucleoside", "2'-PyECE nucleoside", and "2'-BimECE nucleoside" refer to molecules that have a nucleic acid base at position 1 of "2-DMAECE", "2-MorECE", "2-PyECE", and "2-BimECE", respectively.

[0033] "2-4 crosslinked sugar" refers to a sugar in which a crosslinking unit is formed by substitution at two positions, the 2nd and 4th positions, in ribose. Examples of crosslinking units include a C2-6 alkylene group (the alkylene group is either unsubstituted or substituted with one or more substituents selected from the group consisting of halogen atoms, oxo groups, and thioxo groups, and one or two methylene groups of the alkylene group are either not substituted or independently substituted with groups selected from the group consisting of -O-, -NR10- (where R10 represents a hydrogen atom, a C1-6 alkyl group, or a halo-C1-6 alkyl group), and -S-).

[0034] "2'-4'-bridged nucleoside" (2',4'-BNA) refers to a molecule having a nucleic acid base at position 1 of the aforementioned 2'-4'-bridged sugar. For example, LNA (Locked Nucleic Acid) described later. Also known as Acid(registered trademark), this includes β-D-methyleneoxy(4'-CH2-O-2')BNA or α-L-methyleneoxy(4'-CH2-O-2')BNA, ethyleneoxy(4'-(CH2)2-O-2')BNA, β-D-thio(4'-CH2-S-2')BNA, aminooxy(4'-CH2-ON(R11)-2')BNA (where R11 is H or CH3), oxyamino(4'-CH2-N(R12)-O-2')BNA (where R12 is H or CH3), 2',4'-BNACOC, and 3'-amino-2',4'-BNA. Examples include 5'-methyl BNA, (4'-CH(CH3)-O-2')BNA (also known as cEt), (4'-CH(CH2OCH3)-O-2')BNA (also known as cMOE-BNA), amide-type BNA (4'-C(=O)-N(R13)-2')BNA (R13 is H or CH3) (also known as AmNA), (4'-C(spirocyclopropyl)-O-2')BNA (also known as scpBNA), (4'-CH2-N(R14)-2')BNA (R14 is C(=NH2+)NHR15, and R15 is H or CH3), and other BNAs known to those skilled in the art.

[0035] In the aforementioned "deoxyribonucleosides," "ribonucleosides," "2'-modified nucleosides," and "2'-4'-bridged nucleosides," the bond between the carbon atom at the 1' position and the nucleic acid base can be an α-glycosidic bond or a β-glycosidic bond, but it is usually a β-glycosidic bond. Therefore, β-D-methyleneoxyBNA is usually used as the LNA.

[0036] "n-" stands for normal, "s-" for secondary, and "t-" for tertiary.

[0037] "Halogen atom" or "halo" refers to a fluorine atom, chlorine atom, bromine atom, or iodine atom.

[0038] "C1-6 alkyl group" refers to a linear or branched saturated hydrocarbon group having 1 to 6 carbon atoms, such as methyl group, ethyl group, n-propyl group, isopropyl group, n-butyl group, isobutyl group, s-butyl group, t-butyl group, n-pentyl group, isopentyl group, neopentyl group, n-hexyl group, and isohexyl group (including various isomers).

[0039] "C1-10 alkyl group" refers to a linear or branched alkyl group having 1 to 10 carbon atoms. Examples include, in addition to the "C1-6 alkyl group" examples above, heptyl group, octyl group, nonyl group, decyl group (including various isomers), etc.

[0040] "C5-32 alkyl group" refers to a linear or branched alkyl group having 5 to 32 carbon atoms, and examples include pentyl group, hexyl group, heptyl group, octyl group, nonyl group, decyl group, undecyl group, dodecyl group, tridecyl group, tetradecyl group, pentadecyl group, hexadecyl group, heptadecyl group, octadecyl group, nonadecyl group, icosyl group, docosyl group, tetracosyl group, hexacosyl group, octacosyl group, triacontyl group, hentriacontyl group, dotriacontyl group (including various isomers), etc.

[0041] "C8-28 alkyl group" refers to a linear or branched alkyl group having 8 to 28 carbon atoms, and examples include octyl group, nonyl group, decyl group, undecyl group, dodecyl group, tridecyl group, tetradecyl group, pentadecyl group, hexadecyl group, heptadecyl group, octadecyl group, nonadecyl group, eicosyl group, docosyl group, tetracosyl group, hexacosyl group, octacosyl group (including various isomers), etc.

[0042] "C10-20 alkyl group" refers to a linear or branched alkyl group having 10 to 20 carbon atoms, and examples include decyl group, undecyl group, dodecyl group, tridecyl group, tetradecyl group, pentadecyl group, hexadecyl group, heptadecyl group, octadecyl group, nonadecyl group, eicosyl group (including various isomers), etc. "C14 alkyl group" refers to a linear or branched alkyl group having 14 carbon atoms. Examples include 1-tetradecyl group, 7-tetradecyl group, 1-methyltridecyl group, and 12-methyltridecyl group.

[0043] "C5-32 alkenyl group" refers to a straight-chain or branched unsaturated hydrocarbon group having 5 to 32 carbon atoms and containing one or more carbon-carbon double bonds. Examples include pentenyl group, hexenyl group, heptenyl group, octenyl group, nonenyl group, decenyl group, undecenyl group, dodecenyl group, tridecenyl group, tetradecenyl group, pentadecenyl group, hexadecenyl group, heptadecenyl group, octadecenyl group, nonadecenyl group, icocenyl group, dococenyl group, tetracocenyl group, hexacocenyl group, octacocenyl group, triacontenyl group, hentriacontenyl group, dotriacontenyl group (including various isomers), etc. A "C14 alkenyl group" refers to a linear or branched alkenyl group with 14 carbon atoms. Examples include the tetradeca-2-enyl group, tetradeca-7-enyl group, 1-methyltrideca-2-enyl group, and 12-methyltrideca-2-enyl group.

[0044] "Halo C1-6 alkyl group" means a group in which a hydrogen atom at any position of the "C1-6 alkyl group" is substituted with one or more of the "halogen atoms".

[0045] A "C2-6 alkylene group" refers to a divalent group (alkanediyl group) obtained by removing two hydrogen atoms from arbitrary positions from a linear or branched saturated hydrocarbon group with 2 to 6 carbon atoms. Examples include ethylene (ethanediyl) group, propane-1,3-diyl (trimethylene) group, propane-2,2-diyl group, 2,2-dimethyl-propane-1,3-diyl group, hexane-1,6-diyl (hexamethylene) group, and 3-methylbutane-1,2-diyl group.

[0046] "C1-10 alkylene group" refers to a linear or branched alkylene group having 1 to 10 carbon atoms. Examples include, in addition to the "C2-6 alkylene group" examples above, methylene group, propylene group, tetramethylene group, 1-methylpropylene group, 2-methylpropylene group, dimethylethylene group, ethylethylene group, pentamethylene group, 1-methyltetramethylene group, 2-methyltetramethylene group, 1,1-dimethyl-trimethylene group, 1,2-dimethyl-trimethylene group, 1-ethyl-trimethylene group, octamethylene group, and decamethylene group.

[0047] "C2-8 alkylene group" refers to a linear or branched alkylene group having 2 to 8 carbon atoms. Examples include, in addition to the "C2-6 alkylene group" examples above, methylene group, propylene group, tetramethylene group, 1-methylpropylene group, 2-methylpropylene group, dimethylethylene group, ethylethylene group, pentamethylene group, 1-methyltetramethylene group, 2-methyltetramethylene group, 1,1-dimethyl-trimethylene group, 1,2-dimethyl-trimethylene group, 1-ethyl-trimethylene group, and octamethylene group.

[0048] "C3-6 alkylene group" refers to a linear or branched alkylene group having 3 to 6 carbon atoms. Examples include propylene group, tetramethylene group, 1-methylpropylene group, 2-methylpropylene group, dimethylethylene group, ethylethylene group, pentamethylene group, 1-methyltetramethylene group, 2-methyltetramethylene group, 1,1-dimethyl-trimethylene group, 1,2-dimethyl-trimethylene group, 1-ethyl-trimethylene group, and hexamethylene group.

[0049] A "C6 alkylene group" refers to a linear or branched alkylene group having six carbon atoms. Examples include the hexamethylene group, 1-methyl-pentamethylene group, 2-methyl-pentamethylene group, and 1,1-dimethyl-tetramethylene group.

[0050] A "C1-6 alkoxy group" refers to a group in which the aforementioned "C1-6 alkyl group" is bonded to an oxy group.

[0051] "Halo-C1-6 alkoxy group" means a group in which a hydrogen atom at any position of the "C1-6 alkoxy group" is substituted with one or more of the "halogen atoms".

[0052] A "mono- or di-C1-6 alkylamino group" refers to a group in which one hydrogen atom of an amino group is replaced by one "C1-6 alkyl group," or a group in which two hydrogen atoms of an amino group are replaced by two identical or different "C1-6 alkyl groups." Examples include methylamino group, ethylamino group, propylamino group, isopropylamino group, butylamino group, dimethylamino group, diethylamino group, dipropylamino group, dibutylamino group, and N-ethyl-N-methylamino group.

[0053] "3- to 6-membered rings" refers to monocyclic saturated or unsaturated hydrocarbon rings with 3 to 6 carbon atoms, such as cyclopropane, cyclobutane, and cyclohexane.

[0054] A "5-10 membered heterocyclic group" refers to a 5- to 10 membered monocyclic or fused polycyclic aromatic or aromatic heterocyclic group containing 1 to 4 heteroatoms selected from nitrogen, sulfur, and oxygen atoms in addition to carbon atoms as ring constituent atoms. Suitable examples of the aforementioned "5-10 membered heterocyclic group" include thienyl group, furyl group, pyrrolyl group, imidazolyl group, pyrazolyl group, thiazolyl group, isothiazolyl group, oxazolyl group, isoxazolyl group, pyridyl group, pyrazinyl group, pyrimidinyl group, pyridadinyl group, triazolyl group, tetrazolyl group, triazinyl group, benzothiophenyl group, benzofuranyl group, benzimidazolyl group, benzoxazolyl group, benzoisoxazolyl group, benzothiazolyl group, benzoisothiazolyl group, benzotriazolyl group, imidazopyridinyl group, thienopyridinyl group, pyrrolopyridinyl group, pyrazolopyridinyl group, oxazolopyridinyl group, thiazolopyridinyl group, imidazopyradinyl group, imidazopyridinyl group, aziridinyl group, Examples include oxyranyl group, azetidinyl group, oxetanyl group, thietanyl group, tetrahydrothienyl group, tetrahydrofuranyl group, pyrrolinyl group, pyrrolidinyl group, oxopyrrolidinyl group, imidazolinyl group, oxoimidazolinyl group, imidazolidinyl group, oxazolinyl group, oxazolidinyl group, pyrazolinyl group, pyrazolidinyl group, thiazolinyl group, thiazolidinyl group, tetrahydroisothiazolyl group, tetrahydrooxazolyl group, tetrahydroisoxazolyl group, piperidinyl group, piperazinyl group, tetrahydropyridinyl group, dihydropyridinyl group, tetrahydropyridazinyl group, dihydropyranyl group, tetrahydropyranyl group, tetrahydrothiopyranyl group, morpholinyl group, and thiomorpholinyl group.

[0055] An "oxo group" refers to a group in which an oxygen atom is substituted via a double bond (=O). When an oxo group is substituted for a carbon atom, it forms a carbonyl group together with that carbon atom. A "thioxo group" refers to a group (=S) in which a sulfur atom is substituted via a double bond. When a thioxo group is substituted on a carbon atom, it forms a thiocarbonyl group together with that carbon atom.

[0056] "Nucleic acid bases" are purine bases or pyrimidine bases, and may be naturally occurring nucleic acid bases or modified naturally occurring nucleic acid bases. Examples of naturally occurring nucleic acid bases include adenine (A), guanine (G), thymine (T), cytosine (C), and uracil (U). The aforementioned "nucleic acid bases" include naturally occurring nucleic acid bases and the "modified nucleic acid bases" described below.

[0057] Examples of modifications to nucleic acid bases in "modified nucleic acid bases" include halogenation, methylation, ethylation, n-propylation, isopropylation, cyclopropylation, n-butylation, isobutylation, s-butylation, t-butylation, cyclobutylation, hydroxylation, amination, thioation, and demethylation. More specifically, examples include 5-methylation, 5-fluoration, 5-bromination, 5-iodation, and N4-methylation of cytosine; 2-thioation, 5-demethylation, 5-fluoration, 5-bromination, and 5-iodation of thymine; 2-thioation, 5-fluoration, 5-bromination, and 5-iodation of uracil; N6-methylation and 8-bromination of adenine; and N2-methylation and 8-bromination of guanine. Furthermore, examples of modifications to the nucleic acid base portion of nucleosides are disclosed in Journal of Medicinal Chemistry, 2016, 59, pp 9645-9667, Medicinal Chemistry Communication, 2014, 5, pp 1454-1471, Future Medicinal Chemistry, 2011, 3, pp 339-365, and International Publication No. 2018 / 155450, among others.

[0058] The nucleic acid base in the nucleoside is preferably at least one selected from the group consisting of adenine, guanine, thymine, cytosine, uracil, and 5-methylcytosine.

[0059] "5-methylcytosine" refers to cytosine that has a methyl group at the 5th position.

[0060] "Nucleic acid base sequence" refers to the sequence of nucleic acid bases from the 5' end to the 3' end of each nucleoside contained in an oligonucleotide.

[0061] "Continuous nucleic acid bases" refers to the sequence of a portion of nucleic acid bases that are continuous in the aforementioned "nucleic acid base sequence," from the 5' end to the 3' end.

[0062] "Nucleoside-inter-nucleoside bond" refers to a group or bond that forms a covalent bond between adjacent nucleosides in an oligonucleotide. "Nucleoside-inter-nucleoside bonds" include phosphodiester bonds and "modified nucleoside-inter-nucleoside bonds" as described below.

[0063] "Modified nucleoside bonds" refer to modified phosphodiester bonds, such as phosphorothioate bonds, methylphosphonate bonds (including chiral-methylphosphonate bonds), methylthiophosphonate bonds, phosphorodithioate bonds, phosphoramidate bonds, phosphorodiamidate bonds, phosphoramidothioate bonds, and boranophosphate bonds. Examples of phosphodiester bond modifications are disclosed in Journal of Medicinal Chemistry, 2016, 59, pp 9645-9667, Medicinal Chemistry Communication, 2014, 5, pp 1454-1471, Future Medicinal Chemistry, 2011, 3, pp 339-365, etc., and can be used for modified phosphodiester bonds.

[0064] "Modified nucleoside" refers to a nucleoside that has a modified sugar and / or a modified nucleic acid base.

[0065] "Oligononucleotide" means a molecule having a structure in which two or more identical or different "nucleosides" are linked by "nucleoside bonds" (e.g., phosphodiester bonds or modified phosphodiester bonds) that are independently selected from each other. "Oligodeoxyribonucleotide" means a polynucleotide or oligonucleotide in which two or more identical or different "deoxyribonucleosides" are linked by "nucleoside bonds" that are independently selected from each other. "Oligoribonucleotide" means a polynucleotide or oligonucleotide in which two or more identical or different "ribonucleosides" are linked by "nucleoside bonds" that are independently selected from each other.

[0066] "DNA" means a polynucleotide or oligonucleotide in which two or more naturally occurring "deoxyribonucleosides" are linked together by phosphodiester bonds. The naturally occurring deoxyribonucleosides that make up DNA may be the same or different. "RNA" means a polynucleotide or oligonucleotide in which two or more naturally occurring "ribonucleosides" are linked together by phosphodiester bonds. The naturally occurring ribonucleosides that make up RNA may be the same or different.

[0067] An "oligonucleotide complex" refers to a complex formed by the hybridization of multiple oligonucleotides (molecules) that are not covalently linked to each other, resulting in the formation of a single complex.

[0068] "Antisense effect" refers to the control of the function of a target RNA by hybridizing a target RNA, selected in response to a target gene, with an oligonucleotide having a sequence complementary to its sub-sequence. For example, if the target RNA is mRNA, hybridization can inhibit the translation of the target RNA, alter splicing functions such as exon skipping, or degrade the target RNA due to recognition of the hybridized portion.

[0069] An "antisense oligonucleotide" (ASO) is an oligonucleotide that produces the aforementioned antisense effect. Examples include DNA, oligodeoxyribonucleotides, gapmer-type antisense oligonucleotides (also simply called "gapmers"), and mixed-mer-type antisense oligonucleotides (also simply called "mixmers"), but are not limited to these. RNA, oligoribonucleotides, or oligonucleotides designed to produce an antisense effect are also acceptable.

[0070] "Hybridization" refers to the act of forming a double helix with oligonucleotides or parts thereof that contain complementary sequences, and the phenomenon of oligonucleotides or parts thereof containing complementary sequences forming a double helix.

[0071] "Complementary" means that two nucleic acid bases can form a Watson-Crick base pair (native base pair) or a non-Watson-Crick base pair (Hoogsteen base pair, etc.) via hydrogen bonding. Two oligonucleotides or parts thereof can "hybridize" if their sequences are complementary. Two oligonucleotides or parts thereof do not need to be perfectly complementary to hybridize, but the complementarity required for two oligonucleotides or parts thereof to hybridize is preferably 70% or higher, more preferably 80% or higher, and even more preferably 90% or higher (e.g., 95%, 96%, 97%, 98%, or 99% or higher). Sequence complementarity is determined by using a computer program that automatically identifies subsequences of oligonucleotides. OligoAnalyzer is one such software, for example, provided by Integrated DNA Technologies. This program is also available on their website. When two oligonucleotides have the above-mentioned desirable complementarity, a person skilled in the art can determine that the two oligonucleotides hybridize.

[0072] "Gapmer" refers to an oligonucleotide that includes the "gap segment," "5' wing segment," and "3' wing segment," which will be described later.

[0073] The "gap segment" is a region containing "at least four consecutive nucleosides recognized by RNaseH," and is not particularly limited as long as it contains four or more consecutive nucleosides and is recognized by RNaseH, but the consecutive nucleosides are preferably selected independently from deoxyribonucleosides and sugar-modified nucleosides, and contain at least two deoxyribonucleosides. The nucleosides at the 5' and 3' ends of the gap segment are preferably deoxyribonucleosides.

[0074] The "5' wing segment" is connected to the 5' side of the gap segment and is a region containing "at least one nucleoside" but not "at least four consecutive nucleosides recognized by RNaseH". Here, the sugar portion of the nucleoside at the 3' end of the 5' wing segment is different from the sugar portion of the nucleoside at the 5' end of the gap segment. The boundary between the 5' wing segment and the gap segment is confirmed by the difference in the sugar portion. In one embodiment of the present invention, the difference in the sugar portion is preferably determined by the presence or absence of modification at the 2 position of the corresponding sugar. As defined above, a "2-modified sugar" is a non-crosslinked sugar in which the oxygen or carbon atom at the 2 position of ribose is modified, and a "2-4 crosslinked sugar" is a sugar in which the crosslinking unit is replaced by substitution at two positions, 2 and 4, of ribose as defined above; therefore, in both cases, the 2 position is modified. (For example, the nucleoside at the 5' end of the gap segment is a deoxyribonucleoside, and the nucleoside at the 3' end of the 5' wing segment is a sugar-modified nucleoside. This sugar-modified nucleoside is a 2'-4'-bridged nucleoside or a 2'-modified nucleoside.) The nucleoside at the 3' end of the 5' wing segment is generally a sugar-modified nucleoside. The 5' wing segment is not particularly limited as long as it satisfies the above definition, but the at least one nucleoside is preferably selected independently from a deoxyribonucleoside and a sugar-modified nucleoside, and contains at least one sugar-modified nucleoside.

[0075] The "3' wing segment" is connected to the 3' side of the gap segment and is a region containing "at least one nucleoside" but not "at least four consecutive nucleosides recognized by RNaseH". Here, the sugar moiety of the nucleoside at the 5' end of the 3' wing segment is different from the sugar moiety of the nucleoside at the 3' end of the gap segment. The boundary between the 3' wing segment and the gap segment is confirmed by the difference in the sugar moiety. In one embodiment of the present invention, the difference in the sugar moiety is preferably determined by the presence or absence of modification at the 2 position of the corresponding sugar. (For example, the nucleoside at the 3' end of the gap segment is a deoxyribonucleoside, and the nucleoside at the 5' end of the 3' wing segment is a sugar-modified nucleoside. This sugar-modified nucleoside is a 2'-4'-bridged nucleoside or a 2'-modified nucleoside.) The nucleoside at the 5' end of the 3' wing segment is generally a sugar-modified nucleoside. The 3' wing segment is not particularly limited as long as it satisfies the above definition, but the at least one nucleoside is preferably selected independently from a deoxyribonucleoside and a sugar-modified nucleoside, and contains at least one sugar-modified nucleoside.

[0076] One typical example is the 5'-wing segment, which is the portion where a 2'-modified nucleoside or a 2'-4'-bridged nucleoside is continuous from the 5' end, and the boundary between the 5'-wing segment and the gap segment is where the nucleoside at the 3' end of the 5'-wing segment connects to other nucleosides (such as deoxyribonucleosides and ribonucleosides) excluding the 2'-modified nucleoside or 2'-4'-bridged nucleoside. One typical example is the 3'-wing segment, where a 2'-modified nucleoside or a 2'-4'-bridged nucleoside is continuous from the 3' end, and the boundary between the 3'-wing segment and the gap segment is where the nucleoside at the 5' end of the 3'-wing segment connects to other nucleosides (such as deoxyribonucleosides and ribonucleosides) excluding the 2'-modified nucleoside or 2'-4'-bridged nucleoside.

[0077] RNaseH is generally known as a ribonuclease that recognizes double helix formed by the hybridization of DNA and RNA in living organisms, cleaving the RNA and producing single-stranded DNA. RNaseH can recognize not only double helix formed by the hybridization of DNA and RNA, but also double helix in which at least one of the nucleic acid base portion, phosphodiester bond portion, or sugar portion of at least one of the DNA or RNA is modified. For example, it can recognize double helix formed by the hybridization of phosphorothioate-modified DNA and RNA. It can also recognize double helix formed by the hybridization of oligodeoxyribonucleotides and oligoribonucleotides. Therefore, DNA can be recognized by RNaseH when hybridized with RNA. Similarly, RNA can be cleaved by RNaseH when hybridized with DNA. The same applies when at least one of the nucleic acid base portion, phosphodiester bond portion, and sugar portion of DNA or RNA is modified. For example, a typical example is an oligonucleotide in which the phosphodiester bond portion of DNA is modified with a phosphorothioate. Examples of DNA and / or RNA modifications that can be recognized by RNaseH are described, for example, in Nucleic Acids Research, 2014, 42, pp 5378-5389; Bioorganic & Medicinal Chemistry Letters, 2008, 18, pp 2296-2300; Molecular BioSystems, 2009, 5, pp 838-843; Nucleic Acid Therapeutics, 2015, 25, pp 266-274; and The Journal of Biological Chemistry, 2004, 279, pp 36317-36326. In the present invention, RNaseH is preferably mammalian RNaseH, more preferably human RNaseH, and particularly preferably human RNaseH1.

[0078] "At least four consecutive nucleosides recognized by RNaseH" includes four or more consecutive nucleosides and is not particularly limited as long as it is recognized by RNaseH, but examples include "at least four consecutive deoxyribonucleosides". The number of nucleosides constituting "at least four consecutive nucleosides recognized by RNaseH" is, for example, 5 to 30, preferably 5 to 15, more preferably 8 to 12, and particularly preferably 10. A person skilled in the art can determine whether a given sequence of at least four consecutive nucleosides is "at least four consecutive nucleosides recognized by RNaseH" by the structure of the sugar moiety of the sequence of nucleosides.

[0079] A mixmer is an oligonucleotide that contains multiple sugar-modified nucleosides and produces an antisense effect without having the gap segment. Examples include oligonucleotides in which nucleosides or oligonucleotides consisting of 1 to 20 sugar-modified nucleosides and nucleosides or oligonucleotides consisting of 1 to 3 deoxyribonucleosides are alternately linked, and oligonucleotides composed solely of sugar-modified nucleosides.

[0080] siRNA (small interfering RNA) is a small, double-stranded oligoribonucleotide. siRNA is involved in a phenomenon called RNA interference (RNAi), suppressing gene expression by disrupting target mRNA. siRNA typically consists of 15-30 base pairs, preferably 21-23 base pairs. The 3' ends of the two nucleic acid strands often have two protruding deoxythymidine molecules (commonly denoted as tt, or dTdT, etc.).

[0081] Aptamers are oligonucleotides that exhibit the ability to bind to specific extracellular proteins and inhibit their function.

[0082] A "ribozyme" is an RNA or oligoribonucleotide that possesses catalytic activity.

[0083] miRNA (micro-RNA) is RNA that is encoded in the genome but is not translated into protein (non-coding RNA) and has the effect of suppressing gene expression. It is typically an RNA molecule of 21-25 nucleotides.

[0084] "CpG oligonucleotides" are oligonucleotides containing deoxycytidine (C) and deoxyguanosine (G) that activate innate immunity by interacting with proteins such as Toll-like Receptor 9 (TLR9). Typically, they are oligodeoxyribonucleotides of 15 to 30 bases.

[0085] "Decoy nucleic acids" are double-stranded oligodeoxyribonucleotides that bind to transcription factors and suppress the transcription process. Decoy nucleic acids can suppress the gene expression of the transcription factor. Decoy nucleic acids typically consist of 15 to 30 base pairs.

[0086] <Lipid-binding oligonucleotides or their complexes> The present invention provides lipid-binding oligonucleotides or complexes thereof. The lipid-binding oligonucleotides or complexes thereof of the present invention have the following general formula (I): [ka] (In the formula, W is a group derived from an oligonucleotide compound or oligonucleotide complex. L is a divalent group derived from a substituted or unsubstituted C1-10 alkylene group or a substituted or unsubstituted polyC1-10 alkylene glycol. R1 and R2 may each be independently a substituted or unsubstituted C5-32 alkyl group or a substituted or unsubstituted C5-32 alkenyl group, or R1 and R2 may be bonded to each other to form a ring. It is represented as follows.

[0087] In the present invention, "substituted or unsubstituted C1-10 alkylene group" means an (unsubstituted) C1-10 alkylene group or a C1-10 alkylene group substituted with one or more arbitrary substituents.

[0088] In the present invention, "substituted or unsubstituted" means that the group is substituted with at least one substituent selected from a given group of substituents, such as halogen atoms, hydroxyl groups, carboxyl groups, C1-6 alkoxy groups, and aryl groups, or is unsubstituted.

[0089] "Substituted C1-10 alkylene group" means that one or more arbitrary hydrogen atoms of the alkylene group are substituted with substituents selected from the group consisting of halogen atoms, hydroxyl groups, carboxyl groups, C1-6 alkoxy groups, and aryl groups.

[0090] Examples of divalent groups derived from poly-C1-10 alkylene glycol include groups represented by the formula: -(O)l-(Alk-O-)m-Alk-(O)n- (wherein l and n are independently 0 or 1, m is an integer from 2 to 20, and Alk is a C1-10 alkylene group, which may be substituted by a group selected from the group consisting of a hydroxyl group, a protected hydroxyl group, a halogen atom, a hydroxyl group, a carboxyl group, a C1-6 alkoxy group, and an aryl group). The protected hydroxyl group is not particularly limited as long as it is stable when bonded to an oligonucleotide, and examples include ether-based protecting groups such as triarylmethyl (e.g., triphenylmethyl(trityl), monomethoxytrityl, dimethoxytrityl (DMTr), trimethoxytrityl, etc.); acetal-based protecting groups such as methoxymethyl, methylthiomethyl, methoxyethyl, benzyloxymethyl, 2-tetrahydropyranyl; acyl-based protecting groups such as acyl (e.g., formyl, acetyl, pivaloyl, benzoyl, etc.); and tri(alkyl)silyl (e.g., trimethylsilyl, triethylsilyl, Examples of silyl protecting groups include triisopropylsilyl, t-butyldimethylsilyl, dimethylisopropylsilyl, etc., (alkyl)diarylsilyl (e.g., t-butyldiphenylsilyl, diphenylmethylsilyl, etc.), triarylsilyl (e.g., triphenylsilyl, etc.), tribenzylsilyl, [(triisopropylsilyl)oxy]methyl (Tom group), etc.; 1-(4-chlorophenyl)-4-ethoxypiperidine-4-yl (Cpep group), 9-phenylxanthene-9-yl (Pixyl group), 9-(p-methoxyphenyl)xanthene-9-yl (MOX group), etc. The protecting group of the "protected hydroxyl group" is preferably benzoyl, trimethylsilyl, triethylsilyl, triisopropylsilyl, t-butyldimethylsilyl, triphenylmethyl, monomethoxytrityl, dimethoxytrityl, trimethoxytrityl, 9-phenylxanthene-9-yl, or 9-(p-methoxyphenyl)xanthene-9-yl.

[0091] Among these, the L in general formula (I) is preferably an unsubstituted C1-10 alkylene group, more preferably a C2-8 alkylene group, even more preferably a C3-6 alkylene group, and particularly preferably a C6 alkylene group. For example, ethylene, propylene, trimethylene, tetramethylene, hexamethylene, and octamethylene groups (including various isomers) are more preferred, trimethylene, tetramethylene, and hexamethylene groups (including various isomers) are even more preferred, and hexamethylene groups (including various isomers) are particularly preferred.

[0092] In the present invention, "substituted or unsubstituted C5-32 alkyl group" means an (unsubstituted) C5-32 alkyl group or a C5-32 alkyl group substituted with one or more arbitrary substituents.

[0093] "Substituted C5-32 alkyl group" means that one or more hydrogen atoms of the alkyl group are substituted with substituents selected from the group consisting of halogen atoms, hydroxyl groups, carboxyl groups, C1-6 alkoxy groups, and aryl groups.

[0094] Among these, R1 and R2 in general formula (I) are preferably unsubstituted C5-32 alkyl groups, more preferably C8-28 alkyl groups, even more preferably C10-20 alkyl groups, and particularly preferably C14 alkyl groups. For example, decyl groups, undecyl groups, dodecyl groups, tridecyl groups, tetradecyl groups, pentadecyl groups, hexadecyl groups, heptadecyl groups, octadecyl groups, nonadecyl groups, and eicosyl groups (including various isomers) are more preferred, and tetradecyl groups (including various isomers) are particularly preferred.

[0095] In the present invention, "substituted or unsubstituted C5-32 alkenyl group" means an (unsubstituted) C5-32 alkenyl group or a C5-32 alkenyl group substituted with one or more arbitrary substituents.

[0096] "Substituted C5-32 alkenyl group" means that one or more arbitrary hydrogen atoms of the alkenyl group are substituted with substituents selected from the group consisting of halogen atoms, hydroxyl groups, carboxyl groups, C1-6 alkoxy groups, and aryl groups.

[0097] Among these, R1 and R2 in general formula (I) are preferably unsubstituted C5-32 alkenyl groups, more preferably C8-28 alkenyl groups, even more preferably C10-20 alkenyl groups, and particularly preferably C14 alkenyl groups. For example, decenyl groups, undecenyl groups, dodecenyl groups, tridecenyl groups, tetradecenyl groups, pentadecenyl groups, hexadecenyl groups, heptadecenyl groups, octadecenyl groups, nonadecenyl groups, and eicocenyl groups (including various isomers) are even more preferred, and tetradecenyl groups (including various isomers) are particularly preferred.

[0098] In the present invention, "may be bonded to each other to form a ring" means that the substituted or unsubstituted C5-32 alkyl groups or substituted or unsubstituted C5-32 alkenyl groups of R1 and R2 may be bonded to each other in part, and including the "-O-CH2-CH-O-" portion in general formula (I) to which they are bonded, they may form a ring of 10 to 64 members. The 10 to 64-membered ring is preferably a 10 to 40-membered ring, more preferably a 12 to 30-membered ring, and even more preferably a 20 to 30-membered ring.

[0099] In the present invention, the "group derived from an oligonucleotide compound or oligonucleotide complex" means a substructure of an oligonucleotide compound formed by removing a hydrogen atom, a hydroxyl group, etc., from the 3' or 5' end of at least one oligonucleotide compound constituting the oligonucleotide compound or oligonucleotide complex. Therefore, one of the 3' or 5' ends of the oligonucleotide compound is covalently bonded to L in general formula (I). Preferably, one of the 3' or 5' ends of the oligonucleotide compound and L are linked by a phosphodiester bond or a modified phosphodiester bond, and more preferably by a phosphodiester bond.

[0100] The oligonucleotide compound of the present invention is not particularly limited as long as it is an oligonucleotide useful as a nucleic acid drug, and may be, for example, an antisense oligonucleotide, siRNA, aptamer, ribozyme, miRNA, CpG oligo, decoy nucleic acid, etc.

[0101] The oligonucleotide compound of the present invention is preferably an antisense oligonucleotide. Examples of antisense oligonucleotides include DNA, oligodeoxyribonucleotides, gapmer-type antisense oligonucleotides (or simply referred to as "gapmers"), and mixed-mer-type antisense oligonucleotides (or simply referred to as "mixmers"), but are not limited to these. RNA, oligoribonucleotides, or oligonucleotides designed to normally produce an antisense effect may also be used. Alternatively, the "oligonucleotide complex" of the present invention may include an HDO (hetero-double-stranded nucleic acid) containing an antisense oligonucleotide (first oligonucleotide) which is single-stranded DNA and a complementary RNA oligonucleotide (second oligonucleotide). Alternatively, the first and second oligonucleotides may be linked by a nucleic acid linker to form a single-stranded oligonucleotide compound, linked by a linking group containing a non-nucleotide structure, or directly linked to form a single-stranded oligonucleotide compound (single-stranded heteroduplex nucleic acid).

[0102] In the general formula (I) of the present invention, W is preferably a group derived from a gapmer-type antisense oligonucleotide. The group derived from a gapmer-type antisense oligonucleotide refers to a substructure formed by removing a hydrogen atom, a hydroxyl group, etc. from the 3' or 5' end of a gapmer, and consists of 10 to 40 nucleosides, preferably 13 to 25 nucleosides, and is covalently bonded to L at its 3' or 5' end and covalently bonded to L at its 5' end. Furthermore, the gapmer-type antisense oligonucleotide preferably contains at least four consecutive deoxyribonucleosides. Furthermore, the gapmer-type antisense oligonucleotide preferably includes at least one selected from the group consisting of 2′-modified non-crosslinked nucleosides and 2′-4′-crosslinked nucleosides.

[0103] In the general formula (I) of the present invention, W is preferably a group derived from a micmer-type antisense oligonucleotide. The group derived from a micmer-type antisense oligonucleotide refers to a substructure formed by removing a hydrogen atom, a hydroxyl group, etc., from the 3' or 5' end of a micmer, and consists of 10 to 40 nucleosides, preferably 13 to 25 nucleosides, and is covalently bonded to L at its 3' or 5' end and covalently bonded to L at its 5' end. Furthermore, the mixed-mer type antisense oligonucleotide preferably does not contain at least four consecutive deoxyribonucleosides. Furthermore, the mixed-mer type antisense oligonucleotide preferably includes at least one selected from the group consisting of 2′-modified non-crosslinked nucleosides and 2′-4′-crosslinked nucleosides.

[0104] Alternatively, W in the general formula (I) of the present invention is preferably an oligonucleotide complex, the group being derived from a double-stranded nucleic acid. Double-stranded nucleic acids are described, for example, as HDO (hetero-double-stranded nucleic acid) in International Publication No. 2013 / 089283, the entire publication of which is incorporated herein by reference. Preferably, the oligonucleotide complex is a double-stranded oligonucleotide complex comprising a first oligonucleotide and a second oligonucleotide, wherein the first oligonucleotide is a gapmer-type antisense oligonucleotide or a mixed-type antisense oligonucleotide consisting of 7 to 100 nucleosides, and the second oligonucleotide includes a sequence that enables hybridization with at least a portion of the first oligonucleotide and consists of 4 to 100 nucleosides independently selected from deoxyribonucleosides, ribonucleosides, and sugar-modified nucleosides, and the first oligonucleotide and the second oligonucleotide hybridize. The oligonucleotide complex is preferably covalently bonded to L via the 3' or 5' end of the first or second oligonucleotide, more preferably via the 3' or 5' end of the second oligonucleotide, and particularly preferably via the 5' end of the second oligonucleotide.

[0105] Alternatively, as W in the general formula (I) of the present invention, a group derived from a single-stranded oligonucleotide compound in which the first and second oligonucleotides of the oligonucleotide complex are linked by a nucleic acid linker is preferred. The nucleic acid linker is preferably a group derived from an oligonucleotide consisting of 2 to 10 nucleosides. Alternatively, as W in the general formula (I) of the present invention, a group derived from a single-chain oligonucleotide compound in which the first and second oligonucleotides of the oligonucleotide complex are linked by a linking group containing a non-nucleotide structure is preferred. Examples of linking groups containing a non-nucleotide structure include alkylene groups having 2 to 50 carbon atoms and groups derived from polyalkylene glycols. Alternatively, the W in the general formula (I) of the present invention is preferably a group derived from a single-stranded oligonucleotide compound in which the first and second oligonucleotides of the oligonucleotide complex are directly linked. The above aspects are described, for example, in International Publication Nos. 2017 / 131124, 2018 / 143475, and 2019 / 022196, which are incorporated herein by reference in their entirety.

[0106] The lipid-binding oligonucleotides, etc., according to this embodiment can be manufactured by the method described below. However, the manufacturing method described below is merely an example of a general manufacturing method and does not limit the manufacturing method of the oligonucleotides, etc., according to this embodiment.

[0107] [ka]

[0108] The symbols in the formula are the same as in the definition above, and "step" means process.

[0109] Compound A, the starting material for Step I, can be synthesized, for example, by the method described in Japanese Patent Publication No. 2018-532990 or Japanese Patent No. 6356171. Specifically, Compound A having diverse R1 and R2 can be synthesized from Compound A-1 described below by combining oxidation reactions known to those skilled in the art (for oxidation reactions, see, for example, Comprehensive Organic Transformations, Second Edition, by R.C. Larock, Wiley-VCH (1999), etc.).

[0110] [ka]

[0111] In the formula, R1 and R2 are the same as defined above. For example, to synthesize compound A-1 in which R1 and R2 are alkyl groups or alkenyl groups, the desired R1 can be introduced by alkylating or alkenylating the primary hydroxyl group of compound A-2 using an alkyl halide reagent corresponding to R1. Similarly, the desired R2 can be introduced by alkylating or alkenylating the secondary hydroxyl group using an alkyl halide reagent corresponding to R2. By deprotecting the P1-protected hydroxyl group of the resulting compound A-1 in which R1 and R2 are alkyl groups or alkenyl groups can be synthesized.

[0112] Compound A-1, in which R1 and R2 bond to each other to form a ring, can be obtained by introducing the desired R1 and R2 to the ends of R1 and R2 using an alkyl halide reagent having a double bond, and then performing an olefin metathesis reaction. Specific examples of olefin metathesis reactions include reacting a second-generation Grubbs catalyst, such as dichloro[1,3-bis(2,4,6-trimethylphenyl)-2-imidazolidinylidene](benzylidene)(tricyclohexylphosphine)ruthenium(II)) in a solvent. Furthermore, saturated ring compounds can be obtained by hydrogenating the olefin ring compound obtained by an olefin metathesis reaction with hydrogen and palladium carbon in a solvent. [ka]

[0113] In the formula, P1 and represent hydroxyl protecting groups, and the other symbols are as defined above.

[0114] (Step I) Amidation reaction with primary amine Compound B can be obtained by a condensation exchange reaction with a carboxylic acid (compound A) using a primary alkylamine to which L is bonded. For example, one method involves reacting compound A with 1 to 10 equivalents of the primary alkylamine in a solvent in the presence of 1-[bis(dimethylamino)methylene]-1H-1,2,3-triazolo[4,5-b]pyridinium 3-oxide hexafluorophosphate.

[0115] (Step II) Phosphytylation reaction Compound C can be obtained by phosphytylation of the hydroxyl group of compound B by a reaction known to those skilled in the art (for example, a reaction using disubstituted alkoxyphosphines). Specific examples of phosphytylation reactions include reacting 2-cyanoethyl N,N,N',N'-tetraisopropylphosphodiamidite in a solvent in the presence of 4,5-dicyanoimidazole.

[0116] (Step III) Bonding reaction between oligonucleotides and lipids Lipid-binding oligonucleotides can be synthesized using an automated nucleic acid synthesizer (e.g., M-8-SE (manufactured by Nippon Techno Co., Ltd.)) with compound C, commercially available nucleotides necessary for producing oligonucleotide compounds of the desired nucleotide sequence, and phosphoramidite reagents.

[0117] One embodiment of the present invention is shown in formula (II): [ka] (In the formula, L is a substituted or unsubstituted C1-10 alkylene group. R1 and R2 may each be independently a substituted or unsubstituted C5-32 alkyl group or a substituted or unsubstituted C5-32 alkenyl group, or R1 and R2 may be bonded to each other to form a ring. Each of the R3 groups may be independently a C1-10 alkyl group, or the two alkyl groups of R3 may be bonded to each other to form a ring. R4 is a 2-cyanoethyl group. It is a compound represented by [formula]. The compound of formula (II) is useful for producing lipid-binding oligonucleotides according to the present invention.

[0118] One embodiment of the present invention is a pharmaceutical composition comprising a lipid-binding oligonucleotide compound of general formula (I) or a complex thereof, and a pharmacologically acceptable carrier.

[0119] A pharmaceutical composition comprising the lipid-binding oligonucleotide of general formula (I) or a complex thereof according to the present invention can be formulated by known pharmaceutical methods. For example, it can be used enterally (oral, etc.) or non-enterally as a capsule, tablet, pill, liquid, powder, granule, fine granule, film coating agent, pellet, lozenge, sublingual, chewable, buccal, paste, syrup, suspension, elixir, emulsion, topical application, ointment, hard ointment, poultices, transdermal formulations, lotions, inhalants, aerosols, injections, suppositories, etc.

[0120] These formulations can be appropriately combined with carriers that are pharmacologically or food-grade, specifically sterile water or physiological saline, vegetable oil, solvents, bases, emulsifiers, suspending agents, surfactants, pH adjusters, stabilizers, flavoring agents, fragrances, excipients, vehicles, preservatives, binders, diluents, isotonic agents, analgesics, bulking agents, disintegrants, buffers, coating agents, lubricants, colorants, sweeteners, viscosity modifiers, flavoring and odor modifiers, solubilizers, or other additives.

[0121] There are no particular limitations on the administration method of the pharmaceutical composition containing the lipid-binding oligonucleotide of general formula (I) of the present invention or a complex thereof, but examples include enteral (oral, etc.) or non-enteral administration. More preferably, examples include intravenous administration, intra-arterial administration, intraperitoneal administration, subcutaneous administration, intradermal administration, intra-airway administration, rectal administration, intramuscular administration, intrathecal administration, intraventricular administration, nasal administration, intravitreous administration, etc., and administration by infusion.

[0122] The diseases that can be treated, prevented, or improved by nucleic acid pharmaceuticals utilizing a pharmaceutical composition containing the lipid-binding oligonucleotide of general formula (I) of the present invention or a complex thereof are not particularly limited, and include, for example, metabolic diseases, cardiovascular diseases, tumors, infectious diseases, eye diseases, inflammatory diseases, autoimmune diseases, rare genetic diseases, and diseases caused by gene expression. More specifically, hypercholesterolemia, hypertriglyceridemia, spinal muscular atrophy, muscular dystrophy (Duchenne muscular dystrophy, myotonic dystrophy, congenital muscular dystrophy (Fukuyama type congenital muscular dystrophy, Ullrich type congenital muscular dystrophy, merosin-deficient congenital muscular dystrophy, integrin deficiency, Walker-Warburg syndrome, etc.), Becker type muscular dystrophy, limb-girdle type muscular dystrophy, Miyoshi type muscular dystrophy, facioscapulohumeral type muscular dystrophy, etc.), Huntington's disease, and Examples include Luzheimer's disease, transthyretin amyloidosis, familial amyloid cardiomyopathy, multiple sclerosis, Crohn's disease, inflammatory bowel disease, acromegaly, type 2 diabetes, chronic nephropathy, RSV infection, Ebola hemorrhagic fever, Marburg fever, HIV, influenza, hepatitis B, hepatitis C, cirrhosis, chronic heart failure, myocardial fibrosis, atrial fibrillation, prostate cancer, melanoma, breast cancer, pancreatic cancer, colorectal cancer, renal cell carcinoma, cholangiocarcinoma, cervical cancer, liver cancer, lung cancer, leukemia, non-Hodgkin lymphoma, atopic dermatitis, glaucoma, and age-related macular degeneration. Depending on the type of disease, the gene causing the disease can be set as the target gene, and further, the expression control sequence (e.g., antisense sequence) can be appropriately set according to the sequence of the target gene.

[0123] In addition to primates such as humans, various diseases in other mammals can be treated, prevented, or improved by pharmaceutical compositions containing the lipid-binding oligonucleotide of general formula (I) of the present invention or a complex thereof. For example, but not limited to, diseases of mammalian species including cows, sheep, goats, horses, dogs, cats, guinea pigs, or other rodent species such as bovines, ovines, equines, canines, felines, and mice can be treated. Furthermore, compositions containing antisense oligonucleotides can also be applied to other species such as birds (e.g., chickens).

[0124] When a pharmaceutical composition containing the lipid-binding oligonucleotide of general formula (I) of the present invention or a complex thereof is administered to or ingested by an animal, including a human, the dosage or intake is appropriately selected according to the age, weight, symptoms, health condition, and type of composition (pharmaceutical, food, etc.) of the subject, but the dosage or intake is preferably 0.0001 mg / kg / day to 100 mg / kg / day in terms of lipid-binding oligonucleotide.

[0125] The lipid-binding oligonucleotide of general formula (I) or its complex according to the present invention is delivered to target organs more efficiently than conventional oligonucleotides, and therefore an enhancement of pharmacological effects can be expected. Accordingly, it is possible to provide a method for more safely controlling the expression of target genes by administering the lipid-binding oligonucleotide of general formula (I) or its complex according to the present invention to animals, including humans. Furthermore, it is also possible to provide a method for treating, preventing, and improving various diseases involving the control of target genes, which includes administering a composition containing the lipid-binding oligonucleotide of general formula (I) or its complex according to the present invention to mammals, including humans.

[0126] Preferred methods for using the lipid-binding oligonucleotide or complex thereof of the present invention, as described above, include the following. - A method for controlling the function of a target RNA, comprising the step of contacting a cell with a lipid-binding oligonucleotide of general formula (I) or a complex thereof according to the present invention. - A method for controlling the function of a target RNA in a mammal, comprising the step of administering a pharmaceutical composition containing a lipid-binding oligonucleotide of the general formula (I) of the present invention or a complex thereof to the mammal. - Use of the lipid-binding oligonucleotide of general formula (I) of the present invention or a complex thereof for controlling the function of a target RNA in mammals. - Use of the lipid-binding oligonucleotide of general formula (I) of the present invention or a complex thereof for producing a drug for controlling the function of a target RNA in mammals. - A method for controlling the expression of a target gene, comprising the step of contacting a cell with a lipid-binding oligonucleotide of the general formula (I) of the present invention or a complex thereof. - A method for controlling the expression of a target gene in a mammal, comprising the step of administering a pharmaceutical composition containing a lipid-binding oligonucleotide of the general formula (I) of the present invention or a complex thereof to the mammal. - Use of the lipid-binding oligonucleotide of general formula (I) of the present invention or a complex thereof for controlling the expression of a target gene in mammals. - Use of the lipid-binding oligonucleotide of general formula (I) of the present invention or a complex thereof for producing a drug for controlling the expression of a target gene in mammals.

[0127] In the present invention, the control of the function of the target RNA means, for example, that the function of the target RNA is suppressed by the degradation of the target RNA, which may occur when the antisense sequence portion coats a part of the target RNA by hybridization, thereby regulating or modifying the splicing function, such as inhibiting translation or exon skipping, or when the portion formed by the hybridization of the antisense sequence portion and a part of the target RNA is recognized.

[0128] The mammal is preferably a human. The route of administration is preferably enteral. In other embodiments, the route of administration is non-enteral. [Examples]

[0129] The present invention will be described in more detail below based on examples, but the following examples do not limit the scope of the present invention in any way. In the examples, "NMR" refers to nuclear magnetic resonance, and "(v / v)" refers to volume / volume. If 1H NMR data is provided, it is measured at 300 MHz (JNM-ECX300; JEOL Ltd., or JNM-ECP400; JEOL Ltd.), and represents the chemical shift δ (unit: ppm) of the signal (splitting pattern, integral value) with tetramethylsilane as the internal standard. "d" means doublet, "t" means triplet, "dd" means doublet of doublets, "m" means multiplet, "J" means coupling constant, and "CDCl3" means deuterated chloroform. Purification by silica gel column chromatography was performed using Purif-Pack(registered trademark)-EX (SI-50μm) manufactured by SHOKO SCIENCE.

[0130] [Manufacturing Example 1] [ka]

[0131] Under an argon atmosphere, a solution of compound 1 (10.0 g, 54.9 mmol) in dimethylformamide (100 mL) was mixed with sodium hydride (55 wt% liquid paraffin, 8.38 g, 192 mmol) and 1-bromotetradecane (67.1 mL, 247 mmol), and the mixture was heated and stirred overnight at 80°C (approximately 16-20 hours). After cooling to room temperature, water was added to the reaction mixture, and then it was concentrated. The residue was extracted with a mixed solvent of hexane and ethyl acetate, and then washed with dilute hydrochloric acid, water, and brine in that order. After concentrating the organic layer, the resulting crude compound 2 was dissolved in ethyl acetate (100 mL) and methanol (100 mL), 10% palladium-carbon (5.0 g) was added, and the mixture was stirred overnight under a hydrogen atmosphere (approximately 16-20 hours). The reaction mixture was filtered through Celite, and the filtrate was washed with a mixed solution of ethyl acetate, ethanol, and water. The filtrate and washing solution were then concentrated to obtain the crude product, which was purified by silica gel column chromatography (hexane / ethyl acetate) to obtain the target compound 3 (17.8 g, yield 67%) as a white solid. 1H NMR (CDCl3, 300 MHz) δ 0.88 (3H, t, 6.6 Hz), 1.19-1.38 (44H, m), 1.50-1.62 (4H, m), 2.16 (1H, t, 6.3Hz), 3.40-3.66 (8H, m), 3.68-3.77 (1H, m).

[0132] Compound 3 (17.4 g, 35.9 mmol) was mixed with methylene chloride (120 mL) and water (60 mL). 2-Azaadamantane-N-hydroxyl (AZADOL®) (457 mg, 1.08 mmol) and diacetoxyiodobenzene (34.7 g, 108 mmol) were added, and the mixture was stirred overnight at room temperature (approximately 16-20 hours). Subsequently, aqueous sodium thiosulfate solution was added to the reaction mixture, followed by extraction with methylene chloride. The organic layer was washed with water and dried with sodium sulfate. After concentration, methanol was added to the crude product, which was vigorously stirred and then filtered. This procedure was repeated three times to obtain Compound 4 (16.3 g, 91% yield) as a white solid. 1H NMR (CDCl3, 400 MHz) δ 0.88 (3H, t, 7.2 Hz), 1.20-1.37 (44H, m), 1.53-1.68 (4H, m), 3.43-3.53 (2H, m), 3.61-3.67 (2H, m), 3.71 (1H, dd, 10.6Hz, 5.0Hz), 3.80 (1H, dd, 10.6Hz, 3.2Hz), 4.04 (1H, dd, 5.0Hz, 3.2Hz).

[0133] Under an argon atmosphere, a solution of compound 4 (11.0 g, 22.1 mmol) in methylene chloride (100 mL) was mixed with triethylamine (4.59 mL, 66.4 mmol), 1-[bis(dimethylamino)methylene]-1H-1,2,3-triazolo[4,5-b]pyridinium 3-oxide hexafluorophosphate (HATU) (10.1 g, 26.6 mmol), and 6-amino-1-hexanol (3.11 g, 26.6 mmol), and the mixture was stirred overnight at room temperature (approximately 16-20 hours). Subsequently, water was added to the reaction mixture, followed by extraction with methylene chloride, and the organic layer was washed with dilute hydrochloric acid and brine. After concentration, ethyl acetate was added to the resulting crude product, which was vigorously stirred and then filtered. This procedure was repeated three times to obtain the target compound 5 (11.8 g, yield 87%) as a white solid. 1H NMR (CDCl3, 400 MHz) δ 0.88 (3H, t, 7.2Hz), 1.21-1.44 (48H, m), 1.49-1.64 (9H, m), 3.28 (1H, dd, 13.6Hz, 6.8Hz), 3.38-3.53 (3H, m), 3.38-3.53 (3H, m), 3.59-3.66 (4H, m), 3.77 (1H, dd, 10.6Hz, 2.8Hz), 3.87 (1H, dd, 5.4Hz, 2.4Hz), 6.70 (1H, t, 6.0Hz).

[0134] Compound 5 (4.58 g, 7.66 mmol) was concentrated with toluene. Under an argon atmosphere, 2-cyanoethyl N,N,N',N'-tetraisopropylphosphodiamidite (3.40 mL, 10.7 mmol) was added to a solution of the concentrated compound 5 in methylene chloride (40 mL), and the mixture was stirred at room temperature. Subsequently, a solution of 4,5-dicyanoimidazole (628 mg, 5.36 mmol) in acetonitrile was added, and the mixture was stirred for 2 hours. After concentration, methylene chloride was added, and the mixture was filtered to remove the white precipitate. The filtrate was concentrated and purified by silica gel column chromatography (hexane / ethyl acetate / triethylamine) to obtain the target compound 6 (3.22 g, 53%) as a white solid. 1H NMR (CDCl3, 300 MHz) δ 0.88 (3H, t, 6.6Hz), 1.17 (3H, d, 3.9Hz), 1.19 (3H, d, 3.6Hz), 1.23-1.38 (48H, m), 1.47-1.66 (8H, m), 2.64 (1H, t, 6.9Hz), 3.26 (1H, dd, 13.7Hz, 5.7Hz), 3.36-3.68 (9H,m), 3.74-3.89 (4H, m), 6.68 (1H, t, 6.3Hz).

[0135] [Manufacturing Example 2] [ka]

[0136] Under an argon atmosphere, a solution of compound 1 (3.00 g, 16.5 mmol) in dimethylformamide (30 mL) was mixed with sodium hydride (55 wt% liquid paraffin, 2.51 g, 57.6 mmol) and 1-bromohexadecane (22.6 mL, 74.1 mmol), and the mixture was heated and stirred overnight at 80°C (approximately 16-20 hours). After cooling to room temperature, water was added to the reaction mixture, and then it was concentrated. The residue was extracted with a mixed solvent of hexane and ethyl acetate, and then washed with dilute hydrochloric acid, water, and brine in that order. After concentrating the organic layer, the resulting crude compound 7 was dissolved in ethyl acetate (30 mL) and methanol (30 mL), 10% palladium-carbon (1.5 g) was added, and the mixture was stirred overnight under a hydrogen atmosphere (approximately 16-20 hours). The reaction mixture was filtered through Celite, and the filtrate was washed with a mixed solution of ethyl acetate, ethanol, and water. The filtrate and washings were concentrated to obtain the crude product, to which ethyl acetate and ethanol were added, and the mixture was vigorously stirred before filtration. This procedure was repeated three times to obtain the target compound 8 (7.79 g, 87% yield) as a white solid. 1H NMR (CDCl3, 300 MHz) δ 0.88 (6H, t, 6.5 Hz), 1.20-1.36 (52H, m), 1.50-1.62 (4H, m), 2.16 (1H, dd, 6.8Hz, 5.8Hz), 3.39-3.66 (8H, m), 3.68-3.77 (1H, m).

[0137] Starting with compound 8 (3.87 g, 7.15 mmol), compound 9 (2.68 g, 68% yield) was obtained as a white solid using the same method as in the synthesis of compound 4. 1H NMR (CDCl3, 400 MHz) δ 0.88 (6H, t, 6.2 Hz), 1.19-1.38 (52H, m), 1.52-1.68 (4H, m), 3.42-3.54 (2H, m), 3.59-3.73 (3H, m), 3.80 (1H, dd, 10.6Hz, 3.3Hz), 4.04 (1H, dd, 5.1Hz, 3.2Hz).

[0138] Starting with compound 9 (1.60 g, 2.88 mmol), compound 10 (1.61 g, yield 85%) was obtained as a white solid using the same method as in the synthesis of compound 5. 1H NMR (CDCl3, 400 MHz) δ 0.88 (6H, t, 7.0Hz), 1.19-1.44 (56H, m), 1.48-1.66 (9H, m), 3.28 (2H, dd, 13.9Hz, 7.0Hz), 3.38-3.54 (3H, m), 3.59-3.66 (4H, m), 3.77 (1H, dd, 10.6Hz, 2.8Hz), 3.87 (1H, dd, 5.1Hz, 2.2Hz), 6.70 (1H, t, 6.2Hz).

[0139] Compound 11 (1.93 g, 92% yield) was obtained as a white solid using compound 10 (1.60 g, 2.45 mmol) as the starting material, by the same method as the synthesis of compound 6. 1H NMR (CDCl3, 400 MHz) δ 0.88 (6H, t, 6.6Hz), 1.11-1.42 (68H, m), 1.47-1.66 (8H, m), 2.64 (2H, t, 6.6Hz), 3.18-3.32 (2H, m), 3.36-3.70 (9H, m), 3.71-3.90 (4H, m), 6.78 (1H, t, 5.9Hz).

[0140] [Manufacturing Example 3] [ka]

[0141] Under an argon atmosphere, a solution of compound 1 (3.00 g, 16.5 mmol) in dimethylformamide (30 mL) was mixed with sodium hydride (55 wt% liquid paraffin, 2.51 g, 57.6 mmol) and 1-bromododecane (17.8 mL, 74.1 mmol), and the mixture was heated and stirred overnight at 80°C (approximately 16-20 hours). After cooling to room temperature, water was added to the reaction mixture, and then it was concentrated. The residue was extracted with a mixed solvent of hexane and ethyl acetate, and then washed with dilute hydrochloric acid, water, and brine in that order. After concentrating the organic layer, the resulting crude compound 12 was dissolved in ethyl acetate (30 mL) and methanol (30 mL), 10% palladium-carbon (1.5 g) was added, and the mixture was stirred overnight under a hydrogen atmosphere (approximately 16-20 hours). The reaction mixture was filtered through Celite, and the filtrate was washed with a mixed solution of ethyl acetate, ethanol, and water. The filtrate and washing solution were then concentrated to obtain the crude product, which was purified by silica gel column chromatography (hexane / ethyl acetate) to obtain the target compound 13 (4.62 g, yield 65%) as a white solid. 1H NMR (CDCl3, 400 MHz) δ 0.88 (6H, t, 6.6 Hz), 1.20-1.37 (36H, m), 1.52-1.61 (4H, m), 2.19 (1H, t, 5.5Hz), 3.41-3.65 (8H, m), 3.69-3.76 (1H, m).

[0142] Starting with compound 13 (4.00 g, 9.33 mmol), crude compound 14 was obtained using the same method as in the synthesis of compound 4. Methanol was added to the obtained crude compound, and after vigorous stirring, the mixture was filtered to obtain compound 14 (2.00 g, yield 48%) as a white solid. 1H NMR (CDCl3, 400 MHz) δ 0.88 (6H, t, 6.6 Hz), 1.20-1.38 (36H, m), 1.51-1.68 (4H, m), 3.42-3.54 (2H, m), 3.64 (2H, t, 6.6Hz), 3.71 (1H, dd, 10.6Hz, 5.1Hz), 3.80 (1H, dd, 10.6Hz, 3.3Hz), 4.05 (1H, dd, 5.1Hz, 3.3Hz).

[0143] Using compound 14 (2.00 g, 4.51 mmol) as the starting material, crude compound 15 was obtained using the same method as in the synthesis of compound 5. Ethyl acetate was added to the obtained crude compound and vigorously stirred, then filtered to obtain compound 15 (1.23 g, yield 50%) as a white solid. 1H NMR (CDCl3, 400 MHz) δ 0.88 (6H, t, 6.6Hz), 1.21-1.44 (40H, m), 1.48-1.65 (9H, m), 3.28 (2H, dd, 12.8Hz, 6.6Hz), 3.38-3.54 (3H, m), 3.59-3.66 (4H, m), 3.77 (1H, dd, 10.6Hz, 2.6Hz), 3.87 (1H, dd, 5.1Hz, 2.6Hz), 6.70 (1H, t, 6.2Hz).

[0144] Compound 15 (1.20 g, 2.21 mmol) was used as the starting material, and compound 16 (1.21 g, yield 74%) was obtained as a white solid using the same method as in the synthesis of compound 6. 1H NMR (CDCl3, 400 MHz) δ 0.88 (6H, t, 6.6Hz), 1.17 (3H, d, 5.5Hz), 1.19 (3H, d, 5.1Hz), 1.22-1.43 (40H, m), 1.48-1.65 (8H, m), 2.64 (2H, t, 7.0Hz), 3.22-3.30 (2H, m), 3.38-3.68 (9H,m), 3.74-3.89 (4H, m), 6.69 (1H, t, 5.9Hz).

[0145] [Manufacturing Example 4] [ka]

[0146] Under an argon atmosphere, a solution of compound 1 (3.50 g, 19.2 mmol) in dimethylformamide (30 mL) was mixed with sodium hydride (55 wt% liquid paraffin, 2.51 g, 57.6 mmol) and 1-bromodecane (15.9 mL, 76.8 mmol), and the mixture was heated and stirred overnight at 80°C (approximately 16-20 hours). After cooling to room temperature, water was added to the reaction mixture, and then it was concentrated. The residue was extracted with a mixed solvent of hexane and ethyl acetate, and then washed with dilute hydrochloric acid, water, and brine in that order. After concentrating the organic layer, the resulting crude compound 17 was dissolved in ethyl acetate (30 mL) and methanol (30 mL), 10% palladium-carbon (1.5 g) was added, and the mixture was stirred overnight under a hydrogen atmosphere (approximately 16-20 hours). The reaction mixture was filtered through Celite, and the filtrate was washed with a mixed solution of ethyl acetate and methanol. The filtrate and washing solution were then concentrated to obtain a crude product, which was purified by silica gel column chromatography (hexane / ethyl acetate) to obtain the target compound 18 (6.08 g, yield 85%) as a white solid. 1H NMR (CDCl3, 400 MHz) δ 0.88 (6H, t, 6.6 Hz), 1.22-1.36 (28H, m), 1.52-1.62 (4H, m), 2.19 (1H, t, 6.6Hz), 3.41-3.66 (8H, m), 3.70-3.76 (1H, m).

[0147] Compound 18 (4.00 g, 10.7 mmol) was mixed with methylene chloride (40 mL) and water (20 mL). 2-Azaadamantane-N-hydroxyl (AZADOL®) (956 mg, 2.25 mmol) and diacetoxyiodobenzene (7.26 g, 22.5 mmol) were added, and the mixture was stirred overnight at room temperature (approximately 16-20 hours). Subsequently, an aqueous sodium thiosulfate solution was added to the reaction mixture, followed by extraction with methylene chloride. The organic layer was washed with water and dried with sodium sulfate. After concentration, methylene chloride (40 mL), triethylamine (1.66 mL, 12.0 mmol), 1-[bis(dimethylamino)methylene]-1H-1,2,3-triazolo[4,5-b]pyridinium 3-oxide hexafluorophosphate (HATU) (4.90 g, 12.9 mmol), and 6-amino-1-hexanol (1.51 g, 12.9 mmol) were added to the resulting compound 19, and the mixture was stirred overnight at room temperature (approximately 16-20 hours). Subsequently, water was added to the reaction mixture, and the mixture was extracted with methylene chloride. The organic layer was washed with dilute hydrochloric acid and brine. After concentration, the resulting crude product was purified by silica gel column chromatography (hexane / ethyl acetate) to obtain the target compound 20 (4.64 g, yield 89%) as a white solid. 1H NMR (CDCl3, 400 MHz) δ 0.86-0.91 (6H, m), 1.20-1.45 (32H, m), 1.48-1.65 (9H, m), 3.28 (2H, dd, 13.6Hz, 6.2Hz), 3.38-3.54 (3H, m), 3.59-3.66 (4H, m), 3.77 (1H, dd, 10.6Hz, 2.6Hz), 3.87 (1H, dd, 5.1Hz, 2.6Hz), 6.71 (1H, t, 5.9Hz).

[0148] Compound 21 (1.11 g, 52% yield) was obtained as a white solid using compound 20 (1.50 g, 2.29 mmol) as the starting material, by the same method as the synthesis of compound 6. 1H NMR (CDCl3, 400 MHz) δ 0.85-0.91 (6H, m), 1.17 (3H, d, 5.1Hz), 1.19 (3H, d, 5.5Hz), 1.21-1.41 (32H, m), 1.48-1.66 (8H, m), 2.65 (2H, t, 6.6Hz), 3.23-3.31 (2H, m), 3.38-3.69 (9H,m), 3.74-3.90 (4H, m), 6.69 (1H, t, 5.9Hz).

[0149] [Manufacturing Example 5] [ka]

[0150] Under an argon atmosphere, a solution of compound 1 (4.00 g, 22.0 mmol) in dimethylformamide (40 mL) was mixed with sodium hydride (55 wt% liquid paraffin, 2.87 g, 65.9 mmol) and 10-bromo-1-decene (17.7 mL, 87.8 mmol), and the mixture was heated and stirred overnight at 80°C (approximately 16-20 hours). After cooling to room temperature, water was added to the reaction mixture, and then it was concentrated. The residue was extracted with a mixed solvent of hexane and ethyl acetate, and then washed with water and then saline solution. After concentrating the organic layer, the resulting crude product was purified by silica gel column chromatography (hexane / ethyl acetate) to obtain the target compound 22 (6.16 g, yield 61%) as a colorless oil. 1H NMR (CDCl3, 400 MHz) δ 1.23-1.41 (20H, m), 1.50-1.61 (4H, m), 2.00-2.08 (4H, m), 3.43 (2H, t, 6.6Hz), 3.46-3.63 (7H, m), 4.55 (2H,s), 4.90-4.96 (2H, m), 4.95-5.03 (2H, m), 5.75-5.87 (2H, m), 7.27-7.36 (5H, m).

[0151] Under an argon atmosphere, (1,3-bis-(2,4,6-trimethylphenyl)-2-imidazolidinylidene)dichloro(ortho-isopropoxyphenylmethylene)ruthenium (Hoveyda-Grubbs catalyst®) (444 mg, 0.523 mmol) was added to a solution of compound 22 (4.80 g, 10.5 mmol) in 1,2-dichloromethane (1.05 L), and the mixture was stirred overnight at 80°C (approximately 16-20 hours). After the reaction was complete, the reaction mixture was allowed to cool to room temperature and concentrated. The resulting residue was filtered through silica gel and washed with a mixture of ethyl acetate and hexane. The concentrated crude compound 23 was dissolved in ethyl acetate (43 mL) and methanol (43 mL), 10% palladium-carbon (2.4 g) was added, and the mixture was stirred overnight under a hydrogen atmosphere (approximately 16-20 hours). The reaction mixture was filtered through Celite, and the filtrate was washed with a mixed solution of ethyl acetate and methanol. The filtrate and washing solution were then concentrated to obtain the crude product, which was purified by silica gel column chromatography (hexane / ethyl acetate) to obtain the target compound 24 (2.32 g, yield 65%) as a colorless oil. 1H NMR (CDCl3, 300 MHz) δ 1.23-1.44 (28H, m), 1.48-1.70 (4H, m), 2.14 (1H, s), 3.38-3.73 (9H, m).

[0152] Compound 26 (975 mg, 49% yield) was obtained as a colorless oily substance using compound 24 (1.50 g, 4.38 mmol) as the starting material, by the same method as the synthesis of compound 20. 1H NMR (CDCl3, 400 MHz) δ 1.29-1.50 (32H, m), 1.53-1.71 (8H, m), 3.32 (1H, dd, 13.9Hz, 7.0Hz), 3.47-3.55 (9H, m), 3.59-3.66 (9H, m), 3.68 (1H, t, 6.6Hz), 3.80-3.87 (1H, m), 3.95 (1H, dd, 6.2Hz, 2.2Hz), 6.71-6.81 (1H, m).

[0153] Compound 26 (970 mg, 2.13 mmol) was used as the starting material, and compound 27 (980 mg, 70% yield) was obtained as a colorless oil by the same method as the synthesis of compound 6. 1H NMR (CDCl3, 400 MHz) δ 1.17 (3H, d, 4.8Hz), 1.19 (3H, d, 5.1Hz), 1.27-1.43 (32H, m), 1.48-1.66 (8H, m), 2.65 (2H, t, 6.6Hz), 3.19-3.32 (2H, m), 3.41-3.51 (2H, m), 3.52-3.70 (7H, m), 3.75-3.93 (4H, m), 6.66-6.75 (1H, m).

[0154] In Table (1) of the Examples, "Compad No." refers to the compound number, and "Chemical Structure" refers to the chemical structure of each compound. The target gene is mouse Metastasis Associated in Lung Adenocarcinoma Transcript-1 (Malat1) or mouse Hypoxanthine Phosphoribosyltransferase 1 (Hprt1). In the sequence notation in the examples (Table 1), unless otherwise specified, "(L)" represents LNA (β-D-methyleneoxyBNA), lowercase letters represent deoxyribonucleosides, uppercase letters (excluding those with (L) above and (I) below) represent ribonucleosides, "^" represents a phosphorothioate bond, "5(x)" means that the nucleic acid base of the deoxyribonucleoside is 5-methylcytosine, and "5" in "5(L)" means that the nucleic acid base of the nucleoside is 5-methylcytosine. "I" means that the oxygen atom of the hydroxyl group at the 5' end is connected to the following phosphodiester bond via the formula (III) [ka] (In the formula, * represents the bond position with oligonucleotide Y. (* is the oxygen atom of the phosphodiester bond)) This means that a group represented by the following formula is bonded. "C16-" means that the oxygen atom of the hydroxyl group at the 5' end is bonded to the following oxygen atom via a phosphodiester bond. [ka] (In the formula, * represents the binding position with oligonucleotide Y. (* is the oxygen atom of the phosphodiester bond)) This means that the group represented by is bonded.

[0155] [Manufacturing Example 6] The antisense oligonucleotides (compounds represented by chemical structures corresponding to compound numbers) listed in Table 1 were prepared using the nucleic acid automated synthesizers nS-8II (Gene Design Co., Ltd.) and M-8-SE (Nippon Techno Service Co., Ltd.). Furthermore, intramolecular hybridization of the single-stranded oligonucleotides P22790171 and P22790146 was confirmed by non-denaturing polyacridamide gel electrophoresis. In non-denaturing polyacridamide gel electrophoresis, single-stranded DNA size markers (Gene Design Co., Ltd.) containing single-stranded DNA with nucleotide counts of 15, 20, 30, 40, 50, 60, and 80, and double-stranded RNA size markers (Gene Design Co., Ltd.) with base pair counts of 17, 21, 25, and 29 were used as markers, and a single band was confirmed for each single-stranded oligonucleotide.

[0156] [Table 1]

[0157] [Evaluation Example 1] Antisense suppression of mouse Malat1 in 3T3-L1 cells 3T3-L1 cells were seeded at a density of 20,000 cells / well in a 96-well plate. Approximately 24 hours later, the culture supernatant was replaced with D-MEM containing 2% FBS, and P21790027 and P22790171 were added to achieve a final concentration of 1000 nM (Free-Uptake). After 72 hours, cell lysates containing RNA were prepared using CellAmp® Direct RNA Prep Kit for RT-PCR (Real Time) (Takara Bio). The mouse Malat1 gene expression level was then measured by quantitative real-time PCR using TaqMan® Gene Expression Assays (Thermo Fisher Scientific) with One Step PrimeScript® III RT-qPCR Mix (Takara Bio). In real-time PCR, the mRNA amount of the housekeeping gene Gapdh was also quantified simultaneously, and the amount of Malat1 mRNA relative to the amount of Gapdh mRNA was evaluated as the Malat1 expression level. The results are shown in Table 2 as percentage expression of Malat1 compared to untreated control cells.

[0158] [Table 2]

[0159] [Evaluation Example 2] Antisense suppression of mouse Malat1 in N1E-115 cells N1E-115 cells were seeded at a density of 20,000 cells / well in a 96-well plate. Approximately 24 hours later, the culture supernatant was replaced with D-MEM without FBS, and P21790027 and P22790171 were added to achieve a final concentration of 1000 nM (Free-Uptake). After 24 hours, cell lysates containing RNA were prepared using CellAmp® Direct RNA Prep Kit for RT-PCR (Real Time) (Takara Bio). The mouse Malat1 gene expression level was then measured by quantitative real-time PCR using TaqMan® Gene Expression Assays (Thermo Fisher Scientific) with One Step PrimeScript® III RT-qPCR Mix (Takara Bio). In real-time PCR, the mRNA amount of the housekeeping gene Gapdh was also quantified simultaneously, and the amount of Malat1 mRNA relative to the amount of Gapdh mRNA was evaluated as the Malat1 expression level. The results are shown in Table 3 as percentage expression of Malat1 compared to untreated control cells.

[0160] [Table 3]

[0161] [Evaluation Example 3] Antisense suppression of Malat1 in mice C57BL / 6J mice (male, 6 weeks old, Jackson Laboratory Japan Co., Ltd.) were intravenously administered P21790027, P22790171, and P22790172 dissolved in physiological saline (Otsuka Saline Injection, Otsuka Pharmaceutical Co., Ltd.) at a dose equivalent to 0.95 μmol / kg of antisense oligonucleotide per mouse. As a control, only physiological saline (Otsuka Saline Injection, Otsuka Pharmaceutical Co., Ltd.) was administered. Heart, muscle, lung, liver, and kidney tissue was collected under isoflurane anesthesia 3 and 10 days after administration (P22790171 was collected only 3 days after administration). RNA was isolated from each organ using the MagMAX® mirVana® Total RNA Isolation Kit (Thermo Fisher Scientific). Then, the expression level of the mouse Malat1 gene was measured by quantitative real-time PCR using TaqMan® Gene Expression Assays (Thermo Fisher Scientific) with One Step PrimeScript® III RT-qPCR Mix (Takara Bio). The real-time PCR also quantified the mRNA amount of the housekeeping gene Gapdh, and the ratio of Malat1 mRNA to Gapdh mRNA was evaluated as the Malat1 expression level. The results, expressed as percentage expression of Malat1 relative to the untreated control group (control), are shown in Figure 1 (Day 10) and Figure 2 (Day 3).

[0162] As is clear from Figures 1 and 2, P22790171 and P22790172 showed a higher inhibitory effect on Malat1 expression in the heart, muscle, lung, and liver compared to P21790027.

[0163] [Evaluation Example 4] Antisense suppression of HPrt1 in mice C57BL / 6J mice (male, 6 weeks old, Jackson Laboratory Japan Co., Ltd.) were administered P22790163, P23790243, P23790244, and P23790246 dissolved in physiological saline (Otsuka Saline Injection, Otsuka Pharmaceutical Co., Ltd.) at a dose per mouse equivalent to 0.95 μmol / kg (P23790243). The drugs were administered intravenously at concentrations of 2.85 μmol / kg (P22790163, P23790243, and P23790244), 9.50 μmol / kg (P22790163, P23790243, and P23790244), and 19.5 μmol / kg (P22790163 only). As a control, only physiological saline (Otsuka Saline Injection, Otsuka Pharmaceutical Co., Ltd.) was administered. Tissue samples were collected from the heart (P22790163 and P23790244), liver (P22790163, P23790243, P23790244 and P23790246), lungs (P22790163, P23790243 and P23790244), and muscles (P22790163, P23790244 and P23790446) under isoflurane anesthesia 3, 10, and 20 days after administration (P23790246 was collected only 10 days after administration). RNA was isolated from each organ using the MagMAX® mirVana® Total RNA Isolation Kit (Thermo Fisher Scientific). Then, the mouse HPrt1 gene expression level was measured by quantitative real-time PCR using TaqMan® Gene Expression Assays (Thermo Fisher Scientific) with One Step PrimeScript® III RT-qPCR Mix (Takara Bio). In real-time PCR, the amount of mRNA of the housekeeping gene Gapdh was also quantified simultaneously, and the amount of HPrt mRNA relative to the amount of Gapdh mRNA was evaluated as the HPrt1 expression level.The results are shown as percentage expression of HPrt1 compared to the untreated control group (control) in Figures 3 (2.9 μmol / kg administration, liver), 4 (Day 10, liver), 5 (2.9 μmol / kg administration, heart), 6 (Day 10, heart), 7 (2.9 μmol / kg administration, lung), 8 (Day 10, muscle), and 9 (2.9 μmol / kg administration, muscle).

[0164] As is clear from Figures 3-9, P23790244 and P23790246 showed a higher inhibitory effect on HPrt1 expression in the liver, heart, lungs, and muscles compared to P22790163 and P23790243. [Industrial applicability]

[0165] The lipid-binding oligonucleotides of the present invention have been shown to enhance pharmacological effects not only in the liver but also in organs other than the liver, such as the heart, muscles, and lungs. Therefore, the lipid portion of the present invention is expected to be a means of delivering various oligonucleotides.

Claims

1. A lipid-binding oligonucleotide or its complex, represented by the following general formula (I). 【Chemistry 1】 (In the formula, W is a group derived from an oligonucleotide compound or oligonucleotide complex. L is a divalent group derived from a substituted or unsubstituted C1-10 alkylene group or a substituted or unsubstituted polyC1-10 alkylene glycol. R1 and R2 may each be independently a substituted or unsubstituted C5-32 alkyl group or a substituted or unsubstituted C5-32 alkenyl group, or R1 and R2 may be bonded to each other to form a ring.

2. The lipid-binding oligonucleotide or complex thereof according to claim 1, wherein R1 and R2 are each independently an unsubstituted C5-32 alkyl group or an unsubstituted C5-32 alkenyl group, or R1 and R2 may be bonded to each other to form a ring.

3. The lipid-binding oligonucleotide or complex thereof according to claim 1 or 2, wherein R1 and R2 are each independently an unsubstituted C5-32 alkyl group or an unsubstituted C5-32 alkenyl group.

4. The lipid-binding oligonucleotide or complex thereof according to any one of claims 1 to 3, wherein R1 and R2 are each independently an unsubstituted C10-20 alkyl group.

5. The lipid-binding oligonucleotide or complex thereof according to any one of claims 1 to 4, wherein R1 and R2 are unsubstituted C14 alkyl groups.

6. The lipid-binding oligonucleotide or complex thereof according to any one of claims 1 to 5, wherein L is an unsubstituted C1-10 alkylene group.

7. The lipid-binding oligonucleotide or complex thereof according to any one of claims 1 to 6, wherein L is an unsubstituted C6 alkylene group.

8. The lipid-binding oligonucleotide according to any one of claims 1 to 7, wherein the oligonucleotide compound is a gapmer-type antisense oligonucleotide.

9. The lipid-binding oligonucleotide according to claim 8, wherein the gapmer-type antisense oligonucleotide comprises at least one selected from the group consisting of 2'-modified non-crosslinked nucleosides and 2'-4'-crosslinked nucleosides.

10. The lipid-binding oligonucleotide according to claim 8 or 9, wherein the gapmer-type antisense oligonucleotide comprises at least four consecutive deoxyribonucleosides.

11. The lipid-binding oligonucleotide according to any one of claims 8 to 10, wherein the gapmer-type antisense oligonucleotide consists of 13 to 25 nucleosides.

12. The lipid-binding oligonucleotide according to any one of claims 8 to 11, wherein L is bound to the 5' end of the gapmer-type antisense oligonucleotide.

13. The lipid-binding oligonucleotide according to any one of claims 1 to 7, wherein the oligonucleotide compound is a mixed-mer type antisense oligonucleotide.

14. The lipid-binding oligonucleotide according to claim 13, wherein the mixed-mer type antisense oligonucleotide comprises at least one selected from the group consisting of 2'-modified non-crosslinked nucleotides and 2'-4'-crosslinked nucleotides.

15. The lipid-binding oligonucleotide according to claim 13 or 14, wherein the mixed-mer type antisense oligonucleotide is an oligonucleotide in which nucleosides or oligonucleotides consisting of 1 to 20 sugar-modified nucleosides and nucleosides or oligonucleotides consisting of 1 to 3 deoxyribonucleosides are alternately linked, or an oligonucleotide composed solely of sugar-modified nucleosides as the nucleosides.

16. The oligonucleotide compound according to any one of claims 13 to 15, wherein the mixed-mer type antisense oligonucleotide consists of 13 to 25 nucleosides.

17. The lipid-binding oligonucleotide compound according to any one of claims 13 to 16, wherein L is bonded to the 5' end of the mixed-mer type antisense oligonucleotide.

18. The oligonucleotide complex is a double-stranded oligonucleotide complex comprising a first oligonucleotide and a second oligonucleotide, The first oligonucleotide is a gapmer-type or mixed-type antisense oligonucleotide consisting of 7 to 100 nucleosides. The second oligonucleotide contains a sequence that enables hybridization with at least a portion of the first oligonucleotide, and consists of 4 to 100 nucleosides independently selected from deoxyribonucleosides, ribonucleosides, and sugar-modified nucleosides, and the first or second oligonucleotide is bound to L. A complex according to any one of claims 1 to 7, wherein the first oligonucleotide and the second oligonucleotide are hybridized.

19. The complex according to claim 18, wherein the second oligonucleotide comprises at least four consecutive ribonucleosides.

20. The complex according to claim 18 or 19, wherein L is bound to the 5′ end of the second oligonucleotide.

21. The oligonucleotide compound comprises a first oligonucleotide and a second oligonucleotide, The first oligonucleotide is a gapmer-type or mixed-type antisense oligonucleotide consisting of 7 to 100 nucleosides. The second oligonucleotide contains a sequence that enables hybridization with at least a portion of the first oligonucleotide, and consists of 4 to 100 nucleosides independently selected from deoxyribonucleosides, ribonucleosides, and sugar-modified nucleosides, and the first or second oligonucleotide is bound to L. The first oligonucleotide and the second oligonucleotide are linked together. A lipid-binding oligonucleotide according to any one of claims 1 to 7, wherein the first oligonucleotide and the second oligonucleotide hybridize.

22. The lipid-binding oligonucleotide according to claim 21, wherein the second oligonucleotide comprises at least four consecutive ribonucleosides.

23. The lipid-binding oligonucleotide according to claim 21 or 22, wherein L is bound to the 5' end of the second oligonucleotide.

24. The lipid-binding oligonucleotide according to any one of claims 1 to 7, wherein the oligonucleotide compound or oligonucleotide complex is selected from siRNA, aptamers, and ribozymes.

25. A pharmaceutical composition comprising a lipid-binding oligonucleotide or a complex thereof according to any one of claims 1 to 24, and a pharmacokinetically acceptable carrier.

26. A method for controlling the function of a target RNA, comprising the step of contacting a cell with a lipid-binding oligonucleotide or a complex thereof according to any one of claims 1 to 24.

27. A method for controlling the function of a target RNA in a mammal, comprising the step of administering the pharmaceutical composition described in claim 25 to the mammal.

28. A method for controlling the expression of a target gene, comprising the step of contacting a cell with a lipid-binding oligonucleotide or a complex thereof according to any one of claims 1 to 24.

29. A method for controlling the expression of a target gene in a mammal, comprising the step of administering the pharmaceutical composition described in claim 25 to the mammal.

30. A compound represented by the following formula (II). 【Chemistry 2】 (In the formula, L is a divalent group derived from a substituted or unsubstituted C1-10 alkylene group or a substituted or unsubstituted polyC1-10 alkylene glycol. R1 and R2 may each be independently a substituted or unsubstituted C5-32 alkyl group or a substituted or unsubstituted C5-32 alkenyl group, or R1 and R2 may be bonded to each other to form a ring. Each of the R3 elements may be an independent C1-10 alkyl group, or the two alkyl groups of R3 may be bonded to each other to form a ring. R4 is a 2-cyanoethyl group.

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