WO2018062413A1

WO2018062413A1 - Nucleic-acid-containing lipid nanoparticles

Info

Publication number: WO2018062413A1
Application number: PCT/JP2017/035302
Authority: WO
Inventors: 剣太朗畑中; 慎太郎細江; 隼人藪内; 香機八木
Original assignee: 協和発酵キリン株式会社
Priority date: 2016-09-28
Filing date: 2017-09-28
Publication date: 2018-04-05
Also published as: TW201813632A

Abstract

The present invention provides: nucleic-acid-containing lipid nanoparticles that include a lipid (lipid A) having a hydrophilic moiety that has one quaternary ammonium group and having three independently optionally substituted hydrocarbon groups, a lipid (lipid B) including, in the same molecule, a hydrophilic moiety that has one optionally substituted amino group or one quaternary ammonium group and a hydrophobic moiety that has two independently optionally substituted hydrocarbon groups, a lipid derivative or a fatty acid derivative of a water-soluble polymer, and a nucleic acid; and a method, etc., for producing said lipid nanoparticles.

Description

Nucleic acid-containing lipid nanoparticles

The present invention relates to, for example, nucleic acid-containing lipid nanoparticles useful as a medicament, a method for producing the lipid nanoparticles, and the like. The present invention also relates to lipids useful for producing the lipid nanoparticles.

Complexing nucleic acids with cationic lipids and other lipids as a means of efficiently delivering nucleic acids such as plasmid DNA (pDNA), antisense oligodeoxynucleic acid (ODN), and short interfering RNA (siRNA) into cells in vivo Methods for forming a body and administering the complex are known.
In Patent Document 1 and Non-Patent Document 1, as a method for producing liposomes containing nucleic acids and the like, for example, dried cationic lipid and siRNA aqueous sodium citrate solution and neutral lipid and polyethyleneglycolized phospholipid are converted into hepes buffered physiology. A solution dissolved in saline (HEPES [N- (2-Hydroxyethyl) piperazine-N '-(2-ethanesulfonic acid)] Buffered Saline, hereinafter referred to as “HBS”) and ethanol was added to diethyl ether, and water-in-oil It has been reported that a siRNA-encapsulated liposome is produced by forming a type (W / O) emulsion, mixing the solution, and treating it by a reverse phase evaporation method.
In Patent Document 2 and Non-Patent Document 2, ODN is dissolved in an aqueous citric acid solution at pH 3.8, an ethanol solution of lipid is added, and the ethanol concentration is reduced to 20 v / v% to prepare an ODN-encapsulating liposome, and a sizing membrane After the excess ethanol is removed by dialysis, the sample is further dialyzed at pH 7.5 to remove ODN adhering to the surface of the liposome to produce an ODN-encapsulating liposome.
In Patent Document 3, for example, a solution in which pDNA is dissolved in an aqueous citric acid solution and a solution in which lipids are dissolved in ethanol are mixed with a T-shaped mixer, and the ethanol concentration is lowered to 45 v / v%. Further, add citrate buffer solution to reduce ethanol concentration to 20v / v% to prepare pDNA-encapsulated liposomes, remove excess pDNA with anion exchange resin, and remove excess ethanol by ultrafiltration. A method for producing pDNA-encapsulated liposomes has been reported.
Furthermore, in Patent Document 4, pDNA-encapsulated liposomes are produced by complexing pDNA with cationic lipids as micelles in an organic solvent containing water, adding lipids, and then removing the organic solvent by dialysis. A method has been reported.
Similarly, in Patent Document 5, a pDNA-encapsulated liposome is produced by complexing pDNA as a cationic lipid and micelle in an aqueous solution of the surfactant, adding the lipid, and then removing the surfactant by dialysis. How to do it has been reported.
The cationic lipid used to form the above complex usually contains a lipophilic region containing one or more hydrocarbon groups and at least one primary amine, secondary amine, tertiary amine or quaternary ammonium group. An amphiphilic molecule having a cationic hydrophilic region, but one having two hydrocarbon groups is widely used.
On the other hand, in Patent Document 6, a cationic lipid having three hydrocarbon groups is used instead of a cationic lipid having two hydrocarbon groups, and the particle size is 82-95 nm in the same manner as in Patent Document 3. It is disclosed that lipid particles were obtained. In this document, specifically, as a cationic lipid having three hydrocarbon groups

13-B2 etc. represented by the above are disclosed.
Nucleic acid-encapsulated liposomes produced by these methods have a size of about 50-100 nm.

On the other hand, in Non-Patent Document 2, in a test using polymer micelle encapsulating dichloro (1,2-diaminocyclohexane) platinum (II) having a particle size of 30-100 nm, a polymer micelle having a smaller particle size is more likely to be in tumor. Reported higher dose and stronger anti-tumor effect. Further, in Non-Patent Document 3, it has been reported through a penetrability evaluation experiment using quantum dots that 10 nm quantum dots are more widely distributed in tumors than 100 nm quantum dots. In other words, lipid particles are required to have a smaller particle size for efficient delivery to the tumor, but any of the methods described above can efficiently prepare highly stable lipid particles having a smaller particle size. It was difficult to do.
Under these circumstances, in Patent Document 7 and Non-Patent Documents 4 to 7, a solution in which a cationic lipid, a neutral lipid, and a polyethylene glycolated lipid are dissolved in ethanol and siRNA dissolved in an acetate buffer solution are formed in a herridge bone structure. It is reported that siRNA-containing lipid nanoparticles can be obtained by mixing with microfluidics and removing ethanol. In this method, the proportion of the polar organic solvent decreases in parallel with the reverse micelle structure enclosing the nucleic acid by causing the cationic lipid in the polar organic solvent to electrostatically interact with the nucleic acid. Lipid nanoparticles in which a reverse micelle structure having a nucleic acid is coated with a lipid membrane are produced. It has been reported that the particle size of the lipid nanoparticles varies depending on the flow rate of the polar organic solvent in which the lipid is dissolved and the aqueous solution in which the nucleic acid is dissolved in the microfluidics and the PEG content. It has been reported that siRNA-encapsulated liposomes obtained by this method have exhibited drug effects in the liver even when administered subcutaneously as well as intravenously.

US Patent Application Publication No. 2013/0149374 Special table 2002-501511 gazette International Publication No. 2004/002453 International Publication No. 96/40964 US Patent Application Publication No. 2010/0041152 US Patent Application Publication No. 2012/0172411 International Publication No. 2011/140627

An object of the present invention is to provide a nucleic acid-containing lipid nanoparticle that is useful as a medicine and is more stable and smaller than conventional particles, a method for producing the lipid nanoparticle, and the like. Moreover, the objective of this invention is providing the lipid etc. which are useful for manufacture of this lipid nanoparticle.

The present invention relates to the following (1) to (87).
(1)
A lipid having a hydrophilic part having one quaternary ammonium group and three independent hydrocarbon groups which may be substituted (lipid A), one amino group which may be substituted or one quaternary ammonium group Nucleic acid, including a lipid (lipid B), a lipid derivative or fatty acid derivative of a water-soluble polymer, and a nucleic acid containing a hydrophilic moiety having a hydrophobic moiety having two independent hydrocarbon groups that may be substituted in the same molecule Contains lipid nanoparticles.
(2)
The lipid nanoparticle according to (1), wherein the number of moles of the quaternary ammonium group in lipid A is 0.01 times or more the number of moles of phosphorus atoms in the nucleic acid.
(3)
Lipid A is a lipid represented by Formula (I), Formula (II), Formula (III), Formula (IV), Formula (V ′) or Formula (V ″), or a combination thereof. The lipid nanoparticle according to (1) or (2). (Here, the formula (I), the formula (II), the formula (III), the formula (IV), the formula (V ′), and the formula (V ″) are described in the embodiments for carrying out the invention described later. The same applies hereinafter.)
(Four)
Lipid A is a lipid represented by the formula (I), and in the formula (I), one of L ¹ to L ³ is —CO—O— (CY ¹ Y ² ) _p1 — or —O—CO— ( CY ¹ Y ² ) _p1- or two or more of L ¹ to L ³ are the same or different, and -CO-O- (CY ¹ Y ² ) _p1 -or -O-CO- (CY ¹ Y ² ) _p1 - a and, R ¹ ~ R ³ is a linear or branched C15-C20 alkenyl or C9-C18 alkyl, and are the same, (3), wherein the lipid nanoparticles.
(Five)
Lipid A is a lipid represented by the formula (II), and in the formula (II), one of L ⁴ to L ⁶ is -CO-O- (CY ⁸ Y ⁹ ) _p4- , -O-CO- ( CY ⁸ Y ⁹ ) _p4 -or -O-CO- (CY ¹⁰ Y ¹¹ ) _p5 -O- (CY ¹² Y ¹³ ) _p6- or two or more of L ⁴ to L ⁶ are the same or different -CO-O- (CY ⁸ Y ⁹ ) _p4- , -O-CO- (CY ⁸ Y ⁹ ) _p4- , or -O-CO- (CY ¹⁰ Y ¹¹ ) _p5 -O- (CY ¹² Y ¹³ The lipid nanoparticles according to (3), which are _p6- and R ⁴ to R ⁶ are linear or branched C15-C20 alkenyl or C9-C18 alkyl and are the same.
(6)
Lipid A is a lipid represented by the formula (III), and in the formula (III), one of L ⁸ to L ¹⁰ is -CO-O- (CY ³⁹ Y ⁴⁰ ) _p18 -or -O-CO- ( CY ³⁹ Y ⁴⁰ ) _p18- , or two or more of L ⁸ to L ¹⁰ are the same or different -CO-O- (CY ³⁹ Y ⁴⁰ ) _p18 or -O-CO- (CY ³⁹ Y ⁴⁰ ) The lipid nanoparticle according to (3), which is _p18- and R ⁷ to R ⁹ are linear or branched C15-C20 alkenyl or C9-C18 alkyl.
(7)
Lipid A is a lipid represented by the formula (IV), and in the formula (IV), L ¹³ is -CO-O- (CY ⁸⁴ Y ⁸⁵ ) _p38 -or -O-CO- (CY ⁸⁴ Y ⁸⁵ ) _p38 -Or one of L ¹⁴ and L ¹⁵ is -CO-O- (CY ⁹¹ Y ⁹² ) _p41 -or -O-CO- (CY ⁹¹ Y ⁹² ) _p41- , or L ¹³ is -CO -O- (CY ⁸⁴ Y ⁸⁵ ) _{p38 -or} -O-CO- (CY ⁸⁴ Y ⁸⁵ ) _p38- and one of L ¹⁴ and L ¹⁵ is -CO-O- (CY ⁹¹ Y ⁹² ) _p41 -Or-O-CO- (CY ⁹¹ Y ⁹² ) _p41- or L ¹⁴ and L ¹⁵ are the same or different -CO-O- (CY ⁹¹ Y ⁹² ) _{p41 -or} -O-CO- (CY ⁹¹ Y ⁹² ) _p41- or L ¹³ is -CO-O- (CY ⁸⁴ Y ⁸⁵ ) _p38 -or -O-CO- (CY ⁸⁴ Y ⁸⁵ ) _p38- and L ¹⁴ and L ¹⁵ Are the same or different and are -CO-O- (CY ⁹¹ Y ⁹² ) _p41 -or -O-CO- (CY ⁹¹ Y ⁹² ) _p41- , and R ¹⁰ to R ¹² are linear or branched C15 The lipid nanoparticle according to (3), which is -C20 alkenyl or C9-C18 alkyl.
(8)
Lipid A is a lipid represented by the formula (V ′), and in the formula (V ′), one of L ¹⁷ to L ¹⁹ is —CO—O— or —O—, or L ¹⁷ to L ¹⁹ Two or more of them are the same or different and are —CO—O— or —O—, and R ¹³ to R ¹⁵ are linear or branched C15-C20 alkenyl or C9-C18 alkyl, (3) The lipid nanoparticles described.
(9)
10. The lipid nanoparticle according to any one of (1) to (8), wherein the content of the water-soluble polymer lipid derivative or fatty acid derivative is 0.005 times the molar amount of the total lipid or more.
(Ten)
(1) to (9), wherein the water-soluble polymer lipid derivative or the fatty acid derivative of the water-soluble polymer is selected from the group consisting of polyethylene glycol, polyglycerin, polyethyleneimine, polyvinyl alcohol, polyacrylic acid and polyacrylamide. The lipid nanoparticle according to any one of the above.
(11)
The lipid nanoparticles according to any one of (1) to (10), wherein the ratio of the number of moles of total lipid to the number of moles of nucleic acid (number of moles of total lipid / number of moles of nucleic acid) is 50 or more.
(12)
The lipid nanoparticle according to any one of (1) to (11), wherein the nucleic acid is a nucleic acid having an action of suppressing the expression of a target gene using RNA interference (RNAi).
(13)
The lipid nanoparticle according to (12), wherein the target gene is a gene associated with tumor or inflammation.
(14)
The lipid nanoparticle according to any one of (1) to (13), wherein the content of lipid B is 0.1 times or more by mole of the total number of moles of lipid.
(15)
The number of moles of the amino group or quaternary ammonium group in the hydrophilic part in the lipid B is 0.01 times or more the mole number of the phosphorus atom in the nucleic acid, according to any one of (1) to (14) Lipid nanoparticles.
(16)
Lipid B is represented by formula (CL-I), formula (CL-II), formula (CL-III), formula (CL-IV), formula (CL-V), formula (CL-VI), formula (CL- (VII), formula (CL-VIII), formula (CL-IX), formula (CL-X), formula (CL-XI), formula (CL-XII), formula (CL-XIII), formula (CL-XIV ), A formula (CL-XV), a formula (CL-XVI), or a lipid represented by the formula (CL-XVII) or a combination thereof, according to any one of (1) to (15) Lipid nanoparticles. (Where, formula (CL-I), formula (CL-II), formula (CL-III), formula (CL-IV), formula (CL-V), formula (CL-VI), formula (CL- (VII), formula (CL-VIII), formula (CL-IX), formula (CL-X), formula (CL-XI), formula (CL-XII), formula (CL-XIII), formula (CL-XIV ), Formula (CL-XV), formula (CL-XVI), and formula (CL-XVII) are represented by the structures described in the modes for carrying out the invention to be described later. )
(17)
The lipid nanoparticle according to any one of (1) to (16), further comprising a neutral lipid.
(18)
The lipid nanoparticle according to (17), wherein the content of neutral lipid is 0.05 times or more by mole of the total number of moles of lipid.
(19)
The lipid nanoparticle according to (17) or (18), wherein the neutral lipid is selected from the group consisting of phospholipid, sterol, glyceroglycolipid, sphingoglycolipid and sphingoid.
(20)
The lipid nanoparticles according to any one of (1) to (19), wherein the average particle size is 20 to 65 nm.
(twenty one)
A method for producing nucleic acid-containing lipid nanoparticles comprising:
(a) a lipid having a hydrophilic part having one quaternary ammonium group and three independent hydrocarbon groups optionally substituted in a mixed solvent of water and an organic solvent miscible with water (lipid A) A lipid having a hydrophilic part having one amino group which may be substituted or one quaternary ammonium group and a hydrophobic part having two independent hydrocarbon groups which may be substituted (lipid B) and a nucleic acid to prepare a first lipid solution containing lipid A and nucleic acid,
(b) adding a second lipid solution containing a lipid derivative or fatty acid derivative of a water-soluble polymer to the first lipid solution to prepare a third lipid solution; and
(c) The production method comprising a step of adding water or an aqueous buffer solution to the third lipid solution.
(twenty two)
The production method according to (21), comprising mixing a lipid derivative or fatty acid derivative of a water-soluble polymer, and a neutral lipid together with lipid A, lipid B, and nucleic acid in step (a).
(twenty three)
The second lipid solution in step (b) is further neutral lipid and / or a hydrophilic part having one amino group or one quaternary ammonium group which may be substituted and an independent which may be substituted The production method according to (21), comprising a lipid (lipid B) containing a hydrophobic moiety having two hydrocarbon groups in the same molecule.
(twenty four)
The production method according to any one of (21) to (23), wherein the organic solvent miscible with water is alcohol, dimethyl sulfoxide, tetrahydrofuran, acetone, acetonitrile, or a mixture thereof.
(twenty five)
The production method according to any one of (21) to (23), wherein the organic solvent miscible with water is an alcohol.
(26)
The production method according to (24) or (25), wherein the alcohol is methanol, ethanol, propanol, butanol or a mixture thereof.
(27)
The content of the organic solvent miscible with water in the third lipid solution obtained in step (b) is 50% (v / v) or more with respect to the solution, (21) to (26) The production method according to any one of the above.
(28)
In step (c), the concentration of the organic solvent miscible with water in the third lipid solution obtained in step (b) is reduced to 50% (v / v) or less by adding water or an aqueous buffer solution. (21) The production method according to any one of (27) to (27).
(29)
The production method according to (28), wherein the concentration of the organic solvent miscible with water in the third lipid solution obtained in the step (b) is 50% (v / v) or less within 1 minute.
(30)
(21) The production method according to any one of (21) to (29), wherein the number of moles of the quaternary ammonium group in lipid A is 0.01 times or more the number of moles of phosphorus atoms in the nucleic acid.
(31)
The content of the organic solvent miscible with water in the first lipid solution obtained in the step (a) is 50% (v / v) or more with respect to the solution, (21) to (30) The production method according to any one of the above.
(32)
The production method according to any one of (21) to (31), wherein the aqueous buffer solution is a phosphate buffer aqueous solution, a citrate buffer aqueous solution, or an acetate buffer aqueous solution.
(33)
The ratio of the number of moles of total lipid to the number of moles of nucleic acid in the third lipid solution (number of moles of total lipid / number of moles of nucleic acid) is 50 or more, according to any one of (21) to (32) The manufacturing method as described.
(34)
The production method according to any one of (21) to (33), wherein the nucleic acid is a nucleic acid having an action of suppressing the expression of a target gene using RNA interference (RNAi).
(35)
The production method according to (34), wherein the target gene is a gene associated with tumor or inflammation.
(36)
Lipid A is a lipid represented by Formula (I), Formula (II), Formula (III), Formula (IV), Formula (V ′) or Formula (V ″), or a combination thereof. (21) The production method according to any one of (35).
(37)
Lipid A is a lipid represented by the formula (I), and in the formula (I), one of L ¹ to L ³ is —CO—O— (CY ¹ Y ² ) _p1 — or —O—CO— ( CY ¹ Y ² ) _p1- or two or more of L ¹ to L ³ are the same or different, and -CO-O- (CY ¹ Y ² ) _p1 -or -O-CO- (CY ¹ Y ² ) The production method according to (36), wherein _p ^1- and R ¹ to R ³ are linear or branched C15-C20 alkenyl or C9-C18 alkyl, and are the same.
(38)
Lipid A is a lipid represented by the formula (II), and in the formula (II), one of L ⁴ to L ⁶ is -CO-O- (CY ⁸ Y ⁹ ) _p4- , -O-CO- ( CY ⁸ Y ⁹ ) _p4 -or -O-CO- (CY ¹⁰ Y ¹¹ ) _p5 -O- (CY ¹² Y ¹³ ) _p6- or two or more of L ⁴ to L ⁶ are the same or different -CO-O- (CY ⁸ Y ⁹ ) _p4- , -O-CO- (CY ⁸ Y ⁹ ) _p4- , or -O-CO- (CY ¹⁰ Y ¹¹ ) _p5 -O- (CY ¹² Y ¹³ ) The production method according to (36), wherein _{p 6-} and R ⁴ to R ⁶ are linear or branched C15-C20 alkenyl or C9-C18 alkyl, and are the same.
(39)
Lipid A is a lipid represented by the formula (III), and in the formula (III), one of L ⁸ to L ¹⁰ is -CO-O- (CY ³⁹ Y ⁴⁰ ) _p18 -or -O-CO- ( CY ³⁹ Y ⁴⁰ ) _p18- , or two or more of L ⁸ to L ¹⁰ are the same or different -CO-O- (CY ³⁹ Y ⁴⁰ ) _p18 or -O-CO- (CY ³⁹ Y ⁴⁰ ) _{pi 8} - a and, R ⁷ ~ R ⁹ is a linear or branched C15-C20 alkenyl or C9-C18 alkyl, (36) the method according.
(40)
Lipid A is a lipid represented by the formula (IV), and in the formula (IV), L ¹³ is -CO-O- (CY ⁸⁴ Y ⁸⁵ ) _p38 -or -O-CO- (CY ⁸⁴ Y ⁸⁵ ) _p38 -Or one of L ¹⁴ and L ¹⁵ is -CO-O- (CY ⁹¹ Y ⁹² ) _p41 -or -O-CO- (CY ⁹¹ Y ⁹² ) _p41- , or L ¹³ is -CO -O- (CY ⁸⁴ Y ⁸⁵ ) _{p38 -or} -O-CO- (CY ⁸⁴ Y ⁸⁵ ) _p38- and one of L ¹⁴ and L ¹⁵ is -CO-O- (CY ⁹¹ Y ⁹² ) _p41 -Or-O-CO- (CY ⁹¹ Y ⁹² ) _p41- or L ¹⁴ and L ¹⁵ are the same or different -CO-O- (CY ⁹¹ Y ⁹² ) _{p41 -or} -O-CO- (CY ⁹¹ Y ⁹² ) _p41- or L ¹³ is -CO-O- (CY ⁸⁴ Y ⁸⁵ ) _p38 -or -O-CO- (CY ⁸⁴ Y ⁸⁵ ) _p38- and L ¹⁴ and L ¹⁵ Are the same or different and are -CO-O- (CY ⁹¹ Y ⁹² ) _p41 -or -O-CO- (CY ⁹¹ Y ⁹² ) _p41- , and R ¹⁰ to R ¹² are linear or branched C15 The production method of (36), which is —C20 alkenyl or C9-C18 alkyl.
(41)
Lipid A is a lipid represented by the formula (V ′), and in the formula (V ′), one of L ¹⁷ to L ¹⁹ is —CO—O— or —O—, or L ¹⁷ to L ¹⁹ Two or more of them are the same or different and are —CO—O— or —O—, and R ¹³ to R ¹⁵ are linear or branched C15-C20 alkenyl or C9-C18 alkyl, (36) The manufacturing method as described.
(42)
Lipid B is represented by formula (CL-I), formula (CL-II), formula (CL-III), formula (CL-IV), formula (CL-V), formula (CL-VI), formula (CL- (VII), formula (CL-VIII), formula (CL-IX), formula (CL-X), formula (CL-XI), formula (CL-XII), formula (CL-XIII), formula (CL-XIV ), A formula (CL-XV), a formula (CL-XVI), or a lipid represented by the formula (CL-XVII) or a combination thereof, the production method according to any one of (21) to (41) .
(43)
A method for producing nucleic acid-containing lipid nanoparticles comprising:
(a) a lipid having a hydrophilic part having one quaternary ammonium group and three independent hydrocarbon groups optionally substituted in a mixed solvent of water and an organic solvent miscible with water (lipid A) A lipid having a hydrophilic part having one amino group which may be substituted or one quaternary ammonium group and a hydrophobic part having two independent hydrocarbon groups which may be substituted (lipid A step of mixing B), a nucleic acid, and a lipid derivative or fatty acid derivative of a water-soluble polymer to prepare a first lipid solution containing the lipid and the nucleic acid; and
(b) The production method comprising a step of adding water or an aqueous buffer solution to the first lipid solution.
(44)
In step (a), it has a neutral lipid, and / or a hydrophilic part having one amino group or one quaternary ammonium group which may be substituted, and two independent hydrocarbon groups which may be substituted The production method according to (43), comprising mixing a lipid (lipid B) containing a hydrophobic part in the same molecule.
(45)
The production method according to (43) or (44), wherein the water-miscible organic solvent is alcohol, dimethyl sulfoxide, tetrahydrofuran, acetone, acetonitrile or a mixture thereof.
(46)
The production method according to (43) or (44), wherein the water-miscible organic solvent is an alcohol.
(47)
The production method according to (45) or (46), wherein the alcohol is methanol, ethanol, propanol, butanol or a mixture thereof.
(48)
The content of the water-miscible organic solvent in the first lipid solution obtained in the step (a) is 50% (v / v) or more with respect to the solution (43) to (47) The production method according to any one of the above.
(49)
In step (b), by adding water or an aqueous buffer solution, the concentration of the organic solvent miscible with water in the first lipid solution obtained in step (a) is 50% (v / v) or less. The production method according to any one of (43) to (48).
(50)
The production method according to (49), wherein the concentration of the organic solvent miscible with water in the first lipid solution obtained in the step (a) is 50% (v / v) or less within 1 minute.
(51)
The production according to any one of (43) to (50), wherein the number of moles of the quaternary ammonium group in lipid A is 0.01 times or more the number of moles of phosphate atoms in the nucleic acid Method.
(52)
The production method according to any one of (43) to (51), wherein the aqueous buffer solution is a phosphate buffer aqueous solution, a citrate buffer aqueous solution, or an acetate buffer aqueous solution.
(53)
The ratio of the number of moles of total lipid to the number of moles of nucleic acid used in step (a) (the number of moles of total lipid / the number of moles of nucleic acid) is 50 or more, according to any one of (43) to (52) Manufacturing method.
(54)
The production method according to any one of (43) to (53), wherein the nucleic acid is a nucleic acid having an action of suppressing the expression of a target gene using RNA interference (RNAi).
(55)
(54) The production method according to (54), wherein the target gene is a gene associated with tumor or inflammation.
(56)
Lipid A is a lipid represented by Formula (I), Formula (II), Formula (III), Formula (IV), Formula (V ′) or Formula (V ″), or a combination thereof. (43) The production method according to any one of (55).
(57)
Lipid A is a lipid represented by the formula (I), and in the formula (I), one of L ¹ to L ³ is —CO—O— (CY ¹ Y ² ) _p1 — or —O—CO— ( CY ¹ Y ² ) _p1- or two or more of L ¹ to L ³ are the same or different, and -CO-O- (CY ¹ Y ² ) _p1 -or -O-CO- (CY ¹ Y ² ) The production method according to (56), wherein _p ^1- and R ¹ to R ³ are linear or branched C15-C20 alkenyl or C9-C18 alkyl, and are the same.
(58)
Lipid A is a lipid represented by the formula (II), and in the formula (II), one of L ⁴ to L ⁶ is -CO-O- (CY ⁸ Y ⁹ ) _p4- , -O-CO- ( CY ⁸ Y ⁹ ) _p4 -or -O-CO- (CY ¹⁰ Y ¹¹ ) _p5 -O- (CY ¹² Y ¹³ ) _p6- or two or more of L ⁴ to L ⁶ are the same or different -CO-O- (CY ⁸ Y ⁹ ) _p4- , -O-CO- (CY ⁸ Y ⁹ ) _p4- , or -O-CO- (CY ¹⁰ Y ¹¹ ) _p5 -O- (CY ¹² Y ¹³ ) _p6 - a and, R ⁴ ~ R ⁶ is a linear or branched C15-C20 alkenyl or C9-C18 alkyl, and are the same, (56) the method according.
(59)
Lipid A is a lipid represented by the formula (III), and in the formula (III), one of L ⁸ to L ¹⁰ is -CO-O- (CY ³⁹ Y ⁴⁰ ) _p18 -or -O-CO- ( CY ³⁹ Y ⁴⁰ ) _p18- , or two or more of L ⁸ to L ¹⁰ are the same or different -CO-O- (CY ³⁹ Y ⁴⁰ ) _p18 or -O-CO- (CY ³⁹ Y ⁴⁰ ) The production method according to (56), wherein _p18- and R ⁷ to R ⁹ are linear or branched C15-C20 alkenyl or C9-C18 alkyl.
(60)
Lipid A is a lipid represented by the formula (IV), and in the formula (IV), L ¹³ is -CO-O- (CY ⁸⁴ Y ⁸⁵ ) _p38 -or -O-CO- (CY ⁸⁴ Y ⁸⁵ ) _p38 -Or one of L ¹⁴ and L ¹⁵ is -CO-O- (CY ⁹¹ Y ⁹² ) _p41 -or -O-CO- (CY ⁹¹ Y ⁹² ) _p41- , or L ¹³ is -CO -O- (CY ⁸⁴ Y ⁸⁵ ) _{p38 -or} -O-CO- (CY ⁸⁴ Y ⁸⁵ ) _p38- and one of L ¹⁴ and L ¹⁵ is -CO-O- (CY ⁹¹ Y ⁹² ) _p41 -Or-O-CO- (CY ⁹¹ Y ⁹² ) _p41- or L ¹⁴ and L ¹⁵ are the same or different -CO-O- (CY ⁹¹ Y ⁹² ) _{p41 -or} -O-CO- (CY ⁹¹ Y ⁹² ) _p41- or L ¹³ is -CO-O- (CY ⁸⁴ Y ⁸⁵ ) _p38 -or -O-CO- (CY ⁸⁴ Y ⁸⁵ ) _p38- and L ¹⁴ and L ¹⁵ Are the same or different and are -CO-O- (CY ⁹¹ Y ⁹² ) _p41 -or -O-CO- (CY ⁹¹ Y ⁹² ) _p41- , and R ¹⁰ to R ¹² are linear or branched C15 The production method according to (56), which is -C20 alkenyl or C9-C18 alkyl.
(61)
Lipid A is a lipid represented by the formula (V ′), and in the formula (V ′), one of L ¹⁷ to L ¹⁹ is —CO—O— or —O—, or L ¹⁷ to L ¹⁹ Two or more of them are the same or different and are —CO—O— or —O—, and R ¹³ to R ¹⁵ are linear or branched C15-C20 alkenyl or C9-C18 alkyl, (56) The manufacturing method as described.
(62)
Lipid B is represented by formula (CL-I), formula (CL-II), formula (CL-III), formula (CL-IV), formula (CL-V), formula (CL-VI), formula (CL- (VII), formula (CL-VIII), formula (CL-IX), formula (CL-X), formula (CL-XI), formula (CL-XII), formula (CL-XIII), formula (CL-XIV ), A lipid represented by formula (CL-XV), a formula (CL-XVI), or a lipid represented by formula (CL-XVII) or a combination thereof, (43) to (61) The manufacturing method in any one of.
(63)
(21) A nucleic acid-containing lipid nanoparticle obtained by the production method according to any one of (62).
(64)
Nucleic acid containing, using a lipid membrane (lipid A) having a hydrophilic part having one quaternary ammonium group and three independent hydrocarbon groups which may be substituted in the lipid membrane constituting the nucleic acid containing lipid nanoparticle A method of stabilizing lipid nanoparticles.
(65)
The method according to (64), wherein the number of moles of the quaternary ammonium group in lipid A is 0.01 times or more the mole number of phosphorus atoms in the nucleic acid.
(66)
Lipid A is a lipid represented by Formula (I), Formula (II), Formula (III), Formula (IV), Formula (V ′) or Formula (V ″), or a combination thereof. (64) or the method according to (65).
(67)
Lipid A is a lipid represented by the formula (I), and in the formula (I), one of L ¹ to L ³ is —CO—O— (CY ¹ Y ² ) _p1 — or —O—CO— ( CY ¹ Y ² ) _p1- or two or more of L ¹ to L ³ are the same or different, and -CO-O- (CY ¹ Y ² ) _p1 -or -O-CO- (CY ¹ Y ² ) The method according to (66), wherein _p ^1- and R ¹ to R ³ are linear or branched C15-C20 alkenyl or C9-C18 alkyl, and are the same.
(68)
Lipid A is a lipid represented by the formula (II), and in the formula (II), one of L ⁴ to L ⁶ is -CO-O- (CY ⁸ Y ⁹ ) _p4- , -O-CO- ( CY ⁸ Y ⁹ ) _p4 -or -O-CO- (CY ¹⁰ Y ¹¹ ) _p5 -O- (CY ¹² Y ¹³ ) _p6- or two or more of L ⁴ to L ⁶ are the same or different -CO-O- (CY ⁸ Y ⁹ ) _p4- , -O-CO- (CY ⁸ Y ⁹ ) _p4- , or -O-CO- (CY ¹⁰ Y ¹¹ ) _p5 -O- (CY ¹² Y ¹³ ) The method according to (66), wherein _p6- and R ⁴ to R ⁶ are linear or branched C15-C20 alkenyl or C9-C18 alkyl, and are the same.
(69)
Lipid A is a lipid represented by the formula (III), and in the formula (III), one of L ⁸ to L ¹⁰ is -CO-O- (CY ³⁹ Y ⁴⁰ ) _p18 -or -O-CO- ( CY ³⁹ Y ⁴⁰ ) _p18- , or two or more of L ⁸ to L ¹⁰ are the same or different -CO-O- (CY ³⁹ Y ⁴⁰ ) _p18 or -O-CO- (CY ³⁹ Y ⁴⁰ ) The method according to (66), wherein _p18— and R ⁷ to R ⁹ are linear or branched C15-C20 alkenyl or C9-C18 alkyl.
(70)
Lipid A is a lipid represented by the formula (IV), and in the formula (IV), L ¹³ is -CO-O- (CY ⁸⁴ Y ⁸⁵ ) _p38 -or -O-CO- (CY ⁸⁴ Y ⁸⁵ ) _p38 -Or one of L ¹⁴ and L ¹⁵ is -CO-O- (CY ⁹¹ Y ⁹² ) _p41 -or -O-CO- (CY ⁹¹ Y ⁹² ) _p41- , or L ¹³ is -CO -O- (CY ⁸⁴ Y ⁸⁵ ) _{p38 -or} -O-CO- (CY ⁸⁴ Y ⁸⁵ ) _p38- and one of L ¹⁴ and L ¹⁵ is -CO-O- (CY ⁹¹ Y ⁹² ) _p41 -Or-O-CO- (CY ⁹¹ Y ⁹² ) _p41- or L ¹⁴ and L ¹⁵ are the same or different -CO-O- (CY ⁹¹ Y ⁹² ) _{p41 -or} -O-CO- (CY ⁹¹ Y ⁹² ) _p41- or L ¹³ is -CO-O- (CY ⁸⁴ Y ⁸⁵ ) _p38 -or -O-CO- (CY ⁸⁴ Y ⁸⁵ ) _p38- and L ¹⁴ and L ¹⁵ Are the same or different and are -CO-O- (CY ⁹¹ Y ⁹² ) _p41 -or -O-CO- (CY ⁹¹ Y ⁹² ) _p41- , and R ¹⁰ to R ¹² are linear or branched C15 The method according to (66), which is —C20 alkenyl or C9-C18 alkyl.
(71)
Lipid A is a lipid represented by the formula (V ′), and in the formula (V ′), one of L ¹⁷ to L ¹⁹ is —CO—O— or —O—, or L ¹⁷ to L ¹⁹ Two or more of them are the same or different and are —CO—O— or —O—, and R ¹³ to R ¹⁵ are linear or branched C15-C20 alkenyl or C9-C18 alkyl, (66) The method described.
(72)
One quaternary ammonium selected from the group consisting of lipids of formula (I), formula (II), formula (III), formula (IV), formula (V ′) and formula (V ″) A lipid having a hydrophilic part having a group and three independent hydrocarbon groups which may be substituted (lipid A).
(73)
Lipid A is a lipid represented by the formula (I), and in the formula (I), one of L ¹ to L ³ is —CO—O— (CY ¹ Y ² ) _p1 — or —O—CO— ( CY ¹ Y ² ) _p1- or two or more of L ¹ to L ³ are the same or different, and -CO-O- (CY ¹ Y ² ) _p1 -or -O-CO- (CY ¹ Y ² ) _p1 - a is a R ¹ ~ R ³ is a linear or branched C15-C20 alkenyl or C9-C18 alkyl, and are the same, (72) a lipid according.
(74)
Lipid A is a lipid represented by the formula (II), and in the formula (II), one of L ⁴ to L ⁶ is -CO-O- (CY ⁸ Y ⁹ ) _p4- , -O-CO- ( CY ⁸ Y ⁹ ) _p4 -or -O-CO- (CY ¹⁰ Y ¹¹ ) _p5 -O- (CY ¹² Y ¹³ ) _p6- or two or more of L ⁴ to L ⁶ are the same or different -CO-O- (CY ⁸ Y ⁹ ) _p4- , -O-CO- (CY ⁸ Y ⁹ ) _p4- , or -O-CO- (CY ¹⁰ Y ¹¹ ) _p5 -O- (CY ¹² Y ¹³ The lipid according to (72), which is _p6- , and R ⁴ to R ⁶ are linear or branched C15-C20 alkenyl or C9-C18 alkyl, and are the same.
(75)
Lipid A is a lipid represented by the formula (III), and in the formula (III), one of L ⁸ to L ¹⁰ is -CO-O- (CY ³⁹ Y ⁴⁰ ) _p18 -or -O-CO- ( CY ³⁹ Y ⁴⁰ ) _p18- , or two or more of L ⁸ to L ¹⁰ are the same or different -CO-O- (CY ³⁹ Y ⁴⁰ ) _p18 or -O-CO- (CY ³⁹ Y ⁴⁰ ) The lipid according to (72), which is _p18- and R ⁷ to R ⁹ are linear or branched C15-C20 alkenyl or C9-C18 alkyl.
(76)
Lipid A is a lipid represented by the formula (IV), and in the formula (IV), L ¹³ is -CO-O- (CY ⁸⁴ Y ⁸⁵ ) _p38 -or -O-CO- (CY ⁸⁴ Y ⁸⁵ ) _p38 -Or one of L ¹⁴ and L ¹⁵ is -CO-O- (CY ⁹¹ Y ⁹² ) _p41 -or -O-CO- (CY ⁹¹ Y ⁹² ) _p41- , or L ¹³ is -CO -O- (CY ⁸⁴ Y ⁸⁵ ) _{p38 -or} -O-CO- (CY ⁸⁴ Y ⁸⁵ ) _p38- and one of L ¹⁴ and L ¹⁵ is -CO-O- (CY ⁹¹ Y ⁹² ) _p41 -Or-O-CO- (CY ⁹¹ Y ⁹² ) _p41- or L ¹⁴ and L ¹⁵ are the same or different -CO-O- (CY ⁹¹ Y ⁹² ) _{p41 -or} -O-CO- (CY ⁹¹ Y ⁹² ) _p41- or L ¹³ is -CO-O- (CY ⁸⁴ Y ⁸⁵ ) _p38 -or -O-CO- (CY ⁸⁴ Y ⁸⁵ ) _p38- and L ¹⁴ and L ¹⁵ Are the same or different and are -CO-O- (CY ⁹¹ Y ⁹² ) _p41 -or -O-CO- (CY ⁹¹ Y ⁹² ) _p41- , and R ¹⁰ to R ¹² are linear or branched C15 The lipid according to (72), which is -C20 alkenyl or C9-C18 alkyl.
(77)
Lipid A is a lipid represented by the formula (V ′), and in the formula (V ′), one of L ¹⁷ to L ¹⁹ is —CO—O— or —O—, or L ¹⁷ to L ¹⁹ Two or more of them are the same or different and are —CO—O— or —O—, and R ¹³ to R ¹⁵ are linear or branched C15-C20 alkenyl or C9-C18 alkyl, (72) The lipids described.
(78)
(1) A method for introducing a nucleic acid into a cell using the nucleic acid-containing lipid nanoparticle according to any one of (20) and (63).
(79)
The method according to (78), wherein the cell is a cell in a mammalian tumor or inflammation site.
(80)
The method according to (78) or (79), wherein the cell is a cell in the liver, stomach, lung, kidney, pancreas or spleen of a mammal.
(81)
The method according to any one of (78) to (80), wherein the method of introduction into cells is a method of introduction into cells by intravenous administration or subcutaneous administration.
(82)
(1) A method for treating cancer or inflammatory disease, comprising administering the nucleic acid-containing lipid nanoparticles according to any one of (20) and (63) to a mammal.
(83)
The method according to (82), wherein the method of administration is intravenous administration or subcutaneous administration.
(84)
(1) A pharmaceutical comprising the nucleic acid-containing lipid nanoparticle according to any one of (20) and (63).
(85)
The medicament according to (84), which is for intravenous administration or subcutaneous administration.
(86)
A therapeutic agent for cancer or inflammatory disease comprising the nucleic acid-containing lipid nanoparticles according to any one of (1) to (20) and (63).
(87)
The therapeutic agent according to (86), which is for intravenous administration or subcutaneous administration.

According to the present invention, it is possible to provide a nucleic acid-containing lipid nanoparticle that is useful as a medicine and is more stable and smaller than conventional particles, a method for producing the lipid nanoparticle, and the like. Further, according to the present invention, lipids and the like useful for the production of the lipid nanoparticles can be provided.

Hereinafter, embodiments for carrying out the present invention will be described in detail. Note that the embodiments described below do not limit the present invention.
The nucleic acid-containing lipid nanoparticle of the present invention comprises a lipid having a hydrophilic part having one quaternary ammonium group and three independently substituted hydrocarbon groups (lipid A), a lipid derivative of a water-soluble polymer, or Includes fatty acid derivatives, as well as nucleic acids. In the present invention, a lipid derivative or fatty acid derivative of a water-soluble polymer using a hydrophilic part having one quaternary ammonium group and a lipid having three independent hydrocarbon groups which may be substituted (lipid A) In addition to nucleic acid-containing lipid nanoparticles together with nucleic acids, nucleic acid-containing lipid nanoparticles that are more excellent in physicochemical stability and physiological activity can be obtained.

In the present invention, a lipid having a hydrophilic part having one quaternary ammonium group and three independent hydrocarbon groups which may be substituted (lipid A) includes one quaternary ammonium group as a hydrophilic part in the molecule. The molecule is not particularly limited as long as it is a molecule having three independent hydrocarbon groups which may be substituted and may be substituted, for example, represented by the following structural formulas (A) to (C). In the following structural formulas (A) to (C), “Hydrophilic unit” represents a hydrophilic unit having one quaternary ammonium group, and three “Hydrophobic units” are substituted. May represent three independent hydrocarbon groups.

The quaternary ammonium group that constitutes the “hydrophilic part (Hydrophilic Unit)” has 0 to 3 of the four bonds, and any of the hydrocarbon groups that form the “hydrophobic part (Hydrophobic Unit)” 0 to The three bonds are bonded, and the remaining bond is bonded to an optionally substituted chain and / or cyclic hydrocarbon group. The optionally substituted chain and / or cyclic hydrocarbon group constituting the “hydrophilic unit” may be any group consisting of a carbon atom and a hydrogen atom. Those having 10 carbon atoms are preferable, those having 1 to 6 carbon atoms are more preferable, and those having 1 to 3 carbon atoms are more preferable.

In addition, the “hydrophilic unit” refers to one or more ethers, esters, amides and the like via carbon atoms in the chain and / or cyclic hydrocarbon group which may be substituted. You may have. Furthermore, examples of the substituent in the linear and / or cyclic hydrocarbon group which may be substituted include carbamate, amino, monoalkylamino, dialkylamino, pyrrolidin-2-yl, pyrrolidin-3-yl, and piperidine-2. -Yl, piperidin-3-yl, piperidin-4-yl, morpholin-2-yl, morpholin-3-yl, hydroxy, alkoxy, alkoxycarbonyl, hydroxycarbonyl, carbamoyl, monoalkylcarbamoyl, dialkylcarbamoyl, nitro, cyano, Examples include fluoro, chloro, or bromo.

In addition, the hydrocarbon group forming the “hydrophobic unit” may be any group consisting of 8 to 24 carbon atoms and hydrogen atoms. The hydrocarbon group can be classified from the viewpoint of topology, and examples thereof include a linear hydrocarbon group, a branched hydrocarbon group, or a cyclic hydrocarbon group (for example, a cholesteryl group). A branched or branched hydrocarbon group is preferred. In addition, the hydrocarbon group can be classified by the presence or absence of an unsaturated bond (double bond or triple bond), and the hydrocarbon group having an unsaturated bond can also be classified by the presence or absence of aromaticity. As the hydrocarbon group, a hydrocarbon group consisting of only a saturated bond (alkyl) or a hydrocarbon group having an unsaturated bond and having no aromaticity (for example, alkenyl or alkynyl) is preferable. The hydrocarbon group in lipid A is preferably linear or branched C8-C24 alkyl, C8-C24 alkenyl, C8-C24 alkynyl.

The hydrocarbon groups forming the “Hydrophobic Unit” may be directly bonded to the quaternary ammonium group of the “Hydrophilic Unit”, or the bonds such as ether, ester, amide, etc. It may be bonded to a quaternary ammonium group via a chain and / or cyclic hydrocarbon group which may have a substituent constituting “part”. In addition, as shown in structural formulas (B) and (C), two or three hydrocarbon groups forming “Hydrophobic Unit” are bonded via a carbon atom, and the carbon atom Is a chain and / or cyclic carbonization which may have a quaternary ammonium group of “Hydrophilic Unit” directly, or a bond constituting ether, ester, amide, etc. and a substituent constituting “hydrophilic portion” It may be bonded to a quaternary ammonium group through a hydrogen group or the like.

As lipid A,
Formula (I)

(Where
R ¹ to R ³ are the same or different and may be linear or branched C8-C24 alkyl, C8-C24 alkenyl or C8-C24 alkynyl,
L ¹ to L ³ are the same or different and do not exist, -Z ^1- (CY ¹ Y ² ) _p1 -or -Z ^2- (CY ³ Y ⁴ ) _p2 -Z ^3- (CY ⁵ Y ⁶ ) _p3- (Wherein Y ¹ to Y ⁶ are the same or different hydrogen atoms or optionally substituted C1-C4 alkyl, Z ¹ to Z ³ are the same or different, and —O—, —NY ^7A —, -CO-O -, - O- CO -, - CO-NY 7B -, - NY 7C -CO- or -NY ^7D -CO-0- and is (wherein, Y ^7A ~ Y ^7D are the same or different and A hydrogen atom or an optionally substituted C1-C4 alkyl), p ¹ to p ³ are the same or different and are an integer of 1 to 5),
X ¹ is an optionally substituted C1-C4 alkyl,
A ¹ is a pharmaceutically acceptable anion)
Formula (II)

(Where
R ⁴ to R ⁶ are the same or different and may be linear or branched C8-C24 alkyl, C8-C24 alkenyl or C8-C24 alkynyl,
L ⁴ ~ L ⁶ is absent the same or ^{^{different, -Z 4 - (CY 8 Y}} 9) p4 - or ^{^{^{-Z 5 - (CY 10 Y 11}}} ) p5 -Z 6 - (CY 12 Y 13) p6 - (Wherein Y ⁸ to Y ¹³ are the same or different hydrogen atoms or optionally substituted C1-C4 alkyl, Z ⁴ to Z ⁶ are the same or different, and —O—, —NY ^14A —, -CO-O -, - O- CO -, - CO-NY 14B -, - NY 14C -CO- or -NY ^14D -CO-0- and is (wherein, Y ^14A ~ Y ^14D are identical or different A hydrogen atom or an optionally substituted C1-C4 alkyl), p ⁴ is an integer of 0 to 5, p ⁵ is an integer of 1 to 5, and p ⁶ is an integer of 0 to 5) And
Or not L ⁷ is ^{^{present, - (CY 15 Y 16)}} p7 -, - (CY 17 Y 18) p8 -Z 7 - (CY 19 Y 20) p9 - or ^{^{_{- (CY 21 Y 22) p10}}} -Z 8 - ^{^{_{(CY 23 Y 24) p11 -Z}}} 9 - (CY 25 Y 26) p12 - ( wherein, Y ¹⁵ ~ Y ²⁶ are the same or different and each is a hydrogen atom or an optionally substituted C1-C4 alkyl, Z ⁷ ~ Z ⁹ are the same or ^{different, -O -, - NY 27A -} , - CO-O -, - O-CO -, - CO-NY 27B -, - NY 27C -CO- or -NY ^27D -CO -0- (wherein Y ^27A to Y ^27D are the same or different hydrogen atom or optionally substituted C1-C4 alkyl), p ⁷ is an integer of 1 to 5, and p ⁸ is An integer from 0 to 5, p ⁹ is an integer from 1 to 5, p ¹⁰ is an integer from 0 to 5, p ¹¹ is an integer from 1 to 5, and p ¹² is an integer from 1 to 5 Is)
B ¹ is

Wherein X ² and X ³ are the same or different C1-C4 alkyl, which may be substituted, or together, a C4-C6 heterocycle which may be substituted with an adjacent nitrogen atom. And X ⁴ is an optionally substituted C1-C4 alkyl, and X ⁵ and X ⁶ are the same or different optionally substituted C1-C4 alkyl, or together are adjacent Forms an optionally substituted C4-C6 heterocycle with a nitrogen atom, X ⁷ is an optionally substituted C1-C4 alkyl, and Y ²⁸ to Y ³⁷ are the same or different and are a hydrogen atom or substituted. May be C1-C4 alkyl, and Z ¹⁰ and Z ¹¹ may be the same or different -O-, -NY ^38A- , -CO-O-, -O-CO-, ^-CO -NY ^38B- , -NY ^38C -CO- or -NY ^38D -CO-0- (wherein Y ^38A to Y ^38D are the same or different and each represents a hydrogen atom or an optionally substituted C1-C4 alkyl). P ¹³ is an integer of 0 to 5, and p ¹⁴ to p ¹⁷ are the same or different and are integers of 1 to 5),
A ² is a pharmaceutically acceptable anion)
Formula (III)

(Where
R ⁷ ~ R ⁹ Are the same or different, linear or branched optionally substituted C8-C24 alkyl, C8-C24 alkenyl or C8-C24 alkynyl,
L ⁸ ~ L ^Ten Are not the same or different, or -Z ¹² -(CY ³⁹ Y ⁴⁰ ) _p18 -Or-Z ¹³ -(CY ⁴¹ Y ⁴² ) _p19 -Z ¹⁴ -(CY ⁴³ Y ⁴⁴ ) _p20 -(Where Y ³⁹ ~ Y ⁴⁴ Are the same or different hydrogen atoms or optionally substituted C1-C4 alkyl, and Z ¹² ~ Z ¹⁴ Are the same or different, -O-, -NY ^45A -, -CO-O-, -O-CO-, -CO-NY ^45B -, -NY ^45C -CO-, -NY ^45D -CO-0- or -CO- (where Y ^45A ~ Y ^45D Are the same or different hydrogen atoms or optionally substituted C1-C4 alkyls), p ¹⁸ Is an integer from 0 to 5 and p ¹⁹ Is an integer from 1 to 5 and p ²⁰ Is an integer from 0 to 5),
L ¹¹ Does not exist or-(CY ⁴⁶ Y ⁴⁷ ) _p21 -、-(CY ⁴⁸ Y ⁴⁹ ) _p22 -Z ¹⁵ -(CY ⁵⁰ Y ⁵¹ ) _p23 -Or- (CY ⁵² Y ⁵³ ) _p24 -Z ¹⁶ -(CY ⁵⁴ Y ⁵⁵ ) _p25 -Z ¹⁷ -(CY ⁵⁶ Y ⁵⁷ ) _p26 -(Where Y ⁴⁶ ~ Y ⁵⁷ Are the same or different hydrogen atoms or optionally substituted C1-C4 alkyl, and Z ¹⁵ ~ Z ¹⁷ Are the same or different, -O-, -NY ^58A -, -CO-O-, -O-CO-, -CO-NY ^58B -, -NY ^58C -CO-, -NY ^58D -CO-0- or -CO- (where Y ^58A ~ Y ^58D Are the same or different hydrogen atoms or optionally substituted C1-C4 alkyls), p ^{twenty one} Is an integer from 1 to 5 and p ^{twenty two} Is an integer from 0 to 5 and p ^{twenty three} Is an integer from 1 to 5 and p ^{twenty four} Is an integer from 0 to 5 and p ^{twenty five} Is an integer from 1 to 5 and p ²⁶ Is an integer from 1 to 5),
L ¹² Does not exist or-(CY ⁵⁹ Y ⁶⁰ ) _p27 -、-(CY ⁶¹ Y ⁶² ) _p28 -Z ¹⁸ -(CY ⁶³ Y ⁶⁴ ) _p29 -Or- (CY ⁶⁵ Y ⁶⁶ ) _p30 -Z ¹⁹ -(CY ⁶⁷ Y ⁶⁸ ) _p31 -Z ²⁰ -(CY ⁶⁹ Y ⁷⁰ ) _p32 -(Where Y ⁵⁹ ~ Y ⁷⁰ Are the same or different hydrogen atoms or optionally substituted C1-C4 alkyl, and Z ¹⁸ ~ Z ²⁰ Are the same or different, -O-, -NY ^71A -, -CO-O-, -O-CO-, -CO-NY ^71B -, -NY ^71C -CO-, -NY ^71D -CO-0- or -CO- (where Y ^71A ~ Y ^71D Are the same or different hydrogen atoms or optionally substituted C1-C4 alkyls), p ²⁷ Is an integer from 1 to 5 and p ²⁸ Is an integer from 0 to 5 and p ²⁹ Is an integer from 0 to 5 and p ³⁰ Is an integer from 0 to 5 and p ³¹ Is an integer from 1 to 5 and p ³² Is an integer from 0 to 5),
J ¹ And J ² Are the same or different CY ⁷² Or N (where Y ⁷² Is a hydrogen atom, hydroxy, optionally substituted C1-C4 alkyl, optionally substituted C1-C4 alkoxy, or optionally substituted C1-C4 acyloxy),
B ² Is

(Where X ⁸ And X ⁹ Are C1-C4 alkyls which may be the same or differently substituted, or together form an optionally substituted C4-C6 heterocycle with an adjacent nitrogen atom, X ^Ten Is an optionally substituted C1-C4 alkyl, and X ¹¹ And X ¹² Are C1-C4 alkyls which may be the same or differently substituted, or together form an optionally substituted C4-C6 heterocycle with an adjacent nitrogen atom, X ¹³ Is optionally substituted C1-C4 alkyl, Y ⁷³ ~ Y ⁸² Are the same or different hydrogen atoms or optionally substituted C1-C4 alkyl, and Z ^{twenty one} And Z ^{twenty two} Are the same or different -O-, -NY ^83A -, -CO-O-, -O-CO-, -CO-NY ^83B -, -NY ^83C -CO- or -NY ^83D -CO-0- (where Y ^83A ~ Y ^83D Are the same or different hydrogen atoms or optionally substituted C1-C4 alkyls), p ³³ Is an integer from 0 to 5 and p ³⁴ ~ P ³⁷ Are the same or different and are integers of 1 to 5),
A ^Three Is a pharmaceutically acceptable anion),
Formula (IV)

(Where
R ¹⁰ to R ¹² are the same or different and may be linear or branched C8-C24 alkyl, C8-C24 alkenyl or C8-C24 alkynyl,
Or L ¹³ is ^{^{absent, -Z 23 - (CY 84 Y}} 85) p38 - or ^{^{^{-Z 24 - (CY 86 Y 87}}} ) p39 -Z 25 - (CY 88 Y 89) p40 - ( wherein, Y ⁸⁴ ~ Y ⁸⁹ is the same or different hydrogen atom or C1-C4 alkyl which may be substituted, Z ²³ to Z ²⁵ are the same or different, and —O—, —NY ^90A —, —CO—O—, — O-CO -, - CO- NY 90B -, - NY 90C -CO- or -NY ^90D -CO-0- and is (wherein, Y ^90A ~ Y ^90D being hydrogenated atom or a substituent identical or different P ³⁸ to p ⁴⁰ are the same or different and are integers of 1 to 5),
L ¹⁴ and L ¹⁵ is absent the same or ^{^{different, -Z 26 - (CY 91 Y}} 92) p41 - or ^{^{^{-Z 27 - (CY 93 Y 94}}} ) p42 -Z 28 - (CY 95 Y 96) p43 - (In the formula, Y ⁹¹ to Y ⁹⁶ are the same or different hydrogen atoms or optionally substituted C1-C4 alkyl, Z ²⁶ to Z ²⁸ are the same or different, and —O—, ^{—NY 97A} —, -CO-O -, - O- CO -, - CO-NY 97B -, - NY 97C -CO -, - during NY ^97D -CO-0- or a -CO- (wherein, Y ^97A ~ Y ^97D is P ⁴¹ is an integer of 0 to 5, p ⁴² is an integer of 1 to 5, and p ⁴³ is an integer of 0 to 5 Is an integer)
Or L ¹⁶ is ^{^{absent, - (CY 98 Y 99)}} p44 -, - (CY 100 Y 101) p45 -Z 29 - (CY 102 Y 103) p46 - or ^{^{_{- (CY 104 Y 105) p47}}} -Z 30 - ^{^{_{(CY 106 Y 107) p48 -Z}}} 31 - (CY 108 Y 109) p49 - ( wherein, Y ⁹⁸ ~ Y ¹⁰⁹ are the same or different and each is a hydrogen atom or an optionally substituted C1-C4 alkyl, Z ²⁹ ~ Z ³¹ are the same or ^{different, -O -, - NY 110A -} , - CO-O -, - O-CO -, - CO-NY 110B -, - NY 110C -CO -, - NY 110D -CO -0- or -CO- (wherein Y ^110A to Y ^110D are the same or different and are a hydrogen atom or an optionally substituted C1-C4 alkyl), and p ⁴⁴ is an integer of 1 to 5 , P ⁴⁵ is an integer from 0 to 5, p ⁴⁶ is an integer from 1 to 5, p ⁴⁷ is an integer from 0 to 5, p ⁴⁸ is an integer from 1 to 5, and p ⁴⁹ is from 1 to Is an integer of 5),
J ³ is CY ¹¹¹ or N (where Y ¹¹¹ is a hydrogen atom, hydroxy, an optionally substituted C1-C4 alkyl, an optionally substituted C1-C4 alkoxy, or an optionally substituted C1- C4 acyloxy),
X ¹⁴ and X ¹⁵ are the same or different optionally substituted C1-C4 alkyl, or together form an optionally substituted C4-C6 heterocycle with an adjacent nitrogen atom;
A ⁴ is a pharmaceutically acceptable anion), or formula (V ′) or formula (V ″)

(Where
R ¹³ to R ¹⁸ are the same or different and are linear or branched C8-C24 alkyl, C8-C24 alkenyl or C8-C24 alkynyl,
Y ¹¹² to Y ¹¹⁵ are the same or different and each represents a hydrogen atom, hydroxy, or optionally substituted C1-C4 alkyl,
Or L ¹⁷ ~ L ¹⁹ and L ²² ~ L ²⁴ are not present in the same or ^{^{different, -Z 32 - (CY 116 Y}} 117) p51 - or ^{^{^{-Z 33 - (CY 118 Y 119}}} ) p52 -Z 34 - (CY ¹²⁰ Y ¹²¹ ) _p53- (wherein Y ¹¹⁶ to Y ¹²¹ are the same or different hydrogen atoms or optionally substituted C1-C4 alkyl, Z ³² to Z ³⁴ are the same or different, and —O— ^{, -NY 122A -, - CO-} O -, - O-CO -, - CO-NY 122B -, - NY 122C -CO -, - NY 122D -CO-0- or a -CO- (wherein, Y ^122A to Y ^122D are the same or different hydrogen atoms or optionally substituted C1-C4 alkyl), p ⁵¹ is an integer of 0 to 5, p ⁵² is an integer of 1 to 5, and p ⁵³ is an integer from 0 to 5),
Or L ²⁰ and L ²⁵ are not present in the same or ^{^{different, - (CY 123 Y 124)}} p54 -, - (CY 125 Y 126) p55 -Z 35 - (CY 127 Y 128) p56 - or - (CY ¹²⁹ Y ^{_{^{130) p57 -Z 36 - (CY}}} 131 Y 132) p58 -Z 37 - (CY 133 Y 134) p59 - ( wherein, Y ¹²³ ~ Y ¹³⁴ may be a hydrogen atom or a substituent identical or different C1 an -C4 alkyl, Z ³⁵ ~ Z ³⁷ are the same or different ^{and, -O -, - NY 135A -} , - CO-O -, - O-CO -, - CO-NY 135B -, - NY 135C -CO -, ^{-NY 135D} -CO-0- or -CO- (wherein Y ^135A to Y ^135D are the same or different and are a hydrogen atom or an optionally substituted C1-C4 alkyl), and p ⁵⁴ is An integer from 1 to 5, p ⁵⁵ is an integer from 0 to 5, p ⁵⁶ is an integer from 1 to 5, p ⁵⁷ is an integer from 0 to 5, and p ⁵⁸ is an integer from 1 to 5 And p ⁵⁹ is an integer from 1 to 5),
Or L ²¹ and L ²⁶ are not present in the same or ^{^{different, - (CY 136 Y 137)}} p60 -, - (CY 138 Y 139) p61 -Z 38 - (CY 140 Y 141) p62 - or - (CY ¹⁴² Y ^{_{^{143) p63 -Z 39 - (CY}}} 144 Y 145) p64 -Z 40 - (CY 146 Y 147) p65 - ( wherein, Y ¹³⁶ ~ Y ¹⁴⁷ may be a hydrogen atom or a substituent identical or different C1 an -C4 alkyl, Z ³⁸ ~ Z ⁴⁰ are the same or ^{different, -O -, - NY 148A -} , - CO-O -, - O-CO -, - CO-NY 148B -, - NR 148C -CO -, ^{-NY 148D} -CO-0- or -CO- (wherein Y ^148A to Y ^148D are the same or different hydrogen atom or optionally substituted C1-C4 alkyl), and p ⁶⁰ is An integer from 1 to 5, p ⁶¹ is an integer from 0 to 5, p ⁶² is an integer from 0 to 5, p ⁶³ is an integer from 0 to 5, and p ⁶⁴ is an integer from 1 to 5 And p ⁶⁵ is an integer from 0 to 5),
B ³ and B ⁴ are the same or different,

Wherein X ¹⁶ and X ¹⁷ are the same or different C1-C4 alkyl, which may be substituted, or together, a C4-C6 heterocycle which may be substituted with an adjacent nitrogen atom. And X ¹⁸ is an optionally substituted C1-C4 alkyl, and X ¹⁹ and X ²⁰ are the same or different optionally substituted C1-C4 alkyl, or together are adjacent Forms an optionally substituted C4-C6 heterocycle with a nitrogen atom, X ²¹ is an optionally substituted C1-C4 alkyl, and Y ¹⁴⁹ to Y ¹⁵⁸ are the same or different and are a hydrogen atom or substituted May be C1-C4 alkyl, and Z ⁴¹ and Z ⁴² may be the same or different, -O-, ^{-NY 159A-} , ^-CO -O-, -O- ^CO- , ^-CO -NY ^159B- , ^-NY ^159C -CO- or -NY ^159D -CO-0- and is (wherein, Y ^159A ~ Y ^159D are the same or different and each a hydrogen atom or an optionally substituted C1-C4 a A kill), p ⁶⁶ represents an integer of ^{0 ~ 5, p 67 ~ p} 70 are the same or different and is an integer of 1-5),
A ⁵ and A ⁶ are the same or different pharmaceutically acceptable anions)
The compound represented by these is mentioned.

Hereinafter, the compounds represented by formulas (I) to (IV), (V ′) and (V ″) are also referred to as compounds (I) to (IV), (V ′) and (V ″), respectively. is there. The same applies to the compounds of other formula numbers.

The definition of each group of formulas (I) to (V ″) will be described below.
Examples of linear or branched C8-C24 alkyl include octyl, nonyl, decyl, undecyl, dodecyl, tridecyl, tetradecyl, 2,6,10-trimethylundecyl, pentadecyl, 3,7,11-trimethyldodecyl, Hexadecyl, heptadecyl, octadecyl, 6,10,14-trimethylpentadecan-2-yl, nonadecyl, 2,6,10,14-tetramethylpentadecyl, icosyl, 3,7,11,15-tetramethylhexadecyl, henicosyl , Docosyl, tricosyl, tetracosyl and the like, preferably nonyl, decyl, undecyl, dodecyl, tridecyl, tetradecyl, pentadecyl, hexadecyl, heptadecyl, octadecyl, etc., more preferably undecyl, dodecyl, tridecyl, tetradecyl, pentadecyl, Hexadecyl etc. are mentioned.

Examples of linear or branched C9-C18 alkyl include nonyl, decyl, undecyl, dodecyl, tridecyl, tetradecyl, 2,6,10-trimethylundecyl, pentadecyl, 3,7,11-trimethyldodecyl, hexadecyl, Heptadecyl, octadecyl, 6,10,14-trimethylpentadecan-2-yl, and the like, preferably nonyl, decyl, undecyl, dodecyl, tridecyl, tetradecyl, pentadecyl, hexadecyl, heptadecyl, octadecyl, and the like, more preferably Examples include undecyl, dodecyl, tridecyl, tetradecyl, pentadecyl, hexadecyl and the like.

The linear or branched C8-C24 alkenyl may be a linear or branched C8-24 alkenyl containing 1 to 3 double bonds, such as (Z) -tridec-8-enyl, (Z) -tetradec-9-enyl, (Z) -pentadeca-8-enyl, (Z) -hexadeca-9-enyl, (Z) -heptadeca-5-enyl, (Z) -octadeca-6-enyl, (Z) -heptadeca-8-enyl, (Z) -octadeca-9-enyl, (E) -heptadeca-8-enyl, (E) -octadeca-9-enyl, (Z) -heptadeca-10-enyl, (Z) -octadeca-11-enyl, (8Z, 11Z) -heptadeca-8,11-dienyl, (9Z, 12Z) -octadeca-9,12-dienyl, (8Z, 11Z, 14Z) -octadeca-8, 11,14-trienyl, (9Z, 12Z, 15Z) -octadeca-9,12,15-trienyl, (Z) -nonadec-10-enyl, (Z) -icosa-11-enyl, (10Z, 13Z)- Nonadeca-10,13-dienyl, (11Z, 14Z) -icosa-11,14-dienyl, 2,6,10-trimethylundeca-1,5,9-trienyl, 3,7,11 -Trimethyldodeca-2,6,10-trienyl, 2,6,10,14-tetramethylpentadec-1-enyl, 3,7,11,15-tetramethylhexadec-2-enyl, etc. Preferably (Z) -pentadeca-8-enyl, (Z) -hexadeca-9-enyl, (Z) -heptadeca-5-enyl, (Z) -octadeca-6-enyl, (Z) -heptadeca-8- Enil, (Z) -octadec-9-enyl, (8Z, 11Z) -heptadeca-8,11-dienyl, (9Z, 12Z) -octadeca-9,12-dienyl, and more preferably (Z) -Heptadeca-8-enyl, (Z) -octadeca-9-enyl, (8Z, 11Z) -heptadeca-8,11-dienyl, (9Z, 12Z) -octadeca-9,12-dienyl and the like.

Linear or branched C15-C20 alkenyl includes (Z) -pentadeca-8-enyl, (Z) -hexadeca-9-enyl, (Z) -heptadeca-5-enyl, (Z) -octadeca- 6-enyl, (Z) -heptadeca-8-enyl, (Z) -octadeca-9-enyl, (E) -heptadeca-8-enyl, (E) -octadeca-9-enyl, (Z) -heptadeca- 10-enyl, (Z) -octadeca-11-enyl, (8Z, 11Z) -heptadeca-8,11-dienyl, (9Z, 12Z) -octadeca-9,12-dienyl, (8Z, 11Z, 14Z)- Octadeca-8,11,14-trienyl, (9Z, 12Z, 15Z) -octadeca-9,12,15-trienyl, (Z) -nonadec-10-enyl, (Z) -icosa-11-enyl, (10Z , 13Z) -nonadeca-10,13-dienyl, (11Z, 14Z) -icosa-11,14-dienyl, 3,7,11,15-tetramethylhexadec-2-enyl, etc., preferably ( Z) -pentadec-8-enyl, (Z) -hexadec-9-enyl, (Z) -heptadeca-5-enyl, (Z) -octadeca-6-enyl, (Z) -hepta And more preferred are -8-enyl, (Z) -octadeca-9-enyl, (8Z, 11Z) -heptadeca-8,11-dienyl, (9Z, 12Z) -octadeca-9,12-dienyl, and the like. (Z) -heptadeca-8-enyl, (Z) -octadeca-9-enyl, (8Z, 11Z) -heptadeca-8,11-dienyl, (9Z, 12Z) -octadeca-9,12-dienyl, etc. Can be mentioned.

In the present invention, C8-C24 alkenyl also includes a group having a cyclopropane ring in which a methylene biradical is added formally to a linear bond or branched C8-C24 alkenyl double bond. Is done. For example, a group having the following cyclopropane ring corresponding to (Z) -hexadec-9-enyl, (9Z, 12Z) -octadeca-9,12-dienyl, (8Z, 11Z) -heptadeca-8,11-dienyl

and

Etc.

The linear or branched C8-C24 alkynyl may be a linear or branched C8-24 alkynyl containing 1 to 3 triple bonds, for example, dodeca-11-ynyl, trideca-12- Inyl, pentadec-6-ynyl, hexadec-7-ynyl, pentadec-4,6-diynyl, hexadec-5,7-diynyl, heptadec-8-ynyl, octadec-9-ynyl, etc., preferably pentadec- 6-Inyl, hexadec-7-ynyl, pentadec-4,6-diynyl, hexadec-5,7-diynyl, heptadec-8-ynyl, octadec-9-ynyl, and the like, more preferably heptadeca-8-inyl And octadec-9-ynyl.

Examples of C1-C4 alkyl include methyl, ethyl, propyl, isopropyl, cyclopropyl, butyl, isobutyl, sec-butyl, tert-butyl, cyclobutyl, cyclopropylmethyl and the like, preferably methyl, ethyl and the like. More preferably, methyl is mentioned.

The alkyl part of C1-C4 alkoxy which may be substituted has the same meaning as the C1-C4 alkyl.

Examples of the substituent in linear or branched C8-C24 alkyl, C8-C24 alkenyl or C8-C24 alkynyl which may be substituted include hydroxy, alkoxy, alkoxycarbonyl, nitro, cyano, fluoro, chloro, bromo, etc. Is mentioned. Of these substituents, the alkyl moiety in alkoxy and alkoxycarbonyl has the same meaning as the C1-C4 alkyl.

Examples of the substituent in the optionally substituted C1-C4 alkyl include amino, monoalkylamino, dialkylamino, pyrrolidin-2-yl, pyrrolidin-3-yl, piperidin-2-yl, piperidin-3-yl, and piperidine 4-yl, morpholin-2-yl, morpholin-3-yl, hydroxy, alkoxy, alkoxycarbonyl, hydroxycarbonyl, carbamoyl, monoalkylcarbamoyl, dialkylcarbamoyl, nitro, cyano, fluoro, chloro, bromo and the like. Of these substituents, the alkyl moiety in monoalkylamino, dialkylamino, alkoxy, alkoxycarbonyl, monoalkylcarbamoyl and dialkylcarbamoyl has the same meaning as the C1-C4 alkyl. Two alkyls in dialkylamino and dialkylcarbamoyl may be the same or different.

In the present invention, pyrrolidin-2-yl, pyrrolidin-3-yl, piperidin-2-yl, piperidin-3-yl, piperidin-4-yl, morpholin-2-yl and morpholin-3-yl are each a ring. Includes those in which a C1-C3 alkyl group such as methyl or ethyl is bonded to the nitrogen atom.
Examples of C1-C3 alkyl include methyl, ethyl, propyl, isopropyl, cyclopropyl and the like, preferably methyl, ethyl and the like, more preferably methyl and the like.

Examples of the C4-C6 heterocycle formed by X ² and X ³ together with the adjacent nitrogen atom include pyrrolidine, piperidine, morpholine, azepan and the like, preferably pyrrolidine, piperidine and the like. Examples of the substituent in the optionally substituted C4-C6 heterocycle formed by X ² and X ³ together with the adjacent nitrogen atom include optionally substituted C1-C4 alkyl (as defined above), amino Monoalkylamino, dialkylamino, pyrrolidin-2-yl, pyrrolidin-3-yl, piperidin-2-yl, piperidin-3-yl, piperidin-4-yl, morpholin-2-yl, morpholin-3-yl, Examples include hydroxy, alkoxy, alkoxycarbonyl, hydroxycarbonyl, carbamoyl, monoalkylcarbamoyl, dialkylcarbamoyl, nitro, cyano, fluoro, chloro, bromo and the like. Of these substituents, the alkyl moiety in monoalkylamino, dialkylamino, alkoxy, alkoxycarbonyl, monoalkylcarbamoyl and dialkylcarbamoyl has the same meaning as the C1-C4 alkyl. Two alkyls in dialkylamino and dialkylcarbamoyl may be the same or different.

The heterocyclic portion and the substituent portion of the optionally substituted C4-C6 heterocycle formed by X ⁵ and X ⁶ together with the adjacent nitrogen atom are as defined above.

The heterocyclic portion and the substituent portion of the optionally substituted C4-C6 heterocycle formed by X ⁸ and X ⁹ together with the adjacent nitrogen atom are as defined above.

The heterocyclic portion and the substituent portion of the optionally substituted C4-C6 heterocyclic ring formed by X ¹¹ and X ¹² together with the adjacent nitrogen atom are as defined above.

The heterocyclic portion and the substituent portion of the optionally substituted C4-C6 heterocyclic ring formed by X ¹⁴ and X ¹⁵ together with the adjacent nitrogen atom are as defined above.

The heterocyclic portion and the substituent portion of the optionally substituted C4-C6 heterocycle formed by X ¹⁶ and X ¹⁷ together with the adjacent nitrogen atom are as defined above.

The heterocyclic portion and the substituent portion of the optionally substituted C4-C6 heterocyclic ring formed by X ¹⁹ and X ²⁰ together with the adjacent nitrogen atom are as defined above.

Examples of the acyl in C1-C4 acyloxy include formyl, acetyl, propanoyl, 2-methylpropanoyl, cyclopropanoyl, butanoyl, and preferably acetyl and the like.

Examples of the substituent in C1-C4 acyloxy which may be substituted include amino, monoalkylamino, dialkylamino, pyrrolidin-2-yl, pyrrolidin-3-yl, piperidin-2-yl, piperidin-3-yl and piperidine 4-yl, morpholin-2-yl, morpholin-3-yl, hydroxy, alkoxy, alkoxycarbonyl, hydroxycarbonyl, carbamoyl, monoalkylcarbamoyl, dialkylcarbamoyl, nitro, cyano, fluoro, chloro, bromo and the like. Of these substituents, the alkyl moiety in monoalkylamino, dialkylamino, alkoxy, alkoxycarbonyl, monoalkylcarbamoyl and dialkylcarbamoyl has the same meaning as the C1-C4 alkyl. Two alkyls in dialkylamino and dialkylcarbamoyl may be the same or different.

The quaternary ammonium group means a group having a nitrogen atom having four covalent bonds with four carbon atoms. The quaternary ammonium group is always positively charged regardless of the surrounding pH, unlike a hydrogen atom added to a primary to tertiary amine.

Examples of pharmaceutically acceptable anions include chloride ions, bromide ions, iodide ions, nitrate ions, sulfate ions, phosphate ions and other inorganic ions, acetate ions, oxalate ions, maleate ions, fumarate. Examples include, but are not limited to, organic acid ions such as acid ions, citrate ions, benzoate ions, and methanesulfonate ions.

In the formula (I), R ¹ to R ³ are preferably the same linear or branched C8-C24 alkyl, C8-C24 alkenyl or C8-C24 alkynyl, and the same linear or branched Are more preferably C8-C24 alkyl or C8-C24 alkenyl, and are the same linear or branched C15-C20 alkenyl, or the same linear or branched C9-C18 alkyl. More preferably, it is the same linear C15-C20 alkenyl, or most preferably the same linear C9-C18 alkyl.

L ¹ to L ³ are the same or different and do not exist, -Z ^1- (CY ¹ Y ² ) _p1 -or -Z ^2- (CY ³ Y ⁴ ) _p2 -Z ^3- (CY ⁵ Y ⁶ ) _p3 -And preferably -Z ^1- (CY ¹ Y ² ) _p1- . Y ¹ to Y ⁶ are the same or different and each represents a hydrogen atom or an optionally substituted C1-C4 alkyl, and Y ¹ to Y ⁶ are preferably a hydrogen atom. Z ¹ ~ Z ³ are the same or ^{different, -O -, - NY 7A -} , - CO-O -, - O-CO -, - CO-NY 7B -, - NY 7C -CO- or -NY ^7D - CO-0-, -O-, -CO-O-, -O-CO-, -CO-NY ^7B- , and -NY ^7C -CO- are preferred. Y ^7A to Y ^7D are the same or different and are a hydrogen atom or an optionally substituted C1-C4 alkyl, preferably a hydrogen atom or methyl. p ¹ to p ³ are the same or different and are integers of 1 to 5, preferably 1 or 2.

L ¹ to L ³ may be the same or different from -O- (CY ¹ Y ² ) _p1- , -CO-O- (CY ¹ Y ² ) _p1- , -O-CO- (CY ¹ Y ² ) _p1 -, -CO-NY ^7B- (CY ¹ Y ² ) _p1 -or -NY ^7C -CO- (CY ¹ Y ² ) _p1- , preferably the same or different -CO-O- (CY ¹ Y ² ) _p1 -or -O-CO- (CY ¹ Y ² ) _p1- is more preferable, and -CO-O- (CH ₂ ) _2- is more preferable.

In formula (I), at ^{least one} of L ¹ to L ³ is the same or different and is —CO—O— (CY ¹ Y ² ) _p1 — or —O—CO— (CY ¹ Y ² ) _p1 — R ¹ to R ³ are preferably the same linear C15-C20 alkenyl or the same linear C9-C18 alkyl.

At least one of L ¹ to L ³ does not exist or is -O- (CY ¹ Y ² ) _p1- , -O-CO- (CY ¹ Y ² ) _p1 -or -NY ^7C -CO- (CY ¹ Y ² ) when _p1- , a positively charged nitrogen atom (N ⁺ ), -O- (CY ¹ Y ² ) _p1- , -O-CO- (CY ¹ Y ² ) _p1 -or -NR ⁶ R ¹ to R ³ bonded to —CO— (CY ¹ Y ² ) _p1 — are the same or different, and octyl, decyl, dodecyl, tetradecyl, hexadecyl, octadecyl, icosyl, docosyl, tetracosyl, (Z) -tetradeca- 9-enyl, (Z) -hexadeca-9-enyl, (Z) -octadeca-6-enyl, (Z) -octadeca-9-enyl, (E) -octadec-9-enyl, (Z) -octadeca- 11-enyl, (9Z, 12Z) -octadeca-9,12-dienyl, (9Z, 12Z, 15Z) -octadeca-9,12,15-trienyl, (Z) -icosa-11-enyl, (11Z, 14Z ) -Icosa-11,14-dienyl, 3,7,11-trimethyldodeca-2,6,10-trienyl, 3,7,11,15-tetramethylhexadec-2-enyl, etc. And more preferably, dodecyl, tetradecyl, hexadecyl, (Z) -hexadec-9-enyl, (Z) -octadec-6-enyl, (Z) -octadec-9-enyl, (9Z, 12Z) -octadeca More preferred is -9,12-dienyl.

When at least one of L ¹ to L ³ is -CO-O- (CY ¹ Y ² ) _p1 -or -CO-NY ^7B- (CY ¹ Y ² ) _p1- , -CO-O- ( R ¹ to R ³ bonded to CY ¹ Y ² ) _p1 -or -CO-NY ^7B- (CY ¹ Y ² ) _p1- are the same or different and are each nonyl, undecyl, tridecyl, pentadecyl, heptadecyl, nonadecyl, Henicosyl, tricosyl, (Z) -tridec-8-enyl, (Z) -pentadeca-8-enyl, (Z) -heptadeca-5-enyl, (Z) -heptadeca-8-enyl, (E) -heptadeca- 8-enyl, (Z) -heptadeca-10-enyl, (8Z, 11Z) -heptadeca-8,11-dienyl, (8Z, 11Z, 14Z) -octadeca-8,11,14-trienyl, (Z)- Nonadeca-10-enyl, (10Z, 13Z) -Nonadeca-10,13-dienyl, (11Z, 14Z) -icosa-11,14-dienyl, 2,6,10-trimethylundeca-1,5,9- More preferred are trienyl, 2,6,10,14-tetramethylpentadec-1-enyl and the like, undecyl, tridecyl , Pentadecyl, (Z) -pentadeca-8-enyl, (Z) -heptadeca-5-enyl, (Z) -heptadeca-8-enyl, (8Z, 11Z) -heptadeca-8,11-dienyl, etc. More preferably.

X ¹ is preferably methyl, hydroxypropyl or hydroxyethyl, more preferably methyl.

In the formula (II), R ⁴ to R ⁶ are preferably the same linear or branched C8-C24 alkyl, C8-C24 alkenyl or C8-C24 alkynyl, and the same linear or branched Are more preferably C8-C24 alkyl or C8-C24 alkenyl, and are the same linear or branched C15-C20 alkenyl, or the same linear or branched C9-C18 alkyl. More preferably, it is the same linear C15-C20 alkenyl, or most preferably the same linear C9-C18 alkyl.

L ⁴ ~ L ⁶ is absent same or ^{^{different, -Z 4 - (CY 8 Y}} 9) p4 - or ^{^{^{-Z 5 - (CY 10 Y 11}}} ) p5 -Z 6 - (CY 12 Y 13) p6 - ^{^{and, -Z 4 - (CY 8 Y}} 9) p4 - or ^{^{^{-Z 5 - (CY 10 Y 11}}} ) p5 -Z 6 - (CY 12 Y 13) p6 - is preferably, -Z ⁴ - More preferably, (CY ⁸ Y ⁹ ) _p4- . Y ⁸ to Y ¹³ are the same or different and each represents a hydrogen atom or optionally substituted C1-C4 alkyl, and Y ⁸ to Y ¹³ are preferably hydrogen atoms. Z ⁴ to Z ⁶ may be the same or different, -O-, -NY ^14A- , -CO-O-, -O-CO-, ^-CO -NY ^14B- , -NY ^14C -CO- or -NY ^14D- CO-0-, -O-, -CO-O-, -O-CO-, -CO-NY ^14B- , -NY ^14C -CO- are preferred. Y ^27A to Y ^27D are the same or different and each represents a hydrogen atom or an optionally substituted C1-C4 alkyl, preferably a hydrogen atom or methyl. p ⁴ is an integer of 0 to 5, p ⁵ is an integer of 1 to 5, and p ⁶ is an integer of 0 to 5, both of which are preferably 1 or 2.

L ⁴ to L ⁶ may be the same or different from -O- (CY ⁸ Y ⁹ ) _p4- , -CO-O- (CY ⁸ Y ⁹ ) _p4- , -O-CO- (CY ⁸ Y ⁹ ) _p4 ^{-, - CO-NY 14B -} (CY 8 Y 9) p4 -, - NY 14C -CO- (CY 8 Y 9) p4 -, - NY 14D -CO-O- (CY 8 Y 9) p4 - or - O-CO- (CY ¹⁰ Y ¹¹ ) _p5 -Z ^6- (CY ¹² Y ¹³ ) _p6- is preferred, and the same or different -CO-O- (CY ⁸ Y ⁹ ) _p4- , -O- More preferably, CO- (CY ⁸ Y ⁹ ) _p4 -or -O-CO- (CY ¹⁰ Y ¹¹ ) _p5 -O- (CY ¹² Y ¹³ ) _p6- , identically -CO-O-CH ₂ -Is more preferable.

In the formula (II), one or more of L ₄ to L ₆ are the same or different, and —CO—O— (CY ⁸ Y ⁹ ) _p4 −, —O—CO— (CY ⁸ Y ⁹ ) _p4 —, or -O-CO- (CY ¹⁰ Y ¹¹ ) _p5 -O- (CY ¹² Y ¹³ ) _p6- , and R ⁴ to R ⁶ are the same linear C15-C20 alkenyl, or the same linear C9-C18 alkyl is preferable.

At least one of L ⁴ to L ⁶ does not exist or is -O- (CY ⁸ Y ⁹ ) _p4- , -O-CO- (CY ⁸ Y ⁹ ) _p4- , -NY ^14C -CO- (CY ⁸ Y ^{_{^{9) p4 -, - NY 14D}}} -CO-0- or ^{^{-O-CO- (CY 10 Y 11}} ) p5 - (CY 12 Y 13) p6 - a if the carbon atom adjacent to L ^7, - O- (CY ⁸ Y ⁹ ) _p4- , -O-CO- (CY ⁸ Y ⁹ ) _p4- , -NY ^14C -CO- (CY ⁸ Y ⁹ ) _p4- , -NY ^14D -CO-0- or- R ⁷ to R ⁹ bonded to O-CO- (CY ¹⁰ Y ¹¹ ) _p5 -Z ^6- (CY ¹² Y ¹³ ) _p6- are the same or different and are each octyl, decyl, dodecyl, tetradecyl, hexadecyl, octadecyl , Icosyl, docosyl, tetracosyl, (Z) -tetradec-9-enyl, (Z) -hexadeca-9-enyl, (Z) -octadeca-6-enyl, (Z) -octadeca-9-enyl, (E) -Octadeca-9-enyl, (Z) -octadeca-11-enyl, (9Z, 12Z) -octadeca-9,12-dienyl, (9Z, 12Z, 15Z) -octadeca-9,12,15-trienyl, ( Z) -icosa-11-enyl, (11Z, 14Z) -icosa-11,14-di Nyl, 3,7,11-trimethyldodeca-2,6,10-trienyl, 3,7,11,15-tetramethylhexadec-2-enyl and the like, dodecyl, tetradecyl, hexadecyl, (Z More preferred are) -hexadeca-9-enyl, (Z) -octadeca-6-enyl, (Z) -octadeca-9-enyl, (9Z, 12Z) -octadeca-9,12-dienyl, and the like.

When at least one of L ⁴ to L ⁶ is -CO-O- (CY ⁸ Y ⁹ ) _p4 -or -CO-NY ^14B- (CY ⁸ Y ⁹ ) _p4- , -CO-O- ( R ⁴ to R ⁶ bonded to CY ⁸ Y ⁹ ) _p4 -or -CO-NY ^14B- (CY ⁸ Y ⁹ ) _p4- are the same or different and are each nonyl, undecyl, tridecyl, pentadecyl, heptadecyl, nonadecyl, Henicosyl, tricosyl, (Z) -tridec-8-enyl, (Z) -pentadeca-8-enyl, (Z) -heptadeca-5-enyl, (Z) -heptadeca-8-enyl, (E) -heptadeca- 8-enyl, (Z) -heptadeca-10-enyl, (8Z, 11Z) -heptadeca-8,11-dienyl, (8Z, 11Z, 14Z) -octadeca-8,11,14-trienyl, (Z)- Nonadeca-10-enyl, (10Z, 13Z) -Nonadeca-10,13-dienyl, (11Z, 14Z) -icosa-11,14-dienyl, 2,6,10-trimethylundeca-1,5,9- Preferably, trienyl, 2,6,10,14-tetramethylpentadec-1-enyl, etc., undecyl, tridecyl It may be pentadecyl, (Z) -pentadec-8-enyl, (Z) -heptadeca-5-enyl, (Z) -heptadeca-8-enyl, (8Z, 11Z) -heptadeca-8,11-dienyl, etc. Further preferred.

L ⁷ is not present,-(CY ¹⁵ Y ¹⁶ ) _p7 -,-(CY ¹⁷ Y ¹⁸ ) _p8 -O-CO- (CY ¹⁹ Y ²⁰ ) _p9-or- (CY ¹⁷ Y ¹⁸ ) _p8 -NY ^{27C 2} —CO— (CY ¹⁹ Y ²⁰ ) _p9 — is preferable, and it is not present, or more preferably — (CY ¹⁵ Y ¹⁶ ) _p7 —. In this case, B ¹ is

And is more preferably —N ⁺ (CH ₃ ) ₃ .

When L ⁷ is — (CY ¹⁵ Y ¹⁶ ) _p7 —, p ⁷ is preferably 1 to 3, more preferably 1 to 2, still more preferably 1, and Y ¹⁵ to Y ¹⁶ is preferably each a hydrogen atom. B ¹ is preferably —N ⁺ (CH ₃ ) ₃ .

When L ⁷ _is- (CY ¹⁷ Y ¹⁸ ) _p8 -O-CO- (CY ¹⁹ Y ²⁰ ) _p9-or- (CY ¹⁷ Y ¹⁸ ) _p8 -NY ^27C -CO- (CY ¹⁹ Y ²⁰ ) _p9- , P ⁸ is 0 to 3, p ⁹ is preferably 1 to 3, p ⁸ is 0 to 1, more preferably p ⁹ is 1 to 3, and Y ¹⁷ to Y ²⁰ are Each is a hydrogen atom, and Y ^27C is preferably a hydrogen atom or methyl. B ¹ is preferably —N ⁺ (CH ₃ ) ₃ .

X ² and X ³ are preferably the same or different and are each methyl or ethyl, or preferably taken together to form an optionally substituted C4-C6 heterocycle with adjacent nitrogen atoms. More preferably, they together or together with the adjacent nitrogen atom form pyrrolidine or piperidine, more preferably methyl.

X ⁴ is preferably methyl, ethyl, hydroxypropyl or hydroxyethyl, and more preferably methyl.
X ² and X ³ are the same or different and are each methyl or ethyl, X ⁴ is preferably methyl, ethyl, hydroxypropyl, hydroxyethyl or the like, and X ² to X ⁴ are preferably methyl. preferable.

B ¹ is

L ⁷ is not present or-(CY ¹⁵ Y ¹⁶ ) _p7 -,-(CY ¹⁷ Y ¹⁸ ) _p8 -O-CO- (CY ¹⁹ Y ²⁰ ) _p9-or- (CY ¹⁷ Y ¹⁸ ) _p8 it is one preferred embodiment of the present invention is a ^{^{- -NY 27C -CO- (CY 19 Y}} 20) p9. In this case, B ¹ is

L ⁷ is not present, -NH-CO- (CH ₂ ) _p9- , -O-CO- (CH ₂ ) _p9- , -CH ₂ -NH-CO- (CH ₂ ) _p9-or- More preferably, it is CH ₂ —O—CO— (CH ₂ ) _p9 —.

In the formula (III), R ⁷ is preferably linear or branched C8-C24 alkyl or C8-C24 alkenyl, linear or branched C15-C20 alkenyl, or linear Alternatively, it is more preferably a branched C9-C18 alkyl, and most preferably the same linear C15-C20 alkenyl, or the same linear C9-C18 alkyl. R ⁸ and R ⁹ are linear or branched C8-C24 alkyl, C8-C24 alkenyl or C8-C24 alkynyl, and are preferably the same, linear or branched C15-C20 alkenyl. Or a linear or branched C9-C18 alkyl and more preferably the same, a linear C15-C20 alkenyl, or a linear C9-C18 alkyl. Most preferably, they are identical.

L ⁸ is ^{^{absent, -Z 12 - (CY 39 Y}} 40) p18 - or ^{^{^{-Z 13 - (CY 41 Y 42}}} ) p19 -Z 14 - (CY 43 Y 44) p20 - a is absent or ^{^{^{-Z 12 - (CY 39 Y 40}}} ) p18 - is preferably. L ⁹ and L ¹⁰ is absent same or ^{^{different, -Z 12 - (CY 39 Y}} 40) p18 - or ^{^{^{-Z 13 - (CY 41 Y 42}}} ) p19 -Z 14 - (CY 43 Y 44) p20 - and, the same or different and either missing or ^{^{^{-Z 12 - (CY 39 Y 40}}} ) p18 - is preferably. Y ³⁹ to Y ⁴⁴ are the same or different and each represents a hydrogen atom or optionally substituted C1-C4 alkyl, and Y ³⁹ to Y ⁴⁴ are preferably hydrogen atoms. Z ¹² ~ Z ¹⁴ are the same or ^{different, -O -, - NY 45A -} , - CO-O -, - O-CO -, - CO-NY 45B -, - NY 45C -CO -, - NY 45D - CO-0- or a -CO-, -CO-O -, - O-CO -, - CO-NY 45B -, - it is preferable NY ^45C -CO- or -CO-. Y ^45A to Y ^45D are the same or different and each represents a hydrogen atom or an optionally substituted C1-C4 alkyl, and preferably a hydrogen atom or methyl. p ¹⁸ is an integer of 0 to 5, and is preferably 0 or 1. p ¹⁹ is an integer of 1 to 5, and is preferably 1 or 2. p ²⁰ is an integer of 0 to 5, and is preferably 0 or 1.

One of L ⁸ to L ¹⁰ is -CO-O- (CY ³⁹ Y ⁴⁰ ) _p18 -or -O-CO- (CY ³⁹ Y ⁴⁰ ) _p18- or two or more of L ⁸ to L ¹⁰ Are the same or different and are -CO-O- (CY ³⁹ Y ⁴⁰ ) _p18 or -O-CO- (CY ³⁹ Y ⁴⁰ ) _p18- , and R ⁷ to R ⁹ are linear C15-C20 alkenyl or C9- C18 alkyl is preferred, and R ⁸ to R ⁹ are preferably the same.

L ⁸ does not exist or is -CO-O- (CY ³⁹ Y ⁴⁰ ) _p18- , _-O -CO- (CY ³⁹ Y ⁴⁰ ) _p18- , -CO-NY ^45B- (CY ³⁹ Y ⁴⁰ ) _p18 -Or -NY ^45C -CO- (CY ³⁹ Y ⁴⁰ ) _p18- is preferred and is absent or -CO-O- (CY ³⁹ Y ⁴⁰ ) _p18- , _-O -CO- (CY ³⁹ Y ⁴⁰ ) _p18 -or -CO-NY ^45B- (CY ³⁹ Y ⁴⁰ ) _p18- , more preferably absent or -CO-O- (CH ₂ ) _p18- , -O-CO- (CH ₂ ) _{pi 8} - or _{_{-CO-NH- (CH 2) p18}} - and still more preferably a.
L ⁹ and L ¹⁰ are the same or different and do not exist, -CO-O- (CY ³⁹ Y ⁴⁰ ) _p18- , _-O -CO- (CY ³⁹ Y ⁴⁰ ) _p18- , -CO-NY ^45B- (CY ³⁹ Y ⁴⁰ ) _p18 -or -NY ^45C -CO- (CY ³⁹ Y ⁴⁰ ) _p18- is preferred and does not exist the same or different, or -CO-O- (CY ³⁹ Y ⁴⁰ ) _p18- Or -O-CO- (CY ³⁹ Y ⁴⁰ ) _p18- , more preferably the same or different, or more preferably -CO-O- (CH ₂ ) _p18- , and the same. Or -CO-O- (CH ₂ ) _p18- is most preferred.

In Formula (III), one of L ⁸ to L ¹⁰ does not exist, or —CO—O— (CY ³⁹ Y ⁴⁰ ) _p18 −, _—O —CO— (CY ³⁹ Y ⁴⁰ ) _p18 —, —CO— NY ^45B- (CY ³⁹ Y ⁴⁰ ) _p18 -or -NY ^45C -CO- (CY ³⁹ Y ⁴⁰ ) _p18- , or two or more of L ⁸ to L ¹⁰ are not the same or different, or- CO-O- (CY ³⁹ Y ⁴⁰ ) _p18- , _-O -CO- (CY ³⁹ Y ⁴⁰ ) _p18- , _-CO -NY ^45B- (CY ³⁹ Y ⁴⁰ ) _p18 -or -NY ^45C -CO- (CY ³⁹ Y ⁴⁰ ) _p18- and R ⁷ to R ⁹ are preferably straight-chain C15-C20 alkenyl or C9-C18 alkyl, and R ⁸ to R ⁹ are preferably the same.

At least one of L ⁸ to L ¹⁰ is not present, or -O- (CY ³⁹ Y ⁴⁰ ) _p18- , -O-CO- (CY ³⁹ Y ⁴⁰ ) _p18- , ^{-NY 45C} -CO- (CY ³⁹ Y ⁴⁰ ) _p18 -or -NY ^45D -CO-0- (CY ³⁹ Y ⁴⁰ ) If _p18- , J ¹ or J ² , -O- (CY ³⁹ Y ⁴⁰ ) _p18- , -O-CO- ^{^{_{(CY 39 Y 40) p18 -}}} , - NY 45C -CO- (CY 39 Y 40) p18 - or ^{^{-NY 45D -CO-0- (CY 39}} Y 40) p18 - binding to R ⁷ ~ R ⁹ is Same or different, octyl, decyl, dodecyl, tetradecyl, hexadecyl, octadecyl, icosyl, docosyl, tetracosyl, (Z) -tetradec-9-enyl, (Z) -hexadec-9-enyl, (Z) -octadeca- 6-enyl, (Z) -octadeca-9-enyl, (E) -octadeca-9-enyl, (Z) -octadeca-11-enyl, (9Z, 12Z) -octadec-9,12-dienyl, (9Z , 12Z, 15Z) -Octadeca-9,12,15-trienyl, (Z) -icosa-11-enyl, (11Z, 14Z) -icosa-11,14-dienyl, 3,7,11-trimethyldodeca-2 , 6,10 -Trienyl, 3,7,11,15-tetramethylhexadec-2-enyl and the like, dodecyl, tetradecyl, hexadecyl, (Z) -hexadec-9-enyl, (Z) -octadec-6- More preferred are enyl, (Z) -octadeca-9-enyl, (9Z, 12Z) -octadeca-9,12-dienyl, and the like.

At least one of L ⁸ to L ¹⁰ is -CO-O- (CY ³⁹ Y ⁴⁰ ) _p18- , -CO-NY ^45B- (CY ³⁹ Y ⁴⁰ ) _p18 -or -CO- (CY ³⁹ Y ⁴⁰ ) _p18 In some cases, -CO-O- (CY ³⁹ Y ⁴⁰ ) _p18- , -CO-NY ^45B- (CY ³⁹ Y ⁴⁰ ) _p18 -or -CO- (CY ³⁹ Y ⁴⁰ ) R ⁷ binding to _p18 R ⁹ is the same or different and each represents nonyl, undecyl, tridecyl, pentadecyl, heptadecyl, nonadecyl, henicosyl, tricosyl, (Z) -tridec-8-enyl, (Z) -pentadec-8-enyl, (Z)- Heptadec-5-enyl, (Z) -heptadeca-8-enyl, (E) -heptadeca-8-enyl, (Z) -heptadeca-10-enyl, (8Z, 11Z) -heptadeca-8,11-dienyl, (8Z, 11Z, 14Z) -octadeca-8,11,14-trienyl, (Z) -nonadec-10-enyl, (10Z, 13Z) -nonadec-10,13-dienyl, (11Z, 14Z) -icosa- 11,14-dienyl, 2,6,10-trimethylundeca-1,5,9-trienyl, 2,6,10,14-tetramethylpentadec-1-enyl Preferably, undecyl, tridecyl, pentadecyl, (Z) -pentadec-8-enyl, (Z) -heptadeca-5-enyl, (Z) -heptadeca-8-enyl, (8Z, 11Z) -heptadeca-8, More preferred is 11-dienyl or the like.

L ¹¹ is ^{^{absent, - (CY 46 Y 47)}} p21 -, - (CY 48 Y 49) p22 -Z 15 - (CY 50 Y 51) p23 - or ^{^{_{- (CY 52 Y 53) p24}}} -Z 16 ^{^{_{- (CY 54 Y 55) p25}}} -Z 17 - (CY 56 Y 57) p26 - a is ^{^{absent, - (CY 46 Y 47)}} p21 - or ^{^{_{- (CY 48 Y 49) p22}}} -Z 15 - ( CY ⁵⁰ Y ⁵¹ ) _p23- is preferred. Y ⁴⁶ to Y ⁵⁷ are the same or different and each represents a hydrogen atom or an optionally substituted C1-C4 alkyl, and Y ⁴⁶ to Y ⁵⁷ are preferably a hydrogen atom. Z ¹⁵ ~ Z ¹⁷ are the same or ^{different, -O -, - NY 58A -} , - CO-O -, - O-CO -, - CO-NY 58B -, - NY 58C -CO -, - NY 58D - CO-0- or a -CO-, -CO-O -, - O-CO -, - CO-NY 58B -, - NY 58C -CO- or -CO- are preferable, -O-CO- or - NY ^58C -CO- is more preferred. p ²¹ is an integer of 1 to 5, preferably 1 to 3. p ²² is an integer of 0 to 5, preferably 0 to 3. p ²³ is an integer of 1 to 5, and is preferably 1 or 2.

Do not present as ^{^{L 11, - (CY 46 Y}} 47) p21 -, - (CY 48 Y 49) p22 -O-CO- (CY 50 Y 51) p23 -, or ^{^{_{- (CY 48 Y 49) p22}}} - NY ^58C -CO- (CY ⁵⁰ Y ⁵¹ ) _p23- is preferably absent, more preferably-(CY ⁴⁶ Y ⁴⁷ ) _p21- , absent _or- (CH ₂ ) _p2- More preferably.

L ¹² is preferably absent or — (CY ⁵⁹ Y ⁶⁰ ) _p27 —, preferably absent or — (CH ₂ ) _p27 —, absent, —CH ₂ — or — More preferred is (CH ₂ ) ₂ —.

J ¹ and J ² are the same or different and are CY ⁷² or N, and J ¹ and J ² are the same or different and are preferably CH, C (OH) or N.

When L ¹¹ is not present, J ¹ is preferably CH.

L ⁹ and L ¹⁰ are not present, L ¹² is —CO— (CH ₂ ) _p29 —, J ¹ is CH, and J ² is N, which is one of the preferred embodiments of the present invention. At this time, L ⁸ is preferably —CO—NY ^45B — (CH ₂ ) _p18 —, and L ¹¹ is preferably absent or — (CH ₂ ) _p21 —.

L ⁹ and L ¹⁰ are not present, L ¹² is —O—CO— (CH ₂ ) _p29 —, and J ¹ and J ² are CH. At this time, it is preferable that L ⁸ is —O—CO— (CH ₂ ) _p18 — and L ¹¹ does not exist.

B ²

And is more preferably —N ⁺ (CH ₃ ) ₃ .

X ⁸ to X ¹⁰ have the same meanings as X ² to X ⁴ , respectively.

In the formula (IV), R ¹⁰ is preferably linear or branched C8-C24 alkyl or C8-C24 alkenyl, linear or branched C15-C20 alkenyl, or linear It is more preferably a linear or branched C9-C18 alkyl, most preferably a linear C15-C20 alkenyl, or most preferably a linear C9-C18 alkyl. R ¹¹ and R ¹² are linear or branched C8-C24 alkyl, C8-C24 alkenyl or C8-C24 alkynyl, and are preferably the same, linear or branched C15-C20 alkenyl. Or a linear or branched C9-C18 alkyl and more preferably the same, a linear C15-C20 alkenyl, or a linear C9-C18 alkyl. Most preferably, they are identical.

L ¹³ is ^{^{absent, -Z 23 - (CY 84 Y}} 85) p38 - or ^{^{^{-Z 24 - (CY 86 Y 87}}} ) p39 -Z 25 - (CY 88 Y 89) p40 - a is absent , or ^{^{^{-Z 23 - (CY 84 Y 85}}} ) p38 - is preferably. Y ⁸⁴ to Y ⁸⁹ are the same or different and each represents a hydrogen atom or optionally substituted C1-C4 alkyl, and Y ⁸⁴ to Y ⁸⁹ are preferably hydrogen atoms. Z ²³ ~ Z ²⁵ are the same or ^{different, -O -, - NY 90A -} , - CO-O -, - O-CO -, - CO-NY 90B -, - NY 90C -CO- or -NY ^90D - a CO-0-, -CO-O - , - O-CO -, - CO-NY 90B - or is preferably ^{^{-NY 90C -CO-, -CO-NY 90B}} - and more preferably . Y ^90A to Y ^90D are the same or different and each represents a hydrogen atom or an optionally substituted C1-C4 alkyl, preferably a hydrogen atom or methyl. p ³⁸ to p ⁴⁰ are the same or different and are integers of 1 to 5, preferably 1 or 2.

L ¹³ does not exist or -CO-O- (CY ⁸⁴ Y ⁸⁵ ) _p38- , _-O -CO- (CY ⁸⁴ Y ⁸⁵ ) _p38- , -CO-NY ^90B- (CY ⁸⁴ Y ⁸⁵ ) _p38 -, Or -NY ^90C -CO- (CY ⁸⁴ Y ⁸⁵ ) _p38- , preferably absent or -CO-O- (CH ₂ ) _p38- , -O-CO- (CH ₂ ) _p38- _{Alternatively} , it is more preferably —CO—NCH ₃ — (CH ₂ ) _p38 —, and it is more preferably absent or —CO—NCH ₃ — (CH ₂ ) _p38 —.

L ¹⁴ and L ¹⁵ is absent same or ^{^{different, -Z 26 - (CY 91 Y}} 92) p41 - or ^{^{^{-Z 27 - (CY 93 Y 94}}} ) p42 -Z 28 - (CY 95 Y 96) p43 -, and absent or ^{^{^{-Z 26 - (CY 91 Y 92}}} ) p41 - is preferably. Y ⁹¹ to Y ⁹⁶ are the same or different and each represents a hydrogen atom or an optionally substituted C1-C4 alkyl, and Y ⁹¹ to Y ⁹⁶ are preferably a hydrogen atom. Z ²⁶ ~ Z ²⁸ are the same or ^{different, -O -, - NY 97A -} , - CO-O -, - O-CO -, - CO-NY 97B -, - NY 97C -CO -, - NY 97D - CO-0- or -CO-, preferably -CO-O-, -O-CO-, -CO-NY ^97B- , ^{-NY 97C} -CO-, or -CO-. Y ^97A to Y ^97D are the same or different and are a hydrogen atom or an optionally substituted C1-C4 alkyl, preferably a hydrogen atom or methyl. p ⁴¹ is an integer of 0 to 5, preferably 0 to 2. p ⁴² is an integer of 1 to 5, preferably 1 or 2. p ⁴³ is an integer of 0 to 5, preferably 0 to 2.

L ¹⁴ and L ¹⁵ are the same or different and do not exist or are -CO-O- (CY ⁹¹ Y ⁹² ) _p41- , _-O -CO- (CY ⁹¹ Y ⁹² ) _p41- , -CO-NY ^97B -(CY ⁹¹ Y ⁹² ) _p41- , ^{-NY 97C} -CO- (CY ⁹¹ Y ⁹² ) _p41- , -CO- (CY ⁹¹ Y ⁹² ) _p41- , preferably the same or different and not present , -CO-O- (CY ⁹¹ Y ⁹² ) _p41- , _-O -CO- (CY ⁹¹ Y ⁹² ) _p41 -or -CO- (CY ⁹¹ Y ⁹² ) _p41- , more preferably the same or different More preferably, it is absent, or —CO—O— (CH ₂ ) _p41 —, —O—CO— (CH ₂ ) _p41 — or —CO—.

In the formula (IV), L ¹³ is -CO-O- (CY ⁸⁴ Y ⁸⁵ ) _p38- , _-O -CO- (CY ⁸⁴ Y ⁸⁵ ) _p38 -or -CO-NY ^90B- (CY ⁸⁴ Y ⁸⁵ ) _p38 -Or one of L ¹⁴ and L ¹⁵ is -CO-O- (CY ⁹¹ Y ⁹² ) _p41 -or -O-CO- (CY ⁹¹ Y ⁹² ) _p41- , or L ¹³ is -CO -O- (CY ⁸⁴ Y ⁸⁵ ) _p38- , -O-CO- (CY ⁸⁴ Y ⁸⁵ ) _p38 -or -CO-NY ^89B- (CY ⁸⁴ Y ⁸⁵ ) _p38- , of L ¹⁴ and L ¹⁵ One is -CO-O- (CY ⁹¹ Y ⁹² ) _p41 -or -O-CO- (CY ⁹¹ Y ⁹² ) _p41- , or L ¹⁴ and L ¹⁵ are the same or different and -CO-O- (CY ⁹¹ Y ⁹² ) _p41 -or -O-CO- (CY ⁹¹ Y ⁹² ) _p41- or L ¹³ is -CO-O- (CY ⁸⁴ Y ⁸⁵ ) _p38- , -O-CO- (CY ⁸⁴ Y ⁸⁵ ) _p38 -or -CO-NY ^89B- (CY ⁸⁴ Y ⁸⁵ ) _p38- , and L ¹⁴ and L ¹⁵ are the same or different -CO-O- (CY ⁹¹ Y ⁹² ) _{p41 -or} -O-CO More preferably,-(CY ⁹¹ Y ⁹² ) _p41- and R ¹⁰ to R ¹² are linear or branched C15-C20 alkenyl or C9-C18 alkyl. R ¹¹ and R ¹² are preferably the same.

L ¹³ is ^{^{absent, -O- (CY 84 Y 85)}} p38 -, - NY 90A - (CY 84 Y 85) p38 -, - O-CO- (CY 84 Y 85) p38 -, - NY 90C ^{^{_{-CO- (CY 84 Y 85) p38}}} - or ^{^{-NY 90D -CO-0- (CY 84}} Y 85) p38 - if it is in, R ¹⁰ is octyl, decyl, dodecyl, tetradecyl, hexadecyl, octadecyl, eicosyl , Docosyl, tetracosyl, (Z) -tetradec-9-enyl, (Z) -hexadeca-9-enyl, (Z) -octadeca-6-enyl, (Z) -octadeca-9-enyl, (E) -octadeca -9-enyl, (Z) -octadeca-11-enyl, (9Z, 12Z) -octadeca-9,12-dienyl, (9Z, 12Z, 15Z) -octadeca-9,12,15-trienyl, (Z) -Icosa-11-enyl, (11Z, 14Z) -icosa-11,14-dienyl, 3,7,11-trimethyldodeca-2,6,10-trienyl or 3,7,11,15-tetramethylhexadeca -2-enyl is preferred, dodecyl, tetradecyl, hexadecyl, (Z) -hexadec-9-enyl, (Z) -o Tateshina 6-enyl, (Z) - octadec-9-enyl or (9Z, 12Z) - and more preferably octadeca-9,12-dienyl.

When L ¹³ is -CO-O- (CY ⁸⁴ Y ⁸⁵ ) _p38 -or -CO-NY ^90B- (CY ⁸⁴ Y ⁸⁵ ) _p38- , R ¹⁰ is nonyl, undecyl, tridecyl, pentadecyl, heptadecyl , Nonadecyl, henicosyl, tricosyl, (Z) -tridec-8-enyl, (Z) -pentadeca-8-enyl, (Z) -heptadeca-5-enyl, (Z) -heptadeca-8-enyl, (E) -Heptadeca-8-enyl, (Z) -heptadeca-10-enyl, (8Z, 11Z) -heptadeca-8,11-dienyl, (8Z, 11Z, 14Z) -octadeca-8,11,14-trienyl, ( Z) -nonadeca-10-enyl, (10Z, 13Z) -nonadeca-10,13-dienyl, (11Z, 14Z) -icosa-11,14-dienyl, 2,6,10-trimethylundeca-1,5 , 9-trienyl or 2,6,10,14-tetramethylpentadec-1-enyl, undecyl, tridecyl, pentadecyl, (Z) -pentadec-8-enyl, (Z) -heptadeca- 5-enyl, (Z) -heptadeca-8-enyl or (8Z, 11Z More preferred is) -heptadeca-8,11-dienyl.

Further, at least one of L ¹⁴ and L ^15, ^{^{absent, -O- (CY 91 Y 92)}} p41 -, - NY 97A - (CY 91 Y 92) p41 -, - O-CO- (CY 91 Y ⁹² ) _p41- , -NY ^97C -CO- (CY ⁹¹ Y ⁹² ) _p41 -or -NY ^97D -CO-0- (CY ⁹¹ Y ⁹² ) _p41- , J ³ , -O- (CY ^{^{_{91 Y 92) p41 -, -}}} NY 97A - (CY 91 Y 92) p41 -, - O-CO- (CY 91 Y 92) p41 -, - NY 97C -CO- (CY 91 Y 92) p41 - or - ^{^{NY 97D -CO-0- (CY 91}} Y 92) p41 - R 11 and R ¹² bound to the same or different, octyl respectively, decyl, dodecyl, tetradecyl, hexadecyl, octadecyl, eicosyl, docosyl, tetracosyl, ( (Z) -tetradec-9-enyl, (Z) -hexadeca-9-enyl, (Z) -octadeca-6-enyl, (Z) -octadeca-9-enyl, (E) -octadeca-9-enyl, ( Z) -octadeca-11-enyl, (9Z, 12Z) -octadeca-9,12-dienyl, (9Z, 12Z, 15Z) -octadeca-9,12,15-trienyl, (Z) -icosa-11-enyl , (11Z, 14Z)- More preferred is icosa-11,14-dienyl, 3,7,11-trimethyldodeca-2,6,10-trienyl or 3,7,11,15-tetramethylhexadec-2-enyl, dodecyl, Tetradecyl, hexadecyl, (Z) -hexadec-9-enyl, (Z) -octadeca-6-enyl, (Z) -octadeca-9-enyl or (9Z, 12Z) -octadec-9,12-dienyl Is more preferable.

If at least one of L ¹⁴ and L ¹⁵ is -CO-O- (CY ⁹¹ Y ⁹² ) _p41 -or -CO-NY ^97B (CY ⁹¹ Y ⁹² ) _p41- , -CO-O- (CY ⁹¹ Y ⁹²⁾ _p41 - or ^{^{-CO-NY 97B (CY 91 Y}} 92) p41 - R 11 and R ¹² bind to the same or different and nonyl respectively, undecyl, tridecyl, pentadecyl, heptadecyl, nonadecyl, henicosyl, Tricosyl, (Z) -Tridec-8-enyl, (Z) -pentadeca-8-enyl, (Z) -heptadeca-5-enyl, (Z) -heptadeca-8-enyl, (E) -heptadeca-8- Enyl, (Z) -heptadeca-10-enyl, (8Z, 11Z) -heptadeca-8,11-dienyl, (8Z, 11Z, 14Z) -octadeca-8,11,14-trienyl, (Z) -nonadeca- 10-enyl, (10Z, 13Z) -nonadeca-10,13-dienyl, (11Z, 14Z) -icosa-11,14-dienyl, 2,6,10-trimethylundeca-1,5,9-trienyl or More preferably 2,6,10,14-tetramethylpentadec-1-enyl, Ndecyl, tridecyl, pentadecyl, (Z) -pentadec-8-enyl, (Z) -heptadec-5-enyl, (Z) -heptadec-8-enyl or (8Z, 11Z) -heptadec-8,11-dienyl More preferably it is.

Or L ¹⁶ is ^{^{absent, - (CY 98 Y 99)}} p44 -, - (CY 100 Y 101) p45 -Z 29 - (CY 102 Y 103) p46 - or ^{^{_{- (CY 104 Y 105) p47}}} -Z 30 - ^{^{_{(CY 106 Y 107) p48 -Z}}} 31 - (CY 108 Y 109) p49 - a is ^{^{absent, - (CY 98 Y 99)}} p44 - or ^{^{_{- (CY 100 Y 101) p45}}} -Z 29 - (CY ¹⁰² Y ¹⁰³ ) _p46- , preferably absent,-(CY ⁹⁸ Y ⁹⁹ ) _p44 -,-(CY ¹⁰⁰ Y ¹⁰¹ ) _p45 -O-CO- (CY ¹⁰² Y ¹⁰³ ) _p46 -,-( CY ¹⁰⁰ Y ¹⁰¹ ) _p45 -NY ^109C -CO- (CY ¹⁰² Y ¹⁰³ ) _p46 -or -CO- (CY ¹⁰² Y ¹⁰³ ) _p46- is more preferable and absent _or- (CH ₂ ) _p44- or -CO- (CH ₂₎ _p46 - it is more preferable.

J ³ is CY ¹¹¹ or N, preferably CH or N. Also, when J ³ is N, L ¹⁴ is not present, L ¹⁵ is -CO-, L ¹⁶ is not present, or-(CY ⁹⁸ Y ⁹⁹ ) _p44- or L ¹⁴ More preferably, L ¹⁵ is not present, and L ¹⁶ is —CO— (CY ¹⁰² Y ¹⁰³ ) _p46 —.

X ¹⁴ and X ¹⁵ have the same meanings as X ² and X ³ , respectively.

In the formula (V ′), R ¹³ is preferably linear or branched C8-C24 alkyl or C8-C24 alkenyl, linear or branched C15-C20 alkenyl, or linear It is more preferably a linear or branched C9-C18 alkyl, most preferably a linear C15-C20 alkenyl, or most preferably a linear C9-C18 alkyl. R ¹⁴ and R ¹⁵ are linear or branched C8-C24 alkyl, C8-C24 alkenyl or C8-C24 alkynyl, and are preferably the same, linear or branched C15-C20 alkenyl. Or a linear or branched C9-C18 alkyl and more preferably the same, a linear C15-C20 alkenyl, or a linear C9-C18 alkyl. Most preferably, they are identical.

L ¹⁷ ~ L ¹⁹ is absent same or ^{^{different, -Z 32 - (CY 116 Y}} 117) p51 - or ^{^{^{-Z 33 - (CY 118 Y 119}}} ) p52 -Z 34 - (CY 120 Y 121) p53 -Z, -Z ^32- (CY ¹¹⁶ Y ¹¹⁷ ) _p51- , preferably -O- (CY ¹¹⁶ Y ¹¹⁷ ) _p51 -or -CO-O- (CY ¹¹⁶ Y ¹¹⁷ ) _p51- Is more preferable, and —O— or —CO—O— is more preferable.

In Formula (V ′), L ¹⁷ to L ¹⁹ are the same or different and are —O— or —CO—O—, and R ¹³ to R ¹⁵ are linear C15-C20 alkenyl or C9-C18 alkyl. Is preferred. At this time, it is preferable that L ¹⁷ to L ¹⁹ are identically —O— or —CO—O—, and R ¹³ to R ¹⁵ are identically linear C15-C20 alkenyl or C9-C18 alkyl.

At least one of L ¹⁷ to L ¹⁹ does not exist or is -O- (CY ¹¹⁶ Y ¹¹⁷ ) _p51- , -O-CO- (CY ¹¹⁶ Y ¹¹⁷ ) _p51- , ^{-NY 122C} -CO- (CY ¹¹⁶ Y ¹¹⁷⁾ _P518 - or ^{^{-NY 122D -CO-0- (CY 116}} Y 117) p51 - a if the carbon adjacent to the furanose ring or ^{^{L 20, -O- (CY 116 Y}} 117) p51 -, - ^{^{O-CO- (CY 116 Y 117}} ) p51 -, - NY 122C -CO- (CY 116 Y 117) p518 - or ^{^{-NY 122D -CO-0- (CY 116}} Y 117) p51 - binding to R ¹³ ~ R ¹⁵ is the same or different and is octyl, decyl, dodecyl, tetradecyl, hexadecyl, octadecyl, icosyl, docosyl, tetracosyl, (Z) -tetradec-9-enyl, (Z) -hexadec-9-enyl, (Z ) -Octadeca-6-enyl, (Z) -octadeca-9-enyl, (E) -octadeca-9-enyl, (Z) -octadeca-11-enyl, (9Z, 12Z) -octadeca-9,12- Dienyl, (9Z, 12Z, 15Z) -octadeca-9,12,15-trienyl, (Z) -icosa-11-enyl, (11Z, 14Z) -ico -11,14-dienyl, 3,7,11-trimethyldodeca-2,6,10-trienyl, 3,7,11,15-tetramethylhexadec-2-enyl, etc., preferably dodecyl, tetradecyl , Hexadecyl, (Z) -hexadec-9-enyl, (Z) -octadeca-6-enyl, (Z) -octadeca-9-enyl, (9Z, 12Z) -octadec-9,12-dienyl, etc. Is more preferable.

At least one of L ¹⁷ to L ¹⁹ is -CO-O- (CY ¹¹⁶ Y ¹¹⁷ ) _p51- , -CO-NY ^122B- (CY ¹¹⁶ Y ¹¹⁷ ) _p51 -or -CO- (CY ¹¹⁶ Y ¹¹⁷ ) _p51- R is bound to -CO-O- (CY ¹¹⁶ Y ¹¹⁷ ) _p51- , -CO-NY ^122B- (CY ¹¹⁶ Y ¹¹⁷ ) _p51 -or -CO- (CY ¹¹⁶ Y ¹¹⁷ ) _p51- ¹³ to R ¹⁵ are the same or different and are each nonyl, undecyl, tridecyl, pentadecyl, heptadecyl, nonadecyl, henicosyl, tricosyl, (Z) -tridec-8-enyl, (Z) -pentadec-8-enyl, (Z ) -Heptadeca-5-enyl, (Z) -heptadeca-8-enyl, (E) -heptadeca-8-enyl, (Z) -heptadeca-10-enyl, (8Z, 11Z) -heptadeca-8,11- Dienyl, (8Z, 11Z, 14Z) -octadeca-8,11,14-trienyl, (Z) -nonadec-10-enyl, (10Z, 13Z) -nonadec-10,13-dienyl, (11Z, 14Z)- Icosa-11,14-dienyl, 2,6,10-trimethylundeca-1,5,9-trienyl, 2,6,10,14-tetramethylpentadeca 1-enyl is preferred, undecyl, tridecyl, pentadecyl, (Z) -pentadec-8-enyl, (Z) -heptadeca-5-enyl, (Z) -heptadeca-8-enyl, (8Z, 11Z More preferred is) -heptadeca-8,11-dienyl.

L ²⁰ is ^{^{absent, - (CY 123 Y 124)}} p54 -, - (CY 125 Y 126) p55 -Z 35 - (CY 127 Y 128) p56 - or ^{^{_{- (CY 129 Y 130) p57}}} -Z 36 ^{^{_{- (CY 131 Y 132) p58}}} -Z 37 - (CY 133 Y 134) p59 - a and, - is ^{^{preferably, - - (CY 123 Y 124}} ) p54 (CH 2) p54 - a is more Preferably, it is —CH ₂ —.

L ²¹ is ^{^{absent, - (CY 136 Y 137)}} p60 -, - (CY 138 Y 139) p61 -Z 38 - (CY 140 Y 141) p62 - or ^{^{_{- (CY 142 Y 143) p63}}} -Z 39 -(CY ¹⁴⁴ Y ¹⁴⁵ ) _p64 -Z ^40- (CY ¹⁴⁶ Y ¹⁴⁷ ) _p65- , preferably absent _or- (CY ¹³⁶ Y ¹³⁷ ) _p60- , absent _or- (CH ₂ ) More preferred is _p60 -and even more preferred is absent.

B ³

And is more preferably —N ⁺ (CH ₃ ) ₃ .

Y ¹¹² and Y ¹¹³ are the same or different and are a hydrogen atom, hydroxy or optionally substituted C1-C4 alkyl, the same or different, preferably a hydrogen atom or hydroxy, and more preferably a hydrogen atom.

In the formula (V ''), R ¹⁶ to R ¹⁸ , L ²² to L ²⁶ , B ⁴ , Y ¹¹⁴ to Y ¹¹⁵ and A ⁶ are R ¹³ to R ¹⁵ , L ¹⁷ to L ²¹ , B ³ and Y, respectively. ¹¹² to Y ¹¹³ and A ⁵ are synonymous.

In formula (V ′), when Y ¹¹² is a hydrogen atom, the four substituents on the pyran ring are preferably substituted on different carbon atoms on the pyran ring. And as a formula (V '),

It is more preferable that At this time, L ¹⁷ to L ¹⁹ are the same or different and are —O— or —CO—O—, and R ¹³ to R ¹⁵ are more preferably linear C15-C20 alkenyl or C9-C18 alkyl, L ¹⁷ to L ¹⁹ are the same or different and are —O— or —CO—O—, R ¹³ to R ¹⁵ are linear C15-C20 alkenyl or C9-C18 alkyl, and L ¹⁷ and L ²¹ are present Most preferably, Y ¹¹³ is a hydrogen atom or hydroxy.

In the formula (V ″), it is preferable that each of the four substituents on the furan ring is substituted on a different carbon atom on the furan ring. And as formula (V ''),

It is more preferable that At this time, it is more preferable that L ²² to L ²⁴ are the same or different and are —O— or —CO—O—, and R ¹⁶ to R ¹⁸ are linear C15-C20 alkenyl or C9-C18 alkyl, L ²² to L ²⁴ are the same or different and are —O— or —CO—O—, R ¹⁶ to R ¹⁸ are linear C15-C20 alkenyl or C9-C18 alkyl, and L ²² and L ²⁶ are present. Most preferably, Y ¹¹⁴ is a hydrogen atom or hydroxy.

In the definitions of the formulas B ¹ , B ² , B ³ and B ⁴ ,

In the definition of each formula, when p ¹³ , p ³³ and p ⁶⁶ are 0, N ⁺ is bonded to carbon adjacent to Z ¹⁰ , Z ²¹ and Z ⁴¹ , respectively.

The nucleic acid-containing lipid nanoparticles of the present invention include a cationic lipid other than a lipid having a hydrophilic part having one quaternary ammonium group and three independent hydrocarbon groups which may be substituted (lipid A). May be. Cationic lipids other than lipid A used in the present invention include a lipophilic region containing one or more optionally substituted hydrocarbon groups, at least one primary amino group, secondary amino group, and tertiary amino group. Any amphiphilic molecule having a cationic hydrophilic region containing a group and / or a quaternary ammonium group (except for lipid A), one amino group or one 4 which may be substituted A lipid (lipid B) containing a hydrophilic part having a quaternary ammonium group and a hydrophobic part having two optionally substituted independent hydrocarbon groups in the same molecule is preferred.

In the present invention, a lipid having a hydrophilic part having one amino group or one quaternary ammonium group which may be substituted and a hydrophobic part having two independent hydrocarbon groups which may be substituted in the same molecule (Lipid B) has one amino group or one quaternary ammonium group that may be substituted as a hydrophilic part in the molecule, and two independent hydrocarbon groups that may be substituted. Although it will not be restrict | limited especially if it is a molecule | numerator, For example, it represents with following Structural formula (D) and (E).
In the following structural formulas (D) and (E), “hydrophilic unit” represents a hydrophilic part having one amino group or one quaternary ammonium group which may be substituted, and “hydrophobic part ( Hydrophobic Unit) "represents an independent hydrocarbon group which may be substituted.

The amino group constituting the “Hydrophilic Unit” has 0 to 2 of the three bonds, and 0 to 2 hydrocarbon groups forming the “Hydrophobic Unit”. And the remaining bonds are bonded to hydrogen or an optionally substituted chain and / or cyclic hydrocarbon group.
In addition, the quaternary ammonium group constituting the “hydrophilic unit” is one of hydrocarbon groups in which 0 to 2 of the four bonds form a “hydrophobic unit”. 0 to 2 bonds are bonded, and the remaining bond is bonded to hydrogen or an optionally substituted chain and / or cyclic hydrocarbon group.
The optionally substituted chain and / or cyclic hydrocarbon group constituting the “hydrophilic unit” may be any group consisting of a carbon atom and a hydrogen atom. Those having 10 carbon atoms are preferable, those having 1 to 6 carbon atoms are more preferable, and those having 1 to 3 carbon atoms are more preferable.

The hydrocarbon group forming the “Hydrophobic Unit” may be directly bonded to the amino group or quaternary ammonium group of the “Hydrophilic Unit”, or may be a bond such as ether, ester or amide. And may be bonded to an amino group or a quaternary ammonium group via a chain and / or a cyclic hydrocarbon group which may have a substituent constituting the “hydrophilic part”. Further, as shown in the structural formula (E), hydrocarbon groups forming two “hydrophobic units” are bonded via a carbon atom, and the carbon atom is “hydrophilic unit”. ) "Amino group or quaternary ammonium group, or a chain and / or cyclic hydrocarbon group which may have a substituent such as a bond of ether, ester, amide or the like and a" hydrophilic part " And may be bonded to an amino group or a quaternary ammonium group.

Examples of cationic lipids other than lipid A used in the present invention include, for example, International Publication No. 2013/089151, International Publication No. 2011/136368, International Publication No. 2014/007398, International Publication No. 2010/042877 or International Publication No. And cationic lipids described in JP 2010/054401 A.

As lipid B used in the present invention, for example,
Formula (CL-I)

(Where
R ¹⁰¹ and R ¹⁰² are the same or different and are linear or branched C10-C24 alkyl, C10-C24 alkenyl or C10-C24 alkynyl,
L ¹⁰¹ and L ¹⁰² are hydrogen atoms or together form a single bond or C1-C3 alkylene;
L ¹⁰³ is a single bond, -CO- or -CO-O-,
When L ¹⁰³ is a single bond,
X ¹⁰¹ is a hydrogen atom, C1-C6 alkyl, C3-C6 alkenyl, pyrrolidin-3-yl, piperidin-3-yl, piperidin-4-yl, or the same or different 1 to 3 amino, monoalkylamino, dialkyl C1-C6 alkyl or C3-C6 alkenyl substituted with amino, trialkylammonio, hydroxy, alkoxy, carbamoyl, monoalkylcarbamoyl, dialkylcarbamoyl, pyrrolidinyl, piperidyl or morpholinyl,
When L ¹⁰³ is -CO- or -CO-O-
X ¹⁰¹ is pyrrolidin-2-yl, pyrrolidin-3-yl, piperidin-2-yl, piperidin-3-yl, piperidin-4-yl, morpholin-2-yl, morpholin-3-yl, or the same or different With 1 to 3 amino, monoalkylamino, dialkylamino, trialkylammonio, hydroxy, alkoxy, carbamoyl, monoalkylcarbamoyl, dialkylcarbamoyl, pyrrolidinyl, piperidyl or morpholinyl substituted C1-C6 alkyl or C3-C6 alkenyl And at least one of the substituents is amino, monoalkylamino, dialkylamino, trialkylammonio, pyrrolidinyl, piperidyl or morpholinyl),
Formula (CL-II)

(Where
R ¹⁰³ and R ¹⁰⁴ are the same or different and are linear or branched C12-C24 alkyl, C12-C24 alkenyl or C12-C24 alkynyl,
p ¹⁰¹ and p ¹⁰² are the same or different and are integers of 0 to 3,
L ¹⁰⁶ and L ¹⁰⁷ are hydrogen atoms or together form a single bond or C2-C8 alkylene;
L ¹⁰⁴ and L ¹⁰⁵ are the same or different and are -O-, -CO-O- or -O-CO-,
L ¹⁰⁸ is a single bond, -CO- or -CO-O-
When L ¹⁰⁸ is a single bond,
X ¹⁰² is a hydrogen atom, C1-C6 alkyl, C3-C6 alkenyl, pyrrolidin-2-yl, pyrrolidin-3-yl, piperidin-3-yl, piperidin-4-yl, or the same or different 1 to 3 amino acids C1-C6 alkyl or C3-C6 alkenyl substituted with monoalkylamino, dialkylamino, trialkylammonio, hydroxy, alkoxy, carbamoyl, monoalkylcarbamoyl, dialkylcarbamoyl, pyrrolidinyl, piperidyl or morpholinyl,
When L ¹⁰⁸ is -CO- or -CO-O-
X ¹⁰² is pyrrolidin-2-yl, pyrrolidin-3-yl, piperidin-2-yl, piperidin-3-yl, piperidin-4-yl, morpholin-2-yl, morpholin-3-yl, or the same or different With 1 to 3 amino, monoalkylamino, dialkylamino, trialkylammonio, hydroxy, alkoxy, carbamoyl, monoalkylcarbamoyl, dialkylcarbamoyl, pyrrolidinyl, piperidyl or morpholinyl substituted C1-C6 alkyl or C3-C6 alkenyl And at least one of the substituents is amino, monoalkylamino, dialkylamino, trialkylammonio, pyrrolidinyl, piperidyl or morpholinyl),
Formula (CL-III)

(Where
R ¹⁰⁵ is linear or branched C8-C24 alkyl, C8-C24 alkenyl or C8-C24 alkynyl,
R ¹⁰⁶ is linear or branched C8-C24 alkyl, C8-C24 alkenyl, C8-C24 alkynyl, C8-C24 alkyloxyethyl, C8-C24 alkyloxypropyl, C8-C24 alkenyloxyethyl, C8-C24 alkenyl Oxypropyl, C8-C24 alkynyloxyethyl or C8-C24 alkynyloxypropyl,
X ¹⁰³ and X ¹⁰⁴ are the same or different and are C1-C3 alkyl or taken together to form C2-C8 alkylene, or X ¹⁰³ is taken together with L ¹¹¹ to form C2-C8 alkylene. ,
L ¹¹¹ is a hydrogen atom, C1-C6 alkyl, C3-C6 alkenyl, amino, monoalkylamino, hydroxy, alkoxy, carbamoyl, monoalkylcarbamoyl, dialkylcarbamoyl, or the same or different 1 to 3 amino, monoalkylamino, C1-C6 alkyl or C3-C6 alkenyl substituted with hydroxy, alkoxy, carbamoyl, monoalkylcarbamoyl or dialkylcarbamoyl, or together with X ¹⁰³ to form C2-C8 alkylene;
L ¹⁰⁹ is C1-C6 alkylene,
L ¹¹⁰ is a single bond or C1-C6 alkylene, provided that the sum of the carbon numbers of L ¹⁰⁹ and L ¹¹⁰ is 7 or less, and when L ¹¹¹ is a hydrogen atom, L ¹¹⁰ is a single bond. And when L ¹¹¹ together with X ¹⁰³ forms C2-C6 alkylene, L ¹¹⁰ is a single bond or is methylene or ethylene),
Formula (CL-IV)

(Where
R ¹⁰⁷ is linear or branched C8-C24 alkyl, C8-C24 alkenyl or C8-C24 alkynyl,
R ¹⁰⁸ is linear or branched C8-C24 alkyl, C8-C24 alkenyl, C8-C24 alkynyl, C8-C24 alkyloxyethyl, C8-C24 alkyloxypropyl, C8-C24 alkenyloxyethyl, C8-C24 alkenyl Oxypropyl, C8-C24 alkynyloxyethyl, C8-C24 alkynyloxypropyl, C8-C24 alkyloxyethoxyethyl, C8-C24 alkenyloxyethoxyethyl or C8-C24 alkynyloxyethoxyethyl,
X ¹⁰⁵ is a hydrogen atom, optionally substituted C1-C4 alkyl or -CO- (CH ₂ ) _n -NY1Y2,
n represents an integer of 1 to 4,
Y1 and Y2 are the same or different and are C1-C3 alkyl or together form C2-C8 alkylene)
Formula (CL-V)

(Where
R ¹⁰⁹ is linear or branched C8-C24 alkyl, C8-C24 alkenyl or C8-C24 alkynyl,
R ¹¹⁰ is linear or branched C8-C24 alkyl, C8-C24 alkenyl, C8-C24 alkynyl, C8-C24 alkyloxyethyl, C8-C24 alkyloxypropyl, C8-C24 alkenyloxyethyl, C8-C24 alkenyl Oxypropyl, C8-C24 alkynyloxyethyl or C8-C24 alkynyloxypropyl,
L ¹¹² is C1-C3 alkylene;
X ¹⁰⁵ ′ is a hydrogen atom or C1-C3 alkyl),
Formula (CL-VI)

(Where
R ¹¹¹ and R ¹¹² are the same or different, linear or branched optionally substituted C8-C24 alkyl, C8-C24 alkenyl or C8-C24 alkynyl,
X ¹⁰⁶ and X ¹⁰⁷ are the same or different and are C1-C3 alkyl or taken together to form C2-C8 alkylene;
p ¹⁰³ , p ¹⁰⁴ and p ¹⁰⁵ are the same or different and are 0 or 1, provided that p ¹⁰³ , p ¹⁰⁴ and p ¹⁰⁵ are not 0 at the same time,
L ¹¹³ and L ¹¹⁴ are the same or different and are O, S or NH),
Formula (CL-VII)

(Where
R ¹¹³ and R ¹¹⁴ are the same or different, linear or branched optionally substituted C8-C24 alkyl, C8-C24 alkenyl or C8-C24 alkynyl,
R ¹¹⁵ is a hydrogen atom, hydroxy, optionally substituted C1-C4 alkyl, C1-C4 alkoxy or C1-C4 acyloxy,
X ¹⁰⁹ and X ¹¹⁰ are the same or different and are C1-C3 alkyl or taken together to form C2-C8 alkylene;
L ¹¹⁵ is -CO-O-, -O-CO-, -NHCO- or -CONH-
p ¹⁰⁶ is an integer from 0 to 3,
p ¹⁰⁷ is an integer of 1 to 4),
Formula (CL-VIII)

(Where
R ¹¹⁶ and R ¹¹⁷ are the same or different and may be linear or branched substituted C8-C24 alkyl, C8-C24 alkenyl, C8-C24 alkynyl, C7-C20 alkyloxy C1-C3 alkyl, C7- C20 alkenyloxy C1-C3 alkyl or C7-C20 alkynyloxy C1-C3 alkyl,
Said C8-C24 alkyl, C8-C24 alkenyl or C8-C24 alkynyl having a biodegradable group incorporated therein, or C8-C24 alkyl group, C8-C24 alkenyl group or C8- A C24 alkynyl group,
The biodegradable group is incorporated as —C (O) O— or —OC (O) —, and present at the terminal is —C (O) O—C1-C4 alkyl or -OC (O) -C1-C4 alkyl,
B ¹⁰⁰ is a hydrogen atom, C1-C3 alkyl, hydroxy C2-C4 alkyl, C1-C3 dialkylamino C2-C4 alkyl, formula (A)

(In the formula, X ¹¹¹ and X ¹¹² are the same or different and are a hydrogen atom or C1-C3 alkyl, or together with the nitrogen atom to which X ¹¹¹ and X ¹¹² are bonded, a C2-C6 nitrogen-containing heterocycle is formed. P ¹¹⁰ is an integer from 2 to 6), or formula (B)

(Wherein X ¹¹³ and X ¹¹⁴ are the same or different and are a hydrogen atom or C1-C3 alkyl, or together with the nitrogen atom to which X ¹¹³ and X ¹¹⁴ are bonded form a C2-C6 nitrogen-containing heterocycle And p ¹¹¹ is an integer from 1 to 6),
p ¹⁰⁸ is an integer from 0 to 4, p ¹⁰⁹ is an integer from 1 to 4 (except when p ¹⁰⁸ is 0 and p ¹⁰⁹ is 1),
L ¹¹⁶ is the same or different for each carbon to be bonded and is a hydrogen atom or C1-C3 alkyl;
L ¹¹⁷ is the same or different for each carbon to be bonded and is a hydrogen atom or C1-C3 alkyl).
Formula (CL-IX)

(Where
X ¹¹⁵ and X ¹¹⁶ are the same or different and are a hydrogen atom or C1-C3 alkyl,
L ¹¹⁸ and L ¹¹⁹ are the same or different and are linear or branched C8-C24 alkylene or C8-C24 alkenylene,
M ¹⁰¹ and M ¹⁰² are the same or different, and -C = C-, -OC (O)-, -C (O) O-, -SC (O)-, -C (O) S-, -OC (S )-, -C (S) O-, -SS-, -C (R ^'' ) = N-, -N = C (R ^'' )-, -C (R ^'' ) = NO-, -ON = C (R ^'' )-, -N (R ^'' ) C (O)-, -C (O) N (R ^'' )-, -N (R ^'' ) C (S)-, -C (S) N (R ^'' )-, -N (R ^'' ) C (O) N (R ^''' )-, -N (R ^'' ) C (O) O-, -OC (O) Selected from the group consisting of N (R ^'' )-and -OC (O) O-
R ^'' and R ^''' are the same or different and are a hydrogen atom or C1-C3 alkyl;
R ¹¹⁸ and R ¹¹⁹ are the same or different and are linear or branched optionally substituted C1-C16 alkyl or C2-C16 alkenyl),
Formula (CL-X)

(Where
X ¹¹⁷ and X ¹¹⁸ are the same or different hydrogen atoms, optionally substituted C1-C6 alkyl, heterocyclyl or polyamine, or X ¹¹⁷ and X ¹¹⁸ together with the nitrogen to which they are attached, In addition to nitrogen, it may form a 4-7 membered monocyclic heterocycle which may contain 1 or 2 additional heteroatoms selected from N, O and S;
R ¹²⁰ and R ¹²¹ are the same or different and are linear or branched optionally substituted C4-C24 alkyl or C4-C24 alkenyl),
Formula (CL-XI)

(Where
X ¹¹⁹ and X ¹²⁰ are the same or different and each represents a hydrogen atom, a linear or branched C1-C20 alkyl, C1-C20 alkenyl, C1-C20 alkynyl or C6-C20 acyl,
R ¹²² and R ¹²³ are the same or different, linear or branched optionally substituted C1-C30 alkyl, C2-C30 alkenyl or C2-C30 alkynyl,
p ¹¹² , p ¹¹³ and p ¹¹⁴ are the same or different and are 0 or any positive integer. ),
Formula (CL-XII)

(Where
X ¹²¹ and X ¹²² are the same or different and each is a hydrogen atom, C1-C6 alkyl, cycloalkyl, cycloalkenyl, or X ¹²¹ and X ¹²² together with the nitrogen atom to which they are attached contain C2-C6 nitrogen A heterocycle may form,
L ¹²⁰ and L ¹²¹ are the same or different and are —O—, —OC (O) — or — (O) CO—,
R ¹²⁴ and R ¹²⁵ are the same or different and are linear or branched optionally substituted C8-C24 alkyl or C8-C24 alkenyl),
Formula (CL-XIII)

(Where
R ¹²⁶ and R ¹²⁷ are the same or different and may be linear or branched substituted C8-C24 alkyl, C8-C24 alkenyl, C8-C24 alkynyl, C8-C24 heteroalkyl, C8-C24 heteroalkenyl or C8-C24 heteroalkynyl,
X ¹²³ is a hydrogen atom or an optionally substituted C1-C6 alkyl,
X ¹²⁴ is C1-C6 alkyl, substituted C1-C6 alkyl substituted with -NR ^4a R ^4b or optionally substituted C3-C7 heterocyclyl;
R ^4a and R ^4b are the same or different and are a hydrogen atom, -C (= NH) NH ₂ or an optionally substituted C1-C6 alkyl, or R ^4a and R ^4b are nitrogen atoms to which they are bonded. Together with C3-C7 heterocyclyl, which may be substituted,
X ¹²³ and X ¹²⁴ together with the nitrogen atom to which they are attached may form an optionally substituted C3-C7 heterocyclyl;
However, X ¹²³ and X ¹²⁴ do not form imidazolyl, benzimidazolyl, or succinimidyl, and only one primary amine can be present on either of X ¹²³ and X ¹²⁴ , or any primary amines also not present on either one of X ¹²³ and X ^124, X ¹²³ and X ¹²⁴ is not substituted amides,
When R ¹²⁶ and R ¹²⁷ are C11 alkyl or C15 alkyl, X ¹²³ is not a hydrogen atom,
When R ¹²⁶ and R ¹²⁷ are C16 alkyl or C17 alkyl, R ¹²⁶ and R ¹²⁷ are not substituted with OH;
When R ¹²⁶ and R ¹²⁷ are C17 alkyl, X ¹²³ and X ¹²⁴ are not substituted with OH;
When R ¹²⁶ and R ¹²⁷ are C18 alkyl, X ¹²⁴ is not substituted with an optionally substituted imidazolyl)
Formula (CL-XIV)

(Where
X ¹²⁵ and X ¹²⁶ are the same or different hydrogen atoms, optionally substituted C1-C6 alkyl, heterocyclyl or polyamine, or X ¹²⁵ and X ¹²⁶ together with the nitrogen to which they are attached, In addition to nitrogen, it may form a 4-7 membered monocyclic heterocycle which may contain 1 or 2 additional heteroatoms selected from N, O and S;
R ¹³⁰ is a hydrogen atom or C1-C6 alkyl;
R ¹²⁸ and R ¹²⁹ are the same or different, linear or branched optionally substituted C4-C24 alkyl or C4-C24 alkenyl,
Y3 and Y4 represents an oxygen atom or CH ₂ identical or different,
p115 is an integer of 0-2. ),
Formula (CL-XV)

(Where
X ¹²⁷ and X ¹²⁸ are the same or different and are C1-C6 alkyl, C2-C6 alkenyl, C2-C6 alkynyl,
X ¹²⁷ and X ¹²⁸ together with the nitrogen atom to which they are attached form a heterocyclic ring having 1 to 2 nitrogen atoms;
L ¹²² is -C (O) O-, -OC (O)-, -C (O) N (X ¹³⁰ )-, -N (X ¹³⁰ ) C (O)-, -OC (O) O-, -OC (O) N (X ¹³⁰ )-, -N (X ¹³⁰ ) C (O) N (X ¹³⁰ )-, or -N (X ¹³⁰ ) C (O) O-
Each ^occurrence of X ¹³⁰ is independently a hydrogen atom or C1-C3 alkyl;
a is 1, 2, 3, 4, 5, or 6;
b is 0, 1, 2, or 3;
X ¹²⁹ is absent or is hydrogen or C1-C3 alkyl;
R ¹³¹ and R ¹³² are the same or different and are alkyl having 12 to 24 carbon atoms, alkenyl having 12 to 24 carbon atoms, or alkoxy having 12 to 24 carbon atoms having one or more biodegradable groups, The decomposable group is independently incorporated into the alkyl group having 12 to 24 carbon atoms, the alkenyl group having 12 to 24 carbon atoms, or the alkoxy group having 12 to 24 carbon atoms, or an alkyl group having 12 to 24 carbon atoms. , At the end of an alkenyl group having 12 to 24 carbon atoms or an alkoxy group having 12 to 24 carbon atoms,
The biodegradable group is incorporated as —C (O) O—, —OC (O) —, —C (O) N (X ¹³⁰ ) —, or —N (X ¹³⁰ ) C ( O)-and present at the terminal are -C (O) O-C1-C4 alkyl, -OC (O) -C1-C4 alkyl, -C (O) N (X ¹³⁰ ) -C1-C4 Alkyl, or -N (X ¹³⁰ ) C (O) -C1-C4 alkyl,
R ¹³¹ and R ¹³² have at least 4 carbon atoms between the biodegradable group and a tertiary carbon atom marked with an asterisk (*)),
Formula (CL-XVI)

(Where
R ¹³³ and R ¹³⁴ are the same or different and are linear or branched C1-C9 alkyl, C2-C11 alkenyl or C2-C11 alkynyl;
L ¹²³ and L ¹²⁴ are the same or different and are linear C5-C18 alkylene or linear C5-C18 alkenylene, or form a heterocyclic ring with N bonded thereto,
L ¹²⁵ is a single bond or -C (O) -O-;
When L ¹²⁵ is -C (O) O-, -L ¹²⁴ -CO-OR ¹³⁴ is formed,
L ¹²⁷ is S or O,
L ¹²⁶ is a single bond, a linear or branched C1-C6 alkylene, or forms a heterocyclic ring with N bonded via -C (O)-,
L ¹²⁸ is a linear or branched C1-C6 alkylene, and X ¹³¹ and X ¹³² are the same or different and each is hydrogen or a linear or branched C1-C6 alkyl) or formula ( CL-XVII)

(Where
L ¹³¹ is C2-C4 len or -CH ₂ -S-CH ₂ CH _2-
L ¹²⁹ and L ¹³⁰ are the same or different and each is C1-C6 alkyl;
R ¹³⁵ and R ¹³⁶ are the same or different and are C10-C30 alkyl, C10-C30 alkenyl,
X ¹³³ and X ¹³⁴ are the same or different and are hydrogen, C1-C6 alkyl or —CH ₂ CH ₂ OH. ) And the like.

In the definition of each group of the formula (CL-I), examples of the linear or branched C10-C24 alkyl include decyl, undecyl, dodecyl, tridecyl, 6,10-dimethylundec-2-yl, tetradecyl, Pentadecyl, hexadecyl, heptadecyl, octadecyl, 6,10,14-trimethylpentadecan-2-yl, nonadecyl, icosyl, henicosyl, docosyl, tricosyl, tetracosyl and the like, preferably decyl, undecyl, dodecyl, tridecyl, tetradecyl, Examples include pentadecyl, hexadecyl, heptadecyl, and octadecyl, and more preferable examples include tridecyl, tetradecyl, pentadecyl, hexadecyl, heptadecyl, and octadecyl.

The linear or branched C10-C24 alkenyl may be a linear or branched C10-C24 alkenyl containing 1 to 3 double bonds, such as (Z) -dodec-7-enyl. , (Z) -tetradec-7-enyl, (Z) -tetradec-9-enyl, (Z) -hexadec-4-enyl, (Z) -hexadeca-7-enyl, (E) -hexadeca-7-enyl , (Z) -hexadec-9-enyl, (7Z, 10Z) -hexadec-7,10-dienyl, (7Z, 10Z, 13Z) -hexadec-7,10,13-trienyl, (Z) -octadeca-6 -Enyl, (Z) -octadeca-9-enyl, (E) -octadeca-9-enyl, (9Z, 12Z) -octadeca-9,12-dienyl, (Z) -octadeca-11-enyl, (9Z, 12Z, 15Z) -octadeca-9,12,15-trienyl, (Z) -icosa-11-enyl, (11Z, 14Z) -icosa-11,14-dienyl, etc., preferably (Z) -dodeca -7-enyl, (Z) -tetradec-7-enyl, (Z) -hexadec-4-enyl, (Z) -hexadec-7-enyl, (E) -hexa Deca-7-enyl, (Z) -hexadec-9-enyl, (7Z, 10Z) -hexadec-7,10-dienyl, (7Z, 10Z, 13Z) -hexadec-7,10,13-trienyl, (Z ) -Octadeca-9-enyl, (9Z, 12Z) -octadeca-9,12-dienyl, (11Z, 14Z) -icosa-11,14-dienyl, and more preferably (7Z, 10Z) -hexadeca -7,10-dienyl or (9Z, 12Z) -octadeca-9,12-dienyl.

The linear or branched C10-C24 alkynyl may be a linear or branched C10-C24 alkynyl containing 1 to 3 triple bonds, and examples thereof include deca-9-ynyl and dodeca-4. -Inyl, dodec-11-ynyl, tetradec-5-ynyl, tetradec-6-ynyl, hexadec-7-ynyl, hexadec-3,5-diynyl, hexadec-5,7-diynyl or octadec-9-ynyl Preferably hexadeca-7-ynyl or octadec-9-ynyl, and more preferably octadec-9-ynyl.

In the formula (CL-I), R ¹⁰¹ and R ¹⁰³ are preferably the same linear or branched C10-C24 alkyl, C10-C24 alkenyl or C10-C24 alkynyl, and the same linear It is more preferably a linear or branched C10-C24 alkyl or C10-C24 alkenyl, and even more preferably the same linear C10-C24 alkenyl.

Examples of C1-C3 alkylene include methylene, ethylene, or propylene, preferably methylene or ethylene, and more preferably methylene.

Examples of the C1-C6 alkyl include methyl, ethyl, propyl, isopropyl, cyclopropyl, butyl, isobutyl, sec-butyl, tert-butyl, cyclobutyl, cyclopropylmethyl, pentyl, isopentyl, sec-pentyl, neopentyl, tert- Examples include pentyl, cyclopentyl, hexyl, cyclohexyl, etc., preferably methyl, ethyl, propyl, isopropyl, butyl, isobutyl, sec-butyl, tert-butyl, pentyl, isopentyl, sec-pentyl, tert-pentyl, neopentyl, hexyl, etc. More preferably, methyl, ethyl, propyl, etc. are mentioned.

Examples of C3-C6 alkenyl include allyl, 1-propenyl, butenyl, pentenyl, hexenyl and the like, preferably allyl and the like.

Monoalkylamino and dialkylamino are each one or two identical or different C1-C6 alkyls (as defined above) or amino, methylamino, ethylamino, dimethylamino, diethylamino, pyrrolidinyl, piperidyl or morpholinyl Any amino substituted with C1-C6 alkyl (as defined above) substituted with, for example, methylamino, ethylamino, propylamino, butylamino, pentylamino, hexylamino, dimethylamino, diethylamino, ethylmethylamino Methylpropylamino, butylmethylamino, methylpentylamino, hexylmethylamino, aminoethylamino, aminopropylamino, (aminoethyl) methylamino, bis (aminoethyl) amino, etc., preferably methylamino Bruno, ethylamino, dimethylamino, diethylamino, aminopropyl amino or bis (aminoethyl) amino and the like, more preferably methyl or dimethylamino and the like.

Trialkylammonio includes the same or different C1-C6 alkyl (as defined above), or C1-C6 substituted with amino, methylamino, ethylamino, dimethylamino, diethylamino, pyrrolidinyl, piperidyl or morpholinyl. Any ammonio substituted with alkyl (as defined above) may be used, for example, trimethylammonio, ethyldimethylammonio, diethylmethylammonio, triethylammonio, tripropylammonio, tributylammonio, tripentylammonio, tripentylammonio, Hexylammonio, tris (aminoethyl) ammonio, (aminoethyl) dimethylammonio, bis (aminoethyl) methylammonio and the like can be mentioned, preferably trimethylammonio, triethylammonio, tris (aminoethyl) ammonio (Aminoethyl) dimethyl ammonio or bis (aminoethyl) methyl ammonio, and the like, or more preferably trimethylammonio like.

In the compound (CL-I), the trialkylammonio may form a salt with a pharmaceutically acceptable anion (as defined above).

Alkoxy is substituted with C1-C6 alkyl (as defined above) or C1-C6 alkyl (as defined above) substituted with amino, methylamino, ethylamino, dimethylamino, diethylamino, pyrrolidinyl, piperidyl or morpholinyl. It may be hydroxy, for example, methoxy, ethoxy, propyloxy, butyloxy, pentyloxy, hexyloxy, aminoethoxy or methylaminoethoxy, preferably methoxy, ethoxy, aminoethoxy or methylaminoethoxy, More preferably, methoxy etc. are mentioned.

Monoalkylcarbamoyl and dialkylcarbamoyl are each one or two identical or different C1-C6 alkyls (as defined above) or amino, methylamino, ethylamino, dimethylamino, diethylamino, pyrrolidinyl, piperidyl or morpholinyl Any carbamoyl substituted with C1-C6 alkyl substituted with (as defined above), for example, methylcarbamoyl, ethylcarbamoyl, propylcarbamoyl, butylcarbamoyl, pentylcarbamoyl, hexylcarbamoyl, dimethylcarbamoyl, diethylcarbamoyl, ethyl Methylcarbamoyl, methylpropylcarbamoyl, butylmethylcarbamoyl, methylpentylcarbamoyl, hexylmethylcarbamoyl, aminoethylcarbamoyl, amino Examples include propylcarbamoyl, (aminoethyl) methylcarbamoyl, or bis (aminoethyl) carbamoyl, preferably methylcarbamoyl, ethylcarbamoyl, dimethylcarbamoyl, and the like, more preferably methylcarbamoyl, dimethylcarbamoyl, and the like. .

L ¹⁰¹ and L ¹⁰² are more preferably hydrogen atoms. In this case, R ¹⁰¹ and R ¹⁰² are the same or different and dodecyl, tetradecyl, (Z) -dodec-7-enyl, (Z) -tetradec-7-enyl, (Z) -hexadec-4-enyl. (Z) -hexadec-7-enyl, (E) -hexadeca-7-enyl, (Z) -hexadec-9-enyl, (7Z, 10Z) -hexadec-7,10-dienyl, (7Z, 10Z, 13Z) -hexadeca-7,10,13-trienyl, preferably (Z) -octadeca-9-enyl or (9Z, 12Z) -octadeca-9,12-dienyl, (Z) -tetradec-7- More preferably, they are enyl, (Z) -hexadec-7-enyl, (7Z, 10Z) -hexadeca-7,10-dienyl, or (9Z, 12Z) -octadec-9,12-dienyl. Z) -tetradec-7-enyl, (Z) -hexadeca-7-enyl or (7Z, 10Z) -hexadeca-7,10-dienyl or (9Z, 12Z) -octadeca-9,12-dienyl Further preferred.
In the case where L ¹⁰¹ and L ¹⁰² are hydrogen atoms, X ¹⁰¹ is a hydrogen atom, methyl, pyrrolidin-3-yl, piperidin-3-yl, piperidin-4-yl, or the same or different from 1 to Must be C1-C6 alkyl or C3-C6 alkenyl substituted with 3 amino, monoalkylamino, dialkylamino, trialkylammonio, hydroxy, alkoxy, carbamoyl, monoalkylcarbamoyl, dialkylcarbamoyl, pyrrolidinyl, piperidyl or morpholinyl Is more preferably a hydrogen atom, methyl, or C1-C6 alkyl or C3-C6 alkenyl substituted with 1 to 3 amino, hydroxy or carbamoyl, which are the same or different, and more preferably a hydrogen atom, methyl, etc. More preferably it is.

When L ¹⁰¹ and L ¹⁰² together form a single bond or C1-C3 alkylene, R ¹⁰¹ and R ¹⁰² are the same or different and are tetradecyl, hexadecyl, (Z) -tetradec-9-enyl, (Z) -hexadec-9-enyl, (Z) -octadeca-6-enyl, (Z) -octadeca-9-enyl, (E) -octadeca-9-enyl, (Z) -octadeca-11-enyl, (9Z, 12Z) -octadeca-9,12-dienyl, (9Z, 12Z, 15Z) -octadeca-9,12,15-trienyl, (Z) -icosa-11-enyl, or (11Z, 14Z) -icosa -11,14-dienyl, more preferably (Z) -octadec-9-enyl, or (9Z, 12Z) -octadec-9,12-dienyl, and identically (Z) -octadeca More preferred is -9-enyl or (9Z, 12Z) -octadeca-9,12-dienyl.

When L ¹⁰¹ and L ¹⁰² together form a single bond or C1-C3 alkylene, X ¹⁰¹ is a hydrogen atom, methyl, pyrrolidin-3-yl, piperidin-3-yl, piperidin-4-yl Or C1-C6 alkyl substituted with 1 to 3 amino, monoalkylamino, dialkylamino, trialkylammonio, hydroxy, alkoxy, carbamoyl, monoalkylcarbamoyl, dialkylcarbamoyl, pyrrolidinyl, piperidyl or morpholinyl, the same or different Or a C3-C6 alkenyl, more preferably a hydrogen atom, methyl, or a C1-C6 alkyl or C3-C6 alkenyl substituted with 1 to 3 amino, hydroxy or carbamoyl, which are the same or different. And most preferably a hydrogen atom or methyl.

When L ¹⁰¹ and L ¹⁰² together form a single bond, L ¹⁰³ is —CO— or —CO—O—, preferably —CO—. One. In this case, X ¹⁰¹ is aminomethyl, 1,2-diaminoethyl, 2-aminoethyl, 1,3-aminopropyl, 1,4-diaminobutyric, 1,5 Jiaminopenchiru, 3-aminopropyl, 4 -Aminobutyl or 5-aminopentyl is preferable, and 1,2-diaminoethyl, 1,3-diaminopropyl, 1,4-diaminobutyl or 1,5-diaminopentyl is more preferable. R ¹⁰¹ and R ¹⁰² are the same or different and are tetradecyl, hexadecyl, (Z) -tetradec-9-enyl, (Z) -hexadeca-9-enyl, (Z) -octadec-6-enyl, (Z)- Octadeca-9-enyl, (E) -octadeca-9-enyl, (Z) -octadeca-11-enyl, (9Z, 12Z) -octadeca-9,12-dienyl, (9Z, 12Z, 15Z) -octadeca- Preferably it is 9,12,15-trienyl or (Z) -icosa-11-enyl or (11Z, 14Z) -icosa-11,14-dienyl, (Z) -octadeca-9-enyl or (9Z, More preferably, they are 12Z) -octadeca-9,12-dienyl, and more preferably (Z) -octadeca-9-enyl or (9Z, 12Z) -octadeca-9,12-dienyl.

L ¹⁰³ is more preferably a single bond.

When L ¹⁰³ is a single bond, X ¹⁰¹ is a hydrogen atom, methyl, pyrrolidin-3-yl, piperidin-3-yl, piperidin-4-yl, or the same or different 1 to 3 amino, monoalkylamino , Dialkylamino, trialkylammonio, hydroxy, alkoxy, carbamoyl, monoalkylcarbamoyl, dialkylcarbamoyl, pyrrolidinyl, piperidyl or morpholinyl substituted C1-C6 alkyl or C3-C6 alkenyl, etc., more preferably a hydrogen atom , Methyl, hydroxymethyl, 2-hydroxyethyl, 2,3-dihydroxypropyl, 2-hydroxypropyl, 3-hydroxypropyl, 2-hydroxy-3-methoxypropyl, aminomethyl, 2-aminoethyl, 3-aminopropyl, 4-aminobutyl, 5-aminopentyl, 2- (N, N-dimethylamino) ethyl 3- (N, N-dimethylamino) propyl, 2-carbamoylethyl, 2-dimethylcarbamoylethyl, 1-methylpiperidin-4-yl, and the like, more preferably a hydrogen atom or methyl. preferable.

When L ¹⁰³ is —CO— or —CO—O—, X ¹⁰¹ is pyrrolidin-3-yl, piperidin-3-yl, piperidin-4-yl, or the same or different one to three amino, mono Alkylamino, dialkylamino, trialkylammonio, hydroxy, alkoxy, carbamoyl, monoalkylcarbamoyl, dialkylcarbamoyl, pyrrolidinyl, piperidyl or morpholinyl substituted C1-C6 alkyl or C3-C6 alkenyl, at least of the substituents More preferably, one is amino, monoalkylamino, dialkylamino, trialkylammonio, pyrrolidinyl, piperidyl, morpholinyl or the like, and R ³ is aminomethyl, 1,2-diaminoethyl, 2-aminoethyl, 1 , 3-Diaminopropyl, 3-aminopropyl, 1,4-diaminobutyl, 4-aminobutyl 1,5-diaminopentyl, 5-aminopentyl, (N, N-dimethylamino) methyl, 2- (N, N-dimethylamino) ethyl, 3- (N, N-dimethylamino) propyl, 1-hydroxy- 2-aminoethyl or 1-amino-2-hydroxyethyl is more preferable, such as 1,2-diaminoethyl, 2-aminoethyl, 1,3-diaminopropyl, 3-aminopropyl, 1,4-diamino. Most preferred is butyl, 4-aminobutyl, 1,5-diaminopentyl or 5-aminopentyl.

One of the more preferred embodiments of the present invention is that L ¹⁰³ is a single bond and X ¹⁰¹ is a hydrogen atom. In this case, R ¹⁰¹ and R ¹⁰² are the same or different and dodecyl, tetradecyl, (Z) -dodec-7-enyl, (Z) -tetradec-7-enyl, (Z) -hexadec-4-enyl. (Z) -hexadec-7-enyl, (E) -hexadeca-7-enyl, (Z) -hexadec-9-enyl, (7Z, 10Z) -hexadec-7,10-dienyl, (7Z, 10Z, 13Z) -hexadeca-7,10,13-trienyl, (Z) -octadeca-9-enyl or (9Z, 12Z) -octadeca-9,12-dienyl, preferably the same or different, (Z) More preferred are -tetradec-7-enyl or (7Z, 10Z) -hexadec-7,10-dienyl, and (Z) -tetradec-7-enyl, (Z) -hexadeca-7-enyl or ( 7Z, 10Z) -hexadec-7,10-dienyl is more preferable.

L ¹⁰³ is a single bond and X ¹⁰¹ is methyl is one of the more preferable embodiments of the present invention. In this case, R ¹⁰¹ and R ¹⁰² are the same or different and dodecyl, tetradecyl, (Z) -dodec-7-enyl, (Z) -tetradec-7-enyl, (Z) -hexadec-4-enyl. (Z) -hexadec-7-enyl, (E) -hexadeca-7-enyl, (Z) -hexadec-9-enyl, (7Z, 10Z) -hexadec-7,10-dienyl, (7Z, 10Z, 13Z) -hexadeca-7,10,13-trienyl, (Z) -octadeca-9-enyl or (9Z, 12Z) -octadeca-9,12-dienyl, preferably the same or different, (Z) More preferred are tetradeca-7-enyl, (7Z, 10Z) -hexadec-7,10-dienyl or (9Z, 12Z) -octadeca-9,12-dienyl, and identically (Z) -tetradec-7 More preferred is -enyl, (7Z, 10Z) -hexadec-7,10-dienyl or (9Z, 12Z) -octadeca-9,12-dienyl.

In the definition of each group of formula (CL-II), linear or branched C12-C24 alkyl includes, for example, dodecyl, tridecyl, tetradecyl, 2,6,10-trimethylundecyl, pentadecyl, 3,7, 11-trimethyldodecyl, hexadecyl, heptadecyl, octadecyl, 6,10,14-trimethylpentadecan-2-yl, nonadecyl, 2,6,10,14-tetramethylpentadecyl, icosyl, 3,7,11,15-tetra Examples include methylhexadecyl, henicosyl, docosyl, tricosyl, tetracosyl, etc., preferably dodecyl, tridecyl, tetradecyl, pentadecyl, hexadecyl, heptadecyl, octadecyl, nonadecyl, icosyl, etc., more preferably dodecyl, tridecyl, tetradecyl, pentadecyl , Hexadecyl, heptadecyl, octadecyl, etc. It is.

The linear or branched C12-C24 alkenyl may be a linear or branched C12-C24 alkenyl containing 1 to 3 double bonds, such as (Z) -tridec-8-enyl. , (Z) -tetradec-9-enyl, (Z) -pentadeca-8-enyl, (Z) -hexadeca-9-enyl, (Z) -heptadeca-5-enyl, (Z) -octadeca-6-enyl , (Z) -heptadeca-8-enyl, (Z) -octadeca-9-enyl, (E) -heptadeca-8-enyl, (E) -octadeca-9-enyl, (Z) -heptadeca-10-enyl , (Z) -octadeca-11-enyl, (8Z, 11Z) -heptadeca-8,11-dienyl, (9Z, 12Z) -octadeca-9,12-dienyl, (8Z, 11Z, 14Z) -octadeca-8 , 11,14-trienyl, (9Z, 12Z, 15Z) -octadeca-9,12,15-trienyl, (Z) -nonadec-10-enyl, (Z) -icosa-11-enyl, (10Z, 13Z) -Nonadeca-10,13-dienyl, (11Z, 14Z) -icosa-11,14-dienyl, 2,6,10-trimethylundeca-1,5,9-trienyl, 3,7 , 11-trimethyldodeca-2,6,10-trienyl, 2,6,10,14-tetramethylpentadec-1-enyl or 3,7,11,15-tetramethylhexadec-2-enyl (Z) -pentadec-8-enyl, (Z) -hexadeca-9-enyl, (Z) -heptadeca-5-enyl, (Z) -octadeca-6-enyl, (Z) -heptadeca- 8-enyl, (Z) -octadec-9-enyl, (8Z, 11Z) -heptadeca-8,11-dienyl, (9Z, 12Z) -octadeca-9,12-dienyl, and the like, more preferably ( Z) -heptadeca-8-enyl, (Z) -octadeca-9-enyl, (8Z, 11Z) -heptadeca-8,11-dienyl or (9Z, 12Z) -octadeca-9,12-dienyl .

The linear or branched C12-C24 alkynyl may be a linear or branched C12-C24 alkynyl containing 1 to 3 triple bonds, such as dodeca-11-ynyl, tridec-12- Inyl, pentadec-6-ynyl, hexadec-7-ynyl, pentadec-4,6-diynyl, hexadec-5,7-diynyl, heptadec-8-ynyl, octadec-9-ynyl and the like, preferably pentadec-9 6-Inyl, hexadec-7-ynyl, pentadec-4,6-diynyl, hexadec-5,7-diynyl, heptadec-8-ynyl, octadec-9-ynyl, etc., more preferably heptadeca-8-inyl Or octadec-9-inyl etc. are mentioned.

C1-C3 alkylene, C1-C6 alkyl and C3-C6 alkenyl in the definition of each group of formula (CL-II) are respectively synonymous with those in formula (CL-I).

Monoalkylamino, dialkylamino, trialkylammonio, alkoxy, monoalkylcarbamoyl and dialkylcarbamoyl have the same meanings as those in formula (CL-I), respectively.

R ¹⁰³ and R ¹⁰⁴ are preferably the same linear or branched C12-C24 alkyl, C12-C24 alkenyl or C12-C24 alkynyl, and the same linear or branched C12-C24 alkyl or More preferred is C12-C24 alkenyl.

L ¹⁰⁴ and L ¹⁰⁵ are the same -O -, - more preferably CO-O- or -O-CO-.

When at least one of L ¹⁰⁴ and L ¹⁰⁵ is —O— or —O—CO—, R ¹⁰³ and R ¹⁰⁴ are the same or different and are dodecyl, tetradecyl, hexadecyl, octadecyl, icosyl, docosyl, tetracosyl, (Z) -tetradec-9-enyl, (Z) -hexadeca-9-enyl, (Z) -octadeca-6-enyl, (Z) -octadeca-9-enyl, (E) -octadeca-9-enyl, (Z) -octadeca-11-enyl, (9Z, 12Z) -octadeca-9,12-dienyl, (9Z, 12Z, 15Z) -octadeca-9,12,15-trienyl, (Z) -icosa-11- Enyl, (11Z, 14Z) -icosa-11,14-dienyl, 3,7,11-trimethyldodeca-2,6,10-trienyl or 3,7,11,15-tetramethylhexadec-2-enyl More preferably, tetradecyl, hexadecyl, octadecyl, (Z) -hexadec-9-enyl, (Z) -octadec-6-enyl, (Z) -octadec-9-enyl or (9Z, 12Z) -octadec- 9, More preferred is 12-dienyl.

When at least one of L ¹⁰⁴ and L ¹⁰⁵ is -CO-O-, R ¹⁰³ and R ¹⁰⁴ are each tridecyl, pentadecyl, heptadecyl, nonadecyl, henicosyl, tricosyl, (Z) -tridec-8-enyl, ( (Z) -pentadeca-8-enyl, (Z) -heptadeca-5-enyl, (Z) -heptadeca-8-enyl, (E) -heptadeca-8-enyl, (Z) -heptadeca-10-enyl, ( 8Z, 11Z) -heptadeca-8,11-dienyl, (8Z, 11Z, 14Z) -octadeca-8,11,14-trienyl, (Z) -nonadec-10-enyl, (10Z, 13Z) -nonadec-10 , 13-dienyl, (11Z, 14Z) -icosa-11,14-dienyl, 2,6,10-trimethylundeca-1,5,9-trienyl or 2,6,10,14-tetramethylpentadeca- More preferred is 1-enyl, tridecyl, pentadecyl, heptadecyl, (Z) -pentadec-8-enyl, (Z) -heptadeca-5-enyl, (Z) -heptadeca-8-enyl or (8Z, 11Z ) -Heptadeca-8,11- Further preferably enyl.

More preferably, p ¹⁰¹ and p ¹⁰² are 0 or 1 at the same time.

More preferably, L ¹⁰⁶ and L ¹⁰⁷ together form a single bond or C1-C3 alkylene. When L ¹⁰⁶ and L ¹⁰⁷ together form a single bond or C1-C3 alkylene, X ¹⁰² is a hydrogen atom, methyl, pyrrolidin-2-yl, pyrrolidin-3-yl, piperidin-2-yl , Piperidin-3-yl, piperidin-4-yl, morpholin-2-yl, morpholin-3-yl, or the same or different 1 to 3 amino, monoalkylamino, dialkylamino, trialkylammonio, hydroxy, More preferably it is C1-C6 alkyl or C3-C6 alkenyl substituted with alkoxy, carbamoyl, monoalkylcarbamoyl, dialkylcarbamoyl, pyrrolidinyl, piperidyl or morpholinyl, hydrogen atom, methyl, or the same or different from 1 to 3 C1-C6 alkyl substituted with amino, trialkylammonio, hydroxy or carbamoyl Or more preferably a C3-C6 alkenyl, a hydrogen atom, methyl, 2,3-dihydroxypropyl, 3-hydroxypropyl, aminomethyl, 1,2-diaminoethyl, 2-aminoethyl, 1,3-diamino. Most preferred is propyl, 1,4-diaminobutyl, 1,5-diaminopentyl, 3-aminopropyl, 4-aminobutyl, 5-aminopentyl or 2-carbamoylethyl. Of these substituents, the alkyl moiety in monoalkylamino, dialkylamino, trialkylammonio, alkoxy, monoalkylcarbamoyl and dialkylcarbamoyl has the same meaning as the C1-C4 alkyl. Two or three alkyls in dialkylamino, trialkylammonio and dialkylcarbamoyl may be the same or different.

When L ¹⁰⁶ and L ¹⁰⁷ together form a single bond, L ¹⁰⁸ is preferably —CO— or —CO—O—, preferably —CO—.

When L ¹⁰⁶ and L ¹⁰⁷ together form a single bond, p ¹⁰¹ and p ¹⁰² are preferably the same or different and are 1 to 3.

When L ¹⁰⁶ and L ¹⁰⁷ are hydrogen atoms, X ¹⁰² is a hydrogen atom, methyl, pyrrolidin-2-yl, pyrrolidin-3-yl, piperidin-2-yl, piperidin-3-yl, piperidine-4 -Yl, morpholin-2-yl, morpholin-3-yl or the same or different 1 to 3 amino, monoalkylamino, dialkylamino, trialkylammonio, hydroxy, alkoxy, carbamoyl, monoalkylcarbamoyl, dialkylcarbamoyl, C1-C6 alkyl or C3-C6 alkenyl substituted with pyrrolidinyl, piperidyl or morpholinyl, preferably hydrogen atom, methyl or the same or different from 1 to 3 amino, trialkylammonio, hydroxy or carbamoyl More preferred is a substituted C1-C6 alkyl or C3-C6 alkenyl. Hydrogen atom, methyl, 2,3-dihydroxypropyl, 3-hydroxypropyl, aminomethyl, 1,2-diaminoethyl, 2-aminoethyl, 1,3-diaminopropyl, 1,4-diaminobutyl, 1, More preferred are 5-diaminopentyl, 3-aminopropyl, 4-aminobutyl, 5-aminopentyl, 2-carbamoylethyl and the like. Of these substituents, the alkyl moiety in monoalkylamino, dialkylamino, trialkylammonio, alkoxy, monoalkylcarbamoyl and dialkylcarbamoyl has the same meaning as the C1-C4 alkyl. Two or three alkyls in dialkylamino, trialkylammonio, and dialkylcarbamoyl may be the same or different.

L ¹⁰⁸ is preferably a single bond. When L ¹⁰⁸ is a single bond, L ¹⁰⁴ and L ¹⁰⁵ are preferably —O—.

When L ¹⁰⁸ is a single bond, X ¹⁰² is a hydrogen atom, methyl, pyrrolidin-3-yl, piperidin-3-yl, piperidin-4-yl, or the same or different 1 to 3 amino, monoalkylamino, C1-C6 alkyl or C3-C6 alkenyl substituted with dialkylamino, trialkylammonio, hydroxy, alkoxy, carbamoyl, monoalkylcarbamoyl, dialkylcarbamoyl, pyrrolidinyl, piperidyl or morpholinyl is preferred. , Hydroxymethyl, 2-hydroxyethyl, 2,3-dihydroxypropyl, 2-hydroxypropyl, 3-hydroxypropyl, 2-hydroxy-3-methoxypropyl, aminomethyl, 2-aminoethyl, 3-aminopropyl, 4- Aminobutyl, 5-aminopentyl, 2- (N, N-dimethylamino) ethyl, 3- (N, N-dimethylamino) propyl, 2-carbamoylethyl, 2-dimethylcarbamoylethyl, 1-methylpiperidin-4-yl and the like are more preferable, such as a hydrogen atom, methyl, 2,3-dihydroxypropyl, 3-hydroxypropyl. 2-aminoethyl, 3-aminopropyl, 4-aminobutyl, 5-aminopentyl, 2-carbamoylethyl and the like are more preferable. Of these substituents, the alkyl moiety in monoalkylamino, dialkylamino, trialkylammonio, alkoxy, monoalkylcarbamoyl and dialkylcarbamoyl has the same meaning as the C1-C4 alkyl. Two or three alkyls in dialkylamino, trialkylammonio, and dialkylcarbamoyl may be the same or different.

More preferably, L ¹⁰⁴ and L ¹⁰⁵ are —O—. However, when L ¹⁰⁸ is a single bond and X ¹⁰² is a hydrogen atom, L ¹⁰⁴ and L ¹⁰⁵ are preferably the same —CO—O— or —O—CO—, and —CO—O— More preferably.

When L ¹⁰⁸ is —CO— or —CO—O—, L ¹⁰⁴ and L ¹⁰⁵ are preferably the same —CO—O— or —O—CO—, and are —CO—O—. Is more preferable.

When L ¹⁰⁸ is —CO— or —CO—O—, X ¹⁰² is pyrrolidin-3-yl, piperidin-3-yl, piperidin-4-yl or the same or different 1 to 3 amino, monoalkyl Preferably it is C1-C6 alkyl or C3-C6 alkenyl substituted with amino, dialkylamino, trialkylammonio, hydroxy, alkoxy, carbamoyl, monoalkylcarbamoyl, dialkylcarbamoyl, pyrrolidinyl, piperidyl or morpholinyl. Is preferably amino, monoalkylamino, dialkylamino, trialkylammonio, pyrrolidinyl, piperidyl or morpholinyl, and X ¹⁰² is aminomethyl, 1,2-diaminoethyl, 2-aminoethyl, 1 , 3-Diaminopropyl, 3-aminopropyl, 1,4-diaminobutyl, 4-a Nobutyl, 1,5-diaminopentyl, 5-aminopentyl, (N, N-dimethylamino) methyl, 2- (N, N-dimethylamino) ethyl, 3- (N, N-dimethylamino) propyl or 1- More preferred are amino-2-hydroxyethyl and the like, aminomethyl, 1,2-diaminoethyl, 2-aminoethyl, 1,3-diaminopropyl, 3-aminopropyl, 1,4-diaminobutyl, 4- More preferred are aminobutyl, 1,5-diaminopentyl, 5-aminopentyl and the like. Of these substituents, the alkyl moiety in monoalkylamino, dialkylamino, trialkylammonio, alkoxy, monoalkylcarbamoyl and dialkylcarbamoyl has the same meaning as the C1-C6 alkyl. Two or three alkyls in dialkylamino, trialkylammonio and dialkylcarbamoyl may be the same or different.

L ¹⁰⁴ and L ¹⁰⁵ are preferably the same —CO—O— or —O—CO—, and more preferably —CO—O—.

In the definitions of the groups of formulas (CL-III), (CL-IV) and (CL-V), linear or branched C8-C24 alkyl, C8-C24 alkenyl and C8-C24 alkynyl are the same as defined above. As defined in formulas (I) to (IV), the same groups are preferred.

In the definition of each group of the formulas (CL-III), (CL-IV) and (CL-V), the alkyl moiety in C8-C24 alkyloxyethyl and C8-C24 alkyloxypropyl may be, for example, the linear or Examples thereof include those exemplified for branched C8-C24 alkyl.

Examples of the alkynyl moiety in alkynyloxyethyl and alkynyloxypropyl include those exemplified for the linear or branched C8-C24 alkynyl.

R ¹⁰⁵ and R ¹⁰⁶ are preferably the same or different and are linear or branched C8-C24 alkyl or C8-C24 alkenyl, and are the same or different, linear or branched C8-C24 alkenyl. More preferably, it is more preferably the same or different and linear C8-C24 alkenyl. R ¹⁰⁵ and R ¹⁰⁶ are more preferably the same, and in that case, linear or branched C12-C24 alkyl, C12-C24 alkenyl, or C12-C24 alkynyl is preferable. More preferably, it is a chain C12-C24 alkenyl. Linear or branched C12-C24 alkyl, C12-C24 alkenyl, and C12-C24 alkynyl have the same meanings as those in formula (CL-II), respectively.

R ¹⁰⁵ and R ¹⁰⁶ are preferably the same or different and are linear or branched C8-C24 alkyl or C8-C24 alkenyl, and are the same or different, linear or branched C8-C24 alkenyl. More preferably, it is more preferably the same or different and linear C8-C24 alkenyl. R ¹⁰⁵ and R ¹⁰⁶ are more preferably the same, and in that case, linear or branched C15-C20 alkyl, C15-C20 alkenyl, or C15-C20 alkynyl is preferable. More preferably, it is a chain C15-C20 alkenyl. Linear or branched C15-C20 alkyl, C15-C20 alkenyl, and C15-C20 alkynyl have the same meanings as those in formulas (I) to (IV), respectively, and the same groups are preferable.

When R ¹⁰⁵ and R ¹⁰⁶ are different, R ¹⁰⁵ is linear or branched C15-C20 alkyl, C15-C20 alkenyl or C15-C20 alkynyl, and R ¹⁰⁶ is linear or branched C8. -C12 alkyl is preferred. Here, examples of the linear or branched C8-C12 alkyl include octyl, nonyl, decyl, undecyl, and dodecyl, and preferably octyl, decyl, and dodecyl.

A C15-C20 alkenyl R ¹⁰⁵ is linear, more preferably R ¹⁰⁶ is a straight-chain C8-C12 alkyl, R ¹⁰⁵ is (Z) - octadec-9-enyl or (9Z, 12Z) More preferably, it is -octadeca-9,12-dienyl and R ¹⁰⁶ is octyl, decyl or dodecyl.

When R ¹⁰⁵ and R ¹⁰⁶ are different, R ¹⁰⁵ is linear or branched C8-C24 alkyl, C8-C24 alkenyl or C8-C24 alkynyl, R ¹⁰⁶ is C8-C24 alkyloxyethyl, C8-C24 alkyloxypropyl, C8-C24 alkenyloxyethyl, C8-C24 alkenyloxypropyl, C8-C24 alkynyloxyethyl or C8-C24 alkynyloxypropyl is also preferred. In this case, R ¹⁰⁵ is a straight-chain C8-C24 alkenyl, R ¹⁰⁶ is more preferably a C8-C24 alkenyloxy ethyl, R ¹⁰⁵ is, (Z) - octadec-9-enyl, ( 9Z, 12Z) -octadeca-9,12-dienyl or (11Z, 14Z) -icosa-11,14-dienyl, R ¹⁰⁶ is (Z) -octadec-9-enyloxyethyl, (9Z, 12Z) More preferably, it is -octadeca-9,12-dienyloxyethyl or (11Z, 14Z) -icosa-11,14-dienyloxyethyl, and R ¹⁰⁵ is (9Z, 12Z) -octadeca-9,12 Most preferably, it is -dienyl and R ¹⁰⁶ is (9Z, 12Z) -octadeca-9,12-dienyloxyethyl.

When R ¹⁰⁵ and R ¹⁰⁶ are the same or different linear or branched C8-C24 alkyl or C8-C24 alkenyl, the same or different tetradecyl, hexadecyl, (Z) -tetradec-9-enyl , (Z) -hexadeca-9-enyl, (Z) -octadeca-6-enyl, (Z) -octadeca-9-enyl, (E) -octadeca-9-enyl, (Z) -octadeca-11-enyl , (9Z, 12Z) -octadeca-9,12-dienyl, (9Z, 12Z, 15Z) -octadeca-9,12,15-trienyl, (Z) -icosa-11-enyl, (11Z, 14Z) -icosa -11,14-dienyl or (Z) -docosa-13-enyl, the same or different hexadecyl, (Z) -hexadec-9-enyl, (Z) -octadec-6-enyl, (Z ) -Octadeca-9-enyl, (9Z, 12Z) -octadeca-9,12-dienyl, (Z) -icosa-11-enyl or (11Z, 14Z) -icosa-11,14-dienyl Preferably the same or different More preferred are (Z) -octadeca-9-enyl, (9Z, 12Z) -octadeca-9,12-dienyl or (11Z, 14Z) -icosa-11,14-dienyl, identically (9Z, 12Z Most preferred is) -octadeca-9,12-dienyl.

R ¹⁰⁷ has the same meaning as the R ^105, R ¹⁰⁷ is the same group as R ¹⁰⁵ is preferred. R ¹⁰⁸ is linear C8-C24 alkyloxyethyl, C8-C24 alkyloxypropyl, C8-C24 alkenyloxyethyl, C8-C24 alkenyloxypropyl, C8-C24 alkynyloxyethyl, C8-C24 alkynyloxypropyl, C8 -C24 alkyloxyethoxyethyl, C8-C24 alkenyloxyethoxyethyl or C8-C24 alkynyloxyethoxyethyl is preferred, and linear C8-C24 alkyloxyethyl, C8-C24 alkenyloxyethyl, C8-C24 alkynyloxyethyl is preferred. More preferred. Most preferably, R ¹⁰⁷ is linear C15-C20 alkenyl and R ¹⁰⁸ is C8-C24 alkenyloxyethyl.

R ¹⁰⁹ and R ¹¹⁰ are synonymous with R ¹⁰⁵ and R ¹⁰⁶ , respectively, and the same groups as R ¹⁰⁹ and R ¹¹⁰ are preferred. However, R ¹⁰⁹ and R ¹¹⁰ are preferably the same linear or branched C15-C20 alkyl, C15-C20 alkenyl or C15-C20 alkynyl, and the same (9Z, 12Z) -octadeca-9, More preferred is 12-dienyl.

The C1-C3 alkyl in X ¹⁰³ and X ¹⁰⁴ includes, for example, methyl, ethyl, propyl, isopropyl or cyclopropyl, preferably methyl or ethyl, and more preferably methyl.

Examples of the C2-C8 alkylene formed by combining X ¹⁰³ and X ¹⁰⁴ include ethylene, propylene, butylene, pentylene, hexylene, heptylene, octylene, and the like, preferably butylene, pentylene, hexylene, and the like. More preferred is hexylene.

Examples of the C2-C8 alkylene formed by X ¹⁰³ together with L ¹¹¹ include ethylene, propylene, butylene, pentylene, hexylene, heptylene, octylene, and the like, and preferably propylene, butylene, pentylene, and the like. More preferably, propylene or butylene is used, and propylene is more preferably used.

X ¹⁰³ and X ¹⁰⁴ are the same or different and are methyl or ethyl, together form butylene, pentylene or hexylene, or X ¹⁰³ together with L ¹¹¹ form ethylene, propylene or butylene It is preferable to do. X ¹⁰³ and X ¹⁰⁴ are the same or different and are preferably methyl or ethyl, or together, form butylene, pentylene or hexylene, and X ¹⁰³ together with L ¹¹¹ is ethylene, propylene or It is also preferred that it forms butylene and X ¹⁰⁴ is methyl or ethyl. And it is more preferred that X ¹⁰³ and X ¹⁰⁴ are identically methyl or together form hexylene, X ¹⁰³ together with L ¹¹¹ forms propylene or butylene, and X ¹⁰⁴ is methyl. It is further more preferable.

In L ^111, C1-C6 alkyl, C3-C6 alkenyl, monoalkylamino, alkoxy, mono- alkylcarbamoyl and dialkylcarbamoyl have the same meanings as those in each of the formulas (CL-I).

L ¹¹¹ is a hydrogen atom, C1-C6 alkyl, amino, monoalkylamino, hydroxy, alkoxy or C1-C6 alkyl substituted with 1 to 3 amino, monoalkylamino, or hydroxy or alkoxy, the same or different. Or preferably together with X ¹⁰³ to form C2-C6 alkylene, substituted with 1 to 3 amino or hydroxy atoms, hydrogen atom, methyl, amino, methylamino, hydroxy, methoxy, or the same or different More preferably, together with X ¹⁰³ to form ethylene, propylene or butylene, which is a hydrogen atom, C1-C3 alkyl, or hydroxy, or together with X ¹⁰³ More preferably it forms propylene or butylene, together with a hydrogen atom or X ¹⁰³ Most preferably, propylene is formed.

In L ¹⁰⁹ and L ^110, as the C1-C6 alkylene, such as methylene, ethylene, propylene, butylene, etc. pentylene or hexylene and the like, preferably methylene or ethylene and the like.

L ¹⁰⁹ is preferably methylene, ethylene, propylene, or the like, more preferably methylene, ethylene, or the like. L ¹¹⁰ is preferably a single bond, methylene, ethylene, or the like, and is a single bond, methylene, or the like. More preferably. The sum of the carbon numbers of L ¹⁰⁹ and L ¹¹⁰ is preferably 1 to 3, and more preferably 2. In any of these cases, X ¹⁰³ and X ¹⁰⁴ are the same or different, such as methyl or ethyl, and L ¹¹¹ is a hydrogen atom, methyl, amino, methylamino, hydroxy, methoxy, or the same or different 1 ˜3 amino or hydroxy substituted methyls or the like, or X ¹⁰³ and X ¹⁰⁴ together form pentylene, hexylene or heptylene, etc., and L ¹¹¹ is a hydrogen atom, methyl, amino, methylamino, X ¹⁰³ and L ¹¹¹ together form propylene, butylene, pentylene or the like, such as hydroxy, methoxy or methyl substituted with 1 to 3 amino or hydroxy identically or differently, or X ¹⁰⁴ preferably is methyl or ethyl and the like, X ¹⁰³ and X ¹⁰⁴ is methyl, or L ¹¹¹ is a hydrogen atom, X ¹⁰³ and X ¹⁰⁴ are together It forms a pentylene or hexylene, L ¹¹¹ is hydrogen atom or X ¹⁰³ and L ^111, form a propylene together, X ¹⁰⁴ is more preferably methyl and the like.
The C1-C4 alkyl in the X ^105, for example methyl, ethyl, propyl, isopropyl, butyl, include cyclobutyl, etc., preferably methyl. X ¹⁰⁵ is more preferably a hydrogen atom.

In the definition of each group of the formula (CL-V), the C1-C3 alkyl in X ^{105 ′} includes, for example, methyl, ethyl, propyl, isopropyl, cyclopropyl, etc., preferably methyl, ethyl, isopropyl, etc. More preferably, methyl or ethyl is exemplified. X ^{105 ′} is more preferably a hydrogen atom or methyl, and most preferably a hydrogen atom.

In L ^112, as the C1-C3 alkylene, e.g., methylene, ethylene or propylene and the like, preferably methylene or ethylene and the like.

In the definition of each group of formula (CL-VI) and formula (CL-VII), linear or branched optionally substituted C8-C24 alkyl, C8-C24 alkenyl and C8-C24 alkynyl are respectively It has the same meaning as in the above formulas (I) to (V ″).

Examples of the C1-C4 alkyl in the optionally substituted C1-C4 alkyl in R ¹¹⁵ of the formula (CL-VII) include, for example, methyl, ethyl, propyl, isopropyl, cyclopropyl, butyl, isobutyl, sec-butyl, tert -Butyl, cyclobutyl, cyclopropylmethyl, etc. are mentioned, Preferably methyl, ethyl etc. are mentioned, More preferably, methyl is mentioned.
The alkyl part of C1-C4 alkoxy which may be substituted has the same meaning as the C1-C4 alkyl.
Examples of the substituent in the optionally substituted C1-C4 alkyl include amino, monoalkylamino, dialkylamino, pyrrolidin-2-yl, pyrrolidin-3-yl, piperidin-2-yl, piperidin-3-yl, and piperidine 4-yl, morpholin-2-yl, morpholin-3-yl, hydroxy, alkoxy, alkoxycarbonyl, hydroxycarbonyl, carbamoyl, monoalkylcarbamoyl, dialkylcarbamoyl, nitro, cyano, fluoro, chloro, bromo and the like. Of these substituents, the alkyl moiety in monoalkylamino, dialkylamino, alkoxy, alkoxycarbonyl, monoalkylcarbamoyl and dialkylcarbamoyl has the same meaning as the C1-C4 alkyl. Two alkyls in dialkylamino and dialkylcarbamoyl may be the same or different.

Examples of the acyl in the optionally substituted C1-C4 acyloxy include formyl, acetyl, propanoyl, 2-methylpropanoyl, cyclopropanoyl, butanoyl, and preferably acetyl and the like.
Examples of the substituent in C1-C4 acyloxy which may be substituted include amino, monoalkylamino, dialkylamino, pyrrolidin-2-yl, pyrrolidin-3-yl, piperidin-2-yl, piperidin-3-yl and piperidine 4-yl, morpholin-2-yl, morpholin-3-yl, hydroxy, alkoxy, alkoxycarbonyl, hydroxycarbonyl, carbamoyl, monoalkylcarbamoyl, dialkylcarbamoyl, nitro, cyano, fluoro, chloro, bromo and the like. Of these substituents, the alkyl moiety in monoalkylamino, dialkylamino, alkoxy, alkoxycarbonyl, monoalkylcarbamoyl and dialkylcarbamoyl has the same meaning as the C1-C4 alkyl. Two alkyls in dialkylamino and dialkylcarbamoyl may be the same or different.

In the formula (CL-VI), R ¹¹¹ and R ¹¹² are preferably the same linear or branched C8-C24 alkyl, C8-C24 alkenyl or C8-C24 alkynyl, and the same linear or branched More preferably, it is C8-C24 alkyl or C8-C24 alkenyl.

R ¹¹¹ and R ¹¹² are the same or different and are octyl, decyl, dodecyl, tetradecyl, hexadecyl, octadecyl, icosyl, docosyl, tetracosyl, (Z) -tetradec-9-enyl, (Z) -hexadec-9-enyl, ( (Z) -octadeca-6-enyl, (Z) -octadeca-9-enyl, (E) -octadeca-9-enyl, (Z) -octadeca-11-enyl, (9Z, 12Z) -octadeca-9,12 -Dienyl, (9Z, 12Z, 15Z) -octadeca-9,12,15-trienyl, (Z) -icosa-11-enyl, (11Z, 14Z) -icosa-11,14-dienyl, 3,7,11 -Trimethyldodeca-2,6,10-trienyl or 3,7,11,15-tetramethylhexadec-2-enyl or the like, and the same or different, dodecyl, tetradecyl, (Z) -hexadec-9 -Enyl, (Z) -octadeca-6-enyl, (Z) -octadeca-9-enyl or (9Z, 12Z) -octadeca-9,12-dienyl are more preferred and identical. More preferred are (Z) -hexadec-9-enyl, (Z) -octadeca-6-enyl, (Z) -octadeca-9-enyl, (9Z, 12Z) -octadeca-9,12-dienyl, and the like. .

X ¹⁰⁶ and X ¹⁰⁷ are preferably the same or different and are preferably methyl or ethyl, more preferably methyl.

Examples of the C2-C8 alkylene formed by combining X ¹⁰⁶ and X ¹⁰⁷ include ethylene, propylene, butylene, pentylene, hexylene, heptylene, octylene, and the like, preferably butylene, pentylene, hexylene, and the like. More preferred is butylene or pentylene.

X ¹⁰⁶ and X ¹⁰⁷ are preferably the same methyl or taken together to form butylene, pentylene or hexylene.

p ¹⁰³ and ¹⁰⁴ are preferably 0 at the same time, and p ¹⁰⁵ is preferably 1.

L ¹¹³ and L ¹¹⁴ are preferably O at the same time.

In the formula (CL-VII), R ¹¹³ and R ¹¹⁴ are preferably the same linear or branched C8-C24 alkyl, C8-C24 alkenyl or C8-C24 alkynyl, and the same linear or More preferred are branched C8-C24 alkyl or C8-C24 alkenyl.

C1-C3 alkyl and C2-C8 alkylene in X ¹⁰⁹ and X ¹¹⁰ are as defined as in each of the formulas (CL-VI).

R ¹¹⁵ is preferably a hydrogen atom, hydroxy, methyl, methoxy or the like, more preferably a hydrogen atom or hydroxy, and even more preferably a hydrogen atom.

L ¹¹⁵ is preferably —O—CO— or —NH—CO—. In this case, p ¹⁰⁶ is 0 or 1, and p ¹⁰⁷ is preferably an integer of 1 to 3, more preferably p ¹⁰⁶ is 0 and p ¹⁰⁷ is 1 or 3.

L ¹¹⁵ is the case of -CO-O-, p ¹⁰⁶ is 0, it is preferable that p ¹⁰⁷ is an integer of 2 ~ 4, p ¹⁰⁶ is 0, more that p ¹⁰⁷ is 3 preferable.

L ¹¹⁵ is the case of -CO-NH-, p ¹⁰⁶ is 0, it is preferable that p ¹⁰⁷ is an integer of 2 ~ 4, p ¹⁰⁶ is 0, more that p ¹⁰⁷ is 3 preferable.

The definition of each group of formula (CL-VIII) to formula (CL-XVII) may be synonymous with that in formulas (I) to (V ″). CL-VII) may be synonymous.

In addition, each group in the formula (CL-VIII) is in WO 2016/002753, each group in the formula (CL-X) is in WO 2009/129385, each group in the formula (CL-XI) is In International Publication No.2013 / 149140, each group in formula (CL-XII) is in International Publication No.2009 / 129395, each group in formula (CL-XIII) is in International Publication No.2013 / 059496, -XIV) in International Publication No. 2011/149733, each group in Formula (CL-XV) in International Publication No.2011 / 153493, each group in Formula (CL-XVI) in International Publication No. Each group in the formula (CL-XVII) in No. 074085 may be a preferred embodiment of each group described correspondingly in International Publication No. 2013/064911.

L ¹¹⁸ and L ¹¹⁹ in formula (CL-IX) are the same or different and are preferably linear or branched C8-C24 alkylene or C8-C24 alkenylene, more preferably linear or branched C8. -C20 alkylene or C8-C20 alkenylene.

Wherein C1-C6 alkyl (CL-X) in X ¹¹⁷ and X ^118, heterocyclyl or polyamines, halogen atom, R ', OR', SR 1 is selected ', CN, CO ₂ R' or from CONR _'2 It may be substituted with up to 3 substituents.
X ¹¹⁷ and X ¹¹⁸ in formula (CL-X) contain 1 or 2 additional heteroatoms selected from N, O and S in addition to the nitrogen to which they are attached The monocyclic heterocycle is a halogen atom, R ′, OR ′, SR ′, CN, CO ₂ R ′ or CONR ′. It may be substituted with 1 to 3 substituents selected from ₂ .
Here, R ′ is a hydrogen atom or C1-C6 alkyl, and the C1-C6 alkyl as R ′ may be substituted with a halogen atom or OH.

R ¹²⁰ and R ¹²¹ in formula (CL-X) are the same or different and are preferably linear or branched C4-C24 alkyl or C4-C24 alkenyl, more preferably linear or branched C4- C20 alkyl or C4-C20 alkenyl.
C4-C24 alkyl or C4-C24 alkenyl may be substituted with one or more substituents selected from a halogen atom, R ′, OR ′, SR ′, CN, CO ₂ R ′ or CONR ′ _2. .
Here, R ′ is a hydrogen atom or C1-C6 alkyl, and the C1-C6 alkyl as R ′ may be substituted with a halogen atom or OH.

When X ¹¹⁹ and X ¹²⁰ in formula (CL-XI) are linear or branched optionally substituted C6-C20 acyl, the structure other than the carbonyl group in C6-C20 acyl is C5-C19 alkyl. , C5-C19 alkenyl or C5-C19 alkynyl.

R ¹²⁴ and R ¹²⁵ in formula (CL-XII) are the same or different and are preferably linear or branched C8-C24 alkyl or C8-C24 alkenyl, more preferably linear or branched C14. -C20 alkyl or C14-C20 alkenyl.

X ¹²⁵ and X ¹²⁶ C1-C6 alkyl, heterocyclyl or polyamine in the formula (CL-XIV) are selected from halogen atoms, R ′, OR ′, SR ′, CN, CO ₂ R ′ or CONR ′ ₂ It may be substituted with up to 3 substituents.
X ¹²⁵ and X ¹²⁶ in formula (CL-XIV) contain, in addition to the nitrogen to which they are attached, one or two additional heteroatoms selected from N, O and S. In the case of forming a 4 to 7-membered monocyclic hetero ring which may be optionally substituted, the monocyclic hetero ring is a halogen atom, R ′, OR ′, SR ′, CN, CO ₂ R ′ or CONR ′ ₂ It may be substituted with 1 to 3 substituents selected from
Here, R ′ is a hydrogen atom or C1-C6 alkyl, and the C1-C6 alkyl as R ′ may be substituted with a halogen atom or OH.

R ¹²⁸ and R ¹²⁹ in formula (CL-XIV) are the same or different and are preferably linear or branched C4-C24 alkyl or C4-C24 alkenyl, more preferably linear or branched C4- C20 alkyl or C4-C20 alkenyl.
C4-C24 alkyl or C4-C24 alkenyl may be substituted with one or more substituents selected from a halogen atom, R ′, OR ′, SR ′, CN, CO ₂ R ′ or CONR ′ _2. .
Here, R ′ is a hydrogen atom or C1-C6 alkyl, and the C1-C6 alkyl as R ′ may be substituted with a halogen atom or OH.

Specific examples of lipid B used in the present invention are shown in Tables 1 to 10 below, but lipid B of the present invention is not limited thereto.

Next, the method for producing lipid A of the present invention will be described. In addition, in the production method shown below, when a defined group changes under the conditions of the production method or is inappropriate for carrying out the production method, introduction and removal of a protective group commonly used in organic synthetic chemistry Methods [e.g., Methods described in Protective Groups in Organic Synthesis, third edition, by TWGreene, John Wiley & Sons Inc. (1999)], etc. Can be used to obtain the target compound. Further, the order of reaction steps such as introduction of substituents can be changed as necessary.

In addition, etherification (4th edition, Experimental Chemistry Course 20, Synthesis of Organic Compounds II, 4th edition, p.187, Maruzen (1992), etc.), amination (4th edition) Edition Experimental Chemistry Lecture 20 “Synthesis of Organic Compounds II”, 4th Edition, p.279, Maruzen (1992), etc., Esterification (4th Edition Experimental Chemistry Course 22 “Synthesis of Organic Compounds IV”, 4th Edition, p. .43, Maruzen (1992), etc.), amidation (4th edition, Experimental Chemistry Lecture 22, Synthesis of Organic Compounds IV, 4th edition, p.137, Maruzen (1992), etc.) Can also be carried out using general reaction conditions described in the existing literature.

Hereinafter, compound (I) can be obtained by any of the synthetic pathways 1 and 2, or a method according to these methods.

Compound (I) can be obtained from ammonia according to Synthesis Route 1.

(In the formula, Ms represents a methanesulfonyl group, and other groups are as defined above.)

Compound 2 can be obtained by reacting ammonia and compound 1 in a solvent (for example, a polar solvent such as tetrahydrofuran or methanol) at a high temperature (for example, 80 ° C. or more).

Compound 4 is obtained by reacting Compound 2 and Compound 3 at a high temperature (for example, 100 ° C. or higher) in the presence of a base (for example, an inorganic base such as sodium hydroxide). Although a solvent is not particularly required, a high boiling point solvent (for example, a polar solvent such as ethylene glycol) may be used in some cases.

Compound 6 is obtained by reacting Compound 4 and Compound 5 at a high temperature (for example, 100 ° C. or higher) in the presence of a base (for example, an inorganic base such as sodium hydroxide). Although a solvent is not particularly required, a high boiling point solvent (for example, a polar solvent such as ethylene glycol) may be used in some cases.

In each of the above three heating reactions, a microwave reactor can also be suitably used. Further, instead of Compound 1, Compound 3 and Compound 5, a corresponding halide such as bromide or iodide can be used.

Compound 4 in which R ¹ -L ¹ and R ² -L ² are the same can also be obtained by using an excess amount of Compound 1 from ammonia. Compound 6 in which R ² -L ² and R ³ -L ³ are the same can also be obtained by using an excess amount of compound 3 from compound 2. Furthermore, compound 6 in which R ¹ -L ² , R ² -L ² and R ³ -L ³ are the same can also be obtained by using an excess amount of compound 1 from ammonia.

Compound (I) is obtained by reacting Compound 6 and Compound 7 in the presence or absence of a solvent (eg, a halogen-based solvent such as chloroform) at room temperature or high temperature (eg, 100 ° C. or higher). . In addition, for example, by treating (I) with an appropriate anion exchange resin, the anion A ¹ of the compound (I) can be converted into another anion.

Compounds used for the reaction such as Compound 1, Compound 3, Compound 5 and Compound 7 are commercially available products, or the methods described in Examples or modifications thereof, or methods known in the literature (for example, “No. 5 Version Experimental Chemistry Lecture 13 “Synthesis of Organic Compounds I”, 5th Edition, p. 374, Maruzen (2005), etc.) or a method similar thereto.

Compound 1 can also be obtained by treating the corresponding R ¹ -L ¹ -OH with mesylic anhydride or mesylic chloride.

Further among the compound R ^{^¹} -L ¹ -OH, L ¹ is ^{^{^{-Z 1 - (CY 1 Y 2}}} ) p1 - what is (each group as defined above) ^{^{is, R 1 -OMs, R 1 -OH}} , R ¹ -NY ^7A -H (Y ^7A is as defined above) or R ¹ -CO ₂ H and HO- (CY ¹ Y ² ) _p1 -O-PRO ¹ , MsO- (CY ¹ Y ² ) _p1 -O-PRO ¹ , HO ₂ C- (CY ¹ Y ² ) _p1 -O-PRO ¹ or H-NY ^7A- (CY ¹ Y ² ) _p1 -O-PRO ¹ (where PRO ¹ is silyl Any one of the protecting groups (for example, triethylsilyl (TES), tert-butyldimethylsilyl (TBS), tert-butyldiphenylsilyl (TBDPS), etc.)) and a strong base such as sodium hydride Etherification, amination (eg, substitution reaction), esterification (eg, using a condensing agent such as 1-ethyl-3- (3-dimethylaminopropyl) carbodiimide hydrochloride or the like), or (eg, By performing any of the reactions of amidation (using a similar condensing agent) followed by deprotection Rukoto can.
Also, among compounds R ¹ -L ¹ -OH, L ¹ is -Z ^2- (CY ³ Y ⁴ ) _p2 -Z ^3- (CY ⁵ Y ⁶ ) _p3- (each group is as defined above) Similarly, it can be obtained by applying a known reaction one or more times using a reaction substrate corresponding to the target compound.

Compounds 3 and 5 can be prepared in the same manner as Compound 1.

Compound (Ia) can be obtained from Compound 8 according to Synthesis Route 2.

(Wherein M ¹ to M ³ are the same or different-(CY ¹ Y ² ) _p1- or-(CY ³ Y ⁴ ) _p2 -Z ^3- (CY ⁵ Y ⁶ ) _p3- (wherein each group Are as defined above), and the other groups are as defined above.

Compound 8 and Compound 9 are mixed with a base (for example, an organic base such as triethylamine) and a condensing agent (for example, 1-ethyl-3- (3-dimethylamino) in a solvent (for example, a halogen-based solvent such as chloroform). Propyl) carbodiimide hydrochloride or O- (7-aza-1H-benzotriazol-1-yl) -N, N, N ', N'-tetramethyluronium hexafluorophosphate etc.) and activators Treatment with (an activator such as N, N-dimethylaminopyridine) gives compound 10.

Compound 12 is obtained by esterifying Compound 10 and Compound 11 in the same manner as described above.

Further, Compound 14 is obtained by esterifying Compound 12 and Compound 13 in the same manner as described above.

Compound 12 in which R ¹ and R ² are the same can also be obtained by using an excess amount of Compound 9 from Compound 8. Compound 14 in which R ² and R ³ are the same can also be obtained by using an excess amount of compound 11 from compound 10. Further, Compound 14 in which R ¹ , R ² and R ³ are the same can also be obtained by using an excess amount of Compound 9 from Compound 8.

Compound (Ia) is obtained by reacting Compound 14 and Compound 15 in the presence or absence of a solvent (eg, a halogen-based solvent such as chloroform) at room temperature or high temperature (eg, 100 ° C. or higher). . For example, by treating (Ia) with an appropriate anion exchange resin, the anion A ¹ of the compound (Ia) can be converted to another anion.

Compounds used in the reaction such as Compound 8, Compound 9, Compound 11, Compound 14, and Compound 15 are commercially available products, or the methods described in Examples or equivalent methods, or methods known from the literature (for example, "Fifth Edition Experimental Chemistry Course 14, Synthesis of Organic Compounds II", Fifth Edition, p.1, Maruzen (2005), "Fourth Edition Experimental Chemistry Course 22, Organic Compound Synthesis IV", Fourth Edition, p. 1, the method described in Maruzen (1992), etc.) or a method similar thereto.

Hereinafter, the compound (II) can be obtained by any one of the synthesis routes 3 to 16, or a method according to these methods.

Compound (IIa) can be obtained from Compound 15 according to Synthesis Route 3.

(In the formula, M ⁷ is not present and M ⁴ is-(CY ⁸ Y ⁹ ) _p4- , or M ⁷ is not present and M ⁴ is-(CY ¹⁰ Y ¹¹ ) _p5 -Z ^6- (CY ¹² Y ¹³ ) _p6- or M ⁷ is -Z ^5- (CY ¹⁰ Y ¹¹ ) _p5- and M ⁴ _is- (CY ¹² Y ¹³ ) _p6- (wherein each group is as defined above) M ⁸ is not present and M ⁵ is-(CY ⁸ Y ⁹ ) _p4- or M ⁸ is not present and M ⁵ is-(CY ¹⁰ Y ¹¹ ) _p5 -Z ⁶ -(CY ¹² Y ¹³ ) _p6- or M ⁸ is -Z ^5- (CY ¹⁰ Y ¹¹ ) _p5- and M ⁵ _is- (CY ¹² Y ¹³ ) _p6- , and M ⁹ M ⁶ is-(CY ⁸ Y ⁹ ) _p4- or M ⁹ is not present and M ⁶ is-(CY ¹⁰ Y ¹¹ ) _p5 -Z ^6- (CY ¹² Y ¹³ ) _p6- Or M ⁹ is —Z ⁵ — (CY ¹⁰ Y ¹¹ ) _p5 — and M ⁶ is — (CY ¹² Y ¹³ ) _p6 —, and the other groups are as defined above.)

Compound 22 is obtained by sequentially reacting Compound 16, Compound 17, Compound 19, and Compound 21 under the same reaction conditions as in the esterification reaction of Compound 8 and Compound 9 in Synthesis Route 2.

Compound (IIa) is reacted with Compound 22 and Compound 23 by applying the same conditions as those for the synthesis of Compound (Ia) by the reaction of Compound 14 and Compound 15 in Synthesis Route 2. can get. In addition, for example, by treating (IIa) with a suitable anion exchange resin, the anion A ² of the compound (IIa) can be converted into another anion.

Compounds used for the reaction such as Compound 16, Compound 17, Compound 19, Compound 21 and Compound 23 are commercially available products, or the methods described in Examples or equivalent methods, or methods known from the literature (for example, "Fifth Edition Experimental Chemistry Course 14, Synthesis of Organic Compounds II", Fifth Edition, p.1, Maruzen (2005), "Fourth Edition Experimental Chemistry Course 22, Organic Compound Synthesis IV", Fourth Edition, p. 1, the method described in Maruzen (1992), etc.) or a method similar thereto.

Compound 16 can also be obtained by the methods of Synthesis Routes 11 to 15 described later.

Among compounds 17, those in which M ⁷ is -Z ^5- (CY ¹⁰ Y ¹¹ ) _p5- are R ⁴ -OMs, R ⁴ -OH, R ⁴ -NY ^14A -H (Y ^14A is as defined above) Or any one of R ⁴ -CO ₂ H and HO- (CY ¹⁰ Y ¹¹ ) _p5 -CO-O-PRO ² , MsO- (CY ¹⁰ Y ¹¹ ) _p5 -CO-O-PRO ² , HO ₂ C -(CY ¹⁰ Y ¹¹ ) _p5 -CO-O-PRO ² or H-NY ^14A- (CY ¹⁰ Y ¹¹ ) _p5 -CO-O-PRO ² (where PRO ² is a protecting group for the carboxylic acid ( (Eg, methyl, tert-butyl, benzyl, etc.)) and etherification (eg, using a strong base such as sodium hydride), amination (eg, substitution reaction), (eg, 1- After the reaction, either esterification (using a condensing agent such as ethyl-3- (3-dimethylaminopropyl) carbodiimide hydrochloride) or amidation (eg using a similar condensing agent) is performed. It can be obtained by protecting.

Compounds 19 and 21 can be prepared in the same manner as Compound 17.

In the synthesis route 3, X ⁴ can also be introduced first. That is, compound (IIa) can also be obtained by first allowing compound 23 to act on compound 16 and then performing esterification sequentially with compounds 17, 19 and 21.

Compound (IIb) can be obtained from Compound 16 according to Synthesis Route 4.

(Wherein each group is as defined above)

Compound 16 and Compound 24 in a solvent (eg, an aprotic solvent such as tetrahydrofuran or toluene) in the presence of a base (eg, an inorganic base such as sodium hydride) at a high temperature (eg, 100 ° C. or higher) Compound 25 can be obtained by reaction.

Compound 27 is obtained by etherifying Compound 25 and Compound 26 in the same manner as described above.

Compound 29 is obtained by etherifying Compound 27 and Compound 28 in the same manner as described above.

In each of the above three heating reactions, a microwave reactor can also be suitably used. Further, bromide or iodide corresponding to these can be used in place of compound 24, soot compound 26 and compound 28.

Compound 27 in which R ⁴ -M ⁷ and R ⁵ -M ⁸ are the same can also be obtained by using an excess amount of compound 24 from compound 16. Compound 29 in which R ⁵ -M ⁸ and R ⁶ -M ⁹ are the same can also be obtained by using an excess amount of compound 26 from compound 25. Further, Compound 29 in which R ⁴ -M ⁷ , R ⁵ -M ⁸ and R ⁶ -M ⁹ are the same can also be obtained by using an excess amount of Compound 24 from Compound 16.

Compound (IIb) is obtained by reacting Compound 29 and Compound 23 in the presence or absence of a solvent (for example, a halogen-based solvent such as chloroform) at room temperature or high temperature (for example, 100 ° C. or higher). . In addition, for example, by treating (IIb) with an appropriate anion exchange resin, the anion A ² of the compound (IIb) can be converted to another anion.

Compounds used for the reaction such as Compound 16, Compound 24, Compound 26, Compound 28, and Compound 23 are commercially available products, or the methods described in Examples or a method analogous thereto, or methods known from the literature (for example, "Fifth Edition Experimental Chemistry Course 14, Synthesis of Organic Compounds II", Fifth Edition, p.1, Maruzen (2005), "Fifth Edition Experimental Chemistry Course 13, Synthesis of Organic Compounds I", Fifth Edition, p. 374, Maruzen (2005), etc.) or a similar method.

Compound 24 can also be obtained by treating the corresponding R ⁴ -M ⁷ -OH with mesylic anhydride or mesylic chloride.

Further among the compound R ⁴ -M ^⁷ -OH, M ⁷ is ^{^{^{-Z 5 - (CY 10 Y 11}}} ) p5 - ones (each group is as defined above) in ^{^{which, R 4 -OMs, R 4 -OH}} , R ⁴ -NY ^14A -H (Y ^14A is as defined above) or R ⁴ -CO ₂ H, HO- (CY ¹⁰ Y ¹¹ ) _p5 -O-PRO ¹ , MsO- (CY ¹⁰ Y ¹¹ ) _p5 -O-PRO ¹ , HO ₂ C- (CY ¹⁰ Y ¹¹ ) _p5 -O-PRO ¹ or H-NY ^14A- (CY ¹⁰ Y ¹¹ ) _p5 -O-PRO ¹ (each group is as defined above) Any one of these can be obtained by deprotecting after etherification, amination, esterification or amidation reaction.

Compounds 26 and 28 can be prepared in the same manner as Compound 24

As shown in Synthesis Route 5, Compound (IIc) can be obtained from Compound 25 obtained in Synthesis Route 4 by appropriately combining each reaction such as esterification in Synthesis Route 3. Further, as shown in Synthesis Route 5, Compound (IId) can be obtained from Compound 27 obtained by Synthesis Route 4 by appropriately combining each reaction such as esterification in Synthesis Route 3.

(Wherein each group is as defined above)

Compound (IIe) can be obtained from Compound 30 according to Synthesis Route 6.

(Wherein M ¹⁰ , M ¹¹ and M ¹² are each independently O or NY ^14A , and the other groups are as defined above)

Compound 33 is obtained by subjecting compound 30 to compound 17, compound 19, and compound 21 in this order by esterification or amidation.

Compound (IIe) is reacted with Compound 33 and Compound 23 by applying the same conditions as those for the synthesis of Compound (Ia) by the reaction of Compound 14 and Compound 15 in Synthesis Route 2. can get. In addition, for example, by treating (IIe) with an appropriate anion exchange resin, the anion A ² of the compound (IIe) can be converted into another anion.

Compound 30 can also be obtained by the methods of Synthesis Routes 11 to 15 described later.

Compound (IIf) can be obtained from Compound 34 according to Synthesis Route 7.

(Wherein each group has the same meaning as above)

Compound 35 is obtained by protecting compound 34 with an appropriate protecting group.

Compound 36 is obtained by reacting Compound 35 with Compound 23 under the same conditions as those used in the synthesis of Compound (Ia) by the reaction of Compound 14 and Compound 15 in Synthesis Route 2. Can be obtained by deprotection under appropriate conditions.

Compound (IIf) is obtained by subjecting compound 36, compound 37, compound 39 and compound 41 to an esterification or amidation reaction in this order. In addition, for example, the anion A ² of the compound (IIf) can be converted to another anion by treating the compound (IIf) with an appropriate anion exchange resin.

Compounds 34, 35 and 36 can be obtained as commercial products, or by the method described in the examples or a method analogous thereto.

Among compounds 37, those in which M ¹⁰ is NY ^14A can also be obtained by reacting R ⁴ -M ⁷ -OMs (compound 24) with Y ^14A NH ₂ .
Compounds 37 and 39 can be prepared in the same manner as compound 35.

Compound (IIf) was esterified / amidated in turn with Compound 24 and Compounds 37, 39 and 41 as described in Synthesis Route 8, and finally Compound 23 was allowed to act to introduce X ⁴ You can also get it.

(Wherein each group has the same meaning as above)

Compound (IIg) can be obtained from ethyl cyanoacetate according to synthetic route 9.

(In the formula, Et represents an ethyl group, LAH is lithium aluminum hydride, and other groups are as defined above)

Combine ethyl cyanoacetate with compound 24 in a solvent (eg, an aprotic solvent such as tetrahydrofuran), a base (eg, an inorganic base such as sodium hydride), and optionally an additive (eg, tetraiodide). Compound 42 can be obtained by reacting at an elevated temperature (for example, 60 ° C. or higher) in the presence of an additive such as butylammonium.

Compound 42 and Compound 26 in a solvent (eg, an aprotic solvent such as tetrahydrofuran), a base (eg, an inorganic base such as sodium hydride), and optionally an additive (eg, tetrabutyl iodide) 43 can be obtained by reacting at a high temperature (for example, 60 ° C. or more) in the presence of an additive such as ammonium.

Compound 43 in which R ⁴ and R ⁵ are the same can also be obtained by using an excess amount of compound 24 from ethyl cyanoacetate.

Compound 44 can be obtained by reducing compound 43 with an excess amount of lithium aluminum hydride (LAH) in a solvent (for example, an aprotic solvent such as tetrahydrofuran).

Compound 47 can be obtained by sequentially reacting compound 45, compound 46 and compound 23 with respect to compound 44 in the presence or absence of a solvent (eg, a halogen solvent such as chloroform). Compound 47 in which X ² , X ³ and X ⁴ are the same can also be obtained using 44 to an excess amount of compound 45.

Compound (IIg) can be obtained by reacting Compound 47 and Compound 21 by applying the same reaction conditions as in the esterification reaction of Compound 8 and Compound 9 in Synthesis Route 2. Incidentally, for example, by treating the (IIg) with a suitable anion exchange resins, it is also possible to convert the anion A ² of the compound (IIg) into another anion.

Compounds 45 and 46 are the same as compound 23.

Compound (IIh) can be obtained from dimethyl malonate according to synthesis route 10.

(In the formula, Me represents a methyl group, LAH is lithium aluminum hydride, and other groups are as defined above)

A dimethyl malonate and a compound 24 are mixed with a base (for example, an inorganic base such as cesium carbonate) and optionally an additive (for example, tetrabutyl iodide) in a solvent (for example, an aprotic solvent such as acetonitrile). Compound 48 can be obtained by reaction under heating (for example, 50 ° C.) in the presence of an additive such as ammonium.

Compound 50 is reacted with Compound 49 in the presence of acetic anhydride and a base (for example, an inorganic base such as sodium hydride) in a solvent (for example, an aprotic solvent such as acetonitrile). Obtainable.

Compound 51 can be obtained by reducing compound 50 with an excess amount of lithium aluminum hydride (LAH) in a solvent (for example, an aprotic solvent such as tetrahydrofuran).

Compound 52 is obtained by reacting Compound 51 with Compound 19 and Compound 21 under the same reaction conditions as in esterification of Compound 8 and Compound 9 in Synthesis Route 2.

Compound (IIh) is obtained by reacting compound 52 and compound 53 in the presence or absence of a solvent (eg, a halogen-based solvent such as chloroform) at room temperature or high temperature (eg, 100 ° C. or higher). . In addition, for example, by treating (IIh) with a suitable anion exchange resin, the anion A ² of the compound (IIh) can be converted into another anion.

Compound 49 is a commercially available product, or a method described in Examples or a method analogous thereto, or known from the literature (for example, “Helvetica Chimica Acta, Vol. 92, No. 8, 1644-1656”). Page, 2009 "etc.) or a method analogous thereto.

Compounds 54 and 56 can be obtained according to synthetic route 11.

(Wherein each group is as defined above)

Compound 54 can be obtained by protecting the hydroxy of compound 53.
Compound 53 can be obtained as a commercial product.

Compound 55 can be obtained by allowing compound 45 and compound 46 to act on compound 54.

Compound 56 can be obtained by deprotecting compound 55.

Compounds 58 to 65 can be obtained according to synthesis route 12.

(In the formula, Hal is a halogen atom such as chlorine, bromine, iodine, etc., and other groups are as defined above.)

Compound 58 can be obtained by protecting the hydroxy of compound 57.
Compound 57 can be obtained as a commercial product.

Compound 59 can be obtained by reacting Compound 58 with a halogenating reagent (for example, chlorine, bromine, iodine, iodine chloride, etc.).

Compound 60 is obtained by reacting compound 59 with ammonia. Compound 61 is obtained by allowing compound 45 to act on compound 60. Further, compound 63 is obtained by allowing compound 46 to act on compound 61.
Compound 63 can also be obtained by reacting compound 59 with compound 62.

Compound 63 is obtained by deprotecting compound 62.

Compound 64 is obtained by oxidizing compound 58 with an appropriate oxidizing agent (for example, potassium permanganate or Jones reagent).

Compounds 67 to 73 can be obtained according to synthesis route 13.

(Wherein each group is as defined above)

Compound 66 can be obtained by reacting compound 59 with cyanide (for example, sodium cyanide, potassium cyanide, lithium cyanide, etc.).

Compound 67 is obtained by reducing compound 66 with lithium aluminum hydride or the like.

Compound 68 is obtained by allowing Compound 45 to act on Compound 67. Compound 69 is obtained by allowing compound 46 to act on compound 68.

Compound 70 is obtained by deprotecting Compound 69.

Compound 71 is obtained by hydrolyzing compound 66 with a base (for example, sodium hydroxide or the like).

Compound 72 is obtained by reducing compound 71 by reduction (for example, borane or the like).

Compound 73 can be obtained by reacting compound 72 with a halogenating reagent (eg, chlorine, bromine, iodine, iodine chloride, etc.).

Compound 73, 68, 69, 71, 72, and 73 are subjected to the respective reactions from compound 59 in synthesis route 13 in order for compound 73, whereby each functional group (amino group, monoalkylamino group, dialkylamino group) is obtained. Groups, carboxylic acids, hydroxy and halogen) and quaternary carbons can be obtained in further extended compounds. By repeating this, the alkylene chain between each functional group and the quaternary carbon can be freely extended.

Compound 76 can be synthesized according to Synthesis Route 14.

(Wherein, M ¹³ is - (CH ₂₎ _p201 - a is, M ¹⁵ is - (CH ₂₎ _p202 - a is (wherein, p ²⁰¹ ~ p ²⁰² is an integer of 1 ~ 5), M ¹⁴ Is -O-, -CO-O- or -NY ^27A- , and PRO ⁴ is in accordance with M ¹⁴ , protecting group PRO ¹ for hydroxy, protecting group PRO ² for carboxylic acid, or amine Or a protective group PRO ³ (for example, a carbamate protecting group such as tert-butoxycarbonyl or benzyl)

Compound 74 can be obtained by the method described in Synthesis Routes 11 to 13 or a method analogous thereto.

Compound 75 can be obtained by appropriately protecting and deprotecting compound 74.

Compound 76 can be obtained from compound 75 as a raw material by the method described in Synthesis Routes 11 to 13 or a method analogous thereto.

Compounds 77 to 79 can be obtained by using compound 76 as a raw material, as described in Synthesis Route 15, and sequentially carrying out protection and deprotection, and a method according to Synthesis Route 14.

(Wherein, M ¹⁶ and M ¹⁷ are each - (CH ₂₎ _p203 - and _- (CH ₂₎ p204- a represents (wherein, p ²⁰³ ~ p ²⁰⁴ is an integer of 1-5), other Each group is as defined above)

Compounds 82, 84, 87, 89, 92 and 95 can be synthesized according to synthesis route 16.

(Wherein, M ¹⁸ is ^{^{_{- (CY 19 Y 20) p9}}} - or ^{^{_{- (CY 23 Y 24) p11}}} -Z 9 - (CY 25 Y 26) p12 -. Is also, b ¹ is

Ar is a p-nitrophenyl group, Hal is a halogen atom such as chlorine, bromine, iodine, etc., and other groups are as defined above. Note that when p13 is 0, N is the coupling carbon atom adjacent to the directly Z ^10. )

Compound 82 can be obtained by condensing compound 80 and compound 81 by esterification and then deprotecting.

Compound 84 can be obtained by condensing compound 83 and compound 81 by amidation and then deprotecting.

Compound 87 is obtained by condensing compound 85 and compound 86 by esterification and then deprotecting.

Compound 89 can be obtained by condensing compound 85 and compound 88 by amidation and then deprotecting.

Compound 92 is obtained by deprotecting Compound 90 and Compound 91 after a nucleophilic substitution reaction.

Compound 95 is obtained by deprotecting Compound 93 and Compound 94 after a transesterification reaction.

80, 83, 85, 90 and 93 can be obtained by the synthesis route 12-15 or a method according to these.

Among compounds 81, 86, 88, 91 and 94, those in which M ¹⁸ is — (CY ¹⁹ Y ²⁰ ) _p9 — are commercially available, or are the methods described in the examples or a method analogous thereto, or commercially available It can be obtained by converting the functional group of the product according to a conventional method.
In this case, b ¹ is

For compounds that are

(Wherein M ³⁷ is —OH, —CO ₂ H or NY ³⁸ (where Y ³⁸ is a hydrogen atom or an optionally substituted C1-C4 alkyl)), respectively. Appropriate fragments can be obtained by condensation by etherification, amination, esterification, amidation and the like.

Further, among the compounds 81,86,88,91 and 94, M ¹⁸ is ^{^{_{- (CY 23 Y 24) p11}}} -Z 9 - (CY 25 Y 26) p12 - a is one, the compounds 81,86,88, Of 91 and 94, M ¹⁸ is the same _as- (CY ¹⁹ Y ²⁰ ) _p9- , and the appropriate fragment corresponding to each is condensed by etherification, amination, esterification, amidation, etc. Can be obtained.

Hereinafter, the compound (III) can be obtained by the methods of Synthesis Routes 17 to 21 or methods according to these methods.

Compound (IIIa) can be obtained from Compound 96 according to Synthesis Route 17.

(Wherein each group is as defined above)

Compound (IIIa) is obtained by reacting Compound 96 and Compound 97 in the presence or absence of a solvent (eg, a halogen-based solvent such as chloroform) at room temperature or at a high temperature (eg, 100 ° C. or higher). . In addition, for example, by treating (IIIa) with an appropriate anion exchange resin, the anion A ³ of the compound (IIIa) can be converted to another anion.

Compound 96 can be obtained by the method described in Examples or a method analogous thereto, or the method described in the literature (US Patent Application Publication No. 2012/0172411) or a method analogous thereto.

The compound used for the reaction such as Compound 97 can be obtained as a commercially available product, by the method described in Examples or a method according thereto, or by a method known from the literature or a method according thereto.

Compound (IIIb) can be obtained from ethyl glyoxylate according to synthetic route 18.

(Wherein M ¹⁹ is ^{^{_{- (CY 50 Y 51) p23}}} - or ^{^{_{- (CY 54 Y 55) p25}}} -Z 17 - (CY 56 Y 57) p26 - ( wherein each group is respectively as defined above ) And b ²

And the other groups are as defined above. Note that when p33 is 0, N is the coupling carbon atom adjacent to the directly Z ^21. )

Compound 101 is obtained by sequentially reacting ethyl glyoxylate and Grignard reagents 98, 99 and 100 in a solvent (for example, an ether solvent such as tetrahydrofuran). The compound 101 having the same R ⁷ , R ⁸ and R ⁹ can also be obtained by reacting an excess amount of the compound 98 with ethyl glyoxylate.

Compound 101 and Compound 102 in a solvent (for example, a halogen-based solvent such as chloroform), a base (for example, an organic base such as triethylamine), a condensing agent (for example, 1-ethyl-3- (3-dimethylaminopropyl) Compound 103 is obtained by treatment with a condensing agent such as) carbodiimide hydrochloride) and an activator (an activator such as N, N-dimethylaminopyridine).

Compound (IIIb) is reacted with Compound 103 and Compound 97, 104 or 105 in the presence or absence of a solvent (for example, a halogen-based solvent such as chloroform) at room temperature or high temperature (for example, 100 ° C. or higher). ) Is obtained. Incidentally, for example, by treating the compound (IIIb) with a suitable anion exchange resins, it is also possible to convert the anion A ³ of the compound (IIIb) to another anion.

Compound 98 includes R ⁷ —OH (commercially available, obtained by the method described in Examples or a method analogous thereto), mesylation reagent (such as mesylic anhydride or mesylic chloride), bromide salt (magnesium bromide or Lithium bromide, etc.) and metal magnesium are obtained in this order. Compounds 99 and 100 are similar to compound 98.

Compounds 104 and 105 are the same as compound 23.
Compound 102 is the same as compound 54.

Compounds 108, 109, and 112 can be obtained from ammonia, ethyl formate, and compound 99, respectively, according to synthesis route 19.

(Wherein M ²¹ is —OH and M ²² is HO—CO— and M ²⁵ is —O—CO—, or M ²¹ is —NY ^45C —H and M ²² is HO—CO—. Yes M ²⁵ is -NY ^45C -CO-, M ²¹ is -CO-OH and M ²² is HO- and M ²⁵ is -CO-O-, or M ²¹ is -CO-OH And M ²² is H-NY ^14B -and M ²⁵ is -CO-NY ^14B- , and M ²⁰ is absent and M ²³ is-(CY ³⁹ Y ⁴⁰ ) _p18- or M ²⁰ M ²³ does not exist is ^{^{_{- (CY 41 Y 42) p19}}} -Z 14 - (CY 43 Y 44) p20 - or where, M ²⁰ is ^{^{^{-Z 13 - (CY 41 Y 42}}} ) p19 - a is M ²³ is ^{^{_{- (CY 43 Y 44) p20}}} - a further M ²⁴ is ^{^{-. (CY 63 Y 64)}} p29 -, or ^{^{_{- (CY 67 Y 68) p31}}} -Z 20 - (CY 69 Y 70) p32 - And the other groups are as defined above.)

Compound 108 is obtained by reacting ammonia, Compound 106, and Compound 107 under the same reaction conditions as those for synthesizing Compound 4 from ammonia in Synthesis Route 1.

Compound 109 is obtained by reacting ethyl formate, compound 99, and compound 100 under the same reaction conditions as those for synthesizing compound 101 from ethyl glyoxylate in synthesis route 18.

Compound 112 can be obtained by condensing compound 110 and compound 111 by esterification or amidation, followed by deprotection.

Compounds 106 and 107 are the same as compound 1.
The compound 110 in which M ²¹ is —OH is the same as R ¹ -L ¹ -OH described in the synthesis route 1. In addition, Compound 110 in which M ²¹ is —NY ^45C —H is the same as Compound 37. Further, Compound 110 in which M ²¹ is —CO—OH is the same as Compound 17.

Compound 111 is a commercially available product, a method described in Examples or a method similar thereto, or a method known from literature (for example, 4th edition, Experimental Chemistry Course 20, Synthesis of Organic Compounds II, 4th edition, p. .187, Maruzen (1992), etc.) or a method similar thereto.

Compounds (IIIc) and (IIId) can be obtained from compounds 108 and 112, and compounds 109 and 112, respectively, according to synthetic route 20.

(Wherein each group is as defined above)

Compound (IIIc) can be obtained by condensing compounds 108 and 112 by amidation and then allowing compound 114, 1115, or 116 to act.

Compound (IIId) can be obtained by condensing compounds 109 and 112 by esterification and then allowing compound 114, 1115, or 116 to act.

For example, the anion A ³ of the compound (IIIc) or (IIId) can be converted to another anion by treating the compound (IIIc) or (IIId) with a suitable anion exchange resin.

Each compound used in the reaction is as described above.

Hereinafter, the compound (IV) can be obtained by the methods of synthesis routes 11 to 12, or a method according to these methods.

Compound 127 can be obtained according to synthetic route 21.

(Wherein M ²⁶ and M ²⁷ are the same or different-(CY ⁹¹ Y ⁹² ) _p41- , and M ²⁸ and M ²⁹ are the same or different -O-CO- (CY ⁹¹ Y ⁹² ) _{p41 -or} -CO-O- (CY ⁹¹ Y ⁹² ) _p41- and M ³⁰ and M ³¹ are not the same or different, or -O-CO- (CY ⁹¹ Y ⁹² ) _p41 -or -CO-O- ( CY ⁹¹ Y ⁹² ) _p41- and the other groups are as defined above.

Compound 121 can be obtained by sequentially condensing compound 118, compound 119 and compound 120 by an esterification reaction, or by sequentially condensing compound 122, compound 123 and compound 124 by an esterification reaction.

Compound 125 is obtained by reacting Compound 121 with a deprotection reagent (for example, a deprotection reagent such as tetra-n-butylammonium fluoride) in a solvent (for example, an ether solvent such as tetrahydrofuran) or reacting with ethyl formate. In addition, it can be obtained by subjecting compound 126 and compound 127 to sequential addition reaction in a solvent (for example, an ether solvent such as tetrahydrofuran).

Compound 125 is mixed with an oxidizing agent (for example, an organic oxidizing agent such as Dess-Martin reagent or an inorganic oxidizing agent such as pyridinium chlorochromate) in a solvent (for example, an aprotic solvent such as chloroform). ) To give compound 128.

Compounds used for the reaction such as Compound 118, Compound 119, Compound 120, Compound 122, Compound 123, Compound 124, Compound 126 and Compound 127 are commercially available products, or the methods described in the Examples or a method analogous thereto, Alternatively, a method known from literature ("4th edition, Experimental Chemistry Course 22, Synthesis of Organic Compounds IV", 4th Edition, p.1, Maruzen (1992), "4th Edition, Experimental Chemistry Course 20, Synthesis of Organic Compounds II") , 4th edition, p.1, Maruzen (1992), and "4th edition experimental chemistry course 25: Synthesis of organic compounds VII", 4th edition, p.59, Maruzen (1991) etc.) or equivalent Can be obtained by the method.

Compound (IVa) can be obtained from Compound 128 according to Synthesis Route 22.

(Wherein each group is as defined above)

Compound 128 and Compound 129 may be combined with a reducing agent (for example, a hydride compound such as sodium borohydride or triacetoxyborohydride) in a solvent (for example, a halogen-based solvent such as 1,2-dichloroethane) or the like. Can be reacted in the presence of an additive (eg, an acid such as acetic acid) to give compound 130.

Compound 132 is obtained by reacting Compound 130 and Compound 131 in the presence of a base (for example, an inorganic base such as sodium hydroxide) at a high temperature (for example, 100 ° C. or higher). A solvent is not particularly required, but a solvent such as ethylene glycol may be used in some cases.

Compound (IVa) is obtained by reacting Compound 132 and Compound 133 in the presence or absence of a solvent (eg, a halogen-based solvent such as chloroform) at room temperature or high temperature (eg, 100 ° C. or higher). . For example, the anion A ⁴ of the compound (IVa) can be converted to another anion by treating (IVa) with an appropriate anion exchange resin.

Compounds used for the reaction such as Compound 129, Compound 131, and Compound 133 are commercially available, or the methods described in Examples or a method analogous thereto, or methods known from the literature (for example, International Publication No. 2010/042877 No., International Publication No. 2010/054401, “Fifth Edition Experimental Chemistry Lecture 13, Synthesis of Organic Compounds I”, Fifth Edition, p.374, Maruzen (2005), etc.) It can be obtained.

Compound (IVb) can be obtained according to synthesis route 23.

(In the formula, M ³² does not exist or is —Z ²⁷ — (CY ⁹³ Y ⁹⁴ ) _p42 —, Ns represents an o-nitrobenzenesulfonyl group, and other groups are as defined above)

After reacting compound 134, compound 135, triphenylphosphine, and diethyl azodicarboxylate, thiol (eg, dodecane-1-thiol, thiophenol, etc.) is allowed to act on the resulting condensate to remove Ns. Gives compound 136.

Compound 136 is obtained by amidating compounds 136 and 137.

Compound (IVb) can be obtained by reacting Compound 138 with 139. For example, the anion A ⁴ of the compound (IVb) can be converted to another anion by treating the compound (IVb) with an appropriate anion exchange resin.

Compound 134 is obtained by allowing o-nitrobenzenesulfonyl chloride to act on R ¹¹ -L ¹⁴ -NH ₂ . R ¹¹ -L ¹⁴ -NH ₂ is a commercially available product, a method described in Examples or a method analogous thereto, or a method known from literature (for example, “4th edition Experimental Chemistry Course 20 Synthesis of Organic Compounds II , 4th edition, p.279, Maruzen (1992), etc.) or a method similar thereto.

Compounds used for the reaction such as compounds 135, 137 and 139 can be obtained by any of the methods described above.

Compound (V′a) can be obtained according to synthesis route 24

(Wherein, DMTr is 2 ', 2''- represents a dimethoxytrityl group, M ³³ ^{^{is, - (CY 123 R 124)}} p54 -, - (CY 125 Y 126) p55 -Z 35 - (CY 127 R 128 ) _p56 - or ^{^{_{- (CY 129 R 130) p57}}} -Z 36 - (CY 131 Y 132) p58 -Z 37 - (CY 133 Y 134) p59 - a Awarashi, M ^34, M ^35, and M ³⁶ are each Independently -O- or -CO-O-, and other groups are as defined above)

Compound 141 can be obtained by reacting Compound 140 with 2 ′, 2 ″ -dimethoxytrityl chloride.

Compound 142 is obtained by subjecting Compound 141 to three-stage etherification or esterification.

Compound 143 is obtained by treating compound 142 with an acid.

Compound 144 is obtained by activating compound 143 with a halogenating reagent and then treating with the corresponding amine compound.

Compound (V′a) can be obtained by reacting compound 145 with compound 144. Incidentally, for example, by treating the (V'a) in a suitable anion exchange resins, it is also possible to convert the anion A ⁵ of the compound (V'a) in a different anion.

Compound 140 is a commercially available product, a natural product, or a method described in Examples or a method analogous thereto, or a method known in the literature (for example, “(The Organic Chemistry of Sugar (The Organic Chemistry Chemistry of Sugars), Daniel E. 他 Levy, et al., the method described in Tyler & Francis, 2005, etc.) or a method similar thereto.

Compound (V ″ a) can be obtained by the same method as in synthesis route 25 using compound 146 as a raw material.

(Wherein each group is as defined above)

Compound 146 can be obtained as a commercially available product, as a natural product, or by a method described in the Examples or a method analogous thereto, or a method known from the literature (for example, “(The Organic Chemistry of Sugar (The Organic Chemistry Chemistry of Sugars), Daniel E. 他 Levy, et al., the method described in Tyler & Francis, 2005, etc.) or a method similar thereto.

Compounds (I) to (V ″) can be obtained by appropriately combining any one of the above synthesis routes 1 to 25 or a method according to these methods.

The lipid represented by the compound (CL-I) can be obtained by the method described in International Publication No. 2013/089151, or a method analogous thereto.

The lipid represented by the compound (CL-II) can be obtained by the method described in International Publication No. 2011/136368 or a method analogous thereto.

Lipids represented by compound (CL-III), compound (CL-IV) and compound (CL-V) can be obtained by the method described in International Publication No. 2014/007398 or a method analogous thereto.

The lipid represented by the compound (CL-VI) can be obtained by the method described in International Publication No. 2010/042877 or a method analogous thereto.

Compound (CL-VII) can be obtained by the method described in International Publication No. 2010/054401, the method described in International Publication No. 2013/059496, or a method analogous thereto.

Compound (CL-VIII) can be obtained by the method described in International Publication No. 2016/002753 or a method analogous thereto.

Compound (CL-IX) can be obtained by the method described below or a method analogous thereto.

The production method of the compound of the present invention will be described. In addition, in the production method shown below, when a defined group changes under the conditions of the production method or is inappropriate for carrying out the production method, introduction and removal of a protective group commonly used in organic synthetic chemistry Methods [e.g., Methods described in Protective Groups in Organic Synthesis, third edition, by TWGreene, John Wiley & Sons Inc. (1999)], etc. Can be used to produce the target compound. Further, the order of reaction steps such as introduction of substituents can be changed as necessary.

Production method 1
Among compounds (CL-IX), compound (CL-IXa) in which X ¹¹⁵ and X ¹¹⁶ are both hydrogen atoms and compound (CL-IXb) in which X ¹¹⁵ and X ¹¹⁶ are the same are produced by the following method. be able to.

(Wherein R ¹¹⁸ , R ¹¹⁹ , M ¹⁰¹ , M ¹⁰² , L ¹¹⁸ and L ¹¹⁹ have the same meanings as defined above, and X in IX-IIIa and IX-IIIb is the same or different, and represents a chlorine atom, bromine atom, Represents a leaving group such as iodine atom, trifluoromethanesulfonyloxy, methanesulfonyloxy, benzenesulfonyloxy, p-toluenesulfonyloxy, R ¹³⁷ is a hydrogen atom, methyl or ethyl, and R ¹³⁸ is a hydrogen atom or methyl Or R ¹³⁷ and R ¹³⁸ together with adjacent carbons form a cyclopropyl ring (provided that when R ¹³⁷ is a hydrogen atom or ethyl, R ¹³⁸ is not methyl))

Step 26 and Step 27
Compound (IX-IIa) is obtained by reacting 2-amino-2-methyl-1,3-propanediol and compound (IX-IIIa) at room temperature in the presence of 1 to 10 equivalents of a base without solvent or in a solvent. It can be produced by reacting at a temperature between 200 ° C. for 5 minutes to 100 hours. Further, compound (CL-IXa) is obtained by reacting compound (IX-IIa) and compound (IX-IIIb) in the presence of 1 to 10 equivalents of base without solvent or in a solvent at a temperature between room temperature and 200 ° C. Thus, it can be produced by reacting for 5 minutes to 100 hours.

Examples of the solvent include dichloromethane, 1,2-dichloroethane, toluene, diethyl ether, tetrahydrofuran, 1,2-dimethoxyethane, 1,4-dioxane, pyridine and the like, and these may be used alone or in combination. it can.

Examples of the base include sodium methoxide, potassium tert-butoxide, sodium hydride, lithium diisopropylamide, hexamethyldisilazane lithium, hexamethyldisilazane sodium, n-butyllithium and the like.

Compound (IX-IIIa) and Compound (IX-IIIb) are commercially available products or known methods (for example, “5th edition Experimental Chemistry Course 13 Synthesis of Organic Compounds I”, 5th edition, p.374, Maruzen ( 2005)) or a method based thereon.

Compound (CL-IXa) when R ¹¹⁸ -M ¹⁰¹ -L ¹¹⁸ and R ¹¹⁹ -M ¹⁰² -L ¹¹⁹ are the same can be obtained by using 2 equivalents or more of compound (IX-IIIa) in Step 26 Can do.

2-Amino-2-methyl-1,3-propanediol can be obtained as a commercial product.

Step 28
Compound (CL-IXb) is obtained by mixing compound (CL-IXa) with 2 to 20 equivalents of compound (IX-IV), preferably 1 equivalent to a large excess of reducing agent and preferably 1 to 10 equivalents in a solvent. It can be produced by reacting at a temperature between −20 ° C. and 150 ° C. for 5 minutes to 72 hours in the presence of an equivalent amount of acid.

Examples of the solvent include methanol, ethanol, tert-butyl alcohol, dichloromethane, chloroform, 1,2-dichloroethane, toluene, ethyl acetate, acetonitrile, diethyl ether, tetrahydrofuran, 1,2-dimethoxyethane, 1,4-dioxane, N , N-dimethylformamide, N, N-dimethylacetamide, N-methylpyrrolidone, water and the like, and these may be used alone or in combination.

Examples of the reducing agent include sodium triacetoxyborohydride and sodium cyanoborohydride.

Examples of the acid include hydrochloric acid and acetic acid.

Compound (IX-IV) can be obtained as a commercial product.

Production method 2
Of the compounds (CL-IX), compound X ¹¹⁵ and X ¹¹⁶ are different (CL-IXc) and (CL-IXd) can be prepared by the following method.

(Wherein R ¹¹⁸ , R ¹¹⁹ , M ¹⁰¹ , M ¹⁰² , L ¹¹⁸ , L ¹¹⁹ , R ¹³⁷ , R ¹³⁸ and X are as defined above, R ¹³⁹ is as defined in X ¹¹⁵ , and PG is protected. Represents a group.)

Step 29
Compound (IX-IIb) is compound (CL-IXa) that is commonly used in synthetic organic chemistry (e.g., Protective Groups in Organic Synthesis, third edition, Green (TWGreene ) By John Wiley & Sons Inc. (1999), etc.].

Process 30
Compound (IX-IIc) is obtained by reacting Compound (IX-IIb) and Compound (IX-IIIc) in the presence of 1 to 10 equivalents of a base without solvent or in a solvent at a temperature between −20 ° C. and 150 ° C. Thus, it can be produced by reacting for 5 minutes to 72 hours.

Examples of the solvent include dichloromethane, 1,2-dichloroethane, toluene, diethyl ether, tetrahydrofuran, 1,2-dimethoxyethane, 1,4-dioxane, pyridine, N, N-dimethylformamide, N, N-dimethylacetamide and the like. These may be used alone or in admixture.

Examples of the base include sodium methoxide, potassium tert-butoxide, sodium hydride, lithium diisopropylamide, hexamethyldisilazane lithium, hexamethyldisilazane sodium, n-butyllithium, potassium carbonate, cesium carbonate, triethylamine and the like. .

Compound (IX-IIIc) can be obtained as a commercial product.

Step 31
Compound (CL-IXc) can be obtained by removing protecting group PG of compound (IX-IIc) by an appropriate method. Methods for removing protecting groups include those commonly used in organic synthetic chemistry (for example, Protective Groups in Organic Synthesis, third edition, TW Greene, John The removal method described in Wiley & Sons Inc. (1999), etc.] can be used, whereby the target compound can be produced.

Process 32
Compound (CL-IXd) is obtained by combining compound (CL-IXc) with 1 to 10 equivalents of compound (IX-IV), preferably 1 equivalent to a large excess of a reducing agent and preferably 1 to 10 equivalents in a solvent. It can be produced by reacting at a temperature between −20 ° C. and 150 ° C. for 5 minutes to 72 hours in the presence of an equivalent amount of acid.

Examples of the solvent, reducing agent, and acid include those exemplified in Step 28.

Production method 3
Among the compounds (CL-IX), compounds (CL-IXc ′) and (CL-IXd ′) in which M ¹⁰¹ and M ¹⁰² are —OC (O) — can also be produced by the following method.

Wherein R ¹¹⁸ , R ¹¹⁹ , M ¹⁰¹ , M ¹⁰² , L ¹¹⁸ , L ¹¹⁹ , R ¹³⁷ , R ¹³⁸ , R ¹³⁹ and PG are as defined above, and B and B ^′ are linear or branched C1-C16 alkyl or C2-C16 alkenyl in the form of

Step 33
Compound (IX-IId) can be produced by reacting compound (IX-IIc ′) and an oxidizing agent in a solvent at a temperature between −20 ° C. and 150 ° C. for 5 minutes to 72 hours.

Examples of the oxidizing agent include ozone, osmium tetroxide / sodium periodate, osmium tetroxide / lead tetraacetate, and the like.

Examples of the solvent include those exemplified in Step 28.

Compound (IX-IIc ′) can be produced by the method described in Production Method 2.

Step 34
Compound (IX-IIe) can be produced by reacting compound (IX-IId) and an oxidizing agent in a solvent at a temperature between −20 ° C. and 150 ° C. for 5 minutes to 72 hours.

Examples of the oxidizing agent include Jones reagent, pyridinium dichromate, ruthenium tetroxide, sodium chlorite and the like.

Solvents include tert-butyl alcohol, dichloromethane, chloroform, 1,2-dichloroethane, toluene, ethyl acetate, acetone, acetonitrile, diethyl ether, tetrahydrofuran, 1,2-dimethoxyethane, 1,4-dioxane, N, N- Examples thereof include dimethylformamide, N, N-dimethylacetamide, N-methylpyrrolidone, water and the like, and these can be used alone or in combination.

Step 35 and Step 36
Compound (IX-IIf) is obtained by reacting Compound (IX-IIe) and Compound (IX-Va) at room temperature and in the presence of 1 to 10 equivalents of a condensing agent and 1 to 10 equivalents of a base without solvent or in a solvent. It can be produced by reacting at a temperature between 0 ° C. and 5 minutes to 100 hours. Further, Compound (IX-IIc ″) is obtained by combining Compound (IX-IIf) and Compound (IX-Vb) in the absence of solvent or in a solvent in the presence of 1 to 10 equivalents of condensing agent and 1 to 10 equivalents of base. The reaction can be carried out at a temperature between room temperature and 200 ° C. for 5 minutes to 100 hours.

Examples of the solvent include dichloromethane, chloroform, 1,2-dichloroethane, toluene, ethyl acetate, acetonitrile, diethyl ether, tetrahydrofuran, 1,2-dimethoxyethane, dioxane, N, N-dimethylformamide, N, N-dimethylacetamide, N-methylpyrrolidone, pyridine and the like can be mentioned, and these can be used alone or in combination.

Examples of the condensing agent include 1-ethyl-3- (3-dimethylaminopropyl) carbodiimide hydrochloride, N, N′-dicyclohexylcarbodiimide, 4- (4,6-dimethoxy-1,3,5-triazine-2- ジン yl). ) -4-Methylmorpholinium chloride n hydrate, 1H-benzotriazol-1-yloxytris (dimethylamino) phosphonium hexafluorophosphate, O- (7-azabenzotriazol-1-yl) -N , N, N ′, N ′,-tetramethyluronium hexafluorophosphate and the like.

Examples of the base include potassium carbonate, cesium carbonate, triethylamine, N, N-diisopropylethylamine, N-methylmorpholine, pyridine and the like.

Compound (IX-Va) and compound (IX-Vb) can be obtained as commercial products.

The compound (IX-IIc ″) in the case where R ¹¹⁸ and R ¹¹⁹ are the same can be obtained by using 2 equivalents or more of the compound (IX-Va) in Step 35.

Step 37
Compound (CL-IXc ′) is obtained by removing the protecting group PG of compound (IX-IIc ″) by an appropriate method. Methods for removing protecting groups include those commonly used in organic synthetic chemistry (for example, Protective Groups in Organic Synthesis, third edition, TW Greene, John The removal method described in Wiley & Sons Inc. (1999), etc.] can be used, whereby the target compound can be produced.

Step 38
Compound (CL-IXd ′) is obtained by combining compound (CL-IXc ′) with 1 to 10 equivalents of compound (IX-IV), preferably 1 equivalent to a large excess of reducing agent in a solvent and preferably 1 It can be produced by reacting at a temperature between −20 ° C. and 150 ° C. for 5 minutes to 72 hours in the presence of ˜10 equivalents of acid.

Examples of the solvent and the acid include those exemplified in Step 28.

Of the compounds (CL-IX), compounds other than the compounds (CL-IXa) to (CL-IXd) can be obtained by adopting materials and reagents suitable for the structure of the target compound. It can be produced according to the production method or by applying a general production method commonly used in organic synthetic chemistry.

The intermediates and target compounds in each of the above production methods should be isolated and purified by subjecting them to separation and purification methods commonly used in organic synthetic chemistry, such as filtration, extraction, washing, drying, concentration, and various recrystallization chromatography. Can do. The intermediate can be subjected to the next reaction without any particular purification.

R ¹¹⁵ and R ¹¹⁶ are the same or different and are a hydrogen atom or C1-C3 alkyl.
R ¹¹⁵ and R ¹¹⁶ are the same or different and are preferably a hydrogen atom, methyl, ethyl, or propyl, more preferably a hydrogen atom or methyl.
The combination of (R ¹¹⁵ , R ¹¹⁶ ) is preferably (hydrogen atom, hydrogen atom), (hydrogen atom, methyl), (methyl, methyl), more preferably (hydrogen atom, methyl), (methyl, Methyl).

L ¹¹⁸ and L ¹¹⁹ are the same or different and are linear or branched C8-C24 alkylene or C8-C24 alkenylene.
When L ¹¹⁸ and L ¹¹⁹ are the same or different and are alkylene, they are preferably linear C8-C24 alkylene, more preferably linear C8-C20 alkylene, and even more preferably linear C8-C12 alkylene.
L ¹¹⁸ and L ¹¹⁹ are the same or different and are preferably octylene, nonylene, undecylene, tridecylene, pentadecylene, and more preferably octylene, nonylene, undecylene.
When L ¹¹⁸ and L ¹¹⁹ are the same or different and are alkenylene, they are preferably linear C8-C24 alkenylene, more preferably linear C10-C20 alkenylene, and even more preferably linear C10-C12 alkenylene.
L ¹¹⁸ and L ¹¹⁹ are the same or different, preferably (Z) -undec-9-enylene, (Z) -tridec-11-enylene, (Z) -tetradec-9-enylene, (Z) -hexadeca-9 -Enylene, (Z) -octadeca-9-enylene, (Z) -octadeca-11-enylene, (9Z, 12Z) -octadeca-9,12-dienylene.
L ¹¹⁸ and L ¹¹⁹ are preferably the same.

M ¹⁰¹ and M ¹⁰² are the same or different, and -C = C-, -OC (O)-, -C (O) O-, -SC (O)-, -C (O) S-, -OC ( S)-, -C (S) O-, -SS-, -C (R ^'' ) = N-, -N = C (R ^'' )-, -C (R ^'' ) = NO-,- ON = C (R ^'' )-, -N (R ^'' ) C (O)-, -C (O) N (R ^'' )-, -N (R ^'' ) C (S)-,- ^{C (S) N (R '} ') -, - N (R '') C (O) N (R ''') -, - N (R 3) C (O) O -, - OC (O) N (R ^'' )-and -OC (O) O-.
M ¹⁰¹ and M ¹⁰² are the same or different and are preferably -C = C-, -OC (O)-, -C (O) O-, -C (O) (NR ^'' )-, -N (R ^'' ) C (O)-, -N (R ^'' ) C (O)-, -N (R ^'' ) C (O) N (R ^''' )-, -N (R ^'' ) C (O) O-, -OC (O) N (R ^'' )-, -OC (O) O-, more preferably -C = C-, -OC (O)-, -C (O) O-.
The bond of each structure of M ¹⁰¹ and M ¹⁰² will be described by taking —OC (O) — as an example, which means that the structure is R ¹¹⁸ —OC (O) —L ¹¹⁸ .
M ¹⁰¹ and M ¹⁰² are preferably the same.

R ^'in M ¹⁰¹ and M ^{^102'} and R ^'''the same or different, is a hydrogen atom or a C1-C3 alkyl.
R ^″ and R ^{′ ″} are preferably a hydrogen atom, methyl, ethyl or propyl, more preferably a hydrogen atom or methyl, and even more preferably a hydrogen atom.

R ¹¹⁸ and R ¹¹⁹ are the same or different and are linear or branched C1-C16 alkyl or C2-C16 alkenyl.
When R ¹¹⁸ and R ¹¹⁹ are the same or different and are alkyl, they are preferably linear C1-C16 alkyl, more preferably linear C2-C9 alkyl.
R ¹¹⁸ and R ¹¹⁹ are the same or different and are preferably pentyl, octyl, nonyl, decyl, dodecyl.
When R ¹¹⁸ and R ¹¹⁹ are the same or different and are alkenyl, they are preferably linear C 2 -C 16 alkenyl, more preferably linear C 3 -C 9 alkenyl.
R ¹¹⁸ and R ¹¹⁹ are the same or different, preferably (Z) -hept-2-ene, (Z) -oct-2-ene, (Z) -non-2-ene, (Z) -nona-3 -Ene, nona-8-ene, (Z) -dodec-2-ene, (Z) -tridec-2-ene.
R ¹¹⁸ and R ¹¹⁹ are preferably the same.

R ¹¹⁸ -M ¹⁰¹ -L ¹¹⁸ and R ¹¹⁹ -M ¹⁰² -L ¹¹⁹ are the same or different, and R ¹¹⁸ and R ¹¹⁹ , M ¹⁰¹ and M ¹⁰² , and L ¹¹⁸ and L ¹¹⁹ are from the structure described for each group. It may be a combination of
R ¹¹⁸ -M ¹⁰¹ -L ¹¹⁸ and R ¹¹⁹ -M ¹⁰² -L ¹¹⁹ are preferably the same.
R ¹¹⁸ -M ¹⁰¹ -L ¹¹⁸ and R ¹¹⁹ -M ¹⁰² -L ¹¹⁹ are the same or different, preferably (Z) -tetradec-9-enyl, (Z) -hexadeca-9-enyl, (Z) -octadeca -9-enyl, (E) -octadeca-9-enyl, (Z) -octadeca-11-enyl, (9Z, 12Z) -octadeca-9,12-dienyl, (9Z, 12Z, 15Z) -octadeca-9 , 12,15-trienyl, (Z) -icosa-11-enyl, (11Z, 14Z) -icosa-11,14-dienyl and (Z) -docosa-13-enyl, more preferably The group consisting of (Z) -hexadec-9-enyl, (Z) -octadeca-9-enyl, (9Z, 12Z) -octadeca-9,12-dienyl and (11Z, 14Z) -icosa-11,14-dienyl Chosen from.
R ¹¹⁸ -M ¹⁰¹ -L ¹¹⁸ and R ¹¹⁹ -M ¹⁰² -L ¹¹⁹ are the same or different and are preferably the following structures (1) to (5), more preferably the following structures (1) to (5): (5).

[Where n is an integer of 1-4]

The lipid represented by the compound (CL-X) can be obtained by the method described in International Publication No. 2009/129385 or a method analogous thereto.

The lipid represented by the compound (CL-XI) can be obtained by the method described in International Publication No. 2013/1491401, or a method analogous thereto.

The lipid represented by the compound (CL-XII) can be obtained by the method described in International Publication No. 2009/129395 or a method analogous thereto.

The lipid represented by the compound (CL-XIII) can be obtained by the method described in International Publication No. 2013/059496 or a method analogous thereto.

The lipid represented by the compound (CL-XIV) can be obtained by the method described in International Publication No. 2011/149733 or a method analogous thereto.

The lipid represented by the formula (CL-XV) can be obtained by the method described in International Publication No. 2011/153493 or a method analogous thereto.

The lipid represented by the formula (CL-XVI) can be obtained by the method described in International Publication No. 2015/074085 or a method analogous thereto.

The lipid represented by the formula (CL-XVII) can be obtained by the method described in International Publication No. 2012/170952 or a method analogous thereto.

Specific examples of lipid A in the present invention are shown in Tables 11 to 26, but lipid A is not limited thereto.

Lipid A in the nucleic acid-containing lipid nanoparticles of the present invention is represented by the formula (I), formula (II), formula (III), formula (IV), formula (V ′), and formula (V ″). Among these, lipids represented by formula (II), formula (V ′), and formula (V ″) are preferable, and lipids represented by formula (II) and formula (V ′) are more preferable.
Among the lipids represented by the formula (II), preferably, at least one of R ⁴ to R ^{6 in} the formula (II) is a linear C8-C24 alkyl lipid, more preferably, Two of R ⁴ to R ⁶ in the formula (II) are lipids that are linear C8-C24 alkyl, and more preferably, R ⁴ to R ⁶ in the formula (II) are all linear C8 It is a lipid that is -C24 alkyl.
The lipid B to be combined with a lipid selected from the group consisting of the formula (II), the formula (V ′), and the formula (V ″) is preferably the formula (CL-I), the formula (CL-II), the formula (CL-III), Formula (CL-IV), Formula (CL-V), Formula (CL-VI), Formula (CL-VII), Formula (CL-VIII), Formula (CL-IX), Formula ( CL-XII) and a lipid represented by the formula (CL-XIV).
Among the lipids represented by the formula (CL-II), preferably, L ¹⁰⁶ and L ¹⁰⁷ in the formula (CL-II) are combined to form a single bond or C2-C8 alkylene, and p ¹⁰¹ and p ¹⁰² are lipids that are integers of 1 to 3, more preferably, L ¹⁰⁶ and L ¹⁰⁷ in formula (CL-II) together form a single bond, and p ¹⁰¹ and p ¹⁰² Is a lipid that is 1.

The nucleic acid used in the present invention may be any molecule as long as it is a molecule obtained by polymerizing nucleotides and / or molecules having functions equivalent to nucleotides, for example, ribonucleic acid that is a polymer of ribonucleotides. (RNA), deoxyribonucleic acid (DNA) which is a polymer of deoxyribonucleotides, chimeric nucleic acids composed of RNA and DNA, and nucleotides in which at least one nucleotide of these nucleic acids is replaced with a molecule having a function equivalent to that nucleotide A polymer etc. are mentioned. In addition, the nucleic acid of the present invention also includes a derivative containing at least a part of the structure of a molecule obtained by polymerizing nucleotides and / or molecules having functions equivalent to nucleotides. In the present invention, uracil U and thymine T can be replaced with each other.

Examples of molecules having a function equivalent to nucleotides include nucleotide derivatives.

The nucleotide derivative may be any molecule as long as it is a modified nucleotide, for example, it improves nuclease resistance or stabilizes from other degradation factors compared to RNA or DNA. In order to increase the affinity with the complementary strand nucleic acid, to increase cell permeability, or to visualize the molecule, a molecule in which ribonucleotides or deoxyribonucleotides are modified is preferably used.

Examples of nucleotide derivatives include sugar moiety-modified nucleotides, phosphodiester bond-modified nucleotides, base-modified nucleotides, and the like.

The sugar moiety-modified nucleotide may be any nucleotide as long as it is modified or substituted with an arbitrary substituent on a part or all of the sugar sugar chemical structure, or substituted with an arbitrary atom. 2'-modified nucleotides are preferably used.

Examples of the modifying group in the sugar moiety-modified nucleotide include 2′-cyano, 2′-alkyl, 2′-substituted alkyl, 2′-alkenyl, 2′-substituted alkenyl, 2′-halogen and 2′-O-cyano. 2'-O-alkyl, 2'-O-substituted alkyl, 2'-O-alkenyl, 2'-O-substituted alkenyl, 2'-S-alkyl, 2'-S-substituted alkyl, 2'-S -Alkenyl, 2'-S-substituted alkenyl, 2'-amino, 2'-NH-alkyl, 2'-NH-substituted alkyl, 2'-NH-alkenyl, 2'-NH-substituted alkenyl, 2'-SO -Alkyl, 2'-SO-substituted alkyl, 2'-carboxy, 2'-CO-alkyl, 2'-CO-substituted alkyl, 2'-Se-alkyl, 2'-Se-substituted alkyl, 2'-SiH ₂ -alkyl, 2'-SiH ₂ -substituted alkyl, 2'-ONO ₂ , 2'-NO ₂ , 2'-N ₃ , 2'-amino acid residues (hydroxys removed from the carboxylic acid of the amino acid) 2'-O-amino acid residues (synonymous with the above amino acid residues) and the like.

Examples of sugar-modified nucleotides include, for example, a crosslinked structure-type artificial nucleic acid (BNA) having a structure in which a modification group at the 2 ′ position is crosslinked to a carbon atom at the 4 ′ position, more specifically, the 2 ′ position. Locked Nucleic Acid (LNA) in which the oxygen atom and 4'-position carbon atom are bridged via methylene, and Ethylene bridged nucleic acid (ENA) [Nucleic Acid Research, 32, e175 (2004)] and the like, and these are included in 2′-modified nucleotides.

Peptide nucleic acid (PNA) [Acc. Chem. Res., 32, 624 (1999)], oxypeptide nucleic acid (OPNA) [J. Am. Chem. Soc., 123, 4653 (2001)] And peptide ribonucleic acid (PRNA) [J. Am. Chem. Soc., 122, 6900 (2000)].

2'-cyano, 2'-halogen, 2'-O-cyano, 2'-alkyl, 2'-substituted alkyl, 2'-O-alkyl, 2'-O-substituted as a modifying group in sugar modified nucleotide Preferred are alkyl, 2′-O-alkenyl, 2′-O-substituted alkenyl, 2′-Se-alkyl or 2′-Se-substituted alkyl, 2′-cyano, 2′-fluoro, 2′-chloro, 2'-bromo, 2'-trifluoromethyl, 2'-O-methyl, 2'-O-ethyl, 2'-O-isopropyl, 2'-O-trifluoromethyl, 2'-O- [2- (Methoxy) ethyl], 2'-O- (3-aminopropyl), 2'-O- [2- (N, N-dimethylaminooxy) ethyl], 2'-O- [3- (N, N -Dimethylamino) propyl], 2'-O- {2- [2- (N, N-dimethylamino) ethoxy] ethyl}, 2'-O- [2- (methylamino) -2-oxoethyl] or 2 '-Se-methyl and the like are more preferable, 2'-O-methyl, 2'-O-ethyl, 2'-fluoro and the like are more preferable, and 2'-O-methyl Or 2'-O-ethyl is most preferred.

The preferred range of the modifying group in the sugar moiety-modified nucleotide can also be defined from its size, preferably from the size of fluoro to the size of -O-butyl, and from the size of -O-methyl- Those corresponding to the size of O-ethyl are more preferred.

Examples of the alkyl in the modifying group in the sugar-modified nucleotide include C1-C6 alkyl, and more specifically, methyl, ethyl, propyl, isopropyl, butyl, isobutyl, sec-butyl, tert-butyl, pentyl, isopentyl. And C1-C6 alkyl such as neopentyl or hexyl.

Examples of the alkenyl in the modifying group in the sugar moiety-modified nucleotide include C3-C6 alkenyl, and more specifically, C3-C6 alkenyl such as allyl, 1-propenyl, butenyl, pentenyl, hexenyl and the like.

Examples of the halogen in the modifying group in the sugar moiety-modified nucleotide include a fluorine atom, a chlorine atom, a bromine atom or an iodine atom.

Examples of amino acids in amino acid residues include aliphatic amino acids (specifically, glycine, alanine, valine, leucine, isoleucine, etc.), hydroxy amino acids (specifically, serine, threonine, etc.), acidic amino acids (specifically, Aspartic acid, glutamic acid, etc.), acidic amino acid amides (specifically, asparagine, glutamine, etc.), basic amino acids (specifically, lysine, hydroxylysine, arginine, ornithine, etc.), sulfur-containing amino acids (specifically, Specifically, cysteine, cystine, methionine and the like) or imino acid (specifically, proline, 4-hydroxyproline and the like) and the like.

Examples of the substituted alkyl in the modified group in the sugar-modified nucleotide and the substituent in the substituted alkenyl include halogen (as defined above), hydroxy, sulfanyl, amino, oxo, -O-alkyl (the alkyl portion of the -O-alkyl is The same as C1-C6 alkyl in the modifying group), -S-alkyl (the alkyl part of the -S-alkyl is synonymous with C1-C6 alkyl in the modifying group), -NH-alkyl (the alkyl of -NH-alkyl) Part is synonymous with C1-C6 alkyl in the modifying group), dialkylaminooxy (the two alkyl parts of the dialkylaminooxy are the same or different and are synonymous with C1-C6 alkyl in the modifying group), dialkylamino (the dialkylamino The two alkyl moieties are the same or different and have the same meaning as C1-C6 alkyl in the above-mentioned modifying group) or dialkylaminoalkyl Xi (the two alkyl moieties of the dialkylaminoalkyloxy are the same or different and have the same meaning as the C1-C6 alkyl in the modifying group, and the alkylene moiety has one hydrogen atom removed from the C1-C6 alkyl in the modifying group. The number of substitutions is preferably 1 to 3.

The phosphodiester bond-modified nucleotide is any nucleotide that has been modified or substituted with an arbitrary substituent for a part or all of the chemical structure of the phosphodiester bond of the nucleotide, or with any atom. For example, a nucleotide in which a phosphodiester bond is replaced with a phosphorothioate bond, a nucleotide in which a phosphodiester bond is replaced with a phosphorodithioate bond, a nucleotide in which a phosphodiester bond is replaced with an alkylphosphonate bond, or a phosphate Examples include nucleotides in which a diester bond is substituted with a phosphoramidate bond.

As the base-modified nucleotide, any or all of the nucleotide base chemical structure modified or substituted with an arbitrary substituent or substituted with an arbitrary atom may be used. In which oxygen atom is substituted with sulfur atom, hydrogen atom is substituted with C1-C6 alkyl group, methyl group is substituted with hydrogen atom or C2-C6 alkyl group, amino group is C1-C6 Examples include those protected with a protecting group such as an alkyl group and a C1-C6 alkanoyl group.

Further, as nucleotide derivatives, nucleotides or sugar moieties, nucleotide derivatives modified with at least one of phosphodiester bonds or bases, lipids, phospholipids, phenazine, folate, phenanthridine, anthraquinone, acridine, fluorescein, rhodamine, coumarin and Examples include dyes and other chemical substances added. Specifically, 5'-polyamine-added nucleotide derivatives, cholesterol-added nucleotide derivatives, steroid-added nucleotide derivatives, bile acid-added nucleotide derivatives, vitamin-added nucleotide derivatives, green Examples include fluorescent dye (Cy3) addition nucleotide derivatives, red fluorescent dye (Cy5) addition nucleotide derivatives, fluorescein (6-FAM) addition nucleotide derivatives, and biotin addition nucleotide derivatives. The

In the nucleic acid used in the present invention, a nucleotide or nucleotide derivative is a structure obtained by combining at least one of these with other nucleotides or nucleotide derivatives in the nucleic acid, an alkylene structure, a peptide structure, a nucleotide structure, an ether structure, an ester structure, or the like. A cross-linked structure such as

The nucleic acid used in the present invention preferably has a molecular weight of 1,000 kDa or less, more preferably 100 kDa or less, and even more preferably 30 kDa or less. The nucleic acid used in the present invention preferably includes a nucleic acid that suppresses the expression of the target gene, and more preferably includes a nucleic acid that has an action of suppressing the expression of the target gene using RNA interference (RNAi).

The target gene in the present invention is not particularly limited as long as it is a gene that produces and expresses mRNA, but, for example, a gene related to tumor or inflammation is preferable, for example, vascular endothelial growth factor (hereinafter referred to as VEGF). Vascular endothelial growth factor receptor (vascular endothelial growth factor thereceptor, hereinafter abbreviated as VEGFR), fibroblast growth factor, fibroblast growth factor receptor, platelet-derived growth factor, platelet-derived growth factor receptor, liver Cell growth factor, hepatocyte growth factor receptor, Kruppel-like factor (KLF), express sequence tag (Ets) transcription factor, nuclear factor, hypoxia-inducible factor, cell cycle-related factor, chromosome Replication-related factor, chromosome repair-related factor, microtubule-related factor, growth signal pathway-related factor, growth-related transcription factor or apoptosis-related factor Examples include genes encoding proteins, specifically VEGF gene, VEGFR gene, fibroblast growth factor gene, fibroblast growth factor receptor gene, platelet-derived growth factor gene, platelet-derived growth factor receptor gene, Hepatocyte growth factor gene, hepatocyte growth factor receptor gene, KLF gene, Ets transcription factor gene, nuclear factor gene, hypoxia inducible factor gene, cell cycle related factor gene, chromosome replication related factor gene, chromosome repair related factor gene, Examples include a microtubule-related factor gene (for example, CKAP5 gene), a growth signal pathway-related factor gene (for example, KRAS gene), a growth-related transcription factor gene, or an apoptosis-related factor (for example, BCL-2 gene).

As the target gene in the present invention, for example, a gene expressed in the liver, lung, kidney or spleen is preferable, and a gene expressed in the liver is more preferable. For example, the above-mentioned gene related to tumor or inflammation, hepatitis B virus genome , Hepatitis C virus genome, apolipoprotein (APO), hydroxymethylglutaryl (HMG) CoA reductase, kexin type 9 serine protease (PCSK9), factor 12, glucagon receptor, glucocorticoid receptor, leukotriene receptor, Thromboxane A2 receptor, histamine H1 receptor, carbonic anhydrase, angiotensin converting enzyme, renin, p53, tyrosine phosphatase (PTP), sodium-dependent glucose transporter, tumor necrosis factor, interleukin, hepcidin, transsilence, antithrombin , Protein C or Matriptase enzymes (e.g., TMPRSS6 gene, etc.) and the gene and the like which encodes a protein, such as.

Examples of the nucleic acid that suppresses the expression of the target gene include, for example, a nucleotide sequence that is complementary to the partial nucleotide sequence of the mRNA of the gene encoding the protein (target gene) and suppresses the expression of the target gene. If it is a nucleic acid, for example, any nucleic acid such as a double-stranded nucleic acid such as siRNA (short (interference RNA) or miRNA (micro RNA), a single-stranded nucleic acid such as shRNA (short hairpin RNA), antisense nucleic acid or ribozyme, etc. Although it may be used, a double-stranded nucleic acid is preferred.

A nucleic acid containing a base sequence complementary to a part of the base sequence of the target gene mRNA is called an antisense strand nucleic acid, and a nucleic acid containing a base sequence complementary to the base sequence of the antisense strand nucleic acid is a sense strand. Also called nucleic acid. A sense strand nucleic acid refers to a nucleic acid capable of forming a double strand forming part by pairing with an antisense strand nucleic acid, such as a nucleic acid itself consisting of a partial base sequence of a target gene.

A double-stranded nucleic acid refers to a nucleic acid in which two strands are paired and have a duplex forming part. The double-stranded forming part refers to a part where nucleotides constituting the double-stranded nucleic acid or a derivative thereof constitute a base pair to form a double strand. The base pair constituting the duplex forming part is usually 15 to 27 base pairs, preferably 15 to 25 base pairs, more preferably 15 to 23 base pairs, further preferably 15 to 21 base pairs, and 15 to 19 base pairs. Base pairs are particularly preferred.

As the antisense strand nucleic acid of the duplex forming part, for example, a nucleic acid comprising a partial sequence of the mRNA of the target gene, or 1 to 3 bases, preferably 1 to 2 bases, more preferably 1 base in the nucleic acid is substituted. A nucleic acid that is deleted or added and has the activity of suppressing the expression of the target protein is preferably used. The single-stranded nucleic acid constituting the double-stranded nucleic acid usually consists of a series of 15 to 30 bases (nucleosides), preferably 15 to 29 bases, more preferably 15 to 27 bases, and further preferably 15 to 25 bases. 17 to 23 bases are particularly preferred, and 19 to 21 bases are most preferred.

The antisense strand, the sense strand, or both of the nucleic acids constituting the double-stranded nucleic acid have an additional nucleic acid that does not form a duplex on the 3 ′ side or 5 ′ side following the duplex forming part. May be. The part that does not form a double chain is also referred to as a protrusion (overhang).

As the double-stranded nucleic acid having an overhang, for example, one having an overhang of 1 to 4 bases, usually 1 to 3 bases at the 3 ′ end or 5 ′ end of at least one strand is used. Those having a protruding portion made of a base are preferably used, and those having a protruding portion made of dTdT or UU are more preferably used. The overhang can have only the antisense strand, only the sense strand, and both the antisense strand and the sense strand, but a double-stranded nucleic acid having an overhang on both the antisense strand and the sense strand is preferably used. .

Sequence that matches part or all of the base sequence of the target gene mRNA following the duplex formation part, or part or all of the base sequence of the complementary strand of the target gene mRNA following the duplex formation part Sequences that match may be used. Furthermore, as a nucleic acid that suppresses the expression of a target gene, for example, a nucleic acid molecule that generates the above double-stranded nucleic acid by the action of a ribonuclease such as Dicer (International Publication No. 2005/089287), a 3 ′ end or a 5 ′ A double-stranded nucleic acid or the like that does not have a terminal protruding portion can also be used.

When the double-stranded nucleic acid is siRNA, preferably the antisense strand has a sequence of at least the 1st to 17th bases (nucleosides) from the 5 ′ end to the 3 ′ end, and the mRNA of the target gene. It is a sequence of bases complementary to a continuous 17-base sequence, and more preferably, the antisense strand has a sequence of the 1st to 19th bases from the 5 ′ end to the 3 ′ end, and the target gene The base sequence is complementary to the 19-base sequence of mRNA, or the base sequence 1 to 21 is the base sequence complementary to the 21-base sequence of the target gene mRNA. Alternatively, the sequence of the 1st to 25th bases is a sequence of bases complementary to the sequence of 25 consecutive bases of the mRNA of the target gene.

Furthermore, when the nucleic acid used in the present invention is siRNA, preferably 10 to 70%, more preferably 15 to 60%, and still more preferably 20 to 50% of the sugar in the nucleic acid is modified at the 2 ′ position. Ribose substituted with In the present invention, ribose substituted with a modifying group at the 2′-position means that the hydroxyl at the 2′-position of ribose is substituted with the modifying group, and the configuration is the same as the hydroxy at the 2′-position of ribose. Although it may be present or different, the configuration is preferably the same as that of the 2′-hydroxy of ribose. Examples of the modifying group in ribose substituted with a modifying group at the 2′-position include those exemplified in the definition of the modifying group in the 2′-modified nucleotide in the sugar moiety-modified nucleotide and the hydrogen atom, such as 2′-cyano, 2 ′ -Halogen, 2'-O-cyano, 2'-alkyl, 2'-substituted alkyl, 2'-O-alkyl, 2'-O-substituted alkyl, 2'-O-alkenyl, 2'-O-substituted alkenyl , 2'-Se-alkyl, 2'-Se-substituted alkyl, etc. are preferred, 2'-cyano, 2'-fluoro, 2'-chloro, 2'-bromo, 2'-trifluoromethyl, 2'-O -Methyl, 2'-O-ethyl, 2'-O-isopropyl, 2'-O-trifluoromethyl, 2'-O- [2- (methoxy) ethyl], 2'-O- (3-aminopropyl ), 2'-O- [2- (N, N-dimethyl) aminooxy] ethyl, 2'-O- [3- (N, N-dimethylamino) propyl], 2'-O- {2- [ 2- (N, N-dimethylamino) ethoxy] ethyl}, 2'-O- [2- (methyl Amino) -2-oxoethyl], 2'-Se-methyl, hydrogen atom and the like are more preferable, 2'-O-methyl, 2'-O-ethyl, 2'-fluoro, hydrogen atom and the like are more preferable, and 2'- -O-methyl, 2'-O-fluoro is most preferred.

The nucleic acid used in the present invention includes a derivative in which an oxygen atom or the like contained in a phosphoric acid part, an ester part or the like in the structure of the nucleic acid is substituted with another atom such as a sulfur atom.

The sugar that binds to the 5 'terminal base of the antisense strand and the sense strand has a 5'-positioned hydroxy group, either a phosphate group or the above-mentioned modifying group, or an in vivo nucleolytic enzyme, etc. It may be modified by a group that is converted to a modifying group.

The sugar that binds to the base at the 3 ′ end of the antisense strand and the sense strand is such that the 3′-position hydroxy is a phosphate group or the above-mentioned modifying group, or an in vivo nucleolytic enzyme, etc. It may be modified by a group that is converted to a modifying group.

As the single-stranded nucleic acid, for example, 15 to 27 bases (nucleoside) of the target gene, preferably 15 to 25 bases, more preferably 15 to 23 bases, still more preferably 15 to 21 bases, particularly preferably 15 A nucleic acid comprising a sequence complementary to a sequence consisting of ˜19 bases, or 1 to 3 bases, preferably 1 to 2 bases, more preferably 1 base is substituted, deleted or added in the nucleic acid, and the target protein expression inhibitory activity Any nucleic acid may be used as long as it has a nucleic acid. The single-stranded nucleic acid is preferably composed of a series of 10 to 30 bases (nucleosides), more preferably 10 to 27 bases, further preferably 10 to 25 bases, particularly preferably 10 to 23 bases. Nucleic acids are preferably used.

As the single-stranded nucleic acid, one obtained by linking the antisense strand and the sense strand constituting the double-stranded nucleic acid via a spacer sequence (spacer oligonucleotide) may be used. The spacer oligonucleotide is preferably a 6- to 12-base single-stranded nucleic acid molecule, and the sequence at the 5 'end is preferably 2 Us. An example of a spacer oligonucleotide is a nucleic acid having the sequence UUCAAGAGA. Either the antisense strand or the sense strand connected by the spacer oligonucleotide may be on the 5 'side. The single-stranded nucleic acid is preferably, for example, a single-stranded nucleic acid such as shRNA having a duplex forming part by a stem-loop structure. Single-stranded nucleic acids such as shRNA are usually 50 to 70 bases in length.

A nucleic acid having a length of 70 bases or less, preferably 50 bases or less, more preferably 30 bases or less, designed to produce the above single-stranded nucleic acid or double-stranded nucleic acid by the action of ribonuclease or the like may be used. Good.

The nucleic acid used in the present invention can be obtained using a known RNA or DNA synthesis method and RNA or DNA modification method.

The nucleic acid-containing lipid nanoparticles of the present invention are a complex of lipid A and nucleic acid, but lipid A may contain one kind or two or more kinds.
The nucleic acid-containing lipid nanoparticles of the present invention contain, in addition to lipid A and nucleic acid, one or more lipids B other than lipid A, neutral lipids and / or lipid derivatives or fatty acid derivatives of water-soluble polymers. You may do it.

The nucleic acid-containing lipid nanoparticles of the present invention may contain one or more lipid B together with lipid A.

The nucleic acid-containing lipid nanoparticles of the present invention can contain not only nucleic acids but also compounds that are chemically similar to nucleic acids (anionic polymers such as anionic peptides).

In the present invention, the nucleic acid comprises lipid A and, if necessary, other lipids (lipid derivatives or fatty acid derivatives of water-soluble polymers, neutral lipids, and / or one amino group which may be substituted or one It is dissolved in a water-miscible organic solvent together with a hydrophilic part having a quaternary ammonium group and a lipid (lipid B) containing a hydrophobic part having two independent hydrocarbon groups which may be substituted in the same molecule. (First lipid solution). In the preparation of the first lipid solution, the nucleic acid may be dissolved in water or an aqueous buffer solution and added to the lipid organic solvent solution, or the lipid organic solvent solution may be added to the nucleic acid water or buffer solution. Also good. Furthermore, an organic solvent solution of lipid may be added to the lyophilized nucleic acid.

Nucleic acid, lipid A, and other lipids as necessary (lipid derivatives or fatty acid derivatives of water-soluble polymers, neutral lipids, and / or one amino group or one quaternary ammonium group that may be substituted) An organic solvent solution (first lipid solution) was prepared once with a lipid (lipid B)) containing a hydrophilic part having a hydrophobic part and a hydrophobic part having two independent hydrocarbon groups that may be substituted in the same molecule. Later, a third lipid solution may be prepared by adding an organic solvent solution (second lipid solution) containing a lipid derivative or fatty acid derivative of a water-soluble polymer.

In the present invention, the first or third lipid solution is mixed with water or an aqueous buffer solution. At this time, lipid nanoparticles having a small size can be obtained without agglomeration by rapidly reducing the organic solvent concentration.
In mixing the first or third lipid solution with water or an aqueous buffer solution, the former may be added to the latter, or the latter may be added to the former. Moreover, you may add the former and the latter simultaneously to a container, stirring. Furthermore, the former and the latter can be mixed in-line. In this case, for example, a T-connector or the like can be used as the in-line mixing device.

The average particle size of the nucleic acid-containing lipid nanoparticles of the present invention is the nucleic acid to be used, a lipid having one quaternary ammonium group and a lipid (lipid A) having three independent hydrocarbon groups that may be substituted, and Although it is influenced by other lipids, it can be freely controlled by various parameters in the production process. A person skilled in the art prepares a particle sample by appropriately changing various parameters in the production process necessary to control the average particle size of the nucleic acid-containing lipid nanoparticles of the present invention, and the average particle size of the obtained sample Can be determined by measuring. Parameters necessary for controlling the average particle diameter include the nucleic acid concentration in the organic solvent solution, the concentration of each lipid, the temperature, the composition of the organic solvent, and the like. Parameters necessary for controlling the average particle size include temperature, the amount of water or aqueous buffer solution, the rate of addition of each solution during the dilution operation of the nucleic acid and lipid organic solvent solution with water or aqueous buffer solution, and the like. .

When not containing phosphatidylcholine (PC) and cholesterol (Chol), it has one quaternary ammonium group as a hydrophilic part in the organic solvent solution before being mixed with water or an aqueous buffer solution, and may be substituted The concentration of the lipid having three independent hydrocarbon groups (lipid A) is not particularly limited, but is preferably 1 to 2000 μM, more preferably 5 to 400 μM, still more preferably 10 to 200 μM, and 20 to 100 μM. Most preferred.

The concentration of the nucleic acid in the organic solvent solution before mixing with water or an aqueous buffer solution in the case of not containing PC and Chol is not particularly limited, but is preferably 0.03 to 15 μM, more preferably 0.15 to 3.0 μM, and more preferably 0.3 to 1.5 More preferred is μM.

The concentration of the lipid or fatty acid derivative of the water-soluble polymer in the organic solvent solution before mixing with water or an aqueous buffer solution in the case of not containing PC and Chol is not particularly limited, but is preferably 0.5 to 200 μM, 2.5 ˜40 μM is more preferable, and 5 to 20 μM is even more preferable.

The concentration of the cationic lipid in the organic solvent solution before mixing with water or an aqueous buffer solution in the case of not containing PC and Chol is not particularly limited, but is preferably 1 to 2000 μM, more preferably 5 to 400 μM, 10 More preferably, ˜200 μM is most preferable, and 20 to 100 μM is most preferable.

The total concentration of all lipids in the organic solvent solution before mixing with water or an aqueous buffer solution when not containing PC and Chol is not particularly limited, but is preferably 5 to 2000 μM, more preferably 25 to 400 μM 50 to 200 μM is more preferable.

Independent three hydrocarbons that have one quaternary ammonium group as the hydrophilic part in the organic solvent solution before mixing with water or buffer solution in the case of containing PC and Chol, and may be substituted The concentration of the group-containing lipid (lipid A) is preferably 0.2 to 1800 μM, more preferably 1 to 360 μM, further preferably 2 to 180 μM, and most preferably 5 to 100 μM.

In the case of containing PC and Chol, the concentration of the nucleic acid in the organic solvent solution before being mixed with water or an aqueous buffer solution is preferably 0.02 to 45 μM, more preferably 0.1 to 10 μM, further preferably 0.2 to 5 μM, and 0.3 ˜3 μM is most preferred.

In the case of containing PC and Chol, the concentration of the water-soluble polymer lipid derivative or fatty acid derivative in the organic solvent solution before being mixed with water or an aqueous buffer solution is preferably 0.3 to 1000 μM, more preferably 1.5 to 200 μM. More preferably, 3 to 100 μM is more preferable, and 5 to 50 μM is most preferable.

In the case of containing PC and Chol, the concentration of the cationic lipid in the organic solvent solution before being mixed with water or an aqueous buffer solution is preferably 2.5 to 4200 μM, more preferably 12.5 to 840 μM, and 25 to 420 μM. More preferred is 50 to 210 μM.

In the case of containing PC and Chol, the concentration of neutral lipid in the organic solvent solution before mixing with water or an aqueous buffer solution is preferably 2.5 to 5000 μM, more preferably 12.5 to 1000 μM, and further preferably 25 to 500 μM. 50 to 250 μM is most preferable.

In the case of containing PC and Chol, the combined concentration of all lipids in the organic solvent solution before being mixed with water or an aqueous buffer solution is preferably 10 to 8000 μM, more preferably 50 to 1600 μM, and more preferably 100 to 800 μM is more preferable, and 150 to 400 μM is most preferable.

The temperature at which the organic solvent solution containing the nucleic acid and lipid is prepared is not particularly limited as long as the nucleic acid and lipid are dissolved, but is preferably 10 to 60 ° C, more preferably 20 to 50 ° C, and further preferably 20 to 30 ° C. preferable. In addition, when heating at 30 degreeC or more, the solubility of a nucleic acid and a lipid increases and a lipid nanoparticle can be manufactured with a smaller solvent amount.

Although it does not specifically limit as an organic solvent in the organic solvent solution containing a nucleic acid and a lipid, C1-C6 alcohols, such as methanol, ethanol, a propanol, a butanol, etc. which contain 0-50% (v / v) water, or these mixtures More preferred is ethanol or propanol containing 0 to 50% (v / v) water, and even more preferred is ethanol containing 0 to 50% (v / v) water. Here, “% (v / v)” indicates the volume percentage of the solute in the total volume of the solution, and so on.

An inorganic acid such as hydrochloric acid, acetic acid, phosphoric acid, or a salt of these acids can be added to the solvent in the organic solvent solution containing nucleic acid and lipid. In this case, the pH of the solvent is preferably 1 to 7, more preferably 1 to 5, and further preferably 2 to 4.

In the operation of adding water or an aqueous buffer solution to an organic solvent solution containing nucleic acid and lipid, the volume of water or aqueous buffer solution used is not particularly limited, but is 0.5% relative to the volume of the organic solvent solution of nucleic acid and lipid. ~ 100 times is preferable, 1.5 to 20 times is more preferable, and 2.0 to 10 times is more preferable.

In this case, the concentration of the organic solvent after adding water or an aqueous buffer solution is not particularly limited, but is preferably 50% (v / v) or less, more preferably 40% (v / v) or less with respect to the obtained solution. Preferably, it is more preferably 30% (v / v) or less, and most preferably 20% / (v / v) or less. The aqueous buffer solution is not particularly limited as long as it has a buffering action, and examples thereof include a phosphate buffer aqueous solution, a citrate buffer aqueous solution, and an acetate buffer aqueous solution.

The temperature at the time of the above addition operation is not particularly limited, but is preferably 10 to 60 ° C, more preferably 20 to 50 ° C, and further preferably 20 to 30 ° C.

In the above addition operation, it is important to quickly lower the organic solvent solution. Specifically, it is preferable to change the organic solvent concentration from 70% 70 (v / v) to 50% (v / v) within 1 minute, more preferably within 0.5 minutes, 0.1% More preferably, it is changed within minutes, and most preferably within 0.05 minutes.

In the nucleic acid-containing lipid nanoparticle of the present invention, the total number of lipid A molecules is not particularly limited, but the number of moles of the quaternary ammonium group in the lipid A is the phosphorus atom of the nucleic acid constituting the nucleic acid-containing lipid nanoparticle of the present invention. The molar amount is preferably 0.01 times the molar amount or more, more preferably 0.1 to 10 times the molar amount, even more preferably 0.1 to 4 times the molar amount, and more preferably 0.1 to 2 times the molar amount. Even more preferably, it is most preferably 0.1 to 1 times the molar amount. When the nucleic acid-containing lipid nanoparticle contains lipid B, the total number of lipid B molecules is not particularly limited, but the number of moles of lipid B is equal to the number of moles of phosphorus atoms of the nucleic acid constituting the nucleic acid-containing lipid nanoparticle of the present invention. It is preferably 0.1 to 10 times the molar amount, more preferably 0.5 to 9 times the molar amount, further preferably 1 to 8 times the molar amount, and 1.5 to 6 times the molar amount. Most preferred.

When lipid B is contained in the nucleic acid-containing lipid nanoparticles of the present invention, the ratio of the number of moles of lipid A to the number of moles of lipid B (number of moles of lipid A / number of moles of lipid B) is 0.001 or more. Preferably, it is 0.003 to 10, more preferably 0.005 to 5, still more preferably 0.01 to 3, and most preferably 0.01 to 2.

In the nucleic acid-containing lipid nanoparticles of the present invention, the ratio of the total number of moles of lipids to the number of moles of nucleic acids (the total number of moles of lipids / number of moles of nucleic acids) is preferably 50 or more, and preferably 100 to 1000. More preferably, it is more preferably from 120 to 800, even more preferably from 140 to 600, and most preferably from 200 to 500.

When the nucleic acid-containing lipid nanoparticle of the present invention contains lipid B, the total number of lipid B molecules in the nucleic acid-containing lipid nanoparticle is not particularly limited, but is 0.1 times the molar amount or more relative to the total number of moles of lipid. Preferably, it is 0.15 times the molar amount or more, more preferably 0.2 times the molar amount or more, and even more preferably 0.25 times the molar amount or more. Further, the total number of lipid B molecules in the nucleic acid-containing lipid nanoparticles is not particularly limited, but is preferably 0.7 times the molar amount or less, more preferably 0.65 times the molar amount or less with respect to the total number of moles of lipid. More preferably, it is 0.6 times the molar amount or less.
The total number of lipid B molecules in the nucleic acid-containing lipid nanoparticles is more preferably 0.10 to 0.70 times the molar amount of the total lipid in the combination of the above upper and lower limit preferable ranges, 0.15 to The molar amount is more preferably 0.65 times, even more preferably 0.20 to 0.65 times the molar amount, and most preferably 0.25 to 0.60 times the molar amount.

The neutral lipid may be any of simple lipids, complex lipids or derived lipids, and examples thereof include, but are not limited to, phospholipids, glyceroglycolipids, sphingoglycolipids, sphingoids, and sterols. Further, neutral lipids may be used alone or in combination of two or more.

When the nucleic acid-containing lipid nanoparticles of the present invention contain neutral lipids, the total number of neutral lipid molecules is not particularly limited, but is 0.10 to 0.75 times the molar amount of total lipids. The molar amount is preferably 0.20 to 0.70, more preferably 0.20 to 0.65, and most preferably 0.30 to 0.60.

Examples of the phospholipid in the neutral lipid include phosphatidylcholine (PC) (specifically soybean phosphatidylcholine, egg yolk phosphatidylcholine (EPC), distearoylphosphatidylcholine, 1,2-distearoyl-sn-glycero-3-phosphocholine (DSPC)). , Dipalmitoyl phosphatidylcholine, 1,2-dipalmitoyl-sn-glycero-3-phosphocholine (DPPC), palmitoyl oleoyl phosphatidylcholine (POPC), dimyristoyl phosphatidylcholine (DMPC), dioleoylphosphatidylcholine (DOPC), etc.), phosphatidylethanol Amines (specifically distearoylphosphatidylethanolamine (DSPE), dipalmitoylphosphatidylethanolamine (DPPE), dioleoylphosphatidylethanolamine (DOPE), dimyristoyl phosphoethanolamine (DMPE), 16-0-mono Chill PE, 16-0-dimethyl PE, 18-1-trans PE, palmitoyl oleoyl-phosphatidylethanolamine (POPE), 1 -stearoyl-2-oleoyl-phosphatidylethanolamine (SOPE), etc.), glycerophospholipid (specific) Phosphatidylserine, phosphatidic acid, phosphatidylglycerol, phosphatidylinositol, palmitoyloleoylphosphatidylglycerol (POPG), lysophosphatidylcholine, etc.), sphingophospholipids (specifically sphingomyelin, ceramide phosphoethanolamine, ceramide phosphoglycerol, ceramide) Phosphoglycerophosphate, etc.), glycerophosphonolipid, sphingophosphonolipid, natural lecithin (specifically egg yolk lecithin, soybean lecithin, etc.) or hydrogenated phospholipid (specifically hydrogenated soybean phosphatidyl). While natural phospholipids or synthetic phosphorus and the like), and the like without limitation.

Examples of the glyceroglycolipid in the neutral lipid include, but are not limited to, sulfoxyribosyl glyceride, diglycosyl diglyceride, digalactosyl diglyceride, galactosyl diglyceride, glycosyl diglyceride and the like.

Examples of the glycosphingolipid in the neutral lipid include, but are not limited to, galactosyl cerebroside, lactosyl cerebroside, ganglioside, and the like.

Examples of the sphingoid in the neutral lipid include, but are not limited to, sphingan, icosasphingan, sphingosine, and derivatives thereof. . As the derivative, for example, —NH ₂ such as sphingan, icosasphingan or sphingosine —NHCO (CH ₂ ) xCH ₃ (wherein x is an integer of 0 to 18, among which 6, 12 or 18 is preferable. However, it is not limited to these.

Examples of sterols in neutral lipids include cholesterol (Chol), dihydrocholesterol, lanosterol, β-sitosterol, campesterol, stigmasterol, brassicasterol, ergocasterol, fucostosterol, or 3β- [N- (N ′, N'-dimethylaminoethyl) carbamoyl] cholesterol (DC-Chol) and the like, but are not limited thereto.

Examples of the macromolecule include protein, albumin, dextran, polyfect, chitosan, dextran sulfate, such as poly-L-lysine, polyethyleneimine, polyaspartic acid, styrene maleic acid copolymer, isopropylacrylamide-acrylpyrrolidone. Examples include, but are not limited to, a copolymer, a polyethylene glycol-modified dendrimer, polylactic acid, a polymer such as polylactic acid polyglycolic acid or polyethylene glycolated polylactic acid, or micelles composed of one or more salts thereof.

Here, the polymer salt includes, for example, metal salts, ammonium group salts, acid addition salts, organic amine addition salts, amino acid addition salts, and the like. Examples of the metal salt include, but are not limited to, alkali metal salts such as lithium salt, sodium salt and potassium salt, alkaline earth metal salts such as magnesium salt and calcium salt, aluminum salt and zinc salt. . Examples of ammonium group salts include, but are not limited to, salts such as ammonium groups or tetramethylammonium groups. . Examples of the acid addition salt include inorganic acid salts such as hydrochloride, sulfate, nitrate or phosphate, and organic acid salts such as acetate, maleate, fumarate or citrate. It is not limited to. . Examples of organic amine addition salts include, but are not limited to, addition salts such as morpholine or piperidine. . Examples of amino acid addition salts include, but are not limited to, addition salts such as glycine, phenylalanine, aspartic acid, glutamic acid, or lysine.

Any of the nucleic acid-containing lipid nanoparticles of the present invention may contain, for example, a lipid derivative or fatty acid derivative of one or more substances selected from sugars, peptides, nucleic acids, and water-soluble polymers, or a surfactant.

The lipid derivative or fatty acid derivative of one or more substances selected from sugars, peptides, nucleic acids, and water-soluble polymers, or the surfactant, preferably a lipid or fatty acid derivative of a glycolipid or a water-soluble polymer. More preferred are lipid derivatives or fatty acid derivatives of water-soluble polymers. A lipid derivative or fatty acid derivative of one or more substances selected from sugars, peptides, nucleic acids, and water-soluble polymers, or a surfactant is a part of the molecule and other components of the composition, such as hydrophobic affinity, static It is a substance with a two-sided property that has the property of binding by electrical interaction, etc., and the other part has the property of binding to the solvent at the time of production of the composition, for example, hydrophilic affinity, electrostatic interaction, etc. Preferably there is.

Examples of lipid derivatives or fatty acid derivatives of sugars, peptides or nucleic acids include sugars such as sucrose, sorbitol, and lactose, such as casein-derived peptides, egg white-derived peptides, soybean-derived peptides, peptides such as glutathione, or, for example, DNA, Nucleic acids such as RNA, plasmid, siRNA, ODN and the like and neutral lipids listed in the definition of the composition or fatty acids such as stearic acid, palmitic acid, myristic acid, lauric acid, etc. Is mentioned.

Examples of the sugar lipid derivative or fatty acid derivative include glyceroglycolipid and glycosphingolipid mentioned in the definition of the composition.

Examples of the water-soluble polymer lipid derivative or fatty acid derivative include polyethylene glycol, polyglycerin, polyethyleneimine, polyvinyl alcohol, polyacrylic acid, polyacrylamide, oligosaccharide, dextrin, water-soluble cellulose, dextran, chondroitin sulfate, and polyglycerin. , Chitosan, polyvinylpyrrolidone, polyaspartic acid amide, poly-L-lysine, mannan, pullulan, oligoglycerol etc. or their derivatives and neutral lipids mentioned in the above definition of the composition or eg stearic acid, palmitic Examples include those formed by bonding with fatty acids such as acid, myristic acid or lauric acid, and salts thereof, more preferably lipid derivatives such as polyethylene glycol or polyglycerin or fatty acid derivatives and Include those salts, more preferable example is a lipid derivative or fatty acid derivatives and salts thereof polyethylene glycol.

Examples of polyethylene glycol lipid derivatives or fatty acid derivatives include polyethylene glycolated lipids [specifically, polyethylene glycol-phosphatidylethanolamine (more specifically 1,2-distearoyl-sn-glycero-3-phosphoethanolamine). -N- [methoxy (polyethylene glycol) -2000] (PEG-DSPE), 1,2-dipalmitoyl-sn-glycero-3-phosphoethanolamine-N- [methoxy (polyethylene glycol) -2000] (PEG-DPPE ), 1,2-dimyristoyl-sn-glycero-3-phosphoethanolamine-N- [methoxy (polyethylene glycol) -2000] (PEG-DMPE), etc.), polyoxyethylene hydrogenated castor oil 60, Cremofoyer ( CREMOPHOR EL), polyethylene glycol sorbitan fatty acid esters (specifically polyoxyethylene sorbitan monooleate, etc.) or polyethylene glycol Call fatty acid esters and the like, and more preferably, polyethylene glycolated lipid.

Examples of the lipid derivative or fatty acid derivative of polyglycerin include polyglycerinized lipids (specifically polyglycerin-phosphatidylethanolamine) or polyglycerin fatty acid esters, and more preferably, polyglycerinized lipids. Can be mentioned.

Examples of the surfactant include polyoxyethylene sorbitan monooleate (specifically polysorbate 80 etc.), polyoxyethylene polyoxypropylene glycol (specifically Pluronic F68 etc.), sorbitan fatty acid ester (specifically Sorbitan monolaurate, sorbitan monooleate, etc.), polyoxyethylene derivatives (specifically polyoxyethylene hydrogenated castor oil 60, polyoxyethylene lauryl alcohol, etc.), glycerin fatty acid ester or polyethylene glycol alkyl ether, etc. Preferably, polyoxyethylene polyoxypropylene glycol, glycerin fatty acid ester, polyethylene glycol alkyl ether or the like is used.

In the nucleic acid-containing lipid nanoparticles of the present invention, the total number of water-soluble polymer lipid derivatives and fatty acid derivative molecules in the nucleic acid-containing lipid nanoparticles is not particularly limited, but is 0.005 times the molar amount or more of the total lipid moles. Preferably, it is 0.01 to 0.30 times molar amount, more preferably 0.02 to 0.25 times molar amount, still more preferably 0.03 to 0.20 times molar amount, and even more preferably 0.04 to 0.15 times molar amount. Even more preferred is an amount, and most preferred is a 0.04-0.12 molar amount.
In the present invention, the total lipid includes lipid A and lipid derivatives and fatty acid derivatives of water-soluble polymers, and optionally includes lipid B and neutral lipid. That is, the number of moles of lipid A is a double mole quantity of the lipid derivative and fatty acid derivative of the water-soluble polymer, and sometimes a double mole quantity of lipid B and The total of the double molar amount of neutral lipid is reduced by 1 to the double molar amount.

In addition, the nucleic acid-containing lipid nanoparticles of the present invention can be optionally subjected to surface modification with, for example, a water-soluble polymer [Radasic, edited by F. Martin, “Stealth liposomes”. [See Stealth Liposomes ”(USA), CRC Inc, 1995, p.93-102]. Examples of water-soluble polymers that can be used for surface modification include polyethylene glycol, polyglycerin, polyethyleneimine, polyvinyl alcohol, polyacrylic acid, polyacrylamide, oligosaccharide, dextrin, water-soluble cellulose, dextran, chondroitin sulfate, poly Examples include glycerin, chitosan, polyvinylpyrrolidone, polyaspartic acid amide, poly-L-lysine, mannan, pullulan, oligoglycerol, etc., preferably polyethylene glycol, polyglycerin, polyethyleneimine, polyvinyl alcohol, polyacrylic acid, polyacrylamide, etc. More preferably, polyethylene glycol, polyglycerin and the like can be mentioned, but not limited thereto. For the surface modification, a lipid derivative or fatty acid derivative (as defined above) of one or more substances selected from sugars, peptides, nucleic acids, and water-soluble polymers, or a surfactant or the like can be used. The surface modification is one of methods in which the lipid derivative or fatty acid derivative of one or more substances selected from sugars, peptides, nucleic acids, and water-soluble polymers, or a surfactant is contained in the nucleic acid-containing lipid nanoparticles of the present invention. One.

Optionally, the targeting ligand can be directly bound to the surface of the nucleic acid-containing lipid nanoparticle of the present invention by covalently binding to the polar head residue of the lipid component of the nucleic acid-containing lipid nanoparticle of the present invention ( (See International Publication No. 2006/116107).

The average particle size of the nucleic acid-containing lipid nanoparticles of the present invention can be further adjusted after preparation of the lipid nanoparticles. As a method for adjusting the average particle size, for example, an extrusion method, a method of mechanically pulverizing large multilamellar liposomes (MLV) or the like (specifically using a manton gourin, a microfluidizer, etc.) RHMuller, S. Benita, B. Bohm, “Emulsion and サ Nanosuspensions for the Emulsion and Nanosuspensions for the Formulation of Poorly Soluble Drugs) ”, Scientific Publishers Stuttgart, 1998, p.267-294].

The average size of the nucleic acid-containing lipid nanoparticles of the present invention is preferably 20 to 65 nm, and more preferably 30 to 60 nm.

The size of the nucleic acid-containing lipid nanoparticles of the present invention can be measured, for example, by a dynamic light scattering method.

The nucleic acid-containing lipid nanoparticles of the present invention can be introduced into cells by introducing the nucleic acid-containing lipid nanoparticles of the present invention into mammalian cells.

In vivo introduction of the nucleic acid-containing lipid nanoparticles of the present invention into mammalian cells may be carried out according to known transfection procedures that can be performed in vivo. For example, the nucleic acid-containing lipid nanoparticles of the present invention are intravenously administered to mammals including humans, so that the nucleic acid-containing lipid nanoparticles are delivered to, for example, tumors or inflamed organs or sites. Nucleic acids in the nucleic acid-containing lipid nanoparticles can be introduced. The organ or site where the tumor or inflammation has occurred is not particularly limited, and examples thereof include stomach, large intestine, liver, lung, spleen, pancreas, kidney, bladder, skin, blood vessel, and eyeball. In addition, by administering the nucleic acid-containing lipid nanoparticles of the present invention intravenously to mammals including human beings, for example, delivered to the liver, stomach, lung, kidney, pancreas and / or spleen, cells of the delivery organ or site The nucleic acid in the nucleic acid-containing lipid nanoparticles of the present invention can be introduced into the inside. Liver, lung, spleen and / or kidney cells can be normal cells, cells associated with tumors or inflammation or cells associated with other diseases.

If the nucleic acid in the nucleic acid-containing lipid nanoparticles of the present invention is a nucleic acid having a target gene expression inhibitory action utilizing RNA interference (RNAi), the target gene expression is suppressed in vivo in mammalian cells in vivo. Nucleic acid or the like can be introduced, and the expression of the target gene can be suppressed. The administration subject is preferably a human.

If the target gene in the nucleic acid-containing lipid nanoparticles of the present invention is, for example, a gene expressed in the liver, stomach, lung, kidney, pancreas and / or spleen, preferably a gene expressed in the liver, the nucleic acid-containing lipid of the present invention Lipid nanoparticles can be used as a therapeutic or prophylactic agent for diseases associated with the liver, stomach, lung, kidney, pancreas or spleen, preferably as a therapeutic or prophylactic agent for diseases associated with the liver. That is, the present invention also provides a method for treating diseases related to liver, stomach, lung, kidney, pancreas or spleen, wherein the nucleic acid-containing lipid nanoparticles of the present invention described above are administered to mammals. The administration subject is preferably a person, and more preferably a person suffering from a disease related to the liver, stomach, lung, kidney, pancreas or spleen.

The nucleic acid-containing lipid nanoparticles of the present invention are effective in suppressing target genes in an in vivo drug efficacy evaluation model for therapeutic or prophylactic agents for diseases such as diseases related to liver, stomach, lung, kidney, pancreas or spleen. It can also be used as a tool for verification.

The nucleic acid-containing lipid nanoparticles of the present invention are, for example, tissues or organs containing a nucleic acid in a biological component such as a blood component (for example, blood, gastrointestinal tract), stabilizing the nucleic acid, reducing side effects, or expressing a target gene. It can also be used as a preparation for the purpose of increasing drug accumulation.

When the nucleic acid-containing lipid nanoparticles of the present invention are used as a therapeutic or prophylactic agent for diseases related to the liver, lung, kidney or spleen of pharmaceuticals, the administration route is the most effective route for treatment. For example, parenteral administration or oral administration such as buccal, intratracheal, rectal, subcutaneous, intramuscular or intravenous administration can be mentioned, preferably intravenous administration, subcutaneous administration or intramuscular administration. More preferably, intravenous administration is mentioned.

The dose varies depending on the disease state, age, administration route, etc. of the administration subject, but for example, it may be administered so that the daily dose converted to nucleic acid is about 0.1 μg to 1000 mg.

As a preparation suitable for intravenous administration or intramuscular administration, for example, an injection can be mentioned, and a dispersion of the composition prepared by the above-described method can be used as it is, for example, in the form of an injection or the like. The dispersion can be used after removing the solvent, for example, by filtration, centrifugation, etc., or the dispersion can be used after lyophilization, and / or, for example, mannitol, lactose, trehalose, maltose, glycine, etc. The dispersion added with the excipient can also be lyophilized for use.

In the case of injections, for example, water, acid, alkali, various buffers, physiological saline or amino acid infusion, etc. are mixed with the dispersion of the composition or the composition after removing or lyophilizing the solvent. It is preferable to prepare an injection. Further, for example, an injection can be prepared by adding an antioxidant such as citric acid, ascorbic acid, cysteine or EDTA or an isotonic agent such as glycerin, glucose or sodium chloride. Further, for example, it can be cryopreserved by adding a cryopreservative such as glycerin.

[Example]
Next, the present invention will be specifically described with reference to Examples, Reference Examples, Comparative Examples, and Test Examples. However, the present invention is not limited to these examples, reference examples, comparative examples, and test examples.
The proton nuclear magnetic resonance spectra ( ¹ H NMR) shown in the Examples and Reference Examples were measured at 270 MHz, 300 MHz, or 400 MHz, and exchangeable protons were clearly observed depending on the compound and measurement conditions. It may not be done. In addition, although what is used normally is used as the notation of the multiplicity of signals, br represents an apparently wide signal.

N-methyl-2- (oleoyloxy) -N, N-bis (2- (oleoyloxy) ethyl) ethanaminium chloride (Compound I-1)
Process 1
To a solution of triethanolamine (Sigma-Aldrich, 0.115 g, 0.771 mmol) in chloroform (5 mL), oleic acid (Tokyo Chemical Industry, 0.784 g, 2.78 mmol), 1-ethyl-3- (3-Dimethylaminopropyl) carbodiimide hydrochloride (Tokyo Chemical Industry, 0.591 g, 3.08 mmol), Triethylamine (0.430 mL, 3.08 mmol) and N, N-dimethylaminopyridine (Nacalai Tesque, 0.024 g, 0.19) mmol) was added and stirred at room temperature overnight. Water was added to the reaction solution and extracted with chloroform. The organic layer was washed with a saturated aqueous sodium hydrogen carbonate solution and then with saturated brine, dried over anhydrous magnesium sulfate, filtered, and concentrated under reduced pressure. The obtained residue was purified by silica gel column chromatography (chloroform / hexane = 50/50 to 100/0) to give 2,2 ′, 2 ″ -nitrilotris (ethane-2,1-diyl) trioleate (0.439 g, 0.466 mmol, yield 60%).
ESI-MS m / z: 943 (M + H) ⁺ ; ¹ H-NMR (CDCl ₃ ) δ: 0.88 (t, J = 6.9 Hz, 9H), 1.23-1.36 (m, 60H), 1.58-1.63 ( m, 6H), 1.98-2.04 (m, 12H), 2.29 (t, J = 7.6 Hz, 6H), 2.83 (t, J = 6.1 Hz, 6H), 4.11 (t, J = 6.1 Hz, 6H), 5.31-5.38 (m, 6H).
Process 2
2,2 ', 2''-Nitrilotris (ethane-2,1-diyl) trioleate (0.439 g, 0.466 mmol) obtained in Step 1 was mixed with methyl iodide (Tokyo Chemical Industry Co., Ltd., 3 mL). The mixture was further stirred at room temperature overnight. The reaction solution was concentrated under reduced pressure. Dissolve the residue in a small amount of methanol-chloroform (1: 1) ion exchange resin (Dow Chemical, Dowex (TM) 1x-2 100 mesh, Cl type, about 20 times the amount, water and methanol And pre-washed with methanol and eluted with methanol-chloroform (1: 1). The eluate was concentrated under reduced pressure, and the resulting residue was purified by silica gel column chromatography (chloroform / methanol = 100/0 to 88/12) to give the title compound (0.342 g, 0.344 mmol, 74% yield). Obtained.
ESI-MS m / z: 957 (M) ⁺ ; ¹ H-NMR (CDCl ₃ ) δ: 0.88 (t, J = 6.9 Hz, 9H), 1.25-1.35 (m, 60H), 1.59-1.63 (m, 6H), 1.99-2.03 (m, 12H), 2.35 (t, J = 7.6 Hz, 6H), 3.56 (s, 3H), 4.21 (t, J = 4.9 Hz, 6H), 4.61 (t, J = 4.9 Hz, 6H), 5.30-5.38 (m, 6H).

N-methyl-2-((9Z, 12Z) -octadeca-9,12-dienoyloxy) -N, N-bis (2-((9Z, 12Z) -octadeca-9,12-dienoyloxy) ethyl) ethaneaminium Chloride (Compound I-2)
In the same manner as in Example 1, using (9Z, 12Z) -octadeca-9,12-dienoic acid (Sigma-Aldrich, 0.704 g, 2.51 mmol) instead of oleic acid, the title compound (0.100 g Through yield 22%).
ESI-MS m / z: 950 (M) ⁺ ; ¹ H-NMR (CDCl ₃ ) 0.89 (t, J = 7.0 Hz, 9H), 1.25-1.40 (m, 42H), 1.55-1.66 (m, 6H) , 2.05 (q, J = 6.9 Hz, 12H), 2.35 (t, J = 7.6 Hz, 6H), 2.77 (t, J = 6.3 Hz, 6H), 3.54 (s, 3H), 4.21 (t, J = 5.1 Hz, 6H), 4.59 (br s, 6H), 5.28-5.43 (m, 12H).

(9Z, 12Z) -N-Methyl-N, N-di ((9Z, 12Z) -octadeca-9,12-dienyl) octadeca-9,12-dien-1-aminium chloride (Compound I-3)
Process 1
Ammonia (Tokyo Chemical Industry Co., Ltd., approx. 7 mol / L methanol solution, 8.00 mL, 56.0 mmol) to (9Z, 12Z) -octadeca-9,12-dienyl methanesulfonate (Nu-Chek Ink (Nu-Chek Prep, Inc), 3.55 g, 10.1 mmol) was added, and the mixture was stirred at 130 ° C. for 3 hours using a microwave reactor. A saturated aqueous sodium hydrogen carbonate solution was added to the reaction mixture, and the mixture was extracted 5 times with chloroform. The organic layers were combined, washed with saturated brine, dried over anhydrous magnesium sulfate, filtered, and concentrated under reduced pressure to obtain a crude product of (9Z, 12Z) -octadeca-9,12-dien-1-amine. .
(9Z, 12Z) -octadeca-9,12-dienyl methanesulfonate (2.78 g, 8.07 mmol) and 50% aqueous sodium hydroxide solution (2.00 mL, 50.0 mmol) were added to the resulting crude product on an oil bath. The mixture was stirred at 110 ° C for 60 minutes. After cooling to room temperature, the reaction mixture was diluted with ethyl acetate, washed with water and then with saturated brine, dried over anhydrous magnesium sulfate, filtered, and concentrated under reduced pressure. The obtained residue was purified by silica gel column chromatography (hexane / ethyl acetate = 100/0 to 90/10) to give (9Z, 12Z) -tri (9Z, 12Z) -octadeca-9,12-dienylamine ( 1.09 g, 1.43 mmol, 18% yield).
ESI-MS m / z: 763 (M + H) ⁺ .
Process 2
Example 1 In the same manner as in Step 2, (9Z, 12Z) -tri (2) obtained in Step 1 instead of 2,2 ', 2''-nitrilotris (ethane-2,1-diyl) trioleate The title compound (1.06 g, 1.30 mol, 94% yield) was obtained using 9Z, 12Z) -octadeca-9,12-dienylamine (1.05 g, 1.38 mol).
ESI-MS m / z: 777 (M) ⁺ ; ¹ H-NMR (CDCl ₃ ) δ: 0.89 (t, J = 7.1 Hz, 9H), 1.22-1.45 (m, 48H), 1.61-1.69 (m, 6H), 2.05 (q, J = 6.8 Hz, 12H), 2.77 (t, J = 6.5 Hz, 6H), 3.35 (s, 3H), 3.44-3.50 (m, 6H), 5.29-5.42 (m, 12H ).

(Z) -N-Methyl-N, N-di ((Z) -octadeca-9-enyl) octadeca-9-en-1-aminium chloride (Compound I-4)
In the same manner as in Example 3, instead of (9Z, 12Z) -octadeca-9,12-dienyl methanesulfonate, (Z) -octadeca-9-enyl methanesulfonate (manufactured by Nu-Chek Prep, Inc.) To give the title compound (0.410 g, 0.501 mmol, overall yield 24%).
ESI-MS m / z: 783 (M) ⁺ ; ¹ H-NMR (CDCl ₃ ) δ: 0.88 (t, J = 6.9 Hz, 9H), 1.22-1.44 (m, 66H), 1.62-1.69 (m, 6H), 1.98-2.04 (m, 12H), 3.35 (s, 3H), 3.45-3.51 (m, 6H), 5.30-5.39 (m, 6H).

(11Z, 14Z) -N, N-di ((11Z, 14Z) -icosa-11,14-dienyl) -N-methylicosa-11,14-dien-1-aminium chloride (Compound I-5)
In the same manner as in Example 3, instead of (9Z, 12Z) -octadeca-9,12-dienyl methanesulfonate, (11Z, 14Z) -icosa-11,14-dienyl methanesulfonate (Nu-Chek Prep, Inc.) to give the title compound (0.323 g, 0.360 mmol, overall yield 25%).
¹ H-NMR (CDCl ₃ ) δ: 0.89 (t, J = 6.9 Hz, 9H), 1.24-1.43 (m, 63H), 1.61-1.69 (m, 6H), 2.05 (q, J = 6.8 Hz, 12H ), 2.77 (t, J = 6.6 Hz, 6H), 3.35 (s, 3H), 3.45-3.50 (m, 6H), 5.30-5.42 (m, 12H).

(9Z, 12Z) -N- (3-Hydroxypropyl) -N, N-di ((9Z, 12Z) -octadeca-9,12-dienyl) octadeca-9,12-dien-1-aminium chloride (Compound I -6)
To a solution of tri ((9Z, 12Z) -octadeca-9,12-dienyl) amine (0.199 g, 0.261 mmol) obtained in Step 1 of Example 3 in chloroform (0.3 mL), 3-iodopropan-1-ol ( Wako Pure Chemical Industries, Ltd., 0.194 g, 1.04 mmol) was added, and the mixture was reacted at 130 ° C. for 40 minutes in a microwave reactor. Dissolve the reaction solution in a small amount of ethanol and load it onto an ion exchange resin (Sigma-Aldirch, Amberlite® IRA-400, Cl type, approximately 20 times volume, pre-washed with water and ethanol) and elute with ethanol. did. The eluate was concentrated under reduced pressure, and the resulting residue was purified by silica gel column chromatography (chloroform / methanol = 100/0 to 88/12) to give the title compound (0.146 g, 0.170 mmol, yield 65%). Obtained.
ESI-MS m / z: 821 (M) ⁺ ; ¹ H-NMR (CDCl ₃ ) δ: 0.89 (t, J = 7.0 Hz, 9H), 1.27-1.39 (m, 49H), 1.67-1.74 (m, 6H), 1.93-1.99 (m, 2H), 2.05 (q, J = 6.9 Hz, 12H), 2.77 (t, J = 6.2 Hz, 6H), 3.14-3.19 (m, 6H), 3.70-3.74 (m , 2H), 3.79 (t, J = 5.1 Hz, 2H), 5.29-5.42 (m, 12H).

(9Z, 12Z) -N- (2-hydroxyethyl) -N, N-di ((9Z, 12Z) -octadeca-9,12-dienyl) octadeca-9,12-dien-1-aminium chloride (Compound I -7)
In the same manner as in Example 6, using 2-iodoethane-1-ol (manufactured by Tokyo Chemical Industry Co., Ltd.) instead of 3-iodopropan-1-ol, the title compound (0.211 g, 0.250 mmol, yield 85) %).
ESI-MS m / z: 807 (M) ⁺ ; ¹ H-NMR (CDCl ₃ ) δ: 0.89 (t, J = 7.0 Hz, 9H), 1.27-1.40 (m, 49H), 1.64-1.71 (m, 6H), 2.05 (q, J = 6.8 Hz, 12H), 2.77 (t, J = 6.3 Hz, 6H), 3.36-3.41 (m, 6H), 3.53-3.56 (m, 2H), 4.08-4.12 (m , 2H), 5.29-5.42 (m, 12H).

N, N, N-trimethyl-1,3-bis ((9Z, 12Z) -octadeca-9,12-dienoyloxy) -2-(((9Z, 12Z) -octadeca-9,12-dienoyloxy) methyl) propane -2-Aminium chloride (Compound II-1)
Process 1
To a solution of 2- (dimethylamino) -2- (hydroxymethyl) propane-1,3-diol (Zylexa Pharma, 0.252 g, 1.69 mmol) in chloroform (10 mL) (9Z, 12Z) -Octadeca-9,12-dienoic acid (2.37 g, 8.45 mmol), 1-ethyl-3- (3-dimethylaminopropyl) carbodiimide hydrochloride (1.62 g, 8.45 mmol) and N, N-dimethylaminopyridine (0.206 g, 1.69 mmol) was added and stirred at 60 ° C. overnight. After cooling to room temperature, the mixture was concentrated under reduced pressure. The obtained residue was purified by silica gel column chromatography (chloroform / methanol = 100/0) to obtain (9Z, 9'Z, 12Z, 12'Z) -2- (dimethylamino) -2-(((9Z, 12Z) -Octadeca-9,12-dienoyloxy) methyl) propane-1,3-diyl dioctadeca-9,12-dienoate (0.334 g, 0.356 mmol, 21% yield) was obtained.
ESI-MS m / z: 937 (M + H) ⁺ .
Process 2
(9Z, 9'Z, 12Z, 12'Z) -2- (dimethylamino) -2-(((9Z, 12Z) -octadec-9,12-dienoyloxy) methyl) propane-1 obtained in step 1 Methyl iodide (0.216 mL) was added to a chloroform (3 mL) solution of 1,3-diyl dioctadec-9,12-dienoate (0.324 g, 0.346 mmol), and the mixture was stirred at room temperature for 5 hours. Methyl iodide (0.216 mL) was added to the reaction mixture, and the mixture was stirred at room temperature overnight. The reaction solution was concentrated under reduced pressure. Dissolve the residue in a small amount of methanol-chloroform (1: 1) to ion exchange resin (Dow Chemical, Dowex (TM) 1x-2 100 mesh, Cl type, approx. Loaded and eluted with methanol-chloroform (1: 1). The eluate was concentrated under reduced pressure, and the resulting residue was purified by silica gel column chromatography (chloroform / methanol = 100/0 to 90/10) to give the title compound (0.161 g, 0.164 mmol, 47% yield). Obtained.
ESI-MS m / z: 951 (M) ⁺ ; ¹ H-NMR (CDCl ₃ ) δ: 0.86 (t, J = 6.9 Hz, 9H), 1.22-1.36 (m, 42H), 1.54-1.61 (m, 6H), 2.02 (q, J = 6.8 Hz, 12H), 2.34 (t, J = 7.7 Hz, 6H), 2.74 (t, J = 6.8 Hz, 6H), 3.69 (s, 9H), 4.57 (s, 6H), 5.26-5.39 (m, 12H).

N, N, N-trimethyl-1,3-bis ((Z) -tetradec-9-enoyloxy) -2-(((Z) -tetradec-9-enoyloxy) methyl) propane-2-aminium chloride (Compound II -2)
In the same manner as in Example 8, using cis-9-tetradecenoic acid (Nu-Chek Prep, Inc.) instead of (9Z, 12Z) -octadeca-9,12-dienoic acid, the title compound (0.0854 g, 0.104 mmol, through yield 16%).
ESI-MS m / z: 789 (M) ⁺ ; ¹ H-NMR (CDCl ₃ ) δ: 0.89 (t, J = 7.1 Hz, 9H), 1.27-1.36 (m, 36H), 1.58-1.64 (m, 6H), 2.02 (q, J = 6.5 Hz, 12H), 2.37 (t, J = 7.6 Hz, 6H), 3.72 (s, 9H), 4.55 (s, 6H), 5.30-5.38 (m, 6H).

N, N, N-trimethyl-1,3-bis (oleoyloxy) -2- (oleoyloxymethyl) propane-2-aminium chloride (Compound II-3)
In the same manner as in Example 8, using oleic acid instead of (9Z, 12Z) -octadeca-9,12-dienoic acid, the title compound (1.14 g, 1.15 mmol, overall yield 34%) was obtained. .
ESI-MS m / z: 957 (M) ⁺ ; ¹ H-NMR (CDCl ₃ ) δ: 0.83 (t, J = 6.9 Hz, 9H), 1.17-1.32 (m, 60H), 1.51-1.59 (m, 6H), 1.96 (t, J = 5.5 Hz, 12H), 2.32 (t, J = 7.6 Hz, 6H), 3.70 (s, 9H), 4.56 (s, 6H), 5.25-5.34 (m, 6H).

N, N, N-Trimethyl-1,3-bis (stearoyloxy) -2- (stearoyloxymethyl) propane-2-aminium chloride (Compound II-4)
Process 1
2- (dimethylamino) -2- (hydroxymethyl) propane-1,3-diol (0.100 g, 0.670 mmol) to toluene (10 mL), stearic acid (manufactured by Tokyo Chemical Industry Co., Ltd., 0.763 g, 2.68 mmol), p-Toluenesulfonic acid monohydrate (0.191 g, 1.01 mmol) was added in order, and the mixture was stirred overnight under heating under reflux. The reaction mixture was concentrated under reduced pressure, and the resulting residue was purified by amino silica gel column chromatography (chloroform) to give 2- (dimethylamino) -2-((stearoyloxy) methyl) propane-1,3-diyl distearate. (0.120 g, 0.127 mmol, 19% yield) was obtained.
ESI-MS m / z: 948 (M + H) ⁺
Process 2
Using 2- (dimethylamino) -2-((stearoyloxy) methyl) propane-1,3-diyl distearate (0.120 g, 0.127 mmol) obtained in Step 1, as in Step 1 of Example 1. The title compound (0.0260 g, 0.0260 mmol, yield 21%) was obtained.
ESI-MS m / z: 963 (M) ⁺ ; ¹ H-NMR (CDCl ₃ ) δ: 0.88 (t, J = 7.0 Hz, 9H), 1.22-1.36 (m, 84H), 1.56-1.65 (m, 6H), 2.37 (t, J = 7.6 Hz, 6H), 3.72 (s, 9H), 4.56 (s, 6H).

1,3-bis ((Z) -hexadec-9-enoyloxy) -2-(((Z) -hexadec-9-enoyloxy) methyl) -N, N, N-trimethylpropane-2-aminium chloride (compound II -Five)
In the same manner as in Example 8, using cis-9-hexadecenoic acid instead of (9Z, 12Z) -octadeca-9,12-dienoic acid, the title compound (0.680 g, 0.748 mmol, overall yield 63% )
ESI-MS m / z: 873 (M) ⁺ ; ¹ H-NMR (CDCl ₃ ) δ: 0.88 (t, J = 7.0 Hz, 9H), 1.24-1.36 (m, 48H), 1.56-1.67 (m, 13H), 1.98-2.05 (m, 12H), 2.37 (t, J = 7.6 Hz, 6H), 3.75 (s, 9H), 4.53 (s, 6H), 5.29-5.40 (m, 6H).

N, N, N-trimethyl-1,3-bis ((9Z, 12Z) -octadeca-9,12-dienyloxy) -2-(((9Z, 12Z) -octadeca-9,12-dienyloxy) methyl) propane -2-Aminium chloride (Compound II-6)
Process 1
Sodium hydride (Nacalai Tesque, oily, 60%) in a solution of 2-dimethylamino-2-hydroxymethylpropane-1,3-diol (Zylexa Pharma, 0.115 g, 0.768 mmol) in toluene (5 mL) 0.154 g, 3.84 mmol) and (9Z, 12Z) -octadeca-9,12-dienyl methanesulfonate (1.32 g, 3.84 mmol) were added, and the mixture was stirred overnight with heating under reflux. After cooling to room temperature, saturated aqueous sodium hydrogen carbonate solution was added to the reaction mixture, and the mixture was extracted with hexane. The organic layer was dried over anhydrous magnesium sulfate, filtered and concentrated under reduced pressure. The resulting residue was purified by amino silica gel column chromatography (hexane / ethyl acetate = 100/0 to 95/5) to give N, N-dimethyl-1,3-bis ((9Z, 12Z) -octadeca-9, 12-dienyloxy) -2-(((9Z, 12Z) -octadeca-9,12-dienyloxy) methyl) propan-2-amine (0.195 g, 0.217 mmol, 28% yield) was obtained.
ESI-MS m / z: 895 (M + H) ⁺ .
Process 2
N, N-dimethyl-1,3-bis ((9Z, 12Z) -octadeca-9,12-dienyloxy) -2-(((9Z, 12Z) -octadeca-9,12-dienyloxy) obtained in step 1 Methyl iodide (0.119 mL) was added to a solution of) methyl) propan-2-amine (0.0849 g, 0.0949 mmol) in chloroform (1 mL), and the mixture was stirred at room temperature overnight. The reaction solution was concentrated under reduced pressure. Dissolve the residue in a small amount of methanol-chloroform (1: 1) to ion exchange resin (Dow Chemical, Dowex (TM) 1x-2 100 mesh, Cl type, approx. Loaded and eluted with methanol-chloroform (1: 1). The eluate was concentrated under reduced pressure, and the resulting residue was purified by silica gel column chromatography (chloroform / methanol = 100/0 to 90/10) to give the title compound (0.0646 g, 0.0684 mmol, 72% yield). Obtained.
ESI-MS m / z: 909 (M) ⁺ ; ¹ H-NMR (CDCl ₃ ) δ: 0.89 (t, J = 6.9 Hz, 9H), 1.25-1.40 (m, 48H), 1.55-1.63 (m, 6H), 2.02-2.09 (m, 12H), 2.77 (t, J = 6.8 Hz, 6H), 3.44 (t, J = 6.6 Hz, 6H), 3.62 (s, 9H), 3.82 (s, 6H), 5.29-5.42 (m, 12H).

N, N, N-trimethyl-1,3-bis ((Z) -tetradec-9-enyloxy) -2-(((Z) -tetradec-9-enyloxy) methyl) propane-2-aminium chloride (compound II -7)
In the same manner as in Example 13, using myristoyl methanesulfonate (manufactured by Nu-Chek Prep, Inc) instead of (9Z, 12Z) -octadeca-9,12-dienyl methanesulfonate, the title compound (0.0729 g, 0.0931 mmol, through yield 12%).
ESI-MS m / z: 747 (M) ⁺ ; ¹ H-NMR (CDCl ₃ ) δ: 0.90 (t, J = 7.1 Hz, 9H), 1.27-1.37 (m, 42H), 1.54-1.61 (m, 6H), 2.02 (q, J = 6.5 Hz, 12H), 3.43 (t, J = 6.6 Hz, 6H), 3.64 (s, 8H), 3.81 (s, 6H), 5.31-5.39 (m, 6H).

1,3-bis ((Z) -hexadec-9-enyloxy) -2-(((Z) -hexadeca-9-enyloxy) methyl) -N, N, N-trimethylpropane-2-aminium chloride (compound II -8)
In the same manner as in Example 13, instead of (9Z, 12Z) -octadeca-9,12-dienyl methanesulfonate, palmitoleyl methanesulfonate (manufactured by Nu-Chek Prep, Inc.) was used, and g, 0.538 mmol, yield 71%).
ESI-MS m / z: 831 (M) ⁺ ; ¹ H-NMR (CDCl ₃ ) δ: 0.88 (t, J = 6.8 Hz, 9H), 1.23-1.37 (m, 54H), 1.53-1.61 (m, 14H), 2.02 (q, J = 5.8 Hz, 12H), 3.43 (t, J = 6.5 Hz, 6H), 3.65 (s, 9H), 3.81 (s, 6H), 5.30-5.40 (m, 6H).

(6Z, 9Z, 40Z, 43Z) -N, N, N-trimethyl-25-((3-((9Z, 12Z) -octadeca-9,12-dienyloxy) -3-oxopropoxy) methyl) -20, 30-dioxo-19,23,27,31-tetraoxanonatetraconta-6,9,40,43-tetraene-25-aminium chloride (compound II-9)
Process 1
Di-tert-butyl 3,3 ′ synthesized by a method according to the method described in “Journal of Organic Chemistry”, 2002, Vol. 67, p.1411-1413 -((2-amino-2-((3- (tert-butoxy) -3-oxopropoxy) methyl) propane-1,3-diyl) bis (oxy)) dipropanoate (0.500 g, 0.989 mmol) in dichloromethane ( 5 mL), methyl iodide (1.40 g, 9.89 mmol) was added, and the mixture was stirred at room temperature overnight. The reaction solution was concentrated under reduced pressure. Dissolve the residue in a small amount of methanol-chloroform (1: 1) and load it onto an ion exchange resin (Dowex (TM) 1x-2 100 mesh, Cl type, approx. 20 times volume, pre-washed with water and methanol). Eluted with. The eluate was concentrated under reduced pressure, and the resulting residue was purified by silica gel column chromatography (chloroform / methanol = 97 / 3-80 / 20) to give 9-((3- (tert-butoxy) -3-oxopropoxy. ) Methyl) -N, N, N, 2,2,16,16-heptamethyl-4,14-dioxo-3,7,11,15-tetraoxaheptadecan-9-aminium chloride (0.144 g, 0.246 mmol, Yield 25%).
ESI-MS m / z: 548 (M + H) ⁺ .
Process 2
9-((3- (tert-butoxy) -3-oxopropoxy) methyl) -N, N, N, 2,2,16,16-heptamethyl-4,14-dioxo-3 obtained in step 1 Dissolve 7,11,15-tetraoxaheptadecane-9-aminium chloride (0.350 g, 0.246 mmol) in dichloromethane (2 mL), add trifluoroacetic acid (0.380 mL, 4.92 mmol), and at room temperature for 3 hours. Stir. Toluene was added to the reaction solution and concentrated under reduced pressure to obtain 1,3-bis (2-carboxyethoxy) -2-((2-carboxyethoxy) methyl) -N, N, N-trimethylpropane-2- A crude product of aminium chloride trifluoroacetate (0.102 g, 0.246 mmol, crude yield 100%) was obtained.
ESI-MS m / z: 422 (M + H) ⁺
Process 3
Crude formation of 1,3-bis (2-carboxyethoxy) -2-((2-carboxyethoxy) methyl) -N, N, N-trimethylpropane-2-aminium chloride trifluoroacetate obtained in step 2 (0.055 g, 0.13 mmol) was dissolved in dichloromethane (2 mL) and O- (7-aza-1H-benzotriazol-1-yl) -N, N, N ', N'-tetramethyluronium hexa Fluorophosphate (Wako Pure Chemical Industries, 0.20 g, 0.53 mmol), N, N-diisopropylethylamine (0.23 mL, 1.3 mmol), and (9Z, 12Z) -octadeca-9,12-dien-1-ol (Tokyo Chemical Industry Co., Ltd., 0.141 g, 0.53 mmol) was added, and the mixture was stirred overnight at room temperature. Water was added to the reaction solution and extracted with chloroform. The organic layer was washed with saturated brine, dried over anhydrous magnesium sulfate, filtered, and concentrated under reduced pressure. The obtained residue was purified by silica gel column chromatography (chloroform / methanol = 90 / 10-80 / 20) to obtain the title compound (8.0 mg, 6.9 mmol, yield 5%).
ESI-MS m / z: 1125 (MH) ⁺ ; ¹ H-NMR (CDCl ₃ ) δ: 0.89 (t, J = 7.0 Hz, 9H), 1.25-1.39 (m, 48H), 1.58-1.66 (m, 6H), 2.05 (q, J = 6.9 Hz, 12H), 2.59 (t, J = 5.7 Hz, 6H), 2.77 (t, J = 6.7 Hz, 6H), 3.42 (s, 9H), 3.74 (t, J = 5.7 Hz, 6H), 4.00 (s, 6H), 4.07 (t, J = 6.8 Hz, 6H), 5.29-5.40 (m, 12H).

(7Z, 38Z) -23-((3-((Z) -Hexadeca-9-enyloxy) -3-oxopropoxy) methyl) -N, N, N-trimethyl-18,28-dioxo-17,21, 25,29-Tetraoxapentatetraconta-7,38-diene-23-aminium chloride (Compound II-10)
In the same manner as in Example 16, instead of (9Z, 12Z) -octadeca-9,12-dien-1-ol, (Z) -hexadeca-9-en-1-ol (Nu-Chek Prep, Inc. The title compound (0.145 g, 0.134 mmol, overall yield 17%) was obtained.
ESI-MS m / z: 1047 (M) ⁺ ; ¹ H-NMR (CDCl ₃ ) δ: 0.88 (t, J = 7.0 Hz, 9H), 1.24-1.38 (m, 54H), 1.58-1.66 (m, 6H), 1.98-2.05 (m, 12H), 2.58 (t, J = 5.7 Hz, 6H), 3.47 (s, 9H), 3.74 (t, J = 5.7 Hz, 6H), 4.02 (s, 6H), 4.07 (t, J = 6.8 Hz, 6H), 5.30-5.40 (m, 6H).

(5Z, 36Z) -N, N, N-trimethyl-16,26-dioxo-21-((3-oxo-3-((Z) -tetradec-9-enyloxy) propoxy) methyl) -15,19, 23,27-Tetraoxachentetraconta-5,36-diene-21-aminium chloride (Compound II-11)
In the same manner as in Example 16, instead of (9Z, 12Z) -octadeca-9,12-dien-1-ol, (Z) -tetradec-9-en-1-ol (Nu-Chek Prep, Inc. The title compound (0.189 g, 0.189 mmol, overall yield 24%) was obtained.
ESI-MS m / z: 963 (M) ⁺ ; ¹ H-NMR (CDCl ₃ ) δ: 0.87-0.92 (m, 9H), 1.25-1.38 (m, 42H), 1.55-1.66 (m, 6H), 1.98-2.05 (m, 12H), 2.58 (t, J = 5.7 Hz, 6H), 3.47 (s, 9H), 3.75 (t, J = 5.7 Hz, 6H), 4.01 (s, 6H), 4.07 (t , J = 6.8 Hz, 6H), 5.30-5.41 (m, 6H).

(11Z, 14Z) -N, N, N-trimethyl-2-(((9Z, 12Z) -octadeca-9,12-dienoyloxy) methyl) -2-((9Z, 12Z) -octadeca-9,12- Dienyl) icosa-11,14-dien-1-aminium chloride (compound II-12)
Process 1
Dissolve ethyl cyanoacetate (Tokyo Chemical Industry, 1.00 g, 8,84 mmol) and (9Z, 12Z) -octadeca-9,12-dienyl methanesulfonate (7.62 g, 22.1 mmol) in tetrahydrofuran (30 mL) Under ice-cooling, sodium hydride (oil, 60%, 1.06 g, 26.5 mmol) and tetra-n-butylammonium iodide (Nacalai Tesque, 3.27 g, 8.84 mmol) were added. After foaming subsided, the mixture was stirred at 60 ° C. for 3 hours. Water was added to the reaction solution and extracted with hexane. The organic layer was washed with saturated brine, dried over anhydrous magnesium sulfate, and filtered. By concentrating under reduced pressure, a crude product of (11Z, 14Z) -ethyl 2-cyano-2-((9Z, 12Z) -octadeca-9,12-dien-1-yl) icosa-11,14-dienoate (3.50 g, 5.74 mmol, crude yield 65%) was obtained.
Process 2
Crude product of (11Z, 14Z) -ethyl 2-cyano-2-((9Z, 12Z) -octadeca-9,12-dien-1-yl) icosa-11,14-dienoate obtained in Step 1 1.50 g, 2.46 mmol) was dissolved in tetrahydrofuran (10 mL), and lithium aluminum hydride (manufactured by Junsei Chemical Co., Ltd., 0.467 g, 12.3 mmol) was added under ice cooling, followed by stirring for 30 minutes. Water (0.5 mL), 15% aqueous sodium hydroxide solution (0.5 mL), water (1.5 mL) and magnesium sulfate were sequentially added to the reaction mixture, and the mixture was stirred for a while and then filtered. The residue obtained by concentration under reduced pressure was purified by silica gel column chromatography (chloroform / methanol = 99/1 to 85/15) to obtain (11Z, 14Z) -2- (aminoethyl) -2-((9Z, 12Z) -octadeca-9,12-dien-1-yl) icosa-11,14-dien-1-ol (1.00 g, 2.46 mmol, 71% yield) was obtained.
ESI-MS m / z: 573 (M + H) ⁺ .
Process 3
(11Z, 14Z) -2- (Aminoethyl) -2-((9Z, 12Z) -octadeca-9,12-dien-1-yl) icosa-11,14-diene-1- obtained in Step 2 All (0.350 g, 0.612 mmol) dissolved in acetonitrile (2 mL) and tetrahydrofuran (2 mL), 38% aqueous formaldehyde solution (Wako Pure Chemical Industries, 0.145 mL, 1.84 mmol), acetic acid (0.035 mL, 0.612 mmol) ) And sodium triacetoxyborohydride (Acros Organics, 0.389 g, 1.84 mmol) were added, and the mixture was stirred at room temperature overnight. Water was added to the reaction solution and extracted with chloroform. The organic layer was washed with saturated brine, dried over anhydrous magnesium sulfate, and filtered. The obtained residue was purified by silica gel column chromatography (chloroform / methanol = 99/1 to 85/15) to obtain (11Z, 14Z) -2-((dimethylamino) methyl) -2-((9Z, 12Z) -Octadeca-9,12-dien-1-yl) icosa-11,14-dien-1-ol (0.252 g, 0.420 mmol, yield 69%) was obtained.
ESI-MS m / z: 600 (M + H) ⁺
Process 4
(11Z, 14Z) -2-((Dimethylamino) methyl) -2-((9Z, 12Z) -octadeca-9,12-dien-1-yl) icosa-11,14-diene obtained in Step 3 To a solution of 1-ol (0.252 g, 0.420 mmol) in dichloromethane (4 mL), (9Z, 12Z) -octadeca-9,12-dienoic acid (0.141 g, 0.504 mmol), O- (7-aza-1H -Benzotriazol-1-yl) -N, N, N ', N'-tetramethyluronium hexafluorophosphate (0.192 mmol, 0.504 mmol), N, N-diisopropylethylamine (0.147 mL, 0.840 mmol) in this order In addition, the mixture was stirred at room temperature for 4 hours. Water was added to the reaction solution and the mixture was extracted with hexane. The organic layer was washed with water, dried over anhydrous magnesium sulfate, filtered and concentrated under reduced pressure. The obtained residue was purified by silica gel column chromatography (hexane / ethyl acetate = 95/5 to 85/15) to obtain (9Z, 12Z)-(11Z, 14Z) -2-((dimethylamino) methyl) -2-((9Z, 12Z) -octadeca-9,12-dien-1-yl) icosa-11,14-dien-1-yl octadec-9,12-dienoate (0.307 g, 0.356 mmol, yield 85 %).
ESI-MS m / z: 863 (M + H) ⁺
Process 5
(9Z, 12Z)-(11Z, 14Z) -2-((Dimethylamino) methyl) -2-((9Z, 12Z) -octadeca-9,12-dien-1-yl) icosa obtained in step 4 -11,14-Dien-1-yl octadeca-9,12-dienoate (0.307 g, 0.356 mmol) was used in the same manner as in Example 1, step 2 to give the title compound (0.260 g, 0.285 mmol, 80% yield). )
ESI-MS m / z: 877 (M) ⁺ ; ¹ H-NMR (CDCl ₃ ) δ: 0.89 (t, J = 7.0 Hz, 9H), 1.22-1.48 (m, 54H), 1.60-1.66 (m, 2H), 2.05 (q, J = 6.8 Hz, 12H), 2.38 (t, J = 7.6 Hz, 2H), 2.77 (t, J = 6.3 Hz, 6H), 3.50 (s, 2H), 3.60 (s, 9H), 4.13 (s, 2H), 5.27-5.44 (m, 12H).

N, N, N-trimethyl-3-((11Z, 14Z) -2-(((9Z, 12Z) -octadeca-9,12-dienoyloxy) methyl) -2-((9Z, 12Z) -octadeca-9 , 12-Dienyl) icosa-11,14-dienylcarbamoyloxy) propane-1-aminium chloride (Compound II-13)
Process 1
Example 19 (11Z, 14Z) -2- (Aminoethyl) -2-((9Z, 12Z) -octadeca-9,12-dien-1-yl) icosa-11,14-diene obtained in Step 2 -1-ol (0.918 g, 1.61 mmol) is dissolved in tetrahydrofuran (20 mL), triethylamine (0.671 mL, 4.81 mmol), and di-tert-butyl dicarbonate (manufactured by Kokusan Chemical Co., Ltd., 0.373 mL, 1.61 mmol) And stirred at room temperature overnight. The reaction mixture was concentrated under reduced pressure, and the obtained residue was purified by silica gel column chromatography (hexane / ethyl acetate = 95 / 5-50 / 50) to give tert-butyl ((11Z, 14Z) -2- ( Hydroxymethyl) -2-((9Z, 12Z) -octadeca-9,12-dien-1-yl) icosa-11,14-dien-1-yl) carbamate (0.918 g, 1.37 mmol, 85% yield) Got.
ESI-MS m / z: 672 (M + H) ⁺ .
Process 2
Tert-butyl ((11Z, 14Z) -2- (hydroxymethyl) -2-((9Z, 12Z) -octadeca-9,12-dien-1-yl) icosa-11,14- Dien-1-yl) carbamate (0.357 g, 0.531 mmol) was dissolved in dichloromethane (5 mL) and (9Z, 12Z) -octadeca-9,12-dienoic acid (0.223 g, 0.797 mmol), O- (7 -Aza-1H-benzotriazol-1-yl) -N, N, N ', N'-tetramethyluronium hexafluorophosphate (0.303 mmol, 0.797 mmol), N, N-diisopropylethylamine (0.186 mL, 1.06 mmol) and N, N-dimethylaminopyridine (0.0650 g, 0.531 mmol) were added, and the mixture was stirred overnight at room temperature. Water was added to the reaction solution and extracted with chloroform. The organic layer was washed with saturated brine, dried over anhydrous magnesium sulfate, filtered, and concentrated under reduced pressure. The obtained residue was purified by silica gel column chromatography (hexane / ethyl acetate = 99 / 1-90 / 10) to give (9Z, 12Z)-(11Z, 14Z) -2-(((tert-butoxycarbonyl Amino) methyl) -2-((9Z, 12Z) -octadeca-9,12-dien-1-yl) icosa-11,14-dien-1-yl octadec-9,12-dienoate (0.395 g, 0.423 mmol Yield 80%).
ESI-MS m / z: 935 (M + H) ⁺ .
Process 3
(9Z, 12Z)-(11Z, 14Z) -2-(((tert-butoxycarbonylamino) methyl) -2-((9Z, 12Z) -octadeca-9,12-diene-1 obtained in Step 2 -Il) icosa-11,14-dien-1-yl octadeca-9,12-dienoate (0.395 g, 0.423 mmol) was dissolved in dichloromethane (3 mL) and trifluoroacetic acid (1.00 mL, 4.92 mmol) under ice-cooling. ) Was added and stirred for 2 hours at 0 ° C. 1,2-dichloroethane was added to the reaction solution and concentrated under reduced pressure to obtain (9Z, 12Z)-(11Z, 14Z) -2- (aminoethyl) -2-((9Z, 12Z) -octadeca-9,12-dien-1-yl) icosa-11,14-dien-1-yl octadec-9,12-dienoate trifluoroacetate crude product (0.394 g, 0.423 mmol, crude yield 100%).
ESI-MS m / z: 834 (M + H) ⁺ .
Process 4
(9Z, 12Z)-(11Z, 14Z) -2- (aminoethyl) -2-((9Z, 12Z) -octadeca-9,12-dien-1-yl) icosa-11 obtained in Step 3 14-Dien-1-yl octadeca-9,12-dienoate Trifluoroacetate crude product (0.200 g, 0.215 mmol) was dissolved in acetonitrile (2 mL) and “Journal of American Chemical Society ( J. Am. Chem. Soc.), 1981, Vol. 103, p. 4194-4199, synthesized by a method according to the method described in 3- (dimethylamino) propyl 4-nitrophenyl carbonate hydrochloride (0.279 g 1.07 mmol), triethylamine (0.299 mL, 2.15 mmol), and N, N-dimethylaminopyridine (0.0520 g, 0.429 mmol) were added, and the mixture was stirred at 60 ° C. for 2 hours. Saturated aqueous sodium hydrogen carbonate solution was added to the reaction mixture, and the mixture was extracted with chloroform. The organic layer was washed with saturated brine, dried over anhydrous magnesium sulfate, filtered, and concentrated under reduced pressure. The obtained residue was purified by silica gel column chromatography (hexane / ethyl acetate = 99 / 1-60 / 40) to obtain (9Z, 12Z)-(11Z, 14Z) -2-((((3- ( Dimethylamino) propoxy) carbonyl) amino) methyl) -2-((9Z, 12Z) -octadeca-9,12-dien-1-yl) icosa-11,14-dien-1-yl octadec-9,12- Dienoate (0.0800 g, 0.0830 mmol, 39% yield) was obtained.
ESI-MS m / z: 964 (M + H) ⁺ .
Process 5
(9Z, 12Z)-(11Z, 14Z) -2-((((3- (Dimethylamino) propoxy) carbonyl) amino) methyl) -2-((9Z, 12Z) -octadeca- 9,12-Dien-1-yl) icosa-11,14-dien-1-yloctadeca-9,12-dienoate (0.053 g, 0.055 mmol) was used in the same manner as in Example 1, Step 2 to give the title. The compound (0.025 g, 0.025 mmol, yield 45%) was obtained.
ESI-MS m / z: 978 (M) ⁺ ; ¹ H-NMR (CDCl ₃ ) δ: 0.89 (t, J = 7.0 Hz, 9H), 1.17-1.40 (m, 54H), 1.56-1.66 (m, 2H), 2.05 (q, J = 6.8 Hz, 12H), 2.09-2.17 (m, 2H), 2.33 (t, J = 7.6 Hz, 2H), 2.77 (t, J = 6.2 Hz, 6H), 3.05 ( d, J = 6.6 Hz, 2H), 3.44 (s, 9H), 3.73-3.79 (m, 2H), 3.85 (s, 2H), 4.16 (t, J = 5.7 Hz, 2H), 5.27-5.44 (m , 12H), 5.72 (t, J = 6.5 Hz, 1H).

(12Z, 15Z) -3-Hydroxy-N, N, N-trimethyl-2,2-di ((9Z, 12Z) -octadeca-9,12-dien-1-yl) henikosa-12,15-diene- 1-Aminium chloride (Compound II-14)
Process 1
Example 19 (11Z, 14Z) -2- (Aminoethyl) -2-((9Z, 12Z) -octadeca-9,12-dien-1-yl) icosa-11,14-diene obtained in Step 2 1-ol (1.35 g, 2.36 mmol) dissolved in tetrahydrofuran (10 mL), 38% formaldehyde aqueous solution (0.559 mL, 7.08 mmol, Wako Pure Chemical Industries, Ltd.), acetic acid (0.135 mL, 2.36 mmol), and sodium Triacetoxyborohydride (1.50 g, 7.08 mmol) was added, and the mixture was stirred at room temperature for 1 hour. Water was added to the reaction mixture, and the mixture was extracted with chloroform. The organic layer was washed with saturated brine, dried over anhydrous magnesium sulfate, filtered, and concentrated under reduced pressure. The obtained residue was purified by silica gel column chromatography (chloroform / methanol = 99/1 to 85/15) to give (11Z, 14Z) -2-((dimethylamino) methyl) -2-((9Z, 12Z) -octadeca-9,12-dien-1-yl) icosa-11,14-dien-1-ol (0.610 g, 1.02 mmol, 43% yield) was obtained.
ESI-MS m / z: 600 (M + H) ⁺ .
Process 2
(11Z, 14Z) -2-((Dimethylamino) methyl) -2-((9Z, 12Z) -octadeca-9,12-dien-1-yl) icosa-11,14-diene obtained in Step 1 -1-ol (0.300 g, 0.500 mmol) was dissolved in dichloromethane (3 mL), Dess-Martin reagent (manufactured by Tokyo Chemical Industry Co., Ltd., 0.233 g, 0.550 mmol) was added, and the mixture was stirred at room temperature for 1 hour. Water was added to the reaction mixture, and the mixture was extracted with chloroform. The organic layer was washed with saturated brine, dried over anhydrous magnesium sulfate, filtered, and concentrated under reduced pressure. The obtained residue was purified by silica gel column chromatography (hexane / ethyl acetate = 95/5 to 70/30) to give (11Z, 14Z) -2-((dimethylamino) methyl) -2-((9Z , 12Z) -octadeca-9,12-dien-1-yl) icosa-11,14-dienal (0.160 g, 0.268 mmol, 54% yield) was obtained.
ESI-MS m / z: 598 (M + H) ⁺ .
Process 3
Diethyl ether (1 mL) and iodine (one piece) were added to magnesium (Sigma-Ardrich, 0.0140 g, 0.562 mmol), and the mixture was stirred at room temperature for 5 minutes. There, a diethyl ether solution of (6Z, 9Z) -18-bromooctadeca-6,9-diene (0.176 g, 0.535 mmol) synthesized by a method according to the method described in International Publication No. 2010/42877 ( 1 mL) was added, and the mixture was stirred with heating under reflux. After confirming that the color of iodine disappeared, (11Z, 14Z) -2-((dimethylamino) methyl) -2-((9Z, 12Z) -octadeca-9,12-diene obtained in Step 2 was obtained. A solution of 1-yl) icosa-11,14-dienal (0.160 g, 0.268 mmol) in diethyl ether (1 mL) was added, and the mixture was stirred at room temperature for 1 hour. Water was added to the reaction mixture, and the mixture was extracted with hexane. The organic layer was washed with saturated brine, dried over anhydrous magnesium sulfate, filtered, and concentrated under reduced pressure. The obtained residue was purified by silica gel column chromatography (hexane / ethyl acetate = 99/1 to 85/15) to give (6Z, 9Z, 29Z, 32Z) -20-((dimethylamino) methyl) -20 -((9Z, 12Z) -octadeca-9,12-dien-1-yl) octatriconta-6,9,29,32-tetraen-19-ol (0.0470 g, 0.0550 mmol, 21% yield) Obtained.
ESI-MS m / z: 848 (M + H) ⁺ .
Process 4
(6Z, 9Z, 29Z, 32Z) -20-((Dimethylamino) methyl) -20-((9Z, 12Z) -octadeca-9,12-dien-1-yl) octatria contour obtained in Step 3 -6,9,29,32-tetraen-19-ol (0.047 g, 0.055 mmol) was used in the same manner as in Example 1, step 2 to give the title compound (0.0012 g, 0.0013 mmol, yield 2%). Obtained.
ESI-MS m / z: 863 (M) ⁺ ; ¹ H-NMR (CDCl ₃ ) δ: 0.89 (t, J = 7.0 Hz, 9H), 1.17-1.40 (m, 58H), 1.54-1.65 (m, 2H), 2.05 (q, J = 6.8 Hz, 12H), 2.77 (t, J = 6.5 Hz, 6H), 3.29 (d, J = 14.4 Hz, 1H), 3.51 (s, 9H), 3.56 (d, J = 14.2 Hz, 1H), 3.62-3.70 (m, 1H), 5.29-5.42 (m, 12H).

(11Z, 14Z) -N, N, N-trimethyl-2-((9Z, 12Z) -octadeca-9,12-dien-1-yl) -2-(((9Z, 12Z) -octadeca-9, 12-Dienyloxy) carbonyl) icosa-11,14-dien-1-aminium chloride (Compound II-15)
Process 1
Tert-Butyl ((11Z, 14Z) -2- (hydroxymethyl) -2-((9Z, 12Z) -octadeca-9,12-dien-1-yl) icosa- obtained in Step 1 of Example 20 11,14-Dien-1-yl) carbamate (0.300 g, 0.448 mmol) was dissolved in acetone (2 mL) and Jones reagent (Sigma-Ardrich, 2 mol / L, 0.224 mL, 0.448 mmol) was cooled with ice. ) And then stirred at room temperature for 1 hour. Water was added to the reaction mixture, and the mixture was extracted with ethyl acetate. The organic layer was washed with saturated brine, dried over anhydrous magnesium sulfate, filtered, and concentrated under reduced pressure. The obtained residue was purified by silica gel column chromatography (hexane / ethyl acetate = 95 / 5-50 / 50) to give (11Z, 14Z) -2-(((tert-butoxycarbonyl) amino) methyl)- 2-((9Z, 12Z) -octadeca-9,12-dien-1-yl) icosa-11,14-dienoic acid (0.136 g, 0.198 mmol, 44% yield) was obtained.
ESI-MS m / z: 684 (M-H) ^- .
Process 2
(11Z, 14Z) -2-(((tert-butoxycarbonyl) amino) methyl) -2-((9Z, 12Z) -octadeca-9,12-dien-1-yl) icosa- obtained in step 1 Using 11,14-dienoic acid (0.120 g, 0.175 mmol) and (9Z, 12Z) -octadeca-9,12-dien-1-ol (Nu-Chek Prep, Inc., 0.0930 g, 0.350 mmol) In the same manner as in Step 2 of Example 20, (11Z, 14Z)-(9Z, 12Z) -octadeca-9,12-dien-1-yl 2-((((tert-butoxycarbonyl) amino) methyl)- 2-((9Z, 12Z) -octadeca-9,12-dien-1-yl) icosa-11,14-dienoate (0.123 g, 0.132 mmol, 75% yield) was obtained.
ESI-MS m / z: 935 (M + H) ⁺ .
Process 3
(11Z, 14Z)-(9Z, 12Z) -Octadeca-9,12-dien-1-yl 2-((((tert-butoxycarbonyl) amino) methyl) -2-((9Z, (12Z) -octadeca-9,12-dien-1-yl) icosa-11,14-dienoate (0.123 g, 0.132 mmol) was dissolved in dichloromethane (1 mL) and trifluoroacetic acid (0.300 mL, 3.89 under ice-cooling). mmol) was added and stirred for 1 hour. Saturated aqueous sodium hydrogen carbonate solution was added to the reaction mixture, and the mixture was extracted with hexane. The organic layer was washed with saturated brine, dried over anhydrous magnesium sulfate, filtered, and concentrated under reduced pressure. The resulting residue was purified by silica gel column chromatography (hexane / ethyl acetate = 99 / 1-80 / 20) to give (11Z, 14Z)-(9Z, 12Z) -octadeca-9,12-diene-1 2-yl 2- (aminomethyl) -2-((9Z, 12Z) -octadeca-9,12-dien-1-yl) icosa-11,14-dienoate (0.110 g, 0.132 mmol, 100% yield) Obtained.
ESI-MS m / z: 835 (M + H) ⁺ .
Process 4
(11Z, 14Z)-(9Z, 12Z) -Octadeca-9,12-dien-1-yl 2- (aminomethyl) -2-((9Z, 12Z) -octadeca-9,12 obtained in Step 3 (11Z, 14Z)-(9Z, 12Z) -octadeca-9 in the same manner as in Example 21, step 1 using -dien-1-yl) icosa-11,14-dienoate (0.110 g, 0.132 mmol) , 12-Dien-1-yl 2-((dimethylaminomethyl) -2-((9Z, 12Z) -octadeca-9,12-dien-1-yl) icosa-11,14-dienoate (0.0720 g, 0.0830 mmol, yield 63%).
ESI-MS m / z: 862 (M + H) ⁺ .
Process 5
(11Z, 14Z)-(9Z, 12Z) -Octadeca-9,12-dien-1-yl 2-((dimethylaminomethyl) -2-((9Z, 12Z) -octadeca-9 obtained in Step 4 , 12-Dien-1-yl) icosa-11,14-dienoate (0.072 g, 0.083 mmol) in the same manner as in Example 1, Step 2, the title compound (0.052 g, 0.057 mmol, yield 68%). )
ESI-MS m / z: 877 (M) ⁺ ; ¹ H-NMR (CDCl ₃ ) δ: 0.89 (t, J = 7.0 Hz, 9H), 1.09-1.42 (m, 52H), 1.52-1.81 (m, 6H), 2.05 (q, J = 6.8 Hz, 12H), 2.77 (t, J = 6.6 Hz, 6H), 3.46 (s, 9H), 3.79 (s, 2H), 4.14 (t, J = 6.8 Hz, 2H), 5.28-5.43 (m, 12H).

(11Z, 14Z) -N, N, N-trimethyl-2,2-bis (((9Z, 12Z) -octadeca-9,12-dienoyloxy) methyl) icosa-11,14-dien-1-aminium chloride ( Compound II-16)
Process 1
Dimethyl malonate (Tokyo Chemical Industry Co., Ltd., 1.00 g, 7.57 mmol) was dissolved in acetonitrile (20 mL), (9Z, 12Z) -octadeca-9,12-dienyl methanesulfonate (2.61 g, 7.57 mmol), Cesium carbonate (Wako Pure Chemical Industries, 4.93 g, 15.1 mmol) and tetra-n-butylammonium iodide (2.80 g, 7.57 mmol) were added, and the mixture was stirred at 50 ° C. overnight. Water was added to the reaction mixture, and the mixture was extracted with hexane. The organic layer was washed with saturated brine, dried over anhydrous magnesium sulfate, filtered, and concentrated under reduced pressure. The resulting residue was purified by silica gel column chromatography (hexane / ethyl acetate = 90/10 to 70/30) to give dimethyl 2-((9Z, 12Z) -octadeca-9,12-dien-1-yl ) Malonate (1.22 g, 3.21 mmol, 42% yield) was obtained.
ESI-MS m / z: 381 (M + H) ⁺ .
Process 2
Dimethyl 2-((9Z, 12Z) -octadeca-9,12-dien-1-yl) malonate (0.200 g, 0.526 mmol) obtained in step 1 was dissolved in acetonitrile (3 mL), and N, N, N ′, N′-tetramethyldiaminomethane (manufactured by Tokyo Chemical Industry Co., Ltd., 0.0860 mL, 0.631 mmol) and acetic anhydride (0.0600 mL, 0.631 mmol) were added. Thereafter, sodium hydride (oil, 60%, 0.0320 g, 0.788 mmol) was added under ice cooling, and the mixture was stirred at room temperature for 3 hours. A saturated aqueous ammonium chloride solution was added to the reaction mixture, and the mixture was extracted with ethyl acetate. The organic layer was washed with saturated brine, dried over anhydrous magnesium sulfate, filtered, and concentrated under reduced pressure. The resulting residue was purified by silica gel column chromatography (hexane / ethyl acetate = 100/0 to 60/40) to give dimethyl 2-((dimethylamino) methyl) -2-((9Z, 12Z) -octadeca -9,12-dien-1-yl) malonate (0.0660 g, 0.151 mmol, 29% yield) was obtained.
ESI-MS m / z: 438 (M + H) ⁺ .
Process 3
Using the dimethyl 2-((dimethylamino) methyl) -2-((9Z, 12Z) -octadeca-9,12-dien-1-yl) malonate (0.066 g, 0.15 mmol) obtained in step 2, Example 19 In the same manner as in Step 2, 2-((dimethylamino) methyl) -2-((9Z, 12Z) -octadeca-9,12-dien-1-yl) propane-1,3-diol (0.013 g, 0.034 mmol, yield 23%) was obtained.
ESI-MS m / z: 382 (M + H) ⁺ .
Process 4
2-((Dimethylamino) methyl) -2-((9Z, 12Z) -octadeca-9,12-dien-1-yl) propane-1,3-diol obtained in Step 3 (0.013 g, 0.034 mmol ) And (9Z, 9'Z, 12Z, 12 ')-2-((dimethylamino) methyl) -2-((9Z, 12Z) -octadeca -9,12-dien-1-yl) propane-1,3-diylbis (octadeca-9,12-dienoate (0.017 g, 0.019 mmol, yield 56%) was obtained.
ESI-MS m / z: 906 (M + H) ⁺ .
Process 5
(9Z, 9′Z, 12Z, 12′Z) -2-((Dimethylamino) methyl) -2-((9Z, 12Z) -octadeca-9,12-dien-1-yl obtained in Step 4 ) Using propane-1,3-diylbis (octadeca-9,12-dienoate (0.017 g, 0.019 mmol) in the same manner as in Example 1, step 2, the title compound (5.5 mg, 0.0058 mmol, Rate 31%).
ESI-MS m / z: 921 (M) ⁺ . ¹ H-NMR (CDCl ₃ ) δ: 0.89 (t, J = 7.0 Hz, 9H), 1.23-1.40 (m, 48H), 1.53-1.65 (m, 4H), 2.05 (q, J = 6.9 Hz, 12H), 2.38 (t, J = 7.6 Hz, 4H), 2.77 (t, J = 6.6 Hz, 6H), 3.59 (s, 9H), 3.72 (s, 2H), 4.20 (dd, J = 22.1, 12.2 Hz, 4H), 5.28-5.44 (m, 12H).

N, N, N-trimethyl-3-((9Z, 12Z) -octadeca-9,12-dienoyloxy) -2,2-bis (((9Z, 12Z) -octadeca-9,12-dienoyloxy) methyl) propane -1-Aminium chloride (Compound II-17)
Process 1
2- (Bromomethyl) -2- (hydroxymethyl) propane-1,3-diol (0.200 g, 1.01 mmol) was mixed with dimethylamine (Sigma-Aldrich, 2.0 mol / L tetrahydrofuran solution, 5.02 mL, 10.1 mmol). In addition, the mixture was stirred at 120 ° C. for 15 hours under microwave irradiation. Lithium hydroxide monohydrate (0.0290 g, 1.21 mmol) was added to the reaction solution, and the resulting precipitate was filtered off. The filtrate was concentrated under reduced pressure to obtain a crude product of 2-((dimethylamino) methyl) -2- (hydroxymethyl) propane-1,3-diol (0.200 g, 1.23 mmol, quantitative). .
ESI-MS m / z: 164 (M + H) ⁺ .
Process 2
Using the crude product (0.200 g, 1.23 mmol) of 2-((dimethylamino) methyl) -2- (hydroxymethyl) propane-1,3-diol obtained in Step 1, the same method as in Example 8 Gave the title compound (0.0470 g, 0.047 mmol overall yield 4.4%).
ESI-MS m / z: 965 (M) ⁺ ; ¹ H-NMR (CDCl ₃ ) δ: 0.85-0.94 (m, 9H), 1.24-1.40 (m, 42H), 1.53-1.63 (m, 6H), 2.00-2.10 (m, 12H), 2.38 (t, J = 6.9 Hz, 6H), 2.77 (t, J = 6.5 Hz, 6H), 3.64 (s, 9H), 3.95 (s, 2H), 4.30 (s , 6H), 5.27-5.43 (m, 12H).

N, N, N-trimethyl-3- (oleoyloxy) -2,2-bis (oleoyloxymethyl) propane-1-aminium chloride (Compound II-18)
Using oleic acid in place of (9Z, 12Z) -octadeca-9,12-dienoic acid in Step 1 of Example 8, the title compound (0.663 g, 0.658 mmol, overall yield) was obtained in the same manner as in Example 8. 28%).
ESI-MS m / z: 971 (M) ⁺ ; ¹ H-NMR (CDCl ₃ ) δ: 0.88 (t, J = 6.8 Hz, 9H), 1.22-1.38 (m, 60H), 1.55-1.65 (m, 6H), 2.01 (q, J = 5.9 Hz, 12H), 2.38 (t, J = 7.6 Hz, 6H), 3.64 (s, 9H), 3.98 (s, 2H), 4.29 (s, 6H), 5.29- 5.39 (m, 6H).

N, N, N-trimethyl-3-((9Z, 12Z) -octadeca-9,12-dienyloxy) -2,2-bis (((9Z, 12Z) -octadeca-9,12-dienyloxy) methyl) propane -1-Aminium chloride (Compound II-19)
Process 1
Add 2- (bromomethyl) -2- (hydroxymethyl) propane-1,3-diol (1.52 g, 7.56 mmol) to dimethylamine (about 2 mol / L tetrahydrofuran solution, 15.0 mL, 30.0 mmol), and microwave reaction The mixture was stirred with heating at 120 ° C. for 15 hours. After cooling to room temperature, lithium hydroxide (0.217 g, 9.07 mmol) was added to the reaction mixture, filtered, and concentrated under reduced pressure to give 2-((dimethylamino) methyl) -2- (hydroxymethyl) propane-1, A crude product of 3-diol was obtained.
To the crude product obtained, toluene (30 mL), (9Z, 12Z) -octadeca-9,12-dienyl methanesulfonate (6.51 g, 18.9 mmol) and sodium hydride (oil, 60%, 0.756 g, 18.9 mmol) was added and stirred overnight under reflux with heating. After cooling to room temperature, saturated aqueous sodium hydrogen carbonate solution was added to the reaction mixture, and the mixture was extracted with hexane. The organic layer was dried over anhydrous magnesium sulfate, filtered and concentrated under reduced pressure. The obtained residue was purified by amino silica gel column chromatography (hexane / ethyl acetate = 100/0 to 90/10) to give N, N-dimethyl-3-((9Z, 12Z) -octadeca-9,12- Dienyloxy) -2,2-bis (((9Z, 12Z) -octadeca-9,12-dienyloxy) methyl) propan-1-amine (0.196 g, 0.216 mmol, 3%) and 3- (dimethylamino) -2 , 2-bis (((9Z, 12Z) -octadeca-9,12-dienyloxy) methyl) propan-1-ol (1.80 g, 2.73 mmol, yield 36%) was obtained.
N, N-dimethyl-3-((9Z, 12Z) -octadeca-9,12-dienyloxy) -2,2-bis (((9Z, 12Z) -octadeca-9,12-dienyloxy) methyl) propane-1 -Amine
ESI-MS m / z: 909 (M + H) ⁺ .
3- (Dimethylamino) -2,2-bis (((9Z, 12Z) -octadeca-9,12-dienyloxy) methyl) propan-1-ol
ESI-MS m / z: 661 (M + H) ⁺ .
Process 2
N, N-dimethyl-3-((9Z, 12Z) -octadeca-9,12-dienyloxy) -2,2-bis (((9Z, 12Z) -octadeca-9,12-dienyloxy) obtained in step 1 Methyl iodide (0.500 mL) was added to a solution of) methyl) propan-1-amine (0.120 g, 0.132 mmol) in chloroform (1 mL), and the mixture was stirred at room temperature for 3 hours. The reaction solution was concentrated under reduced pressure. Dissolve the residue in a small amount of methanol-chloroform (1: 1) and load it onto an ion exchange resin (Amberlite® IRA-400, Cl type, approximately 20 times volume, pre-washed with water and ethanol). Elute with (1: 1). The eluate was concentrated under reduced pressure, and the resulting residue was purified by silica gel column chromatography (chloroform / methanol = 100/0 to 90/10) to give the title compound (0.0654 g, 0.0682 mmol, 57% yield). Obtained.
ESI-MS m / z: 923 (M) ⁺ ; ¹ H-NMR (CDCl ₃ ) δ: 0.89 (t, J = 6.8 Hz, 9H), 1.22-1.40 (m, 1H), 1.49-1.59 (m, 6H), 2.05 (q, J = 6.9 Hz, 12H), 2.77 (t, J = 6.7 Hz, 6H), 3.37 (t, J = 6.6 Hz, 6H), 3.45 (s, 6H), 3.55 (s, 9H), 3.58 (s, 2H), 5.28-5.42 (m, 12H).

N, N, N-trimethyl-3-((9Z, 12Z) -octadeca-9,12-dienoyloxy) -2,2-bis (((9Z, 12Z) -octadeca-9,12-dienyloxy) methyl) propane -1-Aminium chloride (Compound II-20)
Example 26 3- (Dimethylamino) -2,2-bis (((9Z, 12Z) -octadeca-9,12-dienyloxy) methyl) propan-1-ol obtained in Step 1 (0.265 g, 0.401 mmol ) In 1,2-dichloroethane (4 mL) solution in (9Z, 12Z) -octadeca-9,12-dienoic acid (0.169 g, 0.602 mmol), 1-ethyl-3- (3-dimethylaminopropyl) carbodiimide hydrochloride Salt (0.154 g, 0.802 mmol) and N, N-dimethylaminopyridine (0.0250 g, 0.201 mmol) were added and stirred at room temperature overnight. Concentrated under reduced pressure. The obtained residue was purified by silica gel column chromatography (hexane / ethyl acetate = 95/5 to 90/10) to obtain (9Z, 12Z) -3- (dimethylamino) -2,2-bis (((9Z, A crude product of 12Z) -octadeca-9,12-dienyloxy) methyl) propyl octadeca-9,12-dienoate was obtained.
Chloroform (2 mL) and methyl iodide (manufactured by Tokyo Chemical Industry Co., Ltd., 1.00 mL) were added to the obtained crude product, and the mixture was stirred at room temperature for 5 hours. The reaction solution was concentrated under reduced pressure. Dissolve the residue in a small amount of methanol-chloroform (1: 1) and ion exchange resin (Sigma-Aldirch, Amberlite (R) IRA-400, Cl type, approximately 20 times volume, pre-washed with water and ethanol) Loaded and eluted with methanol-chloroform (1: 1). The eluate was concentrated under reduced pressure, and the resulting residue was purified by silica gel column chromatography (chloroform / methanol = 100/0 to 90/10) to give the title compound (0.220 g, 0.226 mmol, yield 56%). Obtained.
ESI-MS m / z: 937 (M) ⁺ ; ¹ H-NMR (CDCl ₃ ) δ: 0.89 (t, J = 7.0 Hz, 9H), 1.22-1.41 (m, 51H), 1.50-1.66 (m, 6H), 2.05 (q, J = 6.9 Hz, 12H), 2.38 (t, J = 7.5 Hz, 2H), 2.77 (t, J = 6.1 Hz, 6H), 3.39 (t, J = 6.6 Hz, 4H) , 3.44-3.48 (m, 2H), 3.54-3.58 (m, 11H), 3.73 (s, 2H), 4.18 (s, 2H), 5.28-5.43 (m, 11H).

N, N, N-trimethyl-4- (2- (9Z, 12Z) -octadeca-9,12-dienamido-3-((9Z, 12Z) -octadeca-9,12-dienoyloxy) -2-((( 9Z, 12Z) -Octadeca-9,12-dienoyloxy) methyl) propoxy) -4-oxybutane-1-aminium chloride (Compound II-21)
Process 1
To a solution of tert-butyl (1,3-dihydroxy-2- (hydroxymethyl) propan-2-yl) carbamate (Key Organics, 0.505 g, 2.28 mmol) in dichloromethane (15 mL) (9Z, 12Z) -Octadeca-9,12-dienoic acid (3.23 g, 11.4 mmol), 1-ethyl-3- (3-dimethylaminopropyl) carbodiimide hydrochloride (2.19 g, 11.4 mmol) and N, N-dimethylaminopyridine (0.279 g, 2.28 mmol) was added and stirred at room temperature for 1 hour. A saturated aqueous sodium hydrogen carbonate solution was added to the reaction solution, and the mixture was extracted with chloroform. The organic layer was dried over anhydrous magnesium sulfate, filtered and concentrated under reduced pressure. The resulting residue was purified by silica gel column chromatography (hexane / chloroform = 100/0 to 95/5) to give (9Z, 9'Z, 12Z, 12'Z) -2- (tert-butoxycarbonylamino-2 -((((9Z, 12Z) -octadeca-9,12-dienoyloxy) methyl) propane-1,3-diyl dioctadeca-9,12-dienoate (2.08 g, 2.06 mmol, yield 90%) was obtained.
¹ H-NMR (CDCl ₃ ) δ: 0.89 (t, J = 6.7 Hz, 9H), 1.23-1.40 (m, 9H), 1.43 (s, 9H), 1.57-1.66 (m, 14H), 2.05 (q , J = 6.8 Hz, 12H), 2.32 (t, J = 7.6 Hz, 6H), 2.77 (t, J = 6.5 Hz, 6H), 4.34 (s, 6H), 4.81 (br s, 1H), 5.28- 5.43 (m, 12H).
Process 2
(9Z, 9'Z, 12Z, 12'Z) -2- (tert-butoxycarbonylamino-2-(((9Z, 12Z) -octadeca-9,12-dienoyloxy) methyl) propane obtained in Step 1 To a solution of -1,3-diyldioctadeca-9,12-dienoate (2.05 g, 2.03 mmol, 90%) in dichloromethane (10 mL) was added trifluoroacetic acid (2 mL, 26.0 mmol) and stirred at room temperature for 1 hour. Saturated aqueous sodium hydrogen carbonate solution was added to the reaction mixture, and the mixture was extracted with chloroform.The organic layer was dried over anhydrous magnesium sulfate, filtered and concentrated under reduced pressure, and the resulting residue was subjected to amino silica gel column chromatography (hexane / ethyl acetate). = 100/0 to 95/5) and purified by (9Z, 9'Z, 12Z, 12'Z) -2- (hydroxymethyl) -2- (9Z, 12Z) -octadeca-9,12-dienamide Propane-1,3-diyl dioctadeca-9,12-dienoate (1.70 g, 1.84 mmol, 91% yield) was obtained.
ESI-MS m / z: 909 (M + H) ⁺ .
Process 3
(9Z, 9'Z, 12Z, 12'Z) -2- (hydroxymethyl) -2- (9Z, 12Z) -octadeca-9,12-dienamidopropane-1,3-diyl obtained in Step 2 To a solution of dioctadeca-9,12-dienoate (0.8933 g, 0.983 mmol) in dichloromethane (9 mL) (9Z, 12Z) -octadeca-9,12-dienoic acid (Sigma-Aldrich, 2.37 g, 8.45 mmol), 1-Ethyl-3- (3-dimethylaminopropyl) carbodiimide hydrochloride (1.62 g, 8.45 mmol) and N, N-dimethylaminopyridine (0.206 g, 1.69 mmol) were added and stirred at room temperature for 2 hours. Concentrated under reduced pressure. The resulting residue was purified by amino silica gel column chromatography (hexane / ethyl acetate = 90 / 10-75 / 25) to obtain (9Z, 9'Z, 12Z, 12'Z) -2-((4- (dimethyl Amino) butanoyloxy) methyl) -2- (9Z, 12Z) -octadeca-9,12-dienamidopropane-1,3-diyl dioctadeca-9,12-dienoate (0.900 g, 0.881 mmol, 90% yield) )
ESI-MS m / z: 1022 (M + H) ⁺ .
Process 4
(9Z, 9'Z, 12Z, 12'Z) -2-((4- (dimethylamino) butanoyloxy) methyl) -2- (9Z, 12Z) -octadeca-9,12 obtained in Step 3 Methyl iodide (0.493 mL) was added to a chloroform (4 mL) solution of -dienamidepropane-1,3-diyl dioctadeca-9,12-dienoate (0.805 g, 0.788 mmol), and the mixture was stirred at room temperature overnight. The reaction solution was concentrated under reduced pressure. Dissolve the residue in a small amount of methanol-chloroform (1: 1) and load it onto an ion exchange resin (Dowex (TM) 1x-2 100 mesh, Cl type, approx. 20 times volume, pre-washed with water and methanol). -Eluted with chloroform (1: 1). The eluate was concentrated under reduced pressure, and the resulting residue was purified by silica gel column chromatography (chloroform / methanol = 90 / 10-80 / 20) to give the title compound (0.740 g, 0.690 mmol, yield 88%). Obtained.
ESI-MS m / z: 1036 (M) ⁺ ; ¹ H-NMR (CDCl ₃ ) δ: 0.88 (t, J = 6.4 Hz, 9H), 1.21-1.40 (m, 45H), 1.54-1.65 (m, 6H), 2.01-2.08 (m, 12H), 2.09-2.19 (m, 2H), 2.24 (t, J = 7.4 Hz, 2H), 2.32 (t, J = 7.5 Hz, 4H), 2.57 (t, J = 6.2 Hz, 2H), 2.77 (t, J = 6.3 Hz, 6H), 3.41 (s, 9H), 3.84 (t, J = 8.3 Hz, 2H), 4.37-4.50 (m, 6H), 5.28-5.43 (m, 12H), 6.72 (br s, 1H).

4- (1,3-Bis ((9Z, 12Z) -octadeca-9,12-dienoyloxy) -2-(((9Z, 12Z) -octadeca-9,12-dienoyloxy) methyl) propan-2-ylamino) -N, N, N-Trimethyl-4-oxobutane-1-aminium chloride (Compound II-22)
Process 1
Tert-Butyldimethylsilyl chloride (Sigma-Aldrich) in a solution of 2-amino-2- (hydroxymethyl) -1,3-propanediol (Wako Pure Chemical Industries, 7.41 g, 61.2 mmol) in dichloromethane (60 mL) , 9.43 g, 60.7 mmol) and imidazole (Nacalai Tesque, 5.51 g, 80.9 mmol) were added and stirred at room temperature overnight. Saturated saline was added to the reaction solution, and the mixture was extracted with chloroform. The organic layer was dried over anhydrous magnesium sulfate, filtered and concentrated under reduced pressure. The resulting residue was purified by amino silica gel column chromatography (hexane / ethyl acetate = 100/0 to 95/5) to give 6-((tert-butyldimethylsilyloxy) methyl) -2,2,3,3, 9,9,10,10-octamethyl-4,8-dioxa-3,9-disilaundecan-6-amine (3.80 g, 8.19 mmol, 40% yield) was obtained.
ESI-MS m / z: 464 (M + H) ⁺ .
Process 2
6-((tert-butyldimethylsilyloxy) methyl) -2,2,3,3,9,9,10,10-octamethyl-4,8-dioxa-3,9-disila obtained in step 1 To a solution of undecan-6-amine (1.28 g, 2.76 mmol) in dichloromethane (10 mL) was added 4- (dimethylamino) butyric acid hydrochloride (Sigma-Aldrich, 0.708 g, 4.14 mmol), 1-ethyl-3- ( Add 3-dimethylaminopropyl) carbodiimide hydrochloride (0.810 g, 4.14 mmol), N, N-dimethylaminopyridine (0.0170 g, 0.138 mmol) and N, N-diisopropylethylamine (1.45 mL, 8.31 mmol) at room temperature. Stir overnight. Concentrated under reduced pressure. The resulting residue was purified by amino silica gel column chromatography (hexane / ethyl acetate = 95/5 to 90/10) to give N- (6-((tert-butyldimethylsilyloxy) methyl) -2,2,3 , 3,9,9,10,10-Octamethyl-4,8-dioxa-3,9-disilaundecan-6-yl) -4- (dimethylamino) butanamide (1.22 g, 2.11 mmol, 76% yield) )
ESI-MS m / z: 578 (M + H) ⁺ .
Process 3
N- (6-((tert-butyldimethylsilyloxy) methyl) -2,2,3,3,9,9,10,10-octamethyl-4,8-dioxa-3,9 obtained in step 2 -Disilaundecan-6-yl) -4- (dimethylamino) butanamide (1.08 g, 1.87 mmol) in tetrahydrofuran (10 mL) in tetrabutylammonium fluoride (Tokyo Chemical Industry Co., Ltd., approx. 1 mol / L tetrahydrofuran solution) , 7.49 mL, 7.49 mmol) was added, and the mixture was stirred at room temperature for 2 hours. (9Z, 12Z) -octadeca-9,12-dienoic acid (2.05 g, 7.31 mmol), 1-ethyl-3- (3-dimethylaminopropyl) carbodiimide hydrochloride (1.44 g, 7.51 mmol) N, N-dimethylaminopyridine (0.0340 g, 0.278 mmol) was added, and the mixture was stirred overnight at room temperature. Concentrated under reduced pressure. The resulting residue was purified by amino silica gel column chromatography (hexane / ethyl acetate = 60 / 40-50 / 50) to obtain (9Z, 9'Z, 12Z, 12'Z) -2- (4- (dimethylamino ) Butanamide) -2-(((9Z, 12Z) -octadeca-9,12-dienoyloxy) methyl) propane-1,3-diyl dioctadeca-9,12-dienoate (0.405 g, 0.396 mmol, 21% yield) Got.
ESI-MS m / z: 1022 (M + H) ⁺ .
Process 4
(9Z, 9'Z, 12Z, 12'Z) -2- (4- (dimethylamino) butanamide) -2-(((9Z, 12Z) -octadeca-9,12-dienoyloxy) obtained in Step 3 Methyl iodide (Tokyo Chemical Industry Co., Ltd., 0.200 mL) was added to a solution of (methyl) propane-1,3-diyldioctadeca-9,12-dienoate (0.335 g, 0.328 mmol) in chloroform (3 mL), and 2 at room temperature. Stir for hours. The reaction solution was concentrated under reduced pressure. Dissolve the residue in a small amount of methanol-chloroform (1: 1) and load it onto an ion exchange resin (Dowex (TM) 1x-2 100 mesh, Cl type, approx. 20 times volume, pre-washed with water and methanol). -Eluted with chloroform (1: 1). The eluate was concentrated under reduced pressure, and the resulting residue was purified by silica gel column chromatography (chloroform / methanol = 90 / 10-80 / 20) to give the title compound (0.324 g, 0.302 mmol, 92% yield). Obtained.
ESI-MS m / z: 1036 (M) ⁺ ; ¹ H-NMR (CDCl ₃ ) δ: 0.89 (t, J = 7.0 Hz, 9H), 1.23-1.40 (m, 45H), 1.55-1.64 (m, 6H), 2.01-2.12 (m, 14H), 2.34 (t, J = 7.6 Hz, 6H), 2.43 (t, J = 6.3 Hz, 2H), 2.77 (t, J = 6.6 Hz, 6H), 3.37 ( s, 9H), 3.77-3.83 (m, 2H), 4.43 (s, 6H), 5.28-5.42 (m, 12H), 6.62 (br s, 1H).

2- (1,3-bis ((9Z, 12Z) -octadeca-9,12-dienoyloxy) -2-(((9Z, 12Z) -octadeca-9,12-dienoyloxy) methyl) propan-2-ylamino) -N, N, N-Trimethyl-2-oxoethanaminium chloride (Compound II-23)
In the same manner as in Example 29, N, N-dimethylglycine (manufactured by Tokyo Chemical Industry Co., Ltd.) was used instead of 4- (dimethylamino) butyric acid hydrochloride, and the title compound (0.356 g, 0.341 mmol, overall yield) 17%) was obtained.
ESI-MS m / z: 1008 (M) ⁺ ; ¹ H-NMR (CDCl ₃ ) δ: 0.89 (t, J = 7.0 Hz, 9H), 1.23-1.40 (m, 44H), 1.54-1.64 (m, 26H), 2.01-2.08 (m, 12H), 2.35 (t, J = 7.6 Hz, 6H), 2.77 (t, J = 6.8 Hz, 6H), 3.40 (s, 9H), 4.46 (s, 6H), 4.70 (s, 2H), 5.28-5.42 (m, 12H), 9.54 (br s, 1H).

4-((6Z, 9Z, 29Z, 32Z) -20-hydroxy-20-((9Z, 12Z) -octadeca-9,12-dienyl) octatriconta-6,9,29,32-tetraene-19- (Iloxy) -N, N, N-trimethyl-4-oxobutane-1-aminium chloride (Compound III-1)
(6Z, 9Z, 29Z, 32Z) -20-hydroxy-20-((9Z, 12Z) -octadeca-9, obtained by a method according to the method described in U.S. Patent Application Publication No. 2012/0172411 12-Dienyl) octatriconta-6,9,29,32-tetraen-19-yl 4- (dimethylamino) butanoate (0.144 g, 0.156 mmol) was used in the same manner as in Step 2 of Example 1. The title compound (0.146 g, 0.150 mmol, yield 96%) was obtained.
ESI-MS m / z: 935 (M) ⁺ ; ¹ H-NMR (CDCl ₃ ) δ: 0.89 (t, J = 6.9 Hz, 9H), 1.16-1.79 (m, 60H), 1.98-2.17 (m, 15H), 2.52-2.59 (m, 2H), 2.77 (t, J = 6.6 Hz, 6H), 3.44 (s, 9H), 3.69-3.81 (m, 2H), 4.94-4.98 (m, 1H), 5.29 -5.42 (m, 12H).

(6Z, 9Z, 28Z, 31Z) -N, N-dimethyl-N-((9Z, 12Z) -octadeca-9,12-dienyl) heptatriconta-6,9,28,31-tetraene-19-aminium Chloride (Compound IV-1)
Process 1
(6Z, 9Z, 28Z, 31Z) -heptatriconta-6,9,28,31-tetraen-19-one (0.194 g, obtained by a method according to the method described in WO2010 / 042877 Methylamine (manufactured by Tokyo Chemical Industry Co., Ltd., about 40% methanol solution, 0.110 mL, 1.1 mmol) and acetic acid (0.063 mL, 1.1 mmol) were added to a solution of 0.368 mmol) in 1,2-dichloroethane (2 mL). Further, sodium triacetoxyborohydride (0.117 g, 0.552 mmol) was added, followed by stirring at room temperature for 2 hours. Methylamine (about 40% methanol solution, 0.110 mL, 1.1 mmol), acetic acid (0.063 mL, 1.1 mmol), and sodium triacetoxyborohydride (0.117 g, 0.552 mmol) were added to the reaction solution, and the mixture was stirred for 2 hours. A saturated aqueous sodium hydrogen carbonate solution was added to the reaction mixture, and the mixture was extracted twice with hexane. The organic layers were combined, dried over anhydrous magnesium sulfate, filtered, and concentrated under reduced pressure. The obtained residue was purified by column chromatography (chloroform / methanol = 100/0 to 90/10) and (6Z, 9Z, 28Z, 31Z) -N-methylheptatria contour-6,9,28,31- Tetraene-19-amine (0.121 g, 0.223 mmol, 61% yield) was obtained.
ESI-MS m / z: 543 (M + H) ⁺ .
Process 2
(9Z, 12Z)-(6Z, 9Z, 28Z, 31Z) -N-methylheptatriconta-6,9,28,31-tetraen-19-amine (0.121 g, 0.223 mmol) obtained in Step 1 Octadeca-9,12-dienyl methanesulfonate (0.154 g, 0.446 mmol) and 50% aqueous sodium hydroxide solution (0.107 g, 1.34 mmol) were added, and the mixture was stirred at 135 ° C. for 2 hours in an oil bath. The reaction solution was cooled to room temperature, saturated brine was added, and the mixture was washed with hexane. The organic layer was dried over anhydrous magnesium sulfate, filtered and concentrated under reduced pressure. The obtained residue was purified by silica gel column chromatography (hexane / ethyl acetate = 100/0 to 85/15) to give (6Z, 9Z, 28Z, 31Z) -N-methyl-N-((9Z, 12Z ) -Octadeca-9,12-dienyl) heptatriconta-6,9,28,31-tetraen-19-amine (0.139 g, 0.175 mmol, yield 79%) was obtained.
ESI-MS m / z: 792 (M + H) ⁺ .
Process 3
(6Z, 9Z, 28Z, 31Z) -N-methyl-N-((9Z, 12Z) -octadeca-9,12-dienyl) heptatriconta-6,9,28,31-tetraene obtained in Step 2 The title compound (0.114 g, 0.135 mmol, yield 77%) was obtained in the same manner as in Example 1, Step 2 using -19-amine (0.139 g, 0.175 mmol).
ESI-MS m / z: 806 (M) ⁺ ; ¹ H-NMR (CDCl ₃ ) δ: 0.89 (t, J = 7.0 Hz, 9H), 1.26-1.56 (m, 54H), 1.65-1.73 (m, 2H), 1.80-1.88 (m, 2H), 2.05 (q, J = 7.0 Hz, 12H), 2.77 (t, J = 6.3 Hz, 6H), 3.22-3.27 (m, 1H), 3.31 (s, 6H ), 3.58-3.62 (m, 2H), 5.29-5.42 (m, 12H).

N, N, N-trimethyl-3- (palmitoyloxy) -2,2-bis ((palmitoyloxy) methyl) propane-1-aminium chloride (Compound II-24)
Process 1
2-((Dimethylamino) methyl) -2- (hydroxymethyl) propane-1,3-diol (0.420 g, 2.57 mmol) of 1,2-dichloroethane (5 mL) obtained in Step 1 of Example 24 After adding pyridine (3.12 mL, 38.6 mmol) to the solution, palmitoyl chloride (manufactured by Tokyo Chemical Industry Co., Ltd., 6.22 mL, 20.6 mmol) was added at room temperature, and the mixture was stirred at 70 ° C. for 2 hours. The reaction mixture was cooled to room temperature, water was added, and the mixture was extracted with ethyl acetate. The organic layer was washed with saturated brine, dried over anhydrous magnesium sulfate, and filtered. After concentration under reduced pressure, the resulting residue was purified by silica gel column chromatography (chloroform) to give 2-((dimethylamino) methyl) -2-((palmitoyloxy) methyl) propane-1,3-diyl dipalmitate. (0.650 g, 0.740 mmol, 29% yield) was obtained.
ESI-MS m / z: 879 (M + H) ⁺
Process 2
Using 2-((dimethylamino) methyl) -2-((palmitoyloxy) methyl) propane-1,3-diyl dipalmitate (0.65 g, 0.74 mmol) obtained in Step 1, as in Step 1 of Example 1. To give the title compound (0.056 g, 0.060 mmol, 8% yield).
ESI-MS m / z: 893 (M) ⁺ ; ¹ H-NMR (CDCl ₃ ) δ: 0.88 (t, J = 7.0 Hz, 9H), 1.21-1.34 (m, 72H), 1.54-1.64 (m, 6H), 2.38 (t, J = 7.6 Hz, 6H), 3.62 (s, 9H), 3.95 (s, 2H), 4.29 (s, 6H).

N, N, N-trimethyl-3- (tetradecanoyloxy) -2,2-bis ((tetradecanoyloxy) methyl) propane-1-aminium chloride (compound II-25)
In the same manner as in Example 33, myristoyl chloride (Wako Pure Chemical Industries) was used in place of palmitoyl chloride to obtain the title compound (0.045 g, 0.053 mmol, overall yield 4%).
ESI-MS m / z: 809 (M) ⁺ ; ¹ H-NMR (CDCl ₃ ) δ: 0.88 (t, J = 6.8 Hz, 9H), 1.21-1.34 (m, 60H), 1.54-1.64 (m, 6H), 2.38 (t, J = 7.6 Hz, 6H), 3.64 (s, 9H), 3.96 (s, 2H), 4.29 (s, 6H).

3- (Dodecanoyloxy) -2,2-bis ((dodecanoyloxy) methyl) -N, N, N-trimethylpropane-1-aminium chloride (Compound II-26)
In the same manner as in Example 33, lauroyl chloride (manufactured by Tokyo Chemical Industry Co., Ltd.) was used instead of palmitoyl chloride to obtain the title compound (0.085 g, 0.112 mmol, overall yield 9%).
ESI-MS m / z: 725 (M) ⁺ ; ¹ H-NMR (CDCl ₃ ) δ: 0.88 (t, J = 6.8 Hz, 9H), 1.19-1.34 (m, 48H), 1.54-1.64 (m, 6H), 2.38 (t, J = 7.6 Hz, 6H), 3.66 (s, 9H), 3.97 (s, 2H), 4.30 (s, 6H).

(Z) -N, N, N-Trimethyl-3,3-bis ((oleoyloxy) methyl) henicosa-12-en-1-aminium chloride (Compound II-27)
Process 1
Dimethyl malonate (1.00 g, 7.57 mmol) was dissolved in acetonitrile (25 mL) and (Z) -oct-9-en-1-yl methanesulfonate (3.15 g, 9.08 mmol), cesium carbonate (4.93 g, 15.1 mmol) and tetrabutylammonium iodide (3.35 g, 9.08 mmol) were added, and the mixture was stirred at 60 ° C. for 1 hour. Water was added to the reaction mixture, and the mixture was extracted with hexane. The organic layer was washed with saturated brine, dried over anhydrous magnesium sulfate, filtered, and concentrated under reduced pressure. The resulting residue was purified by silica gel column chromatography (hexane / ethyl acetate = 90/10 to 70/30) to give (Z) -dimethyl 2- (octadeca-9-en-1-yl) malonate (2.54 g, 6.64 mmol, yield 88%).
ESI-MS m / z: 383 (M + H) ⁺
Process 2
(Z) -dimethyl 2- (octadeca-9-en-1-yl) malonate (0.500 g, 1.31 mmol) obtained in step 1 was dissolved in toluene (6 mL), and sodium hydride (oil-based oil was added under ice cooling. , 60%, 0.209 g, 5.23 mmol) and stirred until no foaming occurred. Subsequently, 2-chloro-N, N-dimethylethanamine hydrochloride (manufactured by Tokyo Chemical Industry Co., Ltd., 0.377 g, 2.61 mmol) was added, and the mixture was stirred at 100 ° C. for 2 hours. Water was added to the reaction mixture under ice cooling, and the mixture was extracted with chloroform. The organic layer was washed with saturated brine, dried over anhydrous magnesium sulfate, filtered, and concentrated under reduced pressure. The obtained residue was purified by silica gel column chromatography (chloroform / methanol = 99/1 to 90/10) to give (Z) -dimethyl 2- (2- (dimethylamino) ethyl) -2- (octadeca- 9-en-1-yl) malonate (0.258 g, 0.569 mmol, 44% yield) was obtained.
ESI-MS m / z: 454 (M + H) ⁺
Process 3
(Z) -Dimethyl 2- (2- (dimethylamino) ethyl) -2- (octadeca-9-en-1-yl) malonate obtained in Step 2 in the same manner as in Step 2 of Example 19. 0.250 g, 0.551 mmol) and (Z) -2- (2- (dimethylamino) ethyl) -2- (octadeca-9-en-1-yl) propane-1,3-diol (0.220 g, 0.553 mmol, quantitative).
ESI-MS m / z: 398 (M + H) ⁺
Process 4
(Z) -2- (2- (dimethylamino) ethyl) -2- (octadeca-9-en-1-yl) propane-1,3-diol (0.220 g, 0.553 mmol) obtained in Step 3 was Dissolve in dichloromethane (2 mL), add N, N-diisopropylethylamine (0.386 mL, 2.21 mmol), add oleoyl chloride (Sigma-aldrich, 0.457 mL, 1.38 mmol) under ice-cooling, and add 10 at room temperature. Stir for minutes. Water was added to the reaction mixture, and the mixture was extracted with hexane. The organic layer was washed with saturated brine, dried over anhydrous magnesium sulfate, filtered, and concentrated under reduced pressure. The obtained residue was purified by silica gel column chromatography (hexane / ethyl acetate = 90 / 10-50 / 50) to give (Z) -2- (2- (dimethylamino) ethyl) -2-((Z ) -Octadeca-9-en-1-yl) propane-1,3-diyl dioleate (0.280 g, 0.302 mmol, yield 55%) was obtained.
ESI-MS m / z: 927 (M + H) ⁺
Process 5
(Z) -2- (2- (Dimethylamino) ethyl) -2-((Z) -octadeca-9-en-1-yl) propane-1,3-diyl dioleate obtained in Step 4 (0.280 g , 0.302 mmol), and the title compound (0.199 g, 0.204 mmol, yield 67%) was obtained in the same manner as in Step 1 of Example 1.
ESI-MS m / z: 941 (M) ⁺ ; ¹ H-NMR (CDCl ₃ ) δ: 0.88 (t, J = 7.0 Hz, 9H), 1.22-1.39 (m, 66H), 1.55-1.65 (m, 4H), 1.70-1.78 (m, 2H), 1.98-2.06 (m, 12H), 2.33 (t, J = 7.6 Hz, 4H), 3.46 (s, 9H), 3.58-3.65 (m, 2H), 3.93 -4.03 (m, 4H), 5.29-5.39 (m, 6H).

(Z) -N, N, N-Trimethyl-4,4-bis ((oleoyloxy) methyl) docosa-13-en-1-aminium chloride (Compound II-28)
Process 1
In the same manner as in Step 2 of Example 36, instead of 2-chloro-N, N-dimethylethanamine hydrochloride, 3-chloro-N, N-dimethylpropan-1-amine hydrochloride (manufactured by Tokyo Chemical Industry Co., Ltd.) ) To give (Z) -dimethyl 2- (3- (dimethylamino) propyl) -2- (octadeca-9-en-1-yl) malonate (0.210 g, 0.449 mmol, 34% yield) .
ESI-MS m / z: 468 (M + H) ⁺
Process 2
In the same manner as in Steps 3, 4, and 5 of Example 36, instead of (Z) -dimethyl 2- (2- (dimethylamino) ethyl) -2- (octadeca-9-en-1-yl) malonate Using (Z) -dimethyl 2- (3- (dimethylamino) propyl) -2- (octadeca-9-en-1-yl) malonate (0.210 g, 0.449 mmol) obtained in Step 1, the title compound ( 0.042 g, 0.042 mmol, 9% yield).
ESI-MS m / z: 955 (M) ⁺ ; ¹ H-NMR (CDCl ₃ ) δ: 0.88 (t, J = 6.8 Hz, 9H), 1.22-1.36 (m, 68H), 1.56-1.64 (m, 4H), 1.72-1.82 (m, 2H), 1.96-2.07 (m, 12H), 2.32 (t, J = 7.5 Hz, 4H), 3.38 (s, 9H), 3.39-3.46 (m, 2H), 3.93 (d, J = 11.2 Hz, 2H), 3.99 (d, J = 11.2 Hz, 2H), 5.28-5.40 (m, 6H).

N, N, N-trimethyl-3- (stearoyloxy) -2,2-bis ((stearoyloxy) methyl) propane-1-aminium chloride (compound II-29)
In the same manner as in Example 33, stearoyl chloride (manufactured by Tokyo Chemical Industry Co., Ltd.) was used instead of palmitoyl chloride to obtain the title compound (0.085 g, 0.112 mmol, overall yield 6%).
ESI-MS m / z: 977 (M) ⁺ ; ¹ H-NMR (CDCl ₃ ) δ: 0.88 (t, J = 6.8 Hz, 9H), 1.21-1.37 (m, 84H), 1.54-1.64 (m, 6H), 2.38 (t, J = 7.6 Hz, 6H), 3.65 (s, 9H), 3.96 (s, 2H), 4.30 (s, 6H).

N, N, N-trimethyl-3-oleamide-2,2-bis ((oleoyloxy) methyl) propane-1-aminium chloride (Compound II-30)
Process 1
2-((Dimethylamino) methyl) -2- (hydroxymethyl) propane-1,3-diol (0.410 g, 2.51 mmol) obtained in Step 1 of Example 24 was added to dichloromethane (5 mL), pyridine (5.08). (mL, 62.8 mmol). Under ice-cooling, oleoyl chloride (1.25 mL, 3.77 mmol) was added. Water was added to the reaction mixture, and the mixture was extracted with chloroform. The organic layer was washed with saturated brine, dried over anhydrous magnesium sulfate, and filtered. The residue obtained by concentration under reduced pressure was purified by silica gel column chromatography (chloroform / methanol = 99/1 to 90/10) to give (Z) -2-((dimethylamino) methyl) -2- ( Hydroxymethyl) propane-1,3-diyl dioleate (0.190 g, 0.275 mmol, yield 11%) was obtained.
ESI-MS m / z: 693 (M + H) ⁺
Process 2
(Z) -2-((Dimethylamino) methyl) -2- (hydroxymethyl) propane-1,3-diyldiolate (0.190 g, 2.51 mmol) obtained in step 1 was dissolved in toluene (2 mL). Then, diphenylphosphoric acid azide (manufactured by Tokyo Chemical Industry Co., Ltd., 0.118 mL, 0.549 mmol) and 1,8-diazabicyclo [5.4.0] -7-undecene (0.0830 mL, 0.549 mmol) were added at room temperature, and the mixture was stirred for 1 hour. Since the progress of the reaction was insufficient, diphenylphosphoric acid azide (0.118 mL, 0.549 mmol) was added, and the mixture was stirred with heating at 80 ° C. for 3 hr. The reaction mixture was cooled to room temperature, water was added, and the mixture was extracted with ethyl acetate. The organic layer was washed with saturated brine, dried over anhydrous magnesium sulfate, and filtered. Concentration under reduced pressure, and the resulting residue was purified by silica gel column chromatography (hexane / ethyl acetate = 95 / 5-60 / 40) to give (Z) -2- (azidomethyl) -2-((dimethylamino ) Methyl) propane-1,3-diyl dioleate (0.135 g, 0.188 mmol, 69% yield) was obtained.
ESI-MS m / z: 718 (M + H) ⁺
Process 3
(Z) -2- (azidomethyl) -2-((dimethylamino) methyl) propane-1,3-diyldiolate (0.135 g, 2.51 mmol) obtained in step 2 was added to tetrahydrofuran (1 mL) and water (0.1 In a mixed solution, triphenylphosphine (manufactured by Junsei Chemical Co., Ltd., 0.0740 g, 0.282 mmol) was added and stirred for 3 hours. Water was added to the reaction mixture, and the mixture was extracted with ethyl acetate. The organic layer was washed with saturated brine, dried over anhydrous magnesium sulfate, and filtered. Concentration under reduced pressure gave a crude product of (Z) -2- (aminoethyl) -2-((dimethylamino) methyl) propane-1,3-diyldiolate (0.130 g, 0.188 mmol, 100% yield) )
ESI-MS m / z: 691 (M + H) ⁺
Process 4
To a solution of (Z) -2- (aminoethyl) -2-((dimethylamino) methyl) propane-1,3-diyl dioleate (0.130 g, 0.188 mmol) obtained in step 3 in dichloromethane (2 mL), N, N-diisopropylethylamine (0.0990 mL, 0.564 mmol) was added, oleoyl chloride (0.0850 g, 0.282 mmol) was added under ice cooling, and the mixture was stirred at room temperature for 1 hr. Water was added to the reaction mixture, and the mixture was extracted with hexane. The organic layer was washed with saturated brine, dried over anhydrous magnesium sulfate, and filtered. Concentration under reduced pressure, and the resulting residue was purified by silica gel column chromatography (hexane / ethyl acetate = 97 / 3-60 / 40) to give (Z) -2-((dimethylamino) methyl) -2- (Oleamidomethyl) propane-1,3-diyl dioleate (0.105 g, 0.110 mmol, yield 58%) was obtained.
ESI-MS m / z: 956 (M + H) ⁺
Process 5
Using (Z) -2-((dimethylamino) methyl) -2- (oleamidomethyl) propane-1,3-diyldiolate (0.105 g, 0.110 mmol) obtained in step 4, Example 1 step 2 In the same manner as described above, the title compound (0.0480 g, 0.0480 mmol, yield 43%) was obtained.
ESI-MS m / z: 970 (M) ⁺ ; ¹ H-NMR (CDCl ₃ ) δ: 0.88 (t, J = 6.8 Hz, 9H), 1.21-1.37 (m, 60H), 1.56-1.65 (m, 6H), 1.96-2.05 (m, 12H), 2.36 (t, J = 7.6 Hz, 6H), 3.51 (s, 9H), 3.51-3.56 (m, 2H), 4.02 (s, 2H), 4.20 (d , J = 12.2 Hz, 2H), 4.30 (d, J = 12.2 Hz, 2H), 5.27-5.40 (m, 6H), 8.11-8.20 (m, 1H).

N, N, N-trimethyl-4- (oleoyloxy) -3,3-bis (oleoyloxymethyl) butane-1-aminium chloride (Compound II-31)
Process 1
To a solution of 2- (bromomethyl) -2- (hydroxymethyl) propane-1,3-diol (Tokyo Chemical Industry Co., Ltd., 1.00 g, 5.02 mmol) in tetrahydrofuran (10 mL) was added tert-butyldimethylchlorosilane (Tokyo Chemical Industry Co., Ltd.). (3,79 g, 25.1 mmol), imidazole (manufactured by Nacalai Tesque, 3.42 g, 50.2 mmol) and N, N-dimethylaminopyridine (0.061 g, 0.502 mmol) were added, and the mixture was stirred at room temperature overnight. A saturated aqueous sodium hydrogen carbonate solution was added to the reaction mixture, and the mixture was extracted twice with hexane. The organic layer was dried over anhydrous magnesium sulfate and concentrated under reduced pressure. The residue was purified by silica gel column chromatography (hexane) to give 6- (bromomethyl) -6-((tert-butyldimethylsilyloxy) methyl) -2,2,3,3,9,9,10,10- Octamethyl-4,8-dioxa-3,9-disilaundecane (2.50 g, 4.61 mmol, 92%) was obtained.
¹ H-NMR (CDCl3) δ: 0.04 (s, 18H), 0.89 (s, 27H), 3.41 (s, 2H), 3.49 (s, 6H).
Process 2
6- (Bromomethyl) -6-((tert-butyldimethylsilyloxy) methyl) -2,2,3,3,9,9,10,10-octamethyl-4,8-dioxa- obtained in Step 1 To a solution of 3,9-disilaundecane (1.849 g, 3.41 mmol) in dimethyl sulfoxide (10 mL) was added sodium cyanide (Nacalai Tesque, 0.529 g, 10.8 mmol) and stirred at 85 ° C for 3 days. did. After cooling to room temperature, the reaction mixture was diluted with hexane, washed with water and then with saturated brine, dried over anhydrous magnesium sulfate, filtered, and concentrated under reduced pressure. The obtained residue was purified by silica gel column chromatography (hexane / ethyl acetate = 90/10) to give 4- (tert-butyldimethylsilyloxy) -3,3-bis ((tert-butyldimethylsilyloxy) Methyl) butanenitrile (1.35 g, 2.77 mmol, 81% yield) was obtained.
ESI-MS m / z: 489 (M + H) ⁺ ; ¹ H-NMR (CDCl ₃ ) δ: 0.05 (s, 18H), 0.89 (s, 27H), 2.34 (s, 2H), 3.51 (s, 6H).
Process 3
To a solution of 4- (tert-butyldimethylsilyloxy) -3,3-bis ((tert-butyldimethylsilyloxy) methyl) butanenitrile (1.34 g, 2.75 mmol) obtained in step 2 in tetrahydrofuran (10 mL) Then, lithium aluminum hydride (0.104 g, 2.75 mmol) was added under ice cooling, and the mixture was stirred at room temperature for 2 hr. Water (0.495 mL, 27.5 mmol) and sodium fluoride (3.46 g, 82.0 mmol) were added to the reaction solution, and the mixture was stirred at room temperature overnight. Insoluble materials were removed by Celite filtration, and the filtrate was concentrated. The obtained residue was purified by amino silica gel column chromatography (ethyl acetate) to give 4- (tert-butyldimethylsilyloxy) -3,3-bis ((tert-butyldimethylsilyloxy) methyl) butane-1 -Amine (0.435 g, 0.884 mmol, Yield 32%) was obtained.
ESI-MS m / z: 493 (M + H) ⁺ ; ¹ H-NMR (CDCl ₃ ) δ: 0.02 (s, 18H), 0.88 (s, 27H), 1.38-1.43 (m, 2H), 2.71- 2.75 (m, 2H), 3.40 (s, 6H).
Process 4
4- (tert-Butyldimethylsilyloxy) -3,3-bis ((tert-butyldimethylsilyloxy) methyl) butan-1-amine (0.200 g, 0.407 mmol) of 1,2- To a dichloroethane (3 mL) solution were added 38% aqueous formaldehyde solution (0.295 mL) and sodium triacetoxyborohydride (0.431 g, 2.03 mmol), and the mixture was stirred at room temperature overnight. A saturated aqueous sodium hydrogen carbonate solution was added to the reaction mixture, and the mixture was extracted twice with chloroform. The organic layer was dried over anhydrous magnesium sulfate and concentrated under reduced pressure. The residue was purified by silica gel column chromatography (chloroform / methanol = 90/10) to give 4- (tert-butyldimethylsilyloxy) -3,3-bis ((tert-butyldimethylsilyloxy) methyl) -N Thus, a crude product of N-dimethylbutan-1-amine was obtained.
Tetrahydrofuran (2 mL) and tetrabutylammonium fluoride (manufactured by Tokyo Chemical Industry Co., Ltd., about 1 mol / L tetrahydrofuran solution, 2.06 mL, 2.06 mmol) were added to the resulting crude product, and the mixture was stirred at room temperature for 5 hours, then 60 Stir overnight at ° C. Saturated aqueous sodium hydrogen carbonate solution was added to the reaction mixture, and the mixture was washed twice with chloroform. The aqueous layer was concentrated under reduced pressure. Acetone (2 mL), sodium hydroxide (manufactured by Wako Pure Chemical Industries, 2 mol / L aqueous solution, 3 mL, 6 mmol) and oleoyl chloride (0.681 mL, 2.06 mmol) are added to the resulting residue, and the mixture is brought to room temperature. And stirred for 3 hours. To the reaction solution was added oleoyl chloride (0.681 mL, 2.06 mmol), and the mixture was stirred at 60 ° C. overnight. After cooling to room temperature, water was added to the reaction mixture, and the mixture was extracted twice with chloroform. The organic layer was dried over anhydrous magnesium sulfate, filtered and concentrated under reduced pressure. The obtained residue was purified by silica gel column chromatography (chloroform / methanol = 90/10) to obtain (Z) -2- (2- (dimethylamino) ethyl) -2- (oleyloxymethyl) propane-1 Thus, a crude product of 3-diyldiolate was obtained. The resulting crude product was dissolved in a small amount of methanol-chloroform (9: 1) and loaded onto an ion exchange resin (Waters, PoraPack Rxn CX, pre-washed with methanol) and ammonia (Sigma-Aldirch). And 2 mol / L methanol solution). The eluate was concentrated under reduced pressure, and (Z) -2- (2- (dimethylamino) ethyl) -2- (oleyloxymethyl) propane-1,3-diyldiolate (0.387 g, 0.399 mmol, yield 98). %).
ESI-MS m / z: 971 (M + H) ⁺ ; ¹ H-NMR (CDCl ₃ ) δ: 0.88 (t, J = 7.0 Hz, 9H), 1.21-1.38 (m, 62H), 1.54-1.65 ( m, 6H), 1.97-2.04 (m, 12H), 2.20 (s, 6H), 2.25-2.32 (m, 8H), 4.04 (s, 6H), 5.29-5.39 (m, 6H).
Process 5
In the same manner as in Example 1, Step 2, 2, Z'-2- (2) was obtained in Step 4 instead of 2,2 ′, 2 ''-nitrilotris (ethane-2,1-diyl) trioleate. -(Dimethylamino) ethyl) -2- (oleyloxymethyl) propane-1,3-diyldiolate (0.109 g, 0.112 mol) and the title compound (0.0642 g, 0.0630 mol, yield 56%) Obtained.
ESI-MS m / z: 986 (M) ⁺ ; ¹ H-NMR (CDCl ₃ ) δ: 0.88 (t, J = 6.8 Hz, 9H), 1.23-1.37 (m, 74H), 1.55-1.64 (m, 65H), 1.78-1.84 (m, 2H), 1.95-2.06 (m, 13H), 2.35 (t, J = 7.6 Hz, 6H), 3.42 (s, 8H), 3.70-3.77 (m, 2H), 4.08 (s, 6H), 5.29-5.39 (m, 6H).

N, N, N-trimethyl-2- (3- (oleoyloxy) -2,2-bis ((oleoyloxy) methyl) propoxy) -2-oxyethane-1-aminium chloride (Compound II-32)
Process 1
To a solution of 2,2- (dimethyl-1,3-dioxane-5,5-diyl) dimethanol (0.200 g, 1.14 mmol) synthesized in the method described in US Patent No. 8816099 in tetrahydrofuran (5 mL) Triethylamine (0.475 mL, 3.40 mmol) was added, oleoyl chloride (0.854 g, 2.84 mmol) was added under ice cooling, and the mixture was stirred as it was under ice cooling for 1 hour. Water was added to the reaction mixture, and the mixture was extracted with hexane. The organic layer was washed with saturated brine, dried over anhydrous magnesium sulfate, and filtered. Concentration under reduced pressure, and the resulting residue was purified by silica gel column chromatography (hexane / ethyl acetate = 95/5) to give (2,2-dimethyl-1,3-dioxane-5,5-diyl) bis (Methylene) dioleate (0.500 g, 0.709 mmol, 63% yield) was obtained.
ESI-MS m / z: 705 (M + H) ⁺
Process 2
To a solution of (2,2-dimethyl-1,3-dioxane-5,5-diyl) bis (methylene) dioleate (0.500 g, 0.709 mmol) obtained in step 1 in dichloromethane (5 mL), Fluoroacetic acid (2.00 mL, 26.0 mmol) was added in two portions, followed by stirring for 1 hour under ice cooling. A saturated aqueous sodium hydrogen carbonate solution was added to the reaction mixture, and the mixture was extracted with ethyl acetate. The organic layer was washed with saturated brine, dried over anhydrous magnesium sulfate, and filtered. The obtained residue was purified by silica gel column chromatography (hexane / ethyl acetate = 90 / 10-50 / 50) to give 2,2-bis (hydroxymethyl) propane-1,3-diyl dioleate (0.207 g, 0.311 mmol, yield 44%).
ESI-MS m / z: 665 (M + H) ⁺
Process 3
Thionyl chloride (1 mL, 13.7 mmol) was added to N, N-dimethylglycine (manufactured by Tokyo Chemical Industry Co., Ltd., 0.049 g, 0.474 mmol), and the mixture was heated and stirred at 70 ° C. for 30 minutes. The reaction solution was cooled to room temperature and then concentrated under reduced pressure to obtain a crude product of N, N-dimethylglycinoyl chloride. To a solution of 2,2-bis (hydroxymethyl) propane-1,3-diyldioleate (0.207 g, 0.311 mmol) obtained in step 2 in dichloromethane (5 mL), N, N-diisopropylethylamine (0.110 mL, 0.632 mmol) and the above N, N-dimethylglycinoyl chloride crude product were added and stirred at room temperature for 1 hour. Water was added to the reaction mixture, and the mixture was extracted with ethyl acetate. The organic layer was washed with saturated brine, dried over anhydrous magnesium sulfate, and filtered. The obtained residue was purified by silica gel column chromatography (hexane / ethyl acetate = 90 / 10-40 / 60) to give 2-(((dimethylglycyl) oxy) methyl) -2- (hydroxymethyl) propane. -1,3-Diyl dioleate (0.077 g, 0.103 mmol, 33% yield) was obtained.
ESI-MS m / z: 751 (M + H) ⁺
Process 4
To a solution of 2-(((dimethylglycyl) oxy) methyl) -2- (hydroxymethyl) propane-1,3-diyldiolate (0.0770 g, 0.103 mmol) obtained in step 3 in dichloromethane (3 mL), pyridine was added. (0.0330 mL, 0.411 mmol) was added, oleoyl chloride (0.0620 g, 0.205 mmol) was added under ice cooling, and the mixture was stirred at room temperature for 30 min. Water was added to the reaction mixture, and the mixture was extracted with a mixed solvent of hexane / ethyl acetate = 1/1. The organic layer was washed with saturated brine, dried over anhydrous magnesium sulfate, and filtered. The obtained residue was purified by silica gel column chromatography (hexane / ethyl acetate = 95/5) to give 2-(((dimethylglycyl) oxy) methyl) -2-((oleoyl) methyl) propane- 1,3-diyl dioleate (0.122 g, 0.0600 mmol, yield 59%) was obtained.
ESI-MS m / z: 1015 (M + H) ⁺
Process 5
Using 2-((((dimethylglycyl) oxy) methyl) -2-((oleoyl) methyl) propane-1,3-diyldiolate (0.122 g, 0.060 mmol) obtained in step 4, Example 1 In the same manner as in Step 2, the title compound (0.017 g, 0.016 mmol, yield 27%) was obtained.
ESI-MS m / z: 1029 (M) ⁺ ; ¹ H-NMR (CDCl ₃ ) δ: 0.88 (t, J = 6.8 Hz, 9H), 1.19-1.38 (m, 58H), 1.54-1.66 (m, 6H), 1.98-2.04 (m, 12H), 2.28-2.35 (m, 6H), 3.60 (s, 9H), 4.11 (d, J = 1.8 Hz, 6H), 4.20 (s, 2H), 5.08 (s , 2H), 5.29-5.41 (m, 6H).

N, N, N-trimethyl-1,3-bis (tetradecanoyloxy) -2-((tetradecanoyloxy) methyl) propane-2-aminium chloride (compound II-33)
Process 1
2- (Dimethylamino) -2- (hydroxymethyl) propane-1,3-diol (1.50 g, 10.1 mmol) was added to tetrahydrofuran (10 mL) with pyridine (4.07 mL, 50.3 mmol) and then tetradecanoyl chloride (4.09 mL, 15.1 mmol) was added, and the mixture was heated and stirred at 60 ° C. for 2 hours. Water was added to the reaction mixture, and the mixture was extracted with ethyl acetate. The organic layer was washed with saturated brine, dried over anhydrous magnesium sulfate, and filtered. Concentrated under reduced pressure, and the resulting residue was purified by silica gel column chromatography (chloroform / methanol = 99/1 to 90/10) to give 2- (dimethylamino) -2-((tetradecanoyloxy) methyl. ) Propane-1,3-diyl ditetradecanoate (1.50 g, 1.92 mmol, 19% yield), 2- (dimethylamino) -2- (hydroxymethyl) propane-1,3-diyl ditetradecano Art (0.750 g, 1.32 mmol, 13% yield), and 2- (dimethylamino) -3-hydroxy-2- (midyloxymethyl) propyl tetradecanoate (0.220 g, 0.612 mmol, 6 yield) %) Respectively.
ESI-MS m / z: 781 (M + H) ⁺
ESI-MS m / z: 570 (M + H) ⁺
ESI-MS m / z: 360 (M + H) ⁺
Process 2
Using 2- (dimethylamino) -2-((tetradecanoyloxy) methyl) propane-1,3-diyl ditetradecanoate (1.50 g, 1.92 mmol) obtained in Step 1, Example 1 Step In the same manner as in 2, the title compound (0.530 g, 0.638 mmol, yield 33%) was obtained.
ESI-MS m / z: 795 (M) ⁺ ; ¹ H-NMR (CDCl ₃ ) δ: 0.88 (t, J = 6.8 Hz, 9H), 1.21-1.33 (m, 60H), 1.55-1.65 (m, 6H), 2.37 (t, J = 7.6 Hz, 6H), 3.71 (s, 9H), 4.59 (s, 6H).

N, N, N-trimethyl-1,3-bis (oleoyloxy) -2-((tetradecanoyloxy) methyl) propane-2-aminium chloride (Compound II-34)
Process 1
2- (Dimethylamino) -3-hydroxy-2- (midoxymethyl) propyl tetradecanoate (0.220 g, 0.612 mmol) of 1,2-dichloroethane (3 mL) obtained in Step 1 of Example 42 Pyridine (0.297 mL, 3.67 mmol) was added to the solution, followed by oleoyl chloride (0.552 g, 1.84 mmol), and the mixture was stirred with heating at 60 ° C. for 1 hr. Water was added to the reaction mixture, and the mixture was extracted with ethyl acetate. The organic layer was washed with saturated brine, dried over anhydrous magnesium sulfate, and filtered. The obtained residue was purified by silica gel column chromatography (hexane / ethyl acetate = 98/2 to 85/15) to give 2- (dimethylamino) -2-((tetradecanoyloxy) methyl) propane-1 , 3-Diyl dioleate (0.250 g, 0.281 mmol, 46% yield) was obtained.
ESI-MS m / z: 889 (M + H) ⁺
Process 2
Using 2- (dimethylamino) -2-((tetradecanoyloxy) methyl) propane-1,3-diyldiolate (0.250 g, 0.281 mmol) obtained in Step 1, as in Step 1 of Example 1. The title compound (0.065 g, 0.069 mmol, yield 25%) was obtained.
ESI-MS m / z: 903 (M) ⁺ ; ¹ H-NMR (CDCl ₃ ) δ: 0.88 (t, J = 7.0 Hz, 9H), 1.22-1.38 (m, 60H), 1.56-1.66 (m, 6H), 1.97-2.05 (m, 8H), 2.39 (t, J = 7.6 Hz, 6H), 3.72 (s, 9H), 4.58 (s, 6H), 5.28-5.40 (m, 4H).

N, N, N-trimethyl-1- (oleyloxy) -3- (tetradecanoyloxy) -2-((tetradecanoyloxy) methyl) propane-2-aminium chloride (Compound II-35)
Process 1
2- (Dimethylamino) -2- (hydroxymethyl) propane-1,3-diyl ditetradecanoate (0.750 g, 1.32 mmol) of 1,2-dichloroethane obtained in Step 1 of Example 42 Pyridine (0.532 mL, 6.58 mmol) and then oleoyl chloride (0.792 g, 1.84 mmol) were added to the (3 mL) solution, and the mixture was stirred with heating at 60 ° C. for 1 hr. Water was added to the reaction mixture, and the mixture was extracted with ethyl acetate. The organic layer was washed with saturated brine, dried over anhydrous magnesium sulfate, and filtered. The obtained residue was purified by silica gel column chromatography (hexane / ethyl acetate = 95/5) to give 2- (dimethylamino) -2-((oleyloxy) methyl) propane-1,3-diyl ditetra Decanoate (0.750 g, 0.899 mmol, 68% yield) was obtained.
ESI-MS m / z: 835 (M + H) ⁺
Process 2
Using 2- (dimethylamino) -2-((oleyloxy) methyl) propane-1,3-diyl ditetradecanoate (0.750 g, 0.899 mmol) obtained in Step 1, Example 1 Step 2 and Similarly, the title compound (0.092 g, 0.104 mmol, 12% yield) was obtained.
ESI-MS m / z: 849 (M) ⁺ ; ¹ H-NMR (CDCl3) δ: 0.88 (t, J = 6.8 Hz, 9H), 1.19-1.38 (m, 60H), 1.56-1.65 (m, 6H ), 1.98-2.06 (m, 4H), 2.39 (t, J = 7.6 Hz, 6H), 3.72 (s, 9H), 4.59 (s, 6H), 5.30-5.39 (m, 2H).

N, N, N-trimethyl-3- (tetradecanoyloxy) -2-((tetradecanoyloxy) methyl) -2-(((tetradecylcarbamoyl) oxy) methyl) propane-1-aminium chloride (compound II-36)
Process 1
To a solution of 2-((dimethylamino) methyl) -2- (hydroxymethyl) propane-1,3-diol (0.820 g, 5.02 mmol) obtained in Step 1 of Example 24 in tetrahydrofuran (7 mL) was added pyridine. (2.03 mL, 38.6 mmol) was added, tetradecanoyl chloride (0.930 mL, 3.77 mmol) was added under ice cooling, and the mixture was stirred at 60 ° C. for 2 hr. The reaction mixture was cooled to room temperature, water was added, and the mixture was extracted with ethyl acetate. The organic layer was washed with saturated brine, dried over anhydrous magnesium sulfate, and filtered. Concentration under reduced pressure, and the resulting residue was purified by silica gel column chromatography (hexane / ethyl acetate = 95/5 to 70/30) to give 2-((dimethylamino) methyl) -2- (hydroxymethyl) Propane-1,3-diyl ditetradecanoate (0.150 g, 0.257 mmol, yield 5%) was obtained.
ESI-MS m / z: 584 (M + H) ⁺
Process 2
Triethylamine was added to a solution of 2-((dimethylamino) methyl) -2- (hydroxymethyl) propane-1,3-diyl ditetradecanoate (0.060 g, 0.103 mmol) obtained in step 1 in dichloromethane (3 mL). (0.017 mL, 0.123 mmol) was added, and 4-nitrophenyl chloroformate (manufactured by Tokyo Chemical Industry Co., Ltd., 0.025 g, 0.123 mmol) and then tetradecylamine (manufactured by Tokyo Chemical Industry Co., Ltd., 0.022 g, 0.103 mmol) was added and stirred at room temperature overnight. Water was added to the reaction mixture, and the mixture was extracted with ethyl acetate. The organic layer was washed with saturated brine, dried over anhydrous magnesium sulfate, and filtered. The obtained residue was purified by silica gel column chromatography (hexane / ethyl acetate = 95/5 to 70/30) to give 2-((dimethylamino) methyl) -2-(((tetradecylcarbamoyl) oxy) Methyl) propane-1,3-diyl ditetradecanoate (0.052 g, 0.063 mmol, yield 62%) was obtained.
ESI-MS m / z: 824 (M + H) ⁺
Process 3
Using 2-((dimethylamino) methyl) -2-(((tetradecylcarbamoyl) oxy) methyl) propane-1,3-diyl ditetradecanoate (0.052 g, 0.063 mmol) obtained in Step 2 In the same manner as in Example 1, Step 2, the title compound (0.012 g, 0.014 mmol, yield 21%) was obtained.
ESI-MS m / z: 838 (M) ⁺ ; ¹ H-NMR (CDCl ₃ ) δ: 0.88 (t, J = 7.0 Hz, 9H), 1.22-1.33 (m, 62H), 1.51-1.61 (m, 6H), 2.38 (t, J = 7.6 Hz, 4H), 3.13 (dd, J = 14.2, 5.8 Hz, 2H), 3.59 (s, 9H), 4.12 (s, 2H), 4.19 (s, 2H), 4.21 (d, J = 12.0 Hz, 2H), 4.25 (d, J = 12.0 Hz, 2H), 6.72 (t, J = 5.8 Hz, 1H).

N, N, N-trimethyl-3-((octadecylcarbamoyl) oxy) -2,2-bis ((tetradecanoyloxy) methyl) propane-1-aminium chloride (Compound II-37)
Process 1
In the same manner as in Example 45, stearylamine (manufactured by Tokyo Chemical Industry Co., Ltd.) was used in place of tetradecylamine to give the title compound (0.015 g, 0.016 mmol, overall yield 0.5%).
ESI-MS m / z: 894 (M) +; 1 H-NMR (CDCl 3) δ: 0.88 (t, J = 6.8 Hz, 9H), 1.21-1.34 (m, 70H), 1.51-1.62 (m, 6H), 2.38 (t, J = 7.6 Hz, 4H), 3.13 (dd, J = 14.3, 5.8 Hz, 2H), 3.60 (s, 9H), 4.12 (s, 2H), 4.19 (s, 2H), 4.21 (d, J = 12.2 Hz, 2H), 4.25 (d, J = 12.2 Hz, 2H), 6.69 (t, J = 5.8 Hz, 1H).

N, N, N-trimethyl-3- (stearoyloxy) -2,2-bis ((tetradecanoyloxy) methyl) propane-1-aminium chloride (Compound II-37)
Process 1
(Angewandte Chemie International Edition), 2009, Vol. 48, p.2126-2130 (5- (Bromomethyl) -2,2-dimethyl-1,3- Stearoyl chloride (2.53 g, 8.36 mmol) was added to a solution of dioxane-5-yl) methanol (1.00 g, 4.18 mmol) in pyridine (10 mL), and the mixture was stirred at room temperature for 30 minutes. Water was added to the reaction mixture, and the mixture was extracted with ethyl acetate. The organic layer was washed with saturated brine, dried over anhydrous magnesium sulfate, and filtered. The obtained residue was purified by silica gel column chromatography (hexane / ethyl acetate = 90/10) to give (5- (bromomethyl) -2,2-dimethyl-1,3-dioxane-5-yl) methyl ester. An alert (0.95 g, 1.879 mmol, 45% yield) was obtained.
¹ H-NMR (CDCl ₃ ) δ: 0.89 (t, J = 7.0 Hz, 9H), 1.23-1.40 (m, 44H), 1.54-1.64 (m, 26H), 2.01-2.08 (m, 12H), 2.35 (t, J = 7.6 Hz, 6H), 2.77 (t, J = 6.8 Hz, 6H), 3.40 (s, 9H), 4.46 (s, 6H), 4.70 (s, 2H), 28-5.42 (m, 12H), 9.54 (br s, 1H).
Process 2
(5- (Bromomethyl) -2,2-dimethyl-1,3-dioxane-5-yl) methyl stearate (0.95 g, 1.879 mmol) N, N-dimethylformamide (10 mL) obtained in step 1 ) To the solution was added dimethylamine (2.0 mol / L tetrahydrofuran solution, 5.64 mL, 11.3 mmol), and the mixture was stirred at 120 ° C. for 13 hours under microwave irradiation. Water was added to the reaction mixture, and the mixture was extracted with ethyl acetate. The organic layer was washed with saturated brine, dried over anhydrous magnesium sulfate, and filtered. The obtained residue was purified by silica gel column chromatography (chloroform / methanol = 99/1 to 90/10) to give (5-((dimethylamino) methyl) -2,2-dimethyl-1,3-dioxane. -5-yl) methyl stearate (0.14 g, 0.298 mmol, 16% yield) was obtained.
ESI-MS m / z: 470 (M + H) ⁺
Process 3
(5-((Dimethylamino) methyl) -2,2-dimethyl-1,3-dioxane-5-yl) methyl stearate (0.140) obtained in Step 1 in the same manner as in Step 2 of Example 41. g, 0.298 mmol) was used to obtain 3- (dimethylamino) -2,2-bis (hydroxymethyl) propyl stearate (0.12 g, 0.279 mmol, yield 94%).
ESI-MS m / z: 430 (M + H) ⁺
Process 4
To a solution of 3- (dimethylamino) -2,2-bis (hydroxymethyl) propyl stearate (0.12 g, 0.279 mmol) obtained in step 3 in dichloromethane (2 mL), add pyridine (0.122 mL, 1.51 mmol). After that, tetradecanoyl chloride (0.224 g, 0.98 mmol) was added under ice cooling, and the mixture was stirred at room temperature for 1 hour. Water was added to the reaction mixture, and the mixture was extracted with ethyl acetate. The organic layer was washed with saturated brine, dried over anhydrous magnesium sulfate, and filtered. Concentrated under reduced pressure, and the resulting residue was purified by silica gel column chromatography (hexane / ethyl acetate = 95/5) to give 2-((dimethylamino) methyl) -2-((stearoyloxy) methyl) propane. -1,3-Diyl ditetradecanoate (0.150 g, 0.176 mmol, yield 63%) was obtained.
ESI-MS m / z: 851 (M + H) ⁺
Process 5
Using-((dimethylamino) methyl) -2-((stearoyloxy) methyl) propane-1,3-diyl ditetradecanoate (0.150 g, 0.176 mmol) obtained in step 4, Example 1 step In the same manner as in 2, the title compound (0.032 g, 0.056 mmol, yield 32%) was obtained.
ESI-MS m / z: 865 (M) ⁺ ; ¹ H-NMR (CDCl ₃ ) δ: 0.88 (t, J = 6.8 Hz, 9H), 1.19-1.33 (m, 68H), 1.54-1.65 (m, 6H), 2.38 (t, J = 7.6 Hz, 6H), 3.63 (s, 9H), 3.95 (s, 2H), 4.29 (s, 6H).

N, N, N-trimethyl-3-(((Z) -tetradec-9-enoyl) oxy) -2,2-bis ((((Z) -tetradec-9-enoyl) oxy) methyl) propane-1 -Aminium chloride (Compound II-39)
Process 1
To a solution of myristoleic acid (Nu-Chek Prep, Inc, 2.50 g, 11.0 mmol) in dichloromethane (20 mL) in thionyl chloride (1.61 mL, 22.1 mmol) and N, N-dimethylformamide (8.55 mL, 0.110 mmol) And stirred at room temperature for 1 hour. The reaction mixture was concentrated under reduced pressure to obtain a crude product of myristol chloride (2.70 g, 11.04 mmol, yield 100%).
Process 2
In the same manner as in Example 33, using myristolyl chloride (1.88 g, 7.66 mmol) obtained in Step 1 instead of palmitoyl chloride, the title compound (0.350 g, 0.417 mmol, overall yield 27%) was obtained. Obtained.
ESI-MS m / z: 803 (M) ⁺ ; ¹ H-NMR (CDCl ₃ ) δ: 0.86-0.94 (m, 9H), 1.26-1.39 (m, 36H), 1.53-1.64 (m, 6H), 1.97-2.07 (m, 12H), 2.38 (t, J = 7.6 Hz, 6H), 3.67 (s, 9H), 3.99 (s, 2H), 4.30 (s, 6H), 5.29-5.39 (m, 6H) .

2-((4-((1,3-bis (tetradecanoyloxy) -2-((tetradecanoyloxy) methyl) propan-2-yl) amino) -4-oxobutanoyl) oxy) -N , N, N-Trimethylethane-1-aminium chloride (Compound II-40)
Process 1
4-((1,3-Bis (tetra (tetrahydrochloride)) synthesized according to the method described in “Australian Journal of Chemistry”, 2013, Vol. 66, p.23-29. Decanoyloxy) -2-((tetradecanoyloxy) methyl) propan-2-yl) amino) -4-oxobutanoic acid (0.250 g, 0.293 mmol) in dichloromethane (3 mL) in 1-ethyl-3- (3-dimethylaminopropyl) carbodiimide (0.084 g, 0.440 mmol), 2- (dimethylamino) ethane-1-ol (manufactured by Tokyo Chemical Industry Co., Ltd., 0.039 g, 0.440 mmol), 4-dimethylaminopyridine (0.036 g, 0.293 mmol) was added in order and stirred at room temperature overnight. Water was added to the reaction mixture, and the mixture was extracted with ethyl acetate. The organic layer was washed with saturated brine, dried over anhydrous magnesium sulfate, and filtered. The residue obtained by concentration under reduced pressure was purified by silica gel column chromatography (chloroform / methanol = 99/1 to 90/10) to give 2- (4- (2- (dimethylamino) ethoxy) -4- Oxobutanamide) -2-((tetradecanoyloxy) methyl) propane-1,3-diyl ditetradecanoate (0.200 g, 0.217 mmol, 74% yield) was obtained.
ESI-MS m / z: 924 (M + H) ⁺
Process 2
2- (4- (2- (dimethylamino) ethoxy) -4-oxobutanamide) -2-((tetradecanoyloxy) methyl) propane-1,3-diyl ditetradecano obtained in step 1 The title compound (0.150 g, 0.154 mmol, 71% yield) was obtained in the same manner as in Example 1, Step 2 using art (0.200 g, 0.217 mmol).
ESI-MS m / z: 938 (M) ⁺ ; ¹ H-NMR (CDCl ₃ ) δ: 0.88 (t, J = 6.8 Hz, 9H), 1.20-1.34 (m, 60H), 1.56-1.65 (m, 6H), 2.34 (t, J = 7.6 Hz, 6H), 2.54 (br s, 4H), 3.48 (s, 9H), 4.13-4.21 (m, 2H), 4.40 (s, 6H), 4.57-4.65 ( m, 2H), 6.22 (s, 1H).

3-((4-((1,3-bis (tetradecanoyloxy) -2-((tetradecanoyloxy) methyl) propan-2-yl) amino) -4-oxobutanoyl) oxy) -1 -Methylquinuclidin-1-nium chloride (Compound II-41)
In the same manner as in Example 49, using kunuclidin-3-ol (manufactured by Tokyo Chemical Industry Co., Ltd.) instead of 2- (dimethylamino) ethane-1-ol, the title compound (0.350 g, 0.417 mmol, Rate 46%).
ESI-MS m / z: 976 (M) ⁺ ; ¹ H-NMR (CDCl ₃ ) δ: 0.88 (t, J = 7.0 Hz, 9H), 1.18-1.35 (m, 66H), 1.55-1.66 (m, 6H), 1.97-2.10 (m, 1H), 2.14-2.26 (m, 2H), 2.45-2.70 (m, 6H), 3.34 (s, 3H), 3.61-4.07 (m, 6H), 4.41 (s, 6H), 5.03-5.10 (m, 1H), 6.50 (s, 1H).

N, N, N-trimethyl-16,22-dioxo-19-(((tetradecylcarbamoyl) oxy) methyl) -17,21-dioxa-15,23-diazaheptatricontane-19-aminium chloride (compound II-42)
Process 1
2- (Dimethylamino) -2- (hydroxymethyl) propane-1,3-diol (0.15 g, 1.01 mmol) to toluene (4 mL), triethylamine (0.280 mL, 2.01 mmol), 1-tetradecane isocyanate (1.66 mL, 6.03 mmol) was added in order, and the mixture was reacted at 100 ° C. for 4 hours in a microwave reactor. Water was added to the reaction mixture, and the mixture was extracted with chloroform. The organic layer was washed with saturated brine, dried over anhydrous magnesium sulfate, and filtered. Concentration under reduced pressure, and the resulting residue was purified by silica gel column chromatography (chloroform / methanol = 99/1 to 90/10) to give 2- (dimethylamino) -2-(((tetradecylcarbamoyl) oxy ) Methyl) propane-1,3-diyl bis (tetradecylcarbamate) (0.872 g, 1.01 mmol, 100% yield).
ESI-MS m / z: 868 (M + H) ⁺
Process 2
Using 2- (dimethylamino) -2-(((tetradecylcarbamoyl) oxy) methyl) propane-1,3-diylbis (tetradecylcarbamate) (0.872 g, 1.01 mmol) obtained in step 1, In the same manner as in Example 1, step 2, the title compound (0.761 g, 0.829 mmol, yield 82%) was obtained.
ESI-MS m / z: 882 (M) ⁺ ; ¹ H-NMR (CDCl ₃ ) δ: 0.88 (t, J = 7.0 Hz, 9H), 1.22-1.32 (m, 66H), 1.47-1.56 (m, 6H), 3.13 (td, J = 14.3, 6.0 Hz, 6H), 3.58 (s, 9H), 4.52 (s, 6H), 6.69 (t, J = 6.0 Hz, 3H).

N, N, N-trimethyl-1,3-bis (3,7,11,15-tetramethylhexadecanoyloxy) -2-((3,7,11,15-tetramethylhexadecanoyl) methyl) Propan-2-aminium chloride (Compound II-43)
3,7,11,15-Tetramethylhexadecanoic acid (0.1826 g) in a solution of 2-dimethylamino-2-hydroxymethylpropane-1,3-diol (0.0170 g, 0.112 mmol) in 1,2-dichloroethane (1 mL) , 0.561 mmol), ((((1-cyano-2-ethoxy-2-oxoethylidene) amino) oxy) -4-morpholinomethylene) dimethylammonium hexafluorophosphate (Sigma-Aldirch, 0.240 g, 0.561 mmol), N, N-diisopropylethylamine (0.098 mL, 0.561 mmol) was added, and the mixture was stirred at 60 ° C. overnight. After cooling to room temperature, the mixture was concentrated under reduced pressure. The resulting residue was purified by amino silica gel column chromatography (hexane / ethyl acetate = 95/5) to give 2- (dimethylamino) -2-((3,7,11,15-tetramethylhexadecanoyloxy) A crude product of methyl) propane-1,3-diyldiyl bis (3,7,11,15-tetramethylhexadecanoate) was obtained. Methyl iodide (1.00 mL, 16.0 mmol) was added to the obtained crude product, and the mixture was stirred overnight at room temperature. The reaction solution was concentrated under reduced pressure. Dissolve the residue in a small amount of methanol-chloroform (1: 1) and ion exchange resin (Dow Chemical, Dowex (TM) 1x-2 100 mesh, Cl type, approx. Loaded and eluted with methanol-chloroform (1: 1). The eluate was concentrated under reduced pressure, and the resulting residue was purified by silica gel column chromatography (chloroform / methanol = 90/10) to obtain the title compound (0.0766 g, 0.071 mmol, yield 63%).
ESI-MS m / z: 1043 (M) ⁺ ; ¹ H-NMR (CDCl ₃ ) δ: 0.81-0.87 (m, 36H), 0.92 (d, J = 6.7 Hz, 9H), 0.97-1.42 (m, 60H), 1.51 (tt, J = 19.8, 6.7 Hz, 3H), 1.84-1.97 (m, 3H), 2.16 (ddd, J = 15.5, 8.4, 2.3 Hz, 3H), 2.38 (ddd, J = 15.5, 5.6, 1.6 Hz, 3H), 3.72 (s, 9H), 4.55 (s, 6H).

N, N, N-trimethyl-2-((tetradecanoyloxy) methyl) -2-tetradecylhexadecane-1-aminium chloride (Compound II-44)
(9Z, 12Z) -octadeca-9,12-dienyl methanesulfonate used in Step 1 of Example 19 and (9Z, 12Z) -octadeca-9,12-diene used in Step 4 in the same manner as Example 19. Instead of the acid, 1-bromotetradecane (manufactured by Tokyo Chemical Industry Co., Ltd.) and myristic acid (manufactured by Tokyo Chemical Industry Co., Ltd.) were used to obtain the title compound (0.292 g, 0.39 mmol, overall yield 22%).
ESI-MS m / z: 721 (M) ⁺ ; ¹ H-NMR (CD ₃ OD) δ: 0.90 (t, J = 6.8 Hz, 9H), 1.28-1.32 (m, 70H), 1.49 (br s, 2H), 1.63-1.66 (m, 2H), 2.42 (t, J = 7.2 Hz, 2H), 3.26 (s, 9H), 3.43 (s, 2H), 4.18 (s, 2H).

2-Hexadecyl-N, N, N-trimethyl-2-((palmitoyloxy) methyl) octadecane-1-aminium chloride (Compound II-45)
(9Z, 12Z) -octadeca-9,12-dienyl methanesulfonate used in Step 1 of Example 19 and (9Z, 12Z) -octadeca-9,12-diene used in Step 4 in the same manner as Example 19. Instead of the acid, 1-bromohexadecane (manufactured by Tokyo Chemical Industry Co., Ltd.) and palmitic acid (manufactured by Tokyo Chemical Industry Co., Ltd.) were used to obtain the title compound (0.195 g, 0.23 mmol, overall yield 5%).
ESI-MS m / z: 805 (M) ⁺ ; ¹ H-NMR (CD ₃ OD) δ: 0.90 (t, J = 6.8 Hz, 9H), 1.28-1.33 (m, 82H), 1.49 (br s, 2H), 1.63-1.67 (m, 2H), 2.43 (t, J = 7.2 Hz, 2H), 3.26 (s, 9H), 3.43 (s, 2H), 4.18 (s, 2H).

N, N, N-trimethyl-2-((stearoyloxy) methyl) -2-tetradecylhexadec-1-aminium chloride (Compound II-46)
(9Z, 12Z) -octadeca-9,12-dienyl methanesulfonate used in Step 1 of Example 19 and (9Z, 12Z) -octadeca-9,12-diene used in Step 4 in the same manner as Example 19. Instead of the acid, 1-bromotetradecane (manufactured by Tokyo Chemical Industry Co., Ltd.) and stearic acid (manufactured by Tokyo Chemical Industry Co., Ltd.) were used to obtain the title compound (0.421 g, 0.52 mmol, overall yield 20%).
ESI-MS m / z: 777 (M) ⁺ ; ¹ H-NMR (CD ₃ OD) δ: 0.90 (t, J = 6.8 Hz, 9H), 1.29-1.33 (m, 78H), 1.49 (br s, 2H), 1.63-1.67 (m, 2H), 2.43 (t, J = 7.2 Hz, 2H), 3.27 (s, 9H), 3.44 (s, 2H), 4.18 (s, 2H).

3- (Dodecanoyloxy) -N, N, N-trimethyl-2,2-bis ((stearoyloxy) methyl) propane-1-aminium chloride (Compound II-47)
In the same manner as in Example 46, instead of stearoyl chloride used in Step 46 of Example 46 and tetradecanoyl chloride used in Step 4, lauroyl chloride (manufactured by Tokyo Chemical Industry Co., Ltd.) and stearoyl chloride (manufactured by Tokyo Chemical Industry Co., Ltd.) were used. ) To give the title compound (0.200 g, 0.417 mmol, overall yield 0.3%).
ESI-MS m / z: 893 (M) ⁺ ; ¹ H-NMR (CDCl ₃ ) δ: 0.88 (t, J = 6.9 Hz, 9H), 1.21-1.32 (m, 72H), 1.57-1.64 (m, 6H), 2.38 (t, J = 7.6 Hz, 6H), 3.63 (s, 9H), 3.99 (s, 2H), 4.29 (s, 6H).

3- (Dodecanoyloxy) -N, N, N-trimethyl-2,2-bis ((palmitoyloxy) methyl) propane-1-aminium chloride (Compound II-48)
In the same manner as in Example 47, instead of stearoyl chloride used in Step 47 of Example 47 and tetradecanoyl chloride used in Step 4, lauroyl chloride (manufactured by Tokyo Chemical Industry Co., Ltd.) and palmitoyl chloride (Wako Pure Chemical Industries, Ltd.) were used. The title compound (0.350 g, 0.40 mmol, overall yield 0.6%) was obtained.
ESI-MS m / z: 837 (M) ⁺ ; ¹ H-NMR (CDCl ₃ ) δ: 0.88 (t, J = 6.9 Hz, 9H), 1.21-1.33 (m, 64H), 1.56-1.64 (m, 6H), 2.38 (t, J = 7.6 Hz, 6H), 3.66 (s, 9H), 3.98 (s, 2H), 4.29 (s, 6H).

3- (Dodecanoyloxy) -2-((dodecanoyloxy) methyl) -N, N, N-trimethyl-2-((stearoyloxy) methyl) propane-1-aminium chloride (Compound II-49)
In the same manner as in Example 47, substituting lauroyl chloride (manufactured by Tokyo Chemical Industry Co., Ltd.) for the tetradecanoyl chloride used in Step 47 of Example 47, the title compound (0.210 g, 0.249 mmol, overall yield 0.3%) )
ESI-MS m / z: 809 (M) ⁺ ; ¹ H-NMR (CDCl ₃ ) δ: 0.88 (t, J = 6.9 Hz, 9H), 1.23-1.34 (m, 60H), 1.53-1.65 (m, 6H), 2.38 (t, J = 7.6 Hz, 6H), 3.63 (s, 9H), 3.97 (s, 2H), 4.29 (s, 6H).

N, N, N-trimethyl-3- (palmitoyloxy) -2-((palmitoyloxy) methyl) -2-((stearoyloxy) methyl) propane-1-aminium chloride (compound II-50)
In the same manner as in Example 47, instead of tetradecanoyl chloride used in Step 4 of Example 47, palmitoyl chloride (manufactured by Tokyo Chemical Industry Co., Ltd.) was used, and the title compound (0.420 g, 0.44 mmol, overall yield 0.5) %).
ESI-MS m / z: 921 (M) ⁺ ; ¹ H-NMR (CDCl ₃ ) δ: 0.88 (t, J = 6.9 Hz, 9H), 1.21-1.32 (m, 76H), 1.55-1.65 (m, 6H), 2.38 (t, J = 7.6 Hz, 6H), 3.67 (s, 9H), 399 (s, 2H), 4.29 (s, 6H).

3- (Icosanoyloxy) -N, N, N-trimethyl-2,2-bis ((tetradecanoyloxy) methyl) propane-1-aminium chloride (Compound II-51)
In the same manner as in Example 47, using eicosanoyl chloride (Nu-Chek Prep, Inc.) instead of stearoyl chloride used in Step 47 of Example 47, the title compound is obtained.

3- (Dodecanoyloxy) -2-((dodecanoyloxy) methyl) -2-((icosanoyloxy) methyl) -N, N, N-trimethylpropane-1-aminium chloride (Compound II-52)
In the same manner as in Example 47, instead of stearoyl chloride used in Step 47 of Example 47 and tetradecanoyl chloride used in Step 4, eicosanoyl chloride (manufactured by Nu-Chek Prep, Inc.) and lauroyl chloride (Japanese The title compound is obtained using Kogaku Pharmaceutical Co., Ltd.

N, N, N-trimethyl-3- (methyl (3- (tetradecanoyloxy) -2,2-bis ((tetradecanoyloxy) methyl) propyl) amino) propane-1-aminium chloride (compound II- 53)
Process 1
To a solution of 2- (bromomethyl) -2- (hydroxymethyl) propane-1,3-diol (0.15 g, 0.754 mmol) in N, N-dimethylacetamide (1 mL), N, N, N'-trimethylpropane- 1,3-diamine (0.263 g, 2.26 mmol) was added, and the mixture was reacted at 100 ° C. for 2 hours in a microwave reactor. Then, N, N-diisopropylethylamine (0.395 mL, 2.26 mmol) and then tetradecanoyl chloride (1.12 g, 4.52 mmol) were added under ice cooling, and the mixture was stirred at room temperature for 2 hours. Water was added to the reaction mixture, and the mixture was extracted with ethyl acetate. The organic layer was washed with saturated aqueous sodium hydrogen carbonate solution and saturated brine, dried over anhydrous magnesium sulfate, and filtered. Concentration under reduced pressure, and the resulting residue was purified by silica gel column chromatography (chloroform / methanol = 99/1 to 90/10) to give 2-((((3- (dimethylamino) propyl) (methyl) amino. ) Methyl) -2-((tetradecanoyloxy) methyl) propane-1,3-diyl ditetradecanoate (0.040 g, 0.046 mmol, 6% yield) was obtained.
ESI-MS m / z: 866 (M + H) ⁺
Process 2
2-((((3- (dimethylamino) propyl) (methyl) amino) methyl) -2-((tetradecanoyloxy) methyl) propane-1,3-diyl ditetradecanoate obtained in step 1 (0.040 g, 0.046 mmol) was used in the same manner as in Step 1 of Example 1 to obtain the title compound (0.015 g, 0.016 mmol, yield 35%).
ESI-MS m / z: 880 (M) ⁺ ; ¹ H-NMR (CDCl ₃ ) δ: 0.88 (t, J = 6.8 Hz, 9H), 1.20-1.35 (m, 62H), 1.54-1.65 (m, 6H), 1.87-2.03 (m, 2H), 2.21-2.31 (m, 2H), 2.31 (t, J = 7.6 Hz, 6H), 2.49 (br s, 3H), 3.40 (s, 9H), 3.52- 3.63 (m, 2H), 4.05 (s, 6H).

(S) -6- (Di (9Z, 12Z) -octadeca-9,12-dienylamino) -N, N, N-trimethyl-5-oleamido-6-oxohexane-1-aminium chloride (Compound III-2)
Process 1
Add (9Z, 12Z) -octadeca-9,12-dienyl methanesulfonate (1.55 g, 4.50 mmol) to ammonia (approximately 2 mol / L methanol solution, 18.0 mL, 36.0 mmol) and use a microwave reactor. And stirred at 130 ° C. for 3 hours. A saturated aqueous sodium hydrogen carbonate solution was added to the reaction mixture, and the mixture was extracted 5 times with chloroform. The organic layers were combined, washed with saturated brine, dried over anhydrous magnesium sulfate, filtered, and concentrated under reduced pressure to obtain a crude product of (Z) -octadeca-9-enylamine.
Add (9Z, 12Z) -octadeca-9,12-dienyl methanesulfonate (1.24 g, 3.60 mmol) and 50% aqueous sodium hydroxide (1.44 g, 18.0 mmol) to the resulting crude product, and add to the oil bath. The mixture was stirred at 110 ° C. for 60 minutes. After cooling to room temperature, the reaction mixture was diluted with ethyl acetate, washed with water and then with saturated brine, dried over anhydrous magnesium sulfate, filtered, and concentrated under reduced pressure. The obtained residue was purified by silica gel column chromatography (chloroform / methanol = 100/0 to 95/5) to give (9Z, 12Z) -di (9Z, 12Z) -octadeca-9,12-dienylamine (0.838 g, 1.631 mmol, yield 36%).
ESI-MS m / z: 515 (M + H) +; 1H-NMR (CDCl3) δ: 0.89 (t, J = 6.8 Hz, 6H), 1.26-1.38 (m, 32H), 1.45-1.54 (m, 4H), 2.05 (q, J = 6.6 Hz, 8H), 2.60 (t, J = 7.1 Hz, 4H), 2.77 (t, J = 5.9 Hz, 4H), 5.29-5.43 (m, 8H).
Process 2
To a solution of (S) -2-amino-6- (tert-butoxycarbonylamino) hexanoic acid (1.94 g, 7.88 mmol) in acetone (5 mL), sodium hydroxide (2 mol / L aqueous solution, 5 mL), Oil chloride (2.09 g, 6.89 mmol) was added, and the mixture was stirred at room temperature overnight. A hydrochloric acid aqueous solution (6 mol / L) was added to the reaction solution, and the mixture was extracted twice with chloroform. The organic layer was dried over anhydrous magnesium sulfate and concentrated under reduced pressure. The residue was purified by silica gel column chromatography (chloroform / methanol = 90 / 10-80 / 20) to give (S) -6- (tert-butoxycarbonylamino) -2-oleamidohexanoic acid (2.50 g, 4.89 mmol Yield 71%).
ESI-MS m / z: 510 (M-H) ^- ; ¹ H-NMR (CDCl3) δ: 0.87 (t, J = 7.0 Hz, 3H), 1.20-1.54 (m, 33H), 1.57-1.68 (m , 2H), 1.71-1.93 (m, 2H), 1.96-2.05 (m, 4H), 2.18-2.29 (m, 2H), 3.07-3.16 (m, 2H), 4.50-4.60 (m, 1H), 4.63 -4.76 (m, 1H), 5.28-5.39 (m, 2H), 6.49-6.57 (m, 1H).
Process 3
To a solution of (S) -6- (tert-butoxycarbonylamino) -2-oleamidohexanoic acid (0.291 g, 0.570 mmol) obtained in step 2 in 1,2-dichloroethane (4 mL), O- (7 -Aza-1H-benzotriazol-1-yl) -N, N, N ', N'-tetramethyluronium hexafluorophosphate (0.433 g, 1.14 mmol), N, N-diisopropylethylamine (0.498 mL, 2.85 mmol), (9Z, 12Z) -di (9Z, 12Z) -octadeca-9,12-dienylamine (0.293 g, 0.570 mmol) obtained in Step 1 was added and stirred at room temperature for 4 hours. A saturated aqueous sodium hydrogen carbonate solution was added to the reaction mixture, and the mixture was extracted twice with chloroform. The organic layer was dried over anhydrous magnesium sulfate, filtered and concentrated under reduced pressure. The residue was purified by silica gel column chromatography (hexane / ethyl acetate = 80/20) to give tert-butyl (S) -6- (di (9Z, 12Z) -octadeca-9,12-dienylamino) -5-oleamide -6-oxohexyl carbamate (0.489 g, 0.486 mmol, 85% yield) was obtained.
ESI-MS m / z: 1008 (M + H) ⁺ ; ¹ H-NMR (CDCl3) δ: 0.85-0.92 (m, 9H), 1.20-1.72 (m, 73H), 1.97-2.08 (m, 12H) , 2.18 (t, J = 7.6 Hz, 2H), 2.74-2.80 (m, 4H), 3.02-3.34 (m, 5H), 3.44-3.53 (m, 1H), 4.55-4.63 (m, 1H), 4.88 (td, J = 8.2, 4.6 Hz, 1H), 5.28-5.43 (m, 10H), 6.30 (d, J = 8.4 Hz, 1H).
Process 4
Of tert-butyl (S) -6- (di (9Z, 12Z) -octadeca-9,12-dienylamino) -5-oleamido-6-oxohexylcarbamate (0.459 g, 0.456 mmol) obtained in step 3 Trifluoroacetic acid (0.500 mL, 6.49 mmol) was added to the 1,2-dichloroethane (2 mL) solution, and the mixture was stirred at room temperature for 1 hour. The reaction mixture was concentrated under reduced pressure, chloroform and saturated aqueous sodium hydrogen carbonate solution were added to the residue, and the mixture was extracted twice with chloroform. The organic layer was dried over anhydrous magnesium sulfate, filtered and concentrated under reduced pressure. The residue was purified by silica gel column chromatography (hexane / ethyl acetate = 90 / 10-80 / 20) to give N-((S) -6-amino-1- (di ((9Z, 12Z) -octadeca-9 , 12-Dien-1-yl) amino) -1-oxohexane-2-yl) oleamide (0.259 g, 0.286 mmol, yield 63%) was obtained.
ESI-MS m / z: 907 (M + H) ⁺ ; ¹ H-NMR (CDCl3) δ: 0.86-0.91 (m, 9H), 1.20-1.71 (m, 64H), 1.96-2.09 (m, 12H) , 2.21 (t, J = 7.5 Hz, 2H), 2.73-2.88 (m, 6H), 3.08-3.47 (m, 4H), 4.81-4.88 (m, 1H), 5.28-5.43 (m, 10H), 6.67 (br s, 1H).
Process 5
N-((S) -6-amino-1- (di ((9Z, 12Z) -octadec-9,12-dien-1-yl) amino) -1-oxohexane-2-obtained in step 4 Yl) oleamide (0.137 g, 0.151 mmol) in 1,2-dichloroethane (1 mL) was added 38% aqueous formaldehyde solution (0.300 mL) and sodium triacetoxyborohydride (0.096 g, 0.453 mmol), and at room temperature overnight. Stir. A saturated aqueous sodium hydrogen carbonate solution was added to the reaction mixture, and the mixture was extracted twice with chloroform. The organic layer was dried over anhydrous magnesium sulfate and concentrated under reduced pressure. The residue was purified by amino silica gel column chromatography (hexane / ethyl acetate = 50/50) to give N-((S) -1- (di (9Z, 12Z) -octadeca-9,12-dienylamino) -6. -(Dimethylamino) -1-oxohexane-2-yl) oleamide (0.122 g, 0.130 mmol, yield 86%) was obtained.
ESI-MS m / z: 936 (M + H) ⁺ ; ¹ H-NMR (CDCl ₃ ) δ: 0.83-0.93 (m, 9H), 1.12-1.77 (m, 64H), 1.95-2.25 (m, 22H ), 2.73-2.80 (m, 4H), 3.04-3.15 (m, 1H), 3.20-3.34 (m, 2H), 3.44-3.54 (m, 1H), 4.85-4.91 (m, 1H), 5.28-5.43 (m, 10H), 6.28 (d, J = 8.6 Hz, 1H).
Process 6
Example 8 In the same manner as in Step 2, (9Z, 9'Z, 12Z, 12'Z) -2- (dimethylamino) -2-(((9Z, 12Z) -octadec-9,12-dienoyloxy) N-((S) -1- (di (9Z, 12Z) -octadeca-9,12-dienylamino) obtained in step 5 instead of (methyl) propane-1,3-diyl dioctadec-9,12-dienoate The title compound (0.0707 g, 0.0718 mol, 65%) was obtained using -6- (dimethylamino) -1-oxohexane-2-yl) oleamide (0.104 g, 0.111 mol).
ESI-MS m / z: 950 (M) ⁺ ; ¹ H-NMR (CDCl ₃ ) δ: 0.85-0.91 (m, 9H), 1.21-1.83 (m, 64H), 1.97-2.08 (m, 12H), 2.19 (t, J = 7.7 Hz, 2H), 2.74-2.80 (m, 4H), 3.05-3.84 (m, 15H), 4.82-4.90 (m, 1H), 5.28-5.43 (m, 10H), 6.41- 6.46 (m, 1H).

(S) -N, N, N-Trimethyl-5- (nonacosan-15-yloxy) -1,5-dioxo-1- (tetradecyloxy) pentane-2-aminium chloride (Compound III-3)
Process 1
Tetradecylmagnesium chloride (Sigma-Aldrich, 1.0 mol / L tetrahydrofuran solution, 59.4 mL, 59.4 mmol) was added to a solution of ethyl formate (Nacalai Tesque, 2.4 mL, 29.7 mmol) in tetrahydrofuran (9 mL). The mixture was stirred at 60 ° C. for 2 hours. The reaction solution was ice-cooled, and water and sulfuric acid (Nacalai Tesque, 2.0 mol / L aqueous solution) were added. The precipitate was collected by filtration to obtain nonacosan-15-ol (6.90 g, 16.2 mmol, yield 55%).
¹ H-NMR (CDCl ₃ ) δ: 0.88 (t, J = 6.8 Hz, 6H), 1.22-1.34 (m, 48H), 1.37-1.49 (m, 2H), 3.54-3.64 (m, 1H).
Process 2
Paraformaldehyde (Sigma-Aldrich, 5.50 g, 183 mmol) in a solution of 1-tert-butyl 2-aminopentanedioate hydrochloride (Watanabe Chemical Co., Ltd., 10.0 g, 30.3 mmol) in ethanol (150 mL), Sodium cyanoborohydride (5.70 g, 90.7 mmol) was added, and the mixture was stirred overnight at room temperature. A saturated aqueous sodium hydrogen carbonate solution was added to the reaction mixture, and the mixture was extracted twice with chloroform. The organic layer was washed with saturated brine, dried over anhydrous magnesium sulfate, filtered, and concentrated under reduced pressure. The residue was purified by silica gel column chromatography (hexane / ethyl acetate = 35/65) to give (S) -5-benzyl 1-tert-butyl 2- (dimethylamino) pentanedioate (8.20 g, 25.5 mmol, Yield 84%) was obtained.
¹ H-NMR (CDCl ₃ ) δ: 1.48 (s, 9H), 1.95-2.00 (m, 2H), 2.32 (s, 6H), 2.43 (t, J = 7.8 Hz, 2H), 3.04 (t, J = 7.5 Hz, 1H), 5.12 (s, 6H), 7.29-7.40 (m, 5H).
Process 3
To a solution of (S) -5-benzyl 1-tert-butyl 2- (dimethylamino) pentanedioate (8.20 g, 25.5 mmol) in ethanol (200 mL), palladium-carbon (Tokyo Chemical Industry Co., Ltd., 10% palladium) , About 55% wet water product, 820 mg), and stirred for 7 hours at room temperature under hydrogen atmosphere. Insolubles were removed by celite filtration, and the filtrate was concentrated. The resulting residue was purified by silica gel column chromatography (dichloromethane / methanol = 85/15) to give a crude product of (S) -5-tert-butoxy-4- (dimethylamino) -5-oxopentanoic acid Got. (4.83 g, 20.9 mmol, crude yield 82%) was obtained.
To the obtained crude product of (S) -5-tert-butoxy-4- (dimethylamino) -5-oxopentanoic acid (4.83 g, 20.9 mmol), 1.2-dichloroethane (200 mL) was obtained in Step 1. Nonacosan-15-ol (9.75 g, 23.0 mmol), 1-ethyl-3- (3-dimethylaminopropyl) carbodiimide hydrochloride (8.01 g, 41.8 mmol), N, N-dimethylaminopyridine (255 mg, 2.09 mmol) was added and the mixture was stirred at 50 ° C. for 3 hours. Water was added to the reaction mixture, and the mixture was extracted twice with dichloromethane. The organic layer was washed with saturated brine, dried over anhydrous magnesium sulfate, filtered, and concentrated under reduced pressure. The resulting residue was purified by silica gel column chromatography (hexane / ethyl acetate = 85/15) to give (S) -1-tert-butyl 5-nonacosan-15-yl 2- (dimethylamino) pentanedioate (8.13 g, 12.7 mmol, 61% yield) was obtained.
¹ H-NMR (CDCl ₃ ) δ: 0.88 (t, J = 6.8 Hz, 6H), 1.20-1.38 (m, 48H), 1.46-1.57 (m, 4H), 1.91-2.00 (m, 2H), 2.33 -2.41 (m, 8H), 3.05 (t, J = 7.6 Hz, 1H), 4.82-4.93 (m, 1H).
Process 4
To a solution of (S) -1-tert-butyl 5-nonacosan-15-yl 2- (dimethylamino) pentanedioate (8.13 g, 12.7 mmol) obtained in step 3 in dichloromethane (40 mL), trifluoroacetic acid was added. (20 mL) was added, and the mixture was stirred at 40 ° C. overnight. The reaction solution was concentrated under reduced pressure. The resulting residue was purified by silica gel column chromatography (dichloromethane / methanol = 85/15) to obtain (S) -2- (dimethylamino) -5- (nonacosan-15-yloxy) -5-oxopentanoic acid (6.70 g, 11.5 mmol, 90% yield) was obtained.
¹ H-NMR (CDCl ₃ ) δ: 0.89 (t, J = 7.0 Hz, 6H), 1.18-1.37 (m, 48H), 1.44-1.60 (m, 4H), 2.00-2.13 (m, 2H), 2.52 -2.74 (m, 2H), 2.87 (s, 6H), 3.62-3.73 (m, 1H), 4.80-4.89 (m, 1H).
Process 5
1,2-Dichloroethane solution (2.0 mL) of (S) -2- (dimethylamino) -5- (nonacosan-15-yloxy) -5-oxopentanoic acid (100 mg, 0.172 mmol) obtained in step 4 (1-cyano-2-ethoxy-2-oxoethylideneaminooxy) dimethylamino-morpholino-carbenium hexafluorophosphate (96.0 mg, 0.224 mmol), N, N-diisopropylethylamine (0.060 mL, 0.344 mmol) ) Was added tetradecan-1-ol () and stirred at room temperature overnight. Water was added to the reaction mixture, and the mixture was extracted with dichloromethane. The organic layer was washed with water, dried over anhydrous magnesium sulfate, filtered and concentrated under reduced pressure. The resulting residue was purified by silica gel column chromatography (hexane / ethyl acetate = 95/5 to 85/15) to give (S) -5-nonacosan-15-yl 1-tetradecyl 2- (dimethylamino) pentane Geoart (64.0 mg, 0.0822 mmol, 48% yield) was obtained. ¹ H-NMR (CDCl ₃ ) δ: 0.88 (t, J = 6.8 Hz, 9H), 1.12-1.39 (m, 70H), 1.45-1.69 (m, 6H), 1.93-2.03 (m, 2H), 2.29 -2.38 (m, 8H), 3.16 (t, J = 7.4 Hz, 1H), 4.03-4.17 (m, 2H), 4.80-4.91 (m, 1H).
Process 6
Example 8 In the same manner as in Step 2, (9Z, 9'Z, 12Z, 12'Z) -2- (dimethylamino) -2-(((9Z, 12Z) -octadec-9,12-dienoyloxy) (S) -5-Nonacosan-15-yl 1-tetradecyl 2- (dimethylamino) pentanedioate obtained in step 5 instead of methyl) propane-1,3-diyl dioctadec-9,12-dienoate The title compound is obtained.

(S) -1- (Dodecyloxy) -N, N, N-trimethyl-5- (nonacosan-15-yloxy) -1,5-dioxopentane-2-aminium chloride (Compound III-4)
In a manner similar to Example 64, substituting dodecan-1-ol for tetradecan-1-ol in Example 64, step 5, to give the title compound.

(S) -1- (Hexadecyloxy) -N, N, N-trimethyl-5- (nonacosan-15-yloxy) -1,5-dioxopentane-2-aminium chloride (Compound III-5)
In the same manner as in Example 64, substituting hexadecan-1-ol for tetradecan-1-ol in Step 64 of Example 64 to give the title compound.

(S) -N, N, N-Trimethyl-5- (nonacosan-15-yloxy) -1- (octadecyloxy) -1,5-dioxopentane-2-aminium chloride (Compound III-6)
In a manner similar to Example 64, substituting octadecan-1-ol for tetradecan-1-ol in Step 5 of Example 64 to give the title compound.

(S, Z) -N, N, N-Triethyl-5- (nonacosan-15-yloxy) -1- (octadeca-9-enyloxy) -1,5-dioxopentane-2-aminium chloride (Compound III- 7)
In the same manner as in Example 64, substituting (Z) -octadec-9-en-1-ol for tetradecan-1-ol in Step 5 of Example 64 to give the title compound.

(6Z, 9Z, 28Z, 31Z) -N, N-Dimethyl-N- (2- (N-methylstearamido) ethyl) heptatriconta-6,9,28,31-tetraene-19-aminium chloride ( Compound IV-2)
Process 1
(6Z, 9Z, 28Z, 31Z) -heptatriconta-6,9,28,31-tetraen-19-one (0.50 g, obtained by a method according to the method described in WO2010 / 042877 0.256 mmol) in 1,2-dichloroethane (2 mL), methanol (2 mL), N1, N2-dimethylethane-1,2-diamine (manufactured by Tokyo Chemical Industry Co., Ltd., 0.085 mL, 0.767 mmol), Acetoxyborohydride (0.325 g, 1.53 mmol) was added and stirred at 50 ° C. for 5 hours. A saturated aqueous sodium hydrogen carbonate solution was added to the reaction mixture, and the mixture was extracted twice with chloroform. The organic layer was dried over anhydrous magnesium sulfate, filtered and concentrated under reduced pressure. The resulting residue was purified by amino silica column chromatography (hexane / ethyl acetate = 90 / 10-80 / 20) to give N1-((6Z, 9Z, 28Z, 31Z) -heptatria contour-6,9,28 , 31-tetraen-19-yl) -N1, N2-dimethylethane-1,2-diamine (0.0303 g, 0.0506 mmol, yield 20%) was obtained.
ESI-MS m / z: 600 (M + H) ⁺ ; ¹ H-NMR (CDCl ₃ ) δ: 0.89 (t, J = 6.8 Hz, 6H), 1.13-1.45 (m, 40H), 2.01-2.09 ( m, 8H), 2.14 (s, 3H), 2.28-2.40 (m, 1H), 2.43 (s, 3H), 2.52-2.60 (m, 4H), 2.75-2.80 (m, 4H), 5.29-5.42 ( m, 8H).
Process 2
N1-((6Z, 9Z, 28Z, 31Z) -heptatriconta-6,9,28,31-tetraen-19-yl) -N1, N2-dimethylethane-1,2-diamine obtained in Step 1 To a solution of (0.0258 g, 0.0431 mmol) in 1,2-dichloroethane (1 mL), stearoyl chloride (0.039 0 g, 0.129 mmol) and N, N-diisopropylethylamine (0.038 mL, 0.215 mmol) were added, and 1 at room temperature. Stir for hours. A saturated aqueous sodium hydrogen carbonate solution was added to the reaction mixture, and the mixture was extracted twice with chloroform. The organic layer was dried over anhydrous magnesium sulfate, filtered and concentrated under reduced pressure. The obtained residue was purified by aminosilica column chromatography (hexane / ethyl acetate = 90/10) A crude product of 31-tetraen-19-yl (methyl) amino) ethyl) -N-methylstearamide was obtained.
Methyl iodide (1.00 mL, 16.0 mmol) was added to the obtained crude product, and the mixture was stirred at 50 ° C. for 1 hour. The reaction solution was concentrated under reduced pressure. Dissolve the residue in a small amount of methanol-chloroform (1: 1) and use it as an ion exchange resin (Dow Chemical, Dowex (TM) 1x-2 100 mesh, Cl type, approximately 20 times the volume, pre-washed with water and methanol). Loaded and eluted with methanol-chloroform (1: 1). The eluate was concentrated under reduced pressure, and the resulting residue was purified by silica gel column chromatography (chloroform / methanol = 80/20) to obtain the title compound (0.0149 g, 0.0163 mmol, yield 38%).
ESI-MS m / z: 881 (M) ⁺ ; ¹ H-NMR (CDCl ₃ ) δ: 0.84-0.92 (m, 9H), 1.20-2.09 (m, 78H), 2.27-2.38 (m, 2H), 2.74-2.80 (m, 4H), 3.14-3.45 (m, 9H), 3.77-4.09 (m, 5H), 5.28-5.43 (m, 8H).

(9Z, 12Z) -N, N-Dimethyl-N- (3-((9Z, 12Z) -N-((9Z, 12Z) -octadeca-9,12-dienyl) octadeca-9,12-dienamido) propyl ) Octadeca-9,12-diene-1-aminium chloride (Compound IV-3)
Process 1
Add 3-aminopropan-1-ol (1.66 g, 21.9 mmol) to (9Z, 12Z) -octadeca-9,12-dienyl methanesulfonate (0.838 g, 2.43 mmol) and stir at 90 ° C for 3 hours did. The reaction mixture was concentrated under reduced pressure, and the resulting residue was purified by amino silica gel chromatography (hexane / ethyl acetate) to give 3-((9Z, 12Z) -octadeca-9,12-dienylamino) propan-1-ol ( 0.722 g, 2.23 mmol, 92% yield).
ESI-MS m / z: 325 (M + H) ⁺ ; ¹ H-NMR (CDCl ₃ ) δ: 0.89 (t, J = 6.9 Hz, 3H), 1.26-1.39 (m, 17H), 1.46 (tt, J = 7.1, 6.9 Hz, 3H), 1.69 (tt, J = 5.7, 5.4 Hz, 2H), 2.02-2.08 (m, 4H), 2.60 (t, J = 7.1 Hz, 2H), 2.75-2.80 (m , 2H), 2.88 (t, J = 5.7 Hz, 2H), 3.81 (t, J = 5.4 Hz, 2H), 5.30-5.42 (m, 4H).
Process 2
Example 63 N-((S) -6-amino-1- (di ((9Z, 12Z) -octadec-9,12-dien-1-yl) amino) -1- Instead of oxohexane-2-yl) oleamide, 3-((9Z, 12Z) -octadeca-9,12-dienylamino) propan-1-ol (0.233 g, 0.722 mol) obtained in Step 1 was used. -(Methyl ((9Z, 12Z) -octadeca-9,12-dienyl) amino) propan-1-ol (0.220 g, 0.652 mol, yield 90%) was obtained.
ESI-MS m / z: 338 (M + H) ⁺ ; ¹ H-NMR (CDCl ₃ ) δ: ¹ H-NMR (CDCl ₃ ) δ: 0.89 (t, J = 7.0 Hz, 3H), 1.24-1.40 (m, 16H), 1.47 (tt, J = 7.6, 7.0 Hz, 2H), 1.69 (tt, J = 5.7, 5.2 Hz, 2H), 2.01-2.08 (m, 4H), 2.23 (s, 3H), 2.34 (t, J = 7.6 Hz, 2H), 2.59 (t, J = 5.7 Hz, 2H), 2.75-2.80 (m, 2H), 3.80 (t, J = 5.2 Hz, 2H), 5.29-5.42 (m , 4H).
Process 3
To a solution of (9Z, 12Z) -octadeca-9,12-dienyl methanesulfonate (2.85 g, 8.27 mmol) in acetonitrile (30 mL), cesium carbonate (6.74 g, 20.7 mmol), tetra-n-butylammonium iodide (3.05 g, 8.27 mmol) and N- (tert-butoxycarbonyl) -2-nitrobenzenesulfonamide (2.50 g, 8.27 mmol) were added, and the mixture was stirred for 3 hours with heating under reflux. The reaction mixture was cooled to room temperature, water was added, and the mixture was extracted with ethyl acetate. The organic layer was dried over anhydrous magnesium sulfate, filtered and concentrated under reduced pressure. The resulting residue was purified by silica gel column chromatography (hexane / ethyl acetate = 70/30) to give tert-butyl 2-nitrophenylsulfonyl ((9Z, 12Z) -octadeca-9,12-dienyl) carbamate (3.21 g , 5.83 mmol).
To a solution of tert-butyl 2-nitrophenylsulfonyl ((9Z, 12Z) -octadeca-9,12-dienyl) carbamate (3.21 g, 5.83 mmol) in dichloromethane (23 mL), trifluoroacetic acid (9.63 mL, 126 mmol) was added and stirred at room temperature for 0.5 hour. The reaction solution was diluted with dichloromethane, and an aqueous sodium hydroxide solution (1 mol / L) and a saturated aqueous sodium hydrogen carbonate solution were added. The organic layer was washed with saturated brine, dried over anhydrous magnesium sulfate, filtered, and concentrated under reduced pressure. The obtained residue was purified by silica gel column chromatography (hexane / ethyl acetate = 95/5) to give 2-nitro-N-((9Z, 12Z) -octadeca-9,12-dienyl) benzenesulfonamide (2.48 g , 5.50 mmol, 67% yield).
ESI-MS m / z: 338 (M + H) ⁺ ; ¹ H-NMR (CDCl ₃ ) δ: 0.89 (t, J = 7.0 Hz, 3H), 1.22-1.39 (m, 16H), 1.52 (m, 2H), 2.01-2.05 (m, 4H), 2.77 (t, J = 6.6 Hz, 2H), 3.09 (q, J = 6.7 Hz, 2H), 5.23 (m, 1H), 5.31-5.42 (m, 4H ), 7.71-7.76 (m, 2H), 7.78-7.87 (1H), 813-8.15 (m, 1H).
Process 4
To a solution of 3- (methyl ((9Z, 12Z) -octadeca-9,12-dienyl) amino) propan-1-ol (0.220 g, 0.652 mol) obtained in step 2 in tetrahydrofuran (4 mL), add step 3 2-Nitro-N-((9Z, 12Z) -octadeca-9,12-dienyl) benzenesulfonamide (0.441 g, 0.978 mmol), triphenylphosphine (0.257 g, 0.978 mmol), azodicarboxylic acid Diethyl (Nacalai Tesque, 40% toluene solution, 0.387 mL, 0.851 mmol) was added, and the mixture was stirred at 50 ° C. for 2 hr. The reaction mixture was cooled to room temperature, saturated brine was added, and the mixture was extracted twice with hexane. The organic layer was dried over anhydrous magnesium sulfate, filtered and concentrated under reduced pressure. The resulting residue was purified by amino silica gel column chromatography (hexane / ethyl acetate = 80/20) to give N- (3- (methyl ((9Z, 12Z) -octadeca-9,12-dienyl) amino) propyl) A crude product of -2-nitro-N-((9Z, 12Z) -octadeca-9,12-dienyl) benzenesulfonamide was obtained.
N- (3- (methyl ((9Z, 12Z) -octadeca-9,12-dienyl) amino) propyl) -2-nitro-N-((9Z, 12Z) -octadeca-9,12-dienyl obtained ) To a solution of the crude product of benzenesulfonamide in acetonitrile (5 mL), dodecane-1-thiol (0.409 mL, 1.63 mmol) and 1,8-diazabicyclo [5.4.0] -7-undecene (0.246 mL, 1.630 mmol) ) Was added and stirred at 60 ° C. for 2 hours. Water was added to the reaction mixture, and the mixture was extracted twice with hexane. The organic layer was washed with saturated brine, dried over anhydrous magnesium sulfate, filtered, and concentrated under reduced pressure. The resulting residue was purified by amino silica gel column chromatography (hexane / ethyl acetate = 75/25) to give N1-methyl-N1, N3-di ((9Z, 12Z) -octadeca-9,12-dienyl) propane- 1,3-diamine (0.212 g, 0.363 mmol, yield 56%) was obtained.
ESI-MS m / z: 586 (M + H) ⁺ ; ¹ H-NMR (CDCl ₃ ) δ: 0.89 (t, J = 7.0 Hz, 6H), 1.22-1.51 (m, 36H), 1.66 (tt, J = 7.2, 7.1 Hz, 2H), 2.01-2.08 (m, 8H), 2.20 (s, 3H), 2.29 (t, J = 7.6 Hz, 2H), 2.36 (t, J = 7.2 Hz, 2H), 2.58 (t, J = 7.4 Hz, 2H), 2.62 (t, J = 7.1 Hz, 2H), 2.75-2.80 (m, 4H), 5.29-5.43 (m, 8H).
Process 5
N1-methyl-N1, N3-di ((9Z, 12Z) -octadeca-9,12-dienyl) propane-1,3-diamine (0.108 g, 0.185 mmol) of 1,2-dichloroethane obtained in step 4 (1 mL) To the solution, add (9Z, 12Z) -octadeca-9,12-dienoic acid (0.104 g, 0.370 mmol), 1-ethyl-3- (3-dimethylaminopropyl) carbodiimide hydrochloride (0.106 g, 0.555 mmol) and N, N-dimethylaminopyridine (0.0023 g, 0.0188 mmol) were added, and the mixture was stirred at room temperature for 1 hour. The reaction solution was concentrated under reduced pressure. The resulting residue was purified by amino silica gel column chromatography (hexane / ethyl acetate = 85/15) to give (9Z, 12Z) -N- (3- (methyl ((9Z, 12Z) -octadeca-9,12- Dienyl) amino) propyl) -N-((9Z, 12Z) -octadeca-9,12-dienyl) octadeca-9,12-dienamide (0.146 g, 0.172 mmol, 93% yield) was obtained.
ESI-MS m / z: 848 (M + H) ⁺ ; ¹ H-NMR (CDCl ₃ ) δ: 0.89 (t, J = 7.0 Hz, 9H), 1.21-1.74 (m, 54H), 2.01-2.08 ( m, 12H), 2.18 (s, 3H), 2.24-2.33 (m, 6H), 2.74-2.80 (m, 6H), 3.18-3.35 (m, 4H), 5.29-5.42 (m, 12H).
Process 6
Example 8 In the same manner as in Step 2, (9Z, 9'Z, 12Z, 12'Z) -2- (dimethylamino) -2-(((9Z, 12Z) -octadec-9,12-dienoyloxy) (9Z, 12Z) -N- (3- (methyl ((9Z, 12Z) -octadeca-9,12) obtained in step 5 instead of methyl) propane-1,3-diyl dioctadec-9,12-dienoate -Dienyl) amino) propyl) -N-((9Z, 12Z) -octadeca-9,12-dienyl) octadeca-9,12-dienamide (0.100 g, 0.118 mmol) to give the title compound (0.0804 g, 0.0895 mol, yield 76%).
ESI-MS m / z: 862 (M) ⁺ ; ¹ H-NMR (CDCl ₃ ) δ: ¹ H-NMR (CDCl ₃ ) δ: 0.88 (t, J = 7.0 Hz, 9H), 1.22-1.41 (m , 46H), 1.49-1.78 (m, 6H), 1.93-2.10 (m, 14H), 2.30 (t, J = 7.6 Hz, 2H), 2.74-2.79 (m, 6H), 3.24-3.35 (m, 8H ), 3.36-3.47 (m, 4H), 3.59-3.67 (m, 2H), 5.28-5.42 (m, 12H).

(R) -2-((2R, 3R, 4S) -3,4-bis ((9Z, 12Z) -octadeca-9,12-dienyloxy) tetrahydrofuran-2-yl) -N, N, N-trimethyl- 2-((9Z, 12Z) -octadeca-9,12-dienyloxy) ethaneaminium chloride (compound V'-1)
Process 1
Pyridine of (2R, 3R, 4S) -2-((R) -1,2-dihydroxyethyl) tetrahydrofuran-3,4-diol (Sigma-Aldrich, 0.315 g, 1.92 mmol) (Wako Pure Chemical Industries, Ltd.) 4,4'-dimethoxytrityl chloride (0.704 g, 2.02 mmol) and N, N-dimethylaminopyridine (0.047 g, 0.384 mmol) were added to the solution, and the mixture was stirred at 50 ° C overnight. After cooling to room temperature, the mixture was concentrated under reduced pressure. The resulting residue was purified by amino silica gel column chromatography (chloroform / methanol = 90/10) to obtain (2R, 3R, 4S) -2-((R) -2- (bis (4-methoxyphenyl) phenyl) Methoxy) -1-hydroxyethyl) tetrahydrofuran-3,4-diol (0.465 g, 0.997 mmol, yield 52%) was obtained.
¹ H-NMR (CDCl ₃ ) δ: 1.67-1.74 (m, 1H), 2.73-2.77 (m, 1H), 3.31 (dd, J = 9.8, 6.2 Hz, 1H), 3.41-3.50 (m, 2H) , 3.70 (dd, J = 9.6, 1.3 Hz, 1H), 3.79 (s, 6H), 3.94 (dd, J = 6.2, 3.5 Hz, 1H), 4.10-4.24 (m, 3H), 4.26-4.30 (m , 1H), 6.81-6.86 (m, 4H), 7.20-7.36 (m, 7H), 7.41-7.45 (m, 2H).
Process 2
(2R, 3R, 4S) -2-((R) -2- (bis (4-methoxyphenyl) phenyl) methoxy) -1-hydroxyethyl) tetrahydrofuran-3,4-diol (0.0669) obtained in Step 1. g, 0.143 mmol) in tetrahydrofuran (1 mL), (9Z, 12Z) -octadeca-9,12-dienyl methanesulfonate (0.247 g, 0.717 mmol), sodium hydride (oil, 60%, 0.0459 g, 1.15 mmol) was added, and the mixture was stirred overnight with heating under reflux. After cooling to room temperature, saturated brine was added to the reaction solution, and the mixture was extracted with hexane. The organic layer was washed with saturated brine, dried over anhydrous magnesium sulfate, filtered, and concentrated under reduced pressure. The obtained residue was purified by amino silica gel column chromatography (hexane / ethyl acetate = 90/10) to give (2R, 3R, 4S) -2-((R) -2- (bis (4-methoxyphenyl) ( (Phenyl) methoxy) -1-((9Z, 12Z) -octadeca-9,12-dienyloxy) ethyl) -3,4-bis ((9Z, 12Z) -octadeca-9,12-dienyloxy) tetrahydrofuran Got.
Obtained (2R, 3R, 4S) -2-((R) -2- (bis (4-methoxyphenyl) (phenyl) methoxy) -1-((9Z, 12Z) -octadeca-9,12-dienyloxy ) Ethyl) -3,4-bis ((9Z, 12Z) -octadeca-9,12-dienyloxy) tetrahydrofuran, add dichloromethane (1 mL), trifluoroacetic acid (0.0500 mL, 0.649 mmol), Stir at room temperature for 5 minutes. The reaction mixture was concentrated under reduced pressure, and the resulting residue was purified by silica gel chromatography (hexane / ethyl acetate = 70/30) to give (R) -2-((2R, 3R, 4S) -3,4-bis. ((9Z, 12Z) -octadeca-9,12-dienyloxy) tetrahydrofuran-2-yl) -2-((9Z, 12Z) -octadeca-9,12-dienyloxy) ethanol (0.0531 g, 0.0584 mmol, yield 41 %).
¹ H-NMR (CDCl ₃ ) δ: 0.89 (t, J = 7.0 Hz, 9H), 1.23-1.43 (m, 48H), 1.50-1.62 (m, 6H), 2.01-2.09 (m, 12H), 2.32 (dd, J = 8.2, 4.2 Hz, 1H), 2.74-2.80 (m, 6H), 3.37-3.50 (m, 4H), 3.54-3.69 (m, 3H), 3.69-3.77 (m, 2H), 3.80 -3.87 (m, 2H), 3.88-3.95 (m, 2H), 4.06 (dd, J = 9.8, 4.7 Hz, 1H), 5.28-5.42 (m, 12H).
Process 3
(R) -2-((2R, 3R, 4S) -3,4-bis ((9Z, 12Z) -octadeca-9,12-dienyloxy) tetrahydrofuran-2-yl) -2-obtained in step 2 To a solution of ((9Z, 12Z) -octadeca-9,12-dienyloxy) ethanol (0.0491 g, 0.0540 mmol) in dichloromethane (1 mL), methanesulfonyl chloride (manufactured by Junsei Co., Ltd., 0.0500 mL, 0.642 mmol), triethylamine ( 0.150 mL, 1.08 mmol) was added, and the mixture was stirred at room temperature for 1 hour. Methanesulfonyl chloride (0.0500 mL, 0.642 mmol) and triethylamine (0.150 mL, 1.08 mmol) were added to the reaction solution, and the mixture was stirred at room temperature for 1 hour. Chloroform (1 mL) was added to the reaction solution, and the mixture was stirred at room temperature for 1 hour. Methanesulfonyl chloride (0.0500 mL, 0.642 mmol) and triethylamine (0.150 mL, 1.08 mmol) were added to the reaction solution, and the mixture was stirred at 40 ° C. for 2 hours, and then stirred for 2 hours with heating under reflux. Saturated saline was added to the reaction solution, and the mixture was extracted with chloroform. The organic layer was dried over anhydrous magnesium sulfate, filtered and concentrated under reduced pressure. Tetrahydrofuran (1 mL) and dimethylamine (2.0 mol / L tetrahydrofuran solution, 2 mL, 2.00 mmol) were added to the obtained residue, and the mixture was stirred at 130 ° C. for 5 hours using a microwave reactor. The reaction solution was concentrated under reduced pressure. The resulting residue was purified by amino silica gel column chromatography (hexane / ethyl acetate = 95/5) to give (R) -2-((2R, 3R, 4S) -3,4-bis ((9Z, 12Z) A crude product of -octadeca-9,12-dienyloxy) tetrahydrofuran-2-yl) -N, N-dimethyl-2-((9Z, 12Z) -octadeca-9,12-dienyloxy) ethanamine was obtained. Obtained (R) -2-((2R, 3R, 4S) -3,4-bis ((9Z, 12Z) -octadeca-9,12-dienyloxy) tetrahydrofuran-2-yl) -N, N-dimethyl Chloroform (0.5 mL) and methyl iodide (1.00 mL, 16.0 mmol) were added to the crude product of -2-((9Z, 12Z) -octadeca-9,12-dienyloxy) ethanamine, and the mixture was stirred at room temperature for 2 hours. The reaction solution was concentrated under reduced pressure. Dissolve the residue in a small amount of methanol-chloroform (1: 1) and use it as an ion exchange resin (Dow Chemical, Dowex (TM) 1x-2 100 mesh, Cl type, approximately 20 times the volume, pre-washed with water and methanol). Loaded and eluted with methanol-chloroform (1: 1). The eluate was concentrated under reduced pressure, and the resulting residue was purified by silica gel column chromatography (chloroform / methanol = 90/10) to obtain the title compound (0.0130 g, 0.0132 mmol, yield 24%).
ESI-MS m / z: 951 (M) ⁺ ; ¹ H-NMR (CDCl ₃ ) δ: 0.89 (t, J = 7.0 Hz, 9H), 1.23-1.40 (m, 48H), 1.51-1.61 (m, 6H), 2.01-2.09 (m, 12H), 2.74-2.80 (m, 6H), 3.34-3.68 (m, 17H), 3.70-3.74 (m, 1H), 3.81-3.84 (m, 1H), 3.93- 4.02 (m, 3H), 4.07-4.12 (m, 1H), 5.28-5.43 (m, 12H).

Reference example 1
N, N-dimethyl-2,3-bis (((9Z, 12Z) -octadeca-9,12-dien-1-yl) oxy) propan-1-amine (compound CL-1)
CL-1 was synthesized by the method described in “Junal Controlled Release (J. Control. Release.)”, 2005, Vol. 107, p.276-287.

Reference example 2
N-methyl-N, N-bis (2-((Z) -hexadec-9-enyloxy) ethyl) amine (compound CL-2)
To a toluene (2 mL) suspension of sodium hydride (oil, 60%, 222 mg, 5.55 mmol) in a toluene (2 mL) solution of N-methyldiethanolamine (Tokyo Chemical Industry Co., Ltd., 82.6 mg, 0.693 mmol) Was added with stirring, and a solution of (Z) -hexadeca-9-enyl methanesulfonate (530 mg, 1.66 mmol) in toluene (2 mL) was added dropwise. The resulting mixture was stirred for 2 hours under heating to reflux. After cooling to room temperature, the reaction was quenched with water. To the obtained mixture was added saturated brine, and the mixture was extracted with ethyl acetate. The organic layer was dried over anhydrous magnesium sulfate and concentrated under reduced pressure. The residue was purified by silica gel column chromatography (chloroform / methanol = 100/0 to 98/2) to give the title compound (199 mg, 0.353 mmol, yield 51%).
ESI-MS m / z: 565 (M + H) ⁺ ;

Reference Example 3
trans-1-Methyl-3,4-bis ((((Z) -octadec-9-en-1-yl) oxy) methyl) pyrrolidine (Compound CL-3)
CL-3 was synthesized by the method described in International Publication No. 2011/136368.

Reference example 4
trans-1-methylpyrrolidine-3,4-diyl) bis (methylene) (9Z, 9'Z, 12Z, 12'Z) -bis (octadeca-9,12-dienoate) (compound CL-4)
CL-4 was synthesized by the method described in International Publication No. 2011/136368.

Reference Example 5
(6Z, 9Z, 28Z, 31Z) -Heptatriaconta-6,9,28,31-tetraen-19-yl 4- (dimethylamino) butanoate (Compound CL-5)
CL-5 was synthesized by a method according to the method described in International Publication No. 2010/054401.
ESI-MS m / z: 642

Reference Example 6
3- (Di ((9Z, 12Z) -octadeca-9,12-dien-1-yl) amino) propan-1-ol (Compound CL-6)
CL-6 was synthesized by the method described in International Publication No. 2014/007398.

Reference Example 7
(9Z, 12Z) -N- (2-(((Z) -Octadeca-9-en-1-yl) oxy) ethyl) octadeca-9,12-dien-1-amine (Compound CL-7)
CL-7 was synthesized by the method described in International Publication No. 2014/007398.

Reference Example 8
1-Methyl-3,3-di ((9Z, 12Z) -octadeca-9,12-dien-1-yl) azetidine (Compound CL-8)
CL-8 was synthesized by the method described in International Publication No. 2016/002753.

Reference Example 9
N, 2-Dimethyl-1,3-bis (((9Z, 12Z) -octadeca-9,12-dien-1-yl) oxy) propan-2-amine (Compound CL-9)
Process 1
2-Amino-2-methylpropane-1,3-diol (Tokyo Chemical Industry Co., Ltd., 0.300 g, 4.76 mmol) was dissolved in tetrahydrofuran (3 mL) and sodium hydride (oil 60%, 0.171 g, 7.13 mmol) ) Was added at room temperature. After the foaming stopped, (9Z, 12Z) -octadeca-9,12-dienyl methanesulfonate (manufactured by Nu-Chek Prep, Inc., 2.458 g, 7.13 mmol) was added, and the mixture was stirred for 2 hours under heating reflux. A saturated aqueous ammonium chloride solution was added to the reaction mixture, and the mixture was extracted with ethyl acetate. The organic layer was washed with saturated brine, dried over anhydrous magnesium sulfate, and filtered. The filtrate was concentrated under reduced pressure, and the resulting residue was purified by silica gel column chromatography (chloroform / methanol = 100/0 to 90/10) to give 2-methyl-1,3-bis ((9Z, 12Z) -Octadeca-9,12-dien-1-yloxy) propan-2-amine (0.280 g, yield 16%) was obtained.
ESI-MS m / z: 602 (M + H) ⁺
Process 2
2-Methyl-1,3-bis ((9Z, 12Z) -octadeca-9,12-dien-1-yloxy) propan-2-amine (0.500 g, 0.831 mmol) obtained in Step 1 was added to dichloromethane (3 mL ), Triethylamine (Wako Pure Chemical Industries, 2.55 mL, 18.3 mmol) and 2-nitrobenzene-1-sulfonyl chloride (Sigma-Aldrich, 0.368 g, 1.66 mmol) were added under ice-cooling, and the mixture was returned to room temperature. After that, the mixture was stirred for 1 hour. Water was added to the reaction mixture, and the mixture was extracted with hexane. The organic layer was washed with saturated brine, dried over anhydrous magnesium sulfate, and filtered. The filtrate was concentrated under reduced pressure, and the obtained residue was purified by silica gel column chromatography (hexane / ethyl acetate = 99/1 to 85/15) to give N- (2-methyl-1,3-bis (( 9Z, 12Z) -octadeca-9,12-dien-1-yloxy) propan-2-yl) -2-nitrobenzenesulfonamide (0.400 g, 61% yield) was obtained.
ESI-MS m / z: 787 (M + H) ⁺
Process 3
N- (2-Methyl-1,3-bis ((9Z, 12Z) -octadeca-9,12-dien-1-yloxy) propan-2-yl) -2-nitrobenzenesulfonamide obtained in Step 2 ( 0.200 g, 0.274 mmol) was dissolved in tetrahydrofuran (3 mL), and cesium carbonate (Wako Pure Chemical Industries, 0.248 g, 0.726 mmol) and methyl iodide (Tokyo Chemical Industry, 0.048 mL, 0.762 mmol) were added. In addition, the mixture was stirred at 70 ° C. for 1 hour using a microwave reactor. Water was added to the reaction mixture, and the mixture was extracted with hexane. The organic layer was washed with saturated brine, dried over anhydrous magnesium sulfate, and filtered. The filtrate was concentrated under reduced pressure, and N-methyl-N- (2-methyl-1,3-bis (((9Z, 12Z) -octadeca-9,12-dien-1-yl) oxy) propane as a crude product -2-yl) -2-nitrobenzenesulfonamide (0.200 g, 91% yield) was obtained.
ESI-MS m / z: 801 (M + H) ⁺
Process 4
N-methyl-N- (2-methyl-1,3-bis ((9Z, 12Z) -octadeca-9,12-dien-1-yloxy) propan-2-yl) -2-obtained in step 3 Nitrobenzenesulfonamide (0.200 g, 0.250 mmol) is dissolved in acetonitrile (2 mL), 1-dodecanethiol (Tokyo Chemical Industry Co., Ltd., 0.149 mL, 0.624 mmol), 1,8-diazabicyclo [5.4.0] -7 -Undecene (manufactured by Nacalai Tesque, 0.0940 mL, 0.624 mmol) was added and stirred at 80 ° C. for 1 hour. Water was added to the reaction mixture, and the aqueous layer was extracted with ethyl acetate. The organic layer was washed with saturated brine, dried over anhydrous magnesium sulfate, filtered, and concentrated under reduced pressure. The obtained residue was purified by NH silica gel column chromatography (hexane / ethyl acetate = 90/10 to 75/25) to give compound CL-9 (0.070 g, yield 46%).
ESI-MS m / z: 616 (M + H) ⁺ ; ¹ H-NMR (CDCl ₃ ) δ: 0.89 (t, J = 6.8 Hz, 6H), 1.02 (s, 3H), 1.25-1.40 (m, 32H), 1.50-1.59 (m, 4H), 2.05 (q, J = 6.8 Hz, 8H), 2.32 (s, 3H), 2.77 (t, J = 6.3 Hz, 4H), 3.26 (s, 4H), 3.40 (t, J = 6.6 Hz, 4H), 5.28-5.43 (m, 8H).

Reference Example 10
Methyldi ((9Z, 12Z) -octadeca-9,12-dienyl) amine (Compound CL-10)
(9Z, 12Z) -octadeca-9,12-dienyl methanesulfonate (1.03 g, 3.00 mmol) was added to methylamine (Sigma-Aldrich, approximately 2 mol / L tetrahydrofuran solution, 10.5 mL, 21.0 mmol), The mixture was heated and stirred at 150 ° C. for 90 minutes using a microwave reactor. The reaction solution is diluted with ethyl acetate, washed with 2 mol / L aqueous sodium hydroxide solution and then with saturated brine, dried over anhydrous magnesium sulfate, filtered, and concentrated under reduced pressure to give methyl ((9Z, 12Z) -octadeca A crude product of -9,12-dienyl) amine was obtained.
To the obtained crude product, add (9Z, 12Z) -octadeca-9,12-dienyl methanesulfonate (0.93 g, 2.70 mmol) and 50% aqueous sodium hydroxide solution (0.960 g, 12.0 mmol). The mixture was heated and stirred at 135 ° C. for 60 minutes. After cooling to room temperature, the reaction mixture was diluted with ethyl acetate, washed with water and then with saturated brine, dried over anhydrous magnesium sulfate, filtered, and concentrated under reduced pressure. The obtained residue was purified by silica gel column chromatography (chloroform / methanol = 100/0 to 97/3) to obtain Compound CL-10 (1.07 g, 2.03 mmol, overall yield 67%).
ESI-MS m / z: 529 (M + H) ⁺ ; ¹ H-NMR (CDCl ₃ ) δ: 0.89 (t, J = 6.7 Hz, 6H), 1.29 (br s, 32H), 1.40-1.51 (m , 4H), 1.97-2.06 (m, 8H), 2.20 (s, 3H), 2.30 (t, J = 7.6 Hz, 4H), 2.77 (t, J = 5.8 Hz, 4H), 5.28-5.43 (m, 8H).

Reference Example 11
N-methyl-2-(((9Z, 12Z) -octadeca-9,12-dien-1-yl) oxy) -N- (2-((((9Z, 12Z) -octadeca-9,12-diene- 1-yl) oxy) ethyl) ethan-1-amine (compound CL-11)
CL-11 was synthesized by the method described in International Publication No. 2011/136368.

Reference Example 12
(3R, 4R) -3,4-bis (((Z) -hexadec-9-en-1-yl) oxy) -1-methylpyrrolidine (Compound CL-12)
CL-12 was synthesized by the method described in International Publication No. 2011/136368.

Reference Example 13
2- (Dimethylamino) -N-((6Z, 9Z, 28Z, 31Z) -Heptatriconta-6,9,28,31-tetraen-19-yl) acetamide (Compound CL-13)
CL-13 was synthesized by the method described in International Publication No. 2013/059496.

Reference Example 14
3- (Dimethylamino) propane-1,2-diyl (9Z, 9'Z, 12Z, 12'Z) -bis (octadeca-9,12-dienoate) (Compound CL-14)
CL-14 was synthesized by the method described in “BioChemistry”, 1994, Vol. 33, p.12573-12580.

Reference Example 15
(9Z, 12Z) -di ((9Z, 12Z) -octadeca-9,12-dien-1-yl) amine (Compound CL-15)
CL-15 was synthesized by the method described in International Publication No. 2014/007398.

Reference Example 16
Bis (2-(((9Z, 12Z) -octadeca-9,12-dien-1-yl) oxy) ethyl) amine (Compound CL-16)
CL-16 was synthesized by the method described in International Publication No. 2011/136368.

Reference Example 17
(9Z, 12Z) -N-Methyl-N- (2-(((9Z, 12Z) -octadeca-9,12-dien-1-yl) oxy) ethyl) octadeca-9,12-dien-1-amine (Compound CL-17)
2- (Methylamino) ethanol (Tokyo Chemical Industry Co., Ltd., 0.125 g, 1.66 mmol) was dissolved in toluene (2.5 mL) and sodium hydride (oil, 60%, 0.333 g, 8.32 mmol), (9Z, 12Z ) -Octadeca-9,12-dien-1-yl methanesulfonate (manufactured by Nu-Chek Prep, Inc., 1.32 g, 3.83 mmol) in toluene (2.5 mL) was added in order, and the mixture was stirred for 2 hours under reflux with heating. The reaction mixture was cooled to room temperature, ethanol and water were added, and the aqueous layer was extracted with ethyl acetate. The organic layer was dried over anhydrous magnesium sulfate and filtered. The filtrate was concentrated under reduced pressure, and the resulting residue was purified by silica gel column chromatography (chloroform / methanol = 100/0 to 97/3) to give compound CL-17 (0.211 g, yield 22%). It was.
ESI-MS m / z: 572 (M + H) ⁺ ; ¹ H-NMR (CDCl ₃ ) δ: 0.89 (t, J = 6.9 Hz, 6H), 1.24-1.38 (m, 32H), 1.43-1.49 ( m, 2H), 1.53-1.59 (m, 2H), 2.05 (q, J = 7.2 Hz, 8H), 2.27 (s, 3H), 2.37 (t, J = 7.7 Hz, 2H), 2.57 (t, J = 6.2 Hz, 2H), 2.78 (t, J = 6.8 Hz, 4H), 3.42 (t, J = 6.8 Hz, 2H), 3.52 (t, J = 6.2 Hz, 2H), 5.30-5.41 (m, 8H ).

Reference Example 18
(9Z, 12Z) -N- (3-(((9Z, 12Z) -octadeca-9,12-dien-1-yl) oxy) propyl) octadeca-9,12-dien-1-amine (Compound CL- 18)
CL-18 was synthesized by a method according to the method described in International Publication No. 2014/007398.
ESI-MS m / z: 572

Reference Example 19
(1-Methylpiperidin-3-yl) methyl di ((11Z, 14Z) -icosa-11,14-dien-1-yl) carbamate (Compound CL-19)
CL-19 was synthesized by the method described in International Publication No. 2014/007398.

Reference Example 20
(13Z, 16Z) -N, N-Dimethyl-4-((9Z, 12Z) -octadeca-9,12-dien-1-yl) docosa-3,13,16-trien-1-amine (Compound CL- 20)
Process 1
To a solution of heptatriaconta-6,9,28,31-tetraen-19-one (0.353 g, 0.186 mmol) synthesized in the method described in WO2009 / 132131 in tetrahydrofuran (0.882 mL) under an argon atmosphere, Anhydrous cerium (III) chloride (manufactured by Tokyo Chemical Industry Co., Ltd., 0.174 g, 0.706 mmol) was added. Thereafter, cyclopropylmagnesium bromide (manufactured by Sigma-Aldrich, 0.5 mmol / L. 1.06 mL, 0.529 mmol) was added under ice cooling, and the mixture was stirred for 5 minutes and then stirred at room temperature for 1 hour. A saturated aqueous ammonium chloride solution was added to the reaction mixture, and the aqueous layer was extracted with ethyl acetate. The organic layer was dried over anhydrous magnesium sulfate and filtered. The filtrate was concentrated under reduced pressure, and the resulting residue was purified by silica gel column chromatography (hexane / ethyl acetate = 97 / 3-94 / 6) to give (6Z, 9Z, 28Z, 31Z) -19-cyclopropyl. Heptatriaconta-6,9,28,31-tetraen-19-ol (0.141 g, yield 70%) was obtained.
ESI-MS m / z: 569
Process 2
Dichloromethane (2 mL) of (6Z, 9Z, 28Z, 31Z) -19-cyclopropylheptatrita-6,9,28,31-tetraen-19-ol (0.141 g, 0.248 mmol) obtained in step 1 Lithium bromide (Sigma-Aldrich, 0.108 g, 1.24 mmol) and chlorotrimethylsilane (Tokyo Kasei Kogyo, 0.135 g, 1.24 mmol) were added to the solution at room temperature, and the mixture was stirred for 1 hour. Thereafter, lithium bromide (Sigma-Aldrich, 0.108 g, 1.24 mmol) and chlorotrimethylsilane (Tokyo Kasei Kogyo, 0.135 g, 1.24 mmol) were additionally added, and the mixture was stirred for 1 hour.
A saturated aqueous sodium hydrogen carbonate solution was added to the reaction mixture, and the aqueous layer was extracted with hexane. The organic layer was dried over anhydrous magnesium sulfate and filtered. The filtrate was concentrated under reduced pressure, and the obtained residue was purified by silica gel column chromatography (hexane / ethyl acetate = 100/0 to 90/10 to give (6Z, 9Z, 28Z, 31Z) -19- (3- Bromopropylidene) heptatriaconta-6,9,28,31-tetraene (0.074 g, 47% yield) was obtained.
ESI-MS m / z: 632
Process 3
The (6Z, 9Z, 28Z, 31Z) -19- (3-bromopropylidene) heptatriconta-6,9,28,31-tetraene (0.074 g, 0.117 mmol) obtained in Step 2 was added to dimethylamine (Sigma -Aldrich, 2.0 mmol / L tetrahydrofuran solution, 1.5 mL, 3.0 mmol) was added, and the mixture was heated and stirred at 130 ° C. for 90 minutes under microwave irradiation. A saturated aqueous sodium hydrogen carbonate solution was added to the reaction mixture, and the aqueous layer was extracted with hexane. The organic layer was washed with saturated brine, dried over anhydrous magnesium sulfate and filtered. The filtrate was concentrated under reduced pressure, and the obtained residue was purified by NH silica gel column chromatography (hexane / ethyl acetate = 97 / 3-88 / 12) to obtain CL-20 (0.062 g, yield 69%). It was.
ESI-MS m / z: 596

Reference Example 21
(S) -2-Amino-3-hydroxy-N, N-bis (2-(((Z) -octadeca-9-en-1-yl) oxy) ethyl) propanamide (Compound CL-21)
CL-21 was synthesized by the method described in International Publication No. 2011/136368.

Reference Example 22
(3R, 4R) -3,4-bis (((11Z, 14Z) -icosa-11,14-dien-1-yl) oxy) pyrrolidine (Compound CL-22)
CL-22 was synthesized by the method described in International Publication No. 2011/136368.

Reference Example 23
trans-3,4-bis ((((11Z, 14Z) -icosa-11,14-dien-1-yl) oxy) methyl) -1-methylpyrrolidine (Compound CL-23)
CL-23 was synthesized by the method described in International Publication No. 2011/136368.

Reference Example 24
1-((S) -2,3-bis (((9Z, 12Z) -octadeca-9,12-dien-1-yl) oxy) propyl) pyrrolidine (Compound CL-24)
CL-24 was synthesized by the method described in International Publication No. 2009/129395.

Reference Example 25
2- (2,2-di ((9Z, 12Z) -octaeca-9,12-dien-1-yl) -1,3-dioxolan-4-yl) -N, N-dimethylethan-1-amine ( Compound CL-25)
CL-25 was synthesized by the method described in International Publication No. 2010/042877.

Reference Example 26
3- (Dimethylamino) propyl di ((9Z, 12Z) -oxadec-9,12-dien-1-yl) carbamate (Compound CL-26)
CL-26 was synthesized by the method described in International Publication No. 2014/007398.

Reference Example 27
4- (Dimethylamino) butyl di ((9Z, 12Z) -oxadec-9,12-dien-1-yl) carbamate (Compound CL-27)
CL-27 was synthesized by the method described in International Publication No. 2014/007398.

Reference Example 28
2- (Di ((9Z, 12Z) -octadeca-9,12-dien-1-yl) amino) ethane-1-ol (Compound CL-28)
CL-28 was synthesized by the method described in International Publication No. 2014/007398.

Reference Example 29
2- (Dimethylamino) -3-(((9Z, 12Z) -octadeca-9,12-dien-1-yl) oxy) -2-(((((9Z, 12Z) -octadeca-9,12-diene -1-yl) oxy) methyl) propan-1-ol (compound CL-29)
CL-29 was synthesized by the method described in International Publication No. 2011/149733.

Reference Example 30
(6Z, 9Z, 28Z, 31Z) -N, N-Dimethylheptatriaconta-6,9,28,31-tetraene-19-amine (Compound CL-30)
CL-30 was synthesized by the method described in International Publication No. 2010/054405.

Reference Example 31
N, N, 2-trimethyl-1,3-bis (((9Z, 12Z) -octadeca-9,12-dien-1-yl) oxy) propan-2-amine (Compound CL-31)
2-Methyl-1,3-bis ((9Z, 12Z) -octadeca-9,12-dien-1-yloxy) propan-2-amine (0.240 g, 0.399 mmol) obtained in Step 1 of Reference Example 9 Is dissolved in a mixed solvent of 1,2-dichloroethane (1 mL) and methanol (1 mL), and formaldehyde (Wako Pure Chemical Industries, 37% aqueous solution, 0.144 mL, 1.99 mmol), sodium triacetoxyborohydride (Tokyo) Kasei Kogyo Co., Ltd., 0.211 g, 0.997 mmol) was added, and the mixture was stirred overnight at room temperature. Water was added to the reaction mixture, and the aqueous layer was extracted with ethyl acetate. The organic layer was washed with saturated sodium bicarbonate, dried over anhydrous magnesium sulfate and filtered. The filtrate was concentrated under reduced pressure, and the obtained residue was purified by NH silica gel column chromatography (hexane / ethyl acetate = 99 / 1-80 / 20) to give compound 2 (0.191 g, yield 76%). It was.
ESI-MS m / z: 630 (M + H) ⁺ ; ¹ H-NMR (CDCl ₃ ) δ: 0.89 (t, J = 7.0 Hz, 6H), 0.95 (s, 3H), 1.26-1.39 (m, 32H), 1.53-1.58 (m, 4H), 2.05 (q, J = 6.9 Hz, 8H), 2.31 (s, 6H), 2.77 (t, J = 6.3 Hz, 4H), 3.33-3.42 (m, 8H ), 5.27-5.43 (m, 8H).

Reference Example 32
N-methyl-2-(((Z) -octadeca-6-en-1-yl) oxy) -N- (2-(((Z) -octadeca-6-en-1-yl) oxy) ethyl) Ethan-1-amine (Compound CL-32)
CL-32 was synthesized by the method described in International Publication No. 2011/136368.

Reference Example 33
(3R, 4R) -3- (Dimethylamino) propyl 3,4-bis ((9Z, 12Z) -octadeca-9,12-dienyloxy) pyrrolidine-1-carboxylate (Compound CL-33)
Process 1
To a suspension of sodium hydride (oil, 60%, 5.80 g, 145 mmol) in toluene (100 mL), (3R, 4R) -1-benzylpyrrolidine-3,4-diol (Diverchim SA, 3.50 g , 18.1 mmol) in toluene (70 mL) was added with stirring, and (9Z, 12Z) -octadeca-9,12-dienyl methanesulfonate (15.6 g, 45.3 mmol) in toluene (30 mL) was added. It was dripped. The resulting mixture was stirred overnight with heating under reflux. After cooling to room temperature, the reaction was quenched with a saturated aqueous ammonium chloride group solution. Saturated brine was added to the resulting mixture, and the mixture was extracted twice with ethyl acetate. The organic layers were combined, dried over anhydrous magnesium sulfate, and concentrated under reduced pressure. The residue was purified by silica gel column chromatography (methanol / chloroform = 0/100 to 2/98) to give (3R, 4R) -1-benzyl-3,4-bis ((9Z, 12Z) -octadeca-9, 12-Dienyloxy) pyrrolidine (6.96 g, 10.1 mmol, yield 56%) was obtained.
ESI-MS m / z: 691 (M + H) ⁺ ; ¹ H-NMR (CDCl ₃ ) δ: 0.89 (t, J = 6.9 Hz, 6H), 1.26-1.38 (m, 30H), 1.52-1.62 ( m, 6H), 2.05 (q, J = 6.3 Hz, 8H), 2.50 (dd, J = 9.9, 4.3 Hz, 2H), 2.77 (t, J = 5.8 Hz, 4H), 2.85 (dd, J = 9.6 , 5.9 Hz, 2H), 3.37-3.45 (m, 4H), 3.52-3.66 (m, 2H), 3.83 (t, J = 4.6 Hz, 2H), 5.28-5.43 (m, 8H), 7.23-7.33 ( m, 5H).
Process 2
The (3R, 4R) -1-benzyl-3,4-bis ((9Z, 12Z) -octadeca-9,12-dienyloxy) pyrrolidine (6.96 g, 10.1 mmol)) obtained in Step 1 was replaced with 1,2- It was dissolved in dichloroethane (100 mL), 1-chloroethyl chloroformate (3.30 ml, 30.3 mmol) was added, and the mixture was stirred at 130 ° C. for 1 hr. Methanol (100 mL) was added to the reaction solution, and the mixture was further stirred at 130 ° C. for 1 hour. After cooling to room temperature, the mixture was concentrated under reduced pressure, and the resulting residue was purified by silica gel column chromatography (chloroform / methanol = 100/0 to 92/8). The obtained organic layer was washed with a saturated aqueous sodium hydrogen carbonate solution and then with a saturated aqueous sodium chloride solution, dried over anhydrous magnesium sulfate, filtered, and concentrated under reduced pressure to give (3R, 4R) -3,4-bis ((9Z , 12Z) -octadeca-9,12-dienyloxy) pyrrolidine (5.56 g, 9.27 mmol, 92% yield).
ESI-MS m / z: 601 (M + H) ⁺ ; ¹ H-NMR (CDCl ₃ ) δ: 0.89 (t, J = 6.9 Hz, 6H), 1.29-1.41 (m, 30H), 1.49-1.60 ( m, 4H), 1.67 (br s, 3H), 2.05 (q, J = 6.5 Hz, 8H), 2.75-2.85 (m, 6H), 3.09 (dd, J = 12.4, 5.1 Hz, 2H), 3.37- 3.49 (m, 4H), 3.76 (dd, J = 5.0, 3.3 Hz, 2H), 5.28-5.43 (m, 8H).
Process 3
In the same manner as in Step 2 of Reference Example 4, instead of di ((Z) -octadeca-9-enyl) amine, (3R, 4R) -3,4-bis ((9Z, 12Z) obtained in Step 2 was used. ) -Octadeca-9,12-dienyloxy) pyrrolidine (0.111 g, 0.185 mmol) was used to obtain the title compound (0.101 g, 0.139 mmol, 75%).
ESI-MS m / z: 730 (M + H) ⁺ ; ¹ H-NMR (CDCl ₃ ) δ: 0.89 (t, J = 6.9 Hz, 6H), 1.24-1.40 (m, 32H), 1.50-1.57 ( m, 4H), 1.77-1.83 (m, 2H), 2.02-2.08 (m, 8H), 2.23 (s, 6H), 2.34 (t, J = 7.4 Hz, 2H), 2.77 (t, J = 6.8 Hz , 4H), 3.38-3.56 (m, 8H), 3.83-3.86 (m, 2H), 4.11 (t, J = 6.5 Hz, 2H), 5.30-5.42 (m, 8H).

Reference Example 34
(9Z, 12Z) -N- (2-(((9Z, 12Z) -octadeca-9,12-dien-1-yl) oxy) ethyl) octadeca-9,12-dien-1-amine (compound CL- 34)
CL-34 was synthesized by the method described in International Publication No. 2014/007398.

Reference Example 35
(9Z, 12Z) -N- (2-((((Z) -Hexadeca-9-en-1-yl) oxy) ethyl) octadeca-9,12-dien-1-amine (Compound CL-35)
CL-35 was synthesized by the method described in International Publication No. 2014/007398.

Reference Example 36
N, N-dimethyl-1,3-bis (((9Z, 12Z) -octadeca-9,12-dien-1-yl) oxy) propan-2-amine (Compound CL-36)
CL-36 was synthesized by the method described in International Publication No. 2009/129385.

Reference Example 37
3- (Dimethylmino) propyl di ((Z) -octadeca-9-enyl) carbamate (Compound CL-37)
Process 1
Add (Z) -octadeca-9-enyl methanesulfonate (1.04 g, 3.00 mmol) to ammonia (Tokyo Chemical Industry Co., Ltd., approx. 2 mol / L methanol solution, 12.0 mL, 24.0 mmol) And stirred at 130 ° C. for 3 hours. A saturated aqueous sodium hydrogen carbonate solution was added to the reaction mixture, and the mixture was extracted 5 times with chloroform. The organic layers were combined, washed with saturated brine, dried over anhydrous magnesium sulfate, filtered, and concentrated under reduced pressure to obtain a crude product of (Z) -octadeca-9-enylamine.
(Z) -octadeca-9-enyl methanesulfonate (0.832 g, 2.40 mmol) and 50% aqueous sodium hydroxide solution (0.960 g, 12.0 mmol) were added to the resulting crude product, and the mixture was heated at 110 ° C. on an oil bath at 60 ° C. Stir for minutes. After cooling to room temperature, the reaction mixture was diluted with ethyl acetate, washed with water and then with saturated brine, dried over anhydrous magnesium sulfate, filtered, and concentrated under reduced pressure. The obtained residue was purified by silica gel column chromatography (chloroform / methanol = 100/0 to 95/5) to give di ((Z) -octadeca-9-enyl) amine (0.562 g, 1.085 mmol, yield) 36%) was obtained.
ESI-MS m / z: 519 (M + H) ⁺ ; ¹ H-NMR (CDCl ₃ ) δ: 0.88 (t, J = 6.7 Hz, 6H), 1.29 (br s, 45H), 1.41-1.52 (m , 4H), 1.97-2.05 (m, 8H), 2.58 (t, J = 7.2 Hz, 4H), 5.28-5.40 (m, 4H).
Process 2
Di ((Z) -octadeca-9-enyl) amine (0.156 g, 0.301 mmol) obtained in Step 1 was dissolved in chloroform (3 mL), and “Journal of American Chemical Society (J. Am. Chem. Soc.) ”, 1981, Vol. 103, p. 4194-4199, 3- (dimethylamino) propyl 4-nitrophenyl carbonate hydrochloride (0.138 g, 0.452 mmol) And triethylamine (0.168 mL, 1.21 mmol) were added, and the mixture was stirred at 110 ° C. for 60 minutes using a microwave reactor. 3- (Dimethylamino) propyl 4-nitrophenyl carbonate hydrochloride (22.9 mg, 0.0753 mmol) was added to the reaction solution, and the mixture was stirred at 110 ° C. for 20 minutes using a microwave reactor. 3- (Dimethylamino) propyl 4-nitrophenyl carbonate hydrochloride (22.9 mg, 0.0753 mmol) was added to the reaction solution, and the mixture was stirred at 110 ° C. for 20 minutes using a microwave reactor. 3- (Dimethylamino) propyl 4-nitrophenyl carbonate hydrochloride (22.9 mg, 0.0753 mmol) was added to the reaction solution, and the mixture was stirred at 110 ° C. for 20 minutes using a microwave reactor. The reaction mixture was diluted with chloroform, washed with saturated aqueous sodium hydrogen carbonate solution, then washed with saturated brine, dried over anhydrous magnesium sulfate, filtered, and concentrated under reduced pressure. The obtained residue was dissolved in a small amount of n-hexane / ethyl acetate (1/4), adsorbed on a pad of amino-modified silica gel, eluted with n-hexane / ethyl acetate (1/4), and concentrated under reduced pressure. The obtained residue was purified by silica gel column chromatography (chloroform / methanol = 100/0 to 95/5) to give the title compound (0.173 g, 0.267 mmol, yield 89%).
ESI-MS m / z: 648 (M + H) ⁺ ; ¹ H-NMR (CDCl ₃ ) δ: 0.88 (t, J = 6.6 Hz, 6H), 1.28 (br s, 44H), 1.45-1.55 (m , 4H), 1.75-1.85 (m, 2H), 1.97-2.04 (m, 8H), 2.23 (s, 6H), 2.34 (t, J = 7.6 Hz, 2H), 3.13-3.24 (m, 4H), 4.10 (t, J = 6.4 Hz, 2H), 5.28-5.40 (m, 4H).

Reference Example 38
(11Z, 14Z) -N- (2-(((Z) -Octadeca-9-en-1-yl) oxy) ethyl) icosa-11,14-dien-1-amine (Compound CL-38)
CL-38 was synthesized by a method according to the method described in International Publication No. 2014/007398.
ESI-MS m / z: 588

Reference Example 39
(9Z, 12Z) -N- (2-(((Z) -Icosa-11-en-1-yl) oxy) ethyl) octadeca-9,12-dien-1-amine (Compound CL-39)
CL-39 was synthesized by a method according to the method described in International Publication No. 2014/007398.
ESI-MS m / z: 588

Reference Example 40
(11Z, 14Z) -N- (2-(((Z) -Icosa-11-en-1-yl) oxy) ethyl icosa-11,14-dien-1-amine (Compound CL-40)
CL-40 was synthesized by a method according to the method described in International Publication No. 2014/007398.
ESI-MS m / z: 616

Reference Example 41
(Z) -N- (2-(((Z) -octadeca-9-en-1-yl) oxy) ethyl) octadeca-9-en-1-amine (Compound CL-41)
CL-41 was synthesized by a method according to the method described in International Publication No. 2014/007398.
ESI-MS m / z: 562

Reference Example 42
Bis (2-(((11Z, 14Z) -icosa-11,14-dien-1-yl) oxy) ethyl) amine (Compound CL-42)
CL-42 was synthesized by a method according to the method described in International Publication No. 2011/136368.
ESI-MS m / z: 658

Reference Example 43
(Z) -N- (2-(((Z) -octadeca-9-en-1-yl) oxy) ethyl) hexadeca-9-en-1-amine (Compound CL-43)
CL-43 was synthesized by a method according to the method described in International Publication No. 2014/007398.
ESI-MS m / z: 534

Reference Example 44
(Z) -N- (2- (Octadec-9-en-1-yloxy) ethyl) octadecan-1-amine (Compound CL-44)
CL-44 was synthesized by a method according to the method described in International Publication No. 2014/007398.
ESI-MS m / z: 564

Reference Example 45
(Z) -N- (2- (Octadec-9-en-1-yloxy) ethyl) tetradecan-1-amine (Compound CL-45)
CL-45 was synthesized by a method according to the method described in International Publication No. 2014/007398.
ESI-MS m / z: 508

Reference Example 46
3-((3R, 4R) -3,4-bis (((9Z, 12Z) -octadeca-9,12-dien-1-yl) oxy) pyrrolidin-1-yl) propane-1,2-diol ( Compound CL-46)
CL-46 was synthesized by the method described in International Publication No. 2011/136368.

Reference Example 47
Bis (2-((((Z) -octadeca-9-en-1-yl) oxy) ethyl) amine (Compound CL-47)
CL-47 was synthesized by the method described in International Publication No. 2011/136368.

Reference Example 48
3- (Bis (2-(((Z) -octadeca-9-en-1-yl) oxy) ethyl) amino) propane-1,2-diol (Compound CL-48)
CL-48 was synthesized by the method described in International Publication No. 2011/136368.

Reference Example 49
3- (Bis (2-(((Z) -octadeca-9-en-1-yl) oxy) ethyl) amino) propanamide (Compound CL-49)
CL-49 was synthesized by the method described in International Publication No. 2011/136368.

Reference Example 50
(9Z, 12Z) -N- (2- (2-(((Z) -Octadeca-9-en-1-yl) oxy) ethoxy) ethyl) octadeca-9,12-dien-1-amine (Compound CL -50)
CL-50 was synthesized by a method according to the method described in International Publication No. 2014/007398.
ESI-MS m / z: 604

Reference Example 51
Di ((Z) -non-2-en-1-yl) 9-((4- (dimethylamino) butanoyl) oxy) heptadecandioate (Compound CL-51)
CL-51 was synthesized by the method described in International Publication No. 2011/153493.

Reference Example 52
Di ((Z) -non-2-en-1-yl) 8,8 '-((((2- (dimethylamino) ethyl) thio) carbonyl) azanediyl) dioctanoate (Compound CL-52)
CL-52 was synthesized by the method described in International Publication No. 2017/023817.

Reference Example 53
2- (Dimethylamino) -N- (2-(((Z) -octadeca-9-en-1-yl) oxy) ethyl) -N-((9Z, 12Z) -octadeca-9,12-diene- 1-yl) acetamide (Compound CL-53)
CL-53 was synthesized by a method according to the method described in International Publication No. 2014/007398.

Reference Example 54
3-((2-(((Z) -octadeca-9-en-1-yl) oxy) ethyl) ((9Z, 12Z) -octadeca-9,12-dien-1-yl) amino) propane-1 -All (compound CL-54)
CL-54 was synthesized by a method according to the method described in International Publication No. 2014/007398.

Reference Example 55
1-Methyl-3,3-bis ((((9Z, 12Z) -octadeca-9,12-dien-1-yl) oxy) methyl) azetidine (Compound CL-55)
CL-55 was synthesized by the method described in International Publication No. 2012/108397.

Reference Example 56
1-Methyl-3,3-bis (2-(((9Z, 12Z) -octadeca-9,12-dien-1-yl) oxy) ethyl) azetidine (Compound CL-56)
CL-56 was synthesized by the method described in International Publication No. 2016/002753.

Reference Example 57
1-Methyl-3,3-bis (2-(((9Z, 12Z) -octadeca-9,12-dien-1-yl) oxy) propyl) azetidine (Compound CL-57)
CL-57 was synthesized by the method described in International Publication No. 2016/002753.

Reference Example 58
2- (3,3-di ((9Z, 12Z) -octadeca-9,12-dien-1-yl) azetidin-1-yl) ethan-1-ol (Compound CL-58)
CL-58 was synthesized by the method described in International Publication No. 2016/002753.

Reference Example 59
2- (3,3-di ((9Z, 12Z) -octadeca-9,12-dien-1-yl) azetidin-1-yl) propan-1-ol (compound CL-59)
CL-59 was synthesized by the method described in International Publication No. 2016/002753.

Reference Example 60
3- (Dimethylamino) propyl 3,3-di ((9Z, 12Z) -octadeca-9,12-dien-1-yl) azetidine-1-carboxylate (Compound CL-60)
CL-60 was synthesized by the method described in International Publication No. 2016/002753.

Reference Example 61
2- (Di ((Z) -octadeca-9-en-1-yl) amino) ethane-1-ol (Compound CL-61)
CL-61 was synthesized by the method described in International Publication No. 2014/007398.

Reference Example 62
3- (Di ((Z) -octadeca-9-en-1-yl) amino) propan-1-ol (Compound CL-62)
CL-62 was synthesized by the method described in International Publication No. 2014/007398.

Reference Example 63
(11Z, 14Z) -2-((Dimethylamino) methyl) -2-((9Z, 12Z) -octadeca-9,12-dien-1-yl) icosa-11,14-dien-1-ol (compound CL-63)
Process 1
Ethyl 2-cyanoacetate (0.943 mL, 8.84 mmol), hydrogenated to a solution of (9Z, 12Z) -octadeca-9,12-dien-1-ylmethanesulfonate (7.62 g, 22.1 mmol) in tetrahydrofuran (30.0 mL) Sodium (1.06 g, 26.5 mmol) and tetrabutylammonium iodide (3.27 g, 8.84 mmol) were added, and the mixture was stirred at 60 ° C. for 2 hours. Water was added to the reaction solution and extracted with heptane. The organic layer was dried over anhydrous magnesium sulfate, filtered and concentrated under reduced pressure. The obtained residue was purified by silica gel column chromatography (heptane / ethyl acetate = 99/1 to 85/15) to obtain ethyl (11Z, 14Z) -2-cyano-2-((9Z, 12Z) -octadeca-9 , 12-Dien-1-yl) icosa-11,14-dienoate (3.50 g, 5.74 mmol, yield 64.9%).
Ethyl (11Z, 14Z) -2-cyano-2-((9Z, 12Z) -octadeca-9,12-dien-1-yl) icosa-11,14-dienoate (1.50 g, 2.46 mmol) in tetrahydrofuran (10.0 mL) Lithium aluminum hydride (0.467 g, 12.3 mmol) was added to the solution, and the mixture was stirred for 30 minutes under ice cooling. Water, sodium hydroxide, and water were added to the reaction solution at a ratio of 1: 1: 3, dried over anhydrous magnesium sulfate, filtered, and concentrated under reduced pressure. The obtained residue was purified by silica gel column chromatography (chloroform / methanol = 90/10) to obtain (11Z, 14Z) -2- (aminomethyl) -2-((9Z, 12Z) -octadeca-9,12- Dien-1-yl) icosa-11,14-dien-1-ol (1.00 g, 1.75 mmol, yield 71.1%) was obtained.
ESI-MS m / z: 573 (M + H) ⁺ .
Process 2
(11Z, 14Z) -2- (Aminomethyl) -2-((9Z, 12Z) -octadeca-9,12-dien-1-yl) icosa-11,14-dien-1-ol (0.200 g, 0.350 mmol) in dichloroethane (2.00 mL) were added a paraformaldehyde solution (0.276 g, 3.50 mmol, 37% methanol solution) and sodium triacetoxyborohydride (1.48 g, 6.99 mmol), and the mixture was stirred at room temperature for 3 hours. Water was added to the reaction solution, and the mixture was extracted with chloroform. The organic layer was dried over anhydrous magnesium sulfate, filtered and concentrated under reduced pressure. The obtained residue was purified by silica gel column chromatography (chloroform / methanol = 99/1 to 85/15) to obtain the title compound (0.0280 g, 0.0470 mmol, yield 13.3%).
ESI-MS m / z: 601 (M) ⁺ ; ¹ H-NMR (CDCl ₃ ) δ: 0.89 (t, J = 6.8 Hz, 6H), 1.23-1.29 (m, 40H), 2.05 (q, J = 6.8 Hz, 8H), 2.32 (s, 6H), 2.40 (s, 2H), 2.77 (dd, J = 9.8, 3.4 Hz, 4H), 3.53 (s, 2H), 5.29-5.42 (m, 8H).

Reference Example 64
(11Z, 14Z) -2- (Dimethylamino) -2-((9Z, 12Z) -octadeca-9,12-dien-1-yl) icosa-11,14-dien-1-ol (Compound CL-64 )
Process 1
Sodium hydride (1.21 g, 30.3 mmol) in THF (30.0 mL) was added to tert-butylethyl malonate (Tokyo Chemical Industry, 2.00 mL, 10.1 mmol), tetra-n-butylammonium iodide (Nacalai Tesque) , 0.746 g, 2.02 mmol), (9Z, 12Z) -octadeca-9,12-dienyl methanesulfonate (manufactured by Nu-Chek Prep, Inc., 8.70 g, 25.2 mmol) And stirred for 2 hours under reflux. Saturated brine was added to the reaction mixture, and the mixture was extracted with ethyl acetate. The organic layer was dried over anhydrous magnesium sulfate, filtered, and the solvent was removed under reduced pressure. The obtained residue was purified by silica gel column chromatography (hexane / ethyl acetate = 95/5) to give 1-tert-butyl 3-ethyl 2,2-di ((9Z, 12Z) -octadeca-9,12- Dien-1-yl) malonic acid (5.52 g, 80.0%) was obtained.
Process 2
1-tert-butyl 3-ethyl 2,2-di ((9Z, 12Z) -octadeca-9,12-dien-1-yl) malonic acid (5.52 g, 8.06 mmol) in dichloromethane (30.0 mL) Fluoroacetic acid (5.00 mL, 64.9 mmol) was added, and the mixture was stirred at room temperature for 30 minutes. The reaction solution was concentrated under reduced pressure. The residue was partitioned between chloroform and saturated aqueous sodium bicarbonate. The organic layer was washed with saturated brine, dried over anhydrous magnesium sulfate, filtered, and the solvent was evaporated under reduced pressure. The obtained residue was purified by silica gel column chromatography (chloroform / methanol = 95/5) to give (11Z, 14Z) -2- (ethoxycarbonyl) -2-((9Z, 12Z) -octadeca-9,12- Dien-1-yl) icosa-11,14-dienoic acid (4.04 g, 6.42 mmol, 80.0%) was obtained.
Process 3
Of (11Z, 14Z) -2- (ethoxycarbonyl) -2-((9Z, 12Z) -octadeca-9,12-dien-1-yl) icosa-11,14-dienoic acid (0.284 g, 0.452 mmol) Triethylamine (0.315 mL, 2.26 mmol) and diphenylphosphoryl azide (manufactured by Tokyo Chemical Industry Co., Ltd., 0.121 mL, 0.542 mmol) were added to a toluene (3.00 mL) solution, and the mixture was stirred at room temperature for 1 hour. Water (0.0200 mL, 1.11 mmol) was added to the reaction solution, and the mixture was stirred for 5 hours with heating under reflux. Water was added to the reaction mixture, and the mixture was extracted with ethyl acetate. The organic layer was dried over anhydrous magnesium sulfate, filtered, and the solvent was removed under reduced pressure. The obtained residue was purified by silica gel column chromatography (hexane / ethyl acetate = 80/20) to obtain (11Z, 14Z) -2-amino-2-((9Z, 12Z) -octadeca-9,12-diene. Ethyl-1-yl) icosa-11,14-dienoate (0.0457 g, 0.0762 mmol, 17.0%) was obtained.
ESI-MS m / z: 601 (M + H) ⁺ .
Process 4
(11Z, 14Z) -2-Amino-2-((9Z, 12Z) -octadeca-9,12-dien-1-yl) icosa-11,14-dienoic acid ethyl ester (0.299 g, 0.498 mmol) in THF ( 3.00 mL) Lithium aluminum hydride (Pure Chemical Co., Ltd., 0.0190 g, 0.498 mmol) was added to the solution, and the mixture was stirred at room temperature for 30 minutes. Water and an aqueous sodium hydroxide solution were added to the reaction solution, insoluble matters were removed by Celite filtration, and the filtrate was concentrated under reduced pressure. The resulting residue was purified by amino silica gel column chromatography (hexane / ethyl acetate = 50/50) to give (11Z, 14Z) -2-amino-2-((9Z, 12Z) -octadeca-9,12-diene. -1-yl) icosa-11,14-dien-1-ol (0.0667 g, 0.120 mmol, 24.0%) was obtained.
ESI-MS m / z: 559 (M + H) ⁺ .
Process 5
Of (11Z, 14Z) -2-amino-2-((9Z, 12Z) -octadeca-9,12-dien-1-yl) icosa-11,14-dien-1-ol (0.0664 g, 0.119 mmol) To a dichloroethane (1.00 mL) solution, formaldehyde (0.500 mL, 6.72 mmol) and sodium triacetoxyborohydride (manufactured by Tokyo Chemical Industry Co., Ltd., 0.101 g, 0.476 mmol) were added and stirred overnight at room temperature. A saturated aqueous sodium hydrogen carbonate solution was added to the reaction solution, and the mixture was extracted with ethyl acetate. The organic layer was washed with a saturated aqueous sodium chloride solution, dried over anhydrous magnesium sulfate, filtered, and concentrated under reduced pressure. The obtained residue was purified by silica gel column chromatography (chloroform / methanol = 80/20) to obtain the title compound (0.0474 g, 0.0809 mmol, 68.0%).
ESI-MS m / z: 587 (M + H) ⁺ ; ¹ H-NMR (CDCl ₃ ) δ: 0.89 (t, J = 7.0 Hz, 6H), 1.24-1.39 (m, 40H), 2.05 (q, J = 6.8 Hz, 8H), 2.38 (s, 6H), 2.77 (t, J = 6.7 Hz, 4H), 5.29-5.42 (m, 8H).

Reference Example 65
3- (Dimethylamino) -2,2-bis ((((9Z, 12Z) -octadeca-9,12-dien-1-yl) oxy) methyl) propan-1-ol (Compound CL-65)
Dimethylamine solution (15.0 mL, 30.0 mmol, 2M in THF) was added to 2- (bromomethyl) -2- (hydroxymethyl) propane-1,3-diol (1.52 g, 7.56 mmol), and 120 μm under microwave irradiation. Stir at 15 ° C. for 15 hours. Lithium hydroxide was added to the reaction solution and filtered. The filtrate was concentrated under reduced pressure to obtain 2-((dimethylamino) methyl) -2- (hydroxymethyl) propane-1,3-diol (1.23 g, quantitative).
To a solution of 2-((dimethylamino) methyl) -2- (hydroxymethyl) propane-1,3-diol (1.23 g, 7.56 mmol) in toluene (30.0 mL), sodium hydride (0.756 g, 18.9 mmol), ( 9Z, 12Z) -Octadeca-9,12-dien-1-ylmethanesulfonate (6.51 g, 18.9 mmol) was added, and the mixture was stirred overnight with heating under reflux. Saturated saline was added to the reaction solution, and the mixture was extracted with hexane. The organic layer was dried over anhydrous magnesium sulfate, filtered and concentrated under reduced pressure. The obtained residue was purified by silica gel column chromatography (chloroform / methanol = 90/10) and amino silica gel column chromatography (hexane / ethyl acetate = 90/10) to give the title compound (1.80 g, 2.73 mmol, yield 36.1). %).
ESI-MS m / z: 661 (M) ⁺ ; ¹ H-NMR (CDCl ₃ ) δ: 0.89 (t, J = 7.0 Hz, 6H), 1.24-1.39 (m, 32H), 1.50-1.56 (m, 4H), 2.05 (q, J = 6.8 Hz, 8H), 2.30 (s, 6H), 2.53 (s, 2H), 2.77 (t, J = 6.3 Hz, 4H), 3.30-3.41 (m, 8H), 3.71 (s, 2H), 5.29-5.42 (m, 8H).

Reference Example 66
((2-((2- (Dimethylamino) ethyl) thio) acetyl) azanediyl) bis (ethane-2,1-diyl) ditetradecanoate (compound CL-66)
CL-62 was synthesized by the method described in International Publication No. 2012/170952.

II-25 obtained in Example 34, CL-1 obtained in Reference Example 1 and 1,2-dimyristoyl-sn-glycero-3-phosphoethanolamine-N- [methoxy (polyethylene glycol) -2000] Formulation 1 containing nucleic acid-containing lipid nanoparticles was produced using (PEG-DMPE), 1,2-distearoyl-sn-glycero-3-phosphocholine (DSPC), and cholesterol as follows.
The nucleic acid used is a siRNA that suppresses the expression of a luciferase (hereinafter referred to as Luc) gene consisting of a base sequence of a sense strand (5′-CCGUCGUAAUUCGUGACACAGA-3 ′) and an antisense strand (5′-UUGCUCACGAAUACGACGGUG-3 ′). Yes, obtained from GeneDesign (hereinafter "Luc siRNA"). PEG-DMPE, DSPC, and cholesterol were obtained from NOF.
II-25 was dissolved in 100% ethanol to give 5 mg / mL to obtain a lipid stock solution. CL-1, PEG-DMPE, DSPC, and cholesterol were dissolved in 100% ethanol so that each would be 20 mg / mL to obtain a lipid stock solution. Each lipid stock solution was stored at −20 ° C. and heated to 60 ° C. immediately before preparation preparation to dissolve the lipid and then returned to room temperature before use.
Luc siRNA was dissolved in water for injection at 1 mg / mL to give a Luc siRNA solution.
II-25 used as the lipid stock solution was added to 20 mL of an 80% aqueous ethanol solution to a concentration of 0.313 μmol. Subsequently, 200 μL of the Luc siRNA solution was added and stirred for 1 minute, and then CL-1 / PEG-DMPE / DSPC / Chol as the lipid stock solution was added to the solution as 1.88 μmol / 0.145 μmol / 0.515 μmol / 1.07 μmol. It added so that it might become. Thereafter, water for injection was added at a flow rate of 62 mL / second or more to form a 20% or less aqueous ethanol solution to form a crude preparation. The obtained crude preparation was concentrated using Amicon Ultra (manufactured by Millipore), further replacing the solvent with physiological saline, and filtered in a clean bench using a 0.2 μm filter (manufactured by Toyo Roshi Kaisha, Ltd.). Further, the siRNA concentration of the obtained preparation was measured, and diluted with physiological saline so that the siRNA concentration was 0.1 mg / mL, whereby preparation 1 was obtained.

Preparation 2 was obtained in the same manner as in Example 72 except that CL-1 of preparation 1 was changed to CL-2 obtained in Reference Example 2.

Preparation 3 was obtained in the same manner as in Example 72 except that CL-1 of preparation 1 was changed to CL-3 obtained in Reference Example 3.

Formulation 4 was obtained in the same manner as in Example 72 except that CL-1 of formulation 1 was changed to CL-4 obtained in Reference Example 4.

Formulation 5 was obtained in the same manner as in Example 72 except that CL-1 of formulation 1 was changed to CL-5 obtained in Reference Example 5.

Preparation 6 was obtained in the same manner as in Example 72 except that CL-1 of preparation 1 was changed to CL-6 obtained in Reference Example 6.

Preparation 7 was obtained in the same manner as in Example 72 except that CL-1 of preparation 1 was changed to CL-7 obtained in Reference Example 7.

Preparation 8 was obtained in the same manner as in Example 72 except that CL-1 of preparation 1 was changed to CL-8 obtained in Reference Example 8.

Preparation 9 was obtained in the same manner as in Example 72 except that CL-1 of preparation 1 was changed to CL-9 obtained in Reference Example 9.

Preparation 10 was obtained in the same manner as in Example 72 except that CL-1 of preparation 1 was changed to CL-10 obtained in Reference Example 10.

Preparation 11 was obtained in the same manner as in Example 72 except that CL-1 of preparation 1 was changed to CL-11 obtained in Reference Example 11.

Formulation 12 was obtained in the same manner as in Example 72 except that CL-1 of formulation 1 was changed to CL-12 obtained in Reference Example 12.

Preparation 13 was obtained in the same manner as in Example 72 except that CL-1 of preparation 1 was changed to CL-13 obtained in Reference Example 13.

Preparation 14 was obtained in the same manner as in Example 72 except that CL-1 of preparation 1 was changed to CL-14 obtained in Reference Example 14.

Preparation 15 was obtained in the same manner as in Example 72 except that CL-1 of preparation 1 was changed to CL-15 obtained in Reference Example 15.

Preparation 16 was obtained in the same manner as in Example 72 except that CL-1 of preparation 1 was changed to CL-16 obtained in Reference Example 16.

Preparation 17 was obtained in the same manner as in Example 72 except that CL-1 of preparation 1 was changed to CL-17 obtained in Reference Example 17.

Preparation 18 was obtained in the same manner as in Example 72, except that CL-1 of Preparation 1 was changed to CL-18 obtained in Reference Example 18.

Preparation 19 was obtained in the same manner as in Example 72, except that CL-1 of Preparation 1 was changed to CL-19 obtained in Reference Example 19.

Preparation 20 was obtained in the same manner as in Example 72 except that CL-1 of preparation 1 was changed to CL-20 obtained in Reference Example 20.

Preparation 21 was obtained in the same manner as in Example 72, except that CL-1 of preparation 1 was changed to CL-21 obtained in Reference Example 21.

[Comparative Example 1]
CL-1 and 1,2-dimyristoyl-sn-glycero-3-phosphoethanolamine-N- [methoxy (polyethylene glycol) -2000] (PEG-DMPE) obtained in Reference Example 1, Preparation 22 containing nucleic acid-containing lipid nanoparticles was prepared using stearoyl-sn-glycero-3-phosphocholine (DSPC) and cholesterol as follows.
The nucleic acid used is a siRNA that suppresses the expression of a luciferase (hereinafter referred to as Luc) gene consisting of a base sequence of a sense strand (5′-CCGUCGUAAUUCGUGACACAGA-3 ′) and an antisense strand (5′-UUGCUCACGAAUACGACGGUG-3 ′). Yes, obtained from GeneDesign (hereinafter "Luc siRNA"). PEG-DMPE, DSPC, and cholesterol were obtained from NOF.
CL-1, PEG-DMPE, DSPC, and cholesterol were dissolved in 100% ethanol so that each would be 20 mg / mL to obtain a lipid stock solution. Each lipid stock solution was stored at −20 ° C. and heated to 60 ° C. immediately before preparation preparation to dissolve the lipid and then returned to room temperature before use.
Luc siRNA was dissolved in water for injection at 1 mg / mL to give a Luc siRNA solution.
Add 200 μL of the above Luc siRNA solution to 20 mL of 80% ethanol aqueous solution and stir for 1 minute. It added so that it might become micromol / 1.28 micromol. Thereafter, water for injection was added at a flow rate of 62 mL / second or more to form a 20% or less aqueous ethanol solution to form a crude preparation. The obtained crude preparation was concentrated using Amicon Ultra (manufactured by Millipore), further replacing the solvent with physiological saline, and filtered in a clean bench using a 0.2 μm filter (manufactured by Toyo Roshi Kaisha, Ltd.). Further, the siRNA concentration of the obtained preparation was measured, and diluted with physiological saline so that the siRNA concentration was 0.1 mg / mL, whereby preparation 22 was obtained.

[Comparative Example 2]
A preparation 23 was obtained in the same manner as in Comparative Example 1, except that CL-1 of the preparation 22 was changed to CL-2 obtained in Reference Example 2.

[Comparative Example 3]
A preparation 24 was obtained in the same manner as in Comparative Example 1, except that CL-1 of the preparation 22 was changed to CL-3 obtained in Reference Example 3.

[Comparative Example 4]
Formulation 25 was obtained in the same manner as in Comparative Example 1, except that CL-1 in Formulation 22 was changed to CL-4 obtained in Reference Example 4.

[Comparative Example 5]
Formulation 26 was obtained in the same manner as in Comparative Example 1, except that CL-1 in Formulation 22 was changed to CL-5 obtained in Reference Example 5.

[Comparative Example 6]
Formulation 27 was obtained in the same manner as in Comparative Example 1, except that CL-1 in Formulation 22 was changed to CL-6 obtained in Reference Example 6.

[Comparative Example 7]
A preparation 28 was obtained in the same manner as in Comparative Example 1, except that CL-1 in the preparation 22 was changed to CL-7 obtained in Reference Example 7.

[Comparative Example 8]
A preparation 29 was obtained in the same manner as in Comparative Example 1, except that CL-1 of the preparation 22 was changed to CL-8 obtained in Reference Example 8.

[Comparative Example 9]
A preparation 30 was obtained in the same manner as in Comparative Example 1, except that CL-1 of the preparation 22 was changed to CL-9 obtained in Reference Example 9.

[Comparative Example 10]
Formulation 31 was obtained in the same manner as in Comparative Example 1, except that CL-1 in Formulation 22 was changed to CL-10 obtained in Reference Example 10.

[Comparative Example 11]
A preparation 32 was obtained in the same manner as in Comparative Example 1, except that CL-1 of the preparation 22 was changed to CL-11 obtained in Reference Example 11.

[Comparative Example 12]
A preparation 33 was obtained in the same manner as in Comparative Example 1, except that CL-1 of the preparation 22 was changed to CL-12 obtained in Reference Example 12.

[Comparative Example 13]
A preparation 34 was obtained in the same manner as in Comparative Example 1, except that CL-1 of the preparation 22 was changed to CL-13 obtained in Reference Example 13.

[Comparative Example 14]
A preparation 35 was obtained in the same manner as in Comparative Example 1, except that CL-1 of the preparation 22 was changed to CL-14 obtained in Reference Example 14.

[Comparative Example 15]
Formulation 36 was obtained in the same manner as in Comparative Example 1, except that CL-1 in Formulation 22 was changed to CL-15 obtained in Reference Example 15.

[Comparative Example 16]
Formulation 37 was obtained in the same manner as in Comparative Example 1, except that CL-1 in Formulation 22 was changed to CL-16 obtained in Reference Example 16.

[Comparative Example 17]
Formulation 38 was obtained in the same manner as in Comparative Example 1, except that CL-1 in Formulation 22 was changed to CL-17 obtained in Reference Example 17.

[Comparative Example 18]
Formulation 39 was obtained in the same manner as in Comparative Example 1, except that CL-1 in Formulation 22 was changed to CL-18 obtained in Reference Example 18.

[Comparative Example 19]
A preparation 40 was obtained in the same manner as in Comparative Example 1, except that CL-1 of the preparation 22 was changed to CL-19 obtained in Reference Example 19.

[Comparative Example 20]
Formulation 41 was obtained in the same manner as in Comparative Example 1, except that CL-1 in Formulation 22 was changed to CL-20 obtained in Reference Example 20.

[Comparative Example 21]
A preparation 42 was obtained in the same manner as in Comparative Example 1, except that CL-1 of the preparation 22 was changed to CL-21 obtained in Reference Example 21.

Test example 1
Measurement of average particle size of nucleic acid-containing lipid nanoparticles The average particle size of nucleic acid-containing lipid nanoparticles in the preparation was measured with a particle size measurement device (Zetasizer Nano ZS, manufactured by Malvern) (Table 27). PDI in the table represents a polydispersity index.

As a result, lipid A (II-25) and lipid B (CL-1 to 21), PEG-DMPE®, DSPC, a complex of cholesterol and nucleic acid was dispersed in ethanol, and water was rapidly added to the dispersion. Formulations 1 to 21 described in Examples 72 to 92 and Formulations 22 to 42 described in Comparative Examples 1 to 21 that do not contain lipid A were used regardless of whether lipid A (II-25) was contained or not. The average particle size was small and was 50 nm or less.

Test example 2
In vitro activity evaluation test of nucleic acid-containing lipid nanoparticles Preparations 1 to 21 described in Examples 72 to 92 and preparations 22 to 42 described in Comparative Examples 1 to 21 were forcibly expressed by the following methods, respectively. It was introduced into a cervical cancer-derived cell line HeLa cell (Luc2CP-HeLa).
Each formulation diluted with Optimem (Opti-MEM, Gibco) to a final nucleic acid concentration of 0.3 to 10 nM is dispensed into a 96-well culture plate by 20 μL, and then 10% fetal bovine serum (Luc2CP-HeLa cells suspended in a minimum essential medium (MEM) containing FBS and SAFC Biosciences (SAFC Biosciences) were seeded at a cell number of 7500/80 μL / well, 37 ° C, 5% Each formulation was introduced into Luc2CP-HeLa cells by culturing under CO ₂ conditions, and cells that were not treated as a negative control group were seeded.
Cells introduced with each preparation are cultured in a 5% CO ₂ incubator at 37 ° C for 24 hours, and a cell proliferation test assay (CellTiter-Fluor Cell Viability Assay, Promega, G6080) is used. After processing according to the method described in, and measuring the fluorescence intensity with a plate reader, using the luciferase quantification system (Steady-Glo Luciferase Assay System, Promega, E2520), the method described in the instructions attached to the product Then, the luminescence intensity was measured with a plate reader. The amount of luciferin luminescence obtained was corrected with the amount of fluorescence obtained in the cell proliferation test assay. The amount of luminescence of each preparation-treated group was calculated as a relative ratio when the corrected luminescence amount of the negative control group was 1.
As is apparent from Table 28, the expression rate of Luc after introducing Formulations 1 to 21 containing II-25 as lipid A into the human cervical cancer-derived cell line Luc2CP-HeLa cells is II-25-free. As a result, the inhibition rate was higher than those of preparations 22 to 42. In addition, the preparation containing II-25 as lipid A was not limited to a specific lipid B, and the inhibition rate was high.
Therefore, it has been clarified that the lipid nanoparticles containing lipid A of the present invention can introduce nucleic acid into cells and the like, and are preparations that facilitate drug delivery into cells in vitro.

II-35 obtained in Example 44, CL-1 obtained in Reference Example 1 and 1,2-dimyristoyl-sn-glycero-3-phosphoethanolamine-N- [methoxy (polyethylene glycol) -2000] Using (PEG-DMPE), 1,2-distearoyl-sn-glycero-3-phosphocholine (DSPC) and cholesterol, a preparation 43 containing nucleic acid-containing lipid nanoparticles was produced as follows.
The nucleic acid used is a siRNA that suppresses the expression of a luciferase (hereinafter referred to as Luc) gene consisting of a base sequence of a sense strand (5′-CCGUCGUAAUUCGUGACACAGA-3 ′) and an antisense strand (5′-UUGCUCACGAAUACGACGGUG-3 ′). Yes, obtained from GeneDesign (hereinafter "Luc siRNA"). PEG-DMPE, DSPC, and cholesterol were obtained from NOF.
II-35 was dissolved in 100% ethanol to give 5 mg / mL to obtain a lipid stock solution. CL-1, PEG-DMPE, DSPC, and cholesterol were dissolved in 100% ethanol so that each would be 20 mg / mL to obtain a lipid stock solution. Each lipid stock solution was stored at −20 ° C. and heated to 60 ° C. immediately before preparation preparation to dissolve the lipid and then returned to room temperature before use.
Luc siRNA was dissolved in water for injection at 1 mg / mL to give a Luc siRNA solution.
II-35 as the lipid stock solution was added to 20 mL of 80% ethanol aqueous solution so as to be 0.313 μmol. Subsequently, 200 μL of the Luc siRNA solution was added and stirred for 1 minute, and then CL-1 / PEG-DMPE / DSPC / Chol as the lipid stock solution was added to the solution as 1.88 μmol / 0.145 μmol / 0.515 μmol / 1.07 μmol. It added so that it might become. Thereafter, water for injection was added at a flow rate of 62 mL / second or more to form a 20% or less aqueous ethanol solution to form a crude preparation. The obtained crude preparation was concentrated using Amicon Ultra (manufactured by Millipore), further replacing the solvent with physiological saline, and filtered in a clean bench using a 0.2 μm filter (manufactured by Toyo Roshi Kaisha, Ltd.). Further, the siRNA concentration of the obtained preparation was measured, and diluted with physiological saline so that the siRNA concentration was 0.1 mg / mL, whereby preparation 43 was obtained.

Preparation 44 was obtained in the same manner as in Example 93 except that CL-1 of preparation 43 was changed to CL-2 obtained in Reference Example 2.

Preparation 45 was obtained in the same manner as in Example 93 except that CL-1 in preparation 43 was changed to CL-3 obtained in Reference Example 3.

Preparation 46 was obtained in the same manner as in Example 93 except that CL-1 of preparation 43 was changed to CL-4 obtained in Reference Example 4.

Preparation 47 was obtained in the same manner as in Example 93 except that CL-1 of preparation 43 was changed to CL-5 obtained in Reference Example 5.

Preparation 48 was obtained in the same manner as in Example 93 except that CL-1 of preparation 43 was changed to CL-6 obtained in Reference Example 6.

Preparation 49 was obtained in the same manner as in Example 93 except that CL-1 of preparation 43 was changed to CL-7 obtained in Reference Example 7.

Preparation 50 was obtained in the same manner as in Example 93 except that CL-1 of preparation 43 was changed to CL-8 obtained in Reference Example 8.

Preparation 51 was obtained in the same manner as in Example 93 except that CL-1 of preparation 43 was changed to CL-9 obtained in Reference Example 9.

Preparation 52 was obtained in the same manner as in Example 93 except that CL-1 of preparation 43 was changed to CL-10 obtained in Reference Example 10.

Preparation 53 was obtained in the same manner as in Example 93, except that CL-1 of preparation 43 was changed to CL-11 obtained in Reference Example 11.

Preparation 54 was obtained in the same manner as in Example 93 except that CL-1 of preparation 43 was changed to CL-12 obtained in Reference Example 12.

Preparation 55 was obtained in the same manner as in Example 93 except that CL-1 of preparation 43 was changed to CL-13 obtained in Reference Example 13.

Test example 3
Measurement of average particle size of nucleic acid-containing lipid nanoparticles The average particle size of nucleic acid-containing lipid nanoparticles in the preparation was measured with a particle size measuring device (Zetasizer Nano ZS, manufactured by Malvern) (Table 29). PDI in the table represents a polydispersity index.

As a result, lipid A (II-35) and lipid B (CL-1-13), PEG-DMPE®, DSPC, a complex of cholesterol and nucleic acid was dispersed in ethanol, and water was rapidly added to the dispersion. Formulations 43 to 55 described in Examples 93 to 105 were formed with a small average particle size of 50 nm or less.

Test example 4
In vitro activity evaluation test of nucleic acid-containing lipid nanoparticles Formulations 43 to 55 described in Examples 93 to 105 and formulations 22 to 42 described in Comparative Examples 1 to 21 were forcibly expressed by the following methods, respectively. It was introduced into a cervical cancer-derived cell line HeLa cell (Luc2CP-HeLa).
Each formulation diluted with Optimem (Opti-MEM, Gibco) to a final nucleic acid concentration of 0.3 to 10 nM is dispensed into a 96-well culture plate by 20 μL, and then 10% fetal bovine serum (Luc2CP-HeLa cells suspended in a minimum essential medium (MEM) containing FBS and SAFC Biosciences (SAFC Biosciences) were seeded at a cell number of 7500/80 μL / well, 37 ° C, 5% Each formulation was introduced into Luc2CP-HeLa cells by culturing under CO ₂ conditions, and cells that were not treated as a negative control group were seeded.
Cells introduced with each preparation are cultured in a 5% CO ₂ incubator at 37 ° C for 24 hours, and a cell proliferation test assay (CellTiter-Fluor Cell Viability Assay, Promega, G6080) is used. After processing according to the method described in, and measuring the fluorescence intensity with a plate reader, using the luciferase quantification system (Steady-Glo Luciferase Assay System, Promega, E2520), the method described in the instructions attached to the product Then, the luminescence intensity was measured with a plate reader. The amount of luciferin luminescence obtained was corrected with the amount of fluorescence obtained in the cell proliferation test assay. The amount of luminescence of each preparation-treated group was calculated as a relative ratio when the corrected luminescence amount of the negative control group was 1.
As is apparent from Table 30, the expression rate of Luc after introducing preparations 43 to 55 containing II-35 as lipid A into human cervical cancer-derived cell line Luc2CP-HeLa cells is II-35-free. As a result, the inhibition rate was higher than those of preparations 22 to 42. Further, the preparation containing II-35 as lipid A was not limited to a specific lipid B, and the inhibition rate was high.
Therefore, it has been clarified that the lipid nanoparticles containing lipid A of the present invention can introduce nucleic acid into cells and the like, and are preparations that facilitate drug delivery into cells in vitro.

II-25 obtained in Example 34, CL-22 obtained in Reference Example 22 and 1,2-dimyristoyl-sn-glycero-3-phosphoethanolamine-N- [methoxy (polyethylene glycol) -2000] Formulation 56 containing nucleic acid-containing lipid nanoparticles was produced using (PEG-DMPE), 1,2-distearoyl-sn-glycero-3-phosphocholine (DSPC), and cholesterol as follows.
The nucleic acid used is a siRNA that suppresses the expression of a luciferase (hereinafter referred to as Luc) gene consisting of a base sequence of a sense strand (5′-CCGUCGUAAUUCGUGACACAGA-3 ′) and an antisense strand (5′-UUGCUCACGAAUACGACGGUG-3 ′). Yes, obtained from GeneDesign (hereinafter "Luc siRNA"). PEG-DMPE, DSPC, and cholesterol were obtained from NOF.
II-25 was dissolved in 100% ethanol so as to be 5 mg / mL each to prepare a lipid stock solution. CL-22, PEG-DMPE, DSPC, and cholesterol were dissolved in 100% ethanol so that each would be 20 mg / mL to obtain a lipid stock solution. Each lipid stock solution was stored at −20 ° C. and heated to 60 ° C. immediately before preparation preparation to dissolve the lipid and then returned to room temperature before use.
Luc siRNA was dissolved in water for injection at 1 mg / mL to give a Luc siRNA solution.
II-25 used as the lipid stock solution was added to 20 mL of an 80% aqueous ethanol solution to a concentration of 0.313 μmol. Subsequently, after adding 200 μL of the Luc siRNA solution and stirring for 1 minute, CL-22 / PEG-DMPE / DSPC / Chol as the lipid stock solution was added to this solution as 1.88 μmol / 0.145 μmol / 0.515 μmol / 1.07 μmol. It added so that it might become. Thereafter, water for injection was added at a flow rate of 62 mL / second or more to form a 20% or less aqueous ethanol solution to form a crude preparation. The obtained crude preparation was concentrated using Amicon Ultra (manufactured by Millipore), further replacing the solvent with physiological saline, and filtered in a clean bench using a 0.2 μm filter (manufactured by Toyo Roshi Kaisha, Ltd.). Further, the siRNA concentration of the obtained preparation was measured, and diluted with physiological saline so that the siRNA concentration was 0.1 mg / mL, whereby preparation 56 was obtained.

Preparation 57 was obtained in the same manner as in Example 106, except that CL-22 of preparation 56 was changed to CL-23 obtained in Reference Example 23.

Preparation 58 was obtained in the same manner as in Example 106 except that CL-22 of preparation 56 was changed to CL-24 obtained in Reference Example 24.

Preparation 59 was obtained in the same manner as in Example 106, except that CL-22 of preparation 56 was changed to CL-25 obtained in Reference Example 25.

Preparation 60 was obtained in the same manner as in Example 106, except that CL-22 of preparation 56 was changed to CL-26 obtained in Reference Example 26.

Preparation 61 was obtained in the same manner as in Example 106, except that CL-22 of Preparation 56 was changed to CL-27 obtained in Reference Example 27.

Preparation 62 was obtained in the same manner as in Example 106, except that CL-22 of Preparation 56 was changed to CL-28 obtained in Reference Example 28.

Preparation 63 was obtained in the same manner as in Example 106, except that CL-22 of Preparation 56 was changed to CL-29 obtained in Reference Example 29.

Preparation 64 was obtained in the same manner as in Example 106, except that CL-22 of preparation 56 was changed to CL-30 obtained in Reference Example 30.

Preparation 65 was obtained in the same manner as in Example 106, except that CL-22 of Preparation 56 was changed to CL-31 obtained in Reference Example 31.

Preparation 66 was obtained in the same manner as in Example 106, except that CL-22 of preparation 56 was changed to CL-32 obtained in Reference Example 32.

Preparation 67 was obtained in the same manner as Example 106, except that CL-22 of preparation 56 was changed to CL-33 obtained in Reference Example 33.

Preparation 68 was obtained in the same manner as in Example 106 except that CL-22 of preparation 56 was changed to CL-34 obtained in Reference Example 34.

Preparation 69 was obtained in the same manner as in Example 106 except that CL-22 of preparation 56 was changed to CL-35 obtained in Reference Example 35.

Preparation 70 was obtained in the same manner as in Example 106, except that CL-22 of Preparation 56 was changed to CL-36 obtained in Reference Example 36.

Preparation 71 was obtained in the same manner as in Example 106, except that CL-22 of Preparation 56 was changed to CL-37 obtained in Reference Example 37.

Preparation 72 was obtained in the same manner as in Example 106, except that CL-22 of Preparation 56 was changed to CL-38 obtained in Reference Example 38.

Preparation 73 was obtained in the same manner as in Example 106, except that CL-22 of Preparation 56 was changed to CL-39 obtained in Reference Example 39.

Preparation 74 was obtained in the same manner as in Example 106 except that CL-22 of preparation 56 was changed to CL-40 obtained in Reference Example 40.

Preparation 75 was obtained in the same manner as in Example 106, except that CL-22 of Preparation 56 was changed to CL-41 obtained in Reference Example 41.

Preparation 76 was obtained in the same manner as in Example 106, except that CL-22 of preparation 56 was changed to CL-42 obtained in Reference Example 42.

Preparation 77 was obtained in the same manner as in Example 106 except that CL-22 of preparation 56 was changed to CL-43 obtained in Reference Example 43.

Preparation 78 was obtained in the same manner as in Example 106, except that CL-22 of Preparation 56 was changed to CL-44 obtained in Reference Example 44.

Preparation 79 was obtained in the same manner as in Example 106, except that CL-22 of Preparation 56 was changed to CL-45 obtained in Reference Example 45.

II-35 obtained in Example 44, CL-22 obtained in Reference Example 22, and 1,2-dimyristoyl-sn-glycero-3-phosphoethanolamine-N- [methoxy (polyethylene glycol) -2000] Using 80 (PEG-DMPE), 1,2-distearoyl-sn-glycero-3-phosphocholine (DSPC) and cholesterol, a preparation 80 containing nucleic acid-containing lipid nanoparticles was produced as follows.
The nucleic acid used is a siRNA that suppresses the expression of a luciferase (hereinafter referred to as Luc) gene consisting of a base sequence of a sense strand (5′-CCGUCGUAAUUCGUGACACAGA-3 ′) and an antisense strand (5′-UUGCUCACGAAUACGACGGUG-3 ′). Yes, obtained from GeneDesign (hereinafter "Luc siRNA"). PEG-DMPE, DSPC, and cholesterol were obtained from NOF.
II-35 was dissolved in 100% ethanol so as to be 5 mg / mL each to prepare a lipid stock solution. CL-22, PEG-DMPE, DSPC, and cholesterol were dissolved in 100% ethanol so that each would be 20 mg / mL to obtain a lipid stock solution. Each lipid stock solution was stored at −20 ° C. and heated to 60 ° C. immediately before preparation preparation to dissolve the lipid and then returned to room temperature before use.
Luc siRNA was dissolved in water for injection at 1 mg / mL to give a Luc siRNA solution.
II-35 as the lipid stock solution was added to 20 mL of 80% ethanol aqueous solution so as to be 0.313 μmol. Subsequently, after adding 200 μL of the Luc siRNA solution and stirring for 1 minute, CL-22 / PEG-DMPE / DSPC / Chol as the lipid stock solution was added to this solution as 1.88 μmol / 0.145 μmol / 0.515 μmol / 1.07 μmol. It added so that it might become. Thereafter, water for injection was added at a flow rate of 62 mL / second or more to form a 20% or less aqueous ethanol solution to form a crude preparation. The obtained crude preparation was concentrated using Amicon Ultra (manufactured by Millipore), further replacing the solvent with physiological saline, and filtered in a clean bench using a 0.2 μm filter (manufactured by Toyo Roshi Kaisha, Ltd.). Further, the siRNA concentration of the obtained preparation was measured, and diluted with physiological saline so that the siRNA concentration was 0.1 mg / mL, whereby preparation 80 was obtained.

Preparation 81 was obtained in the same manner as Example 130, except that CL-22 of preparation 80 was changed to CL-23 obtained in Reference Example 23.

Test Example 5
Measurement of average particle size of nucleic acid-containing lipid nanoparticles The average particle size of nucleic acid-containing lipid nanoparticles in the preparation was measured with a particle size measurement device (Zetasizer Nano ZS, manufactured by Malvern) (Table 31). PDI in the table represents a polydispersity index.

As a result, lipid A (II-25, II-35) and lipid B (CL-22-45), PEG-DMPE, DSPC, and a complex of cholesterol and nucleic acid are dispersed in ethanol and rapidly dispersed into the dispersion. Formulations 56 to 81 described in Examples 106 to 131 formed by adding water had a small average particle size and was 50 nm or less.

Test Example 6
In Vitro Activity Evaluation Test of Nucleic Acid-Containing Lipid Nanoparticles Human Cervical Cancer-Derived Cell Line HeLa Cells (Luc2CP-HeLa) Introduced.
Each formulation diluted with Optimem (Opti-MEM, Gibco) to a final nucleic acid concentration of 0.3 to 10 nM is dispensed into a 96-well culture plate by 20 μL, and then 10% fetal bovine serum (Luc2CP-HeLa cells suspended in a minimum essential medium (MEM) containing FBS and SAFC Biosciences (SAFC Biosciences) were seeded at a cell number of 7500/80 μL / well, 37 ° C, 5% Each formulation was introduced into Luc2CP-HeLa cells by culturing under CO ₂ conditions, and cells that were not treated as a negative control group were seeded.
Cells introduced with each preparation are cultured in a 5% CO ₂ incubator at 37 ° C for 24 hours, and a cell proliferation test assay (CellTiter-Fluor Cell Viability Assay, Promega, G6080) is used. After processing according to the method described in, and measuring the fluorescence intensity with a plate reader, using the luciferase quantification system (Steady-Glo Luciferase Assay System, Promega, E2520), the method described in the instructions attached to the product Then, the luminescence intensity was measured with a plate reader. The amount of luciferin luminescence obtained was corrected with the amount of fluorescence obtained in the cell proliferation test assay. The amount of luminescence of each preparation-treated group was calculated as a relative ratio when the corrected luminescence amount of the negative control group was 1.
As is clear from Table 32, preparations containing lipid A (formulations 56 to 81) suppressed the expression of Luc regardless of lipid B.
Therefore, it has been clarified that the lipid nanoparticles containing lipid A of the present invention can introduce nucleic acid into cells and the like, and are preparations that facilitate drug delivery into cells in vitro.

II-25 obtained in Example 34, CL-46 obtained in Reference Example 46 and 1,2-dimyristoyl-sn-glycero-3-phosphoethanolamine-N- [methoxy (polyethylene glycol) -2000] A preparation 82 containing nucleic acid-containing lipid nanoparticles was produced using (PEG-DMPE), 1,2-distearoyl-sn-glycero-3-phosphocholine (DSPC), and cholesterol as follows.
The nucleic acid used is a siRNA that suppresses the expression of a luciferase (hereinafter referred to as Luc) gene consisting of a base sequence of a sense strand (5′-CCGUCGUAAUUCGUGACACAGA-3 ′) and an antisense strand (5′-UUGCUCACGAAUACGACGGUG-3 ′). Yes, obtained from GeneDesign (hereinafter "Luc siRNA"). PEG-DMPE, DSPC, and cholesterol were obtained from NOF.
II-25 was dissolved in 100% ethanol so as to be 5 mg / mL each to prepare a lipid stock solution. CL-46, PEG-DMPE, DSPC, and cholesterol were dissolved in 100% ethanol so as to be 20 mg / mL, respectively, to obtain a lipid stock solution. Each lipid stock solution was stored at −20 ° C. and heated to 60 ° C. immediately before preparation preparation to dissolve the lipid and then returned to room temperature before use.
Luc siRNA was dissolved in water for injection at 1 mg / mL to give a Luc siRNA solution.
II-25 used as the lipid stock solution was added to 20 mL of an 80% aqueous ethanol solution to a concentration of 0.313 μmol. Subsequently, after adding 200 μL of the Luc siRNA solution and stirring for 1 minute, CL-46 / PEG-DMPE / DSPC / Chol as the lipid stock solution was added to the solution as 1.88 μmol / 0.145 μmol / 0.515 μmol / 1.07 μmol. It added so that it might become. Thereafter, water for injection was added at a flow rate of 62 mL / second or more to form a 20% or less aqueous ethanol solution to form a crude preparation. The obtained crude preparation was concentrated using Amicon Ultra (manufactured by Millipore), further replacing the solvent with physiological saline, and filtered in a clean bench using a 0.2 μm filter (manufactured by Toyo Roshi Kaisha, Ltd.). Further, the siRNA concentration of the obtained preparation was measured, and diluted with physiological saline so that the siRNA concentration was 0.1 mg / mL, whereby preparation 82 was obtained.

Preparation 83 was obtained in the same manner as in Example 132, except that CL-46 of preparation 82 was changed to CL-47 obtained in Reference Example 47.

Preparation 84 was obtained in the same manner as in Example 132, except that CL-46 of preparation 82 was changed to CL-48 obtained in Reference Example 48.

Preparation 85 was obtained in the same manner as Example 132, except that CL-46 of preparation 82 was changed to CL-49 obtained in Reference Example 49.

Preparation 86 was obtained in the same manner as in Example 132, except that CL-46 of preparation 82 was changed to CL-50 obtained in Reference Example 50.

II-35 obtained in Example 44, CL-21 obtained in Reference Example 21 and 1,2-dimyristoyl-sn-glycero-3-phosphoethanolamine-N- [methoxy (polyethylene glycol) -2000] Using (PEG-DMPE), 1,2-distearoyl-sn-glycero-3-phosphocholine (DSPC) and cholesterol, a preparation 87 containing nucleic acid-containing lipid nanoparticles was produced as follows.
The nucleic acid used is a siRNA that suppresses the expression of a luciferase (hereinafter referred to as Luc) gene consisting of a base sequence of a sense strand (5′-CCGUCGUAAUUCGUGACACAGA-3 ′) and an antisense strand (5′-UUGCUCACGAAUACGACGGUG-3 ′). Yes, obtained from GeneDesign (hereinafter "Luc siRNA"). PEG-DMPE, DSPC, and cholesterol were obtained from NOF.
II-35 was dissolved in 100% ethanol so as to be 5 mg / mL each to prepare a lipid stock solution. CL-21, PEG-DMPE, DSPC, and cholesterol were dissolved in 100% ethanol so that each would be 20 mg / mL to obtain a lipid stock solution. Each lipid stock solution was stored at −20 ° C. and heated to 60 ° C. immediately before preparation preparation to dissolve the lipid and then returned to room temperature before use.
Luc siRNA was dissolved in water for injection at 1 mg / mL to give a Luc siRNA solution.
II-35 as the lipid stock solution was added to 20 mL of 80% ethanol aqueous solution so as to be 0.313 μmol. Subsequently, after adding 200 μL of the Luc siRNA solution and stirring for 1 minute, CL-21 / PEG-DMPE / DSPC / Chol as the lipid stock solution was added to this solution as 1.88 μmol / 0.145 μmol / 0.515 μmol / 1.07 μmol. It added so that it might become. Thereafter, water for injection was added at a flow rate of 62 mL / second or more to form a 20% or less aqueous ethanol solution to form a crude preparation. The obtained crude preparation was concentrated using Amicon Ultra (manufactured by Millipore), further replacing the solvent with physiological saline, and filtered in a clean bench using a 0.2 μm filter (manufactured by Toyo Roshi Kaisha, Ltd.). Further, the siRNA concentration of the obtained preparation was measured, and diluted with physiological saline so that the siRNA concentration was 0.1 mg / mL, whereby preparation 87 was obtained.

Preparation 88 was obtained in the same manner as in Example 137, except that CL-21 of preparation 87 was changed to CL-47 obtained in Reference Example 47.

Formulation 89 was obtained in the same manner as in Example 137, except that CL-21 of formulation 87 was changed to CL-48 obtained in Reference Example 48.

Preparation 90 was obtained in the same manner as in Example 137 except that CL-21 of preparation 87 was changed to CL-49 obtained in Reference Example 49.

II-3 obtained in Example 10, CL-13 obtained in Reference Example 13 and 1,2-dimyristoyl-sn-glycero-3-phosphoethanolamine-N- [methoxy (polyethylene glycol) -2000] Formulation 91 containing nucleic acid-containing lipid nanoparticles was produced using (PEG-DMPE), 1,2-distearoyl-sn-glycero-3-phosphocholine (DSPC), and cholesterol as follows.
The nucleic acid used is a siRNA that suppresses the expression of a luciferase (hereinafter referred to as Luc) gene consisting of a base sequence of a sense strand (5′-CCGUCGUAAUUCGUGACACAGA-3 ′) and an antisense strand (5′-UUGCUCACGAAUACGACGGUG-3 ′). Yes, obtained from GeneDesign (hereinafter "Luc siRNA"). PEG-DMPE, DSPC, and cholesterol were obtained from NOF.
II-3 was dissolved in 100% ethanol so as to be 5 mg / mL each to prepare a lipid stock solution. CL-13, PEG-DMPE, DSPC, and cholesterol were dissolved in 100% ethanol so as to be 20 mg / mL, respectively, to obtain a lipid stock solution. Each lipid stock solution was stored at −20 ° C. and heated to 60 ° C. immediately before preparation preparation to dissolve the lipid and then returned to room temperature before use.
Luc siRNA was dissolved in water for injection at 1 mg / mL to prepare a Luc siRNA solution.
II-3 used as the lipid stock solution was added to 20 mL of 80% ethanol aqueous solution to a concentration of 0.313 μmol. Subsequently, after adding 200 μL of the Luc siRNA solution and stirring for 1 minute, CL-13 / PEG-DMPE / DSPC / Chol as the lipid stock solution was added to the solution as 1.88 μmol / 0.145 μmol / 0.515 μmol / 1.07 μmol. It added so that it might become. Thereafter, water for injection was added at a flow rate of 62 mL / second or more to form a 20% or less aqueous ethanol solution to form a crude preparation. The obtained crude preparation was concentrated using Amicon Ultra (manufactured by Millipore), further replacing the solvent with physiological saline, and filtered in a clean bench using a 0.2 μm filter (manufactured by Toyo Roshi Kaisha, Ltd.). Furthermore, the siRNA concentration of the obtained preparation was measured, and diluted with physiological saline so that the siRNA concentration was 0.1 mg / mL, to obtain a preparation 91.

Preparation 92 was obtained in the same manner as in Example 141 except that CL-13 in Preparation 91 was changed to CL-50 obtained in Reference Example 50.

Test Example 7
Measurement of average particle size of nucleic acid-containing lipid nanoparticles The average particle size of nucleic acid-containing lipid nanoparticles in the preparation was measured with a particle size measurement device (Zetasizer Nano ZS, manufactured by Malvern) (Table 33). PDI in the table represents a polydispersity index.

As a result, lipid A (II-25, II-35, II-3) and various lipid B (CL-21, 46-50), PEG-DMPE, DSPC, and a complex of cholesterol and nucleic acid are dispersed in ethanol. The formulations 82 to 92 described in Examples 132 to 142, which were formed by rapidly adding water to the dispersion, had a small average particle size of 50 nm or less.

(R) -2-((2R, 3R, 4S) -3,4-bis (dodecanoyloxy) tetrahydrofuran-2-yl) -2- (dodecanoyloxy) -N, N, N-trimethylethane-1 -Aminium chloride (compound V'-2)
Process 1
It was synthesized by the method according to the method described in “European Journal of Organic Chemistry (Eur.J.Org.Chem.)”, 2013, Vol. 3, p.566-577 (2R, 3R, 4S ) -3,4-bis (benzyloxy) -2-((R) -1- (benzyloxy) -2- (trityloxy) ethyl) tetrahydrofuran (5.97 g, 8.82 mmol) dissolved in methanol (150 mL) Then, paratoluenesulfonic acid monohydrate (4.19 g, 22.1 mmol) was added, and the mixture was stirred at room temperature for 3.5 hours. The reaction solution was concentrated under reduced pressure, and saturated multistory water was added to the residue, followed by extraction with dichloromethane. The organic layer was dried over anhydrous sodium sulfate, filtered and concentrated under reduced pressure. The obtained residue was purified by silica gel column chromatography (heptane / ethyl acetate = 95 / 5-60 / 40) to give (R) -2- (benzyloxy) -2-((2R, 3R, 4S) -3 , 4-bis (benzyloxy) tetrahydrofuran-2-yl) ethan-1-ol (3.56 g, 8.19 mmol, 93.0% yield) was obtained.
ESI-MS m / z: 435 (M + H) ⁺ .
Process 2
(R) -2- (benzyloxy) -2-((2R, 3R, 4S) -3,4-bis (benzyloxy) tetrahydrofuran-2-yl) ethan-1-ol (1.53) obtained in step 1 g, 3.52 mmol) was dissolved in dichloromethane (65.0 mL), triethylamine (1.24 mL, 8.87 mmol) and methanesulfonyl chloride (0.550 mL, 7.04 mmol) were added under ice-cooling, and the mixture was stirred at the same temperature for 2.5 hours. Water was added to the reaction mixture, and the mixture was extracted with dichloromethane. The organic layer was dried over anhydrous sodium sulfate, filtered, concentrated under reduced pressure, and (R) -2- (benzyloxy) -2-((2R, 3R, 4S) -3,4-bis (benzyloxy) tetrahydrofuran. -2-yl) ethyl methanesulfonate (1.91 g) was obtained.
ESI-MS m / z: 513 (M + H) ⁺ .
(R) -2- (Benzyloxy) -2-((2R, 3R, 4S) -3,4-bis (benzyloxy) tetrahydrofuran-2-yl) ethyl methanesulfonate (0.805 g, 1.57 mmol) It was dissolved in chloroform (4.00 mL), a dimethylamine solution (15.6 mL, 31.2 mmol, 2M in THF) was added, and the mixture was stirred at 130 ° C. for 7.5 hours under microwave irradiation. The reaction mixture was concentrated under reduced pressure, and the resulting residue was purified by silica gel column chromatography (chloroform / methanol = 100/0 to 90/10) to give (R) -2- (benzyloxy) -2-((2R , 3R, 4S) -3,4-bis (benzyloxy) tetrahydrofuran-2-yl) -N, N-dimethylethan-1-amine (0.555 g, 1.20 mmol, yield 77.0%) was obtained.
ESI-MS m / z: 462 (M + H) ⁺ .
Process 3
(R) -2- (Benzyloxy) -2-((2R, 3R, 4S) -3,4-bis (benzyloxy) tetrahydrofuran-2-yl) -N, N-dimethylethane obtained in Step 2 -1-amine (0.774 g, 1.68 mmol) was dissolved in ethanol (34.0 mL), carbon-supported palladium hydroxide (1.56 g, 20 wt% dry basis, <50% water), acetic acid (0.590 mL) were added, After purging with hydrogen, the mixture was stirred overnight. The reaction solution was filtered through Celite while washing with ethanol, and the filtrate was concentrated under reduced pressure to obtain (2R, 3R, 4S) -2-((R) -2- (dimethylamino) -1-hydroxyethyl) tetrahydrofuran-3. , 4-diol (0.407 g, 1.62 mmol, 97.0% yield) was obtained.
ESI-MS m / z: 192 (M + H) ⁺ .
Process 4
The (2R, 3R, 4S) -2-((R) -2- (dimethylamino) -1-hydroxyethyl) tetrahydrofuran-3,4-diol (0.141 g, 0.560 mmol) obtained in Step 3 was converted to dimethylformamide. (6.00 mL), triethylamine (0.660 mL, 4.75 mmol) and lauroyl chloride (0.900 mL, 3.80 mmol) were added, and the mixture was stirred at room temperature overnight. Triethylamine (0.250 mL, 1.80 mmol) and lauroyl chloride (0.150 mL, 0.634 mmol) were added, and the mixture was stirred at room temperature for 2 hours. Saturated aqueous sodium hydrogen carbonate was added to the reaction mixture, and the mixture was extracted with ethyl acetate. The organic layer was dried over anhydrous sodium sulfate, filtered and concentrated under reduced pressure. The obtained residue was purified by silica gel column chromatography (heptane / ethyl acetate = 100 / 0-80 / 20) to give (2R, 3R, 4S) -2-((R) -2- (dimethylamino) -1 -(Dodecanoyloxy) ethyl) tetrahydrofuran-3,4-diyldidodecanoate (0.215 g, 0.291 mmol, yield 52.0%) was obtained.
ESI-MS m / z: 739 (M + H) ⁺ .
Process 5
(2R, 3R, 4S) -2-((R) -2- (dimethylamino) -1- (dodecanoyloxy) ethyl) tetrahydrofuran-3,4-diyldidodecanoate (0.203 g, 0.276 mmol) was used in the same manner as in Step 2 of Example 8 to obtain the title compound (0.0855 g, 0.108 mmol, yield 40.3%).
ESI-MS m / z: 753 (M) ⁺ ; ¹ H-NMR (CDCl ₃ ) δ: 0.88 (t, J = 6.8 Hz, 9H), 1.21-1.37 (m, 48H), 1.49-1.67 (m, 6H), 2.29-2.37 (m, 6H), 3.49 (s, 9H), 3.71-3.80 (m, 2H), 4.12 (dd, J = 9.0, 3.3 Hz, 1H), 4.25 (dd, J = 10.6, 4.6 Hz, 1H), 4.51 (dd, J = 14.7, 9.0 Hz, 1H), 5.04 (d, J = 4.6 Hz, 1H), 5.30 (d, J = 3.3 Hz, 1H), 5.50 (t, J = 9.0 Hz, 1H).

(R) -2-((2R, 3R, 4S) -3,4-bis (tetradecanoyloxy) tetrahydrofuran-2-yl) -N, N, N-trimethyl-2- (tetradecanoyloxy) ethane -1-Aminium chloride (Compound V'-3)
Process 1
Example 143 Step 4 of (2R, 3R, 4S) -2-((R) -2- (dimethylamino) -1-hydroxyethyl) tetrahydrofuran-3,4-diol obtained in Step 3 In the same manner, using myristoyl chloride instead of lauroyl chloride, (2R, 3R, 4S) -2-((R) -2- (dimethylamino) -1- (tetradecanoyloxy) ethyl) tetrahydrofuran- 3,4-Diylditetradecanoate (0.164 g, 0.200 mmol, 55.0%) was obtained.
ESI-MS m / z: 823 (M + H) ⁺ .
Process 2
(2R, 3R, 4S) -2-((R) -2- (dimethylamino) -1- (tetradecanoyloxy) ethyl) tetrahydrofuran-3,4-diylditetradecanoate obtained in step 1 (0.109 g, 0.132 mmol) was used in the same manner as in Step 2 of Example 8 to obtain the title compound (0.0701 g, 0.0800 mmol, yield 60.9%).
ESI-MS m / z: 837 (M) ⁺ ; ¹ H-NMR (CDCl ₃ ) δ: 0.88 (t, J = 6.8 Hz, 9H), 1.20-1.35 (m, 60H), 1.49-1.67 (m, 6H), 2.29-2.37 (m, 6H), 3.44 (s, 9H), 3.73-3.81 (m, 2H), 4.11 (dd, J = 9.3, 2.9 Hz, 1H), 4.25 (dd, J = 10.5, 4.4 Hz, 1H), 4.31-4.33 (m, 1H), 5.04 (d, J = 4.4 Hz, 1H), 5.29 (d, J = 2.9 Hz, 1H), 5.49 (t, J = 9.3 Hz, 1H) .

(R) -2-((2R, 3R, 4S) -3,4-bis (palmitoyloxy) tetrahydrofuran-2-yl) -N, N, N-trimethyl-2- (palmitoyloxy) ethane-1-amino Nitric chloride (Compound V'-4)
Process 1
Example 143 Step 4 of (2R, 3R, 4S) -2-((R) -2- (dimethylamino) -1-hydroxyethyl) tetrahydrofuran-3,4-diol obtained in Step 3 In the same manner, using palmitoyl chloride instead of lauroyl chloride, (2R, 3R, 4S) -2-((R) -2- (dimethylamino) -1- (palmitoyloxy) ethyl) tetrahydrofuran-3, 4-Diyldipalmitate (0.0527 g, 0.0581 mmol, 11.7%) was obtained.
ESI-MS m / z: 907 (M + H) ⁺ .
Process 2
(2R, 3R, 4S) -2-((R) -2- (dimethylamino) -1- (palmitoyloxy) ethyl) tetrahydrofuran-3,4-diyldipalmitate (0.216 g, obtained in step 1) The title compound (0.0751 g, 0.0785 mmol, yield 33.0%) was obtained in the same manner as in Step 2 of Example 8 using 0.238 mmol).
ESI-MS m / z: 921 (M) ⁺ ; ¹ H-NMR (CDCl ₃ ) δ: 0.88 (t, J = 6.8 Hz, 9H), 1.22-1.35 (m, 72H), 1.49-1.66 (m, 6H), 2.29-2.37 (m, 6H), 3.42 (s, 9H), 3.74-3.80 (m, 2H), 4.11 (dd, J = 9.3, 3.5 Hz, 1H), 4.26 (dd, J = 10.9, 4.3 Hz, 1H), 4.35 (dd, J = 14.6, 8.0 Hz, 1H), 5.04 (d, J = 4.3 Hz, 1H), 5.30 (d, J = 3.5 Hz, 1H), 5.49 (t, J = 8.0, 1H).

(R) -2-((2R, 3R, 4S) -3,4-bis (stearoyloxy) tetrahydrofuran-2-yl) -N, N, N-trimethyl-2- (stearoyloxy) ethane-1-amino Nitric chloride (Compound V'-5)
Process 1
Example 143 Step 4 of (2R, 3R, 4S) -2-((R) -2- (dimethylamino) -1-hydroxyethyl) tetrahydrofuran-3,4-diol obtained in Step 3 In the same manner, using stearyl chloride instead of lauroyl chloride, (2R, 3R, 4S) -2-((R) -2- (dimethylamino) -1- (stearoyloxy) ethyl) tetrahydrofuran-3, 4-Diyl distearate (0.261 g, 0.263 mmol, 50.0%) was obtained.
ESI-MS m / z: 991 (M + H) ⁺ .
Process 2
(2R, 3R, 4S) -2-((R) -2- (Dimethylamino) -1- (stearoyloxy) ethyl) tetrahydrofuran-3,4-diyl distearate obtained in Step 1 (0.149 g, The title compound (0.144 g, 0.138 mmol, yield 92.0%) was obtained in the same manner as in Step 2 of Example 8 using 0.150 mmol).
ESI-MS m / z: 1005 (M) ⁺ ; ¹ H-NMR (CDCl ₃ ) δ: 0.87 (t, J = 6.8 Hz, 9H), 1.16-1.37 (m, 84H), 1.47-1.66 (m, 6H), 2.24-2.38 (m, 6H), 3.33 (s, 9H), 3.73-3.81 (m, 2H), 4.00-4.10 (m, 2H), 4.25 (dd, J = 10.8, 4.3 Hz, 1H) , 5.03 (d, J = 4.3 Hz, 1H), 5.28 (d, J = 3.0 Hz, 1H), 5.47 (t, J = 8.4 Hz, 1H).

V′-3 obtained in Example 144, CL-2 obtained in Reference Example 2 and 1,2-dimyristoyl-sn-glycero-3-phosphoethanolamine-N- [methoxy (polyethylene glycol) -2000 ] (PEG-DMPE), 1,2-distearoyl-sn-glycero-3-phosphocholine (DSPC), and cholesterol were used to produce Formulation 93 containing nucleic acid-containing lipid nanoparticles as follows.
The nucleic acid used is a siRNA that suppresses the expression of a luciferase (hereinafter referred to as Luc) gene consisting of a base sequence of a sense strand (5′-CCGUCGUAAUUCGUGACACAGA-3 ′) and an antisense strand (5′-UUGCUCACGAAUACGACGGUG-3 ′). Yes, obtained from GeneDesign (hereinafter "Luc siRNA"). PEG-DMPE, DSPC, and cholesterol were obtained from NOF.
V′-3 was dissolved in 100% ethanol so as to be 5 mg / mL to obtain a lipid stock solution. CL-2, PEG-DMPE, DSPC, and cholesterol were dissolved in 100% ethanol so that each would be 20 mg / mL to obtain a lipid stock solution. Each lipid stock solution was stored at −20 ° C. and heated to 60 ° C. immediately before preparation preparation to dissolve the lipid and then returned to room temperature before use.
Luc siRNA was dissolved in water for injection at 1 mg / mL to give a Luc siRNA solution.
V'-3 serving as the lipid stock solution was added to 20 mL of 80% ethanol aqueous solution so as to be 0.313 μmol. Subsequently, after adding 200 μL of the Luc siRNA solution and stirring for 1 minute, CL-2 / PEG-DMPE / DSPC / Chol as the lipid stock solution was added to this solution as 1.88 μmol / 0.145 μmol / 0.515 μmol / 1.07 μmol. It added so that it might become. Thereafter, water for injection was added at a flow rate of 62 mL / second or more to form a 20% or less aqueous ethanol solution to form a crude preparation. The obtained crude preparation was concentrated using Amicon Ultra (manufactured by Millipore), further replacing the solvent with physiological saline, and filtered in a clean bench using a 0.2 μm filter (manufactured by Toyo Roshi Kaisha, Ltd.). Further, the siRNA concentration of the obtained preparation was measured, and diluted with physiological saline so that the siRNA concentration was 0.1 mg / mL, whereby preparation 93 was obtained.

Preparation 94 was obtained in the same manner as in Example 147, except that CL-2 of preparation 93 was changed to CL-8 obtained in Reference Example 8.

Preparation 95 was obtained in the same manner as in Example 147, except that V′-3 of preparation 93 was changed to V′-5 obtained in Example 146.

A preparation 96 was obtained in the same manner as in Example 149, except that V′-3 of the preparation 94 was changed to V′-5 obtained in Example 146.

Preparation 97 was obtained in the same manner as in Example 147, except that V′-3 of Preparation 93 was changed to V′-2 obtained in Example 143.

A preparation 98 was obtained in the same manner as in Example 149, except that V′-3 of the preparation 94 was changed to V′-2 obtained in Example 143.

Preparation 99 was obtained in the same manner as in Example 147, except that V′-3 of Preparation 93 was changed to V′-4 obtained in Example 145.

Formulation 100 was obtained in the same manner as in Example 149, except that V′-3 of Formulation 94 was changed to V′-4 obtained in Example 145.

Formulation 101 was obtained in the same manner as in Example 72 except that CL-1 of Formulation 1 was changed to CL-53 obtained in Reference Example 53.

Preparation 102 was obtained in the same manner as in Example 72 except that CL-1 in preparation 1 was changed to CL-54 obtained in Reference Example 54.

Preparation 103 was obtained in the same manner as in Example 72, except that CL-1 of preparation 1 was changed to CL-55 obtained in Reference Example 55.

Formulation 104 was obtained in the same manner as in Example 72, except that CL-1 of Formulation 1 was changed to CL-56 obtained in Reference Example 56.

Formulation 105 was obtained in the same manner as in Example 72 except that CL-1 of Formulation 1 was changed to CL-57 obtained in Reference Example 57.

Preparation 106 was obtained in the same manner as in Example 72 except that CL-1 in preparation 1 was changed to CL-58 obtained in Reference Example 58.

Preparation 107 was obtained in the same manner as in Example 72 except that CL-1 in preparation 1 was changed to CL-59 obtained in Reference Example 59.

Preparation 108 was obtained in the same manner as in Example 72 except that CL-1 of preparation 1 was changed to CL-60 obtained in Reference Example 60.

Preparation 109 was obtained in the same manner as in Example 72, except that CL-1 in Preparation 1 was changed to CL-61 obtained in Reference Example 61.

Preparation 110 was obtained in the same manner as in Example 72 except that CL-1 of preparation 1 was changed to CL-62 obtained in Reference Example 62.

Formulation 111 was obtained in the same manner as in Example 72 except that CL-1 of formulation 1 was changed to CL-63 obtained in Reference Example 63.

Formulation 112 was obtained in the same manner as in Example 72 except that CL-1 of Formulation 1 was changed to CL-64 obtained in Reference Example 64.

Preparation 113 was obtained in the same manner as in Example 72 except that CL-1 in preparation 1 was changed to CL-65 obtained in Reference Example 65.

Preparation 114 was obtained in the same manner as in Example 141, except that CL-13 in Preparation 91 was changed to CL-54 obtained in Reference Example 54.

Preparation 115 was obtained in the same manner as in Example 141, except that CL-13 in Preparation 91 was changed to CL-55 obtained in Reference Example 54.

Test Example 8
Measurement of average particle size of nucleic acid-containing lipid nanoparticles The average particle size of nucleic acid-containing lipid nanoparticles in the preparation was measured with a particle size measurement device (Zetasizer Nano ZS, manufactured by Malvern) (Table 34). PDI in the table represents a polydispersity index.

As a result, lipid A (V'-2, V'-3, V'-4, V'-5, II-25, II-3) and various lipid B (CL-2, 8, 53-65) Formulations 93 to 115 described in Examples 147 to 169 formed by dispersing a complex of PEG-DMPE®, DSPC, cholesterol and nucleic acid in ethanol and rapidly adding water to the dispersion have an average particle size. Small, 50 nm or less.

Test Example 9
In Vitro Activity Evaluation Test of Nucleic Acid-Containing Lipid Nanoparticles Human cervical cancer-derived cell line HeLa cells (Luc2CP-HeLa) in which the preparations 93 to 115 described in Examples 147 to 169 were forcibly expressed by the following methods, respectively Introduced.
Each formulation diluted with Optimem (Opti-MEM, Gibco) to a final nucleic acid concentration of 0.3 to 10 nM is dispensed into a 96-well culture plate by 20 μL, and then 10% fetal bovine serum (Luc2CP-HeLa cells suspended in a minimum essential medium (MEM) containing FBS and SAFC Biosciences (SAFC Biosciences) were seeded at a cell number of 7500/80 μL / well, 37 ° C, 5% Each formulation was introduced into Luc2CP-HeLa cells by culturing under CO ₂ conditions, and cells that were not treated as a negative control group were seeded.
Cells introduced with each preparation are cultured in a 5% CO ₂ incubator at 37 ° C for 24 hours, and a cell proliferation test assay (CellTiter-Fluor Cell Viability Assay, Promega, G6080) is used. After processing according to the method described in, and measuring the fluorescence intensity with a plate reader, using the luciferase quantification system (Steady-Glo Luciferase Assay System, Promega, E2520), the method described in the instructions attached to the product Then, the luminescence intensity was measured with a plate reader. The amount of luciferin luminescence obtained was corrected with the amount of fluorescence obtained in the cell proliferation test assay. The amount of luminescence of each preparation-treated group was calculated as a relative ratio when the corrected luminescence amount of the negative control group was 1.
As is clear from Table 35, preparations containing lipid A (formulations 93 to 115) suppressed the expression of Luc regardless of lipid B.
Therefore, it has been clarified that the lipid nanoparticles containing lipid A of the present invention can introduce nucleic acid into cells and the like, and are preparations that facilitate drug delivery into cells in vitro.

SEQ ID NO: 1 shows the base sequence of the Luc siRNA sense strand.
SEQ ID NO: 2 shows the nucleotide sequence of the Luc siRNA antisense strand.

Claims

A lipid having a hydrophilic part having one quaternary ammonium group and three independent hydrocarbon groups which may be substituted (lipid A), one amino group which may be substituted or one quaternary ammonium group Nucleic acid, including a lipid (lipid B), a lipid derivative or fatty acid derivative of a water-soluble polymer, and a nucleic acid containing a hydrophilic moiety having a hydrophobic moiety having two independent hydrocarbon groups that may be substituted in the same molecule Contains lipid nanoparticles.

2. The lipid nano particles according to claim 1, wherein the number of moles of the quaternary ammonium group in lipid A is 0.01 times or more the number of moles of phosphorus atoms in the nucleic acid.

The lipid nanoparticles according to claim 1 or 2, wherein the content of lipid B is at least 0.1 times the molar amount of the total lipid.

The number of moles of the amino group or quaternary ammonium group in the hydrophilic part in the lipid B is 0.01 times or more the mole number of the phosphorus atom in the nucleic acid according to any one of claims 1 to 3. Lipid nanoparticles.

Lipid B
Formula (CL-I)

(Where
R ¹¹³ and R ¹¹⁴ are the same or different, linear or branched optionally substituted C8-C24 alkyl, C8-C24 alkenyl or C8-C24 alkynyl,
R ¹¹⁵ is a hydrogen atom, hydroxy, optionally substituted C1-C4 alkyl, C1-C4 alkoxy or C1-C4 acyloxy,
X ¹⁰⁹ and X ¹¹⁰ are the same or different and are C1-C3 alkyl or taken together to form C2-C8 alkylene;
L ¹¹⁵ is -CO-O-, -O-CO-, -NHCO- or -CONH-
p ¹⁰⁶ is an integer from 0 to 3,
p ¹⁰⁷ is an integer from 1 to 4)
Formula (CL-VIII)

(Where
X ¹¹⁵ and X ¹¹⁶ are the same or different and are a hydrogen atom or C1-C3 alkyl,
L ¹¹⁸ and L ¹¹⁹ are the same or different and may be linear or branched C8-C24 alkylene or C8-C24 alkenylene,
M ¹⁰¹ and M ¹⁰² are the same or different, and -C = C-, -OC (O)-, -C (O) O-, -SC (O)-, -C (O) S-, -OC (S )-, -C (S) O-, -SS-, -C (R ^'' ) = N-, -N = C (R ^'' )-, -C (R ^'' ) = NO-, -ON = C (R ^'' )-, -N (R ^'' ) C (O)-, -C (O) N (R ^'' )-, -N (R ^'' ) C (S)-, -C (S) N (R ^'' )-, -N (R ^'' ) C (O) N (R ^''' )-, -N (R ^'' ) C (O) O-, -OC (O) Selected from the group consisting of N (R ^'' )-and -OC (O) O-
R ^'' and R ^''' are the same or different and are a hydrogen atom or C1-C3 alkyl;
R ¹¹⁸ and R ¹¹⁹ are the same or different and are linear or branched optionally substituted C1-C16 alkyl or C2-C16 alkenyl),
Formula (CL-X)

(Where
X ¹¹⁹ and X ¹²⁰ are the same or different and each represents a hydrogen atom, a linear or branched C1-C20 alkyl, C1-C20 alkenyl, C1-C20 alkynyl or C6-C20 acyl,
R ¹²² and R ¹²³ are the same or different, linear or branched optionally substituted C1-C30 alkyl, C2-C30 alkenyl or C2-C30 alkynyl,
p ¹¹² , p ¹¹³ and p ¹¹⁴ are the same or different and are 0 or any positive integer),
Formula (CL-XII)

(Where
R ¹²⁶ and R ¹²⁷ are the same or different and may be linear or branched substituted C8-C24 alkyl, C8-C24 alkenyl, C8-C24 alkynyl, C8-C24 heteroalkyl, C8-C24 heteroalkenyl or C8-C24 heteroalkynyl,
X ¹²³ is a hydrogen atom or an optionally substituted C1-C6 alkyl,
X ¹²⁴ is C1-C6 alkyl, substituted C1-C6 alkyl substituted with -NR ^4a R ^4b or optionally substituted C3-C7 heterocyclyl;
R ^4a and R ^4b are the same or different and are a hydrogen atom, -C (= NH) NH ₂ or an optionally substituted C1-C6 alkyl, or R ^4a and R ^4b are a nitrogen atom to which they are bonded. Taken together to form an optionally substituted C3-C7 heterocyclyl,
X ¹²³ and X ¹²⁴ together with the nitrogen atom to which they are attached may form an optionally substituted C3-C7 heterocyclyl;
However, X ¹²³ and X ¹²⁴ do not form imidazolyl, benzimidazolyl, or succinimidyl, and only one primary amine can be present on either of X ¹²³ and X ¹²⁴ , or any primary amines also not present on either one of X ¹²³ and X ^124, X ¹²³ and X ¹²⁴ is not substituted amides,
When R ¹²⁶ and R ¹²⁷ are C11 alkyl or C15 alkyl, X ¹²³ is not a hydrogen atom,
When R ¹²⁶ and R ¹²⁷ are C16 alkyl or C17 alkyl, R ¹²⁶ and R ¹²⁷ are not substituted with OH;
When R ¹²⁶ and R ¹²⁷ are C17 alkyl, X ¹²³ and X ¹²⁴ are not substituted with OH;
When R ¹²⁶ and R ¹²⁷ are C18 alkyl, X ¹²⁴ is not substituted with an optionally substituted imidazolyl)
Formula (CL-XIV)

(Where
X ¹²⁵ and X ¹²⁶ are the same or different hydrogen atoms, optionally substituted C1-C6 alkyl, heterocyclyl or polyamine, or X ¹²⁵ and X ¹²⁶ together with the nitrogen to which they are attached, In addition to nitrogen, it may form a 4-7 membered monocyclic heterocycle which may contain 1 or 2 additional heteroatoms selected from N, O and S;
R ¹³⁰ is a hydrogen atom or C1-C6 alkyl;
R ¹²⁸ and R ¹²⁹ are the same or different, linear or branched optionally substituted C4-C24 alkyl or C4-C24 alkenyl,
Y3 and Y4 represents an oxygen atom or CH ₂ identical or different,
p115 is an integer from 0 to 2),
Formula (CL-XV)

(Where
X ¹²⁷ and X ¹²⁸ are the same or different and are C1-C6 alkyl, C2-C6 alkenyl, C2-C6 alkynyl,
X ¹²⁷ and X ¹²⁸ together with the nitrogen atom to which they are attached form a heterocyclic ring having 1 to 2 nitrogen atoms;
L ¹²² is -C (O) O-, -OC (O)-, -C (O) N (X ¹³⁰ )-, -N (X ¹³⁰ ) C (O)-, -OC (O) O-, -OC (O) N (X ¹³⁰ )-, -N (X ¹³⁰ ) C (O) N (X ¹³⁰ )-, or -N (X ¹³⁰ ) C (O) O-
Each ^occurrence of X ¹³⁰ is independently a hydrogen atom or C1-C3 alkyl;
a is 1, 2, 3, 4, 5, or 6;
b is 0, 1, 2, or 3;
X ¹²⁹ is absent or is hydrogen or C1-C3 alkyl;
R ¹³¹ and R ¹³² are the same or different and are alkyl having 12 to 24 carbon atoms, alkenyl having 12 to 24 carbon atoms, or alkoxy having 12 to 24 carbon atoms having one or more biodegradable groups, The decomposable group is independently incorporated into the alkyl group having 12 to 24 carbon atoms, the alkenyl group having 12 to 24 carbon atoms, or the alkoxy group having 12 to 24 carbon atoms, or an alkyl group having 12 to 24 carbon atoms. , At the end of an alkenyl group having 12 to 24 carbon atoms or an alkoxy group having 12 to 24 carbon atoms,
The biodegradable group is incorporated as —C (O) O—, —OC (O) —, —C (O) N (X ¹³⁰ ) —, or —N (X ¹³⁰ ) C ( O)-, which is present at the terminal, is -C (O) O-C1-C4 alkyl, -OC (O) -C1-C4 alkyl, -C (O) N (X ¹³⁰ ) -C1-C4 Alkyl, or -N (X ¹³⁰ ) C (O) -C1-C4 alkyl,
R ¹³¹ and R ¹³² have at least 4 carbon atoms between the biodegradable group and a tertiary carbon atom marked with an asterisk (*)),
Formula (CL-XVI)

(Where
R ¹³³ and R ¹³⁴ are the same or different and are linear or branched C1-C9 alkyl, C2-C11 alkenyl or C2-C11 alkynyl;
L ¹²³ and L ¹²⁴ are the same or different and are linear C5-C18 alkylene or linear C5-C18 alkenylene, or form a heterocyclic ring with N bonded thereto,
L ¹²⁵ is a single bond or -C (O) O-
When L ¹²⁵ is -C (O) O-, -L ¹²⁴ -CO-OR ¹³⁴ is formed,
L ¹²⁷ is S or O,
L ¹²⁶ is a single bond, a linear or branched C1-C6 alkylene, or forms a heterocyclic ring with N bonded via -C (O)-,
L ¹²⁸ is a linear or branched C1-C6 alkylene,
X ¹³¹ and X ¹³² are the same or different and are hydrogen or linear or branched C1-C6 alkyl), or formula (CL-XVII)

(Where
L ¹³¹ is C2-C4 alkylene or —CH ₂ —S—CH ₂ CH ₂ —,
L ¹²⁹ and L ¹³⁰ are the same or different and are C1-C6 alkyl;
R ¹³⁵ and R ¹³⁶ are the same or different and are C10-C30 alkyl, C10-C30 alkenyl,
X ¹³³ and X ¹³⁴ are the same or different and are hydrogen, C1-C6 alkyl or —CH ₂ CH ₂ OH. The lipid nanoparticle according to any one of claims 1 to 4, which is a lipid represented by the following formula or a combination thereof.

6. Lipid nanoparticles according to any one of claims 1 to 5, having an average particle diameter of 20 to 65 nm.

A method for producing nucleic acid-containing lipid nanoparticles comprising:
(a) a lipid having a hydrophilic part having one quaternary ammonium group and three independent hydrocarbon groups optionally substituted in a mixed solvent of water and an organic solvent miscible with water (lipid A) A lipid having a hydrophilic part having one amino group which may be substituted or one quaternary ammonium group and a hydrophobic part having two independent hydrocarbon groups which may be substituted (lipid B) and a nucleic acid to prepare a first lipid solution containing lipid A and nucleic acid,
(b) adding a second lipid solution containing a lipid derivative or fatty acid derivative of a water-soluble polymer to the first lipid solution to prepare a third lipid solution; and
(c) The production method comprising a step of adding water or an aqueous buffer solution to the third lipid solution.

A method for producing nucleic acid-containing lipid nanoparticles comprising:
(a) a lipid having a hydrophilic part having one quaternary ammonium group and three independent hydrocarbon groups optionally substituted in a mixed solvent of water and an organic solvent miscible with water (lipid A) A lipid having a hydrophilic part having one amino group which may be substituted or one quaternary ammonium group and a hydrophobic part having two independent hydrocarbon groups which may be substituted (lipid A step of mixing B), a nucleic acid, and a lipid derivative or fatty acid derivative of a water-soluble polymer to prepare a first lipid solution containing the lipid and the nucleic acid; and
(b) The production method comprising a step of adding water or an aqueous buffer solution to the first lipid solution.

Lipid B has the formula (CL-I)

(Where
L ¹³¹ is C2-C4 alkylene or —CH ₂ —S—CH ₂ CH ₂ —,
L ¹²⁹ and L ¹³⁰ are the same or different and are C1-C6 alkyl;
R ¹³⁵ and R ¹³⁶ are the same or different and are C10-C30 alkyl, C10-C30 alkenyl,
X ¹³³ and X ¹³⁴ are the same or different and are hydrogen, C1-C6 alkyl or —CH ₂ CH ₂ OH. 9. The production method according to claim 7 or 8, which is a lipid represented by

A method for introducing a nucleic acid into a cell using the nucleic acid-containing lipid nanoparticles according to any one of claims 1 to 6.

11. The method according to claim 10, wherein the cell is a cell in a mammalian tumor or inflammation site.

12. The method according to claim 10 or 11, wherein the cell is a cell in a mammalian liver, stomach, lung, kidney, pancreas or spleen.

The method according to any one of claims 10 to 12, wherein the method of introducing into a cell is a method of introducing into a cell by intravenous administration or subcutaneous administration.

A method for treating cancer or inflammatory disease, comprising administering the nucleic acid-containing lipid nanoparticles according to any one of claims 1 to 6 to a mammal.

15. The method according to claim 14, wherein the method of administration is intravenous administration or subcutaneous administration.

A medicament comprising the nucleic acid-containing lipid nanoparticles according to any one of claims 1 to 6.

17. The medicine according to claim 16, which is for intravenous administration or subcutaneous administration.

A therapeutic agent for cancer or inflammatory disease comprising the nucleic acid-containing lipid nanoparticles according to any one of claims 1 to 6.

19. The therapeutic agent according to claim 18, which is for intravenous administration or subcutaneous administration.