Compound or salt thereof, lipid particles, and pharmaceutical composition

Abstract

The present invention addresses the problem of providing: a compound or a salt thereof which constitutes lipid particles that make it possible to achieve a high nucleic acid encapsulation rate and excellent nucleic acid delivery; and lipid particles and a pharmaceutical composition which use the same and make it possible to achieve a high nucleic acid encapsulation rate and excellent nucleic acid delivery. The present invention provides a compound represented by formula (1) or a salt thereof. In the formula, the symbols have the meanings as defined in the specification of the present application.

Description

Compound or salt thereof, lipid particle and pharmaceutical composition

[0001] The present invention relates to a compound or a salt thereof, as well as lipid particles and pharmaceutical compositions using the same.

[0002] Nucleic acid drugs have a clear mechanism of action against diseases and few side effects, and are expected to be next-generation pharmaceuticals. For example, nucleic acid drugs using siRNA (small interfering RNA) can inhibit the expression of target genes in a sequence-specific manner within cells. As a result, diseases and symptoms caused by the abnormal expression of specific genes or groups of genes can be alleviated or treated. In order to express the functions of these nucleic acids, it is necessary to deliver the nucleic acid drugs into cells.

[0003] Methods for efficiently delivering nucleic acids into cells include methods using viral vectors such as retroviruses and adenoviruses. While methods using viral vectors have high gene transfer efficiency, they are limited in the size of the gene to be transferred and have concerns about immunogenicity and safety. On the other hand, gene transfer using lipid particles is not limited in the gene to be transferred and can solve the above problems, so its development is actively underway.

[0004] As compounds to be contained in lipid particles, Patent Document 1 discloses compounds having an ester group, acetal group, or the like as a linking group connecting an aliphatic group and an amino group. Patent Document 2 also describes a compound having an alkylenediamine structure such as an ethylenediamine structure, and describes that lipid particles containing this compound exhibit a high nucleic acid encapsulation rate and excellent nucleic acid delivery.

[0005] International Publication No. 2010 / 054401 Pamphlet International Publication No. 2019 / 235635 Pamphlet

[0006] Further exploration is being conducted into lipid particles that can function as vectors and the compounds that make them up, and there is a desire to develop compounds that can achieve excellent nucleic acid delivery.

[0007] In view of the above circumstances, the present invention aims to provide a compound or a salt thereof that constitutes lipid particles that can achieve a high nucleic acid encapsulation rate and excellent nucleic acid delivery, and lipid particles and pharmaceutical compositions that use the compound and can achieve a high nucleic acid encapsulation rate and excellent nucleic acid delivery.

[0008] As a result of intensive research to solve the above problems, the present inventors have confirmed that lipid particles prepared using a compound represented by the following formula (1) or a salt thereof exhibit a high nucleic acid encapsulation rate and excellent nucleic acid delivery, and have completed the present invention. According to the present invention, the following inventions are provided:

[0009] <1> A compound represented by the following formula (1) or a salt thereof: In the formula, R １ and R ２ each independently represents a hydrocarbon group having 1 to 18 carbon atoms; R ３ represents a hydrocarbon group having 2 to 8 carbon atoms, and R １ , R ２ and R ３ The hydrocarbon group represented by is —OH, COOH, —NR ５１ R ５２ , -OC(O)OR ５３ , -C(O)O-R ５４ , —OC(O)—R ５５ , and -O-R ５６ and R ４ represents a hydrocarbon group having 1 to 8 carbon atoms, and R ５ and R ６ are each independently a hydrocarbon group having 1 to 8 carbon atoms, or -R ８ -L １ -R ９ where R ５ and R ６ and R are both hydrocarbon groups having 1 to 8 carbon atoms, ７ is -R １０ -L ２ -R １１ -L ３ -R １２ indicates, R ５１ and R ５２ each independently represents a hydrocarbon group having 1 to 8 carbon atoms; R５３ , R ５４ , R ５５ , and R ５６ each independently represents a hydrocarbon group having 1 to 24 carbon atoms; R ５３ , R ５４ , R ５５ , and R ５６ The hydrocarbon group represented by is an aryl group having 6 to 20 carbon atoms or -S-R ５８ The aryl group having 6 to 20 carbon atoms may be substituted with —OH, —COOH, —NR ５１ R ５２ , -OC(O)OR ５３ , -C(O)O-R ５４ , —OC(O)—R ５５ , -O-R ５６ or -(C1-C12 hydrocarbon group)-R ５７ and R ５８ represents a hydrocarbon group having 1 to 12 carbon atoms, and R ５７ is -OH, COOH, -NR ６１ R ６２ , -OC(O)OR ６３ , -C(O)O-R ６４ , —OC(O)—R ６５ , -O-R ６６ Indicates. ６１ and R ６２ each independently represents a hydrocarbon group having 1 to 8 carbon atoms; R ６３ , R ６４ , R ６５ , and R ６６ each independently represents a hydrocarbon group having 1 to 24 carbon atoms; R ６３ , R ６４ , R ６５ , and R ６６ The hydrocarbon group represented by is an aryl group having 6 to 20 carbon atoms or -S-R ６８ The aryl group having 6 to 20 carbon atoms may be substituted with —OH, —COOH, —NR ６１ R ６２ , -OC(O)OR ６３ , -C(O)O-R ６４ , —OC(O)—R ６５ , -O-R ６６ or -(C1-C12 hydrocarbon group)-R６７ and R ６８ represents a hydrocarbon group having 1 to 12 carbon atoms; L １ , L ２ , and L ３ R each independently represents —OC(O)O—, —C(O)O—, —OC(O)—, or —O—. ８ represents a hydrocarbon group having 1 to 12 carbon atoms, and R ９ represents a hydrocarbon group having 1 to 24 carbon atoms, R １０ represents a hydrocarbon group having 1 to 8 carbon atoms, and R １１ represents a hydrocarbon group having 1 to 24 carbon atoms, R １２ represents a hydrocarbon group having 1 to 24 carbon atoms, R ９ , and R １２ The hydrocarbon group represented by is an aryl group, —OC(O)O—R ５３ , -C(O)O-R ５４ , —OC(O)—R ５５ , or -S-R ５８ and R ５３ , R ５４ , R ５５ , and R ５８ is defined as above, and R １１ The hydrocarbon group represented by is —OC(O)O—R ５３ , -C(O)O-R ５４ or —OC(O)—R ５５ and R ５３ , R ５４ , and R ５５ The definition of is as above. <2> R １ and R ２ each independently represents a hydrocarbon group having 1 to 3 carbon atoms; R ３ represents a hydrocarbon group having 2 to 4 carbon atoms, and R １ , R ２ and R ３ The hydrocarbon group represented by R may be substituted with —OH, ４ represents a hydrocarbon group having 1 to 8 carbon atoms, and R ５ and R ６ are each independently a hydrocarbon group having 1 to 8 carbon atoms, or -R ８ -L １ -R ９where R ５ and R ６ and R are both hydrocarbon groups having 1 to 8 carbon atoms, ７ But, -R １０ -L ２ -R １１ -L ３ -R １２ indicates, L １ , and L ３ each independently represents —C(O)O— or —OC(O)—. ２ represents —OC(O)O—, —C(O)O—, or —OC(O)—. ８ represents a hydrocarbon group having 1 to 8 carbon atoms, and R ９ represents a hydrocarbon group having 1 to 16 carbon atoms, and R １０ represents a hydrocarbon group having 1 to 8 carbon atoms, and R １１ represents a hydrocarbon group having 1 to 9 carbon atoms, and R １２ represents a hydrocarbon group having 1 to 16 carbon atoms, and R ９ , and R １２ The hydrocarbon group represented by is an aryl group or -S-R ５８ and R ５８ represents a hydrocarbon group having 1 to 8 carbon atoms, and R １１ The hydrocarbon group represented by is —C(O)O—R ５５ , or -OC(O)-R ５６ and R ５５ , and R ５６ each independently represents a hydrocarbon group having 1 to 16 carbon atoms; R ５５ , and R ５６ The hydrocarbon group represented by is an aryl group having 6 to 20 carbon atoms or -S-R ５８ and R ５８ The definition of is as above. <3> The compound or salt thereof according to <1>, which is a compound represented by the following formula (1-1) or a salt thereof: In the formula, R １ and R ２ each independently represents a hydrocarbon group having 1 to 18 carbon atoms; R ３ represents a hydrocarbon group having 2 to 8 carbon atoms, and R １ , R２ and R ３ The hydrocarbon group represented by is —OH, COOH, —NR ５１ R ５２ , -OC(O)OR ５３ , -C(O)O-R ５４ , —OC(O)—R ５５ , or -O-R ５６ and R ４ represents a hydrocarbon group having 1 to 8 carbon atoms, and R ５ and R ６ are each independently a hydrocarbon group having 1 to 8 carbon atoms, or -R ８ -L １ -R ９ where R ５ and R ６ and L are both hydrocarbon groups having 1 to 8 carbon atoms, １ represents —OC(O)O—, —C(O)O—, —OC(O)—, or —O—, R ８ represents a hydrocarbon group having 1 to 12 carbon atoms, and R ９ represents a hydrocarbon group having 1 to 24 carbon atoms, and R ９ The hydrocarbon group represented by is an aryl group, —OC(O)O—R ５３ , -C(O)O-R ５４ , —OC(O)—R ５５ , or -S-R ５８ and R ５１ and R ５２ each independently represents a hydrocarbon group having 1 to 8 carbon atoms; R ５３ , R ５４ , R ５５ , and R ５６ each independently represents a hydrocarbon group having 1 to 24 carbon atoms; R ５３ , R ５４ , R ５５ , and R ５６ The hydrocarbon group represented by is an aryl group having 6 to 20 carbon atoms or -S-R ５８ The aryl group having 6 to 20 carbon atoms may be substituted with —OH, —COOH, —NR ５１ R ５２ , -OC(O)OR ５３ , -C(O)O-R ５４ , —OC(O)—R５５ , -O-R ５６ or -(C1-C12 hydrocarbon group)-R ５７ and R ５８ represents a hydrocarbon group having 1 to 12 carbon atoms, and R ５７ is -OH, COOH, -NR ５１ R ５２ , -OC(O)OR ５３ , -C(O)O-R ５４ , —OC(O)—R ５５ , -O-R ５６ Indicates. １３ represents a hydrocarbon group having 1 to 8 carbon atoms, and R １４ is -R １５ -L ５ -R １６ indicates R １５ represents a hydrocarbon group having 1 to 24 carbon atoms, and L ５ represents —OC(O)O—, —C(O)O—, —OC(O)—, or —O—, and R １６ represents a hydrocarbon group having 1 to 24 carbon atoms, R １５ The hydrocarbon group having 1 to 24 carbon atoms represented by is —OC(O)O—R ５３ , -C(O)O-R ５４ , or —OC(O)—R ５５ and R ５３ , R ５４ , and R ５５ is defined as above, and R １６ The hydrocarbon group having 1 to 24 carbon atoms represented by is an aryl group having 6 to 20 carbon atoms, ５３ , -C(O)O-R ５４ , —OC(O)—R ５５ , or -S-R ５８ and R ５３ , R ５４ , R ５５ , and R ５８ The definition of is as above. <4> In formula (1-1), R １ and R ２ each independently represents a hydrocarbon group having 1 to 3 carbon atoms; R ３ represents a hydrocarbon group having 2 to 4 carbon atoms, and R １ , R ２ and R３ The hydrocarbon group represented by R may be substituted with —OH, ４ represents a hydrocarbon group having 1 to 8 carbon atoms, and R ５ and R ６ are each independently a hydrocarbon group having 1 to 8 carbon atoms, or -R ８ -L １ -R ９ where R ５ and R ６ and L are both hydrocarbon groups having 1 to 8 carbon atoms, １ represents —C(O)O— or —OC(O)—, R ８ represents a hydrocarbon group having 1 to 8 carbon atoms, and R ９ represents a hydrocarbon group having 1 to 18 carbon atoms, and R ９ The hydrocarbon group represented by is an aryl group having 6 to 20 carbon atoms, or -S-R ５８ and R ５８ represents a hydrocarbon group having 1 to 8 carbon atoms, and R １３ represents a hydrocarbon group having 1 to 8 carbon atoms, and R １４ is -R １５ -L ５ -R １６ indicates R １５ represents a hydrocarbon group having 1 to 18 carbon atoms; L ５ represents -OC(O)O-, and R １６ represents a hydrocarbon group having 1 to 18 carbon atoms, and R １５ The hydrocarbon group having 1 to 18 carbon atoms represented by is —C(O)O—R ５５ , or —OC(O)—R ５６ and R ５５ , and R ５６ each independently represents a hydrocarbon group having 1 to 16 carbon atoms; R ５５ , and R ５６ The hydrocarbon group represented by is an aryl group having 6 to 20 carbon atoms or -S-R ５８ and R ５８ is defined as above, and R １６ The hydrocarbon group having 1 to 18 carbon atoms represented by is an aryl group or -S-R ５８ and R ５８The definition of is as above. <5> The compound or salt thereof according to <1>, which is a compound represented by the following formula (1-2) or a salt thereof: In the formula, R １ and R ２ each independently represents a hydrocarbon group having 1 to 18 carbon atoms; R ３ represents a hydrocarbon group having 2 to 8 carbon atoms, and R １ , R ２ and R ３ The hydrocarbon group represented by is —OH, COOH, —NR ５１ R ５２ , -OC(O)OR ５３ , -C(O)O-R ５４ , —OC(O)—R ５５ , or -O-R ５６ and R ４ and R ８ each independently represents a hydrocarbon having 1 to 8 carbon atoms; R ２１ and R ２２ each independently represents a hydrocarbon group having 1 to 18 carbon atoms; R ２３ and R ２４ each independently represents a hydrocarbon group having 1 to 12 carbon atoms; R ２５ and R ２６ each independently represents a hydrocarbon group having 1 to 24 carbon atoms; L ２１ and L ２２ each independently represents —OC(O)O—, —C(O)O—, —OC(O)—, or —O—; R ２５ and R ２６ The hydrocarbon group represented by is an aryl group having 6 to 20 carbon atoms, —OC(O)O—R ５３ , -C(O)O-R ５４ , —OC(O)—R ５５ , or -S-R ５８ and R ５１ and R ５２ each independently represents a hydrocarbon group having 1 to 8 carbon atoms; R ５３ , R ５４ , R ５５ and R ５６each independently represents a hydrocarbon group having 1 to 18 carbon atoms, and the aryl group having 6 to 20 carbon atoms is OH, COOH, —NR ５１ R ５２ , -OC(O)OR ５３ , -C(O)O-R ５４ , —OC(O)—R ５５ , -O-R ５６ or -(C1-C12 hydrocarbon group)-R ５７ and R ５７ is -OH, COOH, -NR ５１ R ５２ , -OC(O)OR ５３ , -C(O)O-R ５４ , —OC(O)—R ５５ , -O-R ５６ Indicates. ５８ represents a hydrocarbon group having 1 to 12 carbon atoms. １ and R ２ each independently represents a hydrocarbon group having 1 to 3 carbon atoms; R １ and R ２ The hydrocarbon group represented by R may be substituted with —OH, ３ represents a hydrocarbon group having 2 to 4 carbon atoms, and R ４ and R ８ each independently represents a hydrocarbon having 1 to 8 carbon atoms; R ２１ and R ２２ each independently represents a hydrocarbon group having 1 to 8 carbon atoms; R ２３ and R ２４ each independently represents a hydrocarbon group having 1 to 8 carbon atoms; R ２５ and R ２６ each independently represents a hydrocarbon group having 1 to 16 carbon atoms; ２１ and L ２２ <7> The compound or salt thereof according to <5>, wherein, in formula (1-2), R １ and R ２ each independently represents a hydrocarbon group having 1 to 3 carbon atoms; R ３ represents a hydrocarbon group having 2 to 4 carbon atoms, and R ４ and R８ each independently represents a hydrocarbon having 1 to 8 carbon atoms; R ２１ and R ２２ each independently represents a hydrocarbon group having 1 to 6 carbon atoms; R ２３ and R ２４ each independently represents a hydrocarbon group having 1 to 8 carbon atoms; R ２５ and R ２６ each independently represents a hydrocarbon group having 1 to 12 carbon atoms; ２１ and L ２２ each independently represent —C(O)O— or —OC(O)—. <8> The compound or salt thereof according to <1>, which is a compound represented by the following formula (1-3) or a salt thereof: In the formula, R １ and R ２ each independently represents a hydrocarbon group having 1 to 18 carbon atoms; R ３ represents a hydrocarbon group having 2 to 8 carbon atoms, and R １ , R ２ and R ３ The hydrocarbon group represented by is —OH, COOH, —NR ５１ R ５２ , -OC(O)OR ５３ , -C(O)O-R ５４ , —OC(O)—R ５５ , or -O-R ５６ and R ４ and R ８ each independently represents a hydrocarbon group having 1 to 8 carbon atoms; R ３１ , R ３２ , R ３３ , and R ３４ each independently represents a hydrocarbon group having 1 to 12 carbon atoms; R ３５ , R ３６ , R ３７ , and R ３８ each independently represents a hydrocarbon group having 1 to 24 carbon atoms; L ３１ , L ３２ , L ３３ , and L ３４ each independently represents —OC(O)O—, —C(O)O—, —OC(O)—, or —O—; R ３５ , R３６ , R ３７ , and R ３８ The hydrocarbon group represented by is an aryl group having 6 to 20 carbon atoms, —OC(O)O—R ５３ , -C(O)O-R ５４ , —OC(O)—R ５５ , or S-R ５８ and R ５１ and R ５２ each independently represents a hydrocarbon group having 1 to 8 carbon atoms; R ５３ , R ５４ , R ５５ , and R ５６ each independently represents a hydrocarbon group having 1 to 18 carbon atoms, and the aryl group having 6 to 20 carbon atoms is OH, COOH, —NR ５１ R ５２ , -OC(O)OR ５３ , -C(O)O-R ５４ , —OC(O)—R ５５ , -O-R ５６ or -(C1-C12 hydrocarbon group)-R ５７ and R ５７ is -OH, COOH, -NR ５１ R ５２ , -OC(O)OR ５３ , -C(O)O-R ５４ , —OC(O)—R ５５ , -O-R ５６ Indicates. ５８ represents a hydrocarbon group having 1 to 12 carbon atoms. <9> In formula (1-3), R １ and R ２ each independently represents a hydrocarbon group having 1 to 3 carbon atoms; R １ and R ２ The hydrocarbon group represented by R may be substituted with —OH, ３ represents a hydrocarbon group having 2 to 4 carbon atoms, and R ４ and R ８ each independently represents a hydrocarbon group having 1 to 8 carbon atoms; R ３１ , R ３２ , R ３３ , and R ３４ each independently represents a hydrocarbon group having 1 to 8 carbon atoms; R３５ , R ３６ , R ３７ , and R ３８ each independently represents a hydrocarbon group having 1 to 16 carbon atoms; ３１ , L ３２ , L ３３ , and L ３４ each independently represents —C(O)O— or —OC(O)—, R ３５ , R ３６ , R ３７ , and R ３８ The hydrocarbon group represented by is an aryl group having 6 to 20 carbon atoms, or S—R ５８ and R ５８ <10> The compound or salt thereof according to <8>, wherein, in formula (1-3), R １ and R ２ each independently represents a hydrocarbon group having 1 to 3 carbon atoms; R ３ represents a hydrocarbon group having 2 to 4 carbon atoms, and R ４ and R ８ each independently represents a hydrocarbon group having 1 to 8 carbon atoms; R ３１ , R ３２ , R ３３ , and R ３４ each independently represents a hydrocarbon group having 1 to 3 carbon atoms; R ３５ , R ３６ , R ３７ , and R ３８ each independently represents a hydrocarbon group having 1 to 12 carbon atoms; ３１ , L ３２ , L ３３ , and L ３４ each independently represents —C(O)O— or —OC(O)—, R ３５ , R ３６ , R ３７ , and R ３８ The hydrocarbon group represented by is —S—R ５８ and R ５８ <11> The compound or salt thereof according to <8>, wherein, in formula (1-3), R １ and R ２each independently represents a hydrocarbon group having 1 to 3 carbon atoms; R ３ represents a hydrocarbon group having 2 to 4 carbon atoms, and R ４ and R ８ each independently represents a hydrocarbon group having 1 to 8 carbon atoms; R ３１ , R ３２ , R ３３ , and R ３４ each independently represents a hydrocarbon group having 1 to 3 carbon atoms; R ３５ , R ３６ , R ３７ , and R ３８ are each independently -S-R ５８ represents a hydrocarbon group having 1 to 12 carbon atoms substituted with R ５８ represents a hydrocarbon group having 1 to 8 carbon atoms; L ３１ , L ３２ , L ３３ , and L ３４ <12> The compound or salt thereof according to <8>, wherein, in formula (1-3), R １ and R ２ each independently represents a hydrocarbon group having 1 to 3 carbon atoms; R ３ represents a hydrocarbon group having 2 to 4 carbon atoms, and R ４ and R ８ each independently represents a hydrocarbon group having 1 to 8 carbon atoms; R ３１ , R ３２ , R ３３ , and R ３４ each independently represents a hydrocarbon group having 1 to 3 carbon atoms; R ３５ , R ３６ , R ３７ , and R ３８ each independently represents a hydrocarbon group having 1 to 12 carbon atoms; ３１ , L ３２ , L ３３ , and L ３４each independently represent —C(O)O— or —OC(O)—. <13> The compound or a salt thereof according to <8>, <14> The compound or a salt thereof according to <13 ... Bis(2-butyloctyl) 11-(2-(diethylamino)ethyl)-5,17-dihexyl-7,15-dioxo-6,8,14,16-tetraoxa-11-azahenicosanedioate; Bis(2-pentylheptyl)11-(2-(diethylamino)ethyl)-5,17-dihexyl-7,15-dioxo-6,8,14,16-tetraoxa-11-azahenicosanedioate; Bis(2-pentylheptyl)11-(2-(dimethylamino)ethyl)-5,17-dihexyl-7,15-dioxo-6,8,14,16-tetraoxa-11-azahenicosanedioate; Bis(2-pentylheptyl)11-(3-(diethylamino)propyl)-5,17-dihexyl-7,15-dioxo-6,8,14,16-tetraoxa-11-azahenicosanedioate; Bis(2-pentylheptyl)11-(4-(diethylamino)butyl)-5,17-dihexyl-7,15-dioxo-6,8,14,16-tetraoxa-11-azahenicosanedioate; Bis(2-pentylheptyl)12-(2-(diethylamino)ethyl)-5,19-dihexyl-7,17-dioxo-6,8,16,18-tetraoxa-12-azatricosane dioate; Bis(2-pentylheptyl)13-(2-(diethylamino)ethyl)-5,21-dihexyl-7,19-dioxo-6,8,18,20-tetraoxa-13-azapentacosane dioate; Bis(2-pentylheptyl)10-(2-(diethylamino)ethyl)-4,16-dihexyl-6,14-dioxo-5,7,13,15-tetraoxa-10-azanonadecanedioate; Bis(2-pentylheptyl)11-(2-(diethylamino)ethyl)-5,17-dimethyl-7,15-dioxo-6,8,14,16-tetraoxa-11-azahenicosanedioate; Bis(2-pentylheptyl)11-(2-(diethylamino)ethyl)-5,17-diethyl-7,15-dioxo-6,8,14,16-tetraoxa-11-azahenicosanedioate; Bis(2-pentylheptyl)11-(2-(diethylamino)ethyl)-7,15-dioxo-5,17-dipropyl-6,8,14,16-tetraoxa-11-azahenicosanedioate; Bis(2-pentylheptyl)5,17-dibutyl-11-(2-(diethylamino)ethyl)-7,15-dioxo-6,8,14,16-tetraoxa-11-azahenicosanedioate; Bis(2-pentylheptyl)11-(2-(diethylamino)ethyl)-7,15-dioxo-5,17-dipentyl-6,8,14,16-tetraoxa-11-azahenicosanedioate; Bis(2-pentylheptyl)11-(2-(diethylamino)ethyl)-5,17-diheptyl-7,15-dioxo-6,8,14,16-tetraoxa-11-azahenicosanedioate; Bis(2-((3r,5r,7r)-adamantan-1-yl)ethyl)11-(2-(diethylamino)ethyl)-5,17-dihexyl-7,15-dioxo-6,8,14,16-tetraoxa-11-azahenicosanedioate; Bis(2-pentylheptyl)11-(3-(diethylamino)propyl)-7,15-dioxo-5,17-dipropyl-6,8,14,16-tetraoxa-11-azahenicosanedioate; Diheptyl 11-(2-(diethylamino)ethyl)-5,17-bis(4-(heptyloxy)-4-oxobutyl)-7,15-dioxo-6,8,14,16-tetraoxa-11-azahenicosanedioate; Dihexyl 11-(2-(diethylamino)ethyl)-5,17-bis(4-(hexyloxy)-4-oxobutyl)-7,15-dioxo-6,8,14,16-tetraoxa-11-azahenicosanedioate; Dioctyl 11-(2-(diethylamino)ethyl)-5,17-bis(4-(octyloxy)-4-oxobutyl)-7,15-dioxo-6,8,14,16-tetraoxa-11-azahenicosanedioate; Dinonyl 11-(2-(diethylamino)ethyl)-5,17-bis(4-(nonyloxy)-4-oxobutyl)-7,15-dioxo-6,8,14,16-tetraoxa-11-azahenicosanedioate; 1-heptyl 21-hexyl 11-(2-(diethylamino)ethyl)-5-(4-(heptyloxy)-4-oxobutyl)-17-(4-(hexyloxy)-4-oxobutyl)-7,15-dioxo-6,8,14,16-tetraoxa-11-azahenicosanedioate; Diheptyl 11-(3-(diethylamino)propyl)-5,17-bis(4-(heptyloxy)-4-oxobutyl)-7,15-dioxo-6,8,14,16-tetraoxa-11-azahenicosanedioate; Diheptyl 10-(2-(diethylamino)ethyl)-4,16-bis(3-(heptyloxy)-3-oxopropyl)-6,14-dioxo-5,7,13,15-tetraoxa-10-azanonadecanedioate; 1-Hexyl 21-octyl 11-(2-(diethylamino)ethyl)-5-(4-(hexyloxy)-4-oxobutyl)-17-(4-(octyloxy)-4-oxobutyl)-7,15-dioxo-6,8,14,16-tetraoxa-11-azahenicosanedioate; Bis(2-(hexylthio)ethyl)11-(2-(diethylamino)ethyl)-5,17-bis(4-(2-(hexylthio)ethoxy)-4-oxobutyl)-7,15-dioxo-6,8,14,16-tetraoxa-11-azahenicosanedioate; Bis(8-(methylthio)octyl)11-(2-(diethylamino)ethyl)-5,17-bis(4-((8-(methylthio)octyl)oxy)-4-oxobutyl)-7,15-dioxo-6,8,14,16-tetraoxa-11-azahenicosanedioate; <14> A lipid particle comprising the compound or salt thereof according to any one of <1> to <13> and a lipid. <15> The lipid particle according to <14>, wherein the lipid is at least one type of lipid selected from the group consisting of sterols and lipids having a nonionic hydrophilic polymer chain. <16> The lipid particle according to <14> or <15>, further comprising a neutral lipid. <17> The lipid particle according to any one of <14> to <16>, further comprising a nucleic acid. <18> The lipid particle according to <17>, wherein the nucleic acid comprises a nucleic acid having 50 or more bases. <19> A pharmaceutical composition comprising the lipid particle according to any one of <14> to <18> as an active ingredient.

[0010] By using the compound of the present invention, it is possible to produce lipid particles and pharmaceutical compositions that can achieve a high nucleic acid encapsulation rate and excellent nucleic acid delivery. The lipid particles and pharmaceutical compositions of the present invention can achieve a high nucleic acid encapsulation rate and excellent nucleic acid delivery.

[0011] The present invention will be described in detail below. In this specification, the symbol "to" indicates a range that includes the numerical values ​​before and after it as the minimum and maximum values, respectively.

[0012] <Compound of the Present Invention> The compound of the present invention is represented by the following formula (1).

[0013] In the formula, R １ and R ２ each independently represents a hydrocarbon group having 1 to 18 carbon atoms; R ３ represents a hydrocarbon group having 2 to 8 carbon atoms, and R １ , R ２ and R ３ The hydrocarbon group represented by is —OH, COOH, —NR ５１ R ５２ , -OC(O)OR ５３ , -C(O)O-R ５４ , —OC(O)—R５５ , and -O-R ５６ and R ４ represents a hydrocarbon group having 1 to 8 carbon atoms, and R ５ and R ６ are each independently a hydrocarbon group having 1 to 8 carbon atoms, or -R ８ -L １ -R ９ where R ５ and R ６ and R are both hydrocarbon groups having 1 to 8 carbon atoms, ７ is -R １０ -L ２ -R １１ -L ３ -R １２ indicates, R ５１ and R ５２ each independently represents a hydrocarbon group having 1 to 8 carbon atoms; R ５３ , R ５４ , R ５５ , and R ５６ each independently represents a hydrocarbon group having 1 to 24 carbon atoms; R ５３ , R ５４ , R ５５ , and R ５６ The hydrocarbon group represented by is an aryl group having 6 to 20 carbon atoms or -S-R ５８ The aryl group having 6 to 20 carbon atoms may be substituted with —OH, —COOH, —NR ５１ R ５２ , -OC(O)OR ５３ , -C(O)O-R ５４ , —OC(O)—R ５５ , -O-R ５６ or -(C1-C12 hydrocarbon group)-R ５７ and R ５８ represents a hydrocarbon group having 1 to 12 carbon atoms, and R ５７ is -OH, COOH, -NR ６１ R ６２ , -OC(O)OR ６３ , -C(O)O-R ６４ , —OC(O)—R ６５ , -O-R ６６ Indicates. ６１and R ６２ each independently represents a hydrocarbon group having 1 to 8 carbon atoms; R ６３ , R ６４ , R ６５ , and R ６６ each independently represents a hydrocarbon group having 1 to 24 carbon atoms; R ６３ , R ６４ , R ６５ , and R ６６ The hydrocarbon group represented by is an aryl group having 6 to 20 carbon atoms or -S-R ６８ The aryl group having 6 to 20 carbon atoms may be substituted with —OH, —COOH, —NR ６１ R ６２ , -OC(O)OR ６３ , -C(O)O-R ６４ , —OC(O)—R ６５ , -O-R ６６ or -(C1-C12 hydrocarbon group)-R ６７ and R ６８ represents a hydrocarbon group having 1 to 12 carbon atoms; L １ , L ２ , and L ３ R each independently represents —OC(O)O—, —C(O)O—, —OC(O)—, or —O—. ８ represents a hydrocarbon group having 1 to 12 carbon atoms, and R ９ represents a hydrocarbon group having 1 to 24 carbon atoms, R １０ represents a hydrocarbon group having 1 to 8 carbon atoms, and R １１ represents a hydrocarbon group having 1 to 24 carbon atoms, R １２ represents a hydrocarbon group having 1 to 24 carbon atoms, R ９ , and R １２ The hydrocarbon group represented by is an aryl group, —OC(O)O—R ５３ , -C(O)O-R ５４ , —OC(O)—R ５５ , or -S-R ５８ and R ５３ , R ５４ , R ５５ , and R ５８ is defined as above, and R １１ The hydrocarbon group represented by is —OC(O)O—R ５３, -C(O)O-R ５４ or —OC(O)—R ５５ and R ５３ , R ５４ , and R ５５ The definition of is as above.

[0014] The hydrocarbon group having 1 to 24 carbon atoms, the hydrocarbon group having 1 to 18 carbon atoms, the hydrocarbon group having 1 to 12 carbon atoms, the hydrocarbon group having 2 to 8 carbon atoms, and the hydrocarbon group having 1 to 8 carbon atoms are preferably an alkyl group, an alkenyl group, or an alkynyl group, respectively.

[0015] The alkyl group may be linear or branched, and may be linear or cyclic. Specific examples of the alkyl group include a methyl group, an ethyl group, a propyl group, an isopropyl group, a cyclopropyl group, a butyl group, an isobutyl group, a tert-butyl group, a cyclobutyl group, a pentyl group, a cyclopentyl group, a hexyl group, a cyclohexyl group, a heptyl group, an octyl group, a nonyl group, a decyl group, an undecyl group, a dodecyl group, a tridecyl group, a trimethyldodecyl group (preferably a 3,7,11-trimethyldodecyl group), a tetradecyl group, a pentadecyl group, a hexadecyl group, a tetramethylhexadecyl group (preferably a 3,7,11,15-tetramethylhexadecyl group), a heptadecyl group, an octadecyl group, a 2-butylhexyl group, and a 2-butyloctyl group. , 1-pentylhexyl group, 2-pentylheptyl group, 3-pentyloctyl group, 1-hexylheptyl group, 1-hexylnonyl group, 2-hexyloctyl group, 2-hexyldecyl group, 3-hexylnonyl group, 1-heptyloctyl group, 2-heptylnonyl group, 2-heptylundecyl group, 3-heptyldecyl group, 1-octylnonyl group, 2-octyldecyl group, 2-octyldodecyl group, 3-octylundecyl group, 2-nonylundecyl group, 3-nonyldodecyl group, 2-decyldodecyl group, 2-decyltetradecyl group, 3-decyltridecyl group, 2-(4,4-dimethylpentan-2-yl)-5,7,7-trimethyloctyl group, and the like.

[0016] The alkenyl group may be linear or branched, linear or cyclic. Specific examples include allyl, prenyl, pentenyl, hexenyl, heptenyl, octenyl, nonenyl (preferably, (Z)-2-nonenyl or (E)-2-nonenyl), decenyl, undecenyl, dodecenyl, dodecadienyl, tridecenyl (preferably, (Z)-tridec-8-enyl), tetradecenyl (preferably, tetradec-9-enyl), and pentadecenyl (preferably, (Z)-pentadecen-8-enyl). , a hexadecenyl group (preferably a (Z)-hexadec-9-enyl group), a hexadecadienyl group, a heptadecenyl group (preferably a (Z)-heptadeca-8-enyl group), a heptadecadienyl group (preferably a (8Z,11Z)-heptadeca-8,11-dienyl group), an octadecenyl group (preferably a (Z)-octadec-9-enyl group), an octadecadienyl group (preferably a (9Z,12Z)-octadeca-9,12-dienyl group), and the like.

[0017] The alkynyl group may be linear or branched, open-chain or cyclic, and specific examples thereof include a propargyl group, a butynyl group, a pentynyl group, a hexynyl group, a heptynyl group, an octynyl group, a nonynyl group, a decynyl group, an undecynyl group, a dodecynyl group, a tetradecynyl group, a pentadecynyl group, a hexadecynyl group, a heptadecynyl group, and an octadecynyl group.

[0018] Preferably, all of the above alkenyl groups have one or two double bonds, and preferably, all of the alkynyl groups have one or two triple bonds.

[0019] -(C1 to C12 hydrocarbon group)-R ６７The hydrocarbon group having 1 to 12 carbon atoms in the formula (I) is preferably an alkylene group having 1 to 12 carbon atoms or an alkenylene group having 2 to 12 carbon atoms. The alkylene group having 1 to 12 carbon atoms and the alkenylene group having 2 to 12 carbon atoms may be linear or branched, and may be linear or cyclic. Specific examples include a methylene group, an ethylene group, a trimethylene group, a tetramethylene group, a pentamethylene group, a hexamethylene group, a heptamethylene group, an octamethylene group, a nonamethylene group, a decamethylene group, and an undecamethylene group.

[0020] The aryl group preferably has 6 to 20 carbon atoms, more preferably 6 to 18 carbon atoms, and even more preferably 6 to 10 carbon atoms. Specific examples include a phenyl group, a naphthyl group, an anthracenyl group, and a phenanthrenyl group.

[0021] R １ and R ２ R each independently represents a hydrocarbon group having preferably 1 to 12 carbon atoms, more preferably a hydrocarbon group having 1 to 6 carbon atoms, and even more preferably a hydrocarbon group having 1 to 3 carbon atoms. ３ R preferably represents a hydrocarbon group having 2 to 6 carbon atoms, and more preferably represents a hydrocarbon group having 2 to 4 carbon atoms. １ , R ２ and R ３ The hydrocarbon group represented by may preferably be substituted with —OH.

[0022] L １ , and L ３ each independently preferably represents —C(O)O— or —OC(O)—. ２ preferably represents —OC(O)O—, —C(O)O—, or —OC(O)—.

[0023] R ８ R preferably represents a hydrocarbon group having 1 to 10 carbon atoms, and more preferably represents a hydrocarbon group having 1 to 8 carbon atoms. ９ R preferably represents a hydrocarbon group having 1 to 20 carbon atoms, and more preferably represents a hydrocarbon group having 1 to 16 carbon atoms. １１ R preferably represents a hydrocarbon group having 1 to 16 carbon atoms, and more preferably represents a hydrocarbon group having 1 to 9 carbon atoms.１２ R preferably represents a hydrocarbon group having 1 to 20 carbon atoms, and more preferably represents a hydrocarbon group having 1 to 16 carbon atoms. ９ , and R １２ The hydrocarbon group represented by is preferably an aryl group or —S—R ５８ where R ５８ R preferably represents a hydrocarbon group having 1 to 8 carbon atoms. １１ The hydrocarbon group represented by is preferably —C(O)O—R ５５ , or -OC(O)-R ５６ where R ５５ , and R ５６ R each independently represents a hydrocarbon group having 1 to 16 carbon atoms; ５５ , and R ５６ The hydrocarbon group represented by is preferably an aryl group having 6 to 20 carbon atoms or —S—R ５８ and R ５８ The definition of is as above.

[0024] The compound represented by formula (1) is preferably, as a first example, a compound represented by the following formula (1-1): In the formula, R １ and R ２ each independently represents a hydrocarbon group having 1 to 18 carbon atoms; R ３ represents a hydrocarbon group having 2 to 8 carbon atoms, and R １ , R ２ and R ３ The hydrocarbon group represented by is —OH, COOH, —NR ５１ R ５２ , -OC(O)OR ５３ , -C(O)O-R ５４ , —OC(O)—R ５５ , or -O-R ５６ and R ４ represents a hydrocarbon group having 1 to 8 carbon atoms, and R ５ and R ６ are each independently a hydrocarbon group having 1 to 8 carbon atoms, or -R ８ -L １ -R ９ where R ５ and R６ and L are both hydrocarbon groups having 1 to 8 carbon atoms, １ represents —OC(O)O—, —C(O)O—, —OC(O)—, or —O—, R ８ represents a hydrocarbon group having 1 to 12 carbon atoms, and R ９ represents a hydrocarbon group having 1 to 24 carbon atoms, and R ９ The hydrocarbon group represented by is an aryl group, —OC(O)O—R ５３ , -C(O)O-R ５４ , —OC(O)—R ５５ , or -S-R ５８ and R ５１ and R ５２ each independently represents a hydrocarbon group having 1 to 8 carbon atoms; R ５３ , R ５４ , R ５５ , and R ５６ each independently represents a hydrocarbon group having 1 to 24 carbon atoms; R ５３ , R ５４ , R ５５ , and R ５６ The hydrocarbon group represented by is an aryl group having 6 to 20 carbon atoms or -S-R ５８ The aryl group having 6 to 20 carbon atoms may be substituted with —OH, —COOH, —NR ５１ R ５２ , -OC(O)OR ５３ , -C(O)O-R ５４ , —OC(O)—R ５５ , -O-R ５６ or -(C1-C12 hydrocarbon group)-R ５７ and R ５８ represents a hydrocarbon group having 1 to 12 carbon atoms, and R ５７ is -OH, COOH, -NR ５１ R ５２ , -OC(O)OR ５３ , -C(O)O-R ５４ , —OC(O)—R ５５ , -O-R ５６ Indicates. １３ represents a hydrocarbon group having 1 to 8 carbon atoms, and R １４ is -R １５ -L ５ -R１６ indicates R １５ represents a hydrocarbon group having 1 to 24 carbon atoms, and L ５ represents —OC(O)O—, —C(O)O—, —OC(O)—, or —O—, and R １６ represents a hydrocarbon group having 1 to 24 carbon atoms, R １５ The hydrocarbon group having 1 to 24 carbon atoms represented by is —OC(O)O—R ５３ , -C(O)O-R ５４ , or —OC(O)—R ５５ and R ５３ , R ５４ , and R ５５ is defined as above, and R １６ The hydrocarbon group having 1 to 24 carbon atoms represented by is an aryl group having 6 to 20 carbon atoms, ５３ , -C(O)O-R ５４ , —OC(O)—R ５５ , or -S-R ５８ and R ５３ , R ５４ , R ５５ , and R ５８ The definition of is as above.

[0025] In formula (1-1), R １ and R ２ R each independently represents a hydrocarbon group having preferably 1 to 12 carbon atoms, more preferably a hydrocarbon group having 1 to 6 carbon atoms, and even more preferably a hydrocarbon group having 1 to 3 carbon atoms. ３ R preferably represents a hydrocarbon group having 2 to 6 carbon atoms, and more preferably represents a hydrocarbon group having 2 to 4 carbon atoms. １ , R ２ and R ３ The hydrocarbon group represented by may preferably be substituted with —OH.

[0026] L １ preferably represents —C(O)O— or —OC(O)—. ８ R preferably represents a hydrocarbon group having 1 to 10 carbon atoms, and more preferably represents a hydrocarbon group having 1 to 8 carbon atoms. ９ preferably represents a hydrocarbon group having 1 to 18 carbon atoms, and R９ The hydrocarbon group represented by is an aryl group having 6 to 20 carbon atoms, or -S-R ５８ may be substituted with R １４ is preferably —R １５ -L ５ -R １６ indicates R １５ represents a hydrocarbon group having 1 to 18 carbon atoms; L ５ represents -OC(O)O-, and R １６ represents a hydrocarbon group having 1 to 18 carbon atoms. １５ The hydrocarbon group having 1 to 18 carbon atoms represented by is preferably —C(O)O—R ５５ , or —OC(O)—R ５６ may be substituted with R ５５ , and R ５６ each independently represents a hydrocarbon group having 1 to 16 carbon atoms; R ５５ , and R ５６ The hydrocarbon group represented by is an aryl group having 6 to 20 carbon atoms or -S-R ５８ and R ５８ The definition of R is as above. １６ The hydrocarbon group having 1 to 18 carbon atoms represented by is preferably an aryl group or —S—R ５８ and R ５８ The definition of is as above.

[0027] A second example of the compound represented by formula (1) is preferably a compound represented by the following formula (1-2): In the formula, R １ and R ２ each independently represents a hydrocarbon group having 1 to 18 carbon atoms; R ３ represents a hydrocarbon group having 2 to 8 carbon atoms, and R １ , R ２ and R ３ The hydrocarbon group represented by is —OH, COOH, —NR ５１ R ５２ , -OC(O)OR ５３ , -C(O)O-R ５４ , —OC(O)—R ５５ , or -O-R ５６ and R ４ and R８ each independently represents a hydrocarbon having 1 to 8 carbon atoms; R ２１ and R ２２ each independently represents a hydrocarbon group having 1 to 18 carbon atoms; R ２３ and R ２４ each independently represents a hydrocarbon group having 1 to 12 carbon atoms; R ２５ and R ２６ each independently represents a hydrocarbon group having 1 to 24 carbon atoms; L ２１ and L ２２ each independently represents —OC(O)O—, —C(O)O—, —OC(O)—, or —O—; R ２５ and R ２６ The hydrocarbon group represented by is an aryl group having 6 to 20 carbon atoms, —OC(O)O—R ５３ , -C(O)O-R ５４ , —OC(O)—R ５５ , or -S-R ５８ and R ５１ and R ５２ each independently represents a hydrocarbon group having 1 to 8 carbon atoms; R ５３ , R ５４ , R ５５ and R ５６ each independently represents a hydrocarbon group having 1 to 18 carbon atoms, and the aryl group having 6 to 20 carbon atoms is OH, COOH, —NR ５１ R ５２ , -OC(O)OR ５３ , -C(O)O-R ５４ , —OC(O)—R ５５ , -O-R ５６ or -(C1-C12 hydrocarbon group)-R ５７ and R ５７ is -OH, COOH, -NR ５１ R ５２ , -OC(O)OR ５３ , -C(O)O-R ５４ , —OC(O)—R ５５ , -O-R ５６ Indicates. ５８ represents a hydrocarbon group having 1 to 12 carbon atoms.

[0028] In formula (1-2), R１ and R ２ R each independently represents a hydrocarbon group having preferably 1 to 12 carbon atoms, more preferably a hydrocarbon group having 1 to 6 carbon atoms, and even more preferably a hydrocarbon group having 1 to 3 carbon atoms. １ and R ２ The hydrocarbon group represented by may be substituted with -OH, but is more preferably a hydrocarbon group without any substituents.

[0029] R ３ represents a hydrocarbon group preferably having 2 to 6 carbon atoms, and more preferably a hydrocarbon group having 2 to 4 carbon atoms.

[0030] R ２１ and R ２２ R each independently represents a hydrocarbon group having preferably 1 to 12 carbon atoms, more preferably a hydrocarbon group having 1 to 8 carbon atoms, and even more preferably a hydrocarbon group having 1 to 6 carbon atoms. ２３ and R ２４ R each independently represents a hydrocarbon group preferably having 1 to 10 carbon atoms, more preferably a hydrocarbon group having 1 to 8 carbon atoms. ２５ and R ２６ each independently represents a hydrocarbon group having preferably 1 to 20 carbon atoms, more preferably a hydrocarbon group having 1 to 16 carbon atoms, and even more preferably a hydrocarbon group having 1 to 12 carbon atoms. ２１ and L ２２ are each independently preferably —C(O)O— or —OC(O)—.

[0031] A third example of the compound represented by formula (1) is preferably a compound represented by the following formula (1-3). In the formula, R １ and R ２ each independently represents a hydrocarbon group having 1 to 18 carbon atoms; R ３ represents a hydrocarbon group having 2 to 8 carbon atoms, and R １ , R ２ and R ３ The hydrocarbon group represented by is —OH, COOH, —NR ５１ R ５２ , -OC(O)OR ５３ , -C(O)O-R５４ , —OC(O)—R ５５ , or -O-R ５６ and R ４ and R ８ each independently represents a hydrocarbon group having 1 to 8 carbon atoms; R ３１ , R ３２ , R ３３ , and R ３４ each independently represents a hydrocarbon group having 1 to 12 carbon atoms; R ３５ , R ３６ , R ３７ , and R ３８ each independently represents a hydrocarbon group having 1 to 24 carbon atoms; L ３１ , L ３２ , L ３３ , and L ３４ each independently represents —OC(O)O—, —C(O)O—, —OC(O)—, or —O—; R ３５ , R ３６ , R ３７ , and R ３８ The hydrocarbon group represented by is an aryl group having 6 to 20 carbon atoms, —OC(O)O—R ５３ , -C(O)O-R ５４ , —OC(O)—R ５５ , or S-R ５８ and R ５１ and R ５２ each independently represents a hydrocarbon group having 1 to 8 carbon atoms; R ５３ , R ５４ , R ５５ , and R ５６ each independently represents a hydrocarbon group having 1 to 18 carbon atoms, and the aryl group having 6 to 20 carbon atoms is OH, COOH, —NR ５１ R ５２ , -OC(O)OR ５３ , -C(O)O-R ５４ , —OC(O)—R ５５ , -O-R ５６ or -(C1-C12 hydrocarbon group)-R ５７ and R ５７ is -OH, COOH, -NR ５１ R ５２ , -OC(O)OR５３ , -C(O)O-R ５４ , —OC(O)—R ５５ , -O-R ５６ Indicates. ５８ represents a hydrocarbon group having 1 to 12 carbon atoms.

[0032] In formula (1-3), R １ and R ２ R each independently represents a hydrocarbon group having preferably 1 to 12 carbon atoms, more preferably a hydrocarbon group having 1 to 6 carbon atoms, and even more preferably a hydrocarbon group having 1 to 3 carbon atoms. １ and R ２ The hydrocarbon group represented by may be substituted with -OH, but is more preferably a hydrocarbon group without any substituents.

[0033] R ３ represents a hydrocarbon group preferably having 2 to 6 carbon atoms, and more preferably a hydrocarbon group having 2 to 4 carbon atoms.

[0034] R ３１ , R ３２ , R ３３ , and R ３４ are each independently preferably a hydrocarbon group having 1 to 10 carbon atoms, more preferably a hydrocarbon group having 1 to 8 carbon atoms, and even more preferably a hydrocarbon group having 1 to 3 carbon atoms.

[0035] R ３５ , R ３６ , R ３７ , and R ３８ R each independently represents a hydrocarbon group having preferably 1 to 20 carbon atoms, more preferably a hydrocarbon group having 1 to 16 carbon atoms, and even more preferably a hydrocarbon group having 1 to 12 carbon atoms. ３５ , R ３６ , R ３７ , and R ３８ The hydrocarbon group represented by is preferably an aryl group having 6 to 20 carbon atoms, or S—R ５８ More preferably, it is substituted with -S-R ５８ may be substituted with R ３５ , R ３６ , R ３７ , and R ３８are each independently particularly preferably —S—R ５８ or a hydrocarbon group having 1 to 12 carbon atoms substituted with

[0036] L ３１ , L ３２ , L ３３ , and L ３４ are each independently preferably —C(O)O— or —OC(O)—.

[0037] R ５８ represents a hydrocarbon group preferably having 1 to 10 carbon atoms, and more preferably a hydrocarbon group having 1 to 8 carbon atoms.

[0038] The compound of the present invention may form a salt. Examples of salts of basic groups include salts with mineral acids such as hydrochloric acid, hydrobromic acid, nitric acid, and sulfuric acid; salts with organic carboxylic acids such as formic acid, acetic acid, citric acid, oxalic acid, fumaric acid, maleic acid, succinic acid, malic acid, tartaric acid, aspartic acid, trichloroacetic acid, and trifluoroacetic acid; and salts with sulfonic acids such as methanesulfonic acid, benzenesulfonic acid, p-toluenesulfonic acid, mesitylenesulfonic acid, and naphthalenesulfonic acid. Salts of acidic groups include, for example, salts with alkali metals such as sodium and potassium, salts with alkaline earth metals such as calcium and magnesium, ammonium salts, and salts with nitrogen-containing organic bases such as trimethylamine, triethylamine, tributylamine, pyridine, N,N-dimethylaniline, N-methylpiperidine, N-methylmorpholine, diethylamine, dicyclohexylamine, procaine, dibenzylamine, N-benzyl-β-phenethylamine, 1-ephenamine, and N,N'-dibenzylethylenediamine. Of the above-mentioned salts, preferred salts include pharmacologically acceptable salts.

[0039] Specific preferred examples of the compound of the present invention include the compounds described in Examples 1 to 44 below, but the present invention is not construed as being limited thereto. The compounds described in Examples 1 to 44 are referred to as Compounds 1 to 44, respectively.

[0040] Among the above, Compound 3, Compound 8, Compound 10, Compound 11, Compound 13, Compound 14, Compound 15, Compound 16, Compound 17, Compound 19, Compound 20, Compound 21, Compound 22, Compound 23, Compound 24, Compound 27, Compound 28, Compound 29, Compound 32, Compound 33, Compound 34, Compound 37, Compound 38, Compound 39, Compound 42, Compound 43, and Compound 44 are preferred.

[0041] <Production Method> The production method of the compound of the present invention will be described. The compound of the present invention can be produced by combining known methods, for example, according to the production method shown below.

[0042] [Manufacturing method 1] "During the ceremony, R ａ and R ｂ represents a leaving group; R １ , R ２ , R ３ , R ４ , R ５ , R ６ , R ７ , and R ８ has the same meaning as above." Examples of leaving groups include a chloro group, a fluoro group, a bromo group, a trichloromethoxy group, a 4-nitro-phenoxy group, a 2,4-dinitrophenoxy group, a 2,4,6-trichlorophenoxy group, a pentafluorophenoxy group, a 2,3,5,6-tetrafluorophenoxy group, an imidazolyl group, a triazolyl group, a 3,5-dioxo-4-methyl-1,2,4-oxadiazolidyl group, and an N-hydroxysuccinimidyl group.

[0043] (1-1) Known examples of compounds of formula [3] include 1,1'-carbonyldiimidazole, 4-nitrophenyl chloroformate, triphosgene, and phosgene. Compounds of formula [4] can be produced by reacting compounds of formula [2] with compounds of formula [3] in the presence or absence of a base. The solvent used in this reaction is not particularly limited as long as it does not affect the reaction, and examples include halogenated hydrocarbons, ethers, esters, amides, nitriles, sulfoxides, and aromatic hydrocarbons. These solvents may be used in combination. Preferred solvents include ethers, with tetrahydrofuran being more preferred. The amount of solvent used is not particularly limited, and may be 1 to 500 times (v / w) the amount of the compound of formula [2]. Examples of bases used in this reaction include inorganic bases and organic bases. The base is preferably an organic base, and specific examples include triethylamine, N,N-diisopropylethylamine, 4-methylmorpholine, pyridine, 1,8-diazabicyclo[5.4.0]-7-undecene, and N,N-dimethylaminopyridine. The amount of the base used may be 1 to 50 times, preferably 1 to 10 times, the molar amount of the compound of formula [2]. The amount of the compound of formula [3] used is not particularly limited, but may be 0.3 to 10 times (v / w) the amount of the compound of formula [2]. This reaction may be carried out at -30 to 150°C, preferably 0 to 100°C, for 5 minutes to 48 hours.

[0044] (1-2) Known examples of compounds of formula [5] include 2,2'-((2-diethylamino)ethyl)azanediyl)bis(ethan-1-ol), 2,2'-((2-dimethylamino)ethyl)azanediyl)bis(ethan-1-ol), and 2,2'-((3-(diethylamino)propyl)azanediyl)bis(ethan-1-ol). The compound of formula [1] can be produced by reacting a compound of formula [4] with a compound of formula [5] in the presence of a base. The solvent used in this reaction is not particularly limited as long as it does not affect the reaction, and examples include halogenated hydrocarbons, ethers, esters, amides, nitriles, sulfoxides, and aromatic hydrocarbons. These solvents may be used in combination. Preferred solvents include nitriles, and acetonitrile is more preferred. The amount of solvent used is not particularly limited, and may be 1 to 500 times (v / w) the amount of the compound of formula [4]. The base used in this reaction may be an inorganic base or an organic base. Specific examples include potassium carbonate, sodium carbonate, lithium carbonate, potassium phosphate, sodium phosphate, lithium phosphate, triethylamine, N,N-diisopropylethylamine, 4-methylmorpholine, pyridine, 1,8-diazabicyclo[5.4.0]-7-undecene, or N,N-dimethylaminopyridine. The amount of the base used may be 1 to 50 times, preferably 1 to 10 times, the molar amount of the compound of formula [4]. The amount of the compound of formula [5] used is not particularly limited, but may be 0.1 to 10 times (v / w) the amount of the compound of formula [4]. This reaction may be carried out at −30 to 150°C, preferably 0 to 100°C, for 5 minutes to 48 hours.

[0045] (1-3) As compounds of formula [5], for example, 2-((2-(diethylamino)ethyl)(ethyl)amino)ethan-1-ol and 2-((2-(diethylamino)ethyl)(isopropyl)amino)ethan-1-ol are known. The compound of formula [1] can be produced by reacting a compound of formula [4] with a compound of formula [6] in the presence of a base. This reaction may be carried out in accordance with production method (1-2).

[0046] [Manufacturing method 2] "During the ceremony, R c and R e represents a leaving group; R d and R g represents a hydrocarbon group having 1 to 12 carbon atoms or hydrogen; R f represents a hydrocarbon group having 1 to 18 carbon atoms; R h represents a hydrocarbon group having 1 to 12 carbon atoms; M represents an alkali metal, an alkaline earth metal, or hydrogen; R i represents -MgCl, -MgBr, -MgI or -Li; R 21 , R 23 , and R 25 has the same meaning as above." Examples of leaving groups include a chloro group, a fluoro group, a bromo group, a trichloromethoxy group, a 4-nitro-phenoxy group, a 2,4-dinitrophenoxy group, a 2,4,6-trichlorophenoxy group, a pentafluorophenoxy group, a 2,3,5,6-tetrafluorophenoxy group, an imidazolyl group, a triazolyl group, a 3,5-dioxo-4-methyl-1,2,4-oxadiazolidyl group, and an N-hydroxysuccinimidyl group.

[0047] (2-1) As a compound of formula [8], for example, potassium ethyl malonate is known. The compound of formula [9] can be produced by reacting the compound of formula [7] with the compound of formula [8] in the presence or absence of a base and in the presence or absence of anhydrous magnesium chloride. The solvent used in this reaction is not particularly limited as long as it does not affect the reaction, and examples thereof include halogenated hydrocarbons, ethers, esters, amides, nitriles, sulfoxides, and aromatic hydrocarbons. These solvents may be used in mixtures. Preferred solvents include nitriles, and acetonitrile is more preferred. The amount of solvent used is not particularly limited, and may be 1 to 500 times (v / w) the amount of the compound of formula [7]. The base used in this reaction may be an inorganic base or an organic base. Organic bases are preferred, and specific examples include triethylamine, N,N-diisopropylethylamine, 4-methylmorpholine, pyridine, 1,8-diazabicyclo[5.4.0]-7-undecene, and N,N-dimethylaminopyridine. The amount of the base used may be 1 to 50 times, preferably 1 to 10 times, the molar amount of the compound of formula [7]. The amount of the compound of formula [8] used is not particularly limited, but may be 0.1 to 10 times (v / w) the amount of the compound of formula [7]. This reaction may be carried out at -30 to 150°C, preferably 0 to 100°C, for 5 minutes to 48 hours.

[0048] (2-2) Ethyl 8-bromooctanoate, for example, is known as a compound of formula

[10] . A compound of formula [11A] can be produced by reacting a compound of formula [9] with a compound of formula

[10] in the presence or absence of a base. The solvent used in this reaction is not particularly limited as long as it does not affect the reaction, and examples thereof include halogenated hydrocarbons, ethers, esters, amides, nitriles, sulfoxides, and aromatic hydrocarbons. These solvents may be used in combination. Preferred solvents include alcohols, with ethanol and methanol being more preferred. The amount of solvent used is not particularly limited, and may be 1 to 500 times (v / w) the amount of the compound of formula [9]. The base used in this reaction may be an inorganic base or an organic base. Specific examples of the base include potassium hydroxide, sodium hydroxide, lithium hydroxide, potassium carbonate, sodium carbonate, lithium carbonate, potassium phosphate, sodium phosphate, lithium phosphate, triethylamine, N,N-diisopropylethylamine, 4-methylmorpholine, pyridine, 1,8-diazabicyclo[5.4.0]-7-undecene, and N,N-dimethylaminopyridine. The amount of the base used may be 0.1 to 50 times, preferably 1 to 10 times, the molar ratio of the compound of formula [9]. The amount of the compound of formula [8] used is not particularly limited, but may be 0.1 to 10 times (v / w) the molar ratio of the compound of formula [9]. This reaction may be carried out at -30 to 150°C, preferably 0 to 100°C, for 5 minutes to 48 hours.

[0049] (2-3) The compound of formula [11B] can be produced by hydrolysis of the compound of formula [11A] in the presence of a base or an acid. The solvent used in this reaction is not particularly limited as long as it does not affect the reaction, and examples include halogenated hydrocarbons, ethers, esters, amides, nitriles, sulfoxides, aromatic hydrocarbons, and water. These solvents may be used in combination. The amount of solvent used is not particularly limited, and may be 1 to 500 times (v / w) the amount of the compound of formula [11A]. The base used in this reaction can be an inorganic or organic base. Inorganic bases are preferred, and specific examples include potassium hydroxide, sodium hydroxide, lithium hydroxide, potassium carbonate, sodium carbonate, lithium carbonate, potassium phosphate, sodium phosphate, and lithium phosphate. The acid used in this reaction can be an inorganic or organic acid. Inorganic acids are preferred, and specific examples include hydrochloric acid, hydrobromic acid, iodic acid, sulfuric acid, and phosphoric acid. The amount of the base used may be 0.1 to 50 times, preferably 1 to 10 times, the molar amount of the compound of formula [11A]. The amount of the acid used may be 0.01 to 50 times, preferably 1 to 10 times, the molar amount of the compound of formula [11A]. This reaction may be carried out at -30 to 150°C, preferably 0 to 100°C, for 5 minutes to 48 hours.

[0050] (2-4) For example, hexyl magnesium bromide is known as a compound of formula

[13] . The compound of formula [11B] can be produced by reacting a compound of formula

[12] with a compound of formula

[13] . The solvent used in this reaction is not particularly limited as long as it does not affect the reaction, and examples include halogenated hydrocarbons, ethers, amides, and aromatic hydrocarbons. These solvents may be used in combination. Preferred solvents include ethers, with tetrahydrofuran being more preferred. The amount of the solvent used is not particularly limited, but may be 1 to 500 times (v / w) the amount of the compound of formula

[12] . The amount of the compound of formula

[13] used is not particularly limited, but may be 0.8 to 10 times (v / w) the amount of the compound of formula

[12] . This reaction may be carried out at -30 to 150°C, preferably 0 to 100°C, for 5 minutes to 48 hours.

[0051] (2-5) Known examples of compounds of formula

[14] include 2-butyl-1-octanol and 2-pentyl-1-heptanol. Compounds of formula [11C] can be produced by reacting compounds of formula [11B] with compounds of formula

[14] in the presence or absence of an acid, in the presence or absence of a condensing agent or acid halide, and in the presence or absence of a base. The solvent used in this reaction is not particularly limited as long as it does not affect the reaction, and examples include halogenated hydrocarbons, ethers, esters, amides, nitriles, sulfoxides, and aromatic hydrocarbons. These solvents may be used in combination. Preferred solvents include aromatic hydrocarbons and ethers, with toluene and tetrahydrofuran being more preferred. The amount of solvent used is not particularly limited, but may be 1 to 500 times (v / w) the amount of the compound of formula [11B]. The acid used in this reaction can be an inorganic or organic acid. The acid is preferably a sulfonic acid, and specific examples include sulfuric acid, 4-toluenesulfonic acid, methanesulfonic acid, etc. Condensing agents used in this reaction include, for example, carbodiimides such as N,N'-dicyclohexylcarbodiimide and 1-ethyl-3-(3-dimethylaminopropyl)carbodiimide, carbonyls such as carbonyldiimidazole, acid azides such as diphenylphosphoryl azide, acid cyanides such as diethylphosphoryl cyanide, 2-ethoxy-1-ethoxycarbonyl-1,2-dihydroquinoline, and uroniums such as O-benzotriazol-1-yl-1,1,3,3-tetramethyluronium hexafluorophosphate and O-(7-azabenzotriazol-1-yl)-1,1,3,3-tetramethyluronium hexafluorophosphate. The acid halide used in this reaction includes, for example, carboxylic acid halides such as acetyl chloride and trifluoroacetyl chloride, sulfonic acid halides such as methanesulfonyl chloride and tosyl chloride, chloroformates such as ethyl chloroformate and isobutyl chloroformate, etc. The base used in this reaction includes an inorganic base or an organic base.The base is preferably an organic base, and specific examples include triethylamine, N,N-diisopropylethylamine, 4-methylmorpholine, pyridine, 1,8-diazabicyclo[5.4.0]-7-undecene, and N,N-dimethylaminopyridine. The amount of the base used may be 1 to 50 times, preferably 1 to 10 times, the molar amount of the compound of formula [11B]. The amount of the compound of formula

[14] used is not particularly limited, but may be 0.8 to 10 times (v / w) the amount of the compound of formula [11B]. This reaction may be carried out at -30 to 150°C, preferably 0 to 100°C, for 5 minutes to 48 hours.

[0052] (2-6) The compound of formula [2A] can be produced by a reduction reaction of the compound of formula [11C] in the presence of a reducing agent. The solvent used in this reaction is not particularly limited as long as it does not affect the reaction, and examples thereof include halogenated hydrocarbons, ethers, esters, amides, nitriles, alcohols, sulfoxides, aromatic hydrocarbons, and water. These solvents may be used in combination. The amount of the solvent used is not particularly limited, and may be 1 to 500 times (v / w) the amount of the compound of formula [11A]. Examples of reducing agents used in this reaction include boron compounds such as sodium borohydride. The amount of the reducing agent used may be 0.1 to 50 times, preferably 1 to 10 times, the molar amount of the compound of formula [11A]. This reaction may be carried out at −30 to 150°C, preferably 0 to 100°C, for 5 minutes to 48 hours.

[0053] [Manufacturing method 3] "During the ceremony, R 31 , R 32 , R 35 , and R 36 has the same meaning as above.

[0054] (3-1) Known examples of compounds of formula [2A] include 1-heptanol, 2-(hexylthio)ethan-1-ol, and 8-(methylthio)octan-1-ol. Compounds of formula

[17] can be produced by reacting compounds of formula

[15] with compounds of formula

[16] in the presence or absence of an acid, in the presence or absence of a condensing agent or an acid halide, and in the presence or absence of a base. This reaction can be carried out in accordance with Production Method (2-5).

[0055] (3-2) The compound of formula [2B] can be produced by reducing the compound of formula

[17] in the presence of a reducing agent. This reaction can be carried out in accordance with the production method (2-6).

[0056] [Manufacturing method 4] "During the ceremony, R j and R k represents a leaving group; R 1 , R 2 , R 3 , R 4 and R 8 has the same meaning as above." Examples of leaving groups include a chloro group, a fluoro group, a bromo group, a trichloromethoxy group, a 4-nitro-phenoxy group, a 2,4-dinitrophenoxy group, a 2,4,6-trichlorophenoxy group, a pentafluorophenoxy group, a 2,3,5,6-tetrafluorophenoxy group, an imidazolyl group, a triazolyl group, a 3,5-dioxo-4-methyl-1,2,4-oxadiazolidyl group, and an N-hydroxysuccinimidyl group.

[0057] (4-1) Known examples of compounds of formula

[19] include 2-chloro-N,N-diethylethan-1-amine, 3-chloro-N,N-diethylpropan-1-amine, and 2-bromo-N,N-diethylethan-1-amine. The compound of formula [5] can be produced by reacting a compound of formula

[18] with a compound of formula

[19] in the presence or absence of a base. The solvent used in this reaction is not particularly limited as long as it does not affect the reaction, and examples include alcohols, halogenated hydrocarbons, ethers, esters, amides, nitriles, sulfoxides, aromatic hydrocarbons, and water. These solvents may also be used in combination. The amount of solvent used is not particularly limited, and may be 1 to 500 times (v / w) the amount of the compound of formula

[18] . The base used in this reaction may be an inorganic base or an organic base. The amount of the base used may be 1 to 10,000 times, preferably 1 to 5,000 times, the molar amount of the compound of formula

[18] . The amount of the compound of formula

[19] used is not particularly limited, but may be 1 to 10 times (v / w) the amount of the compound of formula

[18] . This reaction may be carried out at -30 to 150°C, preferably 0 to 100°C, for 5 minutes to 48 hours.

[0058] (4-2) Known examples of compounds of formula

[20] include 2-bromo-1-ethanol and 3-bromo-1-propanol. The compound of formula [5] can be produced by reacting the compound of formula

[20] with the compound of formula

[21] in the presence or absence of a base. The solvent used in this reaction is not particularly limited as long as it does not affect the reaction, and examples include alcohols, halogenated hydrocarbons, ethers, esters, amides, nitriles, sulfoxides, aromatic hydrocarbons, and water. These solvents may be used in combination. The amount of the solvent used is not particularly limited, and may be 1 to 500 times (v / w) the amount of the compound of formula

[20] . The base used in this reaction may be an inorganic base or an organic base. The amount of the base used may be 1 to 10,000 times, preferably 1 to 5,000 times, the amount of the compound of formula

[20] by mole. The amount of the compound of formula

[21] used is not particularly limited, but may be 1 to 10 times (v / w) the amount of the compound of formula

[20] . This reaction may be carried out at −30 to 150° C., preferably 0 to 100° C., for 5 minutes to 48 hours.

[0059] In the compounds used in the above-mentioned production methods, when isomers (e.g., optical isomers, geometric isomers, tautomers, etc.) exist, these isomers can also be used. In addition, when solvates, hydrates, and various forms of crystals exist, these solvates, hydrates, and various forms of crystals can also be used.

[0060]

[0033] In the compounds used in the above-mentioned production methods, for example, compounds having an amino group, a hydroxyl group, or a carboxyl group can have these groups protected in advance with a conventional protecting group, and after the reaction, these protecting groups can be removed by a method known per se. The compounds obtained by the above-mentioned production methods can be derived into other compounds by subjecting them to a reaction known per se, such as condensation, addition, oxidation, reduction, rearrangement, substitution, halogenation, dehydration, or hydrolysis, or by an appropriate combination of these reactions.

[0061] <Lipid particles> In the present invention, lipid particles containing the compound of the present invention or a salt thereof can be prepared. When preparing lipid particles, in addition to the compound of the present invention, at least one lipid selected from the group consisting of sterols and lipids having nonionic hydrophilic polymer chains can be used. The lipid particles can further contain a neutral lipid. The lipid particles can further contain a nucleic acid.

[0062] In the lipid particles of the present invention, the amount of the compound of the present invention is preferably 20 mol % to 80 mol %, more preferably 35 mol % to 70 mol %, and even more preferably 40 mol % to 65 mol %, relative to the total lipid amount.

[0063] <Sterol> The lipid particles of the present invention preferably contain a sterol. By containing a sterol in the lipid particles of the present invention, membrane fluidity can be reduced and a stabilizing effect of the lipid particles can be obtained. The sterol is not particularly limited, but examples thereof include cholesterol, phytosterol (sitosterol), stigmasterol, fucosterol, spinasterol, brassicasterol, etc.), ergosterol, cholestanone, cholestenone, coprostanol, cholesteryl-2'-hydroxyethyl ether, cholesteryl-4'-hydroxybutyl ether, etc. Among these, cholesterol is preferred. In the lipid particles of the present invention, the amount of sterol blended is preferably 10 mol% to 60 mol%, more preferably 20 mol% to 55 mol%, and even more preferably 25 mol% to 50 mol%, relative to the total lipid amount.

[0064] <Neutral lipid> The lipid particle of the present invention may contain a neutral lipid. The neutral lipid is not particularly limited, but may include phosphatidylcholine, phosphatidylethanolamine, sphingomyelin, ceramide, etc., and phosphatidylcholine is preferred. In addition, the neutral lipid may be used alone or in combination with a plurality of different neutral lipids.

[0065] Examples of phosphatidylcholines include, but are not limited to, soybean lecithin (SPC), hydrogenated soybean lecithin (HSPC), egg yolk lecithin (EPC), hydrogenated egg yolk lecithin (HEPC), dimyristoylphosphatidylcholine (DMPC), dipalmitoylphosphatidylcholine (DPPC), distearoylphosphatidylcholine (DSPC), 1-palmitoyl-2-oleoylphosphatidylcholine (POPC), and dioleoylphosphatidylcholine (DOPC).

[0066] The phosphatidylethanolamine is not particularly limited, and examples thereof include dimyristoylphosphatidylethanolamine (DMPE), dipalmitoylphosphatidylethanolamine (DPPE), distearoylphosphatidylethanolamine (DSPE), dioleoylphosphatidylethanolamine (DOPE), dilinoleoylphosphatidylethanolamine (DLoPE), diphytanoylphosphatidylethanolamine (D(Phy)PE), 1-palmitoyl-2-oleoylphosphatidylethanolamine (POPE), ditetradecylphosphatidylethanolamine, dihexadecylphosphatidylethanolamine, dioctadecylphosphatidylethanolamine, and diphytanylphosphatidylethanolamine.

[0067] Examples of sphingomyelin include, but are not limited to, egg yolk-derived sphingomyelin, milk-derived sphingomyelin, etc. Examples of ceramide include, but are not limited to, egg yolk-derived ceramide, milk-derived ceramide, etc.

[0068] In the lipid particles of the present invention, the blending amount of the neutral lipid is preferably 0 mol % or more and 55 mol % or less based on the total amount of the constituent lipid components.

[0069] <Lipids with nonionic hydrophilic polymer chains> The lipid particles of the present invention may contain lipids with nonionic hydrophilic polymer chains in the oil phase. In the present invention, by containing lipids with nonionic hydrophilic polymer chains in the oil phase, the dispersion stabilization effect of the lipid particles can be obtained. Examples of nonionic hydrophilic polymers include, but are not limited to, nonionic vinyl polymers, nonionic polyamino acids, nonionic polyesters, nonionic polyethers, nonionic natural polymers, nonionic modified natural polymers, and block polymers or graft copolymers containing two or more of these polymers as structural units. Among these nonionic hydrophilic polymers, preferred are nonionic polyethers, nonionic polyesters, nonionic polyamino acids, or nonionic synthetic polypeptides, more preferred are nonionic polyethers or nonionic polyesters, even more preferred are nonionic polyethers or nonionic monoalkoxy polyethers, and particularly preferred is polyethylene glycol (polyethylene glycol will also be referred to as PEG hereinafter).

[0070] Lipids having a nonionic hydrophilic polymer chain include, but are not limited to, PEG-modified phosphoethanolamine, diacylglycerol PEG derivatives, monoacylglycerol PEG derivatives, dialkylglycerol PEG derivatives, cholesterol PEG derivatives, and ceramide PEG derivatives. Among these, monoacylglycerol PEG or diacylglycerol PEG is preferred. The weight-average molecular weight of the PEG chain of the nonionic hydrophilic polymer derivative is preferably 500 to 5,000, more preferably 750 to 3,000. The nonionic hydrophilic polymer chain may be branched or may have a substituent such as a hydroxymethyl group.

[0071] In the lipid particles of the present invention, the blending amount of the lipid having a nonionic hydrophilic polymer chain is preferably 0.25 mol % to 12 mol %, more preferably 0.5 mol % to 6 mol %, and even more preferably 1 mol % to 3 mol %, relative to the total lipid amount.

[0072] <Nucleic Acid> The lipid particles of the present invention may contain nucleic acid. Examples of nucleic acids include plasmids, single-stranded DNA, double-stranded DNA, siRNA (small interfering RNA), miRNA (micro RNA), mRNA, antisense oligonucleotides (also known as ASO), ribozymes, aptamers, saRNA, sgRNA, etc., and any of these may be contained. Modified nucleic acids may also be contained. RNA is particularly preferred as the nucleic acid, and RNA having 5 to 20,000 bases is preferred. In the lipid particles of the present invention, the mass ratio of lipid to nucleic acid is preferably 2 to 1,000, more preferably 3 to 500, even more preferably 5 to 200, and particularly preferably 5 to 100.

[0073] <Method of Manufacturing Lipid Particles> A method of manufacturing lipid particles of the present invention will now be described. The method of manufacturing lipid particles is not limited, but lipid particles can be manufactured by dissolving all or some of the oil-soluble components constituting the lipid particles in an organic solvent or the like to form an oil phase, dissolving the water-soluble components in water to form an aqueous phase, and mixing the oil phase and aqueous phase. A micromixer may be used for mixing, or emulsification may be performed using an emulsifier such as a homogenizer, an ultrasonic emulsifier, a high-pressure injection emulsifier, or the like. Alternatively, lipid-containing solutions may be dried under reduced pressure using an evaporator or the like, or spray-dried using a spray dryer or the like to prepare a dried mixture containing lipids, and this mixture is added to an aqueous solvent and further emulsified using the aforementioned emulsifier or the like.

[0074] An example of a method for producing lipid particles containing nucleic acid includes the following steps: (a) dissolving the components of lipid particles containing the compound of the present invention in an organic solvent to obtain an oil phase; (b) mixing the oil phase obtained in step (a) with an aqueous phase containing nucleic acid; (c) diluting the mixture containing the oil phase and aqueous phase obtained in step (b) to obtain a dispersion of nucleic acid-lipid particles; and (d) removing the organic solvent from the dispersion of nucleic acid-lipid particles obtained in step (c).

[0075] In step (a), the components of lipid particles containing the compound of the present invention are dissolved in an organic solvent (an alcohol such as ethanol, an ester, etc.). The total lipid concentration is not particularly limited, but is generally 1 mmol / L to 100 mmol / L, preferably 5 mmol / L to 50 mmol / L, and more preferably 10 mmol / L to 30 mmol / L.

[0076] In step (b), the aqueous phase can be obtained by dissolving nucleic acids (e.g., siRNA, mRNA, antisense nucleic acids, etc.) in water or a buffer solution. Components such as antioxidants can be added as needed. The mixing ratio (volume ratio) of the aqueous phase to the oil phase is preferably 5:1 to 1:1, more preferably 4:1 to 2:1.

[0077] In step (b), the mixture can be diluted with water or a buffer solution (such as phosphate buffered saline (PBS)).

[0078] In step (d), the method for removing the organic solvent from the dispersion of nucleic acid-lipid particles is not particularly limited, and any common method can be used. For example, the organic solvent can be removed by dialysis using a solution such as phosphate-buffered saline or sucrose-Tris buffer.

[0079] The lipid particles can be sized as needed. The sizing method is not particularly limited, but the particle size can be reduced using an extruder or the like. In addition, the dispersion containing the lipid particles of the present invention can be frozen or freeze-dried by a general method.

[0080] <About Lipid Particles> In the present invention, lipid particles refer to particles composed of lipids, and include compositions having any structure selected from lipid aggregates in which lipids are aggregated, micelles, and liposomes, but the structure of the lipid particles is not limited to these as long as the composition contains lipids. Liposomes include liposomes with a lipid bilayer structure and an aqueous phase inside, with a single-layer bilayer membrane, and multilayer liposomes with multiple layers stacked on top of each other. Either type of liposome may be included in the present invention.

[0081] The morphology of lipid particles can be confirmed by electron microscope observation or X-ray structural analysis.For example, by using a cryo-transmission electron microscope (cryo-TEM) method, it can be confirmed whether the lipid particles have a lipid bilayer structure (lamellar structure) and an inner water layer, like liposomes, or whether the particles have a core with high electron density inside and a structure packed with lipids and other components.Small-angle X-ray scattering (SAXS) measurement can also be used to confirm whether the lipid particles have a lipid bilayer structure (lamellar structure).

[0082] The particle size of the lipid particles of the present invention is not particularly limited, but is preferably 10 to 1,000 nm, more preferably 30 to 500 nm, and even more preferably 50 to 250 nm. The particle size of the lipid particles can be measured by a general method (e.g., dynamic light scattering method, laser diffraction method, etc.).

[0083] <Use of Lipid Particles> As an example of the use of lipid particles in the present invention, nucleic acid (e.g., RNA) can be introduced into cells by introducing lipid particles containing nucleic acid into cells. Furthermore, when the lipid particles of the present invention contain nucleic acid having pharmaceutical uses, the lipid particles can be administered to a living body as a nucleic acid drug.

[0084] When the lipid particles of the present invention are used as nucleic acid medicines, the lipid particles of the present invention can be administered to living bodies alone or mixed with pharmaceutically acceptable administration vehicles (for example, physiological saline or phosphate buffer).The concentration of lipid particles in the mixture with pharmaceutically acceptable carriers is not particularly limited, and can generally be 0.05% by mass to 90% by mass.In addition, the nucleic acid medicines containing the lipid particles of the present invention may also contain other pharmaceutically acceptable additives, such as pH adjusting buffers, osmotic pressure adjusting agents, etc.

[0085] The administration route of the nucleic acid drug containing the lipid particles of the present invention is not particularly limited, and can be administered by any method.Administration methods include oral administration and parenteral administration (intra-articular administration, intravenous administration, intra-arterial administration, subcutaneous administration, intradermal administration, intravitreal administration, intraperitoneal administration, intramuscular administration, intravaginal administration, intravesical administration, intrathecal administration, pulmonary administration, rectal administration, colonic administration, buccal administration, nasal administration, intracisternal administration, inhalation, etc.).Parenteral administration is preferred, and the preferred administration methods are intravenous injection, subcutaneous injection, intradermal injection, or intramuscular injection.The nucleic acid drug containing the lipid particles of the present invention can also be administered by direct injection into the diseased site.

[0086] The dosage form of the lipid particles of the present invention is not particularly limited, but when administered orally, the lipid particles of the present invention can be combined with an appropriate excipient and used in the form of tablets, troches, capsules, pills, suspensions, syrups, etc. Furthermore, preparations suitable for parenteral administration can contain additives such as antioxidants, buffers, bacteriostatic agents, and isotonic sterile injections, suspending agents, solubilizing agents, thickening agents, stabilizers, or preservatives, as appropriate.

[0087] <Nucleic Acid Delivery Carrier> The lipid particles of the present invention are capable of retaining nucleic acids at a high encapsulation rate, and are therefore very useful as nucleic acid delivery carriers. According to the nucleic acid delivery carrier utilizing the present invention, for example, the obtained lipid particles can be mixed with nucleic acids or the like, and transfected in vitro or in vivo, thereby introducing nucleic acids or the like into cells. Furthermore, the nucleic acid delivery carrier utilizing the present invention is also useful as a nucleic acid delivery carrier for nucleic acid medicines. That is, the lipid particles of the present invention are useful as a composition for nucleic acid delivery in vitro or in vivo (preferably in vivo).

[0088] The present invention will now be described with reference to examples, but the present invention is not limited to these examples.

[0089] Unless otherwise specified, purification by column chromatography was performed using an ISOLERA automatic purification system (Biotage), a Purif-espoir-2 medium-pressure preparative purification system (Shoko Science Co., Ltd.), or a YFLC W-prep 2XY medium-pressure liquid chromatograph (Yamazen Corporation). Unless otherwise specified, the carrier used in silica gel column chromatography was Chromatorex Q-Pack SI 50 (Fuji Silysia Chemical Ltd.), or Hi-Flash Column W001, W002, W003, W004, or W005 (Yamazen Corporation). NH silica gel was Chromatorex Q-Pack NH 60 (Fuji Silysia Chemical Ltd.). NMR spectra were measured using tetramethylsilane as an internal standard on a Bruker AV300 (Bruker), Bruker AV400 (Bruker), or AVNEO400 (Bruker). All δ values ​​are expressed in ppm. MS spectra were measured using an ACQUITY SQD LC / MS System (Waters). clogP was calculated using ChemDraw Professional Version: 19.1.0.8 (PerkinElmer).

[0090] [Example 1] (1) To a mixture of potassium monoethyl malonate (60.1 g) and acetonitrile (400 mL), triethylamine (75.0 mL) and anhydrous magnesium chloride (40.0 g) were added under ice-cooling, and the mixture was stirred at room temperature for 2 hours. To the reaction mixture, heptanoyl chloride (25.0 g) was added dropwise under ice-cooling, and the mixture was stirred at room temperature for 2 hours. The solvent from the reaction mixture was evaporated under reduced pressure, and toluene (200 mL) was added. After evaporation under reduced pressure, toluene (100 mL) was added. 15% aqueous hydrochloric acid (250 mL) was added dropwise to the resulting residue under ice-cooling, and the organic layer was separated and washed with 15% aqueous hydrochloric acid (76 mL), followed by water (75 mL). Toluene (100 mL) was added, and the mixture was evaporated under reduced pressure to give ethyl 3-oxononanoate (35.2 g). 1H-NMR(CDCl3)δ: 4.20 (2H, q, J=8.0Hz), 3.43 (2H, s), 2.53 (2H, t, J=8.0Hz), 1.65-1.52 (2H, m), 1.26-1.21 (9H, m), 0.88 (3H, t, 8.0Hz).

[0091] (2) A solution of ethyl 3-oxononanoate (15.0 g) in ethanol (10 mL) was added to a 20% sodium ethoxide-ethanol solution (32 mL), followed by dropwise addition of ethyl 8-bromooctanoate (18.9 g) and stirring at 90°C for 4 hours. After the reaction mixture was cooled to room temperature, 33% aqueous sodium hydroxide (22.5 mL) was added and the mixture was stirred at room temperature for 1 hour. 15% aqueous hydrochloric acid (48 mL) was added to the reaction mixture and stirred at 60°C for 30 minutes. After the reaction mixture was cooled to 40°C, the organic layer was separated and the solvent was evaporated under reduced pressure. Ethyl acetate and water were added to the resulting residue, the organic layer was separated and the solvent was evaporated under reduced pressure. Hexane was added to the resulting residue, and the solid was collected by filtration, washed with hexane, and dried under reduced pressure to give 10-oxohexadecanoic acid (9.1 g). 1 H-NMR(CDCl3)δ: 2.45-2.28 (6H, m), 1.71-1.47 (6H, m), 1.40-1.20 (14H, m), 0.88 (3H, t, J=8.0Hz).

[0092] (3) To a mixture of 10-oxohexadecanoic acid (2.0 g), 2-hexyl-1-decanol (1.8 g), and toluene (20 mL), p-toluenesulfonic acid (0.07 g) was added and stirred at 130° C. for 3 hours. After the reaction mixture was cooled to room temperature, a 5% aqueous solution of sodium hydrogen carbonate was added, and the organic layer was separated, washed with water, dried over anhydrous magnesium sulfate, and the solvent was distilled off under reduced pressure to obtain 2-hexyldecyl 10-oxohexadecanoate (3.7 g). 1H-NMR(CDCl3)δ:3.97 (2H, d, J=5.6Hz), 2.38 (4H, t, J=7.6Hz), 2.29 (2H, t, J=7.6Hz), 1.65-1.50 (7H, m), 1.35-1.20 (38H, m), 0.92-0.83 (9H, m).

[0093] (4) To a mixture of 2-hexyldecyl 10-oxohexadecanoate (3.7 g) and toluene (18 mL), sodium borohydride (0.42 g) was added, and then methanol (3.7 mL) was added dropwise under ice-cooling, followed by stirring at the same temperature for 3 hours. 30% aqueous hydrochloric acid solution (18 mL) was added dropwise to the reaction mixture under ice-cooling, and the organic layer was separated, washed with water, dried over anhydrous magnesium sulfate, and evaporated under reduced pressure to obtain 2-hexyldecyl 10-hydroxyhexadecanoate (3.5 g). 1 H-NMR(CDCl3)δ:3.97 (2H, d, J=6.0Hz), 3.61-3.54 (1H, m), 2.30 (2H, t, J=7.6Hz), 1.65-1.56 (3H, m), 1.48-1.22 (46H, m), 0.92-0.83 (9H, m).

[0094] (5) To a mixture of 2-hexyldecyl 10-hydroxyhexadecanoate (3.6 g), triethylamine (3.1 mL), and tetrahydrofuran (36 mL), 4-nitrophenyl chloroformate (2.21 g) was added and stirred at room temperature for 4 hours. Water and ethyl acetate were added to the reaction mixture, and the organic layer was separated and washed with water, then dried over anhydrous sodium sulfate, and the solvent was distilled off under reduced pressure. The resulting residue was purified by silica gel column chromatography (ethyl acetate-hexane) to give 2-hexyldecyl 10-(((4-nitrophenoxy)carbonyl)oxy)hexadecanoate (4.5 g) as a colorless oil. 1H-NMR(CDCl3)δ:8.28 (2H, dd, J=7.2Hz, 2.1Hz), 7.39 (2H, dd, J=7.2Hz, 2.1Hz), 4.86-4.76 (1H, m), 3.97 (2H, d, J=5.7Hz), 2.30 (2H, t, J=7.2Hz), 1.74-1.20 (49H, m), 0.92-0.85 (9H, m).

[0095] (6) To a mixture of 2,2'-azanediylbis(ethan-1-ol) (2.0 g), 2-bromo-N,N-diethylethan-1-amine hydrobromide (7.4 g), and ethanol (40 mL), potassium carbonate (7.9 g) was added and the mixture was stirred under reflux for 8 hours. The reaction mixture was cooled to room temperature, and the insoluble materials were removed by filtration, followed by evaporation of the solvent under reduced pressure. The resulting residue was purified by silica gel column chromatography (ethyl acetate-hexane, NH silica gel) to afford 2,2'-((2-(diethylamino)ethyl)azanediyl)bis(ethan-1-ol) (2.3 g) as a pale yellow oil. 1 H-NMR(CDCl3)δ:3.58 (4H, t, J=5.4Hz), 2.70 (4H, t, J=5.4Hz), 2.67-2.48 (8H, m), 1.04 (6H, t, J=7.5Hz). MS m / z(M+H):205.

[0096] (7) To a mixture of 2-hexyldecyl 10-(((4-nitrophenoxy)carbonyl)oxy)hexadecanoate (2.00 g), 2,2'-((2-(diethylamino)ethyl)azanediyl)bis(ethan-1-ol) (1.85 g), triethylamine (1.27 mL), and tetrahydrofuran (20 mL), 4-dimethylaminopyridine (1.11 mg) was added and stirred at 80°C for 4 hours. The reaction mixture was cooled to room temperature, and water and ethyl acetate were added. The organic layer was separated, washed with a saturated aqueous sodium chloride solution, dried over anhydrous sodium sulfate, and the solvent was evaporated under reduced pressure. The obtained residue was purified by silica gel column chromatography (methanol-ethyl acetate) and silica gel column chromatography (ethyl acetate-hexane, NH silica gel) to obtain a colorless oily substance, bis(2-hexyldecyl) 16-(2-(diethylamino)ethyl)-10,22-dihexyl-12,20-dioxo-11,13,19,21-tetraoxa-16-azahentriacontanedioate (197 mg). 1 H-NMR(CDCl3)δ: 4.71-4.59 (2H, m), 4.21-4.08 (4H, m), 3.97 (4H, d, J=6.0Hz), 2.84 (4H, t, J=6.6Hz), 2.71-2.44 (8H, m), 2.29 (4H, t, MS m / z(M+H):1251. clogP:30.3764

[0097] [Example 2] A colorless oily substance, bis(2-butyloctyl) 16-(2-(diethylamino)ethyl)-10,22-dihexyl-12,20-dioxo-11,13,19,21-tetraoxa-16-azahentriacontanedioate, was obtained in the same manner as in Example 1, except that 2-butyl-1-octanol was used instead of 2-hexyl-1-decanol. 1H-NMR(CDCl3)δ: 4.72-4.58 (2H, m), 4.21-4.08 (4H, m), 3.97 (4H, d, J=6.0Hz), 2.84 (4H, t, J=6.6Hz), 2.73-2.43 (8H, m), 2.29 (4H, t, MS m / z(M+H):1139. clogP:26.1444

[0098] [Example 3] (1) To a mixture of 2,2'-azanediylbis(ethan-1-ol) (2.5 g), 3-chloro-N,N-diethylpropan-1-amine (4.6 g), and ethanol (25 mL), potassium carbonate (4.3 g) was added and the mixture was stirred under reflux for 6 hours. After the reaction mixture was cooled to room temperature, insoluble materials were removed by filtration, and the solvent was evaporated under reduced pressure. The resulting residue was purified by silica gel column chromatography (ethyl acetate-hexane, NH silica gel) to give 2,2'-((3-(diethylamino)propyl)azanediyl)bis(ethan-1-ol) (2.9 g) as a pale yellow oil. 1 H-NMR(CDCl3)δ: 3.62 (4H, t, J=5.2Hz), 2.26 (2H, t, J=6.0Hz), 2.61-2.49 (10H, m), 1.68-1.60 (2H, m), 1.04 (6H, t, J=7.2Hz). MS m / z(M+H):219.

[0099] (2) A colorless oily substance, bis(2-butyloctyl)16-(3-(diethylamino)propyl)-10,22-dihexyl-12,20-dioxo-11,13,19,21-tetraoxa-16-azahentriacontanedioate, was obtained in the same manner as in Example 1, except that 2-butyl-1-octanol was used instead of 2-hexyl-1-decanol, and 2,2'-((3-(diethylamino)propyl)azanediyl)bis(ethan-1-ol) was used instead of 2,2'-((2-(diethylamino)ethyl)azanediyl)bis(ethan-1-ol). 1 H-NMR(CDCl3)δ: 4.72-4.60 (2H, m), 4.23-4.07 (4H, m), 3.97 (4H, d, J=6.0Hz), 2.81 (4H, t, J=6.6Hz), 2.72-2.37 (8H, m), 2.29 (4H, t, MS m / z(M+H): 1153. clogP: 26.389

[0100] [Example 4] A colorless oily substance, bis(3-pentyloctyl)16-(2-(diethylamino)ethyl)-10,22-dihexyl-12,20-dioxo-11,13,19,21-tetraoxa-16-azahentriacontanedioate, was obtained in the same manner as in Example 1, except that 3-pentyloctan-1-ol was used instead of 2-hexyl-1-decanol. 1H-NMR(CDCl3)δ: 4.72-4.58 (2H, m), 4.21-4.11 (4H, m), 3.97 (4H, t, J=7.2Hz), 2.84 (4H, t, J=6.6Hz), 2.71-2.64 (2H, m), 2.56-2.46 (6H, MS m / z(M+H):1167.

[0101] [Example 5] A colorless oily substance, bis(2-pentylheptyl)16-(2-(diethylamino)ethyl)-10,22-dihexyl-12,20-dioxo-11,13,19,21-tetraoxa-16-azahentriacontanedioate, was obtained in the same manner as in Example 1, except that 2-pentylheptan-1-ol was used instead of 2-hexyl-1-decanol. 1 H-NMR(CDCl3)δ: 4.72-4.58 (2H, m), 4.21-4.08 (4H, m), 3.97 (4H, d, J=6.0Hz), 2.84 (4H, t, J=6.6Hz), 2.71-2.64 (2H, m), 2.56-2.46 (6H, MS m / z(M+H):1139. clogP:26.1444

[0102] [Example 6] A colorless oily substance, bis(2-hexyloctyl)16-(2-(diethylamino)ethyl)-10,22-dihexyl-12,20-dioxo-11,13,19,21-tetraoxa-16-azahentriacontanedioate, was obtained in the same manner as in Example 1, except that 2-hexyloctan-1-ol was used instead of 2-hexyl-1-decanol. 1H-NMR(CDCl3)δ: 4.72-4.58 (2H, m), 4.21-4.08 (4H, m), 3.97 (4H, d, J=6.0Hz), 2.84 (4H, t, J=6.6Hz), 2.71-2.64 (2H, m), 2.56-2.46 (6H, MS m / z(M+H):1195. clogP: 28.2604

[0103] [Example 7] (1) A mixture of 10-methoxy-10-oxodecanoic acid (47.6 g), thionyl chloride (47.6 mL), and N,N-dimethylformamide (0.1 mL) was stirred under reflux for 1 hour. The solvent was evaporated under reduced pressure to give methyl 10-chloro-10-oxodecanoate (59.7 g) as a brown oil. 1 H-NMR(CDCl3)δ:3.67 (3H, s), 2.88 (2H, t, J=7.2Hz), 2.30 (2H, t, J=7.2Hz), 1.75-1.57 (4H, m), 1.38-1.25 (8H, m).

[0104] (2) To a suspension of zinc(II) chloride (13.6 g) in tetrahydrofuran (300 mL), a 1.0 mol / L pentylmagnesium bromide-tetrahydrofuran solution (200 mL) was added dropwise at -78°C, and the mixture was warmed to 0°C and stirred at the same temperature for 30 minutes. Tetrakis(triphenylphosphine)palladium(0) (2.9 g) was added to the reaction mixture under ice cooling, and then methyl 10-chloro-10-oxodecanoate (24.9 g) was added dropwise at the same temperature and stirred at the same temperature for 1 hour. To the reaction mixture, 1.0 mol / L aqueous hydrochloric acid solution (100 mL), hexane (100 mL), and ethyl acetate (100 mL) were added, and the organic layer was separated, washed with saturated aqueous sodium chloride solution (100 mL), dried over anhydrous sodium sulfate, and the solvent was evaporated under reduced pressure. The resulting residue was purified by silica gel column chromatography (ethyl acetate-hexane) to obtain methyl 10-oxopentadecanoate (19.8 g) as a white solid. 1 H-NMR(CDCl3)δ:3.67 (3H, s), 2.38 (4H, t, J=7.2Hz), 2.30 (2H, t, 7.2Hz), 1.65-1.49 (6H, m), 1.35-1.20 (12H, m), 0.88 (3H, t, J=7.2Hz).

[0105] (3) To a mixture of methyl 10-oxopentadecanoate (5.3 g) and 2-hexyldecan-1-ol (7.1 g), tetraisopropyl orthotitanate (0.55 g) was added and stirred for 2 hours at 110° C. After the reaction mixture was cooled to room temperature, water (0.5 mL) was added, followed by stirring at room temperature for 15 minutes. The mixture was purified by silica gel column chromatography (ethyl acetate-hexane) to obtain 2-hexyldecyl 10-oxopentadecanoate (9.2 g) as a colorless oil. 1 H-NMR(CDCl3)δ:3.97 (2H, d, J=5.6Hz), 2.38 (4H, t, J=7.6Hz), 2.29 (2H, t, J=7.6Hz), 1.65-1.50 (7H, m), 1.35-1.20 (36H, m), 0.92-0.83 (9H, m).

[0106] Sodium borohydride (1.0 g) was added to a mixture of 2-hexyldecyl 10-oxopentadecanoate (9.2 g), methanol (36 mL), and tetrahydrofuran (36 mL) under ice-cooling, and the mixture was stirred at the same temperature for 1 hour. Water (80 mL), 1.0 mol / L aqueous hydrochloric acid (35 mL), and hexane (40 mL) were added to the reaction mixture, and the organic layer was separated. The mixture was washed with saturated aqueous sodium chloride (20 mL) and dried over anhydrous magnesium sulfate, and the solvent was removed under reduced pressure. The resulting residue was purified by silica gel column chromatography (ethyl acetate-hexane) to yield 2-hexyldecyl 10-hydroxypentadecanoate (7.2 g) as a colorless oil. 1 H-NMR(CDCl3)δ:3.97 (2H, d, J=6.0Hz), 3.61-3.54 (1H, m), 2.30 (2H, t, J=7.6Hz), 1.65-1.56 (3H, m), 1.48-1.22 (44H, m), 0.92-0.83 (9H, m).

[0107] To a mixture of 2-hexyldecyl 10-hydroxypentadecanoate (2.1 g), triethylamine (2.4 mL), and tetrahydrofuran (21 mL), 4-nitrophenyl chloroformate (1.8 g) was added and stirred at room temperature for 4 hours. Water (60 mL) and hexane (60 mL) were added to the reaction mixture, and the organic layer was separated and washed with water, dried over anhydrous sodium sulfate, and the solvent was distilled off under reduced pressure. The resulting residue was purified by silica gel column chromatography (ethyl acetate-hexane) to give 2-hexyldecyl 10-(((4-nitrophenoxy)carbonyl)oxy)pentadecanoate (2.7 g) as a colorless oil. 1 H-NMR(CDCl3)δ:8.28 (2H, dd, J=7.2Hz, 2.1Hz), 7.39 (2H, dd, J=7.2Hz, 2.1Hz), 4.86-4.76 (1H, m), 3.97 (2H, d, J=5.7Hz), 2.30 (2H, t, J=7.2Hz), 1.74-1.20 (47H, m), 0.92-0.85 (9H, m).

[0108] To a mixture of 2-hexyldecyl 10-(((4-nitrophenoxy)carbonyl)oxy)pentadecanoate (2.00 g), 2,2'-((2-(diethylamino)ethyl)azanediyl)bis(ethan-1-ol) (1.89 g), triethylamine (1.29 mL), and tetrahydrofuran (10 mL), 4-dimethylaminopyridine (1.13 g) was added and stirred at 80°C for 6 hours. The reaction mixture was cooled to room temperature, and water and ethyl acetate were added. The organic layer was separated, washed with a saturated aqueous sodium chloride solution, dried over anhydrous sodium sulfate, and the solvent was evaporated under reduced pressure. The obtained residue was purified by silica gel column chromatography (methanol-ethyl acetate) and silica gel column chromatography (ethyl acetate-hexane, NH silica gel) to obtain a colorless oily substance, bis(2-hexyldecyl) 16-(2-(diethylamino)ethyl)-12,20-dioxo-10,22-dipentyl-11,13,19,21-tetraoxa-16-azahentriacontanedioate (0.12 g). 1 H-NMR(CDCl3)δ: 4.72-4.58 (2H, m), 4.21-4.08 (4H, m), 3.97 (4H, d, J=6.0Hz), 2.84 (4H, t, J=6.6Hz), 2.71-2.64 (2H, m), 2.56-2.46 (6H, MS m / z(M+H):1223. clogP: 29.3184

[0109] [Example 8] (1) To a solution of glutaric anhydride (27.3 g) in tetrahydrofuran (273 mL), a 1.0 mol / L hexylmagnesium bromide-tetrahydrofuran solution (200 mL) was added dropwise under ice-cooling, and the mixture was stirred at the same temperature for 1 hour. To the reaction mixture, a 2 mol / L aqueous hydrochloric acid solution (240 mL) was added under ice-cooling, followed by ethyl acetate (270 mL). The organic layer was separated, washed with water and saturated aqueous sodium chloride, dried over anhydrous magnesium sulfate, and the solvent was evaporated under reduced pressure. The resulting residue was purified by silica gel column chromatography (ethyl acetate-hexane), and then hexane (10 mL) was added. The solid was collected by filtration, washed with hexane, and dried under reduced pressure to yield 5-oxoundecanoic acid (16.0 g) as a white solid. 1 H-NMR(CDCl3)δ:2.50 (2H, t, J=7.2Hz), 2.40 (4H, t, J=7.2Hz), 2.02-1.80 (2H, m), 1.63-1.48 (2H, m), 1.37-1.20 (6H, m), 0.88 (3H, t, J=6.6Hz).

[0110] (2) To a mixture of 5-oxoundecanoic acid (3.0 g), 2-butyloctan-1-ol (2.5 g), and toluene (6.0 mL), p-toluenesulfonic acid (0.14 g) was added and stirred for 2 hours at 100° C. The reaction mixture was cooled to room temperature and then purified by silica gel column chromatography (ethyl acetate-hexane) to obtain 2-butyloctyl 5-oxoundecanoate (5.0 g) as a colorless oil. 1 H-NMR(CDCl3)δ:3.97 (2H, d, J=5.1Hz), 2.47 (2H, t, J=7.2Hz), 2.39 (2H, t, J=7.2Hz), 2.33 (2H, t, J=7.2Hz), 1.95-1.83 (2H, m), 1.66-1.49 (3H, m), 1.36-1.20 (22H, m), 0.92-0.82 (9H, m).

[0111] To a mixture of 2-butyloctyl 5-oxoundecanoate (7.3 g), toluene (30 mL), and methanol (30 mL), sodium borohydride (0.90 g) was added under ice-cooling, and the mixture was stirred at the same temperature for 1 hour. Water (30 mL), 2.0 mol / L aqueous hydrochloric acid (30 mL), and ethyl acetate (30 mL) were added to the reaction mixture under ice-cooling, and the organic layer was separated. After washing with saturated aqueous sodium chloride and drying over anhydrous sodium sulfate, the solvent was evaporated under reduced pressure. The resulting residue was purified by silica gel column chromatography (ethyl acetate-hexane) to yield 2-butyloctyl 5-hydroxyundecanoate (6.9 g) as a colorless oil. 1 H-NMR(CDCl3)δ:3.97 (2H, d, J=5.7Hz), 3.65-3.53 (1H, m), 2.35 (2H, t, J=7.2Hz), 1.87-1.20 (32H, m), 0.92-0.84 (9H, m).

[0112] (3) To a mixture of 2-butyloctyl 5-hydroxyundecanoate (1.62 g), triethylamine (2.38 mL), and tetrahydrofuran (16 mL), 4-nitrophenyl chloroformate (1.71 g) was added and stirred at room temperature for 4 hours. Water and ethyl acetate were added to the reaction mixture, and the organic layer was separated and washed with water, then dried over anhydrous sodium sulfate, and the solvent was distilled off under reduced pressure. The resulting residue was purified by silica gel column chromatography (ethyl acetate-hexane) to give 2-butyloctyl 5-(((4-nitrophenoxy)carbonyl)oxy)undecanoate (1.99 g) as a colorless oil. 1 H-NMR(CDCl3)δ:8.28 (2H, d, J=9.3Hz), 7.39 (2H, d, J=9.3Hz), 4.88-4.77 (1H, m), 3.99 (2H, d, J=6.0Hz), 2.41-2.31 (2H, m), 1.80-1.48 (7H, m), 1.44-1.20 (24H, m), 0.92-0.83 (9H, m).

[0113] (4) A colorless oily substance, bis(2-butyloctyl)11-(2-(diethylamino)ethyl)-5,17-dihexyl-7,15-dioxo-6,8,14,16-tetraoxa-11-azahenicosanedioate, was obtained in the same manner as in Example 1, except that 2-butyloctyl 5-(((4-nitrophenoxy)carbonyl)oxy)undecanoate was used instead of 2-hexyldecyl 10-(((4-nitrophenoxy)carbonyl)oxy)hexadecanoate. 1 H-NMR(CDCl3)δ: 4.72-4.65 (2H, m), 4.22-4.09 (4H, m), 3.97 (4H, d, J=6.0Hz), 2.84 (4H, t, J=6.4Hz), 2.70-2.65 (2H, m), 2.55-2.48 (6H, MS m / z(M+H):999. clogP:20.8544

[0114] [Example 9] A colorless oily substance, bis(2-hexyldecyl)11-(2-(diethylamino)ethyl)-5,17-dihexyl-7,15-dioxo-6,8,14,16-tetraoxa-11-azahenicosanedioate, was obtained in the same manner as in Example 1, except that 2-hexyl-1-decanol was used instead of 2-butyl-1-octanol in Example 8. 1H-NMR(CDCl3)δ: 4.72-4.65 (2H, m), 4.22-4.09 (4H, m), 3.97 (4H, d, J=6.0Hz), 2.84 (4H, t, J=6.4Hz), 2.70-2.65 (2H, m), 2.55-2.48 (6H, MS m / z(M+H):1111. clogP:25.0864

[0115] [Example 10] (1) A colorless oily substance, 2-pentylheptyl 5-hydroxyundecanoate, was obtained in the same manner as in Example 8(1) and Example 8(2), except that 2-pentyl-1-heptanol was used instead of 2-butyl-1-octanol. 1 H-NMR(CDCl3)δ: 3.98 (2H, d, J=6.0Hz), 3.63-3.57 (1H, m), 2.41-2.28 (2H, m), 1.84-1.22 (32H, m), 0.88 (9H, t, J=7.2Hz).

[0116] (2) To a solution of 2-pentylheptyl 5-hydroxyundecanoate (0.50 g) in tetrahydrofuran (5.0 mL) was added 1,1'-carbonyldiimidazole (0.33 g), and the mixture was stirred at room temperature for 30 hours. Water (10 mL) and hexane (20 mL) were added to the reaction mixture, and the organic layer was separated and washed with water and saturated aqueous sodium chloride solution. After drying over anhydrous sodium sulfate, the solvent was evaporated under reduced pressure to give 1-oxo-1-((2-pentylheptyl)oxy)undecan-5-yl 1H-imidazole-1-carboxylate (0.64 g) as a pale yellow oil. 1H-NMR(CDCl3)δ: 8.14-8.12 (1H, m), 7.43-7.41 (1H, m), 7.08-7.06 (1H, m), 5.11-5.04 (1H, m), 3.97 (2H, d, J=5.6Hz), 2.38-2.32 (2H, m), 1.80-1.54 (7H, m), 1.40-1.22 (24H, m), 0.91-0.85 (9H, m).

[0117] (3) To a mixture of 1-oxo-1-((2-pentylheptyl)oxy)undecan-5-yl 1H-imidazole-1-carboxylate (0.50 g), 2,2'-((2-(diethylamino)ethyl)azanediyl)bis(ethan-1-ol) (0.11 g), and acetonitrile (2.5 mL), potassium carbonate (0.33 g) was added and stirred at 80°C for 2 hours. The reaction mixture was cooled to room temperature, and then ethyl acetate (3 mL), hexane (3 mL), water (3 mL), and methanol (1 mL) were added, and the organic layer was separated. The mixture was washed with water and a saturated aqueous solution of sodium chloride, dried over anhydrous sodium sulfate, and then the solvent was evaporated under reduced pressure. The obtained residue was purified by silica gel column chromatography (methanol-ethyl acetate-hexane) to give bis(2-pentylheptyl)11-(2-(diethylamino)ethyl)-5,17-dihexyl-7,15-dioxo-6,8,14,16-tetraoxa-11-azahenicosanedioate (0.24 g) as a colorless oil. 1 H-NMR(CDCl3)δ: 4.72-4.65 (2H, m), 4.22-4.09 (4H, m), 3.97 (4H, d, J=6.0Hz), 2.84 (4H, t, J=6.4Hz), 2.70-2.65 (2H, m), 2.55-2.48 (6H, MS m / z(M+H):999. clogP:20.8544

[0118] [Example 11] (1) A colorless oily substance, 2,2'-((2-(dimethylamino)ethyl)azanediyl)bis(ethan-1-ol), was obtained in the same manner as in Example 1(6), except that 2,2'-((2-(dimethylamino)ethyl)azanediyl)bis(ethan-1-ol) hydrochloride was used instead of 2-bromo-N,N-diethylethan-1-amine hydrobromide. 1 H-NMR(CDCl3)δ: 3.57 (4H, t, J=5.2Hz), 2.71 (4H, t, J=5.2Hz), 2.64 (2H, t, J=5.2Hz), 2.41 (2H, t, J=5.2Hz), 2.61 (6H, s). MS m / z(M+H): 177.

[0119] (2) A colorless oily substance, bis(2-pentylheptyl)11-(2-(dimethylamino)ethyl)-5,17-dihexyl-7,15-dioxo-6,8,14,16-tetraoxa-11-azahenicosanedioate, was obtained in the same manner as in Example 10, except that 2,2'-((2-(dimethylamino)ethyl)azanediyl)bis(ethan-1-ol) was used instead of 2,2'-((2-(diethylamino)ethyl)azanediyl)bis(ethan-1-ol). 1 H-NMR(CDCl3)δ: 4.72-4.65 (2H, m), 4.22-4.09 (4H, m), 3.97 (4H, d, J=6.0Hz), 2.85 (4H, t, J=6.4Hz), 2.77-2.65 (2H, m), 2.46-2.19 (12H, m), 1.73-1.50 (14H, m), 1.33-1.20 (48H, m), 0.91-0.84 (18H, m). MS m / z(M+H):970. clogP:19.7964

[0120] [Example 12] (1) A colorless oily substance, 2,2′-((2-(dimethylamino)ethyl)azanediyl)bis(ethan-1-ol), was obtained in the same manner as in Example 1(6), except that N-(2-bromoethyl)-N-propylpropan-1-amine hydrobromide was used instead of 2-bromo-N,N-diethylethan-1-amine hydrobromide in Example 1(6). 1 H-NMR(CDCl3)δ: 3.58 (4H, t, J=5.4Hz), 2.70 (4H, t, J=5.4Hz), 2.67-2.62 (2H, m), 2.54-2.48 (2H, m), 2.45-2.38 (4H, m), 1.57-1.43 (4H, m), 0.88 (6H, t, J=7.2Hz). MS m / z(M+H):233.

[0121] (2) A colorless oily substance, bis(2-pentylheptyl)11-(2-(dipropylamino)ethyl)-5,17-dihexyl-7,15-dioxo-6,8,14,16-tetraoxa-11-azahenicosanedioate, was obtained in the same manner as in Example 10, except that 2,2'-((2-(dimethylamino)ethyl)azanediyl)bis(ethan-1-ol) was used instead of 2,2'-((2-(diethylamino)ethyl)azanediyl)bis(ethan-1-ol) in Example 10. 1 H-NMR(CDCl3)δ: 4.72-4.65 (2H, m), 4.22-4.09 (4H, m), 3.97 (4H, d, J=6.0Hz), 2.83 (4H, t, J=6.4Hz), 2.69-2.62 (2H, m), 2.52-2.47 (2H, MS m / z(M+H):1027. clogP:21.9124

[0122] [Example 13] A colorless oily substance, bis(2-pentylheptyl)11-(3-(diethylamino)propyl)-5,17-dihexyl-7,15-dioxo-6,8,14,16-tetraoxa-11-azahenicosanedioate, was obtained in the same manner as in Example 10, except that 2,2'-((3-(diethylamino)propyl)azanediyl)bis(ethan-1-ol) was used instead of 2,2'-((2-(diethylamino)ethyl)azanediyl)bis(ethan-1-ol) in Example 10. 1 H-NMR(CDCl3)δ: 4.72-4.65 (2H, m), 4.22-4.09 (4H, m), 3.97 (4H, d, J=6.0Hz), 2.80 (4H, t, J=6.4Hz), 2.61-2.45 (6H, m), 2.44-2.37 (2H, MS m / z(M+H):1013. clogP:21.099

[0123] [Example 14] (1) Potassium carbonate (1.5 g) was added to a mixture of N1,N1-diethylbutane-1,4-diamine (0.51 g), 2-bromoethan-1-ol (1.1 g), and acetonitrile (5 mL), and the mixture was stirred under reflux for 2 hours. After the reaction mixture was cooled to room temperature, insoluble materials were removed by filtration, and the solvent was evaporated under reduced pressure. The resulting residue was purified by silica gel column chromatography (ethyl acetate-hexane, NH silica gel) to give 2,2'-((4-(diethylamino)butyl)azanediyl)bis(ethan-1-ol) (0.18 g) as a pale yellow oil. 1 H-NMR(CDCl3)δ: 3.62 (4H, t, J=5.2Hz), 2.65 (4H, t, J=5.2Hz), 2.57-2.49 (6H, m), 2.44-2.39 (2H, m), 1.53-1.43 (4H, m), 1.02 (6H, t, J=7.2Hz). MS m / z(M+H):233.

[0124] (2) A colorless oily substance, bis(2-pentylheptyl)11-(4-(diethylamino)butyl)-5,17-dihexyl-7,15-dioxo-6,8,14,16-tetraoxa-11-azahenicosanedioate, was obtained in the same manner as in Example 10, except that 2,2'-((4-(diethylamino)butyl)azanediyl)bis(ethan-1-ol) was used instead of 2,2'-((2-(diethylamino)ethyl)azanediyl)bis(ethan-1-ol) in Example 10. 1 H-NMR(CDCl3)δ: 4.72-4.65 (2H, m), 4.22-4.09 (4H, m), 3.97 (4H, d, J=6.0Hz), 2.80 (4H, t, J=6.4Hz), 2.58-2.47 (6H, m), 2.44-2.37 (2H, MS m / z(M+H):1027. clogP:20.68

[0125] [Example 15] (1) A pale yellow oily substance, 3,3'-((2-(diethylamino)ethyl)azanediyl)bis(propan-1-ol), was obtained in the same manner as in Example 1(6), except that 3,3'-azanediylbis(propan-1-ol was used instead of 2,2'-azanediylbis(ethan-1-ol). 1 H-NMR(CDCl3)δ: 3.74 (4H, t, J=5.2Hz), 2.61-2.51 (12H, m), 1.74-1.66 (4H, m), 1.04 (6H, t, J=7.2Hz). MS m / z(M+H):233.

[0126] (2) A colorless oily substance, bis(2-pentylheptyl)12-(2-(diethylamino)ethyl)-5,19-dihexyl-7,17-dioxo-6,8,16,18-tetraoxa-12-azatricosandioate, was obtained in the same manner as in Example 10, except that 3,3'-((2-(diethylamino)ethyl)azanediyl)bis(propan-1-ol) was used instead of 2,2'-((2-(diethylamino)ethyl)azanediyl)bis(ethan-1-ol). 1 H-NMR(CDCl3)δ: 4.72-4.65 (2H, m), 4.22-4.09 (4H, m), 3.97 (4H, d, J=6.0Hz), 2.56-2.46 (12H, m), 2.32 (4H, t, J=7.2Hz), 1.83-1.75 (4H, MS m / z(M+H):1027. clogP:21.4444

[0127] [Example 16] (1) A pale yellow oily substance, 4,4'-((2-(diethylamino)ethyl)azanediyl)bis(butan-1-ol), was obtained in the same manner as in Example 1(6), except that 4,4'-azanediylbis(butan-1-ol) was used instead of 2,2'-azanediylbis(ethan-1-ol). 1 H-NMR(CDCl3)δ:3.62-3.58 (4H, m), 2.59-2.46 (12H, m), 1.66-1.59 (8H, m), 1.03 (6H, t, J=7.2Hz). MS m / z(M+H):261.

[0128] (2) A colorless oily substance, bis(2-pentylheptyl)13-(2-(diethylamino)ethyl)-5,21-dihexyl-7,19-dioxo-6,8,18,20-tetraoxa-13-azapentacosanedioate, was obtained in the same manner as in Example 10, except that 4,4'-((2-(diethylamino)ethyl)azanediyl)bis(butan-1-ol) was used instead of 2,2'-((2-(diethylamino)ethyl)azanediyl)bis(ethan-1-ol). 1 H-NMR(CDCl3)δ: 4.72-4.65 (2H, m), 4.22-4.09 (4H, m), 3.97 (4H, d, J=6.0Hz), 2.60-2.41 (12H, m), 2.32 (4H, t, J=7.2Hz), 1.75-1.45 (22H, m), 1.35-1.21 (48H, m), 1.03 (6H, t, J=6.8Hz), 0.91-0.84 (18H, m). MS m / z(M+H):1055. clogP:20.9424

[0129] [Example 17] (1) Ethyl 4-oxodecanoate was obtained as a colorless oil in the same manner as in Example 7(2), except that a 1.0 mol / L hexylmagnesium bromide-tetrahydrofuran solution was used instead of the 1.0 mol / L pentylmagnesium bromide-tetrahydrofuran solution, and ethyl 4-chloro-4-oxobutanoate was used instead of methyl 10-chloro-10-oxodecanoate. 1 H-NMR(CDCl3)δ: 4.13 (2H, q, J=6.8Hz), 2.72 (2H, t, J=7.2Hz), 2.57 (2H, t, J=7.2Hz), 2.45 (2H, t, J=7.2Hz), 1.62-1.51 (2H, m), 1.33-1.23 (9H, m), 0.88 (3H, t, J=6.8Hz).

[0130] (2) To a mixture of ethyl 4-oxodecanoate (2.0 g), tetrahydrofuran (4.0 mL), and ethanol (2.0 mL), 7.0 mol / L aqueous potassium hydroxide solution was added and stirred at 40°C for 45 minutes. After the reaction mixture was cooled to room temperature, 20% aqueous potassium hydrogen sulfate solution (15 mL), ethyl acetate (10 mL), and water (10 mL) were added. The organic layer was separated and dried over anhydrous sodium sulfate, and the solvent was evaporated under reduced pressure. Hexane (6 mL) was added to the resulting residue under ice-cooling, and the solid was collected by filtration, washed with ice-cooled hexane, and then dried under reduced pressure to give 4-oxodecanoic acid (1.5 g) as a white solid. 1 H-NMR(CDCl3)δ: 2.75-2.68 (2H, m), 2.66-2.61 (2H, m), 2.45 (2H, t, J=7.2Hz), 1.63-1.54 (2H, m), 1.34-1.23 (6H, m), 0.88 (3H, t, J=6.8Hz).

[0131] (3) A colorless oily product, 2-pentylheptyl 4-hydroxydecanoate, was obtained in the same manner as in Example 8(2), except that 4-oxoundecanoic acid was used instead of 5-oxoundecanoic acid and 2-pentylpentan-1-ol was used instead of 2-butyloctan-1-ol. 1 H-NMR(CDCl3)δ: 3.97 (2H, d, J=6.0Hz), 3.65-3.58 (1H, m), 2.49-2.43 (2H, m), 1.89-1.56 (7H, m), 1.49-1.23 (22H, m), 0.89 (9H, t, J=6.8Hz).

[0132] (4) A colorless oily substance, bis(2-pentylheptyl)10-(2-(diethylamino)ethyl)-4,16-dihexyl-6,14-dioxo-5,7,13,15-tetraoxa-10-azanonadecanedioate, was obtained in the same manner as in Example 10(2) and Example 10(3), except that 2-pentylheptyl 4-hydroxydecanoate was used instead of 2-pentylheptyl 5-hydroxyundecanoate in Example 10(2). 1 H-NMR(CDCl3)δ: 4.75-4.68 (2H, m), 4.23-4.09 (4H, m), 3.97 (4H, d, J=6.0Hz), 2.84 (4H, t, J=6.4Hz), 2.71-2.64 (2H, m), 2.55-2.48 (6H, MS m / z(M+H):970.clogP:20.9884

[0133] [Example 18] A colorless oily substance, bis(2-pentylheptyl)12-(2-(diethylamino)ethyl)-6,18-dihexyl-8,16-dioxo-7,9,15,17-tetraoxa-12-azatricosane dioate, was obtained in the same manner as in Example 17, except that methyl 6-chloro-6-oxohexanoate was used instead of ethyl 4-chloro-4-oxobutanoate in Example 17. 1H-NMR(CDCl3)δ: 4.71-4.63 (2H, m), 4.22-4.09 (4H, m), 3.97 (4H, d, J=6.0Hz), 2.84 (4H, t, J=6.4Hz), 2.70-2.65 (2H, m), 2.55-2.48 (6H, MS m / z(M+H):1027. clogP:21.9124

[0134] [Example 19] (1) A colorless oily product, 2-pentylheptyl 5-hydroxyhexanoate, was obtained in the same manner as in Example 8(2), except that 5-oxohexanoic acid was used instead of 5-oxoundecanoic acid. 1 H-NMR(CDCl3)δ: 3.98 (2H, d, J=6.0Hz), 3.85-3.76 (1H, m), 2.41-2.86 (2H, m), 1.81-1.54 (3H, m), 1.51-1.43 (3H, m), 1.31-1.21 (16H, m), 1.20 (3H, d, J=6.0Hz), 0.89 (6H, t, J=6.8Hz)

[0135] (2) A colorless oily substance, bis(2-pentylheptyl)11-(2-(diethylamino)ethyl)-5,17-dimethyl-7,15-dioxo-6,8,14,16-tetraoxa-11-azahenicosanedioate, was obtained in the same manner as in Example 10(2) and Example 10(3), except that 2-pentylheptyl 5-hydroxyhexanoate was used instead of 2-pentylheptyl 5-hydroxyundecanoate. 1H-NMR(CDCl3)δ: 4.78-4.71 (2H, m), 4.21-4.10 (4H, m), 3.97 (4H, d, J=5.6Hz), 2.84 (4H, t, J=6.4Hz), 2.69-2.65 (2H, m), 2.55-2.49 (6H, MS m / z(M+H):858. clogP:15.5644

[0136] [Example 20] A colorless oily substance, bis(2-pentylheptyl)11-(2-(diethylamino)ethyl)-5,17-diethyl-7,15-dioxo-6,8,14,16-tetraoxa-11-azahenicosanedioate, was obtained in the same manner as in Example 10, except that a 1.0 mol / L ethylmagnesium bromide-tetrahydrofuran solution was used instead of the 1.0 mol / L hexylmagnesium bromide-tetrahydrofuran solution used in Example 10. 1 H-NMR(CDCl3)δ: 4.67-4.61 (2H, m), 4.22-4.10 (4H, m), 3.97 (4H, d, J=5.6Hz), 2.84 (4H, t, J=6.4Hz), 2.69-2.65 (2H, m), 2.55-2.49 (6H, MS m / z(M+H):886. clogP:16.6224

[0137] [Example 21] A colorless oily substance, bis(2-pentylheptyl)11-(2-(diethylamino)ethyl)-7,15-dioxo-5,17-dipropyl-6,8,14,16-tetraoxa-11-azahenicosanedioate, was obtained in the same manner as in Example 10, except that a 2.0 mol / L propylmagnesium bromide-tetrahydrofuran solution was used instead of the 1.0 mol / L hexylmagnesium bromide-tetrahydrofuran solution used in Example 10. 1 H-NMR(CDCl3)δ: 4.74-4.67 (2H, m), 4.22-4.10 (4H, m), 3.97 (4H, d, J=5.6Hz), 2.83 (4H, t, J=6.4Hz), 2.69-2.65 (2H, m), 2.55-2.49 (6H, MS m / z(M+H):914. clogP:17.6804

[0138] [Example 22] A colorless oily substance, bis(2-pentylheptyl)5,17-dibutyl-11-(2-(diethylamino)ethyl)-7,15-dioxo-6,8,14,16-tetraoxa-11-azahenicosanedioate, was obtained in the same manner as in Example 10, except that a 2.0 mol / L butylmagnesium chloride-tetrahydrofuran solution was used instead of the 1.0 mol / L hexylmagnesium bromide-tetrahydrofuran solution used in Example 10. 1H-NMR(CDCl3)δ: 4.72-4.65 (2H, m), 4.22-4.09 (4H, m), 3.97 (4H, d, J=6.0Hz), 2.84 (4H, t, J=6.4Hz), 2.70-2.65 (2H, m), 2.55-2.48 (6H, MS m / z(M+H):942. clogP:18.7384

[0139] [Example 23] A colorless oily substance, bis(2-pentylheptyl)11-(2-(diethylamino)ethyl)-7,15-dioxo-5,17-dipentyl-6,8,14,16-tetraoxa-11-azahenicosanedioate, was obtained in the same manner as in Example 10, except that a 2.0 mol / L butylmagnesium chloride-tetrahydrofuran solution was used instead of the 1.0 mol / L hexylmagnesium bromide-tetrahydrofuran solution used in Example 10. 1 H-NMR(CDCl3)δ: 4.72-4.65 (2H, m), 4.22-4.09 (4H, m), 3.97 (4H, d, J=6.0Hz), 2.84 (4H, t, J=6.4Hz), 2.70-2.65 (2H, m), 2.56-2.48 (6H, MS m / z(M+H):970. clogP:19.7964

[0140] [Example 24] A colorless oily substance, bis(2-pentylheptyl)11-(2-(diethylamino)ethyl)-5,17-diheptyl-7,15-dioxo-6,8,14,16-tetraoxa-11-azahenicosanedioate, was obtained in the same manner as in Example 10, except that a 1.0 mol / L heptylmagnesium bromide-tetrahydrofuran solution was used instead of the 1.0 mol / L hexylmagnesium bromide-tetrahydrofuran solution used in Example 10. 1 H-NMR(CDCl3)δ: 4.72-4.65 (2H, m), 4.22-4.09 (4H, m), 3.97 (4H, d, J=6.0Hz), 2.84 (4H, t, J=6.4Hz), 2.70-2.65 (2H, m), 2.56-2.48 (6H, MS m / z(M+H):1027. clogP:21.9124

[0141] [Example 25] A colorless oily substance, bis(2-pentylheptyl)11-(2-(diethylamino)ethyl)-5,17-dioctyl-7,15-dioxo-6,8,14,16-tetraoxa-11-azahenicosanedioate, was obtained in the same manner as in Example 10, except that a 2.0 mol / L octylmagnesium bromide-diethyl ether solution was used instead of the 1.0 mol / L hexylmagnesium bromide-tetrahydrofuran solution used in Example 10. 1H-NMR(CDCl3)δ: 4.72-4.65 (2H, m), 4.22-4.09 (4H, m), 3.97 (4H, d, J=6.0Hz), 2.84 (4H, t, J=6.4Hz), 2.70-2.65 (2H, m), 2.56-2.48 (6H, MS m / z(M+H):1055. clogP:22.9704

[0142] [Example 26] A colorless oily substance, bis(2-hexyloctyl)5,17-dibutyl-11-(2-(diethylamino)ethyl)-7,15-dioxo-6,8,14,16-tetraoxa-11-azahenicosanedioate, was obtained in the same manner as in Example 22, except that 2-hexyloctan-1-ol was used instead of 2-pentylheptan-1-ol. 1 H-NMR(CDCl3)δ: 4.72-4.65 (2H, m), 4.22-4.09 (4H, m), 3.97 (4H, d, J=6.0Hz), 2.84 (4H, t, J=6.4Hz), 2.70-2.65 (2H, m), 2.55-2.48 (6H, MS m / z(M+H):999. clogP:20.8544

[0143] [Example 27] A colorless oily substance, bis(2-((3r,5r,7r)-adamantan-1-yl)ethyl)11-(2-(diethylamino)ethyl)-5,17-dihexyl-7,15-dioxo-6,8,14,16-tetraoxa-11-azahenicosanedioate, was obtained in the same manner as in Example 10, except that 2-(1-adamantyl)ethanol was used instead of 2-pentylheptan-1-ol. 1 H-NMR(CDCl3)δ: 4.72-4.65 (2H, m), 4.22-4.09 (8H, m), 2.84 (4H, t, J=6.4Hz), 2.70-2.65 (2H, m), 2.55-2.48 (6H, m), 2.30 (4H, t, MS m / z(M+H):986. clogP:19.1704

[0144] [Example 28] A colorless oily substance, bis(2-pentylheptyl)11-(3-(diethylamino)propyl)-7,15-dioxo-5,17-dipropyl-6,8,14,16-tetraoxa-11-azahenicosanedioate, was obtained in the same manner as in Example 21, except that 2,2'-((3-(diethylamino)propyl)azanediyl)bis(ethan-1-ol) was used instead of 2,2'-((2-(diethylamino)ethyl)azanediyl)bis(ethan-1-ol) in Example 21. 1H-NMR(CDCl3)δ: 4.73-4.67 (2H, m), 4.22-4.10 (4H, m), 3.97 (4H, d, J=5.6Hz), 2.80 (4H, t, J=6.4Hz), 2.58-2.47 (6H, m), 2.43-2.39 (2H, MS m / z(M+H):928. clogP:17.925

[0145] [Example 29] (1) Ethyl acrylate (24.0 mL) was added to a mixture of cyclohexane-1,3-dione (16.5 g), potassium carbonate (20.3 g), benzyltriethylammonium chloride (33.5 g), and dimethyl sulfoxide (165 mL), and the mixture was stirred at 60°C for 4 hours. After the reaction mixture was cooled to room temperature, N-acetyl-L-cysteine ​​(14.4 g) was added and the mixture was stirred at room temperature for 1 hour. Ethyl acetate (165 mL) and 20% aqueous potassium hydrogen sulfate solution (330 mL) were added to the reaction mixture, and the organic layer was separated. The mixture was washed twice with 20% aqueous potassium hydrogen sulfate solution (100 mL), then with water (100 mL), dried over anhydrous sodium sulfate, and the solvent was evaporated under reduced pressure. Ethyl acetate (30 mL) and hexane (60 mL) were added to the resulting residue, and the mixture was stirred at room temperature for 20 minutes. The solid matter was collected by filtration, washed with a 33% ethyl acetate-hexane solution, and then dried under reduced pressure to give ethyl 3-(2,6-dioxocyclohexyl)propanoate (20.2 g) as a pale yellow solid. 1 H-NMR(CDCl3)δ: 9.57 (1H, s), 4.18 (2H, q, J=7.2Hz), 2.58-2.44 (6H, m), 2.32 (2H, t, J=6.8Hz), 1.95-1.87 (2H, m), 1.27 (3H, t, J=6.8Hz). MS m / z(M+H):213.

[0146] To ethyl 3-(2,6-dioxocyclohexyl)propanoate (20.2 g), 10% aqueous hydrochloric acid (200 mL) was added and the mixture was stirred at 100°C for 4 hours. After the reaction mixture was cooled to room temperature, ethyl acetate (200 mL) was added and the organic layer was separated. The aqueous layer was extracted three times with ethyl acetate (50 mL). The organic layer and the extract were combined and dried over anhydrous sodium sulfate. The solvent was then evaporated under reduced pressure. Ethyl acetate (20 mL) and hexane (40 mL) were added to the resulting residue, and the solid was collected by filtration, washed with a 33% ethyl acetate-hexane solution, and dried under reduced pressure to give 5-oxononanedioic acid (9.1 g) as a pale yellow solid. 1 H-NMR(CDCl3)δ: 2.43 (4H, t, J=7.2Hz), 2.19 (4H, t, J=7.2Hz), 1.66 (4H, quin, J=7.2Hz). MS m / z(MH):201.

[0147] (2) To a mixture of 5-oxononanedioic acid (0.500 g), heptan-1-ol (0.517 g), and toluene (1.0 mL), p-toluenesulfonic acid (0.023 g) was added and stirred at 100° C. for 2 hours. The reaction mixture was cooled to room temperature and then purified by silica gel column chromatography (ethyl acetate-hexane) to obtain diheptyl 5-oxononanedioate (0.746 g) as a colorless oil. 1 H-NMR(CDCl3)δ: 4.06 (4H, d, J=6.8Hz), 2.47 (4H, t, J=7.2Hz), 2.32 (4H, t, J=7.2Hz), 1.93-1.85 (4H, m), 1.66-1.56 (4H, m), 1.36-1.24 (16H, m), 0.89 (6H, t, J=6.8Hz).

[0148] To a mixture of diheptyl 5-oxononanedioate (0.746 g), toluene (3.0 mL), and methanol (3.0 mL), sodium borohydride (0.085 g) was added under ice-cooling, and the mixture was stirred at the same temperature for 1 hour. Water (3.0 mL), 1.0 mol / L aqueous hydrochloric acid (3.0 mL), and ethyl acetate were added to the reaction mixture under ice-cooling, and the organic layer was separated. After washing with saturated aqueous sodium chloride and drying over anhydrous sodium sulfate, the solvent was evaporated under reduced pressure. The resulting residue was purified by silica gel column chromatography (ethyl acetate-hexane) to yield diheptyl 5-hydroxynonanedioate (0.656 g) as a colorless oil. 1 H-NMR(CDCl3)δ: 4.06 (4H, d, J=6.8Hz), 3.65-3.56 (1H, m), 2.39-2.27 (4H, m), 1.83-1.39 (12H, m), 1.36-1.23 (16H, m), 0.88 (6H, t, J=6.8Hz).

[0149] To a solution of diheptyl 5-hydroxynonanedioate (0.656 g) in tetrahydrofuran (4.0 mL) was added 1,1'-carbonyldiimidazole (0.398 g) and the mixture was stirred at room temperature for 3 hours. Water (4 mL) and hexane (4 mL) were added to the reaction mixture, and the organic layer was separated and washed with water and a saturated aqueous solution of sodium chloride. After drying over anhydrous sodium sulfate, the solvent was evaporated under reduced pressure to give diheptyl 5-((1H-imidazole-1-carbonyl)oxy)nonanedioate (0.82 g) as a pale yellow oil. 1 H-NMR(CDCl3)δ: 8.14-8.12 (1H, m), 7.43-7.41 (1H, m), 7.08-7.06 (1H, m), 5.12-5.05 (1H, m), 4.06 (4H, d, J=6.4Hz), 2.35 (4H, t, J=6.4Hz), 1.80-1.54 (12H, m), 1.35-1.23 (16H, m), 0.88 (6H, t, J=7.2Hz).

[0150] (3) To a mixture of diheptyl 5-((1H-imidazole-1-carbonyl)oxy)nonanedioate (0.82 g), 2,2'-((2-(diethylamino)ethyl)azanediyl)bis(ethan-1-ol) (0.17 g), and acetonitrile (4.0 mL), potassium carbonate (0.45 g) was added and stirred at 80°C for 2 hours. After the reaction mixture was cooled to room temperature, ethyl acetate (4 mL) and water (4 mL) were added, the organic layer was separated, and the solvent was evaporated under reduced pressure. To the resulting residue, ethyl acetate (4 mL), hexane (4 mL), and water (4 mL) were added, and the organic layer was separated. After drying over anhydrous sodium sulfate, the solvent was evaporated under reduced pressure. The obtained residue was purified by silica gel column chromatography (methanol-ethyl acetate-hexane) to give diheptyl 11-(2-(diethylamino)ethyl)-5,17-bis(4-(heptyloxy)-4-oxobutyl)-7,15-dioxo-6,8,14,16-tetraoxa-11-azahenicosanedioate (0.39 g). 1 H-NMR(CDCl3)δ: 4.74-4.66 (2H, m), 4.15 (4H, t, J=6.0Hz), 4.05 (8H, t, J=6.8Hz), 2.84 (4H, t, J=6.4Hz), 2.70-2.64 (2H, m), 2.55-2.48 (6H, m), 2.34-2.28 (8H, m), 1.72-1.54 (24H, m), 1.36-1.24 (32H, m), 1.01 (6H, t, J=7.2Hz), 0.88 (12H, t, J=6.8Hz). MS m / z(M+H):1059. clogP:16.9164

[0151] [Example 30] A colorless oily substance, bis(2-hexyloctyl)11-(2-(diethylamino)ethyl)-5,17-bis(4-((2-hexyloctyl)oxy)-4-oxobutyl)-7,15-dioxo-6,8,14,16-tetraoxa-11-azahenicosanedioate, was obtained in the same manner as in Example 29, except that 2-hexyloctan-1-ol was used instead of heptan-1-ol. 1H-NMR(CDCl3)δ: 4.75-4.66 (2H, m), 4.15 (4H, t, J=6.0Hz), 3.96 (8H, d, J=5.6Hz), 2.83 (4H, t, J=6.4Hz), 2.70-2.64 (2H, m), 2.55-2.48 (6H, m), 2.34-2.28 (8H, m), 1.73-1.56 (20H, m), 1.36-1.24 (80H, m), 1.01 (6H, t, J=7.2Hz), 0.88 (24H, t, J=6.8Hz). MS m / z(M+H):1451. clogP: 31.2084

[0152] [Example 31] A colorless oily substance, dipentyl 11-(2-(diethylamino)ethyl)-7,15-dioxo-5,17-bis(4-oxo-4-(pentyloxy)butyl)-6,8,14,16-tetraoxa-11-azahenicosanedioate, was obtained in the same manner as in Example 29, except that pentan-1-ol was used instead of heptan-1-ol. 1 H-NMR(CDCl3)δ: 4.74-4.66 (2H, m), 4.15 (4H, t, J=6.0Hz), 4.05 (8H, t, J=6.8Hz), 2.84 (4H, t, J=6.4Hz), 2.70-2.64 (2H, m), 2.55-2.48 (6H, m), 2.34-2.28 (8H, m), 1.72-1.54 (24H, m), 1.36-1.24 (16H, m), 1.01 (6H, t, J=7.2Hz), 0.88 (12H, t, J=6.8Hz). MS m / z(M+H):946. clogP:12.6844

[0153] [Example 32] A colorless oily substance, dihexyl 11-(2-(diethylamino)ethyl)-5,17-bis(4-(hexyloxy)-4-oxobutyl)-7,15-dioxo-6,8,14,16-tetraoxa-11-azahenicosanedioate, was obtained in the same manner as in Example 29, except that hexane-1-ol was used instead of heptan-1-ol.1 H-NMR(CDCl3)δ: 4.74-4.66 (2H, m), 4.15 (4H, t, J=6.0Hz), 4.05 (8H, t, J=6.8Hz), 2.84 (4H, t, J=6.4Hz), 2.70-2.64 (2H, m), 2.55-2.48 (6H, m), 2.34-2.28 (8H, m), 1.72-1.54 (24H, m), 1.36-1.24 (24H, m), 1.01 (6H, t, J=7.2Hz), 0.88 (12H, t, J=6.8Hz). MS m / z(M+H):1002. clogP:14.8004

[0154] [Example 33] A colorless oily substance, dioctyl 11-(2-(diethylamino)ethyl)-5,17-bis(4-(octyloxy)-4-oxobutyl)-7,15-dioxo-6,8,14,16-tetraoxa-11-azahenicosanedioate, was obtained in the same manner as in Example 29, except that octan-1-ol was used instead of heptan-1-ol. 1 H-NMR(CDCl3)δ: 4.74-4.66 (2H, m), 4.15 (4H, t, J=6.0Hz), 4.05 (8H, t, J=6.8Hz), 2.84 (4H, t, J=6.4Hz), 2.70-2.64 (2H, m), 2.55-2.48 (6H, m), 2.34-2.28 (8H, m), 1.72-1.54 (24H, m), 1.36-1.24 (40H, m), 1.01 (6H, t, J=7.2Hz), 0.88 (12H, t, J=6.8Hz). MS m / z(M+H):1115. clogP:19.0324

[0155] [Example 34] A colorless oily substance, dinonyl 11-(2-(diethylamino)ethyl)-5,17-bis(4-(nonyloxy)-4-oxobutyl)-7,15-dioxo-6,8,14,16-tetraoxa-11-azahenicosanedioate, was obtained in the same manner as in Example 29, except that nonan-1-ol was used instead of heptan-1-ol. 1 H-NMR(CDCl3)δ: 4.74-4.66 (2H, m), 4.15 (4H, t, J=6.0Hz), 4.05 (8H, t, J=6.8Hz), 2.84 (4H, t, J=6.4Hz), 2.70-2.64 (2H, m), 2.55-2.48 (6H, m), 2.34-2.28 (8H, m), 1.72-1.54 (24H, m), 1.36-1.24 (48H, m), 1.01 (6H, t, J=7.2Hz), 0.88 (12H, t, J=6.8Hz). MS m / z(M+H):1171. clogP:21.1484

[0156] [Example 35] A colorless oily substance, didecyl 5,17-bis(4-(decyloxy)-4-oxobutyl)-11-(2-(diethylamino)ethyl)-7,15-dioxo-6,8,14,16-tetraoxa-11-azahenicosanedioate, was obtained in the same manner as in Example 29, except that decan-1-ol was used instead of heptan-1-ol. 1 H-NMR(CDCl3)δ: 4.74-4.66 (2H, m), 4.15 (4H, t, J=6.0Hz), 4.05 (8H, t, J=6.8Hz), 2.84 (4H, t, J=6.4Hz), 2.70-2.64 (2H, m), 2.55-2.48 (6H, m), 2.34-2.28 (8H, m), 1.72-1.54 (24H, m), 1.36-1.24 (56H, m), 1.01 (6H, t, J=7.2Hz), 0.88 (12H, t, J=6.8Hz). MS m / z(M+H):1227. clogP:23.2644

[0157] [Example 36] (1) To a mixture of dihexyl 5-((1H-imidazole-1-carbonyl)oxy)nonanedioate (0.691 g), 2,2'-((2-(diethylamino)ethyl)azanediyl)bis(ethan-1-ol) (0.234 g), and acetonitrile (3.0 mL), potassium carbonate (0.339 g) was added and the mixture was stirred at 80°C for 1 hour and 30 minutes. After the reaction mixture was cooled to room temperature, ethyl acetate (4.0 mL) and water (4.0 mL) were added, and the organic layer was separated and dried over anhydrous sodium sulfate. The solvent was then evaporated under reduced pressure. The resulting residue was purified by silica gel column chromatography (methanol-ethyl acetate-hexane) to give dihexyl 5-(((2-((2-(diethylamino)ethyl)(2-hydroxyethyl)amino)ethoxy)carbonyl)oxy)nonanedioate (0.212 g). 1 H-NMR(CDCl3)δ: 4.73-4.66 (1H, m), 4.20 (2H, t, J=6.0Hz), 4.06 (4H, t, J=6.8Hz), 3.54 (2H, t, J=5.2Hz), 2.88 (2H, t, J=6.2Hz), 2.71-2.66 (4H, m), 2.58-2.47 (6H, m), 2.33-2.29 (4H, m), 1.74-1.55 (13H, m), 1.36-1.27 (12H, m), 1.03 (6H, t, J=7.0Hz), 0.91-0.87 (6H, m). MS m / z(M+H):604.

[0158] (2) To a mixture of dihexyl 5-(((2-((2-(diethylamino)ethyl)(2-hydroxyethyl)amino)ethoxy)carbonyl)oxy)nonanedioate (0.106 g), dipentyl 5-((1H-imidazole-1-carbonyl)oxy)nonanedioate (0.093 g), and acetonitrile (1.0 mL), potassium carbonate (0.830 g) was added and stirred at 80°C for 2 hours. The reaction mixture was cooled to room temperature, and then ethyl acetate (4 mL) and water (4 mL) were added. The organic layer was separated and dried over anhydrous sodium sulfate, and the solvent was evaporated under reduced pressure. The resulting residue was purified by silica gel column chromatography (methanol-ethyl acetate-hexane) to give 1-hexyl 21-pentyl 11-(2-(diethylamino)ethyl)-5-(4-(hexyloxy)-4-oxobutyl)-7,15-dioxo-17-(4-oxo-4-(pentyloxy)butyl)-6,8,14,16-tetraoxa-11-azahenicosanedioate (0.110 g). 1 H-NMR(CDCl3)δ: 4.73-4.66 (2H, m), 4.15 (4H, t, J=6.8Hz), 4.05 (8H, t, J=6.6Hz), 2.83 (4H, t, J=6.4Hz), 2.69-2.65 (2H, m), 2.54-2.49 (6H, m), 2.33-2.29 (8H, m), 1.74-1.55 (24H, m), 1.38-1.25 (20H, m), 1.01 (6H, t, J=7.2Hz), 0.93-0.87 (12H, m). MS m / z(M+H):974. clogP:13.7424

[0159] [Example 37] A colorless oily substance, 1-heptyl 21-hexyl 11-(2-(diethylamino)ethyl)-5-(4-(heptyloxy)-4-oxobutyl)-17-(4-(hexyloxy)-4-oxobutyl)-7,15-dioxo-6,8,14,16-tetraoxa-11-azahenicosanedioate, was obtained in the same manner as in Example 36, except that diheptyl 5-((1H-imidazole-1-carbonyl)oxy)nonanedioate was used instead of dipentyl 5-((1H-imidazole-1-carbonyl)oxy)nonanedioate in Example 36. 1 H-NMR(CDCl3)δ: 4.73-4.66 (2H, m), 4.16 (4H, t, J=6.8Hz), 4.05 (8H, t, J=6.6Hz), 2.83 (4H, t, J=6.0Hz), 2.70-2.66 (2H, m), 2.55-2.49 (6H, m), 2.33-2.29 (8H, m), 1.73-1.56 (24H, m), 1.38-1.23 (28H, m), 1.01 (6H, t, J=7.2Hz), 0.91-0.86 (12H, m). MS m / z(M+H):1030. clogP:15.8584

[0160] [Example 38] A colorless oily substance, diheptyl 11-(3-(diethylamino)propyl)-5,17-bis(4-(heptyloxy)-4-oxobutyl)-7,15-dioxo-6,8,14,16-tetraoxa-11-azahenicosanedioate, was obtained in the same manner as in Example 29, except that 2,2'-((3-(diethylamino)propyl)azanediyl)bis(ethan-1-ol) was used instead of 2,2'-((2-(diethylamino)ethyl)azanediyl)bis(ethan-1-ol) in Example 29. 1H-NMR(CDCl3)δ: 4.73-4.66 (2H, m), 4.15 (4H, t, J=6.8Hz), 4.05 (8H, t, J=6.6Hz), 2.80 (4H, t, J=6.4Hz), 2.58-2.47 (6H, m), 2.42-2.39 (2H, m), 2.33-2.29 (8H, m), 1.74-1.57 (26H, m), 1.35-1.21 (32H, m), 1.00 (6H, t, J=7.2Hz), 0.90-0.86 (12H, m). MS m / z(M+H):1073. clogP:17.161

[0161] [Example 39] (1) To a mixture of diethyl 4-oxoheptanedioate (5.0 g), tetrahydrofuran (10 mL), and ethanol (10 mL) was added 20% aqueous potassium hydroxide solution (14 g), and the mixture was stirred at room temperature for 30 minutes. To the reaction mixture were added 30% aqueous hydrochloric acid solution (10 mL) and ethyl acetate (10 mL), and the organic layer was separated and dried over anhydrous sodium sulfate. The solvent was then evaporated under reduced pressure to give 4-oxoheptanedioic acid (4.3 g) as a pale yellow solid. 1 H-NMR(CDCl3)δ: 2.82-2.74 (4H, m), 2.68-2.58 (4H, m). MS m / z(MH):173.

[0162] (2) A colorless oily substance, diheptyl 10-(2-(diethylamino)ethyl)-4,16-bis(3-(heptyloxy)-3-oxopropyl)-6,14-dioxo-5,7,13,15-tetraoxa-10-azanonadecanedioate, was obtained in the same manner as in Example 29(2) and Example 29(3), except that 4-oxoheptanedioic acid was used instead of 5-oxononanedioic acid. 1H-NMR(CDCl3)δ: 4.80-4.73 (2H, m), 4.16 (4H, t, J=6.0Hz), 4.06 (8H, t, J=6.8Hz), 2.84 (4H, t, J=6.4Hz), 2.70-2.64 (2H, m), 2.55-2.48 MS m / z(M+H):1002.clogP:17.1844

[0163] [Example 40] (1) Potassium carbonate (9.4 g) was added to a mixture of octane-1-thiol (5.0 g), 2-bromoethan-1-ol (4.7 g), and acetonitrile (25 mL), and the mixture was stirred at 60°C for 4 hours. After the reaction mixture was cooled to room temperature, water (25 mL) and hexane (25 mL) were added, and the organic layer was separated and washed with saturated aqueous sodium chloride. After drying over anhydrous sodium sulfate, the solvent was evaporated under reduced pressure. The resulting residue was purified by silica gel column chromatography (ethyl acetate-hexane) to give 2-(octylthio)ethan-1-ol (6.1 g). 1 H-NMR(CDCl3)δ: 3.72 (2H, q, J=6.0Hz), 2.73 (2H, t, J=6.0Hz), 2.52 (2H, t, J=7.2Hz), 2.23 (1H, t, J=6.0Hz), 1.63-1.54 (2H, m), 1.43-1.21 (10H, m), 0.88 (3H, t, J=6.8Hz).

[0164] (2) A colorless oily substance, bis(2-(octylthio)ethyl)11-(2-(diethylamino)ethyl)-5,17-bis(4-(2-(octylthio)ethoxy)-4-oxobutyl)-7,15-dioxo-6,8,14,16-tetraoxa-11-azahenicosanedioate, was obtained in the same manner as in Example 29(2) and Example 29(3), except that 2-(octylthio)ethan-1-ol was used instead of heptan-1-ol. 1 H-NMR(CDCl3)δ: 4.74-4.66 (2H, m), 4.21 (8H, t, J=6.8Hz), 4.16 (4H, t, J=6.4Hz), 2.84 (4H, t, J=6.4Hz), 2.75-2.65 (10H, m), 2.57-2.48 (14H, m), 2.36-2.31 (8H, m), 1.72-1.54 (16H, m), 1.42-1.23 (48H, m), 1.01 (6H, t, J=7.2Hz), 0.88 (12H, t, J=6.8Hz). MS m / z(M+H):1354. clogP:21.954

[0165] [Example 41] A colorless oily substance, bis(cyclohexylmethyl)5,17-bis(4-(cyclohexylmethoxy)-4-oxobutyl)-11-(2-(diethylamino)ethyl)-7,15-dioxo-6,8,14,16-tetraoxa-11-azahenicosanediioate, was obtained in the same manner as in Example 29, except that cyclohexylmethanol was used instead of heptan-1-ol. 1 H-NMR(CDCl3)δ: 4.73-4.66 (2H, m), 4.16 (4H, t, J=6.4Hz), 3.87 (8H, d, J=6.8Hz), 2.84 (4H, t, J=6.4Hz), 2.70-2.64 (2H, m), 2.57-2.46 (6H, m), 2.34-2.30 (8H, m), 1.76-1.55 (40H, m), 1.30-0.90 (26H, m). MS m / z(M+H):1050. clogP:14.8204

[0166] [Example 42] A colorless oily substance, 1-hexyl 21-octyl 11-(2-(diethylamino)ethyl)-5-(4-(hexyloxy)-4-oxobutyl)-17-(4-(octyloxy)-4-oxobutyl)-7,15-dioxo-6,8,14,16-tetraoxa-11-azahenicosanedioate, was obtained in the same manner as in Example 36(2), except that dioctyl 5-((1H-imidazole-1-carbonyl)oxy)nonanedioate was used instead of dipentyl 5-((1H-imidazole-1-carbonyl)oxy)nonanedioate in Example 36(2). 1 H-NMR(CDCl3)δ: 4.73-4.66 (2H, m), 4.16 (4H, t, J=6.4Hz), 4.07-4.03 (8H, m), 2.84 (4H, t, J=6.4Hz), 2.70-2.66 (2H, m), 2.56-2.50 (6H, MS m / z(M+H):1059. clogP:16.9164

[0167] [Example 43] (1) To a mixture of 2-mercaptoethanol (3.0 g), potassium hydroxide (2.6 g), and ethanol (100 mL), 1-bromohexane (5.2 g) was added and stirred at room temperature for 2 hours, after which the solvent was evaporated under reduced pressure. Ethyl acetate (150 mL) and water (200 mL) were added to the resulting residue, and the organic layer was separated and washed with water (100 mL) and saturated aqueous sodium chloride solution. After drying over anhydrous sodium sulfate, the solvent was evaporated under reduced pressure to give 2-(hexylthio)ethan-1-ol (6.2 g). 1H-NMR(CDCl3)δ: 3.72 (2H, dt, J=6.0, 6.0Hz), 2.73 (2H, t, J=6.0Hz), 2.52 (2H, t, J=7.6Hz), 2.22-2.15 (1H, m), 1.63-1.53 ​​(2H, m), 1.42-1.22 (6H, m), 0.89 (3H, t, J=6.8Hz).

[0168] (2) A colorless oily substance, bis(2-(hexylthio)ethyl)11-(2-(diethylamino)ethyl)-5,17-bis(4-(2-(hexylthio)ethoxy)-4-oxobutyl)-7,15-dioxo-6,8,14,16-tetraoxa-11-azahenicosanedioate, was obtained in the same manner as in Example 29(2) and Example 29(3), except that 2-(hexylthio)ethan-1-ol was used instead of heptan-1-ol. 1 H-NMR(CDCl3)δ: 4.72-4.66 (2H, m), 4.21 (8H, t, J=7.0Hz), 4.16 (4H, t, J=6.2Hz), 2.84 (4H, t, J=6.4Hz), 2.73 (8H, t, J=6.8Hz), 2.69-2.65 (2H, m), 2.57-2.49 (14H, m), 2.36-2.32 (8H, m), 1.75-1.54 (24H, m), 1.42-1.23 (24H, m), 1.01 (6H, t, J=7.2Hz), 0.91-0.87 (12H, m). MS m / z(M+H):1242. clogP:17.722

[0169] [Example 44] (1) To a solution of 8-bromooctan-1-ol (5.0 g) in N-methylpyrrolidone (25 mL), 70% aqueous sodium hydrogen sulfide solution (13.4 g) was added under ice-cooling, and the mixture was stirred at room temperature for 1 hour. Water (100 mL), ethyl acetate (50 mL), and hexane (50 mL) were added to the reaction mixture, and the organic layer was separated and washed with water (100 mL) and saturated aqueous sodium chloride solution. After drying over anhydrous sodium sulfate, the solvent was evaporated under reduced pressure to give 8-mercaptooctan-1-ol (3.8 g). 1 H-NMR(CDCl3)δ: 3.68-3.61 (2H, m), 2.53 (2H, dt, J=7.6, 7.6Hz), 1.71-1.53 ​​(5H, m), 1.42-1.20 (9H, m).

[0170] To a mixture of 8-mercaptooctan-1-ol (1.9 g), potassium hydroxide (0.72 g), and ethanol (60 mL), methyl p-toluenesulfonate (2.2 g) was added and stirred at room temperature for 1 hour, after which the solvent was evaporated under reduced pressure. Ethyl acetate (150 mL) and water (100 mL) were added to the resulting residue, and the organic layer was separated and washed with water (100 mL) and saturated aqueous sodium chloride solution. After drying over anhydrous sodium sulfate, the solvent was evaporated under reduced pressure to give 8-(methylthio)octan-1-ol (1.3 g). 1 H-NMR(CDCl3)δ: 3.67-3.62 (2H, m), 2.49 (2H, t, J=7.4Hz), 2.10 (3H, s), 1.63-1.53 ​​(4H, m), 1.42-1.20 (9H, m).

[0171] (2) A colorless oily substance, bis(8-(methylthio)octyl)11-(2-(diethylamino)ethyl)-5,17-bis(4-((8-(methylthio)octyl)oxy)-4-oxobutyl)-7,15-dioxo-6,8,14,16-tetraoxa-11-azahenicosanedioate, was obtained in the same manner as in Example 29(2) and Example 29(3), except that 8-(methylthio)octan-1-ol was used instead of heptan-1-ol. 1H-NMR(CDCl3)δ: 4.72-4.66 (2H, m), 4.16 (4H, t, J=6.4Hz), 4.05 (8H, t, J=6.6Hz), 2.84 (4H, t, J=6.4Hz), 2.69-2.65 (2H, m), 2.55-2.46 (14H, m), 2.33-2.29 (8H, m), 2.10 (12H, s), 1.72-1.56 (32H, m), 1.42-1.31 (32H, m), 1.01 (6H, t, J=7.0Hz). MS m / z(M+H):1298. clogP:17.5084

[0172] The structures of the compounds of Comparative Examples 1 and 2 are shown below.

[0173]

[0174] Test Example 1: Preparation of mRNA-encapsulating lipid particles and measurement of reporter protein expression rate in mice <Preparation of EPO mRNA-encapsulating lipid particles> The compounds listed in Table 1, neutral lipids, cholesterol (product name: Cholesterol HP; Nippon Fine Chemical Co., Ltd.), and 1,2-dimyristoyl-rac-glycero-3-(methylpolyoxyethylene 2000) (hereinafter, DMG-PEG2000) (product name: SUNBRIGHT® GM-020; NOF Corporation) were dissolved in ethanol at the molar ratios shown in Table 1 to give a total lipid concentration of 20 mmol / L, to obtain an oil phase.

[0175] The neutral lipids used were 1,2-distearoyl-sn-glycero-3-phosphocholine (product name: COATSOME® MC-8080; NOF Corporation) (hereinafter referred to as DSPC), L-α-dioleoyl phosphatidylethanolamine (hereinafter referred to as DOPE) (product name: COATSOME® ME-8181; NOF Corporation), or 1,2-dioleoyl-sn-glycero-3-phosphocholine (hereinafter referred to as DOPC) (product name: COATSOME® MC-8181; NOF Corporation).

[0176] EPO mRNA (product name: CleanCap EPO mRNA (5 moU); TriLink) was diluted with 50 mmol / L citrate buffer at pH 4 so that the weight ratio of total lipid concentration to mRNA concentration was approximately 16:1 to 64:1 to obtain an aqueous phase. Subsequently, the aqueous and oil phases were mixed using a NanoAssemblr (Precision NanoSystems) so that the volume ratio of aqueous phase:oil phase = 3:1, and the mixture was diluted 1.5-fold with phosphate-buffered saline (PBS) to obtain a dispersion of mRNA-lipid particles. This dispersion was dialyzed against a 10% sucrose aqueous solution using a dialysis cassette (Slide-A-Lyzer G2, MWCO: 10 kD, Thermo Fisher Scientific) to remove the ethanol, thereby obtaining lipid particles encapsulating EPO mRNA.

[0177] <Measurement of particle size> The particle size of the mRNA-encapsulated lipid particles was measured using a zeta potential / particle size measurement system ELS-Z2 (Otsuka Electronics) after diluting the lipid particle dispersion 10-fold with phosphate buffered saline (PBS). The results are shown in Table 1.

[0178] <Evaluation of mRNA Encapsulation Rate> (Quantification of Total mRNA Concentration) 15-30 μL of 3 mol / L aqueous sodium acetate solution and 4.5-9 μL of glycogen were added to 30-60 μL of lipid particles carrying mRNA, followed by the addition of 0.75-1.5 mL of ethanol to dissolve the lipid and precipitate only the mRNA. The mixture was then centrifuged and the supernatant was removed. After air-drying for 15 minutes or more, the mixture was redissolved by adding water, and the total mRNA concentration was quantified by measuring the concentration using a Nanodrop NF1000 (Thermo Fisher Scientific).

[0179] (Quantification of mRNA concentration in the external aqueous phase) Quantitation was performed using the Quant-iT RiboGreen RNA Assay Kit (Thermo Fisher Scientific) according to the protocol. First, the 20x TE buffer included in the kit was diluted with water to make 1x TE buffer. TE stands for Tris / EDTA (ethylenediaminetetraacetic acid). To quantify only the mRNA in the external aqueous phase, the lipid particle dispersion containing the mRNA was diluted 10,000 times with 1x TE buffer. 100 μL of a lipid particle dispersion diluted 10,000-fold was placed in a 96-well plate, and then 100 μL of RiboGreen reagent (contained in the above-mentioned Quanti-iT Ribogreen RNA Assay Kit) diluted 2,000-fold with 1×TE buffer was added to the sample. Fluorescence (excitation wavelength: 485 nm, fluorescence wavelength: 535 nm) was measured using a plate reader Infinit eF200 (TECAN) to quantify the mRNA concentration in the external aqueous phase.

[0180] (Calculation of Encapsulation Rate) Using the quantitative results of the total mRNA concentration and the mRNA concentration in the external aqueous phase obtained in the above steps, the mRNA encapsulation rate of the mRNA lipid particles was calculated according to the following formula. The results are shown in Table 1. mRNA encapsulation rate (%) = (total mRNA concentration - mRNA concentration in the external aqueous phase) ÷ total mRNA concentration × 100

[0181] <Measurement of EPO enzyme activity> The mRNA lipid particle dispersion prepared in the above <Preparation of EPO mRNA-encapsulating lipid particles> was intravenously administered to C57BL / 6J mice at an mRNA dose of 0.1 mg / kg. 20 to 24 hours after administration, blood was collected from the posterior vena cava to obtain plasma. Human EPO enzyme activity was quantified using the obtained plasma using the ab119522 Erythropoietin (EPO) Human Elisa Kit (Abcam). Quantitative values ​​were expressed as relative EPO protein amounts, with Comparative Example 1 set to 1. The results are shown in Table 1.

[0182]

[0183] It was shown that the nucleic acid-lipid composition of the present invention had a higher EPO protein expression rate than the nucleic acid-lipid composition of the comparative example.

[0184] Test Example 2: Preparation of mRNA-encapsulating lipid particles and measurement of reporter protein expression rate in mice <Preparation of FLuc mRNA-encapsulating lipid particles> The compounds listed in Table 2, neutral lipids, cholesterol (product name: Cholesterol HP; Nippon Fine Chemical Co., Ltd.), and 1,2-dimyristoyl-rac-glycero-3-(methylpolyoxyethylene 2000) (hereinafter, DMG-PEG2000) (in the molar ratios shown in Table 2) were dissolved in ethanol to a total lipid concentration of 20 mmol / L to obtain an oil phase.

[0185] The neutral lipid used in Comparative Example 1 was 1,2-distearoyl-sn-glycero-3-phosphocholine (product name: COATSOME® MC-8080; NOF Corporation), and in all other cases, L-α-dioleoylphosphatidylethanolamine (product name: COATSOME® MC-8181; NOF Corporation).

[0186] FLuc mRNA (product name: CleanCap FLuc mRNA (5 moU); TriLink) was diluted with 50 mmol / L citrate buffer at pH 4 so that the weight ratio of total lipid concentration to mRNA concentration was approximately 19:1 to 64:1 to obtain an aqueous phase. Subsequently, the aqueous and oil phases were mixed using a NanoAssemblr (Precision NanoSystems) so that the volume ratio of aqueous phase:oil phase = 3:1, and the mixture was diluted 1.5-fold with phosphate-buffered saline (PBS) to obtain a dispersion of mRNA-lipid particles. This dispersion was dialyzed against a 10% aqueous sucrose solution using a dialysis cassette (Slide-A-Lyzer G2, MWCO: 10 kD, Thermo Fisher Scientific) to remove the ethanol, thereby obtaining FLuc mRNA-encapsulating lipid particles.

[0187] <Measurement of particle size> The particle size of the mRNA-encapsulated lipid particles was measured using a zeta potential / particle size measurement system ELS-Z2 (Otsuka Electronics) after diluting the lipid particle dispersion 10-fold with phosphate buffered saline (PBS). The results are shown in Table 2.

[0188] <Evaluation of mRNA Encapsulation Rate> (Quantification of Total mRNA Concentration) 15-30 μL of 3 mol / L aqueous sodium acetate solution and 4.5-9 μL of glycogen were added to 30-60 μL of lipid particles carrying mRNA, followed by the addition of 0.75-1.5 mL of ethanol to dissolve the lipid and precipitate only the mRNA. The mixture was then centrifuged and the supernatant was removed. After air-drying for 15 minutes or more, the mixture was redissolved by adding water, and the total mRNA concentration was quantified by measuring the concentration using a Nanodrop NF1000 (Thermo Fisher Scientific).

[0189] (Quantification of mRNA concentration in the external aqueous phase) Quantitation was performed using the Quant-iT RiboGreen RNA Assay Kit (Thermo Fisher Scientific) according to the protocol. First, the 20x TE buffer included in the kit was diluted with water to make 1x TE buffer. TE stands for Tris / EDTA (ethylenediaminetetraacetic acid). To quantify only the mRNA in the external aqueous phase, the lipid particle dispersion containing the mRNA was diluted 10,000 times with 1x TE buffer. 100 μL of a lipid particle dispersion diluted 10,000-fold was placed in a 96-well plate, and then 100 μL of RiboGreen reagent (contained in the above-mentioned Quanti-iT Ribogreen RNA Assay Kit) diluted 2,000-fold with 1×TE buffer was added to the sample. Fluorescence (excitation wavelength: 485 nm, fluorescence wavelength: 535 nm) was measured using a plate reader Infinit EF200 (TECAN) to quantify the mRNA concentration in the external aqueous phase.

[0190] (Calculation of Encapsulation Rate) Using the quantitative results of the total mRNA concentration and the mRNA concentration in the external aqueous phase obtained in the above steps, the mRNA encapsulation rate of the mRNA lipid particles was calculated according to the following formula. The results are shown in Table 2. mRNA encapsulation rate (%) = (total mRNA concentration - mRNA concentration in the external aqueous phase) ÷ total mRNA concentration × 100

[0191] <Luciferase Luminescence Measurement> The mRNA lipid particle dispersion prepared in the above <Preparation of FLuc mRNA-Encapsulating Lipid Particles> was administered to ICR mice via the dorsal rectus femoris muscle in a single dose at an mRNA dose of 1 μg. Five hours and 50 minutes after administration, 150 mg / kg of D-luciferin potassium (Fujifilm Wako Pure Chemical Industries) was administered intraperitoneally. Six hours after administration, luminescence was measured in the prone position under isoflurane gas anesthesia using an IVIS Imaging System (PerkinElmer). An ROI was set to include the entire lower limb on the injected side, and the luminescence (Photones / Sec) was quantified using Living Image Software (PerkinElmer). Luciferase [P / S] in Table 2 indicates Photons / Sec (light intensity).

[0192] The results are shown in Table 2.

[0193]

[0194] The nucleic acid-lipid composition of the present invention was shown to have a high reporter protein expression rate.

[0195] (PTEN Antisense Oligonucleotide Information) PTEN (Phosphatase and Tensin Homolog Deleted from Chromosome 10) is an enzyme that catalyzes the dephosphorylation of phosphatidylinositol 3,4,5-triphosphate, an inositol phospholipid. Antisense oligonucleotide nucleic acid (PTEN ASO) against the PTEN protein was purchased from Hokkaido System Science Co., Ltd. It is a 20-base oligonucleotide with phosphodiester bonds, and its sequence is shown below. 5(m)^t(m)^g(m)^5(m)^t(m)^a^g^5c^5c^t^5c^t^g^g^a^t(m)^t(m)^t(m)^g(m)^a(m) where the lowercase letters (a, g, t) represent adenine, guanine, and thymidine, respectively, and (m) represents 2'MOE modification. 5(m) = 2'-MOE 5-Me cytosine, t(m) = 2'-MOE thymidine, g(m) = 2'-MOE guanine, a(m) = 2'-MOE adenine, 5c = 5-methyl-d cytosine, and ^ represents phosphorothioate. 2'-MOE represents 2'-O-methoxyethyl.

[0196] (Preparation of PTEN ASO-LNP) The first lipid, phospholipid, cholesterol, and polyethylene glycol lipid (PEG lipid) shown in Table 3 were dissolved in ethanol at the molar ratios shown in Table 3 to a total lipid concentration of 20 mmol / L to obtain an oil phase.

[0197] The neutral lipid used was DSPC (1,2-distearoyl-sn-glycero-3-phosphocholine, product name: COATSOME MC-8080; NOF Corporation). The cholesterol used was cholesterol (product name: Cholesterol HP; Nippon Fine Chemical Co., Ltd.). The PEG lipid used was monostearyl PEG (product name: Polyethylene Glycol Monostearate (4E.O.), Fujifilm Wako Pure Chemical Industries, Ltd.).

[0198] The structure of monostearyl PEG (also referred to as monoPEG) is shown below.

[0199] 5 mg of PTEN ASO was dissolved in 1 mL of sterile water and diluted with 10 mmol / L acetate buffer at pH 4 to a nucleic acid concentration of 54.6 μmol / L to obtain an aqueous phase. The aqueous and oil phases were then mixed using a syringe pump in a micromixer (see Japanese Patent No. 5288254) so ​​that the volume ratio of the aqueous phase to the oil phase was 3:1, and the mixture was diluted 2-fold with phosphate-buffered saline (PBS) to obtain a dispersion of nucleic acid-lipid particles.

[0200] Table 3 also shows the molar ratios of the first lipid, phospholipid, sterol, and PEG lipid in the lipid composition, and the mass ratio of nucleic acid to total lipid at the time of mixing.

[0201]

[0202] <Measurement of particle size> The particle size and polydispersity index of the lipid particles were measured using a zeta potential / particle size measurement system ELS-Z2 (Otsuka Electronics) after diluting the lipid particle dispersion 10 times with phosphate buffered saline (PBS). The measurement results are shown in Table 3.

[0203] <Evaluation of PTEN ASO Encapsulation Rate> (Quantification of Total Nucleic Acid Concentration) 30 μL of 3 mol / L aqueous sodium acetate solution and 9 μL of glycogen were added to 60 μL of lipid particles carrying nucleic acids, followed by the addition of 1.5 mL of ethanol to dissolve the lipids and precipitate only the nucleic acids. The mixture was then centrifuged and the supernatant was removed. After air-drying for 15 minutes or more, the mixture was redissolved in water and the total nucleic acid concentration was quantified by measuring the concentration using a Nanodrop ND1000 (Thermo Fisher Scientific).

[0204] (Quantification of nucleic acid concentration in the external aqueous phase) Quantitation was performed using the Quant-iT RiboGreen RNA Assay Kit (Thermo Fisher Scientific) according to the protocol. First, the 20x TE buffer included in the kit was diluted with water to obtain 1x TE buffer. TE stands for Tris / EDTA (ethylenediaminetetraacetic acid). To quantify only the nucleic acid in the external aqueous phase, the lipid particle dispersion containing nucleic acid was diluted 10,000 times with 1x TE buffer.

[0205] 100 μL of a lipid particle dispersion diluted 10,000-fold was placed in a 96-well plate, and then 100 μL of RiboGreen reagent (contained in the above-mentioned Quanti-iT Ribogreen RNA Assay Kit) diluted 2,000-fold with 1×TE buffer was added to the sample. Fluorescence (excitation wavelength: 485 nm, fluorescence wavelength: 535 nm) was measured using a plate reader Infinite F200 (TECAN) to quantify the nucleic acid concentration in the external aqueous phase.

[0206] (Calculation of Encapsulation Rate) Using the quantitative results of the total nucleic acid concentration and the nucleic acid concentration in the external aqueous phase obtained in the above steps, the nucleic acid encapsulation rate of the nucleic acid-lipid particles was calculated according to the following formula: Nucleic acid encapsulation rate (%) = (total nucleic acid concentration - nucleic acid concentration in the external aqueous phase) ÷ total nucleic acid concentration × 100 The calculation results are shown in Table 3.

[0207] <In vitro PTEN mRNA knockdown evaluation> (Cells used for evaluation) In the in vitro test using A431 cells (American Type Culture Collection), a culture medium was used that was a mixture of E-MEM (Gibco), FBS (fetal bovine serum) (Gibco), Penicillin-streptomycin (Gibco), and NEAA (Non-Essential Amino Acid) (Fujifilm Wako Pure Chemical Industries) in a ratio of 88:10:1:1.

[0208] In an in vitro test using SH-SY5Y cells (American Type Culture Collection), a culture medium was used, which was a mixture of E-MEM (Gibco), Ham's F12 (Gibco), FBS (Gibco), Penicillin-streptomycin (Gibco), and NEAA (Fujifilm Wako Pure Chemical Industries) in a ratio of 41.5:41.5:15:1:1, respectively.

[0209] (Quantification of PTEN mRNA by PCR) PTEN protein mRNA was measured using TaqMan® Fast Advanced Cells-to-CT. ＴＭ The protocol for the kit (Thermo Fisher Scientific) was followed. A dispersion of nucleic acid-lipid particles, naked ASO, or PBS, prepared to a final ASO concentration of 500 nmol / L, was added to A431 cells or SH-SY5Y cells. The cells were incubated at 37°C and 5% CO ２ After 24 hours of exposure under controlled conditions, the culture supernatant was removed and the cells were washed once with 4°C PBS. After removing the PBS, lysis solution was added at 50 μL / well and left to stand at room temperature for 5 minutes to obtain a cell lysate. The cell lysate was analyzed using TaqMan® Fast Advanced Cells-to-CT. ＴＭ Reverse transcription and PCR reactions were performed using the PCR Reagent Kit (Thermo Fisher Scientific), and PCR reagents Hs02621230_s1, FAM / MGB (Thermo Fisher Scientific), and Human GAPDH Endogenous Control, VIC® / MGB (Thermo Fisher Scientific).

[0210] The PTEN mRNA value of each sample was calculated using the ΔΔCt method. Specifically, the ΔCt value of each sample was calculated by subtracting the GAPDH Ct value from the PTEN Ct value. The average ΔCt value of the PBS-treated group was subtracted from the calculated ΔCt value to calculate the ddCt value. For each ΔΔCt value, a relative value to the naked ASO used as a comparison was taken, and this was used as the PTEN mRNA relative value. The calculation results are shown in Table 4.

[0211]

Claims

1. A compound represented by Formula (1) or a salt thereof,in the formula,R1 and R2 each independently represent a hydrocarbon group having 1 to 18 carbon atoms, and R3 represents a hydrocarbon group having 2 to 8 carbon atoms, where the hydrocarbon groups represented by R1, R2, and R3 may be substituted with one or more substituents selected from -OH, -COOH, -NR51R52, -OC(O)O-R53, -C(O)O-R54, -OC(O)-R55, and -O-R56,R4 represents a hydrocarbon group having 1 to 8 carbon atoms,R5 and R6 each independently represent a hydrocarbon group having 1 to 8 carbon atoms or -R8-L1-R9, excluding a case that both R5 and R6 are hydrocarbon groups having 1 to 8 carbon atoms,R7 represents -R10-L2-R11-L3-R12,R51 and R52 each independently represent a hydrocarbon group having 1 to 8 carbon atoms,R53, R54, R55, and R56 each independently represent a hydrocarbon group having 1 to 24 carbon atoms,the hydrocarbon groups represented by R53, R54, R55, and R56 may be substituted with an aryl group having 6 to 20 carbon atoms or -S-R58,the aryl group having 6 to 20 carbon atoms may be substituted with -OH, -COOH, -NR51R52, -OC(O)O-R53, -C(O)O-R54, -OC(O)-R55, -O-R56, or -(hydrocarbon group having 1 to 12 carbon atoms)-R57,R58 represents a hydrocarbon group having 1 to 12 carbon atoms,R57 represents -OH, -COOH, -NR61R62, -OC(O)O-R63, -C(O)O-R64, -OC(O)-R65, or -O-R66,R61 and R62 each independently represent a hydrocarbon group having 1 to 8 carbon atoms,2022263899   20 Mar 2026R63, R64, R65, and R66 each independently represent a hydrocarbon group having 1 to 24 carbon atoms,the hydrocarbon groups represented by R63, R64, R65, and R66 may be substituted with an aryl group having 6 to 20 carbon atoms or -S-R68,the aryl group having 6 to 20 carbon atoms may be substituted with -OH, -COOH, -NR61R62, -OC(O)O-R63, -C(O)O-R64, -OC(O)-R65, -O-R66, or -(hydrocarbon group having 1 to 12 carbon atoms)-R67,R68 represents a hydrocarbon group having 1 to 12 carbon atoms,L1, L2, and L3 each independently represent -OC(O)O-, -C(O)O-, -OC(O)-, or -O-,R8 represents a hydrocarbon group having 1 to 12 carbon atoms,R9 represents a hydrocarbon group having 1 to 24 carbon atoms,R10 represents a hydrocarbon group having 1 to 8 carbon atoms,R11 represents a hydrocarbon group having 1 to 24 carbon atoms,R12 represents a hydrocarbon group having 1 to 24 carbon atoms,the hydrocarbon groups represented by R9 and R12 may be substituted with an aryl group, -OC(O)O-R53, -C(O)O-R54, -OC(O)-R55, or -S-R58, where definitions of R53, R54, R55, and R58 are as described above, andthe hydrocarbon group represented by R11 may be substituted with -OC(O)O-R53, -C(O)O-R54, or -OC(O)-R55, where the definitions of R53, R54, and R55 are as described above.

2. The compound or a salt thereof according to claim 1, which is a compound represented by Formula (1-1) or a salt thereof,in the formula,R1 and R2 each independently represent a hydrocarbon group having 1 to 18 carbon atoms, and R3 represents a hydrocarbon group having 2 to 8 carbon atoms, where the hydrocarbon groups represented by R1, R2, and R3 may be substituted with -OH, -COOH, -NR51R52, -OC(O)O-R53, -C(O)O-R54, -OC(O)-R55, or -O-R56,2022263899   20 Mar 2026R4 represents a hydrocarbon group having 1 to 8 carbon atoms,R5 and R6 each independently represent a hydrocarbon group having 1 to 8 carbon atoms or -R8-L1-R9, excluding a case that both R5 and R6 are hydrocarbon groups having 1 to 8 carbon atoms,L1 represents -OC(O)O-, -C(O)O-, -OC(O)-, or -O-,R8 represents a hydrocarbon group having 1 to 12 carbon atoms,R9 represents a hydrocarbon group having 1 to 24 carbon atoms, where the hydrocarbon group represented by R9 may be substituted with an aryl group, -OC(O)O-R53, -C(O)O-R54, -OC(O)-R55, or -S-R58,R51 and R52 each independently represent a hydrocarbon group having 1 to 8 carbon atoms,R53, R54, R55, and R56 each independently represent a hydrocarbon group having 1 to 24 carbon atoms,the hydrocarbon groups represented by R53, R54, R55, and R56 may be substituted with an aryl group having 6 to 20 carbon atoms or -S-R58,the aryl group having 6 to 20 carbon atoms may be substituted with -OH, -COOH, -NR51R52, -OC(O)O-R53, -C(O)O-R54, -OC(O)-R55, -O-R56, or -(hydrocarbon group having 1 to 12 carbon atoms)-R57,R58 represents a hydrocarbon group having 1 to 12 carbon atoms,R57 represents -OH, -COOH, -NR51R52, -OC(O)O-R53, -C(O)O-R54, -OC(O)-R55, or -O-R56,R13 represents a hydrocarbon group having 1 to 8 carbon atoms,R14 represents -R15-L5-R16, where R15 represents a hydrocarbon group having 1 to 24 carbon atoms, L5 represents -OC(O)O-, -C(O)O-, -OC(O)-, or -O-, and R16 represents a hydrocarbon group having 1 to 24 carbon atoms,the hydrocarbon group having 1 to 24 carbon atoms represented by R15 may be substituted with -OC(O)O-R53, -C(O)O-R54, or -OC(O)-R55, where definitions of R53, R54, and R55 are as described above, andthe hydrocarbon group having 1 to 24 carbon atoms represented by R16 may be substituted with an aryl group having 6 to 20 carbon atoms, -OC(O)O-R53, -C(O)O-R54, -OC(O)-R55 or -S-R58, where the definitions of R53, R54, R55, and R58 are as described above.

3. The compound and a salt thereof according to claim 3,2022263899   20 Mar 2026wherein in Formula (1-1),R1 and R2 each independently represent a hydrocarbon group having 1 to 3 carbon atoms, and R3 represents a hydrocarbon group having 2 to 4 carbon atoms, where the hydrocarbon groups represented by R1, R2, and R3 may be substituted with -OH,R4 represents a hydrocarbon group having 1 to 8 carbon atoms,R5 and R6 each independently represent a hydrocarbon group having 1 to 8 carbon atoms or -R8-L1-R9, excluding a case that both R5 and R6 are hydrocarbon groups having 1 to 8 carbon atoms,L1 represents -C(O)O- or -OC(O)-,R8 represents a hydrocarbon group having 1 to 8 carbon atoms,R9 represents a hydrocarbon group having 1 to 18 carbon atoms, and the hydrocarbon group represented by R9 may be substituted with an aryl group having 6 to 20 carbon atoms or -S-R58,R58 represents a hydrocarbon group having 1 to 8 carbon atoms,R13 represents a hydrocarbon group having 1 to 8 carbon atoms,R14 represents -R15-L5-R16, where R15 represents a hydrocarbon group having 1 to 18 carbon atoms, L5 represents -OC(O)O-, and R16 represents a hydrocarbon group having 1 to 18 carbon atoms,the hydrocarbon group having 1 to 18 carbon atoms represented by R15 may be substituted with -C(O)O-R55 or -OC(O)-R56,R55 and R56 each independently represent a hydrocarbon group having 1 to 16 carbon atoms, and the hydrocarbon groups represented by R55 and R56 may be substituted with an aryl group having 6 to 20 carbon atoms or -S-R58, where the definition of R58 is as described above, andthe hydrocarbon group having 1 to 18 carbon atoms represented by R16 may be substituted with an aryl group or -S-R58, where the definition of R58 is as described above.

4. Lipid particles comprising:the compound or a salt thereof according to any one of claims 1 to 3; anda lipid.

5. The lipid particles according to claim 4, wherein the lipid is at least one kind of lipid selected from the group consisting of a2022263899   20 Mar 2026sterol and a lipid having a nonionic hydrophilic polymer chain.

6. The lipid particles according to claim 4 or 5, further comprising: a neutral lipid.

7. The lipid particles according to any one of claims 4 to 6, further comprising: a nucleic acid.

8. The lipid particles according to claim 7,wherein the nucleic acid includes a nucleic acid having 50 or more bases.

9. A pharmaceutical composition comprising:the lipid particles according to any one of claims 4 to 8 as an active ingredient.