Tricin and citrate-based cationic lipids

Abstract

The present invention provides, in part, tricine and citrate lipids, or pharmaceutically acceptable salts thereof. The compounds provided herein, comprising compounds of formula (A), may be useful for the delivery and expression of mRNA and encoded proteins, for example, as components of liposome delivery vehicles, and therefore may be useful in treating a variety of diseases, disorders and conditions, including those associated with the deficiency of one or more proteins. [Case 1] TIFF2026524782000711.tif22170

Description

[Technical Field]

[0001] Related applications This application claims priority to European Patent Application No. 23305930.2 filed on 12 June 2023, European Patent Application No. 23306900.4 filed on 2 November 2023, and European Patent Application No. 23306902.0 filed on 2 November 2023, the entirety of which disclosures are incorporated herein by reference. [Background technology]

[0002] Nucleic acid delivery is being widely studied as a potential treatment option for certain disease conditions. In particular, messenger RNA (mRNA) therapy is becoming an increasingly important option for treating a variety of diseases, including those associated with the deficiency of one or more proteins. [Overview of the Initiative] [Means for solving the problem]

[0003] The present invention provides, in particular, a novel class of tricine and citrate-based cationic lipid compounds for improved in vivo delivery of therapeutic agents, such as nucleic acids. The compounds provided herein are thought to enable highly effective in vivo delivery while maintaining a favorable toxicity profile. The compounds provided herein are thought to enable highly effective intranasal delivery. In fact, Examples 43 and 45 demonstrate that lipid nanoparticles containing the compounds of the present invention (e.g., compound 64 or the compounds in Table 6, or pharmaceutically acceptable salts thereof) are particularly effective for intranasal delivery.

[0004] In one embodiment, the present invention relates to a structure of formula (A2): [ka] The formula is characterized by a cationic lipid having or a pharmaceutically acceptable salt thereof, wherein X is independently either O or NH; R , , , , , 3 , 3 , 1 , 4 , 10 , 5 , 24 , , , , , 2 , 5 , 30 , 2 , 10 , 3 , 10 , R 2 , and R 3 Each of which is independently C4 - C 30 alkyl, C4 - C 30 alkenyl, C4 - C 30 alkynyl, or C4 - C 30 heteroalkyl; A is

Chemical formula

[0005] In one embodiment, the present invention relates to a structure of formula (A1): [ka] The formula is characterized by a cationic lipid having or a pharmaceutically acceptable salt thereof, wherein X is independently either O or NH; R 1 , R 2 , and R 3 Each of these is independent of C4~C 30 Alkyl, C4~C 30 Alkenyl, C4~C 30 Alkinyl, or C4~C 30 It is heteroalkyl; A is [ka] It is a substructure selected from, B is independently an ionizable nitrogen-containing group or a permanently charged nitrogen group; m is an integer between 2 and 10; n is an integer between 2 and 10; L 1 is a carbonyl, ester, or amide; L 2 C2~C 10 Alkylene or C2~C 10 It is an alkenylene; Ar is a phenylene compound optionally containing 1 to 4 substituents independently selected from halogens, OCH3, and CH3; R 4 C1~C 10 It is alkylene; Here, A is R 1 , R 2 , and R 3 Each of them independently contains a disulfide bond, or [ka] C4~C has the structure 30If it is a heteroalkyl, it is the only substructure (a4), L 3 is OC(O), CO2, or (O)CO; o is an integer between 2 and 5; R 5 C4~C 24 Alkyl; or L 3 is OC(O), CO2, or (O)CO; o is an integer between 6 and 12; R 5 These are C1-C5 alkyl groups.

[0006] In one embodiment, the present invention relates to a structure of formula (A): [ka] The formula is characterized by a cationic lipid having or a pharmaceutically acceptable salt thereof, wherein X is independently either O or NH; R 1 , R 2 , and R 3 Each of these independently corresponds to C4~C 30 Alkyl, C4~C 30 Alkenyl, C4~C 30 Alkinyl, or C4~C 30 It is heteroalkyl; A is [ka] It is a substructure selected from, B is independently an ionizable nitrogen-containing group or a permanently charged nitrogen group; m is an integer between 2 and 10; n is an integer between 2 and 10; L 1 is a carbonyl, ester, or amide; L 2 C2~C 10 Alkylene or C2~C 10 It is an alkenylene; Ar is a phenylene compound optionally containing 1 to 4 substituents independently selected from halogens, OCH3, and CH3; R 4 C2~C10 It is alkylene; Here, A is R 1 , R 2 , and R 3 Each of them independently contains a disulfide bond, or [ka] C4~C has the structure 30 If it is a heteroalkyl, it is the only substructure (a4), L 3 is OC(O), CO2, or (O)CO; o is an integer between 2 and 5; R 5 C4~C 24 Alkyl; or L 3 is OC(O), CO2, or (O)CO; o is an integer between 6 and 12; R 5 These are C1-C5 alkyl groups.

[0007] In this embodiment, X is O, and / or B is independently an ionizable nitrogen-containing group.

[0008] In this embodiment, R 1 , R 2 , and R 3 Each of these is independent of C4~C 30 Alkyl, C4~C 30 Alkenyl, or C4~C 30 It is alkinyl.

[0009] In the embodiment of formula (A2), R 1 , R 2 , and R 3 Each of them independently contains a disulfide bond, or [ka] C4~C has the structure 30 It is a heteroalkyl, where L 3 is OC(O), CO2, or (O)CO; o is an integer between 2 and 5; R 5 is C4~C 24 Alkyl; or L 3is OC(O), CO2, or (O)CO; o is an integer from 6 to 12; R 5 is C1-C6 alkyl.

[0010] In an embodiment, R 1 , R 2 , and R 3 each independently contains a disulfide bond or

Chemical formula

Chemical formula

[0016] In an embodiment, B is independently an ionizable nitrogen-containing group.

[0017] In an embodiment, m is 2 and / or n is 2 or 3.

[0018] In an embodiment, B is independently

Chemical formula

[0019] In an embodiment, B is independently

Chemical formula

[0020] In an embodiment, B is independently

Chemical formula

[0021] In an embodiment, B is independently

Chemical formula

[0022] In this embodiment, B is [ka] That is the case.

[0023] In this embodiment, the compound is selected from the group consisting of compounds (1), (2), (3), (4), (6), (9), (17), (24), (26), (27), (33), and (35).

[0024] In the embodiment, the compound has the structure of formula (III): [ka] A salt having or a pharmaceutically acceptable salt thereof, in the formula, R 1 , R 2 , and R 3 Each of these is independent of C6~C 30 Alkyl, C6~C 30 Alkenil, C6~C 30 Alkinyl, or C4~C 30 It is heteroalkyl; R 4 C2~C 10 It is alkylene; B is an independently ionizable nitrogen-containing group.

[0025] In this embodiment, R 4 It is -CH2CH2-.

[0026] In this embodiment, B is independently, [ka] That is the case.

[0027] In this embodiment, B is independently, [ka] That is the case.

[0028] In this embodiment, B is independently, [ka] That is the case.

[0029] In this embodiment, B is independently, [ka] That is the case.

[0030] In this embodiment, B is [ka] That is the case.

[0031] In this embodiment, the compound is compound (3).

[0032] In the embodiment, the compound has the structure of formula (IV): [ka] A salt having or a pharmaceutically acceptable salt thereof, in the formula, R 1 , R 2 , and R 3 Each of these is independent of C6~C 30 Alkyl, C6~C 30 Alkenil, C6~C 30 Alkinyl, or C4~C 30 It is heteroalkyl; m is an integer between 2 and 10; n is an integer between 2 and 10; B is an independently ionizable nitrogen-containing group.

[0033] In this embodiment, R 1 , R 2 , and R 3 C4~C 30 It is heteroalkyl.

[0034] In the embodiment, m is 2 and / or n is 2 or 3.

[0035] In this embodiment, B is independently, [ka] That is the case.

[0036] In this embodiment, B is independently, [ka] That is the case.

[0037] In this embodiment, B is independently, [ka] That is the case.

[0038] In this embodiment, B is independently, [ka] That is the case.

[0039] In this embodiment, B is [ka] That is the case.

[0040] In the embodiment, the compound is compound (8) or (39).

[0041] In the embodiment, the compound has the structure of formula (V1): [ka] A salt having or a pharmaceutically acceptable salt thereof, in the formula, n is an integer of 2, 3, 4, 5, 6, or 7; B is an independently ionizable nitrogen-containing group.

[0042] In this embodiment, L 3 is OC(O), CO2, or (O)CO; o is an integer between 2 and 5; R 5 C4~C 24 It is alkyl.

[0043] In the embodiments of formula (A1), formula (A), and formula (V1), L 3 is OC(O), CO2, or (O)CO; o is an integer between 6 and 12; R 5 is a C1-C5 alkyl group. In embodiments of formula (A2) and formula (V1), L 3 is OC(O), CO2, or (O)CO; o is an integer between 6 and 12; R 5 These are C1-C6 alkyl groups.

[0044] In this embodiment, B is independently, [ka] That is the case.

[0045] In this embodiment, B is independently, [ka] That is the case.

[0046] In this embodiment, B is independently, [ka] That is the case.

[0047] In this embodiment, B is independently, [ka] That is the case.

[0048] In this embodiment, B is [ka] That is the case.

[0049] In the embodiment, the compound is selected from the group consisting of compounds (11), (12), (13), (14), (22), (25), (28), (29), (30), (32), (34), (36), (38), (40), (112), (113), (114), (115), (116), (117), (126), (127), (128), and (129).

[0050] In the embodiment, the compound has the structure of formula (V): [ka] A salt having or a pharmaceutically acceptable salt thereof, in the formula, n is an integer of 2, 3, or 4; B is an independently ionizable nitrogen-containing group.

[0051] In this embodiment, L 3 is OC(O), CO2, or (O)CO; o is an integer between 2 and 5; R 5 C4~C 24 It is alkyl.

[0052] In the embodiments of formula (A1), formula (A), and formula (V), L 3 is OC(O), CO2, or (O)CO; o is an integer between 6 and 12; R 5 is a C1-C5 alkyl group. In embodiments of formula (A2) and formula (V), L 3 is OC(O), CO2, or (O)CO; o is an integer between 6 and 12; R 5 These are C1-C6 alkyl groups.

[0053] In this embodiment, B is independently, [ka] That is the case.

[0054] In this embodiment, B is independently, [ka] That is the case.

[0055] In this embodiment, B is independently, [ka] That is the case.

[0056] In this embodiment, B is independently, [ka] That is the case.

[0057] In this embodiment, B is [ka] That is the case.

[0058] In the embodiment, the compound is selected from the group consisting of compounds (11), (12), (13), (14), (22), (25), (28), (29), (30), (32), (34), (36), (38), (40), (114), and (116).

[0059] In this embodiment, the compound is selected from the group consisting of compounds (11), (12), (13), (14), (22), (25), (28), (29), (30), (32), (34), (36), (38), and (40).

[0060] In the embodiment, the compound has the structure of formula (VI): [ka] A salt having or a pharmaceutically acceptable salt thereof, in the formula, R 1 , R 2 , and R 3 Each of them independently contains a disulfide group C6-C 30 It is heteroalkyl; n is an integer of 2, 3, or 4; B is an independently ionizable nitrogen-containing group.

[0061] In this embodiment, B is independently, [ka] That is the case.

[0062] In this embodiment, B is independently, [ka] That is the case.

[0063] In this embodiment, B is independently, [ka] That is the case.

[0064] In this embodiment, B is independently, [ka] That is the case.

[0065] In this embodiment, B is [ka] That is the case.

[0066] In this embodiment, the compound is compound (21).

[0067] In this embodiment, the compound is related to formula (VII): [ka] A salt having or a pharmaceutically acceptable salt thereof, in the formula, X is independently either O or NH; R 1 , R 2 , and R 3Each of these independently corresponds to C4~C 30 Alkyl, C4~C 30 Alkenyl, C4~C 30 Alkinyl, or C4~C 30 It is heteroalkyl; Each m is an integer between 2 and 10; Each L 4 is a carbonyl, ester, or amide; Z is -(CH2) q1 -N-(CH2) q2 -where q1 and q2 are independently integers between 2 and 10; or Z is C6H3-Z 1 And here, Z 1 is -CH2(CH2) q The B portion is covalently bonded to a carbonyl, ester, or amide; q is an integer between 1 and 9; B is an independently ionizable nitrogen-containing group.

[0068] In the embodiment, X is O; each L 4 is -C(O)O-; m is an integer of 2, 3, or 4; and / or q is 1.

[0069] In this embodiment, Z is -(CH2) q1 -N-(CH2) q2 - and q1 and q2 are both 2.

[0070] In this embodiment, Z is [ka] That is the case.

[0071] In this embodiment, B is independently, [ka] That is the case.

[0072] In this embodiment, B is independently, [ka] That is the case.

[0073] In this embodiment, B is independently, [ka] That is the case.

[0074] In this embodiment, B is independently, [ka] That is the case.

[0075] In this embodiment, B is [ka] That is the case.

[0076] In the embodiment, the compound is compound (16) or (31).

[0077] In one embodiment, the present invention is characterized by a method for delivering a composition comprising mRNA encoding a protein or polypeptide encapsulated within a liposome to the lung, intranasal cavity, or intramuscular muscle, wherein the liposome comprises one or more cationic lipids, optionally one or more non-cationic lipids, optionally one or more cholesterol-based lipids, and optionally one or more PEG-modified lipids, wherein at least one cationic lipid is a compound having a structure relating to formula (A2), formula (A1), or formula (A) as described herein, or a pharmaceutically acceptable salt thereof.

[0078] In one embodiment, the present invention is characterized by a method of delivering a composition containing nucleic acids encapsulated in liposomes to the lungs, nasal cavity, or intramuscular muscle, wherein the liposomes contain cationic lipids which are compounds having a structure relating to formula (A2), formula (A1), or formula (A) as described herein, or pharmaceutically acceptable salts thereof.

[0079] In one embodiment, the present invention is characterized by a method of intranasal delivery of a composition comprising mRNA encoding a protein or polypeptide encapsulated within a liposome, wherein the liposome comprises one or more cationic lipids, optionally one or more non-cationic lipids, optionally one or more cholesterol-based lipids, and optionally one or more PEG-modified lipids, wherein at least one cationic lipid is a compound having a structure relating to formula (A2), formula (A1), or formula (A) as described herein, or a pharmaceutically acceptable salt thereof, for example, compound 64, or a compound in Table 6, or a pharmaceutically acceptable salt thereof.

[0080] In one embodiment, the present invention is characterized by a method for intranasal delivery of a composition containing nucleic acids encapsulated in liposomes, wherein the liposomes contain cationic lipids which are compounds having a structure relating to formula (A2), formula (A1), or formula (A) as described herein, or pharmaceutically acceptable salts thereof, such as compound 64, or compounds in Table 6, or pharmaceutically acceptable salts thereof.

[0081] In another embodiment, the compound is selected from the group of compounds 41-64 and 90, or pharmaceutically acceptable salts thereof.

[0082] [Table 1]

[0083] [Table 2]

[0084] [Table 3]

[0085] [Table 4]

[0086] [Table 5]

[0087] [Table 6]

[0088] [Table 7]

[0089] In another embodiment, the compound is selected from the group of compounds 91 to 111 or pharmaceutically acceptable salts thereof.

[0090] [Table 8]

[0091] [Table 9]

[0092] [Table 10]

[0093] [Table 11]

[0094] [Table 12]

[0095] In another embodiment, the compound is selected from the group of compounds 112-117 and 126-129, or pharmaceutically acceptable salts thereof.

[0096] [Table 13]

[0097] [Table 14]

[0098] [Table 15]

[0099] In another embodiment, the compound is selected from the group of compounds 118-125 or pharmaceutically acceptable salts thereof.

[0100] [Table 16]

[0101] [Table 17]

[0102] [Table 18]

[0103] [Table 19]

[0104] In embodiments, the composition comprises mRNA encoding a protein or polypeptide encapsulated within a liposome, the liposome comprising one or more cationic lipids, optionally one or more non-cationic lipids, optionally one or more cholesterol-based lipids, and optionally one or more PEG-modified lipids, the at least one cationic lipid being any cationic lipid described herein.

[0105] In the embodiments, the composition comprises nucleic acids encapsulated within liposomes, the liposomes comprising cationic lipids, which are any cationic lipids described herein.

[0106] In the embodiment, the composition further comprises one or more cationic lipids, one or more non-cationic lipids, and one or more PEG-modified lipids, plus another lipid selected from the group.

[0107] In the embodiment, the nucleic acid is mRNA that encodes a peptide or polypeptide.

[0108] In this embodiment, mRNA encoding a peptide or polypeptide for use in a vaccine.

[0109] In this embodiment, the mRNA encodes an antigen.

[0110] In this embodiment, the antigen is derived from an infectious agent.

[0111] In embodiments, the composition is formulated for administration routes including oral, rectal, vaginal, mucosal, intrapulmonary, or intestinal administration; intradermal, transdermal (topical), intramuscular, subcutaneous, intrathecal injection; and parenteral delivery including intrathecal, direct intraventricular, intravenous, intraperitoneal, or nasal delivery.

[0112] In this embodiment, lung delivery is intratracheal or by inhalation.

[0113] In one embodiment, the route of administration is intranasal.

[0114] In this embodiment, the route of administration is intramuscular.

[0115] In this embodiment, the route of administration is intrapulmonary.

[0116] In another embodiment, the present invention is characterized by a method for delivering a composition comprising mRNA encoding a protein or polypeptide encapsulated within a liposome to the lung, intranasal cavity, or intramuscular muscle, wherein the liposome comprises one or more cationic lipids, optionally one or more non-cationic lipids, optionally one or more cholesterol-based lipids, and optionally one or more PEG-modified lipids, wherein at least one cationic lipid has a structure relating to formula (B): [ka] A compound having, or a pharmaceutically acceptable salt thereof, wherein, Each n is independently either 0 or 1; X 1A Independently, O or NR 1A and; R 1A is H or C1-C6 alkyl, X 1B is a covalent bond, C(O), CH2CO2, or CH2C(O); X 2A and X 2B One of them is O, and the other is a covalent bond; X 3A and X 3B One of them is O, and the other is a covalent bond; X 4A and X 4B One of them is O, and the other is a covalent bond; R 1 Independently, L 1 -B 1 , C6~C 30 Alkyl, C6~C 30 Alkenyl, or C6~C 30 It is alkinyl; R 2 Independently, L 2 -B 2 , C6~C 30 Alkyl, C6~C 30 Alkenyl, or C6~C 30 It is alkinyl; R 3 Independently, L 3 -B 3 , C6~C 30 Alkyl, C6~C 30 Alkenyl, or C6~C 30 It is alkinyl; R 4 Independently, L 4 -B 4 , C6~C 30 Alkyl, C6~C 30 Alkenyl, or C6~C 30 It is alkinyl; L1 , L 2 , L 3 , and L 4 Each of these is independent of C1~C 30 Alkylene; C2~C 30 Alkenylene; or C2~C 30 It is alkynylene; B 1 B 2 B 3 , and B 4 Each of them is independently an ionizable nitrogen-containing group, Cationic lipids contain at least one ionizable nitrogen-containing group.

[0117] In another aspect, the present invention is characterized by a method for delivering a composition comprising nucleic acid encapsulated in liposomes to the lungs, nasal cavity, or muscle, wherein the liposome has a structure relating to formula (B): [ka] A compound having, or a cationic lipid which is a pharmaceutically acceptable salt thereof, in the formula, Each n is independently either 0 or 1; X 1A Independently, O or NR 1A and; R 1A is H or C1-C6 alkyl, X 1B is a covalent bond, C(O), CH2CO2, or CH2C(O); X 2A and X 2B One of them is O, and the other is a covalent bond; X 3A and X 3B One of them is O, and the other is a covalent bond; X 4A and X 4B One of them is O, and the other is a covalent bond; R 1 Independently, L 1 -B 1 , C6~C 30 Alkyl, C6~C 30Alkenyl, or C6~C 30 It is alkinyl; R 2 Independently, L 2 -B 2 , C6~C 30 Alkyl, C6~C 30 Alkenyl, or C6~C 30 It is alkinyl; R 3 Independently, L 3 -B 3 , C6~C 30 Alkyl, C6~C 30 Alkenyl, or C6~C 30 It is alkinyl; R 4 Independently, L 4 -B 4 , C6~C 30 Alkyl, C6~C 30 Alkenyl, or C6~C 30 It is alkinyl; L 1 , L 2 , L 3 , and L 4 Each of these is independent of C1~C 30 Alkylene; C2~C 30 Alkenylene; or C2~C 30 It is alkynylene; B 1 B 2 B 3 , and B 4 Each of them is independently an ionizable nitrogen-containing group, Cationic lipids contain at least one ionizable nitrogen-containing group.

[0118] In another embodiment, the present invention is characterized by a method for delivering a composition comprising mRNA encoding a protein or polypeptide encapsulated within a liposome to the lung, intranasal cavity, or intramuscular muscle, wherein the liposome comprises one or more cationic lipids, optionally one or more non-cationic lipids, optionally one or more cholesterol-based lipids, and optionally one or more PEG-modified lipids, the at least one cationic lipid having a structure relating to formula (B1): [ka] A compound having the following characteristics, in which B 1 teeth, [ka] and; L 1 It is a straight-chain C5 alkylene; R 2 , R 3 , and R 4 This is as defined for equation (B).

[0119] In another embodiment, the present invention is characterized by a method for delivering a composition comprising mRNA encoding a protein or polypeptide encapsulated within a liposome to the lung, intranasal cavity, or intramuscular muscle, wherein the liposome comprises one or more cationic lipids, optionally one or more non-cationic lipids, optionally one or more cholesterol-based lipids, and optionally one or more PEG-modified lipids, the at least one cationic lipid having a structure relating to formula (B1): [ka] A compound having the following characteristics, in which B 1 teeth, [ka] and; L 1 This is defined for equation (B); R 2 , R 3 , and R 4 It is a straight-chain C8 alkylene.

[0120] In another aspect, the present invention is characterized by a method for delivering a composition comprising nucleic acid encapsulated in liposomes to the lungs, nasal cavity, or muscle, wherein the liposome has a structure relating to formula (B1): [ka] The compound contains a cationic lipid which has B in its formula, 1teeth, [ka] and; L 1 This is defined for equation (B); R 2 , R 3 , and R 4 It is a straight-chain C8 alkylene.

[0121] In another aspect, the present invention is characterized by a method for delivering a composition comprising nucleic acid encapsulated in liposomes to the lungs, nasal cavity, or muscle, wherein the liposome has a structure relating to formula (B1): [ka] The compound contains a cationic lipid which has B in its formula, 1 teeth, [ka] and; L 1 This is defined for equation (B); R 2 , R 3 , and R 4 It is a linear C8 alkylene.

[0122] In embodiments, the present invention also provides compounds of formula (B1) as described herein. These compounds may be incorporated into any of the compositions disclosed herein and may be used in any of the methods described herein.

[0123] In the embodiment, the composition further comprises one or more cationic lipids, one or more non-cationic lipids, and one or more PEG-modified lipids, plus another lipid selected from the group.

[0124] In the embodiment, the nucleic acid is mRNA that encodes a peptide or polypeptide.

[0125] In the embodiment, the composition comprises mRNA encoding a peptide or polypeptide for use in a vaccine.

[0126] In this embodiment, the mRNA encodes an antigen.

[0127] In this embodiment, the antigen is derived from an infectious agent.

[0128] In the embodiment, the compound is selected from the group consisting of compounds (65) to (89) or pharmaceutically acceptable salts thereof.

[0129] [Table 20]

[0130] [Table 21]

[0131] [Table 22]

[0132] [Table 23]

[0133] [Table 24]

[0134] [Table 25]

[0135] [Table 26]

[0136] In the embodiment, the compound is selected from the group consisting of compounds (118) to (125) or pharmaceutically acceptable salts thereof.

[0137] [Table 27]

[0138] [Table 28]

[0139] [Table 29]

[0140] In this embodiment, delivery is intranasal.

[0141] In this embodiment, delivery is intramuscular.

[0142] In this embodiment, delivery takes place within the lungs. [Brief explanation of the drawing]

[0143] [Figure 1] The survival percentage of BALB / c mice inoculated with a phosphate-buffered saline (PBS) buffer control administered intranasally (IN) with lipid nanoparticles (LNPs) encapsulating CA09 HA mRNA (these LNPs contained either compound 64 or lipid OF-02); 2) intramuscularly (IM) with LNPs encapsulating CA09 HA mRNA (these LNPs contained only OF-02); or 3) intranasally (IN) is shown. These results demonstrate the high efficacy of LNPs containing compound 64 administered intranasally in a lethal mouse influenza inoculation model. [Figure 2]The results show the percentage change in body weight of BALB / c mice inoculated with a phosphate-buffered saline (PBS) buffer control administered intranasally (IN) with lipid nanoparticles (LNPs) encapsulating CA09 HA mRNA (these LNPs contained either compound 64 or lipid OF-02); intramuscularly (IM) with LNPs encapsulating CA09 HA mRNA (these LNPs contained only OF-02); or intranasally (IN). These results demonstrate the high efficacy of LNPs containing compound 64 administered intranasally in a lethal mouse influenza inoculation model. The data shown in the graph in Figure 2 spans 14 days after inoculation. [Figure 3] The following data are obtained from various studies comparing intranasal administration of compound 64 (the compound of the present invention) with intramuscular administration of the known compound OF-02, which is described in detail in Example 47 of this specification. [Figure 4] The following data are obtained from various studies comparing intranasal administration of compound 64 (the compound of the present invention) with intramuscular administration of the known compound OF-02, which is described in detail in Example 47 of this specification. [Modes for carrying out the invention]

[0144] definition To facilitate understanding of the present invention, certain terms are first defined below. Further definitions of the following terms and other terms are given throughout this specification. Publications and other reference materials referred to herein to describe the background of the present invention and to provide additional details relating to its implementation are incorporated herein by reference.

[0145] Amino Acids: As used herein, the term “amino acid” in its broadest sense refers to any compound and / or substance that can be incorporated into a polypeptide chain. In some embodiments, an amino acid has the general structure H2N-C(H)(R)-COOH. In some embodiments, an amino acid is a naturally occurring amino acid. In some embodiments, an amino acid is a synthetic amino acid; in some embodiments, an amino acid is a d-amino acid; in some embodiments, an amino acid is an l-amino acid. “Standard amino acid” refers to any of the 20 standard l-amino acids commonly found in naturally occurring peptides. “Non-standard amino acid” refers to any amino acid that is not a standard amino acid, whether it is synthetically prepared or obtained from a natural source. As used herein, “synthetic amino acid” includes, but is not limited to, chemically modified amino acids, including salts, amino acid derivatives (such as amides), and / or substitutions. Amino acids, including carboxy-terminal and / or amino-terminal amino acids in peptides, can be modified by methylation, amidation, acetylation, protecting groups, and / or substitution with other chemical groups that can alter the cyclic half-life of the peptide without adversely affecting its activity. Amino acids can be involved in disulfide bonds. Amino acids may have one or more posttranslational modifications, such as association with one or more chemical entities (e.g., methyl group, acetate group, acetyl group, phosphate group, formyl moiety, isoprenoid group, sulfate group, polyethylene glycol moiety, lipid moiety, carbohydrate moiety, biotin moiety, etc.). The term "amino acid" is used interchangeably with "amino acid residue" and may refer to free amino acids and / or amino acid residues of peptides. Whether it refers to free amino acids or peptide residues should be clear from the context in which the term is used.

[0146] Animals: As used herein, the term “animal” refers to any member of the animal kingdom. In some embodiments, “animal” refers to a human at any developmental stage. In some embodiments, “animal” refers to a non-human animal at any developmental stage. In certain embodiments, non-human animals are mammals (e.g., rodents, mice, rats, rabbits, monkeys, dogs, cats, sheep, cattle, primates, and / or pigs). In some embodiments, animals include, but are not limited to, mammals, birds, reptiles, amphibians, fish, insects, and / or worms. In some embodiments, animals may be transgenic animals, genetically modified animals, and / or clones.

[0147] Approximately or about: As used herein, the terms “approximately” or “about” applied to one or more values ​​of interest refer to values ​​that are similar to the stated reference values. In certain embodiments, unless otherwise specified or evident from the context, the terms “approximately” or “about” refer to values ​​that fall within a range of 25%, 20%, 19%, 18%, 17%, 16%, 15%, 14%, 13%, 12%, 11%, 10%, 9%, 8%, 7%, 6%, 5%, 4%, 3%, 2%, 1%, or less in any direction of the stated reference value (except where such numbers exceed 100% of the possible values).

[0148] Biologically active: As used herein, the term “biologically active” refers to the characteristic of any drug that is active in a biological system and, in particular, in a living organism. For example, a drug that has a biological effect on an organism when administered to that organism is considered biologically active.

[0149] Delivery: As used herein, the term “delivery” encompasses both local delivery and systemic delivery. For example, mRNA delivery includes situations in which mRNA is delivered to a target tissue, the encoded protein is expressed, and it is retained within the target tissue (also referred to as “local distribution” or “local delivery”) and situations in which mRNA is delivered to a target tissue, the encoded protein is expressed, it is secreted into the patient’s circulatory system (e.g., serum), systematically distributed, and absorbed by other tissues (also referred to as “systemic distribution” or “systemic delivery”).

[0150] Expression: As used herein, “expression” of a nucleic acid sequence refers to the translation of mRNA into polypeptides, the assembly of multiple polypeptides into an intact protein (e.g., an enzyme), and / or post-translational modification of polypeptides or fully assembled proteins (e.g., enzymes). In this application, the terms “expression” and “production” and their grammatical synonyms are used interchangeably.

[0151] Functional: As used herein, a “functional” biomolecule is a biomolecule in which it exhibits the properties and / or activity that characterize it.

[0152] Half-life: As used herein, the term “half-life” refers to the time required for an amount of a nucleic acid or protein, such as its concentration or activity, to fall to half of its initial measured value over a given period.

[0153] Helper Lipids: As used herein, the term “Helper Lipids” refers to any neutral or zwitterionic lipid material, including cholesterol. Without being constrained by any particular theory, helper lipids can impart stability, rigidity, and / or fluidity within lipid bilayers / nanoparticles.

[0154] Improvement, Increase, or Decrease: As used herein, the terms “improve,” “increase,” or “decrease,” or their grammatical synonyms, refer to a relative value to a baseline measurement, such as a measurement in the same individual prior to the initiation of the treatment described herein, or a measurement in a control subject (or control subject) that has not received the treatment described herein. A “control subject” is a subject suffering from the same form of disease as the subject under treatment and being of approximately the same age as the subject under treatment.

[0155] In vitro: As used herein, the term “in vitro” refers to an event that occurs in an artificial environment, such as in a test tube or reaction vessel, or in a cell culture, rather than within a multicellular organism.

[0156] In vivo: As used herein, the term “in vivo” refers to events occurring within multicellular organisms such as humans and non-human animals. In the context of cell-based systems, the term may be used to refer to events occurring within living cells (as opposed to, for example, in vitro systems).

[0157] Isolation: As used herein, the term “isolated” means a substance and / or entity that has been (1) separated from at least some of the components with which it was originally formed (whether naturally occurring or produced in an experimental setting), and / or (2) produced, prepared and / or manufactured by human hands. An isolated substance and / or entity may be separated from about 10%, about 20%, about 30%, about 40%, about 50%, about 60%, about 70%, about 80%, about 90%, about 91%, about 92%, about 93%, about 94%, about 95%, about 96%, about 97%, about 98%, about 99%, or more than about 99% of the other components with which it was originally formed. In some embodiments, the isolated agent is of a purity of about 80%, about 85%, about 90%, about 91%, about 92%, about 93%, about 94%, about 95%, about 96%, about 97%, about 98%, about 99%, or greater than about 99%. As used herein, a substance is "pure" if it is substantially free of other components. As used herein, the calculation of the purity percentage of an isolated substance and / or entity should not include excipients (e.g., buffers, solvents, water, etc.).

[0158] Liposome: As used herein, the term “liposome” refers to any layer, multilayer, or solid nanoparticle vesicle. Typically, liposomes as used herein may be formed by mixing one or more lipids or by mixing one or more lipids with a polymer. In some embodiments, liposomes suitable for the present invention contain one or more cationic lipids and optionally one or more non-cationic lipids, optionally one or more cholesterol-based lipids and / or optionally one or more PEG-modified lipids.

[0159] Messenger RNA (mRNA): As used herein, the term “messenger RNA (mRNA)” or “mRNA” refers to a polynucleotide encoding at least one polypeptide. As used herein, mRNA encompasses both modified and unmodified RNA. The term “modified mRNA” refers to mRNA containing at least one chemically modified nucleotide. mRNA may contain one or more coding and non-coding regions. mRNA may be purified from natural sources, produced using recombinant expression systems, and optionally purified, chemically synthesized, etc. Where appropriate, for example, in chemically synthesized molecules, mRNA may contain nucleoside analogs such as chemically modified bases or sugars, or analogs with skeletal modifications, etc. mRNA sequences are presented in the 5' to 3' direction unless otherwise indicated. In some embodiments, mRNA is a natural nucleoside (e.g., adenosine, guanosine, cytidine, uridine); nucleoside analogs (e.g., 2-aminoadenosine, 2-thiothymidine, inosine, pyrrolo-pyrimidine, 3-methyladenosine, 5-methylcytidine, C-5-propynylcytidine, C-5-propynyluridine, 2-aminoadenosine, C5-bromouridine, C5-fluorouridine, C5-iodouridine, C5-propynyluridine, C5-propynylcytidine, C5-methylcyt5-propynyluridine, C5-methylcytidine, C5-methylcytidine, C5-aminoadenosine, C5-5-propynyluridine, C5-methylcytidine, C5-aminoadenosine, C5-5-propynyluridine, C5-methylcytidine, C5-aminoadenosine, C5-5-propynyluridine, C5-methylcytidine, C5-aminoadenosine, C5-5-propynyluridine, C5-methylcytidine, C5-aminoadenosine Noadenosine, 7-deazaadenosine, 7-deazaguanosine, 8-oxoadenosine, 8-oxoguanosine, O(6)-methylguanine, and 2-thiocytidine); chemically modified bases; biologically modified bases (e.g., methylated bases); intercalated bases; modified sugars (e.g., 2'-fluororibose, ribose, 2'-deoxyribose, arabinose, and hexose); and / or modified phosphate groups (e.g., phosphorothioates and 5'-N-phosphoramidite bonds) or comprising them.

[0160] Nucleic Acids: As used herein, the term “nucleic acid” in its broadest sense refers to any compound and / or substance that is incorporated into or can be incorporated into a polynucleotide chain. In some embodiments, a nucleic acid is a compound and / or substance that is incorporated into or can be incorporated into a polynucleotide chain by phosphate diester bonds. In some embodiments, “nucleic acid” refers to individual nucleic acid residues (e.g., nucleotides and / or nucleosides). In some embodiments, “nucleic acid” refers to a polynucleotide chain comprising individual nucleic acid residues. In some embodiments, “nucleic acid” includes RNA and single-stranded and / or double-stranded DNA and / or cDNA. In some embodiments, “nucleic acid” encompasses ribonucleic acid (RNA), including but not limited to one or more of the following: interfering RNA (RNAi), small interfering RNA (siRNA), short hairpin RNA (shRNA), antisense RNA (aRNA), messenger RNA (mRNA), modified messenger RNA (mmRNA), long non-coding RNA (lncRNA), microRNA (miRNA), multimer-coding nucleic acid (MCNA), polymer-coated nucleic acid (PCNA), guide RNA (gRNA), and CRISPR RNA (crRNA). In some embodiments, “nucleic acid” encompasses deoxyribonucleic acid (DNA), including but not limited to one or more of the following: single-stranded DNA (ssDNA), double-stranded DNA (dsDNA), and complementary DNA (cDNA). In some embodiments, “nucleic acid” encompasses both RNA and DNA. In the embodiments, the DNA may be in the form of antisense DNA, plasmid DNA, a portion of plasmid DNA, pre-condensed DNA, polymerase chain reaction (PCR) products, vectors (e.g., P1, PAC, BAC, YAC, artificial chromosomes), expression cassettes, chimeric sequences, chromosomal DNA, or derivatives of these groups.In the embodiment, RNA includes messenger RNA (mRNA), ribosomal RNA (rRNA), signal recognition particle RNA (7SL RNA or SRP RNA), transcription RNA (tRNA), transcription messenger RNA (tmRNA), micronuclear RNA (snRNA), micronucleolar RNA (snoRNA), SmY RNA, small Cajal-specific RNA (scaRNA), guide RNA (gRNA), ribonuclease P (RNase P), Y RNA, telomerase RNA component (TERC), spliced ​​leader RNA (SL RNA), antisense RNA (aRNA or asRNA), cis-natural antisense transcript (cis-NAT), CRISPR RNA (crRNA), long non-coding RNA (lncRNA), microRNA (miRNA), piwi-interacting RNA (piRNA), small interfering RNA (siRNA), transaction siRNA (tasiRNA), repeat-associated siRNA (rasiRNA), and 73K It may be in the form of RNA, retrotransposons, viral genomes, viloids, satellite RNA, or derivatives of these groups. In some embodiments, the nucleic acid is mRNA that encodes a protein such as an enzyme.

[0161] Patient: As used herein, the terms “patient” or “subject” refer to any organism to which the provided composition may be administered, for example, for experimental, diagnostic, preventive, cosmetic, and / or therapeutic purposes. Typical patients include animals (e.g., mammals such as mice, rats, rabbits, non-human primates, and / or humans). In some embodiments, the patient is human. Humans include prenatal and postnatal forms.

[0162] Pharmaceutically acceptable: As used herein, the term "pharmaceutically acceptable" means a substance that, within reasonable medical judgment, is suitable for use in contact with human and animal tissues without excessive toxicity, irritation, allergic reactions, or other problems or complications, in proportion to a reasonable risk-benefit ratio.

[0163] Pharmaceutically acceptable salts: Pharmaceutically acceptable salts are well known in the art. For example, SMBerge et al. describe pharmaceutically acceptable salts in detail in J. Pharmaceutical Sciences (1977) 66:1-19. Pharmaceutically acceptable salts of the compounds of the present invention include those derived from suitable inorganic and organic acids and bases. Examples of pharmaceutically acceptable non-toxic acid addition salts are salts of amino groups formed with inorganic acids such as hydrochloric acid, hydrobromic acid, phosphoric acid, sulfuric acid, and perchloric acid, or with organic acids such as acetic acid, oxalic acid, maleic acid, tartaric acid, citric acid, succinic acid, or malonic acid, or by other methods used in the art, such as ion exchange. Other pharmaceutically acceptable salts include adipine, alginate, ascorbate, aspartate, benzenesulfonate, benzoate, bisulfate, borate, butyrate, camphorate, camphorsulfonate, citrate, cyclopentanepropionate, digluconate, dodecyl sulfate, ethanesulfonate, formate, fumarate, glucoheptonate, glycerophosphate, gluconate, hemisulfate, heptanoate, hexanoate, hydroiodide, and 2-hydroxyethanesulfonate. Examples include lactobionate, lactate, laurate, lauryl sulfate, malate, maleate, malonate, methanesulfonate, 2-naphthalenesulfonate, nicotinate, nitrate, oleate, oxalate, palmitate, pamoate, pectinate, persulfate, 3-phenylpropionate, phosphate, picrate, pivalate, propionate, stearate, succinate, sulfate, tartrate, thiocyanate, p-toluenesulfonate, undecanoate, and valerate. Salts derived from appropriate bases include alkali metal salts, alkaline earth metal salts, ammonium salts, and N + (C 1~4Examples include alkyl)4 salts. Typical alkali metal salts or alkaline earth metal salts include sodium, lithium, potassium, calcium, and magnesium. Further pharmaceutically acceptable salts include non-toxic ammonium, quaternary ammonium, and amine cations formed using counterions such as halides, hydroxides, carboxylates, sulfates, phosphates, nitrates, sulfonates, and arylsulfonates, where appropriate. Further pharmaceutically acceptable salts include salts formed from the quaternization of amines using electrophiles suitable for the formation of quaternized alkylated amino salts, such as alkyl halides.

[0164] Whole-body distribution or delivery: As used herein, the terms “whole-body distribution” or “whole-body delivery,” or their grammatical synonyms, refer to a mechanism or method of delivery or distribution that affects the entire body or organs. Typically, whole-body distribution or delivery is achieved through the body’s circulatory system, e.g., blood flow. Compare with the definition of “local distribution or delivery.”

[0165] Subject: As used herein, the term “subject” means a human or any non-human animal (e.g., mouse, rat, rabbit, dog, cat, cattle, pig, sheep, horse, or primate). Humans include prenatal and postnatal forms. In many embodiments, the subject is a human. The subject may be a patient, referring to a human being who visits a healthcare provider for the diagnosis or treatment of a disease. The term “subject” is used herein interchangeably with “individual” or “patient.” The subject may be a person suffering from or susceptible to a disease or disorder, the symptoms of which may or may not be present.

[0166] Substantially: As used herein, the term “substantially” refers to a qualitative condition indicating the overall or nearly entire range or degree of the desired feature or characteristic. Those skilled in the biological art will understand that biological and chemical phenomena, if any, are rarely completed and / or proceed completely, or achieve or avoid absolute results. Therefore, the term “substantially” is used herein to capture the inherent lack of integrity in many biological and chemical events.

[0167] Target tissue: As used herein, the term “target tissue” refers to any tissue affected by the disease to be treated. In some embodiments, target tissue includes tissue exhibiting a pathological condition, symptom, or characteristic associated with the disease.

[0168] Therapeutic dose: As used herein, the term “therapeutic dose” of a therapeutic agent means an amount sufficient to treat, diagnose, prevent and / or delay the onset of symptoms of a disease, disorder and / or condition when administered to a subject who is afflicted with or susceptible to such disease, disorder and / or condition. Those skilled in the art will understand that a therapeutic dose is typically administered in a dose-setting regimen comprising at least one unit dose.

[0169] Treatment: As used herein, the terms “treatment,” “therapy,” or “to treat” refer to any method used to partially or completely reduce, remit, alleviate, inhibit, prevent, delay the onset, reduce the severity, and / or reduce the incidence of one or more symptoms or characteristics of a particular disease, disorder, and / or condition. Treatment may be administered to subjects who are not showing signs of the disease and / or who are only showing early signs of the disease, for the purpose of reducing the risk of developing a disease-related pathological condition.

[0170] chemical definition Acyl: When used herein, the term "acyl" is R Z-(C=O)-(wherein, R Z This refers to, for example, any alkyl, alkenyl, alkynyl, heteroalkyl, or heteroalkylene.

[0171] Aliphatic: As used herein, the term aliphatic is C1-C 50 This refers to hydrocarbons, which include both saturated and unsaturated hydrocarbons. Aliphatic hydrocarbons can be linear, branched, or cyclic. For example, C1-C 20 Aliphatic cells are C1-C 20 Alkyl (e.g., linear or branched C1-C) 20 Saturated alkyl, C2~C 20 Alkenyls (e.g., linear or branched C4-C) 20 Dienyl, linear, or branched C6-C 20 (e.g., trienyl) and C2~C 20 Alkinyl (e.g., linear or branched C2-C) 20 (Alkinyl) may be included. C1~C 20 Aliphatic cells are C3-C 20 Cyclic aliphatic (e.g., C3~C) 20 Cycloalkyl, C4~C 20 Cycloalkenyl or C8~C 20 It may include cycloalkynyls. In certain embodiments, the aliphatic group may comprise one or more cyclic aliphatic and / or one or more heteroatoms (such as oxygen, nitrogen, or sulfur) and may optionally be substituted with one or more substituents such as alkyl, halo, alkoxy, hydroxy, amino, aryl, ether, ester, or amide. The aliphatic group may be unsubstituted or substituted with one or more substituents as described herein. For example, the aliphatic group may be substituted with one or more of halogens, -COR'', -CO2H, -CO2R'', -CN, -OH, -OR'', -OCOR', -OCO2R'', -NH2, -NHR'', -N(R'')2, -SR'', or -SO2R'' (e.g., 1, 2, 3, 4, 5, or 6 independently selected substituents), where each example of R'' is independently C1-C 20 Aliphatic (for example, C1-C 20 Alkyl, C1-C 15 Alkyl, C1-C 10It is an alkyl or C1-C3 alkyl. In the embodiment, R'' is independently an unsubstituted alkyl (e.g., unsubstituted C1-C3 alkyl). 20 Alkyl, C1-C 15 Alkyl, C1-C 10 It is alkyl or C1-C3 alkyl. In the embodiment, R'' is independently an unsubstituted C1-C3 alkyl. In the embodiment, the aliphatic is unsubstituted. In the embodiment, the aliphatic does not contain any heteroatoms.

[0172] Alkyl: As used herein, the term "alkyl" means acyclic straight-chain hydrocarbon groups and branched hydrocarbon groups, for example, "(C1~C 30 "Alkyl" refers to an alkyl group having 1 to 30 carbon atoms. Alkyl groups can be linear or branched. Examples of alkyl groups include, but are not limited to, methyl, ethyl, n-propyl, isopropyl, butyl, isobutyl, sec-butyl, tert-butyl, pentyl, isopentyl tert-pentylhexyl, and isohexyl. The term "lower alkyl" means a linear or branched alkyl group having 1 to 6 carbon atoms. Other alkyl groups will be readily apparent to those skilled in the art, given the advantages of this disclosure. Alkyl groups can be unsubstituted or substituted with one or more substituents, as described herein. For example, an alkyl group may be substituted with one or more of the following (e.g., 1, 2, 3, 4, 5, or 6 independently selected substituents): halogen, -COR'', -CO2H, -CO2R'', -CN, -OH, -OR'', -OCOR', -OCO2R'', -NH2, -NHR'', -N(R'')2, -SR'', or -SO2R'', where each example of R'' is independently C1-C 20 Aliphatic (for example, C1-C 20 Alkyl, C1-C 15 Alkyl, C1-C 10 It is an alkyl or C1-C3 alkyl. In the embodiment, R'' is independently an unsubstituted alkyl (e.g., unsubstituted C1-C3 alkyl). 20 Alkyl, C1-C 15 Alkyl, C1-C 10The alkyl group is either an alkyl group or a C1-C3 alkyl group. In embodiments, R'' is independently an unsubstituted C1-C3 alkyl group. In embodiments, the alkyl group is substituted (for example, with the 1, 2, 3, 4, 5, or 6 substituents described herein). In embodiments, the alkyl group is substituted with an -OH group, which may also be referred to herein as a "hydroxyalkyl" group, where the prefix represents the -OH group and "alkyl" is as described herein.

[0173] As used herein, “alkyl” also refers to a linear or branched saturated hydrocarbon group having 1 to 50 carbon atoms ("C1-C50"). 50 Alkyl). In some embodiments, alkyl groups have 1 to 40 carbon atoms ("C1-C"). 40 Alkyl). In some embodiments, the alkyl group has 1 to 30 carbon atoms ("C1-C30"). 30 Alkyl). In some embodiments, the alkyl group has 1 to 20 carbon atoms ("C1-C20"). 20 Alkyl). In some embodiments, the alkyl group has 1 to 10 carbon atoms ("C1-C10"). 101. C1-C9 alkyl group. 2. C1 alkyl group. 3. C1 alkyl group. 4. C1 alkyl group. 5. C1 alkyl group. 6. C1 alkyl group. 7. C1 alkyl group. 8. C1 alkyl group. 9. C1 alkyl group. 1 Examples of C1-C6 alkyl groups include, but are not limited to, methyl (C1), ethyl (C2), n-propyl (C3), isopropyl (C3), n-butyl (C4), tert-butyl (C4), sec-butyl (C4), isobutyl (C4), n-pentyl (C5), 3-pentanyl (C5), amyl (C5), neopentyl (C5), 3-methyl-2-butanyl (C5), tertiary amyl (C5), and n-hexyl (C6). Further examples of alkyl groups include n-heptyl (C7) and n-octyl (C8). Unless otherwise specified, each example of an alkyl group is independently either unsubstituted ("unsubstituted alkyl") or substituted with one or more substituents ("substituted alkyl"). In certain embodiments, the alkyl group is unsubstituted C1-C 50 It is alkyl. In certain embodiments, the alkyl group is substituted C1-C 50 It is alkyl.

[0174] Adding the suffix "-en" to a base indicates that the base is a divalent part; for example, arylene is the divalent part of aryl, and heteroarylene is the divalent part of heteroaryl.

[0175] Alkylene: As used herein, the term "alkylene" refers to a saturated divalent linear or branched hydrocarbon group, exemplified by methylene, ethylene, isopropylene, and the like. Similarly, as used herein, the term "alkenylene" refers to an unsaturated divalent linear or branched hydrocarbon group having one or more unsaturated carbon-carbon double bonds that may occur at any stable point along the chain, and as used herein, the term "alkynylene" refers to an unsaturated divalent linear or branched hydrocarbon group having one or more unsaturated carbon-carbon triple bonds that may occur at any stable point along the chain. In certain embodiments, the alkylene, alkenylene, or alkynylene group may contain one or more cyclic aliphatic and / or one or more heteroatoms (such as oxygen, nitrogen, or sulfur) and may optionally be substituted with one or more substituents such as alkyl, halo, alkoxy, hydroxy, amino, aryl, ether, ester, or amide. For example, alkylene, alkenylene, or alkynylene may be substituted with one or more of the following substituents (e.g., 1, 2, 3, 4, 5, or 6 independently selected substituents): halogen, -COR'', -CO2H, -CO2R'', -CN, -OH, -OR'', -OCOR'', -OCO2R'', -NH2, -NHR'', -N(R'')2, -SR'', or -SO2R'', where each instance of R'' is independently C1-C 20 Aliphatic (for example, C1-C 20 Alkyl, C1-C 15 Alkyl, C1-C 10 It is an alkyl or C1-C3 alkyl. In the embodiment, R'' is independently an unsubstituted alkyl (e.g., unsubstituted C1-C3 alkyl). 20 Alkyl, C1-C 15 Alkyl, C1-C 10 It is alkyl or C1-C3 alkyl. In the embodiment, R'' is independently an unsubstituted C1-C3 alkyl. In certain embodiments, alkylene, alkenylene or alkynylene is unsubstituted. In certain embodiments, alkylene, alkenylene or alkynylene does not contain any heteroatoms.

[0176] Alkenyl: As used herein, “alkenyl” means any straight or branched hydrocarbon chain having one or more unsaturated carbon-carbon double bonds that can occur at any stable point along the chain, for example, “(C2~C 30 "Alkenyl" refers to an alkenyl group having 2 to 30 carbon atoms. For example, alkenyl groups include propa-2-enyl, buta-2-enyl, buta-3-enyl, 2-methylpropa-2-enyl, hexa-2-enyl, hexa-5-enyl, and 2,3-dimethylbuta-2-enyl. In embodiments, the alkenyl contains 1, 2, or 3 carbon-carbon double bonds. In embodiments, the alkenyl contains a single carbon-carbon double bond. In embodiments, multiple double bonds (e.g., 2 or 3) are conjugated. The alkenyl group may be unsubstituted or substituted with one or more substituents as described herein. For example, the alkenyl group may be substituted with one or more of the following substituents (e.g., 1, 2, 3, 4, 5, or 6 independently selected substituents): halogen, -COR'', -CO2H, -CO2R'', -CN, -OH, -OR'', -OCOR'', -OCO2R'', -NH2, -NHR'', -N(R'')2, -SR'', or -SO2R'', where each example of R'' is independently C1-C 20 Aliphatic (for example, C1-C 20 Alkyl, C1-C 15 Alkyl, C1-C 10 It is an alkyl or C1-C3 alkyl. In the embodiment, R'' is independently an unsubstituted alkyl (e.g., unsubstituted C1-C3 alkyl). 20 Alkyl, C1-C 15 Alkyl, C1-C 10 The group is alkyl or C1-C3 alkyl. In embodiments, R'' is independently an unsubstituted C1-C3 alkyl. In embodiments, the alkenyl is unsubstituted. In embodiments, the alkenyl is substituted (for example, with the 1, 2, 3, 4, 5, or 6 substituents described herein). In embodiments, the alkenyl group is substituted with an -OH group, which may also be referred to herein as a "hydroxyalkenyl" group, where the prefix represents the -OH group and "alkenyl" is as described herein.

[0177] As used herein, “alkenyl” also refers to a linear or branched hydrocarbon group having 2 to 50 carbon atoms and one or more carbon-carbon double bonds (e.g., 1, 2, 3, or 4 double bonds) (“C2~C 50 "Alkenyl"). In some embodiments, the alkenyl group has 2 to 40 carbon atoms ("C2-C2"). 40 ("Alkenyl"). In some embodiments, the alkenyl group has 2 to 30 carbon atoms ("C2-C30"). 30 ("Alkenyl"). In some embodiments, the alkenyl group has 2 to 20 carbon atoms ("C2-C2"). 20 ("Alkenyl"). In some embodiments, the alkenyl group has 2 to 10 carbon atoms ("C2-C10"). 10Alkenyl). In some embodiments, the alkenyl group has 2 to 9 carbon atoms ("C2-C9 alkenyl"). In some embodiments, the alkenyl group has 2 to 8 carbon atoms ("C2-C8 alkenyl"). In some embodiments, the alkenyl group has 2 to 7 carbon atoms ("C2-C7 alkenyl"). In some embodiments, the alkenyl group has 2 to 6 carbon atoms ("C2-C6 alkenyl"). In some embodiments, the alkenyl group has 2 to 5 carbon atoms ("C2-C5 alkenyl"). In some embodiments, the alkenyl group has 2 to 4 carbon atoms ("C2-C4 alkenyl"). In some embodiments, the alkenyl group has 2 to 3 carbon atoms ("C2-C3 alkenyl"). In some embodiments, the alkenyl group has 2 carbon atoms ("C2 alkenyl"). One or more carbon-carbon double bonds may be internal (e.g., in 2-butenyl) or terminal (e.g., in 1-butenyl). Examples of C2-C4 alkenyl groups include, but are not limited to, ethenyl (C2), 1-propenyl (C3), 2-propenyl (C3), 1-butenyl (C4), 2-butenyl (C4), and butadienyl (C4). Examples of C2-C6 alkenyl groups include the aforementioned C2-C4 alkenyl groups, as well as pentenyl (C5), pentadienyl (C5), and hexenyl (C6). Further examples of alkenyls include heptenyl (C7), octenyl (C8), and octatrienyl (C8). Unless otherwise specified, each example of an alkenyl group is independently either unsubstituted ("unsubstituted alkenyl") or substituted with one or more substituents ("substituted alkenyl"). In certain embodiments, the alkenyl group is an unsubstituted C2-C 50 It is an alkenyl. In certain embodiments, the alkenyl group is a substituted C2-C 50 It is Alkenil.

[0178] Alkynyl: As used herein, "alkynyl" means any straight or branched hydrocarbon chain having one or more carbon-carbon triple bonds at any stable point along the chain, for example, "C2-C 30"Alkynyl" refers to an alkynyl group having 2 to 30 carbon atoms. Examples of alkynyl groups include propa-2-inyl, buta-2-inyl, buta-3-inyl, penta-2-inyl, 3-methylpenta-4-inyl, hexa-2-inyl, and hexa-5-inyl. In embodiments, the alkynyl contains one carbon-carbon triple bond. The alkynyl group may be unsubstituted or substituted with one or more substituents as described herein. For example, the alkynyl group may be substituted with one or more of the following (e.g., 1, 2, 3, 4, 5, or 6 independently selected substituents): halogen, -COR'', -CO2H, -CO2R'', -CN, -OH, -OR'', -OCOR'', -OCO2R'', -NH2, -NHR'', -N(R'')2, -SR'', or -SO2R'', where each example of R'' is independently C1-C 20 Aliphatic (for example, C1-C 20 Alkyl, C1-C 15 Alkyl, C1-C 10 It is an alkyl or C1-C3 alkyl. In the embodiment, R'' is independently an unsubstituted alkyl (e.g., unsubstituted C1-C3 alkyl). 20 Alkyl, C1-C 15 Alkyl, C1-C 10 The alkyl group is an alkyl group or a C1-C3 alkyl group. In embodiments, R'' is independently an unsubstituted C1-C3 alkyl group. In embodiments, the alkynyl group is unsubstituted. In embodiments, the alkynyl group is substituted (for example, with the 1, 2, 3, 4, 5, or 6 substituents described herein).

[0179] As used herein, "alkynyl" also refers to a linear or branched hydrocarbon group having 2 to 50 carbon atoms and one or more carbon-carbon triple bonds (e.g., 1, 2, 3, or 4 triple bonds) and optionally one or more double bonds (e.g., 1, 2, 3, or 4 double bonds) ("C2-C 50 Alkynyl groups, which have one or more triple bonds and one or more double bonds, are also called "en-yne" groups. In some embodiments, the alkynyl group has 2 to 40 carbon atoms ("C2-C2"). 40In some embodiments, the alkynyl group has 2 to 30 carbon atoms ("C2-C30"). 30 In some embodiments, the alkynyl group has 2 to 20 carbon atoms ("C2-C2"). 20 In some embodiments, the alkynyl group has 2 to 10 carbon atoms ("C2-C10"). 10 In some embodiments, the alkynyl group has 2 to 9 carbon atoms ("C2-C9 alkynyl"). In some embodiments, the alkynyl group has 2 to 8 carbon atoms ("C2-C8 alkynyl"). In some embodiments, the alkynyl group has 2 to 7 carbon atoms ("C2-C7 alkynyl"). In some embodiments, the alkynyl group has 2 to 6 carbon atoms ("C2-C6 alkynyl"). In some embodiments, the alkynyl group has 2 to 5 carbon atoms ("C2-C5 alkynyl"). In some embodiments, the alkynyl group has 2 to 4 carbon atoms ("C2-C4 alkynyl"). In some embodiments, the alkynyl group has 2 to 3 carbon atoms ("C2-C3 alkynyl"). In some embodiments, the alkynyl group has 2 carbon atoms ("C2 alkynyl"). One or more carbon-carbon triple bonds may be internal (e.g., in 2-butynyl) or terminal (e.g., in 1-butynyl). Examples of C2-C4 alkynyl groups include, but are not limited to, ethynyl (C2), 1-propynyl (C3), 2-propynyl (C3), 1-butynyl (C4), and 2-butynyl (C4). Examples of C2-C6 alkenyl groups include the aforementioned C2-C4 alkynyl groups as well as pentynyl (C5) and hexynyl (C6). Further examples of alkynyls include heptynyl (C7) and octinyl (C8). Unless otherwise specified, each example of an alkynyl group is independently unsubstituted ("unsubstituted alkynyl") or substituted with one or more substituents ("substituted alkynyl"). In certain embodiments, the alkynyl group is unsubstituted C2-C 50 It is an alkynyl group. In certain embodiments, the alkynyl group is a substituted C2-C 50 It is alkinyl.

[0180] Aryl: Used alone or as part of a larger term such as “aralkyl”, the term “aryl” refers to a monocyclic, bicyclic, or tricyclic carbocyclic system having a total of 6 to 14 ring members, wherein the system has a single bond to the rest of the molecule, at least one ring in the system is aromatic, and each ring in the system contains 4 to 7 ring members. In some embodiments, the aryl group has 6 ring carbon atoms ("(C6)aryl", e.g., phenyl). In some embodiments, the aryl group has 10 ring carbon atoms ("C6"). 10 "Aryl" (for example, naphthyl such as 1-naphthyl and 2-naphthyl). In some embodiments, the aryl group has 14 ring carbon atoms ("C"). 14 "Aryl" (e.g., anthracyl). "Aryl" also includes ring systems in which the aryl ring defined above is condensed with one or more carbocyrillic or heterocyclyl groups, and the radical or bond site is on the aryl ring, in which case the number of carbon atoms continues to specify the number of carbon atoms in the aryl ring system. Exemplary aryls include phenyl, naphthyl, and anthracene.

[0181] As used herein, “aryl” also refers to monocyclic or polycyclic (e.g., bicyclic or tricyclic) 4n+2 aromatic ring systems (e.g., having 6, 10, or 14 shared π electrons within a cyclic arrangement) in which the aromatic ring system is provided with 6 to 14 ring carbon atoms and 0 heteroatoms (“C6~C 14 In some embodiments, the aryl group has six ring carbon atoms ("C6 aryl"; e.g., phenyl). In some embodiments, the aryl group has ten ring carbon atoms ("C6 aryl"). 10 "Aryl"; for example, naphthyl (such as 1-naphthyl and 2-naphthyl). In some embodiments, the aryl group has 14 ring carbon atoms ("C"). 14"Aryl" (e.g., anthracyl). "Aryl" also includes ring systems in which the aryl ring as defined above is condensed with one or more carbocyrillic or heterocyclyl groups, and the radical or bond site is on the aryl ring, in which case the number of carbon atoms continues to specify the number of carbon atoms in the aryl ring system. Unless otherwise specified, each example of an aryl group is independently unsubstituted ("unsubstituted aryl") or substituted with one or more substituents ("substituted aryl"). In certain embodiments, the aryl group is unsubstituted C6-C 14 It is aryl. In certain embodiments, the aryl group is substituted C6-C 14 It is Ariel.

[0182] Arylene: As used herein, the term "arylene" refers to a divalent (i.e., a molecule having two bonding sites) aryl group. Exemplary arylenes include phenylene (e.g., unsubstituted phenylene or substituted phenylene).

[0183] Carbocyclyl: As used herein, "carbocyclyl" or "carbocyclic" refers to a non-aromatic ring system with 3 to 10 ring carbon atoms ("C3-C3"). 10 This refers to a group of non-aromatic cyclic hydrocarbon groups having 0 heteroatoms ("C3-C8 carbocyclyl"). In some embodiments, the carbocyclyl group has 3-8 ring carbon atoms ("C3-C8 carbocyclyl"). In some embodiments, the carbocyclyl group has 3-7 ring carbon atoms ("C3-C7 carbocyclyl"). In some embodiments, the carbocyclyl group has 3-6 ring carbon atoms ("C3-C6 carbocyclyl"). In some embodiments, the carbocyclyl group has 4-6 ring carbon atoms ("C4-C6 carbocyclyl"). In some embodiments, the carbocyclyl group has 5-6 ring carbon atoms ("C5-C6 carbocyclyl"). In some embodiments, the carbocyclyl group has 5-10 ring carbon atoms ("C5-C6 carbocyclyl"). 10Examples of C3-C6 carbocyclyl groups include, but are not limited to, cyclopropyl (C3), cyclopropenyl (C3), cyclobutyl (C4), cyclobutenyl (C4), cyclopentyl (C5), cyclopentenyl (C5), cyclohexyl (C6), cyclohexenyl (C6), and cyclohexadienyl (C6). Examples of C3-C8 carbocyclyl groups include, but are not limited to, the aforementioned C3-C6 carbocyclyl groups, as well as cycloheptyl (C7), cycloheptenyl (C7), cycloheptadienyl (C7), cycloheptatrienyl (C7), cyclooctyl (C8), cyclooctenyl (C8), bicyclo[2.2.1]heptanyl (C7), and bicyclo[2.2.2]octanyl (C8). Exemplary C3~C 10 The carbocyclyl group is not limited to the aforementioned C3-C8 carbocyclyl group, as well as cyclononyl (C9), cyclononenyl (C9), and cyclodecyl (C9). 10 ), cyclodecenyl (C 10 ), octahydro-1H-indenyl (C9), decahydronaphthalenyl (C9) 10 ), spiro[4.5]decanil(C 10 ) are some examples. As the above examples show, in certain embodiments, the carbocyclyl group may be monocyclic ("monocyclic carbocyclyl") or polycyclic (e.g., bicyclic ("bicyclic carbocyclyl") or tricyclic (including condensed, bridged, or spirocyclic systems such as "tricyclic carbocyclyl")), and may be saturated or contain one or more carbon-carbon double or triple bonds. "Carbocyclyl" also includes cyclic systems in which the carbocyclyl ring as defined above is condensed with one or more aryl or heteroaryl groups, and the radical or bond site is on the carbocyclyl ring, in which case the number of carbon atoms continues to specify the number of carbon atoms in the carbocyclyl cyclic system. Unless otherwise specified, each example of a carbocyclyl group is independently unsubstituted ("unsubstituted carbocyclyl") or substituted with one or more substituents ("substituted carbocyclyl"). In certain embodiments, the carbocyclyl group may be unsubstituted (C3-C 10 ) is a carbocyclyl. In certain embodiments, the carbocyclyl group is substituted C3~C10 It is carbocyclyl.

[0184] In some embodiments, "carbocyrill" or "carbocyclic" is referred to as "cycloalkyl," that is, a monocyclic saturated carbocyclyl group having 3 to 10 ring carbon atoms ("C3-C 10 In some embodiments, the cycloalkyl group has 3 to 8 ring carbon atoms ("C3-C8 cycloalkyl"). In some embodiments, the cycloalkyl group has 3 to 6 ring carbon atoms ("C3-C6 cycloalkyl"). In some embodiments, the cycloalkyl group has 4 to 6 ring carbon atoms ("C4-C6 cycloalkyl"). In some embodiments, the cycloalkyl group has 5 to 6 ring carbon atoms ("C5-C6 cycloalkyl"). In some embodiments, the cycloalkyl group has 5 to 10 ring carbon atoms ("C5-C6 cycloalkyl"). 10 (Cycloalkyl). Examples of C5-C6 cycloalkyl groups include cyclopentyl (C5) and cyclohexyl (C5). Examples of C3-C6 cycloalkyl groups include the aforementioned C5-C6 cycloalkyl groups as well as cyclopropyl (C3) and cyclobutyl (C4). Examples of C3-C8 cycloalkyl groups include the aforementioned C3-C6 cycloalkyl groups as well as cycloheptyl (C7) and cyclooctyl (C8). Unless otherwise specified, each example of a cycloalkyl group is either unsubstituted ("unsubstituted cycloalkyl") or substituted with one or more substituents ("substituted cycloalkyl"). In certain embodiments, the cycloalkyl group is unsubstituted C3-C 10 It is a cycloalkyl group. In certain embodiments, the cycloalkyl group is a substituted C3-C3 group. 10 It is a cycloalkyl group.

[0185] Halogen: As used herein, the term "halogen" means fluorine, chlorine, bromine, or iodine.

[0186] Heteroalkyl: The term "heteroalkyl" means a branched or unbranched alkyl, alkenyl, or alkynyl group having 4 to 50 carbon atoms in addition to 1, 2, 3, or 4 heteroatoms independently selected from the group consisting of N, O, S, and P. In embodiments, the heteroalkyl group has 4 to 40 carbon atoms ("C4-C4"). 40 ("heteroalkyl"). In some embodiments, the heteroalkyl has 6 to 30 carbon atoms ("C6-C30"). 30 ("heteroalkyl"). In some embodiments, the heteroalkyl group has 4 to 20 carbon atoms ("C4-C20"). 20 Heteroalkyl groups (e.g., C4-C4 alkyl groups). Examples of heteroalkyl groups include tertiary amines, secondary amines, ethers, esters, thioethers, amides, thioamides, carbamates, thiocarbamates, hydrazones, imines, phosphodiesters, phosphoramidates, sulfonamides, and disulfides. Heteroalkyl groups may optionally include monocyclic, bicyclic, or tricyclic rings, each ring preferably having 3 to 6 ring members. Examples of heteroalkyl groups include polyethers such as methoxymethyl and ethoxyethyl. In embodiments, heteroalkyl groups may be unsubstituted or substituted (e.g., including hydroxyl groups, oxo groups, or ionizable nitrogen groups as described herein) or alkyl groups as described herein (e.g., C4-C4 50 Alkyl or C6-C 30 It comprises an alkyl group and / or one or two heteroatoms selected from N and O. In embodiments, the heteroalkyl group comprises an oxo substituent (for example, the heteroalkyl group comprises an ester group in which the oxo is adjacent to an oxygen atom).

[0187] Heteroalkylene: When used herein, the term "heteroalkylene" refers to the divalent form of the heteroalkyl group described herein.

[0188] Heteroaryl: The term "heteroaryl," as used herein, refers to a fully unsaturated heteroatom-containing ring in which at least one ring atom is a heteroatom such as nitrogen and oxygen, but is not limited to these.

[0189] As used herein, “heteroaryl” also refers to a 5- to 14-membered monocyclic or polycyclic (e.g., bicyclic or tricyclic) 4n+2 aromatic ring system (e.g., having 6, 10 or 14 π electrons shared in a cyclic arrangement) radical having a ring carbon atom and one or more ring heteroatoms (e.g., 1, 2, 3, or 4 ring heteroatoms) in an aromatic ring system, where each heteroatom is independently selected from oxygen, sulfur, nitrogen, boron, silicon, and phosphorus ("5- to 14-membered heteroaryl"). In heteroaryl groups containing one or more nitrogen atoms, the bond site can be a carbon atom or a nitrogen atom, as long as the valence allows. Heteroaryl polycyclic ring systems may contain one or more heteroatoms in one or both rings. “Heteroaryl” includes ring systems in which the heteroaryl ring as defined above is condensed with one or more carbocyryl or heterocyclyl groups, and the bond site is on the heteroaryl ring, in which case the number of ring members continues to specify the number of ring members in the heteroaryl ring system. "Hyperaryl" includes ring systems in which a heteroaryl ring as defined above is fused with one or more aryl groups, and the bond site is on an aryl or heteroaryl ring, in which case the number of ring members specifies the number of ring members in the fused polycyclic (aryl / heteroaryl) ring system. A polycyclic heteroaryl group in which one ring does not contain a heteroatom (e.g., indolyl, quinolinyl, carbazolyl, etc.) may have a bond site on either ring, i.e., a ring with a heteroatom (e.g., 2-indolyl) or a ring without a heteroatom (e.g., 5-indolyl).

[0190] In some embodiments, the heteroaryl group is a 5- to 10-membered aromatic ring system having a ring carbon atom provided to the aromatic ring system and one or more (e.g., 1, 2, 3, or 4) ring heteroatoms, where each heteroatom is independently selected from oxygen, sulfur, nitrogen, boron, silicon, and phosphorus ("5- to 10-membered heteroaryl"). In some embodiments, the heteroaryl group is a 5- to 8-membered aromatic ring system having a ring carbon atom provided to the aromatic ring system and one or more (e.g., 1, 2, 3, or 4) ring heteroatoms, where each heteroatom is independently selected from oxygen, sulfur, nitrogen, boron, silicon, and phosphorus ("5- to 8-membered heteroaryl"). In some embodiments, the heteroaryl group is a 5-6 membered aromatic ring system having a ring carbon atom provided to the aromatic ring system and one or more (e.g., 1, 2, 3, or 4) ring heteroatoms, where each heteroatom is independently selected from oxygen, sulfur, nitrogen, boron, silicon, and phosphorus ("5-6 membered heteroaryl"). In some embodiments, the 5-6 membered heteroaryl has one or more (e.g., 1, 2, or 3) ring heteroatoms selected from oxygen, sulfur, nitrogen, boron, silicon, and phosphorus. In some embodiments, the 5-6 membered heteroaryl has one or two ring heteroatoms selected from oxygen, sulfur, nitrogen, boron, silicon, and phosphorus. In some embodiments, the 5-6 membered heteroaryl has one ring heteroatom selected from oxygen, sulfur, nitrogen, boron, silicon, and phosphorus. Unless otherwise specified, each example of a heteroaryl group is independently unsubstituted ("unsubstituted heteroaryl") or substituted with one or more substituents ("substituted heteroaryl"). In certain embodiments, the heteroaryl group is an unsubstituted 5- to 14-membered heteroaryl group.

[0191] Examples of five-membered heteroaryl groups containing one heteroatom include, but are not limited to, pyrrolyl, furanyl, and thiophenyl. Examples of five-membered heteroaryl groups containing two heteroatoms include, but are not limited to, imidazolyl, pyrazolyl, oxazolyl, isoxazolyl, thiazolyl, and isothiazolyl. Examples of five-membered heteroaryl groups containing three heteroatoms include, but are not limited to, triazolyl, oxadiazolyl, and thiadiazolyl. Examples of five-membered heteroaryl groups containing four heteroatoms include, but are not limited to, tetrazolyl. Examples of six-membered heteroaryl groups containing one heteroatom include, but are not limited to, pyridinyl. Examples of six-membered heteroaryl groups containing two heteroatoms include, but are not limited to, pyridazinyl, pyrimidinyl, and pyrazinyl. Examples of six-membered heteroaryl groups containing three or four heteroatoms include, but are not limited to, triazinyl and tetradinyl, respectively. Examples of seven-membered heteroaryl groups containing one heteroatom include, but are not limited to, azepinyl, oxepinyl, and thiepinyl. Examples of 5,6-bicyclic heteroaryl groups include, but are not limited to, indolyl, isoindolyl, indazolyl, benzotriazolyl, benzothiophenyl, isobenzothiophenyl, benzofuranil, benzoisofuranil, benzimidazolyl, benzoxazolyl, benzoisoxazolyl, benzoxadiazolyl, benzothiazolyl, benzoisothiazolyl, benzothiadiazolyl, indolidinyl, and purinyl. Examples of 6,6-bicyclic heteroaryl groups include, but are not limited to, naphthylidinyl, pteridinyl, quinolinyl, isoquinolinyl, sinnolinyl, quinoxalinyl, phthalazinyl, and quinazolinyl. Examples of exemplary tricyclic heteroaryl groups include, but are not limited to, phenanthridine, dibenzofuranil, carbazolyl, acridinil, phenothiazinil, phenoxadinil, and phenadinil.

[0192] As used herein, “heterocyclyl” or “heterocyclic” refers to a radical of a 3- to 14-membered non-aromatic ring system having a ring carbon atom and one or more (e.g., 1, 2, 3, or 4) ring heteroatoms, each heteroatom independently selected from oxygen, sulfur, nitrogen, boron, silicon, and phosphorus (“3- to 14-membered heterocyclyl”). In heterocyclyl groups containing one or more nitrogen atoms, the bond sites may be carbon atoms or nitrogen atoms, as long as the valence allows. Heterocyclyl groups may be monocyclic (“monocyclic heterocyclyl”) or polycyclic (e.g., bicyclic (“bicyclic heterocyclyl”) or tricyclic (“tricyclic heterocyclyl”), condensed, bridging, or spirocyclic systems), and may be saturated or contain one or more carbon-carbon double or triple bonds. Heterocyclyl polycyclic ring systems may contain one or more heteroatoms in one or both rings. "Heterocyclyl" also includes ring systems in which a heterocyclyl ring as defined above is fused with one or more carbocykyl groups, with the bonding site being either a carbocykyl ring or a heterocyclyl ring, or ring systems in which a heterocyclyl ring as defined above is fused with one or more aryl or heteroaryl groups, with the bonding site being a heterocyclyl ring, and in such cases, the number of ring members refers to the number of ring members that remain within the heterocyclyl ring system. Unless otherwise specified, each example of a heterocyclyl group is independently unsubstituted ("unsubstituted heterocyclyl") or substituted with one or more substituents ("substituted heterocyclyl"). In certain embodiments, the heterocyclyl group is an unsubstituted 3- to 14-membered heterocyclyl.

[0193] In some embodiments, the heterocyclyl group is a 5-10 member non-aromatic ring system having a ring carbon atom and one or more (e.g., 1, 2, 3, or 4) ring heteroatoms, where each heteroatom is independently selected from oxygen, sulfur, nitrogen, boron, silicon, and phosphorus ("5-10 member heterocyclyl"). In some embodiments, the heterocyclyl group is a 5-8 member non-aromatic ring system having a ring carbon atom and one or more (e.g., 1, 2, 3, or 4) ring heteroatoms, where each heteroatom is independently selected from oxygen, sulfur, nitrogen, boron, silicon, and phosphorus ("5-8 member heterocyclyl"). In some embodiments, the heterocyclyl group is a 5-6 member non-aromatic ring system having a ring carbon atom and one or more (e.g., 1, 2, 3, or 4) ring heteroatoms, where each heteroatom is independently selected from oxygen, sulfur, nitrogen, boron, silicon, and phosphorus ("5-6 member heterocyclyl"). In some embodiments, the 5-6 membered heterocyclil has one or more (e.g., 1, 2, or 3) ring heteroatoms selected from oxygen, sulfur, nitrogen, boron, silicon, and phosphorus. In some embodiments, the 5-6 membered heterocyclil has one or two ring heteroatoms selected from oxygen, sulfur, nitrogen, boron, silicon, and phosphorus. In some embodiments, the 5-6 membered heterocyclil has one ring heteroatom selected from oxygen, sulfur, nitrogen, boron, silicon, and phosphorus.

[0194] Examples of three-membered heterocyclyl groups having one heteroatom include, but are not limited to, azirdinyl, oxylanil, and thiorenyl. Examples of four-membered heterocyclyl groups having one heteroatom include, but are not limited to, azetidinyl, oxetanil, and thietanil. Examples of five-membered heterocyclyl groups having one heteroatom include, but are not limited to, tetrahydrofuranil, dihydrofuranil, tetrahydrothiophenyl, dihydrothiophenyl, pyrrolidinyl, dihydropyrrolyl, and pyrrolyl-2,5-dione. Examples of five-membered heterocyclyl groups having two heteroatoms include, but are not limited to, dioxolanil, oxathiolanil, and dithiolanil. Examples of five-membered heterocyclyl groups having three heteroatoms include, but are not limited to, triazolinyl, oxadiazolinyl, and thiadiazolinyl. Examples of six-membered heterocyclyl groups having one heteroatom include, but are not limited to, piperidinyl, tetrahydropyranil, dihydropyridinyl, and thianil. Examples of six-membered heterocyclyl groups having two heteroatoms include, but are not limited to, piperazinyl, morpholinil, dithianil, and dioxanil. Examples of six-membered heterocyclyl groups having two heteroatoms include, but are not limited to, triazinyl. Examples of seven-membered heterocyclyl groups having one heteroatom include, but are not limited to, azepanil, oxepanil, and thiepanil. Examples of eight-membered heterocyclyl groups having one heteroatom include, but are not limited to, azokanil, oxecanil, and thiokanil.Examples of bicyclic heterocyclyl groups include, but are not limited to, indolinyl, isoindolinyl, dihydrobenzofuranyl, dihydrobenzothienyl, tetrahydrobenzothienyl, tetrahydrobenzofuranyl, tetrahydroindolyl, tetrahydroquinolinyl, tetrahydroisoquinolinyl, decahydroquinolinyl, decahydroisoquinolinyl, octahydroclomenyl, octahydroisoclomenyl, decahydronaphthilidinyl, decahydro-1,8-naphthilidinyl, octahydropyrrolo[3,2-b]pyrrole, indolinyl, phthaliumidyl, naphthaliumidyl, chromanyl, clomenyl, and 1H-benzo[e][1,4]diazepi Examples include nyl, 1,4,5,7-tetrahydropyrano[3,4-b]pyrrolyl, 5,6-dihydro-4H-fl[3,2-b]pyrrolyl, 6,7-dihydro-5H-fl[3,2-b]pyranyl, 5,7-dihydro-4H-thieno[2,3-c]pyranyl, 2,3-dihydro-1H-pyrrolo[2,3-b]pyridinyl, 2,3-dihydrofl[2,3-b]pyridinyl, 4,5,6,7-tetrahydro-1H-pyrrolo-[2,3-b]pyridinyl, 4,5,6,7-tetrahydrofl[3,2-c]pyridinyl, 4,5,6,7-tetrahydrothieno[3,2-b]pyridinyl, and 1,2,3,4-tetrahydro-1,6-naphthilidinyl.

[0195] Heterocycloalkyl: As used herein, the term "heterocycloalkyl" refers to a non-aromatic ring in which at least one atom is a heteroatom such as nitrogen, oxygen, sulfur, or phosphorus, and the remaining atoms are carbon. Heterocycloalkyl groups may be substituted or unsubstituted.

[0196] As can be understood from the above, the alkyl, alkenyl, alkynyl, acyl, carbocyclyl, heterocyclyl, aryl, and heteroaryl groups as defined herein are optionally substituted in certain embodiments. Optionally substituted means a group that may be substituted or unsubstituted (e.g., a "substituted" or "unsubstituted" alkyl, a "substituted" or "unsubstituted" alkenyl, a "substituted" or "unsubstituted" alkynyl, a "substituted" or "unsubstituted" heteroalkyl, a "substituted" or "unsubstituted" heteroalkenyl, a "substituted" or "unsubstituted" heteroalkynyl, a "substituted" or "unsubstituted" carbocyclyl, a "substituted" or "unsubstituted" heterocyclyl, a "substituted" or "unsubstituted" aryl, or a "substituted" or "unsubstituted" heteroaryl group). Generally, the term "substituted" means that at least one hydrogen present on the group does not spontaneously undergo transformation by an acceptable substituent, such as substitution, resulting in a stable compound, such as reconstitution, cyclization, removal, or other reactions. This means that the group is substituted with a substituent. Unless otherwise indicated, a “substituted” group has substituents at one or more substituted positions of the group, and if two or more positions in any given structure are substituted, the substituents are either the same or different at each position. The term “substituted” is intended to include substitution with any of the substituents described herein that result in the formation of a stable compound, among all acceptable substituents of an organic compound. The present invention intends any and all such combinations to arrive at a stable compound. For the purposes of the present invention, heteroatoms such as nitrogen may have hydrogen substituents and / or any preferred substituents as described herein that satisfy the valence of the heteroatom and result in the formation of a stable moiety.

[0197] Examples of carbon atom substituents include, but are not limited to, halogens, -CN, -NO2, -N3, -SO2, -SO3H, -OH, and -OR aa , -ON(R bb )2, -N(R bb )2, -N(R bb )3+X-, -N(OR cc )R bb -SeH, -SeR aa、-SH、-SR aa 、-SSR cc 、-C(=O)R aa 、-CO2H、-CHO、-C(OR cc )2、-CO2R aa 、-OC(=O)R aa 、-OCO2R aa 、-C(=O)N(R bb )2、-OC(=O)N(R bb )2、-NR bb C(=O)R aa 、-NR bb CO2R aa 、-NR bb C(=O)N(R bb )2、-C(=NR bb )R aa 、-C(=NR bb )OR aa 、-OC(=NR bb )R aa 、-OC(=NR bb )OR aa 、-C(=NR bb )N(R bb )2、-OC(=NR bb )N(R bb )2、-NR bb C(=NR bb )N(R bb )2、-C(=O)NR bb SO2R aa 、-NR bb SO2R aa 、-SO2N(R bb )2、-SO2R aa 、-SO2OR aa 、-OSO2R aa 、-S(=O)R aa 、-OS(=O)R aa 、-Si(R aa )3-OSi(R aa )3-C(=S)N(R bb )2、-C(=O)SR aa 、-C(=S)SR aa 、-SC(=S)SR aa 、-SC(=O)SR aa 、-OC(=O)SR aa 、-SC(=O)OR aa 、-SC(=O)Raa -P(=O)2R aa -OP(=O)2R aa -P(=O)(R aa )2, -OP(=O)(R aa )2, -OP(=O)(OR cc )2, -P(=O)2N(R bb )2, -OP(=O)2N(R bb )2, -P(=O)(NR bb )2, -OP(=O)(NR bb )2, -NR bb P(=O)(OR cc )2, -NR bb P(=O)(NR bb )2, -P(R cc )2, -P(R cc )3, -OP(R cc )2, -OP(R cc )3, -B(R aa )2, -B(OR cc )2, -BR aa (OR cc ), C1~C 50 Alkyl, C2~C 50 Alkenyl, C2~C 50 Alkinyl, C3~C 14 Carbocyclyl, 3-14 member heterocyclyl, C6-C 14 Examples include aryls and 5-14 member heteroaryls, where each alkyl, alkenyl, alkynyl, carbocykrill, heterocyclyl, aryl, and heteroaryl independently has 0, 1, 2, 3, 4, or 5 R dd Substituted with the base; Alternatively, the two geminal hydrogens on a carbon atom form the group =O, =S, =NN(R bb )2, =NNR bb C(=O)R aa ,=NNR bb C(=O)OR aa ,=NNR bb S(=O)2R aa ,=NR bb , or =NOR cc Replaced by, R aa Each example is independent of C1~C 50 Alkyl, C2~C50 Alkenyl, C2~C 50 Alkinyl, C3~C 10 Carbocyclyl, 3-14 member heterocyclyl, C6-C 14 Selected from aryls and 5-14 member heteroaryls, or two R aa The groups are linked together to form a 3-14 membered heterocyclyl or 5-14 membered heteroaryl ring, and each alkyl, alkenyl, alkynyl, carbocyryl, heterocyclyl, aryl, and heteroaryl independently has 0, 1, 2, 3, 4, or 5 R groups. dd Substituted with the base, R bb Each example independently represents hydrogen, -OH, and -OR. aa , -N(R cc )2, -CN, -C(=O)R aa -C(=O)N(R cc )2, -CO2R aa , -SO2R aa -C(=NR cc )OR aa -C(=NR cc )N(R cc )2, -SO2N(R cc )2, -SO2R cc , -SO2OR cc -SOR aa -C(=S)N(R cc )2, -C(=O)SR cc -C(=S)SR cc -P(=O)2R aa -P(=O)(R aa )2, -P(=O)2N(R cc )2, -P(=O)(NR cc )2, C1~C 50 Alkyl, C2~C 50 Alkenyl, C2~C 50 Alkinyl, C3~C 10 Carbocyclyl, 3-14 member heterocyclyl, C6-C 14 Selected from aryls and 5-14 member heteroaryls, or two R bbThe groups, together with the heteroatoms to which they are bonded, form a 3- to 14-membered heterocyclyl or 5- to 14-membered heteroaryl ring, and each alkyl, alkenyl, alkynyl, carbocykryl, heterocyclyl, aryl, and heteroaryl independently has 0, 1, 2, 3, 4, or 5 R groups. dd Substituted with the base; R cc Each example is independently of hydrogen, C1~C 50 Alkyl, C2~C 50 Alkenyl, C2~C 50 Alkinyl, C3~C 10 Carbocyclyl, 3-14 member heterocyclyl, C6-C 14 Selected from aryls and 5-14 member heteroaryls, or two R cc The groups, together with the heteroatoms to which they are bonded, form a 3- to 14-membered heterocyclyl or 5- to 14-membered heteroaryl ring, and each alkyl, alkenyl, alkynyl, carbocykryl, heterocyclyl, aryl, and heteroaryl independently has 0, 1, 2, 3, 4, or 5 R groups. dd Substituted with the base, R dd Each example is independently halogen, -CN, -NO2, -N3, -SO2H, -SO3H, -OH, -OR ee , -ON(R ff )2, -N(R ff )2, -N(R ff )3+X - , -N(OR ee )R ff -SH, -SR ee -SSR ee -C(=O)R ee -CO2H, -CO2R ee -OC(=O)R ee , -OCO2R ee -C(=O)N(R ff )2, -OC(=O)N(R ff )2, -NR ff C(=O)R ee , -NR ff CO2R ee , -NR ff C(=O)N(R ff )2, -C(=NRff )OR ee -OC(=NR ff )R ee -OC(=NR ff )OR ee -C(=NR ff )N(R ff )2, -OC(=NR ff )N(R ff )2, -NR ff C(=NR ff )N(R ff )2, -NR ff SO2R ee , -SO2N(R ff )2, -SO2R ee , -SO2OR ee , -OSO2R ee -S(=O)R ee , -Si(R ee )3, -OSi(R ee )3, -C(=S)N(R ff )2, -C(=O)SR ee -C(=S)SR ee -SC(=S)SR ee -P(=O)2R ee -P(=O)(R ee )2, -OP(=O)(R ee )2, -OP(=O)(OR ee )2, C1~C 50 Alkyl, C2~C 50 Alkenyl, C2~C 50 Alkinyl, C3~C 10 Carbocyclyl, 3-10 member heterocyclyl, C6-C 10 Selected from aryls and 5-10 membered heteroaryls, each alkyl, alkenyl, alkynyl, carbocykrill, heterocyclyl, aryl, and heteroaryl independently has 0, 1, 2, 3, 4, or 5 R gg Substituted by or two geminal R dd Substituents can be joined together to form =O or =S; R ee Each example is independent of C1~C 50 Alkyl, C2~C 50 Alkenyl, C2~C 50 Alkinyl, C3~C10 Carbocyclyl, C6~C 10 Selected from aryls, 3-10 membered heterocyclyls, and 3-10 membered heteroaryls, each alkyl, alkenyl, alkynyl, carbocykrill, heterocyclyl, aryl, and heteroaryl independently has 0, 1, 2, 3, 4, or 5 R gg Substituted with the base, R ff Each example is independently of hydrogen, C1~C 50 Alkyl, C2~C 50 Alkenyl, C2~C 50 Alkinyl, C3~C 10 Carbocyclyl, 3-10 member heterocyclyl, C6-C 10 Selected from aryls and 5-10 member heteroaryls, or two R ff The groups, together with the heteroatoms to which they are bonded, form a 3- to 14-membered heterocyclyl or 5- to 14-membered heteroaryl ring, and each alkyl, alkenyl, alkynyl, carbocykryl, heterocyclyl, aryl, and heteroaryl independently has 0, 1, 2, 3, 4, or 5 R groups. gg Substituted with the base, and R gg Each example is independently halogen, -CN, -NO2, -N3, -SO2H, -SO3H, -OH, -OC1~C 50 Alkyl, -ON(C1~C 50 Alkyl)2,-N(C1~C 50 Alkyl)2,-N(C1~C 50 Alkyl)3+X - -NH(C1~C 50 Alkyl)2+X - -NH2(C1~C 50 Alkyl) + X - -NH3+X - , -N(OC1~C 50 Alkyl)(C1~C 50 Alkyl), -N(OH)(C1~C 50 Alkyl), -NH(OH), -SH, -SC1~C 50 Alkyl, -SS(C1~C 50 Alkyl), -C(=O)(C1~C 50Alkyl), -CO2H, -CO2(C1~C 50 Alkyl), -OC(=O)(C1~C 50 Alkyl), -OCO2(C1~C 50 Alkyl), -C(=O)NH2, -C(=O)N(C1~C 50 Alkyl)2,-OC(=O)NH(C1~C 50 Alkyl), -NHC(=O)(C1~C 50 Alkyl), -N(C1~C 50 Alkyl)C(=O)(C1~C 50 Alkyl), -NHCO2(C1~C 50 Alkyl), -NHC(=O)N(C1~C 50 Alkyl)2,-NHC(=O)NH(C1~C 50 Alkyl), -NHC(=O)NH2, -C(=NH)O(C1~C 50 Alkyl), -OC (=NH) (C1~C 50 Alkyl), -OC(=NH)OC1~C 50 Alkyl, -C(=NH)N(C1~C 50 Alkyl)2, -C(=NH)NH(C1~C 50 Alkyl), -C(=NH)NH2, -OC(=NH)N(C1~C 50 Alkyl)2,-OC(NH)NH(C1~C 50 Alkyl), -OC(NH)NH2, -NHC(NH)N(C1~C 50 Alkyl)2, -NHC(=NH)NH2, -NHSO2(C1~C 50 Alkyl), -SO2N(C1~C 50 Alkyl)2,-SO2NH(C1~C 50 Alkyl), -SO2NH2, -SO2(C1~C 50 Alkyl), -SO2O(C1~C 50 Alkyl), -OSO2 (C1~C6 alkyl), -SO (C1~C6 alkyl), -Si (C1~C 50 Alkyl)3, -OSi(C1~C6 alkyl)3, -C(=S)N(C1~C 50 Alkyl)2, C(=S)NH(C1~C 50Alkyl), C(=S)NH2, -C(=O)S(C1~C6 alkyl), -C(=S)S(C1~C6 alkyl), -SC(=S)S(C1~C6 alkyl), -P(=O)2(C1~C 50 Alkyl), -P(=O)(C1~C 50 Alkyl)2, -OP(=O)(C1~C 50 Alkyl)2, -OP(=O)(OC1~C 50 Alkyl)2, C1~C 50 Alkyl, C2~C 50 Alkenyl, C2~C 50 Alkinyl, C3~C 10 Carbocyclyl, C6~C 10 It is an aryl, a 3-10 member heterocyclyl, a 5-10 member heteroaryl; or two geminal R gg Substituents may join together to form =O or =S; X - It is a counterion.

[0198] As used herein, the terms "halo" or "halogen" refer to fluorine (fluoro, -F), chlorine (chloro, ~Cl), bromine (bromo, -Br), or iodine (iod, -I).

[0199] As used herein, “counterion” is a negatively charged group that binds to a positively charged quaternary amine to maintain electronic neutrality. Exemplary counterions include halide ions (e.g., F - Cl - , Br - , I - ), NO3 - ClO4 - , OH - H2PO4 - HSO4 -Examples include sulfonate ions (e.g., methanesulfonic acid, trifluoromethanesulfonic acid, p-toluenesulfonic acid, benzenesulfonic acid, 10-camphorsulfonic acid, naphthalene-2-sulfonic acid, naphthalene-1-sulfonic acid-5-sulfonic acid, ethane-1-sulfonic acid-2-sulfonic acid, etc.) and carboxylate ions (e.g., acetic acid, ethaneic acid, propanoic acid, benzoic acid, glyceric acid, lactic acid, tartaric acid, glycolic acid, etc.).

[0200] Nitrogen atoms may be substituted or unsubstituted as their valence allows, and include primary, secondary, tertiary, and quaternary nitrogen atoms. Exemplary nitrogen atom substituents include, but are not limited to, hydrogen, -OH, and -OR. aa , -N(R cc )2, -CN, -C(=O)R aa -C(=O)N(R cc )2, -CO2R aa , -SO2R aa -C(=NR bb )R aa -C(=NR cc )OR aa -C(=NR cc )N(R cc )2, -SO2N(R cc )2, -SO2R cc , -SO2OR cc -SOR aa -C(=S)N(R cc )2, -C(=O)SR cc -C(=S)SR cc -P(=O)2R aa -P(=O)(R aa )2, -P(=O)2N(R cc )2, -P(=O)(NR cc )2, C1~C 50 Alkyl, C2~C 50 Alkenyl, C2~C 50 Alkinyl, C3~C 10 Carbocyclyl, 3-14 member heterocyclyl, C6-C 14 Examples include aryls and 5-14 member heteroaryls, or two R ccThe groups, together with the N atom to which they are bonded, form a 3- to 14-membered heterocyclyl or 5- to 14-membered heteroaryl ring, and each alkyl, alkenyl, alkynyl, carbocykryl, heterocyclyl, aryl, and heteroaryl independently has 0, 1, 2, 3, 4, or 5 R groups. dd Substituted with R aa , R bb , R cc and R dd This is as defined above.

[0201] In certain embodiments, the substituent present on the nitrogen atom is a nitrogen protecting group (also known as an amino protecting group). Nitrogen protecting groups are well known in the art and include those described in detail in Protecting Groups in Organic Synthesis, TW Greene and PGMWuts, 3rd edition, John Wiley & Sons, 1999, which is incorporated herein by reference.

[0202] For example, an amide group (for example, -C(=O)R aa Examples of nitrogen protecting groups include, but are not limited to, formamide, acetamide, chloroacetamide, trichloroacetamide, trifluoroacetamide, phenylacetamide, 3-phenylpropanamide, picolinamide, 3-pyridylcarboxamide, N-benzoylphenylalanyl derivatives, benzamide, p-phenylbenzamide, o-nitrophenylacetamide, o-nitrophenoxyacetamide, acetacetamide (N'-dithiobenzyloxyacylamino)acetamide, 3-(p-hydroxyphenyl)propanamide, 3-(o-nitrophenyl)propanamide, 2-methyl-2-(o-nitrophenoxy)propanamide, 2-methyl-2-(o-phenylazofenoxy)propanamide, 4-chlorobutanamide, 3-methyl-3-nitrobutanamide, o-nitrocinnamide, N-acetylmethionine derivatives, o-nitrobenzamide, and o-(benzoyloxymethyl)benzamide.

[0203] Carbamate group (e.g., -C(=O)OR) aaThe nitrogen protecting groups include, but are not limited to, methyl carbamate, ethyl carbamante, 9-fluorenyl methyl carbamate (Fmoc), 9-(2-sulfo)fluorenyl methyl carbamate, 9-(2,7-dibromo)fluorenyl methyl carbamate, 2,7-di-t-butyl-[9-(10,10-dioxo-10,10,10,10-tetrahydrothioxantyl)]methyl carbamate (DBD-Tmoc), 4-methoxyphenacyl carbamate (Phenoc), and 2,2,2-trichloroethyl carbamate. Rubamate (Troc), 2-trimethylsilylethylcarbamate (Teoc), 2-phenylethylcarbamate (hZ), 1-(1-adamanty1)-1-methylethylcarbamate (Adpoc), 1,1-dimethyl-2-haloethylcarbamate, 1,1-dimethyl-2,2-dibromoethylcarbamate (DB-t-BOC), 1,1-dimethyl-2,2,2-trichloroethylcarbamate (TCBOC), 1-methyl 1-(4-biphenylyl)ethyl carbamate (Bpoc), 1-(3,5-di-t-butylphenyl)-1-methylethyl carbamate (t-Bumeoc), 2-(2'-and 4'-pyridyl)ethyl carbamate (Pyoc), 2-(N,N-dicyclohexylcarboxamide)ethyl carbamate, t-butyl carbamate (BOC), 1-adamantyl carbamate (Adoc), vinyl carbamate (Voc), allyl carbamate (A lloc), 1-isopropylallyl carbamate (Ipaoc), cinnamyl carbamate (Coc), 4-nitrocinnamyl carbamate (Noc), 8-quinolyl carbamate, N-hydroxypiperidinyl carbamate, alkyl dithiocarbamate, benzyl carbamate (Cbz), p-methoxybenzyl carbamate (Moz), p-nitrobenzyl carbamate, p-bromobenzyl carbamate, p-chlorobenzyl carbamate, 2,4-Dichlorobenzylcarbamate, 4-Methylsulfinylbenzylcarbamate (Msz), 9-Anthrylmethylcarbamate, Diphenylmethylcarbamate, 2-Methylthioethylcarbamate, 2-Methylsulfonylethylcarbamate, 2-(p-Toluenesulfonyl)ethylcarbamate, [2-(1,3-Dithianyl)]methylcarbamate (Dmoc), 4-Methylthiophenylcarbamate (Mtpc), 2,4-Dimethylthiophenylcarbamate (Bmpc), 2-Phosphosphoethylcarbamate Peoc, 2-triphenylphosphonioisopropylcarbamate (Ppoc), 1,1-dimethyl-2-cyanoethylcarbamate, m-chloro-p-acyloxybenzylcarbamate, p-(dihydroxyboryl)benzylcarbamate, 5-benzisoxazolylmethylcarbamate, 2-(trifluoromethyl)-6-chromonylmethylcarbamate (Tcroc), m-nitrophenylcarbamate, 3,5-dimethoxybenzylcarbamate, o-nitrobenzylcarbamate, 3,4-di Methoxy-6-nitrobenzylcarbamate, phenyl(o-nitrophenyl)methylcarbamate, t-amylcarbamate, S-benzylthiocarbamate, p-cyanobenzylcarbamate, cyclobutylcarbamate, cyclohexylcarbamate, cyclopentylcarbamate, cyclopropylmethylcarbamate, p-decyloxybenzylcarbamate, 2,2-dimethoxyacylvinylcarbamate, o-(N,N-dimethylcarboxamide)benzylcarbamate, 1,1-dimethyl-3-(N,N -Dimethylcarboxamide)propylcarbamate, 1,1-dimethylpropynylcarbamate, di(2-pyridyl)methylcarbamate, 2-furanylmethylcarbamate, 2-iodoethylcarbamate, isobornylcarbamate, isobutylcarbamate, isonicotinylcarbamate, p-(p'-methoxyphenylazo)benzylcarbamate, 1-methylcyclobutylcarbamate, 1-methylcyclohexylcarbamate, 1-methyl-l-cyclopropylmethylcarbamate, 1-methyl-1(3,Examples include 5-dimethoxyphenyl)ethyl carbamate, 1-methyl-1-(p-phenylazophenyl)ethyl carbamate, 1-methyl-l-phenylethyl carbamate, 1-methyl-1-(4-pyridyl)ethyl carbamate, phenyl carbamate, p-(phenylazo)benzyl carbamate, 2,4,6-tri-t-butylphenyl carbamate, 4-(trimethylammonium)benzyl carbamate, and 2,4,6-trimethylbenzyl carbamate.

[0204] Sulfonamide group (e.g., -S(=O)2R) aa The nitrogen protecting groups include, but are not limited to, p-toluenesulfonamide (Ts), benzenesulfonamide, 2,3,6-trimethyl-4-methoxybenzenesulfonamide (Mtr), 2,4,6-trimethoxybenzenesulfonamide (Mtb), 2,6-dimethyl-4-methoxybenzenesulfonamide (Pme), 2,3,5,6-tetramethyl-4-methoxybenzenesulfonamide (Mte), 4-methoxybenzenesulfonamide (Mbs), and 2,4,6-trimethylbenzenesulfonamide. Examples include phonamide (Mts), 2,6-dimethoxy-4-methylbenzenesulfonamide (iMds), 2,2,5,7,8-pentamethylchroman-6-sulfonamide (Pmc), methanesulfonamide (Ms), β-trimethylsilylethanesulfonamide (SES), 9-anthracenesulfonamide, 4-(4',8'-dimethoxynaphthylmethyl)benzenesulfonamide (DNMBS), benzylsulfonamide, trifluoromethylsulfonamide, and phenacylsulfonamide.

[0205] Other nitrogen protecting groups include, but are not limited to, phenothiazinyl-(10)-acyl derivatives, N'-p-toluenesulfonylaminoacyl derivatives, N'-phenylaminothioacyl derivatives, N-benzoylphenylalanyl derivatives, N-acetylmethionine derivatives, 4,5-diphenyl-3-oxazolin-2-one, N-phthalimide, N-dithiasuccinimide (Dts), N-2,3-diphenylmaleimide, N-2,5-dimethylpyrrole, and N-1,1,4,4-tetramethyldisili Luazacyclopentane adduct (STABASE), 5-substituted 1,3-dimethyl-1,3,5-triazacyclohexane-2-one, 5-substituted 1,3-dibenzyl-1,3,5-triazacyclohexane-2-one, 1-substituted 3,5-dinitro-4-pyridone, N-methylamine, N-allylamine, N-[2-(trimethylsilyl)ethoxy]methylamine (SEM), N-3-acetoxypropylamine, N-(1-isopropyl-4-nitro-2-oxo-3-pyroline (pyroolin) )-3-yl)amine, quaternary ammonium salt, N-benzylamine, N-di(4-methoxyphenyl)methylamine, N-5-dibenzosberylamine, N-triphenylmethylamine (Tr), N-[(4-methoxyphenyl)diphenylmethyl]amine (MMTr), N-9-phenylfluorenylamine (PhF), N-2,7-dichloro-9-fluorenylmethyleneamine, N-ferrocenylmethylamino (Fcm), N-2-picolylamino N'-oxide, N-1,1-dimethylamine N-Tylthiomethyleneamine, N-Benzylideneamine, Np-Methoxybenzylideneamine, N-Diphenylmethyleneamine, N-[(2-Pyridyl)Mesityl]methyleneamine, N-(N',N'-Dimethylaminomethylene)amine, N,N'-Isopropylidenediamine, Np-Nitrobenzylideneamine, N-Salicylideneamine, N-5-Chlorosalicylideneamine, N-(5-Chloro-2-Hydroxyphenyl)phenylmethyleneamine, N-Cyclohexylideneamine, N-(5,Examples include 5-dimethyl-3-oxo-l-cyclohexenyl)amine, N-borane derivatives, N-diphenylboric acid derivatives, N-[phenyl(pentaacylchromium- or tungsten)acyl]amine, N-copper chelate, N-zinc chelate, N-nitroamine, N-nitrosamine, amine N-oxide, diphenylphosphinamide (Dpp), dimethylthiophosphinamide (Mpt), diphenylthiophosphinamide (Ppt), dialkylphosphoramide, dibenzylphosphoramide, diphenylphosphoramide, benzenesulfenamide, o-nitrobenzenesulfenamide (Nps), 2,4-dinitrobenzenesulfenamide, pentachlorobenzenesulfenamide, 2-nitro-4-methoxybenzenesulfenamide, triphenylmethylsulfenamide, and 3-nitropyridinesulfenamide (Npys).

[0206] In certain embodiments, the substituent present on the oxygen atom is an oxygen protecting group (also known as a hydroxyl protecting group). Oxygen protecting groups are well known in the art and include those described in detail in Protecting Groups in Organic Synthesis, TW Greene and PGMWuts, 3rd edition, John Wiley & Sons, 1999, which is incorporated herein by reference.

[0207] Examples of oxygen protecting groups include, but are not limited to, methyl, methoxymethyl (MOM), methylthiomethyl (MTM), t-butylthiomethyl, (phenyldimethylsilyl)methoxymethyl (SMOM), benzyloxymethyl (BOM), p-methoxybenzyloxymethyl (PMBM), (4-methoxyphenoxy)methyl (p-AOM), guaiacolmethyl (GUM), t-butoxymethyl, 4-pentenyloxymethyl (POM), siloxymethyl, 2-methoxyethoxymethyl ( MEM), 2,2,2-trichloroethoxymethyl, bis(2-chloroethoxy)methyl, 2-(trimethylsilyl)ethoxymethyl (SEMOR), tetrahydropyranyl (THP), 3-bromotetrahydropyranyl, tetrahydrothiopyranyl, 1-methoxycyclohexyl, 4-methoxytetrahydropyranyl (MTHP), 4-methoxytetrahydrothiopyranyl, 4-methoxytetrahydrothiopyranyl S,S-dioxide, 1-[(2-chloro-4-methyl)phenyl]-4-meth Xypiperidine-4-yl(y1)(CTMP), 1,4-dioxan-2-yl, tetrahydrofuranyl, tetrahydrothiofuranyl, 2,3,3a,4,5,6,7,7a-octahydro-7,8,8-trimethyl-4,7-methanobenzofuran-2-yl, 1-ethoxyethyl, 1-(2-chloroethoxy)ethyl, 1-methyl-l-methoxyethyl, 1-methyl-1-benzyloxyethyl, 1-methyl-1-benzyloxy-2-fluoroethyl, 2,2,2-trichloroethyl, 2 -Trimethylsilylethyl, 2-(phenylselenyl)ethyl, t-butyl, allyl, p-chlorophenyl, p-methoxyphenyl, 2,4-dinitrophenyl, benzyl (Bn), p-methoxybenzyl, 3,4-dimethoxybenzyl, o-nitrobenzyl, p-nitrobenzyl, p-halobenzyl, 2,6-dichlorobenzyl, p-cyanobenzyl, p-phenylbenzyl, 2-picolyl, 4-picolyl, 3-methyl-2-picolyl (picoly1)N-oxide, diphenylmethyl, p,p'-Dinitrobenzhydryl, 5-Dibenzosberyl, Triphenylmethyl, α-Naphthyldiphenylmethyl, p-Methoxyphenyldiphenylmethyl, Di(p-Methoxyphenyl)phenylmethyl, Tri(p-Methoxyphenyl)methyl, 4-(4'-Bromophenacyloxyphenyl)diphenylmethyl, 4,4',4”-Tris(4,5-Dichlorophthalimidophenyl)methyl, 4,4',4”-Tris(Lebrinoyloxyphenyl)methyl, 4,4',4”-Tris(Benzoyloxyphenyl)methyl, 3-(Imi Dazole-1-yl)bis(4',4"-dimethoxyphenyl)methyl, 1,1-bis(4-methoxyphenyl)-1'-pyrenylmethyl, 9-anthryl, 9-(9-phenyl)xanthenyl, 9-(9-phenyl-10-oxo)anthryl, 1,3-benzodisulfuran-2-yl, benzisothiazolyl S,S-dioxide, trimethylsilyl (TMS), triethylsilyl (TES), triisopropylsilyl (TIPS), dimethylisopropylsilyl (IPDMS), diethylisopropylsilyl (DEIPS), di Methyltexylsilyl, t-butyldimethylsilyl (TBDMS), t-butyldiphenylsilyl (butyldiphenylsily1) (TBDPS), tribenzylsilyl, tri-p-xylylsilyl, triphenylsilyl, diphenylmethylsilyl (DPMS), t-butylmethoxyphenylsilyl (TBMPS), formate, benzoylformate, acetate, chloroacetate, dichloroacetate, trichloroacetate, trifluoroacetate, methoxyacetate, triphenylmethoxyacetate, f Phenoxyacetate, p-chlorophenoxyacetate, 3-phenylpropionate, 4-oxopentanoate (rebrinate), 4,4-(ethylenedithio)pentanoate (rebrinoyldithioacetal), pivaloate, adamantoate, crotonate, 4-methoxycrotonate, benzoate, p-phenylbenzoate, 2,4,6-trimethylbenzoate (mesitoate), alkylmethyl carbonate, 9-fluorenylmethyl carbonate (Fmoc), alkylethyl carbonate, alkyl 2,22-Trichloroethyl carbonate (Troc), 2-(trimethylsilyl)ethyl carbonate (TMSEC), 2-(phenylsulfonyl)ethyl carbonate (Psec), 2-(triphenylphosphonio)ethyl carbonate (Peoc), alkyl isobutyl carbonate, alkyl vinyl carbonate, alkyl allyl carbonate, alkyl p-nitrophenyl carbonate, alkyl benzyl carbonate, alkyl p-methoxybenzyl carbonate, alkyl 3,4-dimethoxybenzyl carbonate, alkyl o-nitrobenzyl carbonate, alkyl p-nitrobenzyl carbonate, alkyl S-benzylthiocarbonate, 4-ethoxy-1-naphthyl carbonate, methyl dithiocarbonate, 2-iodobenzoate, 4-azidobutyrate, 4-nitro-4-methylpentanoate, o-(dibromomethyl)benzoate, 2-formylbe Phenoxyethanolate, 2-(methylthiomethoxy)ethyl, 4-(methylthiomethoxy)butyrate, 2-(methylthiomethoxymethyl)benzoate, 2,6-dichloro-4-methylphenoxyacetate, 2,6-dichloro-4-(1,1,3,3-tetramethylbutyl)phenoxyacetate, 2,4-bis(1,1-dimethylpropyl)phenoxyacetate, chlorodiphenylacetate, isobutyrate, monosuccinoate Examples include (E)-2-methyl-2-butenoate, o-(methoxyacyl)benzoate, α-naphthoate, nitrate, alkyl N,N,N',N'-tetramethylphosphodiamide, alkyl N-phenylcarbamate, borate, dimethylphosphinthiole, alkyl 2,4-dinitrophenyl sulfenate, sulfate, methanesulfonate (mesylate), benzylsulfonate, and tosylate (Ts).

[0208] In certain embodiments, substituents on the sulfur atom are sulfur protecting groups (also known as thiol protecting groups). Sulfur protecting groups are well known in the art and include those described in detail in Protecting Groups in Organic Synthesis, TW Greene and PGMWuts, 3rd edition, John Wiley & Sons, 1999, which is incorporated herein by reference.

[0209] Examples of sulfur protecting groups include alkyl, benzyl, p-methoxybenzyl, 2,4,6-trimethylbenzyl, 2,4,6-trimethoxybenzyl, o-hydroxybenzyl, p-hydroxybenzyl, o-acetoxybenzyl, p-acetoxybenzyl, p-nitrobenzyl, 4-picolyl, 2-quinolinylmethyl, 2-picolyl N-oxide, 9-anthrylmethyl, 9-fluorenylmethyl, xanthenyl, ferrocenylmethyl, diphenylmethyl, bis(4-methoxyphenyl)methyl, 5-dibenzosberyl, triphenylmethyl, diphenyl-4-pyridylmethyl, phenyl, 2,4-dinitrophenyl, t-butyl, 1-adamantyl, methoxymethyl (MOM), isobutoxymethyl, benzyloxymethyl, 2-tetrahydropyranyl, benzylthiomethyl, phenylthiomethyl, thiazolidino, acetamidomethyl, trimethylacetamidomethyl, benzamidomethyl, allyloxycarbonylaminomethyl, phenyl Nylacetamidomethyl, phthalimidomethyl, acetylmethyl, carboxymethyl, cyanomethyl(2-nitro-1-phenyl)ethyl, 2-(2,4-dinitrophenyl)ethyl, 2-cyanoethyl, 2-(trimethylsilyl)ethyl, 2,2-bis(carboethoxy)ethyl(1-m-nitrophenyl-2-benzoyl)ethyl, 2-phenylsulfonylethyl, 2-(4-methylphenylsulfonyl)-2-methylpropane-2-yl, acetyl, benzoyl, t Examples include, but are not limited to, difluoroacetyl, N-[[(p-biphenylyl)isopropoxy]carbonyl]-N-methyl]-γ-aminothiobutyrate, 2,2,2-trichloroethoxycarbonyl, t-butoxycarbonyl, benzyloxycarbonyl, p-methoxybenzyloxycarbonyl, N-ethyl, N-methoxymethyl, sulfonates, sulfenylthiocarbonates, 3-nitro-2-pyridinesulfenylsulfide, and oxathiolone.

[0210] Compound of the present invention Liposome-based media are considered attractive carriers for therapeutic drugs and remain the subject of ongoing development efforts. While liposome-based vehicles containing specific lipid components have shown promising results in terms of encapsulation, stability, and site localization, there is still a great need for improvements to liposome-based delivery systems. For example, significant shortcomings of liposome delivery systems relate to the construction of liposomes with sufficient cell culture or in vivo stability to reach desired target cells and / or intracellular compartments, and the ability of such liposome delivery systems to efficiently release encapsulated material into such target cells.

[0211] Therefore, the efficient delivery of liposome-encapsulated nucleic acids remains an active area of ​​research. During liposome loading, cationic lipid components play a crucial role in promoting effective nucleic acid encapsulation. In addition, cationic lipids can play a vital role in the efficient release of nucleic acid cargo from liposomes into the cytoplasm of target cells. Various cationic lipids suitable for in vivo use have been discovered. However, there is still a need to identify lipids that can be synthesized efficiently and inexpensively without forming potentially toxic byproducts.

[0212] More specifically, there is still a need for improved lipid compounds that demonstrate enhanced pharmacokinetic properties and possess the ability to deliver macromolecules such as nucleic acids to a wide range of cell types and tissues with improved efficiency. Importantly, there is also still a need for novel lipid compounds that are characterized in particular by reduced toxicity and the ability to efficiently deliver encapsulated nucleic acids and polynucleotides to target cells, tissues, and organs.

[0213] A novel class of tricine and citrate-based cationic lipid compounds for improving the in vivo delivery of therapeutic agents such as nucleic acids is described herein. More specifically, the tricine and citrate-based cationic lipids described herein can be optionally used as cationic lipids in combination with other lipids to formulate lipid-based nanoparticles (e.g., liposomes) for encapsulating therapeutic agents, such as nucleic acids (e.g., DNA, siRNA, mRNA, microRNA), for therapeutic use.

[0214] In embodiments, the compounds of the present invention described herein can provide one or more desired features or properties. That is, in certain embodiments, the compounds of the present invention described herein can be characterized by having one or more properties that give such compounds an advantage compared to other similarly classified lipids. For example, the compounds disclosed herein can enable the control and modification of the properties of liposome compositions (e.g., lipid nanoparticles) in which they are components. In particular, the compounds disclosed herein can be characterized by enhanced transfection efficiency and their ability to induce specific biological outcomes. Such outcomes may include, for example, enhanced cellular uptake, endosomal / lysosome disruption ability and / or enhanced intracellular release of encapsulated materials (e.g., polynucleotides). In addition, the compounds disclosed herein have advantageous pharmacokinetic properties, biodistribution, and efficiency (e.g., due to different dissociation rates of the polymer groups used).

[0215] Examples of compounds are those described herein. The variables described for a formula may be any other formula or embodiment described herein, and any combination thereof, of any acceptable structure or value.

[0216] Formula (A2) In one embodiment, the present invention relates to a structure of formula (A2): [ka] The formula is characterized by a cationic lipid having or a pharmaceutically acceptable salt thereof, wherein X is independently either O or NH; R 1 , R 2 , and R 3 Each of these independently corresponds to C4~C 30 Alkyl, C4~C 30 Alkenyl, C4~C 30 Alkinyl, or C4~C 30 It is heteroalkyl; A is [ka] It is a substructure selected from, B is independently an ionizable nitrogen-containing group or a permanently charged nitrogen group; m is an integer between 2 and 10; n is an integer between 2 and 10; L 1 is a carbonyl, ester, or amide; L 2 C2~C 10 Alkylene or C2~C 10 It is an alkenylene; Ar is a phenylene compound optionally containing 1 to 4 substituents independently selected from halogens, OCH3, and CH3; R 4 C1~C 10 It is alkylene; Here, A is R 1 , R 2 , and R 3 Each of them independently contains a disulfide bond, or [ka] C4~C has the structure 30 If it is a heteroalkyl, it is the only substructure (a4), L 3 is OC(O), CO2, or (O)CO; o is an integer between 2 and 5; R 5 C4~C 24 Alkyl; or L3 is OC(O), CO2, or (O)CO; o is an integer between 6 and 12; R 5 These are C1-C6 alkyl groups.

[0217] In this embodiment, A is R 1 , R 2 , and R 3 Each of them independently contains a disulfide bond, or [ka] C4~C has the structure 30 If it is a heteroalkyl, then it is the only substructure (a4), where L 3 is OC(O), CO2, or (O)CO; o is an integer between 2 and 5; R 5 C4~C 24 Alkyl; or L 3 is OC(O), CO2, or (O)CO; o is an integer between 6 and 12; R 5 These are C1-C6 alkyl groups.

[0218] In this embodiment, X is independently O.

[0219] In this embodiment, X is independently NH.

[0220] In this embodiment, R 1 , R 2 and R 3 Each of these is independent of C4~C 30 It is alkyl. In this embodiment, R 1 , R 2 and R 3 Each of these is independently, non-substituted C4~C 30 It is alkyl. In this embodiment, R 1 , R 2 and R 3 Each of these independently substitutes C4~C 30 It is alkyl.

[0221] In this embodiment, R 1 , R 2 and R3 Each of these is independent of C4~C 30 It is an alkenyl. In this embodiment, R 1 , R 2 and R 3 Each of these is independently, non-substituted C4~C 30 It is an alkenyl. In this embodiment, R 1 , R 2 and R 3 Each of these independently substitutes C4~C 30 It is Alkenil.

[0222] In this embodiment, R 1 , R 2 and R 3 Each of these is independent of C4~C 30 It is alkynyl. In the embodiment, R 1 , R 2 and R 3 Each of these is independently, non-substituted C4~C 30 It is alkynyl. In the embodiment, R 1 , R 2 and R 3 Each of these independently substitutes C4~C 30 It is alkinyl.

[0223] In this embodiment, R 1 , R 2 and R 3 Each of these is independent of C4~C 30 Heteroalkyl (e.g., C6~C) 30 It is a heteroalkyl group. In this embodiment, R 1 , R 2 and R 3 Each of these is independently, non-substituted C4~C 30 Heteroalkyl (e.g., unsubstituted C6~C) 30 It is a heteroalkyl group. In this embodiment, R 1 , R 2 and R 3 Each of these independently substitutes C4~C 30 Heteroalkyl (e.g., substituted C6~C) 30 It is a heteroalkyl group.

[0224] In this embodiment, B is independently an ionizable nitrogen-containing group.

[0225] In this embodiment, B is independently a permanently charged nitrogen group.

[0226] In this embodiment, A is [ka] That is the case.

[0227] In this embodiment, A is [ka] That is the case.

[0228] In this embodiment, A is [ka] That is the case.

[0229] In this embodiment, A is [ka] That is the case.

[0230] In this embodiment, m is an integer between 2 and 10.

[0231] In this embodiment, n is an integer between 2 and 10.

[0232] In this embodiment, L 1 It is a carbonyl group.

[0233] In this embodiment, L 1 is an ester. In this embodiment, L 1 is -OC(O)-. In the embodiment, L 1 is -(O)CO-.

[0234] In this embodiment, L 1 is an amide. In this embodiment, L 1is -NHC(O)-. In the embodiment, L 1 It is -C(O)NH-.

[0235] In this embodiment, L 2 C2~C 10 It is an alkylene. In the embodiment, L 2 is unsubstituted C2~C 10 It is an alkylene. In the embodiment, L 2 This is the substitution C2~C 10 It is alkylene.

[0236] In this embodiment, L 2 C2~C 10 It is an alkenylene. In the embodiment, L 2 is unsubstituted C2~C 10 It is an alkenylene. In the embodiment, L 2 This is the substitution C2~C 10 It is alkenylene.

[0237] In the embodiment, Ar is phenylene containing 1 to 4 substituents independently selected from halogens, OCH3, and CH3. In the embodiment, Ar is unsubstituted phenylene. In the embodiment, Ar is phenylene containing 1 to 4 substituents independently selected from halogens, OCH3, and CH3. In the embodiment, Ar is phenylene containing 1 to 3 substituents independently selected from halogens, OCH3, and CH3. In the embodiment, Ar is phenylene containing 1 to 2 substituents independently selected from halogens, OCH3, and CH3. In the embodiment, Ar is phenylene containing 1 substituent selected from halogens, OCH3, and CH3.

[0238] In this embodiment, R 4 This is unsubstituted C1~C 10 It is an alkylene. Embodiment, R 4 This is the substitution C1~C 10 It is alkylene.

[0239] In the embodiment, X is O, and / or B is independently an ionizable nitrogen-containing group.

[0240] In this embodiment, R 1 , R 2 , and R 3 Each of these is independent of C4~C 30 Alkyl, C4~C 30 Alkenyl, or C4~C 30 It is alkynyl. In the embodiment, R 1 , R 2 and R 3 Each of them is non-substitutable. In this embodiment, R 1 , R 2 and R 3 Each of these is substituted (for example, including 1 to 3 substituents as described herein).

[0241] In this embodiment, R 1 , R 2 , and R 3 Each of them independently contains a disulfide bond, or [ka] C4~C has the structure 30 It is a heteroalkyl, where L 3 is OC(O), CO2, or (O)CO; o is an integer between 2 and 5; R 5 C4~C 24 Alkyl; or L 3 is OC(O), CO2, or (O)CO; o is an integer between 6 and 12; R 5 These are C1-C6 alkyl groups.

[0242] In this embodiment, B is independently, [ka] That is the case.

[0243] In this embodiment, B is independently, [ka] That is the case.

[0244] In this embodiment, B is independently, [ka] That is the case.

[0245] In this embodiment, B is independently, [ka] That is the case.

[0246] In this embodiment, B is independently, [ka] That is the case.

[0247] In this embodiment, B is independently, [ka] That is the case.

[0248] In this embodiment, B is independently, [ka] That is the case.

[0249] In this embodiment, B is independently, [ka] That is the case.

[0250] In this embodiment, B is independently, [ka] That is the case.

[0251] In this embodiment, B is independently, [ka] That is the case.

[0252] In this embodiment, B is independently, [ka] That is the case.

[0253] In this embodiment, B is independently, [ka] That is the case.

[0254] In this embodiment, B is independently, [ka] That is the case.

[0255] In this embodiment, B is independently, [ka] That is the case.

[0256] In this embodiment, B is independently, [ka] That is the case.

[0257] In this embodiment, B is independently, [ka] That is the case.

[0258] In this embodiment, B is independently, [ka] That is the case.

[0259] In this embodiment, B is independently, [ka] That is the case.

[0260] In this embodiment, B is independently, [ka] That is the case.

[0261] In this embodiment, B is independently, [ka] That is the case.

[0262] In this embodiment, B is independently, [ka] That is the case.

[0263] In this embodiment, B is independently, [ka] That is the case.

[0264] In this embodiment, B is independently, [ka] That is the case.

[0265] In this embodiment, B is independently, [ka] That is the case.

[0266] Formula (A1) In one embodiment, the present invention relates to a structure of formula (A1): [ka] The formula is characterized by a cationic lipid having or a pharmaceutically acceptable salt thereof, wherein X is independently either O or NH; R 1 , R 2 , and R 3 Each of these is independent of C4~C 30 Alkyl, C4~C 30 Alkenyl, C4~C 30 Alkinyl, or C4~C 30 It is heteroalkyl; A is [ka] It is a substructure selected from, B is independently an ionizable nitrogen-containing group or a permanently charged nitrogen group; m is an integer between 2 and 10; n is an integer between 2 and 10; L 1 is a carbonyl, ester, or amide; L 2 C2~C 10 Alkylene or C2~C 10 It is an alkenylene; Ar is a phenylene compound optionally containing 1 to 4 substituents independently selected from halogens, OCH3, and CH3; R 4 C1~C 10 It is alkylene; Here, A is R 1 , R 2 , and R 3 Each of them independently contains a disulfide bond, or [ka] C4~C has the structure 30 If it is a heteroalkyl, it is the only substructure (a4), L 3 is OC(O), CO2, or (O)CO; o is an integer between 2 and 5; R 5 C4~C 24 Alkyl; or L3 is OC(O), CO2, or (O)CO; o is an integer between 6 and 12; R 5 These are C1-C5 alkyl groups.

[0267] In this embodiment, A is R 1 , R 2 , and R 3 Each of them independently contains a disulfide bond, or [ka] C4~C has the structure 30 If it is a heteroalkyl, then it is the only substructure (a4), where L 3 is OC(O), CO2, or (O)CO; o is an integer between 2 and 5; R 5 C4~C 24 Alkyl; or L 3 is OC(O), CO2, or (O)CO; o is an integer between 6 and 12; R 5 These are C1-C5 alkyl groups.

[0268] In this embodiment, X is independently O.

[0269] In this embodiment, X is independently NH.

[0270] In this embodiment, R 1 , R 2 and R 3 Each of these is independent of C4~C 30 It is alkyl. In this embodiment, R 1 , R 2 and R 3 Each of these is independently, non-substituted C4~C 30 It is alkyl. In this embodiment, R 1 , R 2 and R 3 Each of these independently substitutes C4~C 30 It is alkyl.

[0271] In this embodiment, R 1 , R 2 and R3 Each of these is independent of C4~C 30 It is an alkenyl. In this embodiment, R 1 , R 2 and R 3 Each of these is independently, non-substituted C4~C 30 It is an alkenyl. In this embodiment, R 1 , R 2 and R 3 Each of these independently substitutes C4~C 30 It is Alkenil.

[0272] In this embodiment, R 1 , R 2 and R 3 Each of these is independent of C4~C 30 It is alkynyl. In the embodiment, R 1 , R 2 and R 3 Each of these is independently, non-substituted C4~C 30 It is alkynyl. In the embodiment, R 1 , R 2 and R 3 Each of these independently substitutes C4~C 30 It is alkinyl.

[0273] In this embodiment, R 1 , R 2 and R 3 Each of these is independent of C4~C 30 Heteroalkyl (e.g., C6~C) 30 It is a heteroalkyl group. In this embodiment, R 1 , R 2 and R 3 Each of these is independently, non-substituted C4~C 30 Heteroalkyl (e.g., unsubstituted C6~C) 30 It is a heteroalkyl group. In this embodiment, R 1 , R 2 and R 3 Each of these independently substitutes C4~C 30 Heteroalkyl (e.g., substituted C6~C) 30 It is a heteroalkyl group.

[0274] In this embodiment, B is independently an ionizable nitrogen-containing group.

[0275] In this embodiment, B is independently a permanently charged nitrogen group.

[0276] In this embodiment, A is [ka] That is the case.

[0277] In this embodiment, A is [ka] That is the case.

[0278] In this embodiment, A is [ka] That is the case.

[0279] In this embodiment, A is [ka] That is the case.

[0280] In this embodiment, m is an integer between 2 and 10.

[0281] In this embodiment, n is an integer between 2 and 10.

[0282] In this embodiment, L 1 It is a carbonyl group.

[0283] In this embodiment, L 1 is an ester. In this embodiment, L 1 is -OC(O)-. In the embodiment, L 1 is -(O)CO-.

[0284] In this embodiment, L 1 is an amide. In this embodiment, L 1is -NHC(O)-. In an embodiment, L 1 is -C(O)NH-.

[0285] In an embodiment, L 2 is C2 - C 10 alkylene. In an embodiment, L 2 is unsubstituted C2 - C 10 [[ID=!4]]alkylene. In an embodiment, L 2 is substituted C2 - C 10 alkylene.

[0286] In an embodiment, L 2 is C2 - C<000...900>alkenylene. In an embodiment, L 2 is unsubstituted C2 - C 10 alkenylene. In an embodiment, L<00...903>is substituted C2 - C 10 alkenylene.

[0287] In an embodiment, Ar is phenylene optionally containing 1 - 4 substituents independently selected from halogen, OCH3, and CH3. In an embodiment, Ar is unsubstituted phenylene. In an embodiment, Ar is phenylene containing​​​​​​​​​​​​​​​In the embodiment, X is O, and / or B is independently an ionizable nitrogen-containing group.

[0290] In this embodiment, R 1 , R 2 , and R 3 Each of these is independent of C4~C 30 Alkyl, C4~C 30 Alkenyl, or C4~C 30 It is alkynyl. In the embodiment, R 1 , R 2 and R 3 Each of them is non-substitutable. In this embodiment, R 1 , R 2 and R 3 Each of these is substituted (for example, including 1 to 3 substituents as described herein).

[0291] In this embodiment, R 1 , R 2 , and R 3 Each of them independently contains a disulfide bond, or [ka] C4~C has the structure 30 It is a heteroalkyl, where L 3 is OC(O), CO2, or (O)CO; o is an integer between 2 and 5; R 5 C4~C 24 Alkyl; or L 3 is OC(O), CO2, or (O)CO; o is an integer between 6 and 12; R 5 These are C1-C5 alkyl groups.

[0292] In this embodiment, B is independently, [ka] That is the case.

[0293] In this embodiment, B is independently, [ka] That is the case.

[0294] In this embodiment, B is independently, [ka] That is the case.

[0295] In this embodiment, B is independently, [ka] That is the case.

[0296] In this embodiment, B is independently, [ka] That is the case.

[0297] In this embodiment, B is independently, [ka] That is the case.

[0298] In this embodiment, B is independently, [ka] That is the case.

[0299] In this embodiment, B is independently, [ka] That is the case.

[0300] In this embodiment, B is independently, [ka] That is the case.

[0301] In this embodiment, B is independently, [ka] That is the case.

[0302] In this embodiment, B is independently, [ka] That is the case.

[0303] In this embodiment, B is independently, [ka] That is the case.

[0304] In this embodiment, B is independently, [ka] That is the case.

[0305] In this embodiment, B is independently, [ka] That is the case.

[0306] In this embodiment, B is independently, [ka] That is the case.

[0307] In this embodiment, B is independently, [ka] That is the case.

[0308] In this embodiment, B is independently, [ka] That is the case.

[0309] In this embodiment, B is independently, [ka] That is the case.

[0310] In this embodiment, B is independently, [ka] That is the case.

[0311] In this embodiment, B is independently, [ka] That is the case.

[0312] In this embodiment, B is independently, [ka] That is the case.

[0313] In this embodiment, B is independently, [ka] That is the case.

[0314] In this embodiment, B is independently, [ka] That is the case.

[0315] In this embodiment, B is independently, [ka] That is the case.

[0316] Formula (A) In one embodiment, the present invention relates to a structure of formula (A): [ka] The formula is characterized by a cationic lipid having or a pharmaceutically acceptable salt thereof, wherein X is independently either O or NH; R 1 , R 2 , and R 3 Each of these is independent of C4~C 30 Alkyl, C4~C 30 Alkenyl, C4~C 30 Alkinyl, or C4~C 30 It is heteroalkyl; A is [ka] It is a substructure selected from, B is independently an ionizable nitrogen-containing group or a permanently charged nitrogen group; m is an integer between 2 and 10; n is an integer between 2 and 10; L 1 is a carbonyl, ester, or amide; L 2 C2~C 10 Alkylene or C2~C 10 It is an alkenylene; Ar is a phenylene compound optionally containing 1 to 4 substituents independently selected from halogens, OCH3, and CH3; R 4 C2~C 10 It is alkylene.

[0317] In this embodiment, A is R 1 , R 2 , and R 3 Each of them independently contains a disulfide bond, or [ka] C4~C has the structure 30 If it is a heteroalkyl, it is the only substructure (a4), L 3 is OC(O), CO2, or (O)CO; o is an integer between 2 and 5; R 5 C4~C24 is alkyl; or L 3 is OC(O), CO2, or (O)CO; o is an integer from 6 to 12; R 5 is C1-C5 alkyl.

[0318] In an embodiment, X is independently O.

[0319] In an embodiment, X is independently NH. The

[0320] In an embodiment, R 1 , R 2 and R 3 each is independently C4-C 30 alkyl. In an embodiment, R 1 , R 2 and R 3 each is independently unsubstituted C4-C 30 alkyl. In an embodiment, R 1 , R 2 and R 3 each is independently substituted C4-C 30 alkyl.

[0321] The In an embodiment, R 1 , R 2 and R 3 each is independently C4-C 30 alkenyl. In an embodiment, R 1 , R 2 and R 3 each is independently unsubstituted C4-C 30 alkenyl. In an embodiment, R 1 , R 2 and R 3 each is independently substituted C4-C 30 alkenyl.

[0322] ]>In an embodiment, R 1 , R 2 and R 3 each is independently C4-C 30 alkynyl. In an embodiment, R[[ID=X]] 1 , R 2 and R3 Each of these is independently, non-substituted C4~C 30 It is alkynyl. In the embodiment, R 1 , R 2 and R 3 Each of these independently substitutes C4~C 30 It is alkinyl.

[0323] In this embodiment, R 1 , R 2 and R 3 Each of these is independent of C4~C 30 Heteroalkyl (e.g., C6~C) 30 It is a heteroalkyl group. In this embodiment, R 1 , R 2 and R 3 Each of these is independently, non-substituted C4~C 30 Heteroalkyl (e.g., unsubstituted C6~C) 30 It is a heteroalkyl group. In this embodiment, R 1 , R 2 and R 3 Each of these independently substitutes C4~C 30 Heteroalkyl (e.g., substituted C6~C) 30 It is a heteroalkyl group.

[0324] In this embodiment, B is independently an ionizable nitrogen-containing group.

[0325] In this embodiment, B is independently a permanently charged nitrogen group.

[0326] In this embodiment, A is [ka] That is the case.

[0327] In this embodiment, A is [ka] That is the case.

[0328] In this embodiment, A is [ka] That is the case.

[0329] In this embodiment, A is [ka] That is the case.

[0330] In this embodiment, m is an integer between 2 and 10.

[0331] In this embodiment, n is an integer between 2 and 10.

[0332] In this embodiment, L 1 It is a carbonyl group.

[0333] In this embodiment, L 1 is an ester. In this embodiment, L 1 is -OC(O)-. In the embodiment, L 1 is -(O)CO-.

[0334] In this embodiment, L 1 is an amide. In this embodiment, L 1 is -NHC(O)-. In the embodiment, L 1 It is -C(O)NH-.

[0335] In this embodiment, L 2 C2~C 10 It is an alkylene. In the embodiment, L 2 is unsubstituted C2~C 10 It is an alkylene. In the embodiment, L 2 This is the substitution C2~C 10 It is alkylene.

[0336] In this embodiment, L 2 C2~C 10 It is an alkenylene. In the embodiment, L 2 is unsubstituted C2~C 10 It is an alkenylene. In the embodiment, L 2 This is the substitution C2~C 10It is alkenylene.

[0337] In the embodiment, Ar is phenylene containing 1 to 4 substituents independently selected from halogens, OCH3, and CH3. In the embodiment, Ar is unsubstituted phenylene. In the embodiment, Ar is phenylene containing 1 to 4 substituents independently selected from halogens, OCH3, and CH3. In the embodiment, Ar is phenylene containing 1 to 3 substituents independently selected from halogens, OCH3, and CH3. In the embodiment, Ar is phenylene containing 1 to 2 substituents independently selected from halogens, OCH3, and CH3. In the embodiment, Ar is phenylene containing 1 substituent selected from halogens, OCH3, and CH3.

[0338] In this embodiment, R 4 is unsubstituted C2~C 10 It is an alkylene. In the embodiment, R 4 This is the substitution C2~C 10 It is alkylene.

[0339] In the embodiment, X is O, and / or B is independently an ionizable nitrogen-containing group.

[0340] In this embodiment, R 1 , R 2 , and R 3 Each of these is independent of C4~C 30 Alkyl, C4~C 30 Alkenyl, or C4~C 30 It is alkynyl. In the embodiment, R 1 , R 2 and R 3 Each of them is non-substitutable. In this embodiment, R 1 , R 2 and R 3 Each of these is substituted (for example, including 1 to 3 substituents as described herein).

[0341] In this embodiment, R1 , R 2 , and R 3 Each of them independently contains a disulfide bond, or [ka] C4~C has the structure 30 It is a heteroalkyl, where L 3 is OC(O), CO2, or (O)CO; o is an integer between 2 and 5; R 5 C4~C 24 Alkyl; or L 3 is OC(O), CO2, or (O)CO; o is an integer between 6 and 12; R 5 These are C1-C5 alkyl groups.

[0342] In this embodiment, B is independently, [ka] That is the case.

[0343] In this embodiment, B is independently, [ka] That is the case.

[0344] In this embodiment, B is independently, [ka] That is the case.

[0345] In this embodiment, B is independently, [ka] That is the case.

[0346] In this embodiment, B is independently, [ka] That is the case.

[0347] In this embodiment, B is independently, [ka] That is the case.

[0348] In this embodiment, B is independently, [ka] That is the case.

[0349] In this embodiment, B is independently, [ka] That is the case.

[0350] In this embodiment, B is independently, [ka] That is the case.

[0351] In this embodiment, B is independently, [ka] That is the case.

[0352] In this embodiment, B is independently, [ka] That is the case.

[0353] In this embodiment, B is independently, [ka] That is the case.

[0354] In this embodiment, B is independently, [ka] That is the case.

[0355] In this embodiment, B is independently, [ka] That is the case.

[0356] In this embodiment, B is independently, [ka] That is the case.

[0357] In this embodiment, B is independently, [ka] That is the case.

[0358] In this embodiment, B is independently, [ka] That is the case.

[0359] In this embodiment, B is independently, [ka] That is the case.

[0360] In this embodiment, B is independently, [ka] That is the case.

[0361] In this embodiment, B is independently, [ka] That is the case.

[0362] In this embodiment, B is independently, [ka] That is the case.

[0363] In this embodiment, B is independently, [ka] That is the case.

[0364] In this embodiment, B is independently, [ka] That is the case.

[0365] In this embodiment, B is independently, [ka] That is the case.

[0366] Equation (I) In the embodiment, the compound has the structure of formula (I): [ka] A salt having or a pharmaceutically acceptable salt thereof, wherein R 1 , R 2 , R 3 Each of B, m, and n independently follows any combination of embodiments described herein.

[0367] In this embodiment, R 1 , R 2 , and R 3 Each of these is independent of C6~C 30 Alkyl, C6~C 30 Alkenil, C6~C 30 Alkinyl, or C4~C 30 It is heteroalkyl; m is an integer between 2 and 10; n is an integer between 2 and 10; B is independently an ionizable nitrogen-containing group or a permanently charged nitrogen group.

[0368] In this embodiment, R1 , R 2 and R 3 Each of these is independent of C6~C 30 It is alkyl. In this embodiment, R 1 , R 2 and R 3 Each of these is independently, non-substituted C6~C 30 It is alkyl. In this embodiment, R 1 , R 2 and R 3 Each of these independently substitutes C6~C 30 It is alkyl.

[0369] In this embodiment, R 1 , R 2 and R 3 Each of these is independent of C6~C 30 It is an alkenyl. In this embodiment, R 1 , R 2 and R 3 Each of these is independently, non-substituted C6~C 30 It is an alkenyl. In this embodiment, R 1 , R 2 and R 3 Each of these independently substitutes C6~C 30 It is Alkenil.

[0370] In this embodiment, R 1 , R 2 and R 3 Each of these is independent of C6~C 30 It is alkynyl. In the embodiment, R 1 , R 2 and R 3 Each of these is independently, non-substituted C6~C 30 It is alkynyl. In the embodiment, R 1 , R 2 and R 3 Each of these independently substitutes C6~C 30 It is alkinyl.

[0371] In this embodiment, R 1 , R 2 and R 3 Each of these is independent of C4~C 30Heteroalkyl (e.g., C6~C) 30 It is a heteroalkyl group. In this embodiment, R 1 , R 2 and R 3 Each of these is independently, non-substituted C4~C 30 Heteroalkyl (e.g., unsubstituted C6~C) 30 It is a heteroalkyl group. In this embodiment, R 1 , R 2 and R 3 Each of these independently substitutes C4~C 30 Heteroalkyl (e.g., substituted C6~C) 30 It is a heteroalkyl group.

[0372] In this embodiment, B is independently an ionizable nitrogen-containing group.

[0373] In this embodiment, B is independently a permanently charged nitrogen-containing group.

[0374] In the embodiment, m is 2 and / or n is 2 or 3.

[0375] In this embodiment, m is 2 and n is 2.

[0376] In this embodiment, m is 2 and n is 3.

[0377] In this embodiment, B is independently, [ka] That is the case.

[0378] In this embodiment, B is independently, [ka] That is the case.

[0379] In this embodiment, B is independently, [ka] That is the case.

[0380] In this embodiment, B is independently, [ka] That is the case.

[0381] In this embodiment, B is [ka] That is the case.

[0382] In this embodiment, B is [ka] That is the case.

[0383] In this embodiment, B is [ka] That is the case.

[0384] In this embodiment, the compound is selected from the group consisting of compounds (1), (2), (3), (4), (6), (9), (17), (24), (26), (27), (33), and (35).

[0385] Formula (III) In the embodiment, the compound has the structure of formula (III): [ka] Having or a pharmaceutically acceptable salt thereof, R 1 , R 2 , R 3 , R 4 Each of , and B independently follows any combination of embodiments described herein.

[0386] In this embodiment, R 1 , R 2 , and R 3 Each of these is independent of C6~C 30Alkyl, C6~C 30 Alkenil, C6~C 30 Alkinyl, or C4~C 30 It is heteroalkyl; R 4 C2~C 10 It is alkylene; B is an independently ionizable nitrogen-containing group.

[0387] In this embodiment, R 1 , R 2 and R 3 Each of these is independent of C6~C 30 It is alkyl. In this embodiment, R 1 , R 2 and R 3 Each of these is independently, non-substituted C6~C 30 It is alkyl. In this embodiment, R 1 , R 2 and R 3 Each of these independently substitutes C6~C 30 It is alkyl.

[0388] In this embodiment, R 1 , R 2 and R 3 Each of these is independent of C6~C 30 It is an alkenyl. In this embodiment, R 1 , R 2 and R 3 Each of these is independently, non-substituted C6~C 30 It is an alkenyl. In this embodiment, R 1 , R 2 and R 3 Each of these independently substitutes C6~C 30 It is Alkenil.

[0389] In this embodiment, R 1 , R 2 and R 3 Each of these is independent of C6~C 30 It is alkynyl. In the embodiment, R 1 , R 2 and R 3 Each of these is independently, non-substituted C6~C 30It is alkynyl. In the embodiment, R 1 , R 2 and R 3 Each of these independently substitutes C6~C 30 It is alkinyl.

[0390] In this embodiment, R 1 , R 2 and R 3 Each of these is independently C4~C 30 Heteroalkyl (e.g., C6~C) 30 It is a heteroalkyl group. In this embodiment, R 1 , R 2 and R 3 Each of these is independently, non-substituted C4~C 30 Heteroalkyl (e.g., unsubstituted C6~C) 30 It is a heteroalkyl group. In this embodiment, R 1 , R 2 and R 3 Each of these independently substitutes C4~C 30 Heteroalkyl (e.g., substituted C6~C) 30 It is a heteroalkyl group.

[0391] In this embodiment, R 4 is unsubstituted C2~C 10 It is alkylene.

[0392] In this embodiment, R 4 This is the substitution C2~C 10 It is alkylene.

[0393] In this embodiment, R 4 It is -CH2CH2-.

[0394] In this embodiment, B is independently, [ka] That is the case.

[0395] In this embodiment, B is independently, [ka] That is the case.

[0396] In this embodiment, B is independently, [ka] That is the case.

[0397] In this embodiment, B is independently, [ka] That is the case.

[0398] In this embodiment, B is [ka] That is the case.

[0399] In this embodiment, the compound is compound (3).

[0400] Formula (IV) In the embodiment, the compound has the structure of formula (IV): [ka] Having or a pharmaceutically acceptable salt thereof, R 1 , R 2 , R 3 Each of m, n, and B independently follows any combination of embodiments described herein.

[0401] In this embodiment, R 1 , R 2 , and R 3 Each of these is independent of C6~C 30 Alkyl, C6~C 30 Alkenil, C6~C 30 Alkinyl, or C4~C 30 It is heteroalkyl; m is an integer between 2 and 10; n is an integer between 2 and 10; B is an independently ionizable nitrogen-containing group.

[0402] In this embodiment, R 1 , R 2 and R 3 Each of these is independent of C6~C 30 It is alkyl. In this embodiment, R 1 , R 2 and R 3 Each of these is independently, non-substituted C6~C 30 It is alkyl. In this embodiment, R 1 , R 2 and R 3 Each of these independently substitutes C6~C 30 It is alkyl.

[0403] In this embodiment, R 1 , R 2 and R 3 Each of these is independent of C6~C 30 It is an alkenyl. In this embodiment, R 1 , R 2 and R 3 Each of these is independently, non-substituted C6~C 30 It is an alkenyl. In this embodiment, R 1 , R 2 and R 3 Each of these independently substitutes C6~C 30 It is Alkenil.

[0404] In this embodiment, R 1 , R 2 and R 3 Each of these is independent of C6~C 30 It is alkynyl. In the embodiment, R 1 , R 2 and R 3 Each of these is independently, non-substituted C6~C 30 It is alkynyl. In the embodiment, R 1 , R 2 and R 3 Each of these independently substitutes C6~C 30 It is alkinyl.

[0405] In this embodiment, R 1 , R 2 and R3 Each of these is independent of C4~C 30 Heteroalkyl (e.g., C6~C) 30 It is a heteroalkyl group. In this embodiment, R 1 , R 2 and R 3 Each of these is independently, non-substituted C4~C 30 Heteroalkyl (e.g., unsubstituted C6~C) 30 It is a heteroalkyl group. In this embodiment, R 1 , R 2 and R 3 Each of these independently substitutes C4~C 30 Heteroalkyl (e.g., substituted C6~C) 30 It is a heteroalkyl group.

[0406] In this embodiment, R 1 , R 2 , and R 3 C4~C 30 It is heteroalkyl.

[0407] In the embodiment, m is 2 and / or n is 2 or 3.

[0408] In this embodiment, m is 2 and n is 2.

[0409] In this embodiment, m is 2 and n is 3.

[0410] In this embodiment, B is independently, [ka] That is the case.

[0411] In this embodiment, B is independently, [ka] That is the case.

[0412] In this embodiment, B is independently, [ka] That is the case.

[0413] In this embodiment, B is independently, [ka] That is the case.

[0414] In this embodiment, B is [ka] That is the case.

[0415] In the embodiment, the compound is compound (8) or (39).

[0416] Formula (V1) In the embodiment, the compound has the structure of formula (V1): [ka] Having or a pharmaceutically acceptable salt thereof, R 5 , L 3 Each of n, o, and B independently follows any combination of embodiments described herein.

[0417] In the embodiment, n is an integer of 2, 3, 4, 5, 6, or 7; B is an independently ionizable nitrogen-containing group.

[0418] In this embodiment, L 3 is OC(O), CO2, or (O)CO; o is an integer between 2 and 5; R 5 C4~C 24 It is alkyl.

[0419] In this embodiment, L 3 is OC(O), CO2, or (O)CO; o is an integer between 6 and 12; R 5 is a C1-C5 alkyl group. In the embodiment, L 3is OC(O), CO2, or (O)CO; o is an integer between 6 and 12; R 5 These are C1-C6 alkyl groups.

[0420] In this embodiment, B is independently, [ka] That is the case.

[0421] In this embodiment, B is independently, [ka] That is the case.

[0422] In this embodiment, B is independently, [ka] That is the case.

[0423] In this embodiment, B is independently, [ka] That is the case.

[0424] In this embodiment, B is [ka] That is the case.

[0425] In the embodiment, the compound is selected from the group consisting of compounds (11), (12), (13), (14), (22), (25), (28), (29), (30), (32), (34), (36), (38), (40), (112), (113), (114), (115), (116), (117), (126), (127), (128), and (129).

[0426] Formula (V) In the embodiment, the compound has the structure of formula (V): [ka] Having or a pharmaceutically acceptable salt thereof, R 5 , L 3 Each of n, o, and B independently follows any combination of embodiments described herein.

[0427] In the embodiment, n is an integer of 2, 3, or 4; B is an independently ionizable nitrogen-containing group.

[0428] In this embodiment, L 3 is OC(O), CO2, or (O)CO; o is an integer between 2 and 5; R 5 C4~C 24 It is alkyl.

[0429] In this embodiment, L 3 is OC(O), CO2, or (O)CO; o is an integer between 6 and 12; R 5 These are C1-C5 alkyl groups.

[0430] In this embodiment, B is independently, [ka] That is the case.

[0431] In this embodiment, B is independently, [ka] That is the case.

[0432] In this embodiment, B is independently, [ka] That is the case.

[0433] In this embodiment, B is independently, [ka] That is the case.

[0434] In this embodiment, B is [ka] That is the case.

[0435] In the embodiment, the compound is selected from the group consisting of compounds (11), (12), (13), (14), (22), (25), (28), (29), (30), (32), (34), (36), (38), (40), (114), and (116).

[0436] In this embodiment, the compound is selected from the group consisting of compounds (11), (12), (13), (14), (22), (25), (28), (29), (30), (32), (34), (36), (38), and (40).

[0437] Equation (VI) In the embodiment, the compound has the structure of formula (VI): [ka] Having or a pharmaceutically acceptable salt thereof, R 1 , R 2 , R 3 Each of , n, and B independently follows any combination of embodiments described herein.

[0438] In the embodiment, R 1 , R 2 , and R 3 Each of them independently contains a disulfide group C6-C 30 It is heteroalkyl; n is an integer of 2, 3, or 4; B is an independently ionizable nitrogen-containing group.

[0439] In this embodiment, B is independently, [ka] That is the case.

[0440] In this embodiment, B is independently, [ka] That is the case.

[0441] In this embodiment, B is independently, [ka] That is the case.

[0442] In this embodiment, B is independently, [ka] That is the case.

[0443] In this embodiment, B is [ka] That is the case.

[0444] In this embodiment, the compound is compound (21).

[0445] Formula (VII) In one embodiment, the cationic lipid is related to formula (VII) [ka] A cationic lipid having or a pharmaceutically acceptable salt thereof, wherein the formula X is independently either O or NH; R 1 , R 2 , and R 3 Each of these is independent of C4~C 30 Alkyl, C4~C 30 Alkenyl, C4~C 30 Alkinyl, or C4~C 30 It is heteroalkyl; Each m is an integer between 2 and 10; Each L 4 is a carbonyl, ester, or amide; Z is -(CH2) q1 -N-(CH2) q2 -where q1 and q2 are independently integers between 2 and 10; or Z is C6H3-Z 1 And here, Z 1 is -CH2(CH2) q The B portion is covalently bonded to a carbonyl, ester, or amide; q is an integer between 1 and 9; B is an independently ionizable nitrogen-containing group.

[0446] In this embodiment, R 1 , R 2 and R 3 Each of these is independent of C4~C 30 It is alkyl. In this embodiment, R 1 , R 2 and R 3 Each of these is independently, non-substituted C4~C 30 It is alkyl. In this embodiment, R 1 , R 2 and R 3 Each of these independently substitutes C4~C 30 It is alkyl.

[0447] In this embodiment, R 1 , R 2 and R 3 Each of these is independent of C4~C 30 It is an alkenyl. In this embodiment, R 1 , R 2 and R 3 Each of these is independently, non-substituted C4~C 30 It is an alkenyl. In this embodiment, R 1 , R 2 and R 3 Each of these independently substitutes C4~C 30 It is Alkenil.

[0448] In this embodiment, R 1 , R 2and R 3 Each of these is independent of C4~C 30 It is alkynyl. In the embodiment, R 1 , R 2 and R 3 Each of these is independently, non-substituted C4~C 30 It is alkynyl. In the embodiment, R 1 , R 2 and R 3 Each of these independently substitutes C4~C 30 It is alkinyl.

[0449] In this embodiment, R 1 , R 2 and R 3 Each of these is independent of C4~C 30 Heteroalkyl (e.g., C6~C) 30 It is a heteroalkyl group. In this embodiment, R 1 , R 2 and R 3 Each of these is independently, non-substituted C4~C 30 Heteroalkyl (e.g., unsubstituted C6~C) 30 It is a heteroalkyl group. In this embodiment, R 1 , R 2 and R 3 Each of these independently substitutes C4~C 30 Heteroalkyl (e.g., substituted C6~C) 30 It is a heteroalkyl group.

[0450] In this embodiment, L 4 It is a carbonyl group.

[0451] In this embodiment, L 4 is an ester. In this embodiment, L 4 is -OC(O)-. In the embodiment, L 4 is -(O)CO-.

[0452] In this embodiment, L 4 is an amide. In this embodiment, L 4 is -NHC(O)-. In the embodiment, L 4 It is -C(O)NH-.

[0453] In the embodiment, X is O; each L 4 is -C(O)O-; m is an integer of 2, 3, or 4; and / or q is 1.

[0454] In this embodiment, Z is -(CH2) q1 -N-(CH2) q2 - and q1 and q2 are both 2.

[0455] In this embodiment, Z is [ka] That is the case.

[0456] In this embodiment, B is independently, [ka] That is the case.

[0457] In this embodiment, B is independently, [ka] That is the case.

[0458] In this embodiment, B is independently, [ka] That is the case.

[0459] In this embodiment, B is independently, [ka] That is the case.

[0460] In this embodiment, B is [ka] That is the case.

[0461] In the embodiment, the compound is compound (16) or (31).

[0462] In one embodiment, the present invention is characterized by a method for delivering a composition comprising mRNA encoding a protein or polypeptide encapsulated within a liposome to the lung, intranasal cavity, or intramuscular muscle, wherein the liposome comprises one or more cationic lipids, optionally one or more non-cationic lipids, optionally one or more cholesterol-based lipids, and optionally one or more PEG-modified lipids, wherein at least one cationic lipid is a compound having a structure relating to formula (A2), formula (A1), or formula (A) as described herein, or a pharmaceutically acceptable salt thereof.

[0463] In one embodiment, the present invention is characterized by a method of delivering a composition containing nucleic acids encapsulated in liposomes to the lungs, nasal cavity, or intramuscular muscle, wherein the liposomes contain cationic lipids which are compounds having a structure relating to formula (A2), formula (A1), or formula (A) as described herein, or pharmaceutically acceptable salts thereof.

[0464] In one embodiment, the present invention is characterized by a method of intranasal delivery of a composition comprising mRNA encoding a protein or polypeptide encapsulated within a liposome, wherein the liposome comprises one or more cationic lipids, optionally one or more non-cationic lipids, optionally one or more cholesterol-based lipids, and optionally one or more PEG-modified lipids, wherein at least one cationic lipid is a compound having a structure relating to formula (A2), formula (A1), or formula (A) as described herein, or a pharmaceutically acceptable salt thereof, for example, compound 64, or a compound in Table 6, or a pharmaceutically acceptable salt thereof.

[0465] In one embodiment, the present invention is characterized by a method for intranasal delivery of a composition containing nucleic acids encapsulated in liposomes, wherein the liposomes contain cationic lipids which are compounds having a structure relating to formula (A2), formula (A1), or formula (A) as described herein, or pharmaceutically acceptable salts thereof, such as compound 64, or compounds in Table 6, or pharmaceutically acceptable salts thereof.

[0466] Intranasal administration includes administration via the nose, with or without simultaneous inhalation during administration. Such administration is typically carried out via contact of the composition with the nasal mucosa, turbinates, or sinuses. The pharmaceutical composition for administration may be applied in single or multiple doses. For example, one dose may be placed in each nostril during administration. For example, two-dose delivery can be used with the composition according to the present invention. A two-dose device contains two subdoses of a single dose, one subdose for administration to each nostril. Generally, the two subdoses reside in a single chamber, and the construction of the device allows for efficient delivery of one subdose at a time. Alternatively, a single-dose device may be used to administer the composition according to the present invention. The composition may be administered in doses of 1, 2, 3, 4 or more, such that the subject is given a first dose (which may be two doses or a single dose as described above), and then a second dose is administered at intervals of 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, or 30 days, or 4, 5, 6, 7, 8, 9, 10, 11, or 12 weeks, or 3, 4, 5, 6, 7, 8, 9, 10, 11, or 12 months, or within 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10 years or longer. An exemplary device for intranasal administration of the composition according to the present invention is a spray device. A suitable commercially available nasal spray device is Accuspray® (Becton Dickinson). The sprayer produces a fine mist or atomizer that is easily inhaled, as intended herein. Exemplary spray devices for intranasal use are devices whose performance does not depend on the pressure applied by the user. These devices are known as pressure threshold devices. The liquid is released from the nozzle only when a threshold pressure is applied. These devices make it easier to achieve sprays of typical droplet size. Pressure threshold devices suitable for use with the present invention are known in the art.

[0467] In a further embodiment, the present invention provides a pharmaceutical kit comprising an intranasal administration device described herein containing a formulation according to the present invention.

[0468] The present invention is not necessarily limited to the spray delivery of liquid formulations. Compositions according to the present invention may be administered in other forms, such as powder.

[0469] Compounds 41-64 and 90 In another embodiment, the cationic lipid is selected from the group of compounds 41-64 and 90, or pharmaceutically acceptable salts thereof.

[0470] [Table 30]

[0471] [Table 31]

[0472] [Table 32]

[0473] [Table 33]

[0474] [Table 34]

[0475] [Table 35]

[0476] [Table 36]

[0477] Compounds 91~111 In another embodiment, the cationic lipid is selected from the group of compounds 91 to 111, or pharmaceutically acceptable salts thereof.

[0478] [Table 37]

[0479] [Table 38]

[0480] [Table 39]

[0481] [Table 40]

[0482] [Table 41]

[0483] Compounds 112~117 In another embodiment, the cationic lipid is selected from the group of compounds 112-117 and 126-129, or pharmaceutically acceptable salts thereof.

[0484] [Table 42]

[0485] [Table 43]

[0486] [Table 44]

[0487] Compounds 118~125 In another embodiment, the cationic lipid is selected from the group of compounds 118-125 or their pharmaceutically acceptable salts.

[0488] [Table 45]

[0489] [Table 46]

[0490] [Table 47]

[0491] Formula (B) In another embodiment, the present invention relates to a cationic lipid of formula (B). [ka] or characterized by a pharmaceutically acceptable salt thereof, in the formula, Each n is independently either 0 or 1; X 1A Independently, O or NR 1A and; R 1A is H or C1-C6 alkyl, X 1B is a covalent bond, C(O), CH2CO2, or CH2C(O); X 2A and X 2B One of them is O, and the other is a covalent bond; X 3A and X 3B One of them is O, and the other is a covalent bond; X 4A and X 4B One of them is O, and the other is a covalent bond; R 1 Independently, L 1 -B 1 , C6~C 30 Alkyl, C6~C30 Alkenyl, or C6~C 30 It is alkinyl; R 2 Independently, L 2 -B 2 , C6~C 30 Alkyl, C6~C 30 Alkenyl, or C6~C 30 It is alkinyl; R 3 Independently, L 3 -B 3 , C6~C 30 Alkyl, C6~C 30 Alkenyl, or C6~C 30 It is alkinyl; R 4 Independently, L 4 -B 4 , C6~C 30 Alkyl, C6~C 30 Alkenyl, or C6~C 30 It is alkinyl; L 1 , L 2 , L 3 , and L 4 Each of these is independent of C1~C 30 Alkylene; C2~C 30 Alkenylene; or C2~C 30 It is alkynylene; B 1 B 2 B 3 , and B 4 Each of them is independently an ionizable nitrogen-containing group, Cationic lipids contain at least one ionizable nitrogen-containing group.

[0492] In another embodiment, the present invention is characterized by a method for delivering a composition comprising mRNA encoding a protein or polypeptide encapsulated within a liposome to the lung, intranasal cavity, or intramuscular muscle, wherein the liposome comprises one or more cationic lipids, optionally one or more non-cationic lipids, optionally one or more cholesterol-based lipids, and optionally one or more PEG-modified lipids, wherein at least one cationic lipid is a compound having a structure relating to formula (B) as described herein, or a pharmaceutically acceptable salt thereof.

[0493] In another embodiment, the present invention is characterized by a method of delivering a composition containing nucleic acids encapsulated in liposomes to the lungs, nasal cavity, or intramuscular muscle, wherein the liposomes contain cationic lipids which are compounds having the structure of formula (B) as described herein, or pharmaceutically acceptable salts thereof.

[0494] In the embodiment, the compound of formula (B) is selected from the group consisting of compounds (65) to (89) or pharmaceutically acceptable salts thereof.

[0495] [Table 48]

[0496] [Table 49]

[0497] [Table 50]

[0498] [Table 51]

[0499] [Table 52]

[0500] [Table 53]

[0501] [Table 54]

[0502] In this embodiment, the compound of formula (B) is selected from the group consisting of compounds (65), (66), (67), (70), (71), (74), (75), (77), (81), (84), (87), (88), and (89).

[0503] In this embodiment, the compound of formula (B) is selected from the group consisting of compounds (68), (69), (72), (73), (76), (78), (79), (80), (82), (83), (85), and (86).

[0504] In this embodiment, the compounds of formula (B) exclude compounds (68), (69), (72), (73), (76), (78), (79), (80), (82), (83), (85), and (86).

[0505] Compound of formula (B1): [ka] It is also provided, and in the formula, B1 is, [ka] and; L1 is defined as in equation (B); R2, R3, and R4 are straight-chain C8 alkylenes.

[0506] Compound of formula (B1): [ka] It is also provided, and in the formula, B1 is, [ka] and; L1 is a linear C5 alkylene; R2, R3, and R4 are defined for equation (B).

[0507] In the embodiment, the compound of formula (B) or (B1) is selected from the group consisting of compounds (118) to (125) or pharmaceutically acceptable salts thereof.

[0508] [Table 55]

[0509] [Table 56]

[0510] [Table 57]

[0511] [Table 58]

[0512] Exemplary Compounds Examples of exemplary compounds of the present invention include the cationic lipids listed in Table 1.

[0513] [Table 59]

[0514] [Table 60]

[0515] [Table 61]

[0516] Table 62

[0517] Table 63

[0518] Table 64

[0519] Table 65

[0520] Table 66

[0521] Table 67

[0522] Table 68

[0523] Table 69

[0524] In the embodiment, the cationic lipid is compound (1) or a pharmaceutically acceptable salt thereof. In the embodiment, the cationic lipid is compound (2) or a pharmaceutically acceptable salt thereof. In the embodiment, the cationic lipid is compound (3) or a pharmaceutically acceptable salt thereof. In the embodiment, the cationic lipid is compound (4) or a pharmaceutically acceptable salt thereof. In the embodiment, the cationic lipid is compound (6) or a pharmaceutically acceptable salt thereof. In the embodiment, the cationic lipid is compound (8) or a pharmaceutically acceptable salt thereof. In the embodiment, the cationic lipid is compound (9) or a pharmaceutically acceptable salt thereof.

[0525] In the embodiment, the cationic lipid is compound (11) or a pharmaceutically acceptable salt thereof. In the embodiment, the cationic lipid is compound (12) or a pharmaceutically acceptable salt thereof. In the embodiment, the cationic lipid is compound (13) or a pharmaceutically acceptable salt thereof. In the embodiment, the cationic lipid is compound (14) or a pharmaceutically acceptable salt thereof. In the embodiment, the cationic lipid is compound (16) or a pharmaceutically acceptable salt thereof. In the embodiment, the cationic lipid is compound (17) or a pharmaceutically acceptable salt thereof.

[0526] In the embodiment, the cationic lipid is compound (21) or a pharmaceutically acceptable salt thereof. In the embodiment, the cationic lipid is compound (22) or a pharmaceutically acceptable salt thereof. In the embodiment, the cationic lipid is compound (24) or a pharmaceutically acceptable salt thereof. In the embodiment, the cationic lipid is compound (25) or a pharmaceutically acceptable salt thereof. In the embodiment, the cationic lipid is compound (26) or a pharmaceutically acceptable salt thereof. In the embodiment, the cationic lipid is compound (27) or a pharmaceutically acceptable salt thereof. In the embodiment, the cationic lipid is compound (28) or a pharmaceutically acceptable salt thereof. In the embodiment, the cationic lipid is compound (29) or a pharmaceutically acceptable salt thereof. In the embodiment, the cationic lipid is compound (30) or a pharmaceutically acceptable salt thereof.

[0527] In the embodiment, the cationic lipid is compound (31) or a pharmaceutically acceptable salt thereof. In the embodiment, the cationic lipid is compound (32) or a pharmaceutically acceptable salt thereof. In the embodiment, the cationic lipid is compound (33) or a pharmaceutically acceptable salt thereof. In the embodiment, the cationic lipid is compound (34) or a pharmaceutically acceptable salt thereof. In the embodiment, the cationic lipid is compound (35) or a pharmaceutically acceptable salt thereof. In the embodiment, the cationic lipid is compound (36) or a pharmaceutically acceptable salt thereof. In the embodiment, the cationic lipid is compound (38) or a pharmaceutically acceptable salt thereof. In the embodiment, the cationic lipid is compound (39) or a pharmaceutically acceptable salt thereof. In the embodiment, the cationic lipid is compound (40) or a pharmaceutically acceptable salt thereof.

[0528] Exemplary compounds of the present invention include the cationic lipids listed in Table 2.

[0529] [Table 70]

[0530] [Table 71]

[0531] [Table 72]

[0532] [Table 73]

[0533] [Table 74]

[0534] [Table 75]

[0535] [Table 76]

[0536] In the embodiment, the cationic lipid is compound (41) or a pharmaceutically acceptable salt thereof. In the embodiment, the cationic lipid is compound (42) or a pharmaceutically acceptable salt thereof. In the embodiment, the cationic lipid is compound (43) or a pharmaceutically acceptable salt thereof. In the embodiment, the cationic lipid is compound (44) or a pharmaceutically acceptable salt thereof. In the embodiment, the cationic lipid is compound (45) or a pharmaceutically acceptable salt thereof. In the embodiment, the cationic lipid is compound (46) or a pharmaceutically acceptable salt thereof. In the embodiment, the cationic lipid is compound (47) or a pharmaceutically acceptable salt thereof. In the embodiment, the cationic lipid is compound (48) or a pharmaceutically acceptable salt thereof. In the embodiment, the cationic lipid is compound (49) or a pharmaceutically acceptable salt thereof. In the embodiment, the cationic lipid is compound (50) or a pharmaceutically acceptable salt thereof.

[0537] In the embodiment, the cationic lipid is compound (51) or a pharmaceutically acceptable salt thereof. In the embodiment, the cationic lipid is compound (52) or a pharmaceutically acceptable salt thereof. In the embodiment, the cationic lipid is compound (53) or a pharmaceutically acceptable salt thereof. In the embodiment, the cationic lipid is compound (54) or a pharmaceutically acceptable salt thereof. In the embodiment, the cationic lipid is compound (55) or a pharmaceutically acceptable salt thereof. In the embodiment, the cationic lipid is compound (56) or a pharmaceutically acceptable salt thereof. In the embodiment, the cationic lipid is compound (57) or a pharmaceutically acceptable salt thereof. In the embodiment, the cationic lipid is compound (58) or a pharmaceutically acceptable salt thereof. In the embodiment, the cationic lipid is compound (59) or a pharmaceutically acceptable salt thereof. In the embodiment, the cationic lipid is compound (60) or a pharmaceutically acceptable salt thereof.

[0538] In the embodiment, the cationic lipid is compound (61) or a pharmaceutically acceptable salt thereof. In the embodiment, the cationic lipid is compound (62) or a pharmaceutically acceptable salt thereof. In the embodiment, the cationic lipid is compound (63) or a pharmaceutically acceptable salt thereof. In the embodiment, the cationic lipid is compound (64) or a pharmaceutically acceptable salt thereof. In the embodiment, the cationic lipid is compound (90) or a pharmaceutically acceptable salt thereof.

[0539] Exemplary compounds of the present invention include the cationic lipids listed in Table 3.

[0540] [Table 77]

[0541] [Table 78]

[0542] [Table 79]

[0543] [Table 80]

[0544] [Table 81]

[0545] [Table 82]

[0546] [Table 83]

[0547] [Table 84]

[0548] In the embodiment, the cationic lipid is compound (65) or a pharmaceutically acceptable salt thereof. In the embodiment, the cationic lipid is compound (66) or a pharmaceutically acceptable salt thereof. In the embodiment, the cationic lipid is compound (67) or a pharmaceutically acceptable salt thereof. In the embodiment, the cationic lipid is compound (68) or a pharmaceutically acceptable salt thereof. In the embodiment, the cationic lipid is compound (69) or a pharmaceutically acceptable salt thereof. In the embodiment, the cationic lipid is compound (70) or a pharmaceutically acceptable salt thereof.

[0549] In the embodiment, the cationic lipid is compound (71) or a pharmaceutically acceptable salt thereof. In the embodiment, the cationic lipid is compound (72) or a pharmaceutically acceptable salt thereof. In the embodiment, the cationic lipid is compound (73) or a pharmaceutically acceptable salt thereof. In the embodiment, the cationic lipid is compound (74) or a pharmaceutically acceptable salt thereof. In the embodiment, the cationic lipid is compound (75) or a pharmaceutically acceptable salt thereof. In the embodiment, the cationic lipid is compound (76) or a pharmaceutically acceptable salt thereof. In the embodiment, the cationic lipid is compound (77) or a pharmaceutically acceptable salt thereof. In the embodiment, the cationic lipid is compound (78) or a pharmaceutically acceptable salt thereof. In the embodiment, the cationic lipid is compound (79) or a pharmaceutically acceptable salt thereof. In the embodiment, the cationic lipid is compound (80) or a pharmaceutically acceptable salt thereof.

[0550] In the embodiment, the cationic lipid is compound (81) or a pharmaceutically acceptable salt thereof. In the embodiment, the cationic lipid is compound (82) or a pharmaceutically acceptable salt thereof. In the embodiment, the cationic lipid is compound (83) or a pharmaceutically acceptable salt thereof. In the embodiment, the cationic lipid is compound (84) or a pharmaceutically acceptable salt thereof. In the embodiment, the cationic lipid is compound (85) or a pharmaceutically acceptable salt thereof. In the embodiment, the cationic lipid is compound (86) or a pharmaceutically acceptable salt thereof. In the embodiment, the cationic lipid is compound (87) or a pharmaceutically acceptable salt thereof. In the embodiment, the cationic lipid is compound (88) or a pharmaceutically acceptable salt thereof. In the embodiment, the cationic lipid is compound (89) or a pharmaceutically acceptable salt thereof.

[0551] Exemplary compounds of the present invention include the cationic lipids listed in Table 9.

[0552] [Table 85]

[0553] Table 86

[0554] Table 87

[0555] Table 88

[0556] Table 89

[0557] In the embodiment, the cationic lipid is compound (91) or a pharmaceutically acceptable salt thereof. In the embodiment, the cationic lipid is compound (92) or a pharmaceutically acceptable salt thereof. In the embodiment, the cationic lipid is compound (93) or a pharmaceutically acceptable salt thereof. In the embodiment, the cationic lipid is compound (94) or a pharmaceutically acceptable salt thereof. In the embodiment, the cationic lipid is compound (95) or a pharmaceutically acceptable salt thereof. In the embodiment, the cationic lipid is compound (96) or a pharmaceutically acceptable salt thereof. In the embodiment, the cationic lipid is compound (97) or a pharmaceutically acceptable salt thereof. In the embodiment, the cationic lipid is compound (98) or a pharmaceutically acceptable salt thereof. In the embodiment, the cationic lipid is compound (99) or a pharmaceutically acceptable salt thereof. In the embodiment, the cationic lipid is compound (100) or a pharmaceutically acceptable salt thereof. In the embodiment, the cationic lipid is compound (101) or a pharmaceutically acceptable salt thereof. In the embodiment, the cationic lipid is compound (102) or a pharmaceutically acceptable salt thereof. In the embodiment, the cationic lipid is compound (103) or a pharmaceutically acceptable salt thereof. In the embodiment, the cationic lipid is compound (104) or a pharmaceutically acceptable salt thereof. In the embodiment, the cationic lipid is compound (105) or a pharmaceutically acceptable salt thereof. In the embodiment, the cationic lipid is compound (106) or a pharmaceutically acceptable salt thereof. In the embodiment, the cationic lipid is compound (107) or a pharmaceutically acceptable salt thereof. In the embodiment, the cationic lipid is compound (108) or a pharmaceutically acceptable salt thereof. In the embodiment, the cationic lipid is compound (109) or a pharmaceutically acceptable salt thereof. In the embodiment, the cationic lipid is compound (110) or a pharmaceutically acceptable salt thereof. In the embodiment, the cationic lipid is compound (111) or a pharmaceutically acceptable salt thereof.

[0558] Exemplary compounds of the present invention include the cationic lipids listed in Table 10.

[0559] [Table 90]

[0560] [Table 91]

[0561] [Table 92]

[0562] In the embodiment, the cationic lipid is compound (112) or a pharmaceutically acceptable salt thereof. In the embodiment, the cationic lipid is compound (113) or a pharmaceutically acceptable salt thereof. In the embodiment, the cationic lipid is compound (114) or a pharmaceutically acceptable salt thereof. In the embodiment, the cationic lipid is compound (115) or a pharmaceutically acceptable salt thereof. In the embodiment, the cationic lipid is compound (116) or a pharmaceutically acceptable salt thereof. In the embodiment, the cationic lipid is compound (117) or a pharmaceutically acceptable salt thereof. In the embodiment, the cationic lipid is compound (126) or a pharmaceutically acceptable salt thereof. In the embodiment, the cationic lipid is compound (127) or a pharmaceutically acceptable salt thereof. In the embodiment, the cationic lipid is compound (128) or a pharmaceutically acceptable salt thereof. In the embodiment, the cationic lipid is compound (129) or a pharmaceutically acceptable salt thereof.

[0563] Exemplary compounds of the present invention include the cationic lipids listed in Table 11.

[0564] [Table 93]

[0565] [Table 94]

[0566] [Table 95]

[0567] In the embodiment, the cationic lipid is compound (118) or a pharmaceutically acceptable salt thereof. In the embodiment, the cationic lipid is compound (119) or a pharmaceutically acceptable salt thereof. In the embodiment, the cationic lipid is compound (120) or a pharmaceutically acceptable salt thereof. In the embodiment, the cationic lipid is compound (121) or a pharmaceutically acceptable salt thereof. In the embodiment, the cationic lipid is compound (122) or a pharmaceutically acceptable salt thereof. In the embodiment, the cationic lipid is compound (123) or a pharmaceutically acceptable salt thereof. In the embodiment, the cationic lipid is compound (124) or a pharmaceutically acceptable salt thereof. In the embodiment, the cationic lipid is compound (125) or a pharmaceutically acceptable salt thereof.

[0568] As further described herein, compositions comprising any compound of the present invention include compositions comprising one cationic lipid, one or more non-cationic lipids, one or more cholesterol-based lipids, and / or one or more PEG-modified lipids, as described in the embodiments above. In embodiments, the composition is lipid nanoparticles. In embodiments, one or more cationic lipids account for about 30 mol% to 60 mol% of the lipid nanoparticles. In embodiments, one or more non-cationic lipids account for 10 mol% to 50 mol% of the lipid nanoparticles. In embodiments, one or more PEG-modified lipids account for 1 mol% to 10 mol% of the lipid nanoparticles. In embodiments, cholesterol-based lipids account for 10 mol% to 50 mol% of the lipid nanoparticles. In embodiments, the lipid nanoparticles encapsulate mRNA encoding nucleic acids, optionally peptides or proteins. In embodiments, the lipid nanoparticles have an mRNA encapsulation rate of at least 50%. In embodiments, the lipid nanoparticles have an mRNA encapsulation rate of at least 55%. In embodiments, the lipid nanoparticles have an mRNA encapsulation rate of at least 60%. In the embodiment, the lipid nanoparticles have an mRNA encapsulation rate of at least 65%. In the embodiment, the lipid nanoparticles have an mRNA encapsulation rate of at least 70%. In the embodiment, the lipid nanoparticles have an mRNA encapsulation rate of at least 75%. In the embodiment, the lipid nanoparticles have an mRNA encapsulation rate of at least 80%. In the embodiment, the lipid nanoparticles have an mRNA encapsulation rate of at least 85%. In the embodiment, the lipid nanoparticles have an mRNA encapsulation rate of at least 90%. In the embodiment, the lipid nanoparticles have an mRNA encapsulation rate of at least 95%.

[0569] In the embodiments described above, one of the compositions is for use in therapy.

[0570] In the embodiments described above, any one of the compositions of the embodiments described above is for use in a method of treating or preventing a disease that is suitable for treatment or prevention with a peptide or protein encoded by mRNA, and which optionally is (a) a protein deficiency (which optionally occurs in the liver, lungs, brain, or muscles), (b) an autoimmune disease, (c) an infectious disease, or (d) cancer.

[0571] In the embodiments, the composition is optionally administered intravenously, intrathecally, intramuscularly, or by pulmonary delivery through the spraying or use of an inhaler (e.g., a metered-dose inhaler, a dry powder inhaler, or a soft mist inhaler).

[0572] Mucosal administration routes, such as oral and intranasal administration, can induce immune responses at both local and distal mucosal sites. Such administration routes can also produce systemic immune responses. Therefore, there is interest in using administration routes such as intranasal administration when administering the compositions of the present invention, particularly for the purpose of generating protective immunity. In embodiments, the methods and compositions of the present invention can induce mucosal immunity in a subject. In embodiments, the methods and compositions of the present invention can induce mucosal immunity at distal mucosal sites. In embodiments, the methods and compositions of the present invention can produce protective mucosal immunity. "Protective mucosal immunity" means immunity or an immune response to an infectious agent indicated by the subject, thereby preventing or improving the infection or reducing one or more of its symptoms. The compositions and methods described herein can induce an antibody response in a subject. In some embodiments, the antibody may be IgA. In some embodiments, the antibody may be IgG.

[0573] During infection or after vaccination in which a subject is exposed to a specific antigen, antigen-specific T cells are activated, proliferate, and then differentiate into effector cells. These effector cells help to eliminate the infection, but most of these cells subsequently die. The remaining memory cells can provide protection and / or an increased response if the subject later encounters the same antigen again. Therefore, it would be beneficial to identify methods and compositions that can induce an antigen-specific T cell response, for example, by inducing a CD4 memory T cell population. The compositions and methods described herein may enable a subject to produce a CD4 memory T cell population. In some embodiments, the compositions and methods described herein may induce a CD4 memory T cell population in a subject, optionally, the population is induced in the mucosa.

[0574] Compound Synthesis The compounds of the present invention described herein can be prepared according to methods known in the art, including those described herein.

[0575] nucleic acid The compounds of the present invention described herein can be used to prepare compositions useful for nucleic acid delivery.

[0576] Nucleic acid synthesis Nucleic acids according to the present invention can be synthesized according to any known method. For example, mRNA according to the present invention can be synthesized by in vitro transcription (IVT). Briefly, IVT is typically carried out using a linear or circular DNA template containing a promoter, a pool of ribonucleotide triphosphates, a buffer system that may contain DTT and magnesium ions, and a suitable RNA polymerase (e.g., T3, T7, mutant T7, or SP6 RNA polymerase), DNAse I, pyrophosphatase, and / or RNAse inhibitor. The exact conditions will vary depending on the specific application.

[0577] In some embodiments, the DNA template is transcribed in vitro for mRNA preparation according to the present invention. A suitable DNA template typically has a promoter for in vitro transcription, such as a T3, T7, mutant T7, or SP6 promoter, followed by the desired mRNA and a desired nucleotide sequence for terminal signaling.

[0578] The present invention allows for the determination of a desired mRNA sequence and its incorporation into a DNA template using standard methods. For example, starting from a desired amino acid sequence (e.g., an enzyme sequence), a virtual backtranslation is performed based on a degenerate genetic code. An optimization algorithm can then be used to select suitable codons. Typically, the G / C content can be optimized to achieve the highest possible G / C content, while simultaneously taking into account the frequency of tRNA according to the codon usage. The optimized RNA sequence can be established and displayed, for example, using a suitable display device, and compared to the original (wild-type) sequence. Secondary structure analysis can also be used to calculate stabilization and destabilization properties, or regions of RNA, respectively.

[0579] modified mRNA In some embodiments, the mRNA according to the present invention may be synthesized as unmodified or modified mRNA. Modified mRNA includes nucleotide modifications in RNA. Therefore, modified mRNA according to the present invention may include nucleotide modifications such as skeletal modifications, sugar modifications, or base modifications. In some embodiments, the mRNA may be derived from naturally occurring nucleotides and / or nucleotide analogs (modified nucleotides), including, but not limited to, purines (adenine (A), guanine (G)) or pyrimidines (thymine (T), cytosine (C), uracil (U)), and analogs or derivatives of modified nucleotides, purines, and pyrimidines, such as 1-methyl-adenine, 2-methyl-adenine, and 2-methylthio-N-6-iso Pentenyl adenine, N6-methyl adenine, N6-isopentenyl adenine, 2-thiocytosine, 3-methylcytosine, 4-acetylcytosine, 5-methylcytosine, 2,6-diaminopurine, 1-methylguanine, 2-methylguanine, 2,2-dimethylguanine, 7-methylguanine, inosine, 1-methylinosine, pseudouracil (5-uracil), dihydrouracil, 2-thiouracil, 4- Thio-uracil, 5-carboxymethylaminomethyl-2-thio-uracil, 5-(carboxyhydroxymethyl)-uracil, 5-fluoro-uracil, 5-bromo-uracil, 5-carboxymethylaminomethyl-uracil, 5-methyl-2-thio-uracil, 5-methyluracil, N-uracil-5-oxyacetate methyl ester, 5-methylaminomethyl-uracil, 5-methoxyaminomethyl-2-thio-uracil, 5'-meth It can be synthesized as xycarbonylmethyl-uracil, 5-methoxy-uracil, uracil-5-oxyacetate methyl ester, uracil-5-oxyacetate(v), 1-methyl-psoidouracil, quosin, β-D-mannosyl-quosin, weibtoxosin, and phosphoramidates, phosphorothioates, peptide nucleotides, methylphosphonates, 7-deazaguanosine, 5-methylcytosine, and inosine, among others.The preparation of such analogues is known to those skilled in the art from, for example, U.S. Patent Nos. 4,373,071, 4,401,796, 4,415,732, 4,458,066, 4,500,707, 4,668,777, 4,973,679, 5,047,524, 5,132,418, 5,153,319, U.S. Patent Nos. 5,262,530 and 5,700,642, and these disclosures are incorporated by reference as a whole.

[0580] Cationic lipids and nucleic acid pharmaceutical formulations In certain embodiments, the compounds of the present invention described herein, and pharmaceutical and liposome compositions comprising such lipids, can be used in formulations to facilitate the delivery of encapsulated material (e.g., one or more polynucleotides such as mRNA) to one or more target cells and the subsequent transfection of one or more target cells. For example, in certain embodiments, cationic lipids described herein (and liposome compositions comprising such lipids, etc.) are characterized by providing one or more release properties that give such compounds advantages over receptor-mediated endocytosis, clathrin-mediated and caveolae-mediated endocytosis, phagocytosis and 187ncapsule187 itosis, fusion, endosomal or lysosome disruption, and other similarly classified lipids.

[0581] According to the present invention, nucleic acids described herein, such as mRNA encoding proteins described herein (e.g., full-length, fragment, or partial protein), can be delivered via a delivery vehicle containing the compounds of the present invention described herein.

[0582] As used herein, the terms “delivery vehicle,” “transport vehicle,” and “nanoparticles” or their grammatical synonyms are interchangeable.

[0583] For example, the present invention provides a composition (e.g., a pharmaceutical composition) comprising a compound described herein and one or more polynucleotides. The composition (e.g., a pharmaceutical composition) may further comprise one or more cationic lipids, one or more non-cationic lipids, one or more cholesterol-based lipids and / or one or more PEG-modified lipids.

[0584] In certain embodiments, the compositions exhibit enhanced (e.g., increased) ability to transfect one or more target cells. Therefore, methods for transfecting one or more target cells are also provided herein. Such methods generally involve contacting one or more target cells with a cationic lipid and / or pharmaceutical composition disclosed herein (e.g., a liposomal formulation comprising a compound described herein that encapsulates one or more polynucleotides), thereby transfecting one or more target cells with the encapsulated material therein (e.g., one or more polynucleotides). As used herein, the terms “transfect” or “transfection” refer to the intracellular introduction of one or more encapsulation materials (e.g., nucleic acids and / or polynucleotides) into cells (e.g., target cells). The introduced polynucleotides may be maintained stably or transiently within the target cells. The term “transfection efficiency” refers to the relative amount of such encapsulation material (e.g., polynucleotides) taken up by, introduced into, and / or expressed by the target cells being transfected. In practice, transfection efficiency can be estimated by the amount of reporter polynucleotide product produced by target cells after transfection. In certain embodiments, the compounds and pharmaceutical compositions described herein exhibit high transfection efficiency, thereby improving the likelihood that an appropriate dose of the encapsulated material (e.g., one or more polynucleotides) will be delivered to the pathological site and subsequently expressed, while minimizing potential systemic adverse effects or toxicity associated with the compound or its encapsulated contents.

[0585] For example, after transfection of one or more target cells with polynucleotides encapsulated in one or more lipid nanoparticles containing a pharmaceutical or liposome composition disclosed herein, the production of products encoded by such polynucleotides (e.g., polypeptides or proteins) may be stimulated, and the ability of such target cells to express polynucleotides and produce, for example, the polypeptide or protein of interest may be enhanced. For example, transfection of target cells with one or more compounds or pharmaceutical compositions encapsulating mRNA will enhance (i.e., increase) the production of proteins or enzymes encoded by such mRNA.

[0586] Furthermore, the delivery media described herein (e.g., liposome delivery media) may be prepared to preferentially distribute to other target tissues, cells, or organs, such as the heart, lungs, kidneys, or spleen. In embodiments, the lipid nanoparticles of the present invention may be prepared to achieve enhanced delivery to target cells and tissues. For example, polynucleotides (e.g., mRNA) encapsulated in one or more of the compounds or pharmaceutical compositions and liposome compositions described herein can be delivered to and / or transfected to target cells or tissues. In some embodiments, the encapsulated polynucleotides (e.g., mRNA) may be expressed by target cells and produce (and possibly excreted) functional polypeptide products, thereby conferring beneficial properties to, for example, the target cells or tissues. Such encapsulated polynucleotides (e.g., mRNA) may encode, for example, hormones, enzymes, receptors, polypeptides, peptides, or other proteins of interest.

[0587] Liposome delivery medium In some embodiments, the composition is a suitable delivery vehicle. In some embodiments, the suitable delivery vehicle is a liposome delivery vehicle, such as lipid nanoparticles.

[0588] The terms "liposome delivery vehicle" and "liposome composition" are used interchangeably.

[0589] Enhancement of liposome compositions with one or more cationic lipids disclosed herein can be used as a means to improve (e.g., reduce) toxicity or to impart one or more desired properties (e.g., improved delivery of encapsulated polynucleotides to one or more target cells and / or reduced in vivo toxicity of the liposome composition) to such enhanced liposome compositions by other means. Accordingly, pharmaceutical compositions, particularly liposome compositions, comprising one or more cationic lipids disclosed herein are also intended.

[0590] Accordingly, in certain embodiments, the compounds of the present invention described herein may be used as components of a liposome composition to facilitate or enhance the delivery and release of an encapsulation material (e.g., one or more therapeutic agents) to one or more target cells (e.g., by permeating or fusing with the lipid membrane of such target cells).

[0591] As used herein, liposome delivery vehicles, such as lipid nanoparticles, are typically characterized as microscopic vesicles having an internal aqueous space isolated from an external medium by one or more bilayer membranes. The liposome bilayer is typically formed by amphiphilic molecules containing spatially separated hydrophilic and hydrophobic domains, such as synthetic or naturally occurring lipids (Lasic, Trends Biotechnol., 16:307-321, 1998). The liposome bilayer can also be formed by amphiphilic polymers and surfactants (e.g., polymerosomes, niosomes, etc.). In the context of the present invention, liposome delivery vehicles are typically used to deliver desired mRNA to target cells or tissues.

[0592] In certain embodiments, such compositions (e.g., liposome compositions) are loaded with or encapsulated with materials such as one or more biologically active polynucleotides (e.g., mRNA).

[0593] In embodiments, the composition (e.g., a pharmaceutical composition) comprises mRNA encoding a protein encapsulated within a liposome. In embodiments, the liposome comprises one or more cationic lipids, one or more non-cationic lipids, one or more cholesterol-based lipids, and one or more PEG-modified lipids, wherein at least one cationic lipid is a compound of the present invention as described herein. In embodiments, the composition comprises mRNA encoding a protein (e.g., any protein described herein). In embodiments, the composition comprises mRNA encoding a cystic fibrosis membrane conductance regulator (CFTR) protein. In embodiments, the composition comprises mRNA encoding an ornithine transcarbamylase (OTC) protein.

[0594] In the embodiments, the composition (e.g., a pharmaceutical composition) comprises nucleic acids encapsulated within liposomes, and the liposomes contain compounds described herein.

[0595] In embodiments, the nucleic acid is mRNA encoding a peptide or protein. In embodiments, the mRNA encodes a peptide or protein used for delivery to or treatment of the lung or lung cells of a target (for example, the mRNA encodes a cystic fibrosis membrane conductance regulator (CFTR) protein). In embodiments, the mRNA encodes a peptide or protein used for delivery to or treatment of the liver or hepatocytes of a target (for example, the mRNA encodes an ornithine transcarbamylase (OTC) protein). Further exemplary mRNAs are described herein.

[0596] In the embodiment, the liposome delivery vehicle (e.g., lipid nanoparticles) may have a net positive charge.

[0597] In the embodiment, the liposome delivery vehicle (e.g., lipid nanoparticles) may have a net negative charge.

[0598] In the embodiment, the liposome delivery vehicle (e.g., lipid nanoparticles) may have a net neutral charge.

[0599] In the embodiments, the lipid nanoparticles encapsulating nucleic acids (e.g., mRNA encoding peptides or proteins) comprise one or more compounds of the present invention as described herein.

[0600] For example, the amount of the compounds of the present invention described herein in a composition may be expressed as a percentage ("weight %) of the total dry weight of all lipids in the composition (e.g., the total dry weight of all lipids present in the liposome composition).

[0601] In embodiments of the pharmaceutical compositions described herein, the compounds of the present invention described herein are present in an amount of about 0.5% to about 30% by weight (for example, about 0.5% to about 20% by weight) of the total dry weight of all lipids present in the composition (e.g., liposome composition).

[0602] In embodiments, the compounds of the present invention described herein are present in amounts of about 1% to about 30% by weight, about 1% to about 20% by weight, about 1% to about 15% by weight, about 1% to about 10% by weight, or about 5% to about 25% by weight of the total dry weight of all lipids present in the composition (e.g., liposome composition). In embodiments, the compounds of the present invention described herein are present in amounts of about 0.5% to about 5% by weight, about 1% to about 10% by weight, about 5% to about 20% by weight, or about 10% to about 20% by weight of the total dry weight of all lipids present in the composition such as a liposome delivery vehicle.

[0603] In embodiments, the amount of the compounds of the present invention described herein is present in an amount of at least about 5% by weight, about 10% by weight, about 15% by weight, about 20% by weight, about 25% by weight, about 30% by weight, about 35% by weight, about 40% by weight, about 45% by weight, about 50% by weight, about 55% by weight, about 60% by weight, about 65% by weight, about 70% by weight, about 75% by weight, about 80% by weight, about 85% by weight, about 90% by weight, about 95% by weight, about 96% by weight, about 97% by weight, about 98% by weight, or about 99% by weight of the total dry weight of the total lipids in the composition (e.g., liposome composition).

[0604] In embodiments, the amount of the compounds of the present invention described herein is present in an amount of about 5% by weight, about 10% by weight, about 15% by weight, about 20% by weight, about 25% by weight or less of the total dry weight of the total lipids in the composition (e.g., liposome composition), and is present in an amount of about 30% by weight, about 35% by weight, about 40% by weight, about 45% by weight, about 50% by weight, about 55% by weight, about 60% by weight, about 65% by weight, about 70% by weight, about 75% by weight, about 80% by weight, about 85% by weight, about 90% by weight, about 95% by weight, about 96% by weight, about 97% by weight, about 98% by weight, or about 99% by weight or less.

[0605] In embodiments, the composition (e.g., a liposome delivery vehicle such as lipid nanoparticles) contains about 0.1% to about 20% by weight (e.g., about 0.1% to about 15% by weight) of the compound described herein. In embodiments, the delivery vehicle (e.g., a liposome delivery vehicle such as lipid nanoparticles) contains about 0.5% by weight, about 1% by weight, about 3% by weight, about 5% by weight, or about 10% by weight of the compound described herein. In embodiments, the delivery vehicle (e.g., a liposome delivery vehicle such as lipid nanoparticles) contains about 0.5% by weight, about 1% by weight, about 3% by weight, about 5% by weight, about 10% by weight, about 15% by weight, or up to about 20% by weight of the compound described herein. In embodiments, this percentage results in an improvement in beneficial effects (e.g., improved delivery to target tissues such as the liver or lungs).

[0606] The amount of the compounds of the present invention described herein in a composition may also be expressed as a percentage ("mol%) of the total molar amount of total lipids in the composition (for example, the total molar amount of all lipids present in the liposome delivery vehicle).

[0607] In embodiments of the pharmaceutical compositions described herein, the compounds of the present invention described herein are present in an amount of about 0.5 mol% to about 50 mol% (for example, about 0.5 mol% to about 20 mol%) of the total molar amount of all lipids present in the composition, such as a liposome delivery vehicle.

[0608] In embodiments, the compounds of the present invention described herein are present in amounts of about 0.5 mol% to about 5 mol%, about 1 mol% to about 10 mol%, about 5 mol% to about 20 mol%, about 10 mol% to about 20 mol%, about 15 mol% to about 30 mol%, about 20 mol% to about 35 mol%, about 25 mol% to about 40 mol%, about 30 mol% to about 45 mol%, about 35 mol% to about 50 mol%, about 40 mol% to about 55 mol%, or about 45 mol% to about 60 mol% of the total molar amount of all lipids present in a composition such as a liposome delivery vehicle. In embodiments, the compounds of the present invention as described herein are present in amounts of approximately 1 mol% to approximately 60 mol%, 1 mol% to approximately 50 mol%, 1 mol% to approximately 40 mol%, 1 mol% to approximately 30 mol%, approximately 1 mol% to approximately 20 mol%, approximately 1 mol% to approximately 15 mol%, approximately 1 mol% to approximately 10 mol%, approximately 5 mol% to approximately 55 mol%, approximately 5 mol% to approximately 45 mol%, approximately 5 mol% to approximately 35 mol%, or approximately 5 mol% to approximately 25 mol% of the total molar amount of all lipids present in a composition such as a liposome delivery medium.

[0609] In certain embodiments, the compounds of the present invention described herein may constitute about 0.1 mol% to about 50 mol%, or 0.5 mol% to about 50 mol%, or about 1 mol% to about 50 mol%, or about 5 mol% to about 50 mol%, or about 10 mol% to about 50 mol%, or about 15 mol% to about 50 mol%, or about 20 mol% to about 50 mol%, or about 25 mol% to about 50 mol%, or about 30 mol% to about 50 mol%, of the total amount of lipids in a composition (e.g., a liposome delivery vehicle).

[0610] In certain embodiments, the compounds of the present invention described herein may constitute more than 0.1 mol%, or about 0.5 mol%, or about 1 mol%, about 5 mol%, about 10 mol%, about 20 mol%, about 30 mol%, or about 40 mol% of the total amount of lipids in the lipid nanoparticles.

[0611] In certain embodiments, the compounds described may constitute less than about 60 mol%, or less than about 55 mol%, or less than about 50 mol%, or less than about 45 mol%, or less than about 40 mol%, or less than about 35 mol%, or less than about 30 mol%, or less than about 25 mol%, or less than about 10 mol%, or less than about 5 mol%, or less than about 1 mol% of the total amount of lipids in the composition (e.g., liposome delivery vehicle).

[0612] In embodiments, the amount of the compound of the present invention described herein is present in an amount that is at least about 5 mol%, about 10 mol%, about 15 mol%, about 20 mol%, about 25 mol%, about 30 mol%, about 35 mol%, about 40 mol%, about 45 mol%, about 50 mol%, about 55 mol%, about 60 mol%, about 65 mol%, about 70 mol%, about 75 mol%, about 80 mol%, about 85 mol%, about 90 mol%, about 95 mol%, about 96 mol%, about 97 mol%, about 98 mol%, or about 99 mol% of the total molar amount of total lipids in the composition (e.g., liposome composition).

[0613] In embodiments, the amount of the compounds of the present invention described herein is present in an amount that is about 5 mol%, about 10 mol%, about 15 mol%, about 20 mol%, about 25 mol%, about 30 mol%, about 35 mol%, about 40 mol%, about 45 mol%, about 50 mol%, about 55 mol%, about 60 mol%, about 65 mol%, about 70 mol%, about 75 mol%, about 80 mol%, about 85 mol%, about 90 mol%, about 95 mol%, about 96 mol%, about 97 mol%, about 98 mol%, or about 99 mol% or less of the total molar amount of total lipids in the composition (e.g., liposome composition).

[0614] In some embodiments, this percentage results in an improvement in beneficial effects (e.g., improved delivery to target tissues such as the liver or lungs).

[0615] In a typical embodiment, the composition of the present invention (e.g., a liposome composition) comprises one or more cationic lipids, one or more non-cationic lipids, one or more cholesterol-based lipids, and one or more PEG-modified lipids, wherein at least one cationic lipid is a compound of the present invention as described herein. For example, a composition suitable for carrying out the present invention comprises four lipid components: a compound of the present invention as described herein as a cationic lipid component, a non-cationic lipid, a cholesterol-based lipid, and a PEG-modified lipid. The non-cationic lipid may be DOPE or DEPE. The cholesterol-based lipid may be cholesterol. The PEG-modified lipid may be DMG-PEG2K.

[0616] In further embodiments, the pharmaceutical (e.g., liposome) composition comprises one or more PEG-modified lipids, non-cationic lipids, and cholesterol lipids. In other embodiments, such a pharmaceutical (e.g., liposome) composition comprises: one or more PEG-modified lipids; one or more non-cationic lipids; and one or more cholesterol lipids. In further embodiments, such a pharmaceutical (e.g., liposome) composition comprises: one or more PEG-modified lipids and one or more cholesterol lipids.

[0617] In the embodiments, the composition for encapsulating nucleic acids (e.g., mRNA encoding peptides or proteins) (e.g., lipid nanoparticles) comprises one or more compounds of the present invention as described herein and one or more lipids selected from the group consisting of cationic lipids, non-cationic lipids, and PEGylated lipids.

[0618] In embodiments, a composition (e.g., lipid nanoparticles) for encapsulating nucleic acids (e.g., mRNA encoding peptides or proteins) comprises one or more compounds of the present invention as described herein; one or more lipids selected from the group consisting of cationic lipids, non-cationic lipids, and PEGylated lipids, and further comprises cholesterol-based lipids. Typically, such a composition has four lipid components, comprising the compounds of the present invention as described herein as a cationic lipid component, a non-cationic lipid (e.g., DOPE), a cholesterol-based lipid (e.g., cholesterol), and a PEG-modified lipid (e.g., DMG-PEG2K).

[0619] In the embodiments, the lipid nanoparticles encapsulating nucleic acids (e.g., mRNA encoding peptides or proteins) include one or more compounds of the present invention as described herein, as well as one or more lipids selected from the group consisting of cationic lipids, non-cationic lipids, PEGylated lipids, and cholesterol-based lipids.

[0620] According to various embodiments, the cationic lipids, non-cationic lipids, and / or PEG-modified lipids constituting the lipid nanoparticles, and their relative molar ratios, are based on the characteristics of the selected lipids, the properties of the target cells, and the characteristics of the nucleic acids to be delivered. Further considerations include, for example, alkyl chain saturation, as well as the size, charge, pH, pKa, membrane fusion activity, and toxicity of the selected lipids. Therefore, the molar ratios can be adjusted as appropriate.

[0621] Synthesis of lipid nanoparticles In embodiments, the lipids described herein can be used to prepare lipid nanoparticles by methods known in the art. For example, a preferred method is the one described in International Publication No. 2018 / 089801 (which is incorporated herein by reference in its entirety).

[0622] One exemplary process for lipid nanoparticle formulations is Process A in International Publication No. 2018 / 089801 (see, for example, Example 1 and Figure 1 in International Publication No. 2018 / 089801). Process A ("A") relates to a conventional method of encapsulating mRNA by mixing mRNA with a lipid mixture without first pre-forming the lipids into lipid nanoparticles. In the exemplary process, an ethanol lipid solution and an aqueous buffer solution of mRNA are prepared separately. The solution of the lipid mixture (e.g., cationic lipids, helper lipids, zwitterionic lipids, PEG lipids) is prepared by dissolving the lipids in ethanol. The mRNA solution is prepared by dissolving mRNA in citrate buffer, resulting in mRNA at a concentration of 0.0833 mg / ml in citrate buffer at a pH of 4.5. These two solutions are then mixed using a pump system. In some examples, the two solutions are mixed using a gear pump system. In certain embodiments, the two solutions are mixed using a "T" junction (or "Y" junction). The mixture is then purified by diafiltration with a TFF process. The resulting formulation is concentrated and stored at 2-8°C until further use.

[0623] A second exemplary process for lipid nanoparticle formulations is Process B in International Publication No. 2018 / 089801 (see, for example, Example 2 and Figure 2 in International Publication No. 2018 / 089801). Process B ("B") refers to the process of encapsulating messenger RNA (mRNA) by mixing pre-formed lipid nanoparticles with mRNA. In Process B, a range of different conditions may be used, including various temperatures (i.e., whether or not the mixture is heated), buffers, and concentrations. In the exemplary process, lipids dissolved in ethanol and citrate buffer are mixed using a pump system. Instantaneous mixing of the two flows results in the formation of empty lipid nanoparticles, which is a self-assembly process. The resulting formulation mixture contains empty lipid nanoparticles in a citrate buffer containing alcohol. The formulation is then subjected to a TFF purification process, where buffer exchange takes place. Next, the resulting suspension of pre-formed empty lipid nanoparticles is mixed with mRNA using a pump system. For certain cationic lipids, heating the solution after mixing can increase the proportion of mRNA-containing lipid nanoparticles, potentially leading to a higher overall mRNA yield.

[0624] Cationic lipids In addition to any of the compounds of the present invention described herein, the composition may comprise one or more additional cationic lipids.

[0625] In some embodiments, liposomes may contain one or more additional cationic lipids. As used herein, the term “cationic lipid” refers to any of several lipid species that have a net positive charge at a selected pH, such as physiological pH. Several cationic lipids are described in the literature, and many are commercially available.

[0626] Suitable additional cationic lipids for use in the composition include those described in the literature.

[0627] Helper lipids A composition (e.g., a liposome composition) may also contain one or more helper lipids. Such helper lipids include noncationic lipids. As used herein, the term "noncationic lipid" refers to any neutral, zwitterionic, or anionic lipid. As used herein, the term "anionic lipid" refers to any of several lipid species that have a net negative charge at a selected pH, such as physiological pH. Noncationic lipids include, but are not limited to, distearoylphosphatidylcholine (DSPC), dioleoylphosphatidylcholine (DOPC), dipalmitoylphosphatidylcholine (DPPC), dioleoylphosphatidylglycerol (DOPG), dipalmitoylphosphatidylglycerol (DPPG), dioleoylphosphatidylethanolamine (DOPE), 1,2-diylcoyl-sn-glycero-3-phosphoethanolamine (DEPE), palmitoyloleoylphosphatidylcholine (POPC), and palmitoyloleoylphosphatidylethanolamine (PO Examples include PE), dioleoyl-phosphatidylethanolamine 4-(N-maleimidomethyl)-cyclohexane-l-carboxylate (DOPE-mal), dipalmitoylphosphatidylethanolamine (DPPE), dimyristoylphosphoethanolamine (DMPE), distearoyl-phosphatidylethanolamine (DSPE), 16-O-monomethylPE, 16-O-dimethylPE, 18-1-transPE, 1-stearoyl-2-oleoyl-phosphatidyethanolamine (phosphatidyethanolamine) (SOPE), or mixtures thereof. A noncationic or helper lipid suitable for carrying out the present invention is dioleoylphosphatidylethanolamine (DOPE). Alternatively, 1,2-dielcoyl-sn-glycero-3-phosphoethanolamine (DEPE) can be used as the noncationic or helper lipid.

[0628] In some embodiments, the noncationic lipid is a neutral lipid, i.e., a lipid that has no net charge under the conditions under which the composition is formulated and / or administered.

[0629] In some embodiments, noncationic lipids may be present in a molar ratio (mol%) of about 5% to about 90%, about 5% to about 70%, about 5% to about 50%, about 5% to about 40%, about 5% to about 30%, about 10% to about 70%, about 10% to about 50%, or about 10% to about 40% of the total lipids present in the composition. In some embodiments, the percentage of non-cationic lipids in liposomes may be greater than about 5 mol%, greater than about 10 mol%, greater than about 20 mol%, greater than about 30 mol%, or greater than about 40 mol%. In some embodiments, the percentage of total non-cationic lipids in liposomes may be greater than about 5 mol%, greater than about 10 mol%, greater than about 20 mol%, greater than about 30 mol%, or greater than about 40 mol%. In some embodiments, the percentage of non-cationic lipids in liposomes may be less than or equal to about 5 mol%, less than or equal to about 10 mol%, less than or equal to about 20 mol%, less than or equal to about 30 mol%, or less than or equal to about 40 mol%. In some embodiments, the percentage of total non-cationic lipids in liposomes may be less than or equal to about 5 mol%, less than or equal to about 10 mol%, less than or equal to about 20 mol%, less than or equal to about 30 mol%, or less than or equal to about 40 mol%.

[0630] In some embodiments, noncationic lipids may be present in the composition at a weight ratio (weight %) of about 5% to about 90%, about 5% to about 70%, about 5% to about 50%, about 5% to about 40%, about 5% to about 30%, about 10% to about 70%, about 10% to about 50%, or about 10% to about 40% of the total lipids present. In some embodiments, total noncationic lipids may be present in the composition at a weight ratio (weight %) of about 5% to about 90%, about 5% to about 70%, about 5% to about 50%, about 5% to about 40%, about 5% to about 30%, about 10% to about 70%, about 10% to about 50%, or about 10% to about 40% of the total lipids present. In some embodiments, the percentage of non-cationic lipids in liposomes may be greater than about 5% by weight, greater than about 10% by weight, greater than about 20% by weight, greater than about 30% by weight, or greater than about 40% by weight. In some embodiments, the percentage of total non-cationic lipids in liposomes may be greater than about 5% by weight, greater than about 10% by weight, greater than about 20% by weight, greater than about 30% by weight, or greater than about 40% by weight. In some embodiments, the percentage of non-cationic lipids in liposomes may be about 5% by weight or less, about 10% by weight or less, about 20% by weight or less, about 30% by weight or less, or about 40% by weight or less. In some embodiments, the percentage of total non-cationic lipids in liposomes may be about 5% by weight or less, about 10% by weight or less, about 20% by weight or less, about 30% by weight or less, or about 40% by weight or less.

[0631] Cholesterol-based lipids In some embodiments, the composition (e.g., liposome composition) comprises one or more cholesterol-based lipids. For example, a suitable cholesterol-based lipid for carrying out the present invention is cholesterol. Other suitable cholesterol-based lipids include, for example, DC-Chol (N,N-dimethyl-N-ethylcarboxamide cholesterol), 1,4-bis(3-N-oleylaminopropyl)piperazine (Gao, et al. Biochem. Biophys. Res. Comm. 179,280 (1991); Wolf et al. BioTechniques 23,139 (1997); U.S. Patent No. 5,744,335), or the following structure [ka] Examples include imidazole cholesterol esters (ICE) that have [specific properties].

[0632] In some embodiments, cholesterol-based lipids may be present in the liposome at a molar ratio (mol%) of about 1% to about 30% or about 5% to about 20% of the total lipids present. In some embodiments, the percentage of cholesterol-based lipids in lipid nanoparticles may be greater than about 5 mol%, greater than about 10 mol%, greater than about 20 mol%, greater than about 30 mol%, or greater than about 40 mol%. In some embodiments, the percentage of cholesterol-based lipids in lipid nanoparticles may be less than or equal to about 5 mol%, less than or equal to about 10 mol%, less than or equal to about 20 mol%, less than or equal to about 30 mol%, or less than or equal to about 40 mol%.

[0633] In some embodiments, cholesterol lipids may be present in the liposome at a weight ratio (wt%) of about 1% to about 30% or about 5% to about 20% of the total lipids present. In some embodiments, the percentage of cholesterol lipids in lipid nanoparticles may be greater than about 5% by weight, greater than about 10% by weight, greater than about 20% by weight, greater than about 30% by weight, or greater than about 40% by weight. In some embodiments, the percentage of cholesterol lipids in lipid nanoparticles may be less than or equal to about 5% by weight, less than or equal to about 10% by weight, less than or equal to about 20% by weight, less than or equal to about 30% by weight, or less than or equal to about 40% by weight.

[0634] PEGylated lipids In some embodiments, the composition (e.g., a liposome composition) comprises one or more further PEGylated lipids. A suitable PEGylated lipid for carrying out the present invention is 1,2-dimiristoyl-rac-glycero-3-methoxypolyethylene glycol-2000 (DMG-PEG2K).

[0635] For example, the use of derivatized lipids such as polyethylene glycol (PEG)-modified phospholipids and derivatized ceramides (PEG-CER) containing N-octanoyl-sphingosine-1-[succinyl(methoxypolyethylene glycol)-2000](C8 PEG-2000 ceramide) is also intended by the present invention in combination with one or more compounds of the present invention, and in some embodiments in combination with other lipids that together constitute liposomes. In some embodiments, particularly useful interchangeable lipids are those with shorter acyl chains (e.g., C8 PEG-2000 ceramide). 14 or C 18 It is a PEG-ceramide that has )

[0636] Further PEG-modified lipids intended (also referred to herein as PEGylated lipids; this term is treated as synonymous with PEG-modified lipids) include, but are not limited to, C6-C6. 20 Examples include polyethylene glycol chains up to 5 kDa in length, covalently bonded to lipids containing one or more alkyl chains of a certain length. In some embodiments, the PEG-modified or PEGylated lipids are PEGylated cholesterol or PEG-2K. The addition of such components may prevent complex aggregation and may also provide a means to increase circulating lifespan and increase delivery of the lipid-nucleic acid composition to target cells (Klibanov et al. (1990) FEBS Letters, 268(1):235-237), or they may be selected to be rapidly replaced from the formulation in vivo (see U.S. Patent No. 5,885,613).

[0637] Further PEG-modified phospholipids and derivatized lipids of the present invention may be present in a molar ratio (mol%) of about 0% to about 10%, about 0.5% to about 10%, about 1% to about 10%, about 2% to about 10%, or about 3% to about 5% of the total lipids present in the composition (e.g., liposome composition).

[0638] Pharmaceutical preparations and therapeutic use The compounds of the present invention described herein may be used to prepare compositions that facilitate or enhance the delivery and release of encapsulation materials (e.g., one or more therapeutic polynucleotides) to one or more target cells (e.g., by permeating or fusing with the lipid membranes of such target cells) (e.g., for constructing liposome compositions).

[0639] For example, if a liposome composition (e.g., lipid nanoparticles) contains one or more of the compounds disclosed herein or is concentrated with one or more of those compounds, a phase transition in the lipid bilayer of one or more target cells may facilitate the delivery of an encapsulation material (e.g., one or more therapeutic polynucleotides encapsulated in lipid nanoparticles) to one or more target cells.

[0640] Similarly, in certain embodiments, the compounds of the present invention described herein may be used to prepare liposome vehicles characterized by reduced in vivo toxicity. In certain embodiments, the reduced toxicity is a function of the high transfection efficiency associated with the compositions disclosed herein, meaning that the desired therapeutic response or outcome can be achieved even when the amount of such composition is reduced and administered to a subject.

[0641] Thus, pharmaceutical formulations comprising the compounds and nucleic acids described herein, provided by the present invention, can be used for a variety of therapeutic purposes. To facilitate in vivo delivery of nucleic acids, the compounds and nucleic acids described herein can be formulated in combination with one or more additional pharmaceutical carriers, targeted ligands, or stabilizing reagents. In some embodiments, the compounds described herein can be formulated via a pre-mixed lipid solution. In other embodiments, compositions comprising the compounds described herein can be formulated using post-insertion techniques for nanoparticles into lipid membranes. For drug formulation and administration techniques, refer to “Remington's Pharmaceutical Sciences,” Mack Publishing Co., Easton, Pa., latest edition.

[0642] Preferred routes of administration include, for example, intrapulmonary or intra-intestinal administration including oral, rectal, vaginal, mucosal, intratracheal, or inhalation; and parenteral delivery including intradermal, transdermal (topical), intramuscular, subcutaneous, intrathecal injection, and intrathecal, direct intraventricular, intravenous, intraperitoneal, or intranasal delivery. In detailed embodiments, intramuscular administration is to muscles selected from the group consisting of skeletal muscle, smooth muscle, and cardiac muscle. In some embodiments, administration results in delivery of nucleic acids to muscle cells. In some embodiments, administration results in delivery of nucleic acids to hepatocytes (i.e., liver cells).

[0643] A common route for administering the liposomal composition of the present invention may be intravenous delivery, particularly when treating metabolic disorders, especially those affecting the liver (e.g., ornithine transcarbamylase (OTC) deficiency). Alternatively, depending on the disease or disorder being treated, the liposomal composition may be administered via pulmonary delivery (e.g., for the treatment of cystic fibrosis). For vaccination, the liposomal composition of the present invention is typically administered intramuscularly. Diseases or disorders affecting the eye may be treated by intravitreous administration of the liposomal composition of the present invention.

[0644] Alternatively, or in addition, the pharmaceutical formulations of the present invention may be administered topically rather than systemically, for example, by direct injection of the pharmaceutical formulation into the targeted tissue (e.g., in a sustained-release formulation). Topical delivery may be achieved in a variety of ways depending on the target tissue. Exemplary tissues to which delivered mRNA may be delivered and / or expressed include, but are not limited to, the liver, kidneys, heart, spleen, serum, brain, skeletal muscle, lymph nodes, skin, and / or cerebrospinal fluid. In embodiments, the liver is the targeted tissue. For example, an aerosol containing the composition of the present invention may be inhaled (for nasal, tracheal, or bronchial delivery); the composition of the present invention may be injected, for example, into a site of injury, symptom onset of disease, or pain; the composition may be provided in lozenges for oral, tracheal, or esophageal application; it may be supplied in liquid, tablet, or capsule form for gastric or intestinal administration; it may be supplied in suppository form for rectal or vaginal application; or even in the eyes, it may be delivered using cream, eye drops, or even injection.

[0645] The compositions described herein may include mRNA encoding peptides (e.g., polypeptides such as proteins) that are described herein.

[0646] In this embodiment, mRNA encodes a polypeptide.

[0647] In this embodiment, mRNA encodes a protein.

[0648] Exemplary peptides encoded by mRNA (e.g., exemplary proteins encoded by mRNA) are described herein.

[0649] The present invention provides a method for delivering a composition having a full-length mRNA molecule encoding a target peptide or protein for use in the treatment of a target, such as a human target, or cells of a human target, or cells that are treated and delivered to a human target.

[0650] In certain embodiments, the present invention provides a method for producing a therapeutic composition having full-length mRNA encoding a peptide or protein for use in the delivery of a vaccine or in the treatment of a target or target cells. For example, in certain embodiments, the present invention provides a method for producing a therapeutic composition having full-length mRNA encoding an antigen from an infectious agent such as a virus.

[0651] In certain embodiments, the present invention provides a method for producing a therapeutic composition having full-length mRNA encoding an antigen associated with or identified from cancer cells of a target. In certain embodiments, the present invention provides a method for producing a therapeutic composition having full-length mRNA encoding an antigen determined from the cancer cells of the target itself, i.e., a method for providing a personalized cancer vaccine.

[0652] In the embodiment, the composition contains mRNA encoding a cystic fibrosis membrane conductance regulatory factor (CFTR) protein.

[0653] In the embodiment, the composition contains mRNA encoding ornithine transcarbamylase (OTC) protein.

[0654] Delivery method The delivery routes used in the methods of the present invention enable non-invasive self-administration of the compounds of the present invention. In some embodiments, the methods involve intratracheal or intrapulmonary administration by aerosol, spray therapy, or intravenous infusion of a composition containing mRNA encoding a therapeutic protein in a lipid carrier medium, as described above. In some embodiments, the protein is encapsulated in liposomes. In some embodiments, the liposomes contain lipids that are the compounds of the present invention. Where used herein, administration of the compounds of the present invention includes administration of a composition containing the compounds of the present invention.

[0655] While local cells and tissues of the lung are potential targets that can function as biological depots or reservoirs for the production and secretion of mRNA-encoded proteins, the applicants have discovered that by administering the compounds of the present invention to the lungs via aerosolization, spraying, or infusion, non-secretory proteins can be distributed even outside of lung cells. Although we do not wish to be constrained by any particular theory, the nanoparticle compositions of the present invention cross the airway-blood barrier, so it is intended that intact nanoparticles will be transferred to non-lung cells and tissues, such as the heart, liver, and spleen, and that the encoded proteins will be produced in those non-lung tissues. Thus, the usefulness of the compounds and methods of the present invention extends beyond the production of therapeutic proteins in lung cells and lung tissues and can be used for delivery to non-lung target cells and / or tissues. These are useful in the management and treatment of numerous diseases, particularly peripheral diseases caused by deficiencies of both secretory and non-secretory proteins and / or enzymes (e.g., one or more lysosomal storage disorders). In certain embodiments, when the compounds of the present invention are used in the methods of the present invention, the distribution of mRNA-encapsulated nanoparticles and the production of the encoded protein occur in the liver, spleen, heart, and / or other non-lung cells. For example, administration of the compounds of the present invention by aerosolization, spraying, or injection into the lungs would result in the composition itself and its protein products (e.g., functional β-galactosidase protein) being detectable in both local cells and tissues of the lungs and in peripheral target cells, tissues, and organs as a result of the migration of mRNA and delivery vehicles to non-lung cells.

[0656] In certain embodiments, the compounds of the present invention may be used in the methods of the present invention to specifically target peripheral cells or tissues. After pulmonary delivery, the compounds of the present invention are intended to cross the pulmonary airway blood barrier and be distributed to cells other than local lung cells. Accordingly, the compounds disclosed herein are administered to a target by an intrapulmonary route using various approaches known to those skilled in the art (e.g., by inhalation) and distributed to both local target cells and tissues of the lung, as well as peripheral non-lung cells and tissues (e.g., liver, spleen, kidney, heart, skeletal muscle cells, lymph nodes, brain, cerebrospinal fluid, and plasma). As a result, both local cells of the lung and peripheral non-lung cells can function as biological reservoirs or depots capable of producing and / or secreting translation products encoded by one or more polynucleotides. Accordingly, the present invention is not limited to the treatment of lung diseases or conditions and can be used as a non-invasive means to promote the delivery of polynucleotides or the production of enzymes and proteins encoded therein in terminal organs, tissues, and cells (e.g., hepatocytes) which would otherwise only be achieved by systemic administration. Examples of peripheral non-pulmonary cells include, but are not limited to, hepatocytes, epithelial cells, hematopoietic cells, epithelial cells, endothelial cells, osteocytes, stem cells, mesenchymal cells, nerve cells, cardiac cells, adipocytes, vascular smooth muscle cells, cardiomyocytes, skeletal muscle cells, beta cells, pituitary cells, synovial-lining cells, ovarian cells, testicular cells, fibroblasts, B cells, T cells, reticulocytes, leukocytes, granulocytes, and tumor cells.

[0657] After administration of the composition to the target, the mRNA-encoded protein product (e.g., functional protein or enzyme) is detectable in peripheral target tissue for at least approximately 1 to 7 days or longer after administration of the compound to the target. The amount of protein product required to achieve a therapeutic effect will vary depending on the disease state under treatment, the encoded protein, and the patient's condition. For example, the protein product should be at least 0.025 to 1.5 μg / ml (e.g., at least 0.050 μg / ml, at least 0.075 μg / ml, at least 0.1 μg / ml, at least 0.2 μg / ml, at least 0.3 μg / ml, at least 0.4 μg / ml, at least 0.5 μg / ml, at least 0.6 μg / ml, at least 0.7 μg / ml, at least 0.8 μg / ml, at least 0.9 μg / ml, at least 1.0 μg / ml, and at least... The compound may be detectable in peripheral target tissues for at least 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 35, 40, 45 days or more after administration of the compound to the subject, at concentrations of 1.1 μg / ml, at least 1.2 μg / ml, at least 1.3 μg / ml, at least 1.4 μg / ml, or at least 1.5 μg / ml (e.g., therapeutic concentrations).

[0658] Nucleic acids have been demonstrated to be delivered to the lungs by intratracheal administration of a liquid suspension of the compound and by inhalation or reference of an aerosol mist generated by a liquid sprayer, as described in U.S. Patent No. 5,780,014 incorporated herein.

[0659] In certain embodiments, the compounds of the present invention may be formulated to be delivered to a subject as an aerosol or as a particulate liquid or solid before or at the time of administration. Such compounds may be administered with the help of one or more suitable devices for administering such solid or liquid particulate compositions (e.g., aerosolized aqueous solutions or suspensions) to generate particles that can be easily breathed or inhaled by the subject. In some embodiments, such devices (e.g., metered-dose inhalers, jet sprayers, ultrasonic sprayers, dry powder inhalers, propellant-type inhalers or ventilators) facilitate the administration of a predetermined mass, volume or dose (e.g., about 0.5 mg / kg of mRNA per dose) of the composition to the subject. For example, in certain embodiments, the compounds of the present invention are administered to a subject using a metered-dose inhaler containing a suspension or solution of the compound and a suitable propellant. In certain embodiments, the compounds of the present invention may be formulated as particulate powders intended for inhalation (e.g., breathable dry particles). In certain embodiments, the compositions of the present invention, formulated as breathable particles, are breathable by the subject or delivered using a suitable device (e.g., average D50 or D90 particle size of about 500 μm, 400 μm, 300 μm, 250 μm, 200 μm, 150 μm, 100 μm, 75 μm, 50 μm, 25 μm, 20 μm, 15 μm, 12.5 μm, 10 μm, 5 μm, or 2.5 μm or less). In yet other embodiments, the compounds of the present invention are formulated to contain one or more lung surfactants (e.g., lamellar bodies).In some embodiments, the compound of the present invention is present in amounts of at least 0.05 mg / kg, at least 0.1 mg / kg, at least 0.5 mg / kg, at least 1.0 mg / kg, at least 2.0 mg / kg, at least 3.0 mg / kg, at least 4.0 mg / kg, at least 5.0 mg / kg, at least 6.0 mg / kg, at least 7.0 mg / kg, at least 8.0 mg / kg, at least 9.0 mg / kg, at least 10 mg / kg, at least 15 mg / kg, at least 20 mg / kg, and at least The drug is administered to the subject in a single dose such that a concentration of 25 mg / kg, at least 30 mg / kg, at least 35 mg / kg, at least 40 mg / kg, at least 45 mg / kg, at least 50 mg / kg, at least 55 mg / kg, at least 60 mg / kg, at least 65 mg / kg, at least 70 mg / kg, at least 75 mg / kg, at least 80 mg / kg, at least 85 mg / kg, at least 90 mg / kg, at least 95 mg / kg, or at least 100 mg / kg body weight is administered. In some embodiments, the compounds of the present invention are administered to a subject in one or more doses, such that a total amount of mRNA of at least 0.1 mg, at least 0.5 mg, at least 1.0 mg, at least 2.0 mg, at least 3.0 mg, at least 4.0 mg, at least 5.0 mg, at least 6.0 mg, at least 7.0 mg, at least 8.0 mg, at least 9.0 mg, at least 10 mg, at least 15 mg, at least 20 mg, at least 25 mg, at least 30 mg, at least 35 mg, at least 40 mg, at least 45 mg, at least 50 mg, at least 55 mg, at least 60 mg, at least 65 mg, at least 70 mg, at least 75 mg, at least 80 mg, at least 85 mg, at least 90 mg, at least 95 mg, or at least 100 mg is administered. [Examples]

[0660] Abbreviation DCM = Dichloromethane DIPEA = N,N-diisopropylethylamine DM = Desalted DMAP = 4-dimethylaminopyridine EDC-HCl=N-(3-dimethylaminopropyl)-N'-ethylcarbodiimide hydrochloride EDCI = 1-ethyl-3-(3-dimethylaminopropyl)carbodiimide ELSD = Evaporative Light Scattering Detection alkyl = ethyl acetate h=time rt=room temperature SM = Starting materials TLC = Thin-Layer Calculation

[0661] Example 1. Synthesis of JC-TL1-10D-E6-2 (compound (13)) Synthesis Scheme A [ka] Intermediate [A-3]: [ka] A stirred solution of hexane-1,6-diol [A-1] (10 g, 0.085 mol) and triethylamine (42.9 g, 0.45 mol) in dichloromethane (150 mL) was cooled to 0°C. 4-(dimethylamino)pyridine-1-ium (1.03 g, 0.0085 mol), followed by propionyl chloride [A-2] (3.93 g, 0.042 mol), was added, and the reaction mixture was stirred at room temperature for 16 hours. The progress of the reaction was monitored by TLC (SM was consumed). The reaction mixture was quenched to pH 7 with cold saturated NaHCO3 and extracted with DCM (3 × 100 mL). The resulting organic layer was dried over Na2SO4 and concentrated under reduced pressure. The crude product was purified by flash column chromatography (SiO2: 0-20% ethyl acetate in hexane) to obtain the desired 6-hydroxyhexylpropionate [A-3] (4.5 g, yield 30%) as a pale yellow oil. 1H NMR(400MHz,CDCl3):δ 4.07(t,J=5.6Hz,2H),3.64(br,2H),2.32(q,J=7.6Hz,2H),1.67-1.60(m,2H),1.58-1.54(m,2H),1.44-1.37(m,4H),1.13(t,J=7.6Hz,3H).

[0662] Intermediate [A-5]: [ka] A stirred solution of 6-hydroxyhexylpropionate [A-3] (4.5 g, 0.026 mol) and citric acid [A-4] (1.25 g, 0.0065 mol) in dichloromethane (100 mL) was cooled to 0°C, and EDC.HCl (4.95 g, 0.026 mol), followed by 4-(dimethylamino)pyridine-1-ium (0.78 g, 0.0065 mol), was added and the mixture was stirred at room temperature for 48 hours. The progress of the reaction was monitored by TLC (SM was consumed). Water (50 mL) was added to the reaction mixture and extracted with DCM (3 × 100 mL). The resulting organic layer was dried over Na2SO4 and concentrated under reduced pressure to obtain the crude product, which was purified by flash column chromatography (0-30% ethyl acetate in n-hexane) to obtain the pure product tris(6-(propionyloxy)hexyl)2-hydroxypropane-1,2,3-tricarboxylate[A-5] (0.57 g, yield 13%) as a pale yellow liquid. 1 H NMR(400MHz,CDCl3):δ 4.21(t,J=6.8Hz,2H),4.12-4.04(m,10H),2.88(d,J=15.6Hz,2H),2.79(d,J=15.6Hz,2H),2.32(q ,J=7.6Hz,6H),1.71-1.67(m,2H),1.64-1.62(m,10H),1.38-1.36(m,12H),1.13(t,J=7.8Hz,9H).

[0663] ELSD analysis: Purity 98.66%, calculated value C 33 H 56 O 13=660.37, measured value =661.10 (m / z, M+H + ).

[0664] JC-TL1-10D-E6-2 (compound (13)) [ka] A stirred solution of tris(6-(propionyloxy)hexyl)2-hydroxypropane-1,2,3-tricarboxylate[A-5] (0.57 g, 0.00086 mol) and 3-(dimethylamino)propanoic acid[6] (0.404 g, 0.00345 mol) in dichloromethane (15 mL) was cooled to 0°C, and EDC.HCl (0.662 g, 0.00345 mol), followed by 4-(dimethylamino)pyridine-1-ium (0.105 g, 0.00086 mol), was added and the mixture was stirred at room temperature for 48 hours. The progress of the reaction was monitored by TLC (SM was consumed). Water (20 mL) was added to the reaction mixture and extracted with DCM (3 × 50 mL). The obtained organic layer was dried over Na2SO4 and concentrated under reduced pressure to obtain the crude product, which was purified by flash column chromatography (0-100% ethyl acetate in n-hexane) to obtain the pure product tris(6-(propionyloxy)hexyl)2-((3-(dimethylamino)propanoyl)oxy)propane-1,2,3-tricarboxylate [JC-TL1-10D-E6-2] (compound (13)) (0.115 g, yield 17.4%) as a yellow liquid. 1 H NMR(400MHz,DMSO-d6):δ 4.05-3.97(m,12H),3.16(d,J=15.2Hz,2H),3.08(d,J=15.2Hz,2H),2.46-2.37(m,4H),2.28( q,J=7.6Hz,6H),2.10(s,6H),1.56-1.53(m,12H),1.31-1.30(m,12H),1.01(t,J=6.8Hz,9H).

[0665] ELSD analysis: Purity 99.66%, calculated value C 38 H 65 NO 14=759.44, measured value =760.50 (m / z, M+H + ).

[0666] Example 2. Synthesis protocol for M1266-J03983-009 (JC-TL1-12D-E4-6) (Compound (12)) Synthesis scheme B [ka] Intermediate [B-3]: [ka] A stirred solution of butane-1,4-diol [B-1] (10 g, 0.111 mol) and heptanoic acid [B-2] (14.4 g, 0.111 mol) in dichloromethane (200 mL) was cooled to 0°C, and EDC.HCl (25.5 g, 0.133 mol), followed by 4-(dimethylamino)pyridine-1-ium (1.35 g, 0.011 mol), was added and the mixture was stirred at room temperature for 48 hours. The progress of the reaction was monitored by TLC (SM was consumed). Water (200 mL) was added to the reaction mixture and extracted with DCM (3 × 300 mL). The resulting organic layer was dried over Na₂SO₄ and concentrated under reduced pressure to obtain the crude product, which was purified by flash column chromatography (0-20% ethyl acetate in n-hexane) to obtain the pure product 4-hydroxybutylheptanoate [B-3] (6.2 g, yield 27%) as a yellow liquid.

[0667] result: 1 H NMR (400MHz, CDCl3): δ 4.09(t,J=6.0Hz,2H),3.69-3.65(m,2H),2.28(t,J=7.6Hz,2H),1.73-1.59(m,6H),1.28(br,6H),0.87-0.85(m,3H).

[0668] Intermediate [B-5]: [ka] A stirred solution of 4-hydroxybutylheptanoate [B-3] (6.18 g, 0.0306 mol) and citric acid [B-4] (1.47 g, 0.00765 mol) in dichloromethane (150 mL) was cooled to 0°C, and EDC.HCl (5.84 g, 0.0306 mol), followed by 4-(dimethylamino)pyridine-1-ium (0.9 g, 0.00765 mol), was added and the mixture was stirred at room temperature for 48 hours. The progress of the reaction was monitored by TLC (SM was consumed). Water (100 mL) was added to the reaction mixture and extracted with DCM (3 × 300 mL). The resulting organic layer was dried over Na2SO4 and concentrated under reduced pressure to obtain the crude product, which was purified by flash column chromatography (0-30% ethyl acetate in n-hexane) to obtain the pure product tris(4-(heptanoyloxy)butyl)2-hydroxypropane-1,2,3-tricarboxylate[B-5] (1.53 g, yield 27%) as a colorless liquid. 1 H NMR(400MHz,CDCl3):δ 4.25(t,J=6.0Hz,2H),4.14-4.07(m,10H),2.88(d,J=15.6Hz,2H),2.80(d,J=15.6Hz,2H),2.29(t ,J=7.6Hz,6H),1.85-1.68(m,12H),1.64-1.56(m,6H),1.34-1.22(m,18H),0.88(t,J=7.8Hz,9H).

[0669] ELSD analysis: Purity 98.99%, calculated value C 39 H 68 O 13 =744.46, measured value =745.25 (m / z, M+H + ).

[0670] JC-TL1-12D-E4-6 (compound (12)) [ka] A stirred solution of tris(4-(heptanoyloxy)butyl)2-hydroxypropane-1,2,3-tricarboxylate[5] (1.53 g, 0.00205 mol) and 3-(dimethylamino)propanoic acid[B-6] (0.962 g, 0.00822 mol) in dichloromethane (30 mL) was cooled to 0°C, and EDC.HCl (1.56 g, 0.00822 mol), followed by 4-(dimethylamino)pyridine-1-ium (0.251 g, 0.00205 mol), was added and the mixture was stirred at room temperature for 48 hours. The progress of the reaction was monitored by TLC (SM was consumed). Water (20 mL) was added to the reaction mixture and extracted with DCM (3 × 50 mL). The resulting organic layer was dried over Na2SO4 and concentrated under reduced pressure to obtain the crude product, which was purified by flash column chromatography (0-100% ethyl acetate in n-hexane) to obtain the pure product tris(4-(heptanoyloxy)butyl)2-((3-(dimethylamino)propanoyl)oxy)propane-1,2,3-tricarboxylate [JC-TL1-12D-E4-6] (compound (12)) (0.3 g, yield 18%) as a brown liquid. 1 H NMR(400MHz,CDCl3):δ 4.18(t,J=6.0Hz,2H),4.10-4.08(m,10H),3.30(d,J=16.0Hz,2H),3.23(d,J=15.6Hz,2H),2.57(t,J=7.2Hz,2H),2.48(t,J=7. 2Hz,2H),2.29(t,J=7.2Hz,6H),2.21(s,6H),1.72-1.67(m,12H),1.64-1.59(m,7H),1.34-1.25(m,17H),0.88(t,J=6.4Hz,9H).

[0671] ELSD analysis: Purity 99.83%, calculated value C 44 H 77 NO 14 =843.53, measured value =844.65 (m / z, M+H + ).

[0672] Example 3. Synthesis protocol for M1266-J03983-027 (JC-TL1-12D-E6-4) (Compound (40)) Synthesis scheme C [ka] Intermediate [C-3]: [ka] A stirred solution of hexane-1,6-diol[C-1] (9.8 g, 0.08 mol) and triethylamine (41.8 g, 0.4 mol) in dichloromethane (150 mL) was cooled to 0°C. 4-(dimethylamino)pyridine-1-ium (1.0 g, 0.008 mol), followed by pentanoyl chloride[C-2] (5.0 g, 0.04 mol), was added, and the reaction mixture was stirred at room temperature for 16 hours. The progress of the reaction was monitored by TLC (SM was consumed). The reaction mixture was quenched to pH 7 with cold saturated NaHCO3 and extracted with DCM (3 × 200 mL). The resulting organic layer was dried over Na2SO4, concentrated under reduced pressure, and the crude product was purified by flash column chromatography (SiO2: 0-20% ethyl acetate in hexane) to obtain the desired 6-hydroxyhexylpentanoate[C-3] (6.2 g, yield 36%) as a pale yellow oil. 1 H NMR(400MHz,CDCl3):δ 4.06(t,J=6.8Hz,2H),3.64(t,J=6.8Hz,2H),2.29(t,J=7.2Hz,2H),1.66-1.54(m,6H),1.42-1.31(m,6H),0.92-0.82(m,3H).

[0673] Intermediate [C-5]: [ka] A stirred solution of 6-hydroxyhexylpentanoate [C-3] (5.3 g, 0.026 mol) and citrate [C-4] (1.26 g, 0.0065 mol) in dichloromethane (100 mL) was cooled to 0°C, and EDC.HCl (5.02 g, 0.026 mol), followed by 4-(dimethylamino)pyridine-1-ium (0.8 g, 0.0065 mol), was added and the mixture was stirred at room temperature for 48 hours. The progress of the reaction was monitored by TLC (SM was consumed). Water (100 mL) was added to the reaction mixture and extracted with DCM (3 × 200 mL). The resulting organic layer was dried over Na2SO4 and concentrated under reduced pressure to obtain the crude product, which was purified by flash column chromatography (0-30% ethyl acetate in n-hexane) to obtain the pure product tris(6-(pentanoyloxy)hexyl)2-hydroxypropane-1,2,3-tricarboxylate[C-5] (1.61 g, yield 33%) as a pale yellow liquid.

[0674] ELSD analysis: Purity 99.62%, calculated value C 39 H 68 O 13 =744.46, measured value =767.20 (m / z, M+Na + ).

[0675] JC-TL1-12D-E6-4 (compound (40)) [ka] A stirred solution of tris(6-(pentanoyloxy)hexyl)2-hydroxypropane-1,2,3-tricarboxylate[C-5] (1.61 g, 0.0022 mol) and 3-(dimethylamino)propanoic acid[C-6] (1.013 g, 0.0086 mol) in dichloromethane (30 mL) was cooled to 0°C, and EDC.HCl (1.69 g, 0.0086 mol), followed by 4-(dimethylamino)pyridine-1-ium (0.26 g, 0.0022 mol), was added and the mixture was stirred at room temperature for 48 hours. The progress of the reaction was monitored by TLC (SM was consumed). Water (20 mL) was added to the reaction mixture and extracted with DCM (3 × 50 mL). The obtained organic layer was dried over Na2SO4 and concentrated under reduced pressure to obtain the crude product, which was purified by flash column chromatography (0-100% ethyl acetate in n-hexane) to obtain the pure product JC-TL1-12D-E6-4 (compound (40)) (0.225 g, yield 12%) as a pale yellow liquid. 1 H NMR(400MHz,CDCl3):δ 4.14(t,J=6.8Hz,2H),4.08-4.03(m,10H),3.30(d,J=15.6Hz,2H),3.21(d,J=15.6Hz,2H),2.60-2.44(m ,4H),2.29(t,J=7.6Hz,6H),2.21(s,6H),1.63-1.56(m,18H),1.37-1.29(m,18H),0.91(t,J=7.2Hz,9H).

[0676] ELSD analysis: Purity 98.58%, calculated value C 44 H 77 NO 14 =843.53, measured value =844.25 (m / z, M+Na + ).

[0677] Example 4. Synthesis protocol for M1266-J03983-025 (JC-TL1-16D-E5-9) (Compound (29)) Synthesis scheme D [ka] Intermediate [D-3]: [ka] A stirred solution of pentane-1,5-diol [D-1] (10 g, 0.0961 mol) and decanoic acid [D-2] (16.5 g, 0.0961 mol) in dichloromethane (200 mL) was cooled to 0°C, and EDC.HCl (22.0 g, 0.115 mol), followed by 4-(dimethylamino)pyridine-1-ium (1.17 g, 0.009 mol), was added and the mixture was stirred at room temperature for 16 hours. The progress of the reaction was monitored by TLC (SM was consumed). Water (100 mL) was added to the reaction mixture and extracted with DCM (3 × 300 mL). The resulting organic layer was dried over Na2SO4 and concentrated under reduced pressure to obtain the crude product, which was purified by flash column chromatography (0-20% ethyl acetate in n-hexane) to obtain the pure product 5-hydroxypentyldecanoate [D-3] (5.4 g, yield 22%) as a pale yellow liquid. 1 H NMR(400MHz,CDCl3):δ 4.07(t,J=6.4Hz,2H),3.68-3.63(m,2H),2.28(t,J=7.6Hz,2H),1.68-1.58(m,6H),1.47-1.42(m,2H),1.29-1.25(m,12H),0.89-0.87(m,3H).

[0678] Intermediate [5]: [ka] A stirred solution of 5-hydroxypentyldecanoate [D-3] (5.37 g, 0.0208 mol) and citric acid [D-4] (1.0 g, 0.0052 mol) in dichloromethane (100 mL) was cooled to 0°C, and EDC.HCl (4.0 g, 0.0208 mol), followed by 4-(dimethylamino)pyridine-1-ium (0.635 g, 0.0052 mol), was added and the mixture was stirred at room temperature for 48 hours. The progress of the reaction was monitored by TLC (SM was consumed). Water (100 mL) was added to the reaction mixture and extracted with DCM (3 × 200 mL). The resulting organic layer was dried over Na2SO4 and concentrated under reduced pressure to obtain the crude product, which was purified by flash column chromatography (0-30% ethyl acetate in n-hexane) to obtain the pure product tris(5-(decanoyloxy)pentyl)2-hydroxypropane-1,2,3-tricarboxylate[D-5] (0.7 g, yield 14%) as a pale yellow liquid. 1 H NMR(400MHz,CDCl3):δ 4.22(t,J=6.8Hz,2H),4.11-4.04(m,10H),2.88(d,J=15.6Hz,2H),2.79(d,J=15.6Hz,2H),2.28 (t,J=7.6Hz,6H),1.74-1.59(m,16H),1.46-1.36(m,6H),1.29-1.26(m,38H),0.89-0.86(m,9H).

[0679] JC-TL1-16D-E5-9 (compound (29)) [ka] A stirred solution of tris(5-(decanoyloxy)pentyl)2-hydroxypropane-1,2,3-tricarboxylate[D-5] (0.7 g, 0.00078 mol) and 3-(dimethylamino)propanoic acid[D-6] (0.36 g, 0.00312 mol) in dichloromethane (20 mL) was cooled to 0°C, and EDC.HCl (0.59 g, 0.00312 mol), followed by 4-(dimethylamino)pyridine-1-ium (0.09 g, 0.00078 mol), was added and the mixture was stirred at room temperature for 48 hours. The progress of the reaction was monitored by TLC (SM was consumed). Water (10 mL) was added to the reaction mixture and extracted with DCM (3 × 25 mL). The resulting organic layer was dried over Na2SO4 and concentrated under reduced pressure to obtain the crude product, which was purified by flash column chromatography (0-100% ethyl acetate in n-hexane) to obtain the pure product JC-TL1-16D-E5-9 (compound (29) (0.11 g, yield 14%)) as a colorless liquid. 1 H NMR(400MHz,CDCl3):δ 4.15(t,J=6.8Hz,2H),4.09-4.04(m,10H),3.30(d,J=15.6Hz,2H),3.22(d,J=15.6Hz,2H),2.60-2.56(m,2H),2.50-2.46(m ,2H),2.28(t,J=7.6Hz,6H),2.22(s,6H),1.69-1.59(m,18H),1.44-1.37(m,6H),1.29-1.26(m,36H),0.87(t,J=6.4Hz,9H).

[0680] ELSD analysis: Purity 98.73%, calculated value C 56 H 101 NO 14 =1011.72, measured value =1012.40 (m / z, M+H + ).

[0681] Example 5. Synthesis protocol for M1266-J03984-031 (TL1-10D-E5-3) (Compound (38)) Synthesis scheme E [ka] Intermediate [E-3]: [ka] A stirred solution of pentane-1,5-diol [E-1] (10 g, 0.096 mol) and triethylamine (48.5 g, 0.48 mol) in dichloromethane (100 mL) was cooled to 0°C. 4-(dimethylamino)pyridine-1-ium (1.17 g, 0.0096 mol), followed by butyryl chloride [E-2] (5.09 g, 0.048 mol), was added, and the reaction mixture was stirred at room temperature for 16 hours. The progress of the reaction was monitored by TLC (SM was consumed). The reaction mixture was quenched to pH 7 with cold saturated NaHCO3 and extracted with DCM (3 × 200 mL). The resulting organic layer was dried over Na2SO4, concentrated under reduced pressure, and the crude product was purified by flash column chromatography (SiO2: 0-20% ethyl acetate in hexane) to obtain the desired 5-hydroxypentyl butyrate [E-3] (4.2 g, yield 25%) as a colorless oil. 1 H NMR(400MHz,CDCl3):δ 4.08(t,J=6.8Hz,2H),3.68-3.63(m,2H),2.28(t,J=7.2Hz,2H),1.70-1.5 8(m,5H),1.47-1.40(m,2H),1.25(t,J=5.2Hz,1H),0.94(t,J=7.6Hz,3H).

[0682] Intermediate [E-5]: [ka] A stirred solution of 5-hydroxypentyl butyrate [E-3] (4.16 g, 0.0239 mol) and citric acid [4] (1.15 g, 0.0059 mol) in dichloromethane (50 mL) was cooled to 0°C, and EDC.HCl (4.60 g, 0.0239 mol), followed by 4-(dimethylamino)pyridine-1-ium (0.73 g, 0.0059 mol), was added and the mixture was stirred at room temperature for 48 hours. The progress of the reaction was monitored by TLC (SM was consumed). Water (100 mL) was added to the reaction mixture and extracted with DCM (3 × 200 mL). The resulting organic layer was dried over Na2SO4 and concentrated under reduced pressure to obtain the crude product, which was purified by flash column chromatography (0-30% ethyl acetate in n-hexane) to obtain the pure product tris(5-(butyryloxy)pentyl)2-hydroxypropane-1,2,3-tricarboxylate[E-5] (1.05 g, yield 27%) as a pale yellow liquid.

[0683] ELSD analysis: Purity 97.05%, calculated value C 33 H 56 O 13 =660.37, measured value =661.10 (m / z, M+H + ).

[0684] JC-TL1-10D-E5-3 (compound (38)) [ka] A stirred solution of tris(5-(butyryloxy)pentyl)2-hydroxypropane-1,2,3-tricarboxylate[E-5] (1.0 g, 0.0015 mol) and 3-(dimethylamino)propanoic acid[E-6] (0.929 g, 0.006 mol) in dichloromethane (20 mL) was cooled to 0°C, and EDC.HCl (1.16 g, 0.006 mol), followed by 4-(dimethylamino)pyridine-1-ium (0.184 g, 0.0015 mol), was added and the mixture was stirred at room temperature for 48 hours. The progress of the reaction was monitored by TLC (SM was consumed). Water (20 mL) was added to the reaction mixture and extracted with DCM (3 × 50 mL). The resulting organic layer was dried over Na2SO4 and concentrated under reduced pressure to obtain the crude product, which was purified by flash column chromatography (0-100% ethyl acetate in n-hexane) to obtain the pure product JC-TL1-10D-E5-3 (compound (38)) (0.410 g, yield 35%) as a pale yellow liquid. 1 H NMR(400MHz,CDCl3):δ 4.15(t,J=6.8Hz,2H),4.12-4.03(m,10H),3.29(d,J=15.6Hz,2H),3.21(d,J=15.6Hz,2H),2.59-2.56(m,2H),2.4 9-2.44(m,2H),2.27(t,J=7.6Hz,6H),2.21(s,6H),1.71-1.60(m,18H),1.44-1.37(m,6H),0.94(t,J=7.6Hz,9H).

[0685] ELSD analysis: Purity 99.79%, calculated value C 38 H 65 NO 14 =759.44, measured value =760.15 (m / z, M+H + ).

[0686] Example 6. Synthesis protocol for M1266-J03996-029 (TL1-14D-E8-4) (Compound (28)) Synthesis scheme F [ka] Intermediate F-3 [ka] A stirred solution of 1,8-octanediol (10 g, 0.084 mol) and triethylamine (45 ml, 0.34 mol) in dichloromethane (200 ml) was cooled to 0°C, and then pentanoyl chloride (4.09 g, 0.034 mol) was added dropwise. The reaction mixture was brought to room temperature and stirred for 16 hours. The progress of the reaction was monitored by ELSD / TLC (SM was consumed). The reaction mixture was washed with DM water (100 mL) and dried over sodium sulfate. The solvent was distilled under reduced pressure, and the resulting crude product was purified with silica using 10% ethyl acetate in n-heptane to obtain the desired product [F-3] (5.1 g, 32.23%) as a yellow liquid. 1H NMR (400MHz, CDCl3): δ 4.04(t,J=6.4Hz,2H),3.63(t,J=6.4Hz,2H),2.29(t,J=7.6Hz,2H),1.61(m,5H),1.36(m,12H),0.90(t,J=6.4Hz,3H).

[0687] Intermediate F-5 [ka] To a stirred solution of 8-hydroxyoctylpentanoate [Int-3] (4.5 g, 0.195 mol) and citric acid (0.937 g, 0.00488 mol) in dichloromethane (20 ml), EDCI (3.75 g, 0.0195 mol) was added, followed by DMAP (0.60 g, 0.00488 mol). The reaction mixture was stirred for 16 hours. The progress of the reaction was monitored by ELSD / TLC (SM was consumed). The reaction mixture was diluted with DCM (25.0 mL), washed with DM water (50 ml), and dried over sodium sulfate. The solvent was distilled, and the resulting crude product was purified with silica using 20% ​​ethyl acetate in n-heptane to obtain the desired product [F-5] (0.8 g, 19.80%) as a yellow liquid.

[0688] LCMS analysis: 99% purity, calculated value C 45 H80 O 13 =828.56, measured value =829.25 (m / z, M+H + ).

[0689] TL1-14D-E8-4 (compound (28)) [ka] To a stirred solution of tris(8-(pentanoyloxy)octyl)2-hydroxypropane-1,2,3-tricarboxylate[F-5] (0.8 g, 0.00964 mol) and N,N-dimethylpropionic acid (0.451 g, 0.00385 mol) in dichloromethane (10 ml), EDCI (0.740 g, 0.00385 mol) was added, followed by DMAP (0.117 g, 0.000964 mol). The reaction mixture was stirred at room temperature for 16 hours. The progress of the reaction was monitored by ELSD / TLC (SM was consumed). The reaction mixture was diluted with DCM (25.0 mL), washed with DM water (50 ml), and dried over sodium sulfate. The solvent was distilled, and the resulting crude product was purified with silica using 10% ethyl acetate in n-heptane to obtain the desired product TL1-14D-E8-4 (0.30 g, 25%) as a yellow liquid. 1H NMR(400MHz,DMSO-d6):δ 4.13(t,J=6.4Hz,2H),4.06(m,10H),3.31-3.19(m,4H),2.58(t,J=7.2Hz,2H)2.48(t,J=7.2Hz,2H )2.29(t,J=7.6Hz,6H),2.21(s,6H),1.64-1.56(m,17H),1.38-1.31(m,31H),0.91(t,J=7.2,9H).

[0690] LCMS analysis: purity 99.34%, calculated value C 50 H 89 NO 14 =927.63, measured value =928.25 (m / z, M+H + ).

[0691] Example 7. Synthesis protocol for M1266-J03996-019 (TL1-16D-E6-8) (Compound (57)) Synthesis scheme G [ka] Intermediate G-3 [ka] A stirred solution of 1,6-hexanediol (10 g, 0.084 mol) and trimethylamine (55.92 ml, 0.42 mol) in dichloromethane (200 ml) was cooled to 0°C. Nonanoyl chloride (7.42 g, 0.042 mol) was added dropwise. The reaction mixture was stirred at room temperature for 16 hours. The progress of the reaction was monitored by ELSD / TLC (SM was consumed). The reaction mixture was washed with DM water (100.0 mL), dried over sodium sulfate, and concentrated. The resulting crude product was purified with silica using 10% ethyl acetate in n-heptane to obtain the desired product [G-3] (5.0 g, 22.90%) as a yellow liquid. 1H NMR (400MHz, CDCl3): δ 4.06(t,J=6.4Hz,2H),3.64(m,2H),2.28(t,J=7.6Hz,2H),1.66-1.59(m,6H),1.43-1.20(m,14H),0.89(t,J=6.4Hz,3H).

[0692] Intermediate G-5 [ka] To a stirred solution of 6-hydroxyhexyl nonanoate [G-3] (4.5 g, 0.0174 mol) and citric acid (1.06 g, 0.0055 mol) in dichloromethane (100 ml), EDCI (4.25 g, 0.022 mol), followed by DMAP (0.076 g, 0.0055 mol), was added. The reaction mixture was stirred for 16 hours. The progress of the reaction was monitored by ELSD / TLC (SM was consumed). The reaction mixture was washed with DM water (100 mL), dried over sodium sulfate, and concentrated. The resulting crude product was purified with silica using 10% ethyl acetate in n-heptane to obtain the desired product [G-5] 0.7 g (13.39%) as a yellow liquid mass.

[0693] LCMS analysis: purity 90.81%, calculated value C 51 H 92 O 13 =912.65, measured value =913.35 (m / z, M+H+).

[0694] TL1-16D-E6-8 (compound (57)) [ka] To a stirred solution of tris(6-(nonanoyloxy)hexyl)2-hydroxypropane-1,2,3-tricarboxylate[G-5] (0.7 g, 0.000766 mol) and N,N-dimethylpropionic acid[6] (0.358 g, 0.00306 mol) in dichloromethane (25 ml), EDCI (0.590 g, 0.003061 mol) was added, followed by DMAP (0.11 g, 0.00090 mol). The reaction mixture was stirred at room temperature for 16 hours. The progress of the reaction was monitored by ELSD / TLC (SM was consumed). The reaction mixture was quenched with DM water (100 ml), extracted with DCM (2 × 50.0 mL), dried over sodium sulfate, and concentrated. The crude product was purified with silica using 10% ethyl acetate in n-heptane to obtain the desired product [TL1-16D-E6-8] 0.250 g (32.25%) as a yellow liquid mass. 1H NMR(400MHz,DMSO-d6):δ 4.21 -4.15(t,J=6.4Hz,2H),4.08(m,10H),3.31-3.19(m,4H),2.58(t,J=7.2Hz,2H)2.48(m,2H)2.29(t, J=7.6Hz,6H),2.218(s,6H),1.61(br,18H),1.37(bs,12H),1.28-1.26(m,30H)0.88(t,J=7.2,9H).

[0695] LCMS analysis: purity 96.13%, calculated value C 56 H 101 NO 14 =1011.72, measured value =1012.35 (m / z, M+H + ).

[0696] Example 8. Synthesis protocol for M1266-J03996-031 (TL1-16D-E8-6) (Compound (54)) Synthesis scheme H [ka] Intermediate H-3 [ka] A stirred solution of 1,8-octanediol (10 g, 0.084 mol) and triethylamine (45 ml, 0.34 mol) in dichloromethane (200 ml) was cooled to 0°C. Heptanoyl chloride (5.05 g, 0.034 mol) was added dropwise. The reaction mixture was brought to room temperature and stirred for 16 hours. The progress of the reaction was monitored by ELSD / TLC (SM was consumed). The reaction mixture was washed with 0.1 L of DM water, the dichloromethane layer was separated and dried over sodium sulfate. The solvent was distilled under reduced pressure, and the resulting crude product was purified with silica using 10% ethyl acetate in n-heptane to obtain 5.2 g (29.42%) of [H-3] as a yellow liquid mass. 1H NMR (400MHz, CDCl3): δ 4.04(t,J=6.4Hz,2H),3.62(t,J=6.4Hz,2H),2.28(t,J=7.6Hz,2H),1.62(m,6H),1.38(m,15H),0.87(t,J=6.4Hz,3H).

[0697] Intermediate H-5 [ka] To a stirred solution of 8-hydroxyoctylheptanoate [H-3] (5.0 g, 0.143 mol) and citric acid (0.928 g, 0.00483 mol) in dichloromethane (100 ml), EDCI (3.71 g, 0.0193 mol) was added, followed by DMAP (0.58 g, 0.0193 mol). The reaction mixture was stirred at room temperature for 16 hours. The progress of the reaction was monitored by ELSD / TLC (SM was consumed). The reaction mixture was washed with DM water (50 ml) and dried over sodium sulfate. The solvent was distilled, and the resulting crude product was purified with silica using 10% ethyl acetate in n-heptane to obtain the desired product [Int-5] (0.820 g, 18.63%) as a yellow liquid mass.

[0698] LCMS analysis: purity 98.36%, calculated value C 51 H 92 O 13 =912.65, measured value =913.35 (m / z, M+H + ).

[0699] TL1-16D-E8-6 (compound (54)) [ka] To a stirred solution of tris(8-(heptanoyloxy)octyl)2-hydroxypropane-1,2,3-tricarboxylate[H-5] (0.820 g, 0.00897 mol) and N,N-dimethylpropionic acid[Int-6] (0.420 g, 0.00359 mol) in dichloromethane (50 ml), EDCI (0.690 g, 0.00359 mol) was added, followed by DMAP (0.109 g, 0.000897 mol). The reaction mixture was stirred at room temperature for 16 hours. The progress of the reaction was monitored by ELSD / TLC (SM was consumed). The reaction mixture was washed with DM water (50 ml) and dried over sodium sulfate. The solvent was distilled, and the resulting crude product was purified with silica using 20% ​​ethyl acetate in n-heptane to obtain the desired product [TL1-16D-E8-6] 0.250 g (27.77%) as a yellow liquid mass. 1H NMR(400MHz,DMSO-d6):δ 4.127(t,J=6.4Hz,2H),4.061(m,10H),3.310-3.190(m,4H),2.583(t,J=7.2Hz,2H)2.478(t,J=7.2Hz,2H )2.84(t,J=7.6Hz,6H),2.215(s,6H),1.699(s,1H),1.607(m,17H),1.311(m,44H),0.877(t,J=7.2,9H).

[0700] LCMS analysis: purity 98.95%, calculated value C 56 H 101 NO 14 =1011.72, measured value =1012.30 (m / z, M+H + ).

[0701] Example 9. Synthesis protocol for M1266-J03996-021 (TL1-18D-E6-10) (Compound (56)) Synthesis Scheme I [ka] Intermediate I-3 [ka] A stirred solution of 1,6-hexanediol (10 g, 0.084 mol) and triethylamine (55.92 ml, 0.42 mol) in dichloromethane (200 ml) was cooled to 0°C, and undecanoyl chloride (8.59 g, 0.042 mol) was added dropwise. The reaction mixture was stirred at room temperature for 16 hours. The progress of the reaction was monitored by ELSD / TLC (SM was consumed). The reaction mixture was washed with DM water (100 mL) and dried over sodium sulfate. The solvent was distilled under reduced pressure, and the resulting crude product was purified with silica using 10% ethyl acetate in n-heptane. 7.3 g by weight of pure I-3 (30.12%) was obtained as a yellow liquid mass. 1H NMR (400MHz, CDCl3):δ 4.06(t,J=6.4Hz,2H),3.64(t,J=6.4Hz,2H),2.28(t,J=7.6Hz,2H),1.63(m,5H),1.43(m,4H),1.25(m,15H),0.87(t,J=6.4Hz,3H).

[0702] Intermediate I-5 [ka] To a stirred solution of 6-hydroxyhexyl undecanoate [I-3] (7.0 g, 0.02443 mol) and citric acid [4] (1.48 g, 0.0077 mol) in dichloromethane (100 ml), EDCI (5.95 g, 0.0310 mol) was added, followed by DMAP (0.946 g, 0.0077 mol). The reaction mixture was stirred at room temperature for 16 hours. The progress of the reaction was monitored by ELSD / TLC (SM was consumed). The reaction mixture was washed with DM water (500 ml), dried over sodium sulfate, and concentrated. The crude product was purified with silica using 10-15% ethyl acetate in n-heptane to obtain the desired product [I-5] 0.550 g (7.25%) as a yellow liquid mass.

[0703] LCMS analysis: purity 99.54%, calculated value C 57 H 104 O 13 =996.75, measured value = 1019.40 (m / z, M+H ++23).

[0704] TL1-18D-E6-10 (compound (56)) [ka] To a stirred solution of tris(6-(undecanoyloxy)hexyl)2-hydroxypropane-1,2,3-tricarboxylate[I-5] (0.550 g, 0.00055 mol) and N,N-dimethylpropionic acid[I-6] (0.258 g, 0.002207 mol) in dichloromethane (25 ml), EDCI (0.423 g, 0.002207 mol) was added, followed by DMAP (0.07 g, 0.00055 mol). The reaction mixture was stirred at room temperature for 16 hours. The progress of the reaction was monitored by ELSD / TLC (SM was consumed). The reaction mixture was washed with DM water (25 ml), dried over sodium sulfate, and concentrated. The crude product was purified with silica using 10% ethyl acetate in n-heptane. The resulting pure product [TL1-18D-E6-10] was obtained as a yellow liquid in the form of 0.150 g (24.83%). 1H NMR (400MHz, CDCl3):δ 4.14(t,J=6.4Hz,2H),4.08(m,10H),3.31-3.19(m,4H),2.59(t,J=7.2Hz,2H)2.48(t,J=7.2Hz,2H) 2.28(t,J=7.6Hz,6H),2.22(s,6H),1.61(m,18H),1.37(br,12H),1.28(m,42H)0.87(t,J=7.2,9H).

[0705] LCMS analysis: purity 99.87%, calculated value C 62 H 113 NO 14 =1095.82, measured value =1096.40 (m / z, M+H + ).

[0706] Example 10. Synthesis protocol for TL1-18D-E8-8 (compound (53)) Synthesis scheme J [ka] Intermediate J-3 [ka] A stirred solution of octanediol (10 g, 0.068 mol) in dichloromethane (200 ml) was cooled to 0°C, and triethylamine (45.26 ml, 0.34 mol) was added. The reaction mixture was stirred for 0.5 minutes, and then DMAP (4.17 g, 0.034 mol) and then nonanoyl chloride (6.0 g, 0.034 mol) were added dropwise. The resulting reaction mixture was stirred at room temperature for 16 hours. The progress of the reaction was monitored by ELSD / TLC (SM was consumed). The reaction mixture was washed with DM water (100 mL), dried over sodium sulfate, and concentrated. The resulting crude product was purified with silica using 10% ethyl acetate in n-heptane to obtain [J-3] (5.2 g, 26.55%) as a yellow liquid mass. 1H NMR (400MHz, CDCl3): δ 4.04(t,J=6.4Hz,2H),3.65(m,2H),2.28(t,J=7.6Hz,2H),1.57(m,4H),1.280(m,20H),0.888(t,J=6.4Hz,3H).

[0707] Intermediate J-5 [ka] To a stirred solution of 8-hydroxyoctyl nonanoate [J-3] (5.0 g, 0.0174 mol) and citric acid (1.06 g, 0.0055 mol) in dichloromethane (100 ml), EDCI (4.25 g, 0.022 mol) was added, followed by DMAP (0.076 g, 0.0055 mol). The reaction mixture was stirred at room temperature for 16 hours. The progress of the reaction was monitored by ELSD / TLC (SM was consumed). The reaction mixture was diluted with DCM (100.0 mL), washed with DM water (100.0 mL), dried over sodium sulfate, and concentrated. The resulting crude product was purified with silica using 10% ethyl acetate in n-heptane to obtain [I-5] (0.9 g, 15.7%) as a yellow liquid mass. 1H NMR(400MHz,DMSO-d6):δ 6.92(s,1H),4.20-4.15(m,4H),4.11(m,8H),4.01(s,2H),3.95(s,2H),2.28(t,J=7.6Hz,6H),1.63(m,18H),1.33(m,52H)0.83(m,9H).

[0708] TL1-18D-E8-8 (compound (53)) [ka] To a stirred solution of tris(8-(nonanoyloxy)octyl)2-hydroxypropane-1,2,3-tricarboxylate[I-5] (0.9 g, 0.00090 mol) and N,N-dimethylpropionic acid (0.422 g, 0.0036 mol) in dichloromethane (10 ml), EDCI (0.7 g, 0.00361 mol) was added, followed by DMAP (0.11 g, 0.00090 mol). The reaction mixture was stirred at room temperature for 16 hours. The progress of the reaction was monitored by ELSD / TLC (SM was consumed). The reaction mixture was diluted with (50.0 mL), washed with DM water (100 ml), dried on sodium sulfate, and concentrated. The crude product obtained was purified with silica using 10% ethyl acetate in n-heptane to obtain the final [TL1-18D-E8-8] (0.288 g, 29.12%) as a yellow liquid mass. 1 H NMR(400MHz,CDCl3):δ 4.11(t,J=6.8Hz,2H),4.01(m,10H),3.31-3.27(d,J=15.6Hz,2H),3.23-3.19(d,J=15.6Hz,2H),2.60-2.56(m,2H),2.49-2.4 6(m,2H),2.28(t,J=7.6Hz.6H),2.21(s,6H),1.66-1.57(m,18H),1.31-1.26(m,54H),1.24(brs,28H),0.87(t,J=6.8Hz,9H).

[0709] LCMS analysis: purity 99.37%, calculated value C 62 H 113 NO 14=1095.82, measured value =1096.40 (m / z, M+H + ).

[0710] Example 11. Synthesis protocol for M1266-J03983-035 (JC-TL1-18D-E4-12) (Compound (22)) Synthesis scheme K [ka] Intermediate [K-3]: [ka] A stirred solution of butane-1,4-diol [K-1] (10 g, 0.111 mol) and tridecanoic acid [K-2] (23.7 g, 0.111 mol) in dichloromethane (300 mL) was cooled to 0°C, and EDC.HCl (25.5 g, 0.133 mol), followed by 4-(dimethylamino)pyridine-1-ium (6.7 g, 0.055 mol), was added and the mixture was stirred at room temperature for 16 hours. The progress of the reaction was monitored by TLC (SM was consumed). Water (100 mL) was added to the reaction mixture and extracted with DCM (3 × 300 mL). The resulting organic layer was dried over Na₂SO₄ and concentrated under reduced pressure to obtain the crude product, which was purified by flash column chromatography (0-20% ethyl acetate in n-hexane) to obtain the pure product 4-hydroxybutyl tridecanoate [K-3] (11.8 g, yield 35%) as a pale yellow liquid. 1 H NMR (400MHz, CDCl3): δ 4.10(t,J=6.4Hz,2H),3.71-3.66(m,2H),2.28(t,J=7.6Hz,2H),1.76-1.57(m,6H),1.27-1.11(m,18H),0.89-0.85(m,3H).

[0711] Intermediate [K-5]: [ka] A stirred solution of 4-hydroxybutyl tridecanoate [K-3] (11.8 g, 0.04123 mol) and citric acid [K-4] (2.0 g, 0.0103 mol) in dichloromethane (150 mL) was cooled to 0°C, and EDC.HCl (7.90 g, 0.0412 mol), followed by 4-(dimethylamino)pyridine-1-ium (1.25 g, 0.0103 mol), was added and the mixture was stirred at room temperature for 48 hours. The progress of the reaction was monitored by TLC (SM was consumed). Water (100 mL) was added to the reaction mixture and extracted with DCM (3 × 200 mL). The resulting organic layer was dried over Na2SO4 and concentrated under reduced pressure to obtain the crude product, which was purified by flash column chromatography (0-30% ethyl acetate in n-hexane) to obtain the pure product tris(4-(tridecanoyloxy)butyl)2-hydroxypropane-1,2,3-tricarboxylate[K-5] (1.1 g, yield 10%) as a pale yellow liquid.

[0712] ELSD analysis: Purity 99.74%, calculated value C 57 H 104 O 13 =996.75, measured value = 1019.30 (m / z, M+Na + ).

[0713] JC-TL1-18D-E4-12 (compound (22)) [ka] A stirred solution of tris(4-(tridecanoyloxy)butyl)2-hydroxypropane-1,2,3-tricarboxylate [K-5] (1.1 g, 0.0011 mol) and 3-(dimethylamino)propanoic acid [K-6] (0.51 g, 0.0044 mol) in dichloromethane (20 mL) was cooled to 0°C, and EDC.HCl (0.84 g, 0.0044 mol), followed by 4-(dimethylamino)pyridine-1-ium (0.134 g, 0.0011 mol), was added and the mixture was stirred at room temperature for 48 hours. The progress of the reaction was monitored by TLC (SM was consumed). Water (10 mL) was added to the reaction mixture and extracted with DCM (3 × 25 mL). The obtained organic layer was dried over Na2SO4 and concentrated under reduced pressure to obtain the crude product, which was purified by flash column chromatography (0-100% ethyl acetate in n-hexane) to obtain the pure product tris(4-(tridecanoyloxy)butyl)2-((3-(dimethylamino)propanoyl)oxy)propane-1,2,3-tricarboxylate [JC-TL1-18D-E4-12] (compound (22)) (0.2 g, yield 16%) as a colorless liquid. 1 H NMR(400MHz,CDCl3):δ 4.19-4.16(m,2H),4.10-4.08(m,10H),3.30(d,J=15.6Hz,2H),3.23(d,J=15.6Hz,2H),2.59-2.56(m,2H),2.49-2.46(m,2 H),2.28(t,J=7.6Hz,6H),2.21(s,6H),1.70-1.69(m,12H),1.62-1.59(m,7H),1.31-1.28(m,53H),0.87(t,J=6.8Hz,9H).

[0714] ELSD analysis: Purity 99.91%, calculated value C 62 H 113 NO 14 =1095.82, measured value =1096.45 (m / z, M+H + ).

[0715] Example 12. Synthesis protocol for M1266-J03996-027 (TL1-12D-E8-2) (Compound (11)) Synthesis scheme L [ka] Intermediate L-3 [ka] A stirred solution of 1,8-octanediol (10 g, 0.084 mol) and triethylamine (45 ml, 0.34 mol) in dichloromethane (200 ml) was cooled to 0°C, and propionyl chloride (3.14 g, 0.034 mol) was added dropwise. The reaction mixture was stirred at room temperature for 16 hours. The progress of the reaction was monitored by ELSD / TLC (SM was consumed). The reaction mixture was washed with DM water (100.0 mL), dried over sodium sulfate, and concentrated under reduced pressure. The crude product was purified with silica using 10% ethyl acetate in n-heptane to obtain the desired product [L-3] (7.3 g; 30.12%) as a yellow liquid. 1H NMR (400MHz, CDCl3): δ 4.05(t,J=6.4Hz,2H),3.62(q,J=6.0Hz,2H),2.30(q,J=7.6Hz,2H),1.61(m,3H),1.35(s,9H),1.11(t,J=6.4Hz,3H).

[0716] Intermediate L-5 [ka] To a stirred solution of 8-hydroxyoctylpropionate [L-3] (5.3 g, 0.062 mol) and citric acid (1.25 g, 0.00655 mol) in dichloromethane (20 ml), EDCI (5.03 g, 0.0262 mol) was added, followed by DMAP (0.80 g, 0.00635 mol). The reaction mixture was stirred at room temperature for 16 hours. The progress of the reaction was monitored by ELSD / TLC (Sm was consumed). The reaction mixture was washed with DM water (50 ml), dried over sodium sulfate, and concentrated. The crude product was purified with silica using 10% ethyl acetate in n-heptane to obtain the desired product [Int-5] (1.1 g, 22.68%) as a yellow liquid. 1H NMR (400MHz, CDCl3): δ 4.059(t,J=6.8Hz,6H),3.643(d,J=3.2Hz,6H),2.282(t,J=7.6Hz,6H),1.629(m,18H),1.331(m,22H)0.829(m,9H).

[0717] LCMS analysis: purity 98.35%, calculated value C 39 H 68 O 13 =744.47, measured value =745.20 (m / z, M+H + ).

[0718] TL1-12D-E8-2 (compound (11)) [ka] To a stirred solution of tris(8-(propionyloxy)octyl)2-hydroxypropane-1,2,3-tricarboxylate[L-5] (1.1 g, 0.00147 mol) and N,N-dimethylpropionic acid (0.691 g, 0.00590 mol) in dichloromethane (10 ml), EDCI (1.13 g, 0.00590 mol) was added, followed by DMAP (0.180 g, 0.00147 mol). The reaction mixture was stirred at room temperature for 16 hours. The progress of the reaction was monitored by ELSD / TLC (SM was consumed). The reaction mixture was washed with 20 ml of DM water and dried over sodium sulfate. The solvent was distilled, and the resulting crude product was purified with silica using 10% ethyl acetate in n-heptane to obtain the desired product [TL1-12D-E8-2](compound (0.300 g, 25%) as a yellow liquid). 1H NMR (400MHz, CDCl3):δ 4.13(t,J=6.4Hz,2H),4.08(t,J=6.8Hz,10H),3.31-3.18(m,4H),2.61(t,J=7.2Hz,2H)2.50(t,J=7. 2Hz,2H)2.34-2.28(q,J=7.6Hz,6H),2.24(s,6H),1.61(m,12H),1.31(brs,24H),1.13(t,J=7.2,9H).

[0719] LCMS analysis: purity 99.77%, calculated value C 44 H 77 NO 14 =843.53, measured value =844.15 (m / z, M+H + ).

[0720] Example 13. Synthesis protocol for M1266-J03996-055 (TL1-14D-E5-7-2i) (Compound (48)) Synthesis scheme M [ka] Intermediate M-3 [ka] A stirred solution of 1,5-pentanediol (20 g, 0.190 mol) and triethylamine (90 ml, 0.68 mol) in dichloromethane (200 ml) was cooled to 0°C. 3-Pentyloctanoyl chloride (22.13 g, 0.095 mol) was added dropwise. The reaction mixture was brought to room temperature and stirred for 16 hours. The progress of the reaction was monitored by ELSD / TLC (SM was consumed). The reaction mixture was washed with DM water (100 mL), dried over sodium sulfate, and concentrated under reduced pressure. The resulting crude product was purified with silica using 10% ethyl acetate in n-heptane to obtain the desired product [M-3] (17.0 g, 60.71%) as a yellow liquid. 1H NMR (400MHz, CDCl3):δ 4.07(t,J=6.4Hz,2H),3.66(t,J=6.4Hz,2H),2.23(d,J=6.8Hz,2H),1.83(s,1 H),1.69-1.58(m,4H),1.47-1.41(m,2H),1.25(s,17H),0.88(t,J=6.4Hz,6H).

[0721] LCMS analysis: purity 99.96%, calculated value C 18 H 36 O3 = 300.27, measured value = 301.30 (m / z, M+H + ).

[0722] Intermediate M-5 [ka] To a stirred solution of 5-hydroxypentyl-3-pentyloctanoate [M-3] (17 g, 0.056 mol) and citric acid (2.72 g, 0.0141 mol) in dichloromethane (60 ml), EDCI (10.82 g, 0.0056 mol), followed by DMAP (1.72 g, 0.0141 mol), was added. The reaction mixture was stirred for 16 hours. The progress of the reaction was monitored by ELSD / TLC (SM was consumed). The reaction mixture was washed with DM water (50 ml) and dried over sodium sulfate. The solvent was distilled, and the resulting crude product was purified with silica using 10% ethyl acetate in n-heptane to obtain the desired product [Int-5] (12 g, 85%) as a yellow liquid mass. 1H NMR (400MHz, CDCl3):δ 4.22(t,J=6.8Hz,2H),4.05(m,10H),2.89-2.77(m,4H),2.23-2.21(d,J=7.2Hz,6H),1.8 3(m,3H),1.72-1.61(m,12H)1.56(m,6H),1.39(m,8H),1.32-1.26(m,40H),0.85(m,18H).

[0723] TL1-14D-E5-7-2i (compound (48)) [ka] To a stirred solution of tris(5-((3-pentyloctanoyl)oxy)pentyl)2-hydroxypropane-1,2,3-tricarboxylate [Int-5] (2.5 g, 0.00240 mol) and N,N-dimethylpropionic acid [Int-6] (1.12 g, 0.00962 mol) in dichloromethane (100 ml), EDCI (2.0 g, 0.00962 mol) was added, followed by DMAP (0.30 g, 0.0024 mol). The reaction mixture was stirred at room temperature for 16 hours. The progress of the reaction was monitored by ELSD / TLC (SM was consumed). The reaction mixture was washed with DM water (50 ml) and dried over sodium sulfate. The solvent was distilled, and the resulting crude product was purified with silica using 30% ethyl acetate in n-heptane to obtain 1.2 g (44.44%) of the desired product [TL1-14D-E5-7-2i] as a yellow liquid mass. 1H NMR (400MHz, CDCl3):δ 4.15(t,J=6.4Hz,2H),4.08(m,10H),3.31-3.19(m,4H),2.59(t,J=7.2Hz,2H),2.47(t,J=7.2Hz,2H),2.2 2-2.21(m,12H),1.83(m,3H),1.69-1.60(m,12H),1.42(m,6H),1.30-1.22(m,48H),0.87(t,J=7.2,18H).

[0724] LCMS analysis: purity 99.83%, calculated value C 65 H 119 NO 14 =1137.86, measured value =1138.45 (m / z, M+H + ).

[0725] Example 14. Synthesis protocol for M1266-450487 (JC-ATL-011) (Compound (6)) Synthesis scheme N [ka] Intermediate N-5 [ka] To a solution of citric acid (25.0 g, 0.130 mol) and octan-1-ol (67.7 g, 0.52 mol) in toluene (500 mL), PTSA (13.0 g, 0.065 mol) was added, and the resulting mixture was stirred at 100 °C for 24 hours. The progress of the reaction was monitored by TLC (5% HCl / hexane). The reaction mixture was cooled to room temperature, diluted with water (500 mL), and extracted with ethyl acetate (500 mL x 3). The organic layer was dried over anhydrous Na₂SO₄, and the solvent was evaporated. The crude product was purified by column chromatography using 60-120 mesh silica, and eluted with 5% ethyl acetate in hexane to obtain compound [N-5] (40.0 g, 58.22%) as a colorless liquid. 1 H NMR(400MHz,DMSD-d6):δ 5.61(s,1H),4.01(t,J=6.8Hz,2H),3.96(t,J=6.4Hz,4H),2.87(d,J=15.2Hz,2H), 2.72(d,J=15.2Hz,2H),1.57-1.49(m,6H),1.24(brs,30H),0.85(t,J=6.4Hz,9H).

[0726] LCMS analysis: purity 97.38%, calculated value C 30 H 56 O7 = 528.40, measured value = 529.2 (m / z, M+H+).

[0727] JC-ATL-011 (compound (6)) [ka] To a stirred solution of 1,2,3-trioctyl 2-hydroxypropane-1,2,3-tricarboxylate [N-5] (0.5 g, 946 μmol) in dichloromethane (30 mL, 469 mmol), triethylamine (287 mg, 3 equivalents, 2.84 mmol) and 4-(dimethylamino)pyridine-1-ium (116 mg, 946 μmol) were added at 0°C. The reaction mixture was stirred at the same temperature for 10 minutes, followed by the addition of ditrichloromethyl carbonate (281 mg, 946 μmol) and 2-(dimethylamino)ethane-1-ol (84.3 mg, 946 μmol). The resulting reaction mixture was stirred at room temperature for 16 hours. The progress of the reaction was monitored by TLC. The resulting reaction mixture was quenched with water (50 mL) and extracted with dichloromethane (3 × 30 mL). The entire organic layer was combined, dried over Na2SO4, and concentrated under reduced pressure. The resulting crude material was purified by column chromatography using 0-5% MeOH in DCM as the eluent. The pure fractions were combined and concentrated under reduced pressure to obtain [JC-ATL-011] (compound (6)) (410 mg, yield -67.37%) as a yellow liquid. 1 H NMR(400MHz,CDCl3):δ 4.20(t,J=6.0Hz,2H),4.16-4.14(d,J=6.8Hz,2H),3.35-3.31(d,J=15.6Hz,2H),3.24-3.20(d,J=15. 6Hz,2H),2.58(d,J=6Hz,2H),2.27(s,6H),1.67-1.56(m,6H),1.27(brs,30H),0.87(t,J=6.4Hz,9H).

[0728] LCMS analysis: purity 99.87%, calculated value C 35 H 65 NO9 = 644.47, Measured value = 644.75 (m / z, M+H + ).

[0729] Example 15. Synthesis protocol for M1266-449313 (JC-ATL-012) (Compound (25)) Synthesis scheme O [ka] Intermediate O-3 [ka] To a stirred solution of pentane-1,5-diol (6.5 g, 0.9 equivalents, 62.4 mmol) in dichloromethane (50 mL, 781 mmol), octanoic acid (10 g, 69.3 mmol) was added. The reaction mixture was cooled to 0°C. ({[3-(dimethylamino)propyl]imino}methylidene)(ethyl)amine hydrochloride (16.7 g, 1.3 equivalents, 87.4 mmol) and 4-(dimethylamino)pyridine-1-ium (1.71 g, 0.2 equivalents, 13.9 mmol) were added. The reaction mixture was stirred at room temperature for 48 hours. The progress of the reaction was monitored by TLC / ELSD, and after the starting materials were consumed, the reaction mixture was washed with saturated NaHCO3 and brine solution. After the organic layer was evaporated, the resulting crude product was purified by flash column chromatography to obtain compound O-3 (6.0 g, yield: 37.56%) as a colorless liquid. 1 H NMR(400MHz,CDCl3):δ 4.08(t,J=6.4Hz,2H),3.67(s,2H),2.30(t,J=7.2Hz,2H),1.71-1.58(m,6H),1.44(m,2H),1.29(s,10H),0.89(t,J=6.8Hz,3H).

[0730] Intermediate O-5 [ka] To a stirred solution of 2-hydroxypropane-1,2,3-tricarboxylic acid [compound O-3] (250 mg, 1.3 mmol) in dichloromethane (5 mL, 78.1 mmol), N,N-dimethylpyridine-4-amine (159 mg, 1.3 mmol) and ({[3-(dimethylamino)propyl]imino}methylidene)(ethyl)amine hydrochloride (998 mg, 4 equivalents, 5.2 mmol) were added. The reaction mixture was stirred at 25°C for 10 minutes, and then 5-hydroxypentyl octanoate (1.35 g; 5.86 mmol) was added. The reaction mixture was stirred at room temperature under a nitrogen atmosphere for 16 hours. The progress of the reaction was monitored by TLC / ELSD, and after the starting materials were consumed, the reaction mixture was diluted with water and dichloromethane. The organic layer was separated and dried over sodium sulfate. The solvent was distilled under reduced pressure. The crude product was purified by column chromatography using 15-20% ethyl acetate to obtain compound O-5 (0.9 g; yield: 83.42%) as a pale yellow liquid. 1 H NMR(400MHz,CDCl3):δ 4.21(m,2H),4.08(m,10H),2.90-2.77(dd,J=15.6Hz.4H),2.29(t,J=7.6H z,6H),1.61(m,12H),1.41(m,8H),1.28(brs,28H),0.85(t,J=6.4Hz,9H).

[0731] JC-ATL-012 (compound (25)) [ka] To a stirred solution of 1,2,3-tris[5-(octanoyloxy)pentyl]2-hydroxypropane-1,2,3-tricarboxylate [compound O-5] (0.5 g, 603 μmol) in dichloromethane (6 mL, 93.7 mmol), triethylamine (183 mg, 3 equivalents, 1.81 mmol) and 4-(dimethylamino)pyridine-1-ium (74.3 mg, 603 μmol) were added successively at 0°C, and the mixture was stirred at the same temperature for 10 minutes. Subsequently, ditrichloromethyl carbonate (179 mg, 603 μmol) and 2-(dimethylamino)ethane-1-ol (53.8 mg, 603 μmol) were added. The resulting reaction mixture was stirred at room temperature for 16 hours, and the progress of the reaction was monitored by TLC / ELSD. The resulting reaction mixture was quenched with water (50 mL) and extracted with dichloromethane (3 × 30 mL). The combined organic layers were dried over Na2SO4 and concentrated under reduced pressure. The resulting crude material was purified by column chromatography using 0-5% MeOH in DCM as the eluent. The pure fractions were combined and concentrated under reduced pressure to obtain [JC-ATL-012] (compound (25)) (225 mg, yield 39.51%) as a yellow liquid. 1 H NMR(400MHz,CDCl3):δ 4.195(m,4H),4.08(m,10H),3.34-3.30(d,J=15.6Hz,2H),3.24-3.20(d,J=15.6Hz,2H),2.57(t,J=5.6H z,2H),2.28(t,J=8.8Hz,12H),1.72-1.57(m,19H),1.44(m,6H),1.28(brs,24H),0.87(t,J=6.4Hz,9H).

[0732] LCMS analysis: purity 99.74%, calculated value C 50 H 89 NO 15 =943.62, measured value =944.60 (m / z, M+H + ).

[0733] Example 16. Synthesis protocol for M1266-674154 (TL1-14D-E5-7-6i) (Compound (36)) Synthesis scheme P [ka] Intermediate P-3 [ka] To a stirred solution of 7-methyloctanoic acid (12 g, 75.8 mmol) in dichloromethane (0.2 L), N,N-dimethylpyridine-4-amine (9.26 g, 75.8 mmol) and ({[3-(dimethylamino)propyl]imino}methylidene)(ethyl)amine hydrochloride (29.1 g, 2 equivalents, 152 mmol) were slowly added at 0°C. After 10 minutes, pentane-1,5-diol (15.8 g, 2 equivalents, 152 mmol) was added. The reaction mixture was stirred at room temperature for 18 hours. The progress of the reaction was monitored by ELSD / TLC. The reaction mixture was diluted with cold water (500.0 mL) and extracted with dichloromethane (3 × 100 mL). The combined organic layers were washed with fresh water (2 × 100 mL) and brine solution (2 × 100.0 mL). The organic layers were dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. The crude product was purified by column chromatography using 10% ethyl acetate in hexane to obtain 5-hydroxypentyl 7-methyloctanoic acid [Int-3] (12 g, 49.1 mmol) as a colorless, clear liquid. 1 H NMR(400MHz,CDCl3):δ 4.07(t,J=6.8Hz,2H),3.68-3.63(dd,J=5.2Hz,2H),2.29(t,J=7.6Hz,2H),1.6 4(m,5H),1.58-1.41(m,3H),1.29(m,5H),1.16(m,2H),0.851(d,J=6.8Hz,6H).

[0734] Intermediate P-5 [ka] A solution of 2-hydroxypropane-1,2,3-tricarboxylic acid (2.44 g, 12.7 mmol) in dichloromethane (0.1 L) was cooled to 0°C. Then, N,N-dimethylpyridine-4-amine (1.55 g, 12.7 mmol) and ({[3-(dimethylamino)propyl]imino}methylidene)(ethyl)amine hydrochloride (9.73 g, 4 equivalents, 50.7 mmol) were slowly added at room temperature. After 10 minutes, 5-hydroxypentyl 7-methyloctanoate [P-3] (12.4 g, 4 equivalents, 50.7 mmol) was added dropwise. The reaction mixture was stirred at 25°C for 16 hours. The progress of the reaction was monitored by TLC. The reaction mixture was diluted with cold water (100.0 mL) and extracted with dichloromethane (3 × 100 mL). The combined organic layers were washed with fresh water (2 × 50 mL) and brine solution (2 × 50.0 mL). The organic layers were dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. The crude product was purified by column chromatography using 10% ethyl acetate in hexane to obtain 1,2,3-tris({5-[(7-methyloctanoyl)oxy]pentyl})2-hydroxypropane-1,2,3-tricarboxylate[P-5] (4.5 g, 5.17 mmol) as a colorless, clear liquid. 1 H NMR(400MHz,CDCl3):δ 4.22(t,J=5.6Hz,2H),4.09(m,10H),2.89-2.77(dd,J=15.6Hz,4H),2.29(t,J=7.6Hz,6H), 1.64(m,16H),1.50(m,4H),1.49(m,6H),1.28(m,13H),1.15(m,6H),0.85(d,J=6.8Hz,18H).

[0735] TL1-14D-E5-7-6i (compound (36)) [ka] A stirred solution of 1,2,3-tris({5-[(7-methyloctanoyl)oxy]pentyl})2-hydroxypropane-1,2,3-tricarboxylate[P-5] (5.2 g, 5.97 mmol) in dichloromethane (61.2 mL, 956 mmol) was cooled to 0°C. 3-(dimethylamino)propanoic acid (2.8 g, 4 equivalents, 23.9 mmol) was added and the mixture was stirred for 10 minutes. {3-[cyano(ethyl)amino]propyl}dimethylazanium chloride (4.58 g, 4 equivalents, 23.9 mmol) was added at the same temperature, followed by the addition of N,N-dimethylpyridine-4-amine (729 mg, 5.97 mmol). The reaction mixture was stirred at room temperature for 16 hours. The progress of the reaction was monitored by ELSD / TLC. The reaction mixture was diluted with cold water (500.0 mL) and extracted with dichloromethane (2 × 150 mL). The combined organic layer was washed with fresh water (2 × 100 mL) and brine solution (2 × 100.0 mL). The organic layer was dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. The crude product was purified by column chromatography using 50% ethyl acetate in hexane to obtain [TL1-14D-E5-7-6i] (compound (36)) (1.85 g, 1.91 mmol) as a brown liquid mass. 1 H NMR(400MHz,CDCl3):δ 4.15(t,J=6.8Hz,2H),4.08(m,10H),3.31-3.27(d,J=15.6Hz,2H),3.23-3.19(d,J=15.6Hz,2H),2.58(t,J=7.2Hz,2H),2.48(t,J=7.2Hz, 2H),2.28(t,J=7.6Hz,6H),2.22(s,6H),1.69-1.5(m,18H),1.51(m,3H),1.42(m,6H),1.29(m,12H),1.14(m,6H),0.87(d,J=6.8Hz,18H).

[0736] LCMS analysis: purity 98.45%, calculated value C 53 H 95 NO 14 =969.68, measured value =970.68 (m / z, M+H + ).

[0737] Example 17. Synthesis protocol for M1266-480517 (TL1-12D-001) (Compound (33)) Synthesis scheme Q [ka] Intermediate [Q-3]: [ka] A stirred solution of 4-(tert-butoxy)-4-oxobutanoic acid [Q-1] (10 g, 0.0574 mol) and 2-(dimethylamino)ethane-1-ol [Q-2] (6.14 g, 0.0689 mol) in dichloromethane (100 mL) was cooled to 0°C. ({[3-(dimethylamino)propyl]imino}methylidene)(ethyl)amine hydrochloride (13.9 g, 0.0723 mol), followed by 4-(dimethylamino)pyridine-1-ium (1.41 g, 0.0115 mol), was added, and the reaction mixture was stirred at room temperature for 48 hours. The progress of the reaction was monitored by ELSD / TLC (SM was consumed). Water (50 mL) was added to the reaction mixture and extracted with DCM (3 × 50 mL). The resulting organic layer was dried over Na2SO4 and concentrated under reduced pressure. The crude product was purified by flash column chromatography (SiO2: 0-5% methanol in dichloromethane) to obtain the desired tert-butyl (2-(dimethylamino)ethyl) succinate [Q-3] (9.0 g, yield 63.91%) as a yellow liquid. 1 H-NMR(400MHz,CDCl3)-δ 4.16(t,J=6.0Hz,2H),2.59-2.41(m,6H),2.24(s,6H),1.41(s,9H)

[0738] Intermediate [Q-4]: [ka] To a stirred solution of tert-butyl(2-(dimethylamino)ethyl) succinate [Q-3] (9 g, 0.036 mol) in DCM (100 mL), trifluoroacetic acid (10 mL, 0.132 mol) was added at 0°C. The resulting reaction mixture was stirred at room temperature for 16 hours. The progress of the reaction mixture was monitored by ELSD / TLC (SM was consumed). The resulting reaction mixture was quenched with triethylamine to pH 7. The resulting organic layer was concentrated under reduced pressure, and the crude product was used directly in the next step.

[0739] ELSD analysis: Purity 99.15%, calculated C8H 15 NO4 = 189.10, Measured value = 189.80 (m / z, M+H) + ).

[0740] Intermediate [Q-7]: [ka] A stirred solution of octanoic acid [Q-6] (10 g, 69.3 mmol) and pentane-1,5-diol [Q-5] (6.5 g, 62.4 mmol) in dichloromethane (200 mL, 781 mmol) was cooled to 0°C. ({[3-(dimethylamino)propyl]imino}methylidene)(ethyl)amine hydrochloride (16.7 g, 87.4 mmol), followed by 4-(dimethylamino)pyridine-1-ium (1.71 g, 13.9 mmol), and the reaction mixture was stirred at room temperature for 48 hours. The progress of the reaction was monitored by ELSD / TLC (SM was consumed). Water (200 mL) was added to the reaction mixture and extracted with DCM (3 × 500 mL). The resulting organic layer was dried over Na2SO4, concentrated under reduced pressure, and the crude product was purified by flash column chromatography (SiO2: 0-10% ethyl acetate in hexane) to obtain the desired 5-hydroxypentyl octanoate [Q-7] (6.0 g, 26.0 mmol, yield 37.56%) as a yellow liquid. 1H-NMR(400MHz,CDCl3)-δ 4.10-4.07(m,2H),3.67(bs,2H),2.32-2.28(m,2H),1.71-1.58(m,4H),1.48-1.42(m,2H),1.36-1.29(m,9H),0.90-0.87(m,3H).

[0741] Intermediate [Q-9]: [ka] A stirred solution of 2-hydroxypropane-1,2,3-tricarboxylic acid [Q-8] (1.25 g, 6.51 mmol) and 5-hydroxypentyl octanoate [Q-7] (6 g, 26 mmol) in dichloromethane (50 mL, 781 mmol) was cooled to 0°C. ({[3-(dimethylamino)propyl]imino}methylidene)(ethyl)amine hydrochloride (4.99 g, 26 mmol), followed by 4-(dimethylamino)pyridine-1-ium (802 mg, 6.51 mmol), and the reaction mixture was stirred at room temperature for 48 hours. The progress of the reaction was monitored by ELSD / TLC (SM was consumed). Water (50 mL) was added to the reaction mixture and extracted with DCM (3 × 100 mL). The resulting organic layer was dried over Na2SO4, concentrated under reduced pressure, and the crude product was purified by flash column chromatography (SiO2: 0-30% ethyl acetate in hexane) to obtain the desired tris(5-(octanoyloxy)pentyl)2-hydroxypropane-1,2,3-tricarboxylate[Q-9] (2.1 g, 2.53 mmol, yield 38.9%) as a yellow liquid. 1 H-NMR(400MHz,CDCl3)-δ 4.23-4.20(m,2H),4.10-4.04(m,10H),2.89-2.77(dd,J=15.6,19.2Hz,4H),2.30-2 .26(m,6H),1.75-1.56(m,18H),1.45-1.36(m,6H),1.29-1.28(m,24H),0.88(m,9H).

[0742] ELSD analysis: Purity 97.90%, calculated value C 45 H 80O 13 =828.56, measured value =829.35 (m / z, M+H + ).

[0743] TL1-12D-001 (compound (33)) [ka] A stirred solution of 4-[2-(dimethylamino)ethoxy]-4-oxobutanoic acid [Q-4] (1.92 g, 10.1 mmol) and 1,2,3-tris[5-(octanoyloxy)pentyl]2-hydroxypropane-1,2,3-tricarboxylate [Q-9] (2.1 g, 2.53 mmol) in dichloromethane (15 mL, 234 mmol) was cooled to 0°C. ({[3-(dimethylamino)propyl]imino}methylidene)(ethyl)amine hydrochloride (1.94 g, 10.1 mmol), followed by 4-(dimethylamino)pyridine-1-ium (312 mg, 2.53 mmol), and the reaction mixture was stirred at room temperature for 48 hours. The progress of the reaction was monitored by ELSD / TLC (SM was consumed). Water (50 mL) was added to the reaction mixture and extracted with DCM (3 × 100 mL). The resulting organic layer was dried over Na₂SO₄ and concentrated under reduced pressure. The crude product was purified by flash column chromatography (SiO₂: 0-100% ethyl acetate in hexane) to obtain the desired product [TL1-12D-001] (compound (33)) (115 mg, yield 4.54%) as a clear liquid. 1 H-NMR(400MHz,DMSO-d6)-δ 4.08-3.97(m,14H),3.17-3.13(d,J=15.6Hz,2H),3.10-3.06(d,J=15.2Hz,2H),2.46-2.43(m,6 H),2.27-2.24(m,6H),2.14(s,6H),1.58-1.48(m,18H),1.35-1.23(m,30H),0.86-0.83(m,9H).

[0744] ELSD analysis: Purity 99.90%, calculated value C 53 H 93 NO 16=999.65, measured value =1000.50 (m / z, M+H + ).

[0745] Example 18. Synthesis protocol for M1266-497687 (TL1-10D-006) (Compound (27)) Synthesis scheme R [ka] Intermediate [R-3]: [ka] A stirred solution of 4-(tert-butoxy)-4-oxobutanoic acid [R-1] (1.5 g, 0.0086 mol) and 2-(dimethylamino)ethane-1-ol [R-2] (0.92 g, 0.0103 mol) in dichloromethane (20 mL) was cooled to 0°C. ({[3-(dimethylamino)propyl]imino}methylidene)(ethyl)amine hydrochloride (3.30 g, 0.0172 mol), followed by 4-(dimethylamino)pyridine-1-ium (1.0 g, 0.0086 mol), was added, and the reaction mixture was stirred at room temperature for 16 hours. The progress of the reaction was monitored by ELSD / TLC (SM was consumed). Water (20 mL) was added to the reaction mixture and extracted with DCM (3 × 50 mL). The resulting organic layer was dried over Na2SO4 and concentrated under reduced pressure. The crude product was purified by flash column chromatography (SiO2: 0-5% methanol in dichloromethane) to obtain the desired tert-butyl(2-(dimethylamino)ethyl) succinate [R-3] (1.5 g, yield 71%) as a pale yellow liquid. 1 H-NMR(400MHz,CDCl3)-δ 4.16(t,J=6.0Hz,2H),2.59-2.48(m,6H),2.24(s,6H),1.41(s,9H)

[0746] Intermediate [R-4]: [ka] Trifluoroacetic acid (1.4 g, 0.012 mol) was added at 0°C to a stirred solution of tert-butyl(2-(dimethylamino)ethyl) succinate [R-3] (1.5 g, 0.006 mol) in DCM (15 mL). The resulting reaction mixture was stirred at room temperature for 16 hours. The progress of the reaction mixture was monitored by ELSD / TLC (SM was consumed). The resulting reaction mixture was quenched with triethylamine to pH 7. The resulting organic layer was concentrated under reduced pressure, and the crude product was used directly in the next step.

[0747] ELSD analysis: Purity 99.15%, calculated C8H 15 NO4 = 189.10, Measured value = 189.80 (m / z, M+H) + ).

[0748] Intermediate [R-7]: [ka] A stirred solution of 5-methylhexanoic acid [R-6] (10 g, 76.8 mmol) and pentane-1,5-diol [R-5] (8 g, 76.8 mmol) in dichloromethane (150 mL) was cooled to 0°C, and EDC.HCl (18.6 g, 96.8 mmol), followed by 4-(dimethylamino)pyridine-1-ium (1.89 g, 15.4 mmol), was added and the mixture was stirred at room temperature for 16 hours. The progress of the reaction was monitored by TLC (SM was consumed). Water (100 mL) was added to the reaction mixture and extracted with DCM (3 × 100 mL). The resulting organic layer was dried over Na2SO4 and concentrated under reduced pressure to obtain the crude product, which was purified by flash column chromatography using 0-20% ethyl acetate in n-hexane to obtain the pure product 5-hydroxypentyl 5-methylhexanoate [R-7] (6.1 g, 37%) as a pale yellow liquid. 1H-NMR(400MHz,CDCl3)-δ 4.07(t,J=6.4Hz,2H),3.65(t,J=6.4Hz,2H),2.27(t,J=7.2Hz,2H),1.69- 1.53(m,8H),1.51-1.39(m,2H),1.24-1.16(m,2H),0.87(d,J=6.8Hz,6H).

[0749] Intermediate [R-9]: [ka] A stirred solution of 5-hydroxypentyl 5-methylhexanoate [R-7] (6 g, 27.7 mmol) and 2-hydroxypropane-1,2,3-tricarboxylic acid [R-8] (1.33 g, 6.93 mmol) in dichloromethane (150 mL) was cooled to 0°C, and EDC.HCl (5.32 g, 27.7 mmol), followed by 4-(dimethylamino)pyridine-1-ium (854 mg, 6.93 mmol), was added and the mixture was stirred at room temperature for 48 hours. The progress of the reaction was monitored by TLC (SM was consumed). Water (100 mL) was added to the reaction mixture and extracted with DCM (3 × 200 mL). The resulting organic layer was dried over Na2SO4 and concentrated under reduced pressure to obtain the crude product, which was purified by flash column chromatography (0-30% ethyl acetate in n-hexane) to obtain the pure product 1,2,3-tris({5-[(5-methylhexanoyl)oxy]pentyl})2-hydroxypropane-1,2,3-tricarboxylate[R-9] (1.9 g, 35%) as a pale yellow liquid. 1 H-NMR(400MHz,CDCl3)-δ 4.21(t,J=6.4Hz,2H),4.10-4.04(m,10H),2.89-2.77(dd,J=15.6,19.2Hz,4H),2.27(t,J=7 .6Hz,6H),1.68-1.51(m,22H),1.41-1.38(m,6H),1.21-1.16(m,6H),0.88(d,J=6.4Hz,18H).

[0750] JC-TL1-10D-006 (Compound (27)) [ka] A stirred solution of 1,2,3-tris({5-[(5-methylhexanoyl)oxy]pentyl})2-hydroxypropane-1,2,3-tricarboxylate[R-9] (1.9 g, 2.41 mmol) and 4-[2-(dimethylamino)ethoxy]-4-oxobutanoic acid[R-4] (1.83 g, 9.66 mmol) in dichloromethane (20 mL) was cooled to 0°C, and EDC.HCl (1.85 g, 9.66 mmol), followed by 4-(dimethylamino)pyridine-1-ium (0.247 g, 2.41 mol), was added and the mixture was stirred at room temperature for 48 hours. The progress of the reaction was monitored by TLC (SM was consumed). Water (20 mL) was added to the reaction mixture and extracted with DCM (3 × 50 mL). The obtained organic layer was dried over Na2SO4 and concentrated under reduced pressure to obtain the crude product, which was purified by flash column chromatography (0-100% ethyl acetate in n-hexane) to obtain the pure product [JC-TL1-10D-006] (compound (27)) (0.5 g, 22%) as a brown liquid. 1 H-NMR(400MHz,CDCl3)-δ 4.19(t,J=4.8Hz,2H),4.14(t,J=6.8Hz,2H),4.09-4.04(m,10H),3.30-3.19(dd,J=15.6,12.8Hz,4H),2.63-2.62(m,4H),2.60-2.5 9(br,2H),2.30-2.29(m,4H),2.27-2.25(m,7H),1.71-1.51(m,21H),1.44-1.36(m,6H),1.21-1.12(m,6H),0.88(d,J=6.4Hz,18H).

[0751] ELSD analysis: Purity 99.89%, calculated value C 50 H 87 NO 16 =957.60, measured value =958.45 (m / z, M+H + ).

[0752] Example 19. Synthesis protocol for M1266-499030 (TL1-14D-003) (Compound (24)) Synthesis scheme S [ka] Intermediate [S-3]: [ka] A stirred solution of 4-(tert-butoxy)-4-oxobutanoic acid [S-1] (10 g, 0.0574 mol) and 2-(dimethylamino)ethane-1-ol [S-2] (6.14 g, 0.0689 mmol) in dichloromethane (100 mL) was cooled to 0°C. ({[3-(dimethylamino)propyl]imino}methylidene)(ethyl)amine hydrochloride (13.9 g, 0.0723 mol), followed by 4-(dimethylamino)pyridine-1-ium (1.41 g, 0.0115 mol), was added, and the reaction mixture was stirred at room temperature for 48 hours. The progress of the reaction was monitored by ELSD / TLC (SM was consumed). Water (50 mL) was added to the reaction mixture and extracted with DCM (3 × 100 mL). The resulting organic layer was dried over Na2SO4, concentrated under reduced pressure, and the crude product was purified by flash column chromatography (SiO2: 0-5% methanol in dichloromethane) to obtain the desired tert-butyl(2-(dimethylamino)ethyl) succinate [S-3] (9.0 g, yield 63.91%) as a pale yellow liquid.

[0753] result: 1 H-NMR(400MHz,CDCl3)-δ 4.16(t,J=6.0Hz,2H),2.59-2.41(m,6H),2.24(s,6H),1.41(s,9H)

[0754] Intermediate [S-4]: [ka] To a stirred solution of tert-butyl(2-(dimethylamino)ethyl) succinate [S-3] (9 g, 0.036 mol) in DCM (100 mL), trifluoroacetic acid (10 mL, 0.132 mol) was added at 0°C. The resulting reaction mixture was stirred at room temperature for 16 hours. The progress of the reaction mixture was monitored by ELSD / TLC (SM was consumed). The resulting reaction mixture was quenched with triethylamine to pH 7. The resulting organic layer was concentrated under reduced pressure, and the crude product was used directly in the next step.

[0755] result: ELSD analysis: Purity 99.15%, calculated C8H 15 NO4 = 189.10, Measured value = 189.80 (m / z, M+H) + ).

[0756] Intermediate [S-7]: [ka] A stirred solution of decanoic acid [S-6] (10 g, 58 mmol) and pentane-1,5-diol [5] (5.44 g, 52.2 mmol) in dichloromethane (200 mL) was cooled to 0°C. ({[3-(dimethylamino)propyl]imino}methylidene)(ethyl)amine hydrochloride (14 g, 73.1 mmol), followed by 4-(dimethylamino)pyridine-1-ium (1.43 g, 11.6 mmol), and the reaction mixture was stirred at room temperature for 16 hours. The progress of the reaction was monitored by TLC (SM was consumed). Water (100 mL) was added to the reaction mixture and extracted with DCM (3 × 300 mL). The resulting organic layer was dried over Na2SO4, concentrated under reduced pressure, and the crude product was purified by flash column chromatography (SiO2: 0-10% ethyl acetate in hexane) to obtain the desired 5-hydroxypentyldecanoate [S-7] (3.6 g, yield 24%) as a yellow liquid.

[0757] result: 1H-NMR(400MHz,CDCl3)-δ 4.09(t,J=6.4Hz,2H),3.67(q,J=6.0Hz,2H),2.30(t,J=7.6Hz,2H),1.70-1.56(m,6H),1.49-1.43(m,2H),1.31-1.23(m,12H),0.91(m,3H).

[0758] ELSD analysis: Purity 99.43%, calculated value C 15 H 30 O3 = 258.22, measured value = 259.15 (m / z, M+H + ).

[0759] Intermediate [S-9]: [ka] A stirred solution of 2-hydroxypropane-1,2,3-tricarboxylic acid [S-8] (651 mg, 3.39 mmol) and 5-hydroxypentyldecanoate [S-7] (3.5 g, 13.5 mmol) in dichloromethane (50 mL) was cooled to 0°C. ({[3-(dimethylamino)propyl]imino}methylidene)(ethyl)amine hydrochloride (2.6 g, 13.5 mmol), followed by 4-(dimethylamino)pyridine-1-ium (417 mg, 3.39 mmol), and the reaction mixture was stirred at room temperature for 48 hours. The progress of the reaction was monitored by TLC (SM was consumed). Water (20 mL) was added to the reaction mixture and extracted with DCM (3 × 100 mL). The resulting organic layer was dried over Na2SO4 and concentrated under reduced pressure. The crude product was purified by flash column chromatography (SiO2: 0-10% ethyl acetate in hexane) to obtain the desired 1,2,3-tris[5-(decanoyloxy)pentyl]2-hydroxypropane-1,2,3-tricarboxylate[S-9] (1.97 g, 2.16 mmol) as a pale yellow liquid. 1H-NMR(400MHz,CDCl3)-δ 4.22(t,J=6.4Hz,2H),4.11-4.04(m,10H),2.88(dd,J=15.6,19.2Hz,4H),2.28(t,J=7.6H z,6H),1.73-1.59(m,18H),1.43-1.36(m,6H),1.29-1.26(m,36H),0.88(d,J=6.8Hz,9H).

[0760] ELSD analysis: Purity 98.31%, calculated value C 51 H 92 O 13 =912.65, measured value =935.90 (m / z, M+Na + ).

[0761] JC-TL1-14D-003 (compound (24)) [ka] A stirred solution of 4-[2-(dimethylamino)ethoxy]-4-oxobutanoic acid [S-4] (1.51 g, 8.32 mmol) and 1,2,3-tris[5-(decanoyloxy)pentyl]2-hydroxypropane-1,2,3-tricarboxylate [S-9] (1.9 g, 2.41 mmol) in dichloromethane (50 mL) was cooled to 0°C. ({[3-(dimethylamino)propyl]imino}methylidene)(ethyl)amine hydrochloride (1.6 g, 8.32 mmol), followed by 4-(dimethylamino)pyridine-1-ium (254 mg, 2.08 mmol), was added, and the reaction mixture was stirred at room temperature for 48 hours. The progress of the reaction was monitored by ELSD / TLC (SM was consumed). Water (50 mL) was added to the reaction mixture and extracted with DCM (3 × 100 mL). The resulting organic layer was dried over Na2SO4, concentrated under reduced pressure, and the crude product was purified by flash column chromatography (SiO2: 0-100% ethyl acetate in hexane) to obtain the desired product [JC-TL1-14D-003] (compound (24)) (0.6 g, 26%) as a yellow oil. 1H-NMR(400MHz,CDCl3)-δ 4.20(t,J=5.6Hz,2H),4.14(t,J=6.8Hz,2H),4.09-4.03(m,10H),3.28(d,J=15.6Hz,2H),3.21(d,J=15.6Hz,2H),2.6 5-2.60(m,6H),2.30-2.26(m,12H),1.71-1.56(m,18H),1.44-1.36(m,6H),1.29-1.25(m,36H),0.87(d,J=6.8Hz,9H).

[0762] ELSD analysis: Purity 99.72%, calculated value C 59 H 105 NO 16 =1083.74, measured value =1084.50 (m / z, M+H + ).

[0763] Example 20. Synthesis protocol for M1266-507720 (TL1-12D-002) (Compound (35)) Synthesis scheme T [ka] Intermediate [T-3]: [ka] A stirred solution of 4-(tert-butoxy)-4-oxobutanoic acid [T-1] (10 g, 0.0574 mol) and 2-(dimethylamino)ethane-1-ol [T-2] (6.14 g, 0.0689 mmol) in dichloromethane (100 mL) was cooled to 0°C. ({[3-(dimethylamino)propyl]imino}methylidene)(ethyl)amine hydrochl...

Claims

1. Compounds having the structure shown in the following formula: Formula (A2): 【Chemistry 1】 or a pharmaceutically acceptable salt thereof (in the formula, X is independently either O or NH; R １ 、R ２ 、and R ３ each is independently C ４ -C ３０ alkyl, C ４ -C ３０ alkenyl, C ４ -C ３０ alkynyl, or C ４ -C ３０ heteroalkyl; A is, 【Chemistry 2】 It is a substructure selected from, B is independently an ionizable nitrogen-containing group or a permanently charged nitrogen group; m is an integer between 2 and 10; n is an integer between 2 and 10; L １ is a carbonyl, ester, or amide; L ２ C ２ ~C １０ Alkylene or C ２ ~C １０ It is alkenylene; Ar is a halogen, OCH ３ , and CH ３ A phenylene comprising any 1 to 4 substituents independently selected from; R ４ C １ ~C １０ It is alkylene; Here, A is R １ , R ２ , and R ３ Each of them independently contains a disulfide bond, or 【Transformation 3】 C has the structure ４ ~C ３０ In the case of a heteroalkyl group, it is the only substructure (a4), L ３ is OC(O), CO ２ , or (O)CO; o is an integer between 2 and 5; R ５ C ４ ~C ２４ Alkyl; or L ３ is OC(O), CO ２ , or (O)CO; o is an integer from 6 to 12; R ５ C １ ~C ６ (It is alkyl); or Formula (A1): 【Chemistry 4】 or a pharmaceutically acceptable salt thereof (in the formula, X is independently either O or NH; R １ , R ２ , and R ３ Each of them independently, C ４ ~C ３０ Alkyl, C ４ ~C ３０ Alkenil, C ４ ~C ３０ Alkinyl, or C ４ ~C ３０ It is heteroalkyl; A is, 【Transformation 5】 It is a substructure selected from, B is independently an ionizable nitrogen-containing group or a permanently charged nitrogen group; m is an integer between 2 and 10; n is an integer between 2 and 10; L １ is a carbonyl, ester, or amide; L ２ C ２ ~C １０ Alkylene or C ２ ~C １０ It is alkenylene; Ar is a halogen, OCH ３ , and CH ３ A phenylene comprising any 1 to 4 substituents independently selected from; R ４ C １ ~C １０ It is alkylene; Here, A is R １ , R ２ , and R ３ Each of them independently contains a disulfide bond, or 【Transformation 6】 C has the structure ４ ~C ３０ In the case of a heteroalkyl group, it is the only substructure (a4), L ３ is OC(O), CO ２ , or (O)CO; o is an integer between 2 and 5; R ５ C ４ ~C ２４ Alkyl; or L ３ is OC(O), CO ２ , or (O)CO; o is an integer from 6 to 12; R ５ C １ ~C ５ (It is alkyl); or Formula (A): 【Transformation 7】 or a pharmaceutically acceptable salt thereof (in the formula, X is independently either O or NH; R １ , R ２ , and R ３ Each of them independently, C ４ ~C ３０ Alkyl, C ４ ~C ３０ Alkenil, C ４ ~C ３０ Alkinyl, or C ４ ~C ３０ It is heteroalkyl; A is, 【Transformation 8】 It is a substructure selected from, B is independently an ionizable nitrogen-containing group or a permanently charged nitrogen group; m is an integer between 2 and 10; n is an integer between 2 and 10; L １ is a carbonyl, ester, or amide; L ２ is C ２ to C １０ alkylene or C ２ to C １０ alkenylene; Ar is a halogen, OCH ３ , and CH ３ A phenylene comprising any 1 to 4 substituents independently selected from; R ４ C ２ ~C １０ It is alkylene; Here, A is R １ , R ２ , and R ３ each independently contains a disulfide bond or 【Chemistry 9】 C has the structure ４ ~C ３０ In the case of a heteroalkyl group, it is the only substructure (a4), L ３ is OC(O), CO ２ , or (O)CO; o is an integer between 2 and 5; R ５ C ４ ~C ２４ Alkyl; or L ３ is OC(O), CO ２ , or (O)CO; o is an integer from 6 to 12; R ５ C １ ~C ５ (It is alkyl.)

2. A compound according to claim 1 having the structure of formula (I): 【Chemistry 10】 or a pharmaceutically acceptable salt thereof (in the formula, R １ , R ２ , and R ３ Each of them independently, C ６ ~C ３０ Alkyl, C ６ ~C ３０ Alkenil, C ６ ~C ３０ Alkinyl, or C ４ ~C ３０ It is heteroalkyl; m is an integer between 2 and 10; n is an integer between 2 and 10; B is independently an ionizable nitrogen-containing group or a permanently charged nitrogen group.

3. The compound according to claim 1, having the structure of formula (III): 【Chemistry 11】 or a pharmaceutically acceptable salt thereof (in the formula, R １ , R ２ , and R ３ Each of them independently, C ６ ~C ３０ Alkyl, C ６ ~C ３０ Alkenil, C ６ ~C ３０ Alkinyl, or C ４ ~C ３０ It is heteroalkyl; R ４ C ２ ~C １０ It is alkylene; B is an independently ionizable nitrogen-containing group.

4. The compound according to claim 1, having the structure of formula (IV): 【Chemistry 12】 or a pharmaceutically acceptable salt thereof (in the formula, R １ , R ２ , and R ３ Each of them independently, C ６ ~C ３０ Alkyl, C ６ ~C ３０ Alkenil, C ６ ~C ３０ Alkinyl, or C ４ ~C ３０ It is heteroalkyl; m is an integer between 2 and 10; n is an integer between 2 and 10; B is an independently ionizable nitrogen-containing group.

5. The compound according to claim 1, having the structure shown in the following formula: (a) Formula (V1): 【Chemistry 13】 or a pharmaceutically acceptable salt thereof (in the formula, n is an integer of 2, 3, 4, 5, 6, or 7; B is an independently ionizable nitrogen-containing group); or (b) Formula (V): 【Chemistry 14】 or a pharmaceutically acceptable salt thereof (in the formula, n is an integer of 2, 3, or 4; B is an independently ionizable nitrogen-containing group.

6. The compound according to claim 1, having the structure of formula (VI): 【Chemistry 15】 or a pharmaceutically acceptable salt thereof (in the formula, R １ , R ２ , and R ３ Each of them independently contains a disulfide group C ６ ~C ３０ It is heteroalkyl; n is an integer of 2, 3, or 4; B is an independently ionizable nitrogen-containing group.

7. Formula related to formula (VII): 【Chemistry 16】 A compound having, or a pharmaceutically acceptable salt thereof (wherein, X is independently either O or NH; R １ , R ２ , and R ３ Each of them independently, C ４ ~C ３０ Alkyl, C ４ ~C ３０ Alkenil, C ４ ~C ３０ Alkinyl, or C ４ ~C ３０ It is heteroalkyl; Each m is an integer between 2 and 10; Each L ４ is a carbonyl, ester, or amide; Z is -(CH ２ ) ｑ１ -N-(CH ２ ) ｑ２ - and q1 and q2 are independently integers between 2 and 10; or Z is C ６ H ３ -Z １ And here, Z １ is, -CH ２ (CH ２ ) ｑ The B portion is covalently bonded to a carbonyl, ester, or amide; q is an integer between 1 and 9; B is an independently ionizable nitrogen-containing group.

8. (a) Compounds 41-64 and 90, or pharmaceutically acceptable salts thereof: Table 1 Table 2 Table 3 Table 4 Table 5 Table 6 Table 7 (b) Compounds 91-111, or pharmaceutically acceptable salts thereof: Table 8 Table 9 Table 10 Table 11 Table 12 Table 13 (c) Compounds 112-117 or 126-129, or pharmaceutically acceptable salts thereof: Table 14 Table 15 Table 16 (d) Compounds 118-125 or pharmaceutically acceptable salts thereof: Table 17 Table 18 Table 19 A compound selected from the group.

9. A composition comprising nucleic acids encapsulated within liposomes, wherein the liposomes comprise a cationic lipid which is a compound according to any one of claims 1 to 8.

10. A method for delivering a composition containing mRNA encoding a protein or polypeptide, encapsulated within a liposome, to the lung, intranasal cavity, or intramuscular muscle, wherein the liposome comprises one or more cationic lipids, optionally one or more non-cationic lipids, optionally one or more cholesterol-based lipids, and optionally one or more PEG-modified lipids, and at least one cationic lipid having a structure relating to formula (B): 【Chemistry 17】 A compound having, or a pharmaceutically acceptable salt thereof, wherein, Each n is independently either 0 or 1; X １Ａ Independently, O or NR １Ａ And; R １Ａ is H or C １ ~C ６ It is alkyl, X １Ｂ is a covalent bond, C(O), CH ２ CO ２ , or CH ２ It is C(O); X ２Ａ and X ２Ｂ One of them is O, and the other is a covalent bond; X ３Ａ and X ３Ｂ One of them is O, and the other is a covalent bond; X ４Ａ and X ４Ｂ One of them is O, and the other is a covalent bond; R １ Independently, L １ -B １ , C ６ ~C ３０ Alkyl, C ６ ~C ３０ Alkenyl, or C ６ ~C ３０ It is alkinyl; R ２ Independently, L ２ -B ２ , C ６ ~C ３０ Alkyl, C ６ ~C ３０ Alkenyl, or C ６ ~C ３０ It is alkinyl; R ３ Independently, L ３ -B ３ , C ６ ~C ３０ Alkyl, C ６ ~C ３０ Alkenyl, or C ６ ~C ３０ It is alkinyl; R ４ Independently, L ４ -B ４ , C ６ ~C ３０ Alkyl, C ６ ~C ３０ Alkenyl, or C ６ ~C ３０ It is alkinyl; L １ , L ２ , L ３ , and L ４ Each of them is independent, C １ ~C ３０ Alkylene; C ２ ~C ３０ Alkenylene; or C ２ ~C ３０ It is alkynylene; B １ , B ２ , B ３ , and B ４ Each of them is independently an ionizable nitrogen-containing group, The method wherein the cationic lipid comprises at least one ionizable nitrogen-containing group.

11. A method for delivering a composition containing nucleic acid encapsulated in liposomes to the lungs, nasal cavity, or muscle, wherein the liposomes have a structure relating to formula (B): [Chemistry 18] A compound having, or a cationic lipid which is a pharmaceutically acceptable salt thereof, in the formula, Each n is independently either 0 or 1; X １Ａ Independently, O or NR １Ａ And; R １Ａ is H or C １ ~C ６ It is alkyl, X １Ｂ is a covalent bond, C(O), CH ２ CO ２ , or CH ２ It is C(O); X ２Ａ and X ２Ｂ One of them is O, and the other is a covalent bond; X ３Ａ and X ３Ｂ One of them is O, and the other is a covalent bond; X ４Ａ and X ４Ｂ One of them is O, and the other is a covalent bond; R １ Independently, L １ -B １ , C ６ ~C ３０ Alkyl, C ６ ~C ３０ Alkenyl, or C ６ ~C ３０ It is alkinyl; R ２ Independently, L ２ -B ２ , C ６ ~C ３０ Alkyl, C ６ ~C ３０ Alkenyl, or C ６ ~C ３０ It is alkinyl; R ３ Independently, L ３ -B ３ , C ６ ~C ３０ Alkyl, C ６ ~C ３０ Alkenyl, or C ６ ~C ３０ It is alkinyl; R ４ Independently, L ４ -B ４ , C ６ ~C ３０ Alkyl, C ６ ~C ３０ Alkenyl, or C ６ ~C ３０ It is alkinyl; L １ , L ２ , L ３ , and L ４ Each of them is independent, C １ ~C ３０ Alkylene; C ２ ~C ３０ Alkenylene; or C ２ ~C ３０ It is alkynylene; B １ , B ２ , B ３ , and B ４ Each of them is independently an ionizable nitrogen-containing group, The method wherein the cationic lipid comprises at least one ionizable nitrogen-containing group.

12. The aforementioned compounds are (a) compounds (65) to (89), or pharmaceutically acceptable salts thereof: Table 20 Table 21 Table 22 Table 23 Table 24 Table 25 Table 26 (b) Compounds 118-125, or pharmaceutically acceptable salts thereof: Table 27 Table 28 Table 29 The method according to claim 10 or 11, selected from the group consisting of the following.

13. The method according to any one of claims 10 to 12, wherein the delivery is intranasal.

14. The method according to any one of claims 10 to 12, wherein the delivery is intramuscular.

15. The method according to any one of claims 10 to 12, wherein the delivery is intrapulmonary.

Images