Nanomaterial

Abstract

Lipid nanoparticle compositions for the delivery of nucleic acids are provided.SOLUTION: In various embodiments, the lipid nanoparticle comprises an ionizable lipid of Formula (I). Also provided are methods of using such lipid nanoparticle compositions to achieve targeted delivery of therapeutic cargo without the need for a targeting ligand.SELECTED DRAWING: None

Description

[Technical Field]

[0001] Information about related applications This application was filed on January 9, 2020, the entire contents of which are incorporated herein by reference. This application claims the benefit of U.S. Provisional Patent Application No. 62 / 958,876. [Background technology]

[0002] Field This application relates to the fields of chemistry, biology, and medicine. Disclosed herein are drug delivery systems and methods of their use. Disclosed are nanoparticle compositions for the delivery of nucleic acids to cells.

[0003] explanation Hepatocytes help maintain homeostasis and produce many secretory proteins, thus It has been suggested that nanoparticles carrying RNA are targeted to liver cells. However, systemic delivery to hepatocytes without a targeting ligand remains challenging. This is a difficult challenge. Summary of the Invention

[0004] Some embodiments described herein include compounds of formula (I):

[0005] [ka] Regarding the compound (In the formula, R 1 is C9~C 20 Alkyl or C9-C with 1-3 units of unsaturation 20 Arke Nil; X 1 and X 2 are each independently absent, -O-, or -NR2 - and

[0006] [ka] where each R 2 are independently hydrogen or C1-C6 alkyl; each a is independently an integer between 1 and 6; X 3 and X 4 are each independently absent or contain one or two C1~ 4-8 membered heterocyclyl optionally substituted with C6 alkyl group, 1 or 2 5-6 membered heteroaryl optionally substituted with C1-C6 alkyl group, 1 or 5-6 membered aryl optionally substituted with two C1-C6 alkyl groups, 1 or 4- to 7-membered cycloalkyl optionally substituted with two C1-C6 alkyl groups, -O -, and -NR 3 -, wherein each R 3 is a hydrogen atom or C1 ∼C6 alkyl, where X 1 -X 2 -X 3 -X 4 is an oxygen-oxygen bond, oxygen-nitrogen bond Contains no elementary or nitrogen-nitrogen bonds; X 5 Ha-(CH2) b where b is an integer between 0 and 6; X 6 is hydrogen, C1-C6 alkyl, one or two C1-C6 alkyl groups, if necessary optionally substituted 5-6 membered heteroaryl, or -NR 4 R 5 where R 4 and R 5 are each independently hydrogen or C1-C6 alkyl; or alternatively In fact, R4 and R 5 together with the nitrogen to which they are attached, form one or two C Form a 4-7 membered heterocyclyl optionally substituted with a 1-C6 alkyl group, and heterocyclyl requires an additional heteroatom selected from oxygen, sulfur, and nitrogen. Include as needed; each X 7 are independently hydrogen, hydroxyl, or -NR 6 R 7 where R 6 oh Yobi R 7 are each independently hydrogen or C1-C6 alkyl; or alternatively , R 6 and R 7 These, together with the nitrogen to which they are attached, form one or two C1~ Form a 4- to 7-membered heterocyclyl optionally substituted with a C6 alkyl group, wherein Heterocyclyl requires an additional heteroatom selected from oxygen, sulfur, and nitrogen. Includes accordingly; X 1 , X 2 , X 3 , X 4 , and X 5 At least one of the following is present; A 1 and A 2 are independently C5 to C 12 Haloalkyl, C5-C 12 Al Kenyl, C5-C 12 Alkynyl, (C5-C 12 Alkoxy)-(CH2) n2 -, 1 or two halo groups, C1-C6 alkyl groups, C1-C6 haloalkyl groups, or C1 Optionally ring-substituted with C-C alkoxy groups (C-C 10 Aryl)-(CH2) n3- and optionally ring-substituted with one or two C1-C6 alkyl groups (C 3~C8 cycloalkyl)-(CH2) n4 - selected from the group consisting of; Alternatively, A 1 and A 2 together with the atoms to which they are attached form one or two C4~C 10 Forms 5-6 membered cyclic acetals substituted with alkyl groups; n1, n2, and n3 are each independently an integer between 1 and 4; n4 is an integer between 0 and 4).

[0007] Some embodiments include lipids comprising an ionizable lipid; a phospholipid; a polyethylene glycol; Lipid nanoparticle compositions containing ethylene glycol-lipid; cholesterol; and optionally nucleic acid. Further embodiments feature compositions comprising: a method for delivering a nucleic acid to a subject in need thereof; The present invention is directed to a method comprising administering a lipid nanoparticle composition to a subject in need thereof. do.

[0008] These and other embodiments are described in more detail below. [Brief explanation of the drawings]

[0009] [Figure 1] FIG. 1 illustrates a reaction scheme for preparing compounds of formula (I). [Figure 2-1] 1 is a table summarizing the structures of the compounds of Examples 1 to 148. [Figure 2-2] 1 is a table summarizing the structures of the compounds of Examples 1 to 148. [Figure 2-3] 1 is a table summarizing the structures of the compounds of Examples 1 to 148. [Figure 2-4] 1 is a table summarizing the structures of the compounds of Examples 1 to 148. [Figure 2-5]1 is a table summarizing the structures of the compounds of Examples 1 to 148. [Figure 2-6] 1 is a table summarizing the structures of the compounds of Examples 1 to 148. [Figure 2-7] 1 is a table summarizing the structures of the compounds of Examples 1 to 148. [Figure 2-8] 1 is a table summarizing the structures of the compounds of Examples 1 to 148. [Figure 2-9] 1 is a table summarizing the structures of the compounds of Examples 1 to 148. [Figure 2-10] 1 is a table summarizing the structures of the compounds of Examples 1 to 148. [Figure 2-11] 1 is a table summarizing the structures of the compounds of Examples 1 to 148. [Figure 2-12] 1 is a table summarizing the structures of the compounds of Examples 1 to 148. [Figure 2-13] 1 is a table summarizing the structures of the compounds of Examples 1 to 148. [Figure 2-14] 1 is a table summarizing the structures of the compounds of Examples 1 to 148. [Figure 2-15] 1 is a table summarizing the structures of the compounds of Examples 1 to 148. [Figure 2-16] 1 is a table summarizing the structures of the compounds of Examples 1 to 148. [Figure 2-17] 1 is a table summarizing the structures of the compounds of Examples 1 to 148. [Figure 2-18] 1 is a table summarizing the structures of the compounds of Examples 1 to 148. [Figure 2-19] 1 is a table summarizing the structures of the compounds of Examples 1 to 148. [Figure 2-20] 1 is a table summarizing the structures of the compounds of Examples 1 to 148. [Figure 2-21] 1 is a table summarizing the structures of the compounds of Examples 1 to 148. [Figure 2-22] 1 is a table summarizing the structures of the compounds of Examples 1 to 148. [Figure 2-23] 1 is a table summarizing the structures of the compounds of Examples 1 to 148. [Figure 2-24] 1 is a table summarizing the structures of the compounds of Examples 1 to 148. [Figure 2-25] 1 is a table summarizing the structures of the compounds of Examples 1 to 148. [Figure 2-26] 1 is a table summarizing the structures of the compounds of Examples 1 to 148. [Figure 2-27] 1 is a table summarizing the structures of the compounds of Examples 1 to 148. [Figure 2-28] 1 is a table summarizing the structures of the compounds of Examples 1 to 148. [Figure 2-29] 1 is a table summarizing the structures of the compounds of Examples 1 to 148. [Figure 2-30] 1 is a table summarizing the structures of the compounds of Examples 1 to 148. [Figure 2-31] 1 is a table summarizing the structures of the compounds of Examples 1 to 148. [Figure 2-32] 1 is a table summarizing the structures of the compounds of Examples 1 to 148. [Figure 2-33] 1 is a table summarizing the structures of the compounds of Examples 1 to 148. DETAILED DESCRIPTION OF THE INVENTION

[0010] definition Unless otherwise defined, all technical and scientific terms used herein are understood to be of ordinary skill in the art. All patents, publications, and other materials referenced herein have the same meaning as commonly understood by those skilled in the art. Applications, published applications, and other publications are incorporated by reference in their entirety unless otherwise indicated. In the event that there is a plurality of definitions for terms herein, the definitions shall be expressly incorporated herein. Unless otherwise indicated, the definitions in this section take precedence.

[0011] As used herein, any "R" or "X" group, including but not limited to, , R 1 , R 2 , R 3 , R 4 , R 5 , R 6 , R 7 , X1 , X 2 , X 3 , X 4 , X 5 , X 6 , X 7 , A 1 , and A 2 represents a substituent that can be attached to the indicated atom. The R, X, and A groups may be collectively referred to herein as the "R" group. The R group may be substituted or unsubstituted. Two "R" groups "together" form When a compound is written as follows, the R groups and the atoms to which they are attached are cycloalkyl, cycloalkyl, cyclohexyl ... It may form a ring structure such as a chloroalkenyl, aryl, heteroaryl, or heterocycle. It will be understood that the following may be used: for example, but not limited to, NR a R b Group R a and R b When the term "together" is used, they are covalently bonded to each other to form a ring:

[0012] [ka] In addition, two "R" groups form a " When taken together, alternatively, to form a ring, the R groups may be any variable or substituent as defined above. It is not limited to substitution.

[0013] Whenever a group is described as being "optionally substituted," the group is unsubstituted. and may be substituted with one or more of the indicated substituents. Where a group is described as being "unsubstituted or substituted," the When substituted, the substituents may be selected from one or more of the indicated substituents. If no substituents are specified, the indicated "optionally substituted" or or "substituted" groups include alkyl, alkenyl, alkynyl, cycloalkyl ... Alkenyl, acylalkyl, hydroxy, alkoxy, alkoxyalkyl, amino alkyl, amino acid, aryl, heteroaryl, heterocyclyl, aryl(alkyl) , heteroaryl(alkyl), heterocyclyl(alkyl), hydroxyalkyl, Sil, cyano, halogen, thiocarbonyl, O-carbamyl, N-carbamyl, O-thio Carbamyl, N-thiocarbamyl, C-amide, N-amide, S-sulfonamide, N- Sulfonamide, C-carboxy, O-carboxy, isocyanate, thiocyanate, Isothiocyanate, azide, nitro, silyl, sulfenyl, sulfinyl, sulfonyl haloalkyl, haloalkoxy, trihalomethanesulfonyl, trihalomethanesulfon and independently selected from: amide, amino, monosubstituted amino group, and disubstituted amino group. This means that the group may be substituted with one or more groups selected from the group consisting of aryl, ... and aryl.

[0014] As used herein, "C" is a set of integers where "a" and "b" are integers. a ~C b " means the number of carbon atoms in each alkyl, alkenyl or alkynyl group, or cycloalkyl, Intracyclic groups of cycloalkenyl, aryl, heteroaryl, or heteroalicyclyl groups That is, the number of carbon atoms in alkyl, alkenyl, alkynyl, and cycloalkyl ring, cycloalkenyl ring, aryl ring, heteroaryl ring, or heteroalicyclic ring The ring of the alkyl can contain carbon atoms from "a" to "b", inclusive. For example, a "C1-C4 alkyl" group refers to any alkyl group having 1 to 4 carbon atoms. groups, i.e., CH3-, CH3CH2-, CH3CH2CH2-, (CH3)2CH- , CH3CH2CH2CH2-, CH3CH2CH(CH3)- and (CH3)3C- Refers to alkyl, alkenyl, alkynyl, cycloalkyl, cycloalkenyl, aryl, "a" and "b" refer to each aryl, heteroaryl, or heteroalicyclyl group. If no definition is given, the broadest scope set forth in these definitions shall be assumed.

[0015] As used herein, "alkyl" refers to a group that is fully saturated (having both double and triple bonds). The term refers to a linear or branched hydrocarbon chain containing a hydrocarbon group (not including alkyl groups). 0 carbon atoms (wherever it appears herein, it may be "1 to 20" or the like). Any numerical range refers to each integer within the given range; for example, "1 to 20 carbon atoms" , alkyl groups containing 1 carbon atom, 2 carbon atoms, 3 carbon atoms, etc., up to 20 This definition also means that the number of carbon atoms may be less than or equal to 100 carbon atoms, but the ... (This also covers the occurrence of the term "alkyl" when there is no alkyl group.) An alkyl group also has 1 to 10 The alkyl group can also be a medium-sized alkyl having 1 to 6 carbon atoms. It can also be a lower alkyl with carbon atoms. The alkyl group of the compound is "C1-C4 By way of example only, "C1- alkyl" or similar symbols may be used. "C4 alkyl" indicates that the alkyl chain has 1 to 4 carbon atoms, i.e. , alkyl chains are methyl, ethyl, propyl, isopropyl, n-butyl, isobutyl, Typical alkyl groups include those selected from sec-butyl and t-butyl. Examples include, but are not limited to, methyl, ethyl, propyl, isopropyl, butyl, isobutyl butyl, tert-butyl, pentyl, and hexyl. The alkyl groups may be substituted or unsubstituted. It may be substituted or unsubstituted.

[0016] As used herein, "alkenyl" refers to a straight or branched hydrocarbon chain having one or more alkyl groups. It refers to an alkyl group containing one or more double bonds. Examples of alkenyl groups include arethylene and phenyl. Alkenyl groups include vinylmethyl and ethenyl. Alkenyl groups can be unsubstituted or substituted. It may be possible.

[0017] As used herein, "alkynyl" refers to a straight or branched hydrocarbon chain having one or more alkynyl groups. It refers to an alkyl group containing one or more triple bonds. Examples of alkynyl include ethynyl and Alkynyl groups include alkynyl, propynyl, and propynyl. Alkynyl groups can be unsubstituted or substituted.

[0018] As used herein, "cycloalkyl" refers to a fully saturated (both double and triple bonds) alkyl group. refers to a monocyclic or polycyclic hydrocarbon ring system (with or without a ring structure) consisting of two or more rings. When used herein, the rings may be joined in a fused, bridged, or spiro fashion. When used, the term "fused" refers to two groups that have two atoms and one bond in common. As used herein, the term "bridged cycloalkyl" refers to a ring Compounds in which the alkyl contains a linkage of one or more atoms connecting non-adjacent atoms As used herein, the term "spiro" refers to groups having one atom in common. A cycloalkyl group refers to two rings that are not connected by a bridge. has 3 to 30 atoms in the ring, 3 to 20 atoms in the ring, 3 to 10 atoms in the ring, Cycloalkyl can contain 3 to 8 atoms in the ring or 3 to 6 atoms in the ring. The group can be unsubstituted or substituted. Typical monocycloalkyl groups include These include, but are not limited to, cyclopropyl, cyclobutyl, cyclopentyl, These include cyclohexyl, cycloheptyl, and cyclooctyl. Examples of the aryl group are decahydronaphthalenyl, dodecahydro-1H-phenalenyl and tetrahydronaphthalenyl. decahydroanthracenyl; an example of a bridged cycloalkyl group is bicyclo[1.1.1 ]pentyl, bicyclo[2.1.1]heptane, adamantanyl, and norbornanyl Examples of spirocycloalkyl groups include spiro[3.3]heptane and spiro[4 .5] Includes decane.

[0019] As used herein, "cycloalkenyl" refers to a group having at least one alkyl group in one ring. refers to a monocyclic or polycyclic hydrocarbon ring system containing two or more double bonds; If the double bond is above, the π-electron system is completely delocalized throughout the entire ring. cannot be formed (otherwise the group is "aryl" as defined herein). Cycloalkenyl groups contain 3 to 10 atoms in the ring, or 3 to 8 atoms in the ring. When it is composed of two or more rings, these rings may be joined together in a fused manner. The cycloalkenyl group may be unsubstituted or substituted. stomach.

[0020] As used herein, "aryl" refers to a group that is completely delocalized throughout all rings. Carbocyclic (all carbon), monocyclic or polycyclic aromatic rings with localized π electron systems refers to aromatic ring systems (including fused ring systems in which two carbocyclic rings share a chemical bond). The number of carbon atoms in the aryl group can vary. For example, an aryl group can have C6 to C 14 Aryl Even if it is a group, C6~C 10 It may be an aryl group or a C6 aryl group. Examples of aryl groups include, but are not limited to, benzene, naphthalene, and azulene. Aryl groups can be substituted or unsubstituted.

[0021] As used herein, "heteroaryl" refers to one, two, three or more heteroaryls. Atoms other than carbon, i.e., including but not limited to, nitrogen, oxygen, and sulfur Monocyclic or polycyclic aromatic ring systems containing elements (rings with completely delocalized π-electron systems) Heteroaryl groups can have a variety of ring atoms. The alkyl group may have 4 to 14 atoms in the ring, 5 to 10 atoms in the ring, or 5 to 6 atoms in the ring. Additionally, the term "heteroaryl" includes fused ring systems. In this case, the two rings may be, for example, at least one aryl ring and at least one at least one heteroaryl ring, or at least two heteroaryl rings, Examples of heteroaryl rings include, but are not limited to, those described herein. These include those described in the specification and the following: furans, furazans, thiophenes, benzothiophenes Phenol, phthalazine, pyrrole, oxazole, benzoxazole, 1,2,3-oxazole Diazole, 1,2,4-oxadiazole, thiazole, 1,2,3-thiadiazole , 1,2,4-thiadiazole, benzothiazole, imidazole, benzimidazole , indole, indazole, pyrazole, benzopyrazole, isoxazole, benzo Diisoxazole, isothiazole, triazole, benzotriazole, thiadiazo tetrazole, pyridine, pyridazine, pyrimidine, pyrazine, purine, pteridin Quinoline, isoquinoline, quinazoline, quinoxaline, cinnoline, and triazine Heteroaryl groups can be substituted or unsubstituted.

[0022] As used herein, "heterocyclyl" or "heteroalicyclyl" means 3-, 4-, 5-, 6-, 7-, 8-, 9-, 10-, and up to 18-membered monocyclic, bicyclic, and and tricyclic ring systems, in which carbon atoms together with 1 to 5 heteroatoms form the ring system The heterocycle may optionally contain one or more unsaturated bonds. However, this dissonance bond is due to the existence of a completely delocalized π-electron system throughout all rings. Heteroatoms include, but are not limited to, oxygen, sulfur, and An element other than carbon, including nitrogen. A heterocycle may contain one or more carbonyl functional groups or or thiocarbonyl functional groups, so that the definition does not include amides, lactones, cyclic imides, cyclic thioimides and cyclic carbamates This includes all oxo and thio systems. These rings may be fused or spun together as described herein for "cycloalkyl." Additionally, any nitrogen in the heterocyclyl may be quaternized. A heterocyclyl or heteroalicyclyl group may be unsubstituted or Such "heterocyclyl" or "heteroalicyclyl" groups may also be substituted. Examples of include, but are not limited to, those described herein and the following: 1, 3-dioxine, 1,3-dioxane, 1,4-dioxane, 1,2-dioxolane, 1 ,3-dioxolane, 1,4-dioxolane, 1,3-oxathiane, 1,4-oxathiane 1,3,4-oxadiazol-2(3H)-one, 1,2,3-oxadiazoline 1,3-oxathiolane, 1,3-dithiol, 1,3-dithiol Ran, 1,4-oxathiane, tetrahydro-1,4-thiazine, 1,3-thiazinan, 2H-1,2-oxazine, maleimide, succinimide, barbituric acid, thiobarbituric acid uric acid, dioxopiperazine, hydantoin, dihydrouracil, trioxane, hexyl sahydro-1,3,5-triazine, imidazoline, imidazolidine, isoxazoline, Isoxazolidine, oxazoline, oxazolidine, oxazolidinone, thiazoline, thiazoline Azolidine, morpholine, oxirane, piperidine N-oxide, piperidine, piperazine pyrrolidine, pyrrolidone, pyrrolidione, 4-piperidone, pyrazoline, pyrazolidine 2-oxopyrrolidine, tetrahydropyran, 4H-pyran, tetrahydrothiopyran Thiamorpholine, thiamorpholine sulfoxide, thiamorpholine sulfone, and These benzo-fused analogs (e.g., benzimidazolidinone, tetrahydroquinoline, and 3,4-methylenedioxyphenyl).

[0023] As used herein, "aralkyl" and "aryl(alkyl)" refer to a lower alkyl group. It refers to an aryl group connected as a substituent through a lower alkylene group. The alkylene and aryl groups can be substituted or unsubstituted. Examples include, but are not limited to, benzyl, 2-phenylalkyl, 3-phenylalkyl and Naphthyl alkyl is included.

[0024] As used herein, "heteroaralkyl" and "heteroaryl(alkyl)" are )" refers to a heteroaryl group connected as a substituent via a lower alkylene group The lower alkylene and heteroaryl groups of a heteroaralkyl may be substituted or unsubstituted. Examples include, but are not limited to, 2-thienylalkyl, 3-thienylalkyl, enylalkyl, furylalkyl, thienylalkyl, pyrrolylalkyl, pyridylalkyl alkyl, isoxazolylalkyl, imidazolylalkyl and their benzo-fused analogs This includes

[0025] "Heteroalicyclyl(alkyl)" and "heterocyclyl(alkyl)" refer to lower A heterocyclyl group or heteroaryl group connected as a substituent via a cyclic alkylene group Heteroalicyclyl(alkyl) refers to the lower alkylene and heterocyclyl groups. The alkyl group may be substituted or unsubstituted. Examples include, but are not limited to, tetrakis, tetrachloroisothiazolinone ... Hydro-2H-pyran-4-yl(methyl), piperidin-4-yl(ethyl), piperidin Lysin-4-yl(propyl), tetrahydro-2H-thiopyran-4-yl(methyl) and 1,3-thiazin-4-yl(methyl).

[0026] A "lower alkylene group" is a straight-chain -CH2- tethered group that is broken down through its terminal carbon atoms. Forms bonds connecting the parent fragments. Examples include, but are not limited to, methylene (-CH 2-), ethylene (-CH2CH2-), propylene (-CH2CH2CH2-), and and butylene (-CH2CH2CH2CH2-). A lower alkylene group is a lower alkylene group. One or more hydrogens on the alkylene group may be replaced with a substituent listed under the definition of "substituted." Substitution can be achieved by substitution.

[0027] As used herein, "alkoxy" refers to a group of formula -OR, where R is Alkyl, alkenyl, alkynyl, cycloalkyl, cycloalkene, as defined herein. Nyl, aryl, heteroaryl, heterocyclyl, cycloalkyl(alkyl), ari aryl(alkyl), heteroaryl(alkyl), or heterocyclyl(alkyl) A non-limiting list of alkoxy includes methoxy, ethoxy, n-propoxy, 1- Methylethoxy (isopropoxy), cyclopropoxy, n-butoxy, isobutoxy, sec-butoxy, tert-butoxy, cyclobutoxy, phenoxy and benzoxy The alkoxy may be substituted or unsubstituted.

[0028] As used herein, "acyl" refers to an acyl group attached as a substituent via a carbonyl group. The following are hydrogen, alkyl, alkenyl, alkynyl, cycloalkyl, cycloalkenyl, aryl, aryl, heteroaryl, heterocyclyl, cycloalkyl(alkyl), aryl refers to aryl(alkyl), heteroaryl(alkyl), or heterocyclyl(alkyl) Examples include formyl, acetyl, propanoyl, benzoyl and acryl. The acyl may be substituted or unsubstituted.

[0029] As used herein, "acylalkyl" refers to a group that is substituted via a lower alkylene group. Refers to an acyl group attached as a substituent. Examples include aryl-C(=O)-(CH2) n -oh and heteroaryl-C(=O)-(CH2) n - (wherein n is in the range of 1 to 6) is an integer).

[0030] As used herein, "alkoxyalkyl" refers to a group that is substituted through a lower alkylene group. Refers to an alkoxy group attached as a substituent. Examples include C 1~4 Alkyl-O-(CH2 ) n - (wherein n is an integer ranging from 1 to 6).

[0031] As used herein, "aminoalkyl" refers to an amino group substituted via a lower alkylene group. refers to an optionally substituted amino group attached as a group. Examples include H2N(CH2) n - (wherein n is an integer ranging from 1 to 6).

[0032] As used herein, "hydroxyalkyl" refers to a group consisting of one or more hydrogen atoms. refers to an alkyl group that is multiply replaced by a hydroxy group. Dialkyl groups include, but are not limited to, 2-hydroxyethyl, 3-hydroxypropyl, 4-hydroxypropyl, 5-hydroxypropyl, 6-hydroxypropyl, 7-hydroxypropyl, 8-hydroxypropyl, 9-hydroxypropyl, 10-hydroxypropyl, 11-hydroxypropyl, 12-hydroxypropyl, 13-hydroxypropyl, 14-hydroxypropyl, 15-hydroxypropyl, 16-hydroxypropyl, 17-hydroxypropyl, 18-hydroxypropyl, 19-hydroxy Hydroxypropyl, 2-hydroxypropyl, and 2,2-dihydroxyethyl. The oxyalkyl may be substituted or unsubstituted.

[0033] As used herein, "haloalkyl" refers to one or more hydrogen atoms. is replaced by a halogen (e.g., monohaloalkyl, dihaloalkyl, Such groups include, but are not limited to, chlorine, chlorine- ... Chloromethyl, fluoromethyl, difluoromethyl, trifluoromethyl, chloro-fluoro chloro-difluoroalkyl and 2-fluoroisobutyl. The alkyl may be substituted or unsubstituted.

[0034] As used herein, "haloalkoxy" refers to an alkoxy group that is substituted with one or more hydrogen atoms. Alkoxy groups in which a number of groups are replaced by halogen (e.g., monohaloalkoxy, Such groups include, but are not limited to, dihaloalkoxy and trihaloalkoxy. However, chloromethoxy, fluoromethoxy, difluoromethoxy, trifluoromethoxy chloro-fluoroalkyl, chloro-difluoroalkoxy and 2-fluoroiso Haloalkoxy may be substituted or unsubstituted, including butoxy.

[0035] A "sulfenyl" group refers to a "-SR" group, where R is hydrogen, alkyl, alkoxy, or methyl. aryl, alkynyl, cycloalkyl, cycloalkenyl, aryl, heteroaryl, Tetracyclyl, cycloalkyl(alkyl), aryl(alkyl), heteroaryl( It can be either heterocyclyl(alkyl) or heterocyclyl(alkyl). It may be substituted or unsubstituted.

[0036] A "sulfinyl" group refers to a "-S(=O)-R" group, where R is sulfenyl (The term "sulfinyl" can be used interchangeably with "sulfinyl" and "sulfinyl"). It may be unsubstituted.

[0037] A "sulfonyl" group refers to an "SO2R" group, where R is as defined for sulfenyl. (The term "sulfonyl" can be used interchangeably with "substituted or unsubstituted"). That's fine.

[0038] An "O-carboxy" group refers to an "RC(=O)O-" group, where R is as defined herein. As defined above, hydrogen, alkyl, alkenyl, alkynyl, cycloalkyl, cycloalkenyl , aryl, heteroaryl, heterocyclyl, cycloalkyl (alkyl), aryl (alkyl), heteroaryl(alkyl) or heterocyclyl(alkyl). The O-carboxy may be substituted or unsubstituted.

[0039] The terms "ester" and "C-carboxy" refer to the group "-C(=O)OR" (wherein R can be the same as defined for O-carboxy). The ester and C-carboxy may be substituted or unsubstituted.

[0040] A "thiocarbonyl" group refers to a "-C(=S)R" group, where R is O-carboxy. (The definition for thiocarbonyl may be the same as that for thiocarbonyl.) It may be substituted or unsubstituted.

[0041] A "trihalomethanesulfonyl" group refers to an "X3CSO2-" group, where each X is a halomethanesulfonyl group. Rogen).

[0042] The "trihalomethanesulfonamide" group is "X3CS(O)2N(RA )-" refers to the group where each X is a halogen and R A is hydrogen, alkyl, alkenyl, alkynyl, Cycloalkyl, cycloalkenyl, aryl, heteroaryl, heterocyclyl, cyclo alkyl(alkyl), aryl(alkyl), heteroaryl(alkyl) or heteroaryl(alkyl) It is a terecyclyl(alkyl).

[0043] As used herein, the term "amino" refers to the group --NH.sub.2.

[0044] As used herein, the term "hydroxy" refers to an --OH group.

[0045] A "cyano" group refers to a "-CN" group.

[0046] As used herein, the term "azido" refers to the group --N3.

[0047] An "isocyanate" group refers to a "-NCO" group.

[0048] A "thiocyanate" group refers to a "-CNS" group.

[0049] An "isothiocyanate" group refers to a "-NCS" group.

[0050] A "carbonyl" group refers to a C=O group.

[0051] The "S-sulfonamide" group is "-SO2N(R A R B ) group, where R A oh Yobi R B are independently hydrogen, alkyl, alkenyl, alkynyl, cycloalkyl, cycloalkyl Alkenyl, aryl, heteroaryl, heterocyclyl, cycloalkyl (alkyl ), aryl(alkyl), heteroaryl(alkyl) or heterocyclyl(alkyl) S-sulfonamides can be substituted or unsubstituted. good.

[0052] The "N-sulfonamide group" is "RSO2N(R A )— group, where R and R A are independently hydrogen, alkyl, alkenyl, alkynyl, cycloalkyl, cycloa alkenyl, aryl, heteroaryl, heterocyclyl, cycloalkyl(alkyl), Aryl(alkyl), heteroaryl(alkyl), or heterocyclyl(alkyl) The N-sulfonamide may be substituted or unsubstituted. .

[0053] The "O-carbamyl" group is defined as "-OC(=O)N(R A R B ) group, where R A and R B are independently hydrogen, alkyl, alkenyl, alkynyl, cycloalkyl, cyclohexyl, cycloalkenyl, aryl, heteroaryl, heterocyclyl, cycloalkyl (alkoxy) aryl), aryl(alkyl), heteroaryl(alkyl) or heterocyclyl(aryl) O-carbamyl may be substituted or unsubstituted. stomach.

[0054] The "N-carbamyl" group is "ROC(=O)N(R A (wherein R and BiR A are independently hydrogen, alkyl, alkenyl, alkynyl, cycloalkyl, cyclo Alkenyl, aryl, heteroaryl, heterocyclyl, cycloalkyl(alkyl) , aryl(alkyl), heteroaryl(alkyl) or heterocyclyl(alkyl) )). N-carbamyl can be substituted or unsubstituted.

[0055] The "O-thiocarbamyl" group is defined as "-OC(=S)-N(R A R B ) group (wherein , R A and R B are independently hydrogen, alkyl, alkenyl, alkynyl, cycloalkoxy aryl, cycloalkenyl, aryl, heteroaryl, heterocyclyl, cycloalkyl ( alkyl), aryl(alkyl), heteroaryl(alkyl) or heterocyclyl O-Thiocarbamyl can be substituted or unsubstituted. It's okay to have one.

[0056] The "N-thiocarbamyl group" is "ROC(=S)N(R A )— group, where R and R A are independently hydrogen, alkyl, alkenyl, alkynyl, cycloalkyl, cyclohexyl, cycloalkenyl, aryl, heteroaryl, heterocyclyl, cycloalkyl (alkoxy) aryl), aryl(alkyl), heteroaryl(alkyl) or heterocyclyl(aryl) N-thiocarbamyl can be substituted or unsubstituted. Good too.

[0057] A "C-amide" group is a group consisting of -C(=O)N(R A R B ) group, where R A and R B are independently hydrogen, alkyl, alkenyl, alkynyl, cycloalkyl, cycloa alkenyl, aryl, heteroaryl, heterocyclyl, cycloalkyl(alkyl), Aryl(alkyl), heteroaryl(alkyl), or heterocyclyl(alkyl) The C-amide may be substituted or unsubstituted.

[0058] The "N-amide" group is defined as "RC(=O)N(R A )— group, where R and R A are independently hydrogen, alkyl, alkenyl, alkynyl, cycloalkyl, cycloalkene Nyl, aryl, heteroaryl, heterocyclyl, cycloalkyl(alkyl), ari aryl(alkyl), heteroaryl(alkyl) or heterocyclyl(alkyl). The N-amide may be substituted or unsubstituted.

[0059] The "urea" group is defined as "N(R)-C(=O)-NR A R B " group (wherein R is hydrogen or or alkyl, and R A and R B are independently hydrogen, alkyl, or alkenyl , alkynyl, cycloalkyl, cycloalkenyl, aryl, heteroaryl, hetero Cyclyl, cycloalkyl(alkyl), aryl(alkyl), heteroaryl(aryl) The urea may be substituted or unsubstituted. It may be substituted or unsubstituted.

[0060] The "oxime" group is "-C(=N-OH)R A " (wherein R A is independent and alkyl, alkenyl, alkynyl, cycloalkyl, cycloalkenyl, aryl, hetero aryl, heterocyclyl, cycloalkyl(alkyl), aryl(alkyl), hetero It can be heteroaryl(alkyl) or heterocyclyl(alkyl). The oxime may be substituted or unsubstituted.

[0061] "Acylhydrozone" means "-C(=N-NH-acyl)-R A .” (wherein, The acyl moiety has the structure provided herein for "acyl," R A is independent, Alkyl, alkenyl, alkynyl, cycloalkyl, cycloalkenyl, aryl, hexane Heteroaryl, heterocyclyl, cycloalkyl(alkyl), aryl(alkyl), It can be heteroaryl(alkyl) or heterocyclyl(alkyl). The acylhydrozones may be substituted or unsubstituted.

[0062] "Hydrazine" is "-NHNR A R B " (wherein R A and R B is independent Hydrogen, alkyl, alkenyl, alkynyl, cycloalkyl, cycloalkenyl, aryl aryl, heteroaryl, heterocyclyl, cycloalkyl(alkyl), aryl(alkyl) heteroaryl(alkyl), heteroaryl(alkyl), or heterocyclyl(alkyl) The hydrazine may be substituted or unsubstituted.

[0063] As used herein, the term "halogen atom" or "halogen" refers to fluorine, chlor ... means any one of the radiostable atoms in column 7 of the periodic table, such as fluorine, bromine, and iodine. do.

[0064] As used herein,

[0065] [ka] indicates a single or double bond unless otherwise indicated.

[0066] If the number of substituents is not specified (e.g., haloalkyl), one or more substituents are allowed. For example, "haloalkyl" means a group consisting of the same or different halogens. It may contain one or more of the following. Another example is "C1-C3 alkoxyphenyl "Nyl" refers to the same or different alkoxy groups containing 1, 2, or 3 atoms. It may contain one or more of them.

[0067] As used herein, abbreviations for all protecting groups, amino acids and other compounds are , unless otherwise indicated, their common usage, accepted abbreviations, or biochemical nomenclature to the IUPAC-IUB Commission on the Law of Chemical Substances (see Biochem. 11:942-944 (1972)). follow.

[0068] As used herein, "protecting group" and "protecting groups" (and abbreviation "PG") refer to The term "protected" refers to a group present in a molecule that is protected from undesired chemical reactions. Protecting group refers to any atom or group of atoms that is added to a molecule to protect it from the chemical reaction. Examples of protecting group moieties are TW Greene and PGM Wuts, Protective Groups in Organic Synthesis, 3. Ed. John Wi ley & Sons, 1999, and in JFW McOmie, Protective Groups in Organic Chemistry and JP 2002-102263, both of which disclose suitable protecting groups. The protecting group moiety is a group in which the protecting group is It is stable to certain reaction conditions and can be prepared using methodologies known in the art. The protecting groups can be selected so as to be easily removed at a convenient stage using a suitable protecting group. A typical list includes: benzyl; substituted benzyl; alkylcarbonyl and Alkoxycarbonyl (e.g., t-butoxycarbonyl (BOC), acetyl, or isobutyryl; arylalkylcarbonyl and arylalkoxycarbonyl (e.g. benzyloxycarbonyl); substituted methyl ethers (e.g., methoxymethyl ether) substituted ethyl ethers; substituted benzyl ethers; tetrahydropyranyl ethers; silyl (e.g., trimethylsilyl, triethylsilyl, triisopropylsilyl, t-butylsilyl) ethyldimethylsilyl, tri-isopropylsilyloxymethyl, [2-(trimethylsilyl) methyl or t-butyldiphenylsilyl); esters (e.g., benzoic acid Acid esters; carbonates (e.g., methoxymethyl carbonate) xymethylcarbonate); sulfonate (e.g., tosylate or or mesylate; acyclic ketals (e.g., dimethyl acetal); cyclic ketals Tars (e.g., 1,3-dioxane, 1,3-dioxolane, and the like described herein) acyclic acetals; cyclic acetals (e.g., those described herein); acyclic Hemiacetals; cyclic hemiacetals; cyclic dithioketals (e.g., 1,3-dithiane or 1,3-dithiolane); orthoesters (e.g., those described herein); triarylmethyl groups (e.g., trityl; monomethoxytrityl (MMTr); 4, 4'-Dimethoxytrityl (DMTr); 4,4',4"-Trimethoxytrityl (TM Tr); and those described herein).

[0069] As used herein, the term "leaving group" (and the abbreviation "LG") refers to a group that is a group that is released in a chemical reaction. refers to any atom or moiety that can be replaced by another atom or moiety. More specifically, in some embodiments, a "leaving group" refers to a group that is replaced in a nucleophilic substitution reaction. In some embodiments, a "leaving group" refers to an atom or moiety that is a conjugate base of a strong acid. Examples of suitable leaving groups include, but are not limited to, thiazolinone ... Sylate, mesylate, trifluoroacetate Non-limiting examples of leaving groups include aryl, aryl cetate, and halogens (e.g., I, Br, and Cl). For typical features and examples, see, e.g., Organic Chemistry, 2nd ed., Francis Carey (1999). 92), pages 328-331;Introduction to Organic Chemistry, 2d ed., Andrew Streitwies er and Clayton Heathcock (1981), pages 169-171; and Organic Chemistry, 5th ed. , John McMurry (2000), pages 398 and 408; all of these , incorporated herein by reference for the limited purpose of disclosing characteristics and examples of leaving groups. will be incorporated into

[0070] The term "pharmaceutically acceptable salt" means a compound that does not cause significant irritation to an organism to which it is administered. and do not neutralize the biological activity and properties of the compound. In some embodiments, the salt is an acid addition salt of the compound. Pharmaceutical salts are salts of the compound. The mixture is treated with a hydrohalic acid (e.g., hydrochloric or hydrobromic acid), sulfuric acid, nitric acid, and phosphoric acid. Pharmaceutical salts can be obtained by reacting a compound with a fatty acid. organic acids such as aromatic or aromatic carboxylic or sulfonic acids, for example, formic acid, acetic acid, Citric acid, lactic acid, malic acid, tartaric acid, citric acid, ascorbic acid, nicotinic acid, methanesulfonate sulfonic acid, ethanesulfonic acid, p-toluenesulfonic acid, salicylic acid or naphthalene sulfonic acid Pharmaceutical salts can also be obtained by reacting a compound with a base. to produce a salt such as an ammonium salt, an alkali salt such as a sodium salt or a potassium salt. Metal salts, alkaline earth metal salts such as calcium salts or magnesium salts, dicyclohexyl Aminomethylamine, N-methyl-D-glucamine, tris(hydroxymethyl)methylamine, C 1~C7 alkylamine, cyclohexylamine, triethanolamine, ethylenediamine It forms salts with organic bases such as amine, as well as with amino acids such as arginine and lysine. It can also be obtained by

[0071] Terms and phrases used in this application and variations thereof are particularly intended to be illustrative and not restrictive of the principles of the present invention. In the claims, unless expressly stated, the scope of the invention is open-ended as opposed to limiting. In the example above, "includes (including, The term "including" means "including, but not limited to..." including, without limitation) and including but not limited to )" and the like; as used herein, "including (com The term "including," "containing," "containing" or "characterized by" is synonymous with "open-ended" and is inclusive or open-ended; "Having" does not exclude additional, unrecited elements or method steps. The term "having at least" is to be interpreted as "having at least" should; the term "includes" means "they Includes but is not limited to The term "examples" should be interpreted as "exhaustive" and "limited to"; used to provide illustrative examples of the matters under discussion rather than as a comprehensive or exclusive enumeration; "preferably," "preferred," "desired" Terms such as "desired" or "desirable" and The use of terms such as "structure" and "function" indicates that certain characteristics are critical to structure or function. should not be understood to imply that it is essential or even important, Instead, alternative or additional features may be used that may or may not be utilized in a particular embodiment. It should be understood that the word "including" is merely intended to emphasize the characteristics. The term "comprising" is used to mean "having at least "including at least" or "including at least" When used in the context of a method, the phrase " The term "comprising" means that the method comprises at least the recited steps. It means that the compound, composition or When used in the context of a device, the term "comprising" is that the compound, composition or device contains at least the described properties or components. , which means that the term may also include additional properties or components. A group of items connected by "call" indicates that each such item is present in that grouping. rather, unless the context indicates otherwise, should be read as "and / or." Similarly, a group connected by the conjunction "or" The clause should not be read to require mutual exclusivity within the group. Instead, it should be read as "and / or" unless the context dictates otherwise. do.

[0072] Regarding the use of substantially any plural and / or singular terminology herein , those skilled in the art will readily appreciate the possibility of converting from plural to singular, as appropriate to the context and / or application. and / or from singular to plural. Substitutions may be explicitly indicated in this specification for clarity. "An" does not exclude pluralism. The mere fact that a combination of these measures is listed in the program does not necessarily mean that it can be used to advantage. Any reference signs in the claims do not indicate that the invention cannot be , should not be construed as limiting the scope.

[0073] In any compound described herein that has one or more chiral centers, the absolute configuration If the configuration chemistry is not explicitly indicated, each center may independently be in the R-configuration or It is understood that can be in the S-configuration or a mixture thereof. The compounds provided herein may be enantiomerically pure or enantiomerically enriched. Whether as a racemic mixture or diastereoisomerically pure, It may be either stereoisomerically enriched or a mixture of stereoisomers. The present specification also provides a method for preparing a compound having one or more double bonds that produce geometric isomers that can be defined. In any compound described herein, each double bond may independently be E or Z. It is understood that the present invention may be any of a variety of materials, including but not limited to, fluororesin, fluoroisotope ...

[0074] Likewise, all tautomeric forms of any compound described are intended to be included. It is understood that

[0075] When a compound disclosed herein has an open valence, the valence may be replaced by hydrogen or its equivalent. By filling it with hydrogen atoms, e.g., hydrogen-1 (protium) and hydrogen-2 (deuterium) I want you to understand that this will happen.

[0076] It is understood that the compounds described herein can be isotopically labeled. Substitution with any isotope may result in, for example, an increase in in vivo half-life or a decrease in required dosage. may confer certain therapeutic advantages resulting from greater metabolic stability, such as Each chemical element represented in a compound structure can include any isotope of that element. For example, in a compound structure, hydrogen atoms may be explicitly disclosed as being present in the compound. It may be or may be understood to be the case that hydrogen atoms are present. At any position of the compound, the hydrogen atom can be, but is not limited to, hydrogen Any isotope of hydrogen, including hydrogen-1 (protium) and hydrogen-2 (deuterium) Therefore, references herein to compounds are possible unless the context dictates otherwise. The term "isotopic forms" encompasses all isotopic forms in which

[0077] The methods and combinations described herein include those containing crystalline forms (also known as polymorphs, which are compounds that are produced in the same manner as the compound). (including different crystal packing arrangements of compounds of the same elemental composition), amorphous phases, salts, solvates, and It is understood that the compounds described herein include hydrates and hydrates. exists in solvated form with pharmaceutically acceptable solvents such as water, ethanol, etc. In embodiments, the compounds described herein exist in unsolvated form. Solvates are chemically It contains either stoichiometric or non-stoichiometric amounts of solvents, such as water, ethanol, etc. It can be formed during the process of crystallization using pharmaceutically acceptable solvents such as Hydrates are formed when the solvent is water, or alcohols when the solvent is alcohol. In addition, the compounds provided herein may be used in unsolvated as well as solvated forms. In general, solvated forms are available for the compounds provided herein and For purposes of the present invention and methods, the unsolvated forms are considered equivalent.

[0078] Where a range of values ​​is provided, the upper and lower limits, as well as any intervening values ​​between the upper and lower limits of the range, It is understood that each of the values ​​is encompassed within the present embodiment.

[0079] As used herein, "RNA" refers to naturally occurring or non-naturally occurring It refers to ribonucleic acid. For example, RNA is a molecule made up of one or more nucleic acid bases, nucleosides, nucleobases, and nucleotides. may contain modified and / or non-naturally occurring components such as nucleotides, nucleotides, or linkers. RNA contains a cap structure, chain-terminating nucleosides, stem-loops, and poly(A) sequences. The RNA may contain a polyadenylation signal, a nucleotide sequence, and / or a polyadenylation signal. For example, RNA can have a nucleotide sequence that encodes a message peptide. mRNA that encodes a specific polypeptide. Translation of the mRNA, e.g., in vivo translation of the mRNA inside a mammalian cell, results in the production of the encoded polypeptide. RNA can be used to produce small interfering RNA (siRNA), miRNA, and croRNA (miRNA), Dicer substrate RNA (dsRNA), small hairpin RNA ( shRNA), mRNA, single guide RNA (sgRNA), cas9 mRNA, and mixtures thereof.

[0080] The terms "polypeptide," "peptide," and "protein" refer to a group of peptide bonds. Used interchangeably to refer to a string of at least three amino acids linked together by The term "peptide" can refer to an individual peptide or a collection of peptides. Peptides may contain natural amino acids, unnatural amino acids (i.e., amino acids not found in nature), compounds that are not present in the polypeptide chain but can be incorporated into the polypeptide chain), and / or amino acid analytes Also, one or more of the amino acids in the peptide may be For example, carbohydrate groups, phosphate groups, farnesyl groups, isofarnesyl groups, fatty acid groups, Addition of chemical entities such as linkers for jugation, functionalization, or other modifications Modifications can be made by cyclization of the peptide, incorporation of D-amino acids, etc. may be included.

[0081] As used herein, "treat," "treating," "treatment," and "therapy" The term "use above" refers to the elimination, reduction, or treatment of one or more symptoms of a disease or disorder. As used herein, a "therapeutically effective amount" refers to an improvement in the clinical significance of such symptoms. An effect refers to an amount of a therapeutic agent sufficient to mediate some elimination, reduction, or amelioration of a condition. If the magnitude is sufficient to affect the health or prognosis of the recipient subject, clinical A therapeutically effective amount is an amount that delays or minimizes the onset of disease, e.g., Can refer to an amount of a therapeutic drug sufficient to slow or minimize the spread of cancer A therapeutically effective amount also refers to the amount of a therapeutic agent that provides a therapeutic benefit in the treatment or management of a disease. You can point to it.

[0082] The compositions described herein are preferably provided in unit dosage form. When used in a "unit dosage form," a "unit dosage form" refers to a dosage form administered to an animal, preferably a mammalian subject, in accordance with good medical practice. The composition contains an amount of the compound or composition suitable for administration in a single dose. However, the preparation of single or unit dosage forms is intended to provide a single dose once daily or once per course of treatment. It does not imply that a single dose will be administered. A single dose is not specifically excluded. However, such dosage forms are intended to be administered once, twice, three or more times per day. It is administered as an infusion over a period of time (e.g., from 30 minutes to about 2-6 hours). It may be administered as a continuous infusion or given multiple times over the course of treatment. Those skilled in the art will appreciate that the formulation does not specifically contemplate an entire course of treatment, but rather It is recognized that such decisions are left to those skilled in the art of treatment rather than formulation. As used herein, the term "prophylactic" refers to a drug or a method for treating a disorder or disease. It can be used to prevent such disorders or diseases before any symptoms are detected. A "prophylactically effective" amount is an amount of a prophylactic agent sufficient to mediate such prevention. A prophylactically effective amount also refers to the amount of a prophylactic agent that provides a prophylactic benefit in the prevention of disease. You can also point.

[0083] The useful compositions described above can be used in a variety of applications, including for administration, e.g., orally, nasally, rectally, topically (including transdermally), or intravenously. ), ophthalmic, intracerebral, intracranial, intrathecal, intraarterial, intravenous, intramuscular, or other parenteral routes The composition may be in any of a variety of forms suitable for a variety of routes of administration. Oral and nasal compositions may be administered by inhalation, using available methodologies. It will be understood that the present invention includes compositions made using the specific dosage form desired. Depending on the route, various pharmaceutically acceptable carriers well known in the art may be used. Pharmaceutically acceptable carriers include, for example, solid or liquid fillers, diluents, and the like. agents, hydrotropes, surfactants, and encapsulating materials. Optionally, a pharmaceutically active material may be included that does not substantially interfere with the inhibitory activity of the compound. The amount of carrier used in combination with the compound may be 100 mg / kg or more per unit dose of the compound. The method described herein is sufficient to provide a practical amount of material for administration. Techniques and compositions for preparing dosage forms useful in this field are described in the following references: Modern Pharmaceutics, 4th Ed., Cha pters 9 and 10 (Banker & Rhodes, editors, 2002);Lieberman et al., Pharmaceutica Dosage Forms: Tablets (1989); and Ansel, Introduction to Pharmaceutical Dosage ge Forms 8th Edition (2004).

[0084] As used herein, the terms "individual," "host," "subject," and "patient" are used interchangeably. The terms are used interchangeably herein and refer to animals such as, but not limited to, humans, mice and rats. Refers to any mammal, including rodents and other laboratory animals.

[0085] As used herein, the term "pharmaceutically acceptable carrier" refers to a phosphate buffered saline solution. Saline solutions, emulsions such as water and oil / water or water / oil emulsions, and seeds The compositions may include any of the standard pharmaceutical carriers, such as various types of wetting agents.

[0086] The term "PEG-lipid" refers to a lipid modified with polyethylene glycol. Exemplary PEG-lipids include, but are not limited to, C 14 PEG 350 , C 14 PEG 1000 , C 14 PEG 2000 , C 14 PEG 3000 , and C 18 PEG 2000 Includes:

[0087] The term "oligonucleotide" refers to a short DNA fragment containing a relatively small number of nucleotides. A refers to a molecule or oligomer of RNA, DNA, or RNA.

[0088] A. Lipid Nanoparticles Effective targeted delivery of bioactive agents such as small molecule drugs, proteins, and nucleic acids is a key component of medicine. Nucleic acid delivery is a continuing challenge in the field, particularly due to the relative instability of nucleic acids and the cellular This is complicated by the low cell permeability. Lipid nanoparticles containing lipids can more effectively deliver nucleic acids to specific tissues in the body. In one embodiment, the lipid nanoparticles comprise a nucleic acid that is encapsulated in an ionizable lipid. By mixing with PEG-lipids, phospholipids, cholesterol, and optionally nucleic acids, In some embodiments, the lipid nanoparticles can be formulated to target In some embodiments, the disclosed lipid nanoparticles do not contain a targeting ligand. Preferential targeting of T cells over hepatocytes in the absence of gliomas.

[0089] The size of the lipid nanoparticles varies. In one embodiment, the lipid nanoparticles range from about 30 to about 17 The lipid nanoparticles can have an average hydrodynamic diameter of between about 30 nm and about 40 nm. , 35nm, 40nm, 45nm, 50nm, 55nm, 60nm, 65nm, 70nm , 75nm, 80nm, 85nm, 90nm, 95nm, 100nm, 105nm, 11 0nm, 115nm, 120nm, 125nm, 130nm, 135nm, 140nm, 145nm, 150nm, 155nm, 160nm, 165nm, 170nm, or Any range with endpoints defined by any two of the values ​​listed above. For example, in one embodiment, the nanoparticles may have a uniform hydrodynamic diameter of 50 nm or less. They have an average hydrodynamic diameter ranging from between ∼100 nm.

[0090] 1.Compound Some embodiments described herein include compounds of formula (I):

[0091] [ka] The present invention relates to the compound

[0092] In various embodiments, the compounds of formula (I) may be iodinated, as described elsewhere herein. In various embodiments, the R 1 is C9~C 20 Alkyl or C9-C with 1-3 units of unsaturation 20 For example, some embodiments include In the form, R of formula (I) 1 is the expression

[0093] [ka] C 17 C9-C with two units of unsaturation, such as alkenyl groups 20 It is alkenyl.

[0094] In various embodiments, X in formula (I)1 and X 2 are each independently absent or or -O-, -NR 2 - and

[0095] [ka] where R 2 is hydrogen or C1-C6 alkyl, and a is between 1 and 6. is an integer, and X 7 are independently hydrogen, hydroxyl, or -NR 6 R 7 and R 6 and R 7 are each independently hydrogen or C1-C6 alkyl; or Alternatively, they may be joined together with the nitrogen to which they are attached to form one or two C1-C6 alkyl groups. and forming a 4- to 7-membered heterocyclyl optionally substituted with a methyl group, wherein the heterocyclyl The alkyl optionally contains additional heteroatoms selected from oxygen, sulfur, and nitrogen. In some embodiments, X 1 is absent or X 2 is absent or X 1 and X 2 As described elsewhere herein, X 1 -X 2 -X 3 -X 4 do not contain any oxygen-oxygen, oxygen-nitrogen, or nitrogen-nitrogen bonds between each other. Therefore, X 1 and X 2 cannot both be -O- and both be -NR 2 - Similarly, X 1 and X 2 are -O- and -NR, respectively. 2 -Yes and -NR 2 - and -O-.

[0096] In various embodiments, X 1 is —O—. In various embodiments, X 2 is -O- In some embodiments, X 1 is -(CH2) a -, -CH(OH)-, or -( CH2) a-1 CH(OH)-, etc.

[0097] [ka] In some embodiments, X 2 is -(CH2) a -, -CH(OH)-, or - (CH2) a-1 CH(OH)-, etc.

[0098] [ka] In various embodiments, each a is independently 1, 2, 3, 4, 5, or 6. In various embodiments, X 1 is -NR 6 R 7 In various embodiments, X 2 -N R 6 R 7 In some embodiments, R 6 is hydrogen or C1-C6 alkyl. In some embodiments, R 7 is hydrogen or C1-C6 alkyl. , R 6 and R 7 These, together with the nitrogen to which they are attached, form one or two C1~ Forms a 4- to 7-membered heterocyclyl optionally substituted with a C6 alkyl group. In embodiments, R 6 and R 7 4-7 membered compound formed by combining The heterocyclyl of the formula (I) contains an additional heteroatom selected from oxygen, sulfur, and nitrogen. .

[0099] In various embodiments, X in formula (I) 3 and X 4 are each independently absent or or (1) A 4- to 8-membered heterocyclic group optionally substituted with one or two C1-C6 alkyl groups. Telocyclyl; (2) a 5- to 6-membered heterocyclic group optionally substituted with one or two C1-C6 alkyl groups; Teloaryl; (3) 5-6-membered alkyl groups optionally substituted with one or two C1-C6 alkyl groups. reel; (4) a 4- to 7-membered cyclohexyl group optionally substituted with one or two C1-C6 alkyl groups; Chloroalkyl; (5) -O-; or (6)-NR 3 -(In the formula, each R 3 are independently a hydrogen atom or a C1-C6 alkyl. ( is selected from.

[0100] In some embodiments, X 3 is absent or X 4 is absent or X 3 and X 4 As described elsewhere herein, X 1 -X 2 -X 3 -X 4 do not contain any oxygen-oxygen, oxygen-nitrogen, or nitrogen-nitrogen bonds between each other. Therefore, X 2 and X3 and X cannot both be -O-. 2 -O- or -NR 2 -If X 3 is -NR 3 - should not be the same. Similarly, X 3 -O- or -NR 3 -If X 2 is -NR 2 - should not be. Similarly, X 3 and X 4 cannot both be -O- and both be -NR 3 -It should not be Similarly, X 3 and X 4 are -O- and -NR, respectively. 3 -Should not be , respectively -NR 3 - and -O-.

[0101] In various embodiments, X in formula (I) 3 and X 4 are each independently 1 or 2 4-8 membered heterocyclic alkyl group optionally substituted with C1-C6 or C1-C3 alkyl groups For example, in various embodiments, X 3 and X 4 are each independently azetidinyl, methylazetidinyl, pyrrolidinyl, methylpyrrolidinyl, piperidinyl, Methylpiperidinyl, piperazinyl, methylpiperazinyl, dimethylpiperazinyl, mol Diazepanyl, diazepanyl, methyldiazepanyl, octahydro-2H-quinolizinyl, Azabicyclo[3.2.1]octyl, methyl-azabicyclo[3.2.1]octyl, diazabicyclo[3.2.1]octyl azaspiro[3.5]nonyl, or methyldiazaspiro[3.5]nonyl.

[0102] In various embodiments, X in formula (I) 3 and X 4 are each independently 1 or 2 5-6 membered heteroaryl optionally substituted with C1-C6 or C1-C3 alkyl groups For example, in various embodiments, X 3 and X 4 are each independently pyrrolyl, methylpyrrolyl, imidazolyl, methylimidazolyl, pyridinyl, or methyl It is pyridinyl.

[0103] In various embodiments, X in formula (I) 3 and X 4 are each independently 1 or 2 5-6 membered aryl optionally substituted with C1-C6 or C1-C3 alkyl groups For example, in various embodiments, X 3 and X 4 are each independently phenyl , methylphenyl, naphthyl, or methylnaphthyl.

[0104] In various embodiments, X in formula (I) 3 and X 4 are each independently 1 or 2 4- to 7-membered cycloaliphatic group optionally substituted with C1-C6 or C1-C3 alkyl groups For example, in various embodiments, X 3 and X 4 are each independently cyclopentyl, methylcyclopentyl, cyclohexyl, or methylcyclohexyl be.

[0105] In various embodiments, X in formula (I) 3 is -O-. In another embodiment, X 4 is —O—. In various embodiments, X 3Ha-NR 3 -, where R 3 teeth, It is a hydrogen atom or a C1-C6 alkyl, such as a C1-C3 alkyl. For example, various In embodiments, X 3 is -N(CH3)-, -N(CH2CH3)-, or N(CH2 In another embodiment, X 4 Ha-NR 3 -, where R 3 teeth , a hydrogen atom, or a C1-C6 alkyl such as a C1-C3 alkyl. In some embodiments, X 4 is -N(CH3)-, -N(CH2CH3)-, or N(CH 2CH2CH3)-.

[0106] In various embodiments, X in formula (I) 5 Ha-(CH2) b -, where b is 0~ In some embodiments, b is 0, in which case X 5 is absent In other embodiments, b is 1, 2, 3, 4, 5, or 6.

[0107] In various embodiments, X in formula (I) 6 is hydrogen, C1-C6 alkyl, 1 or 2 5-6 membered heteroaryl optionally substituted with a C1-C6 alkyl group or -N R 4 R 5 In some embodiments, R 4 and R 5 are each independently hydrogen or Alternatively, in other embodiments, R 4 and R 5 These are Together with the attached nitrogen, one or two C1-C6 alkyl groups as needed Form a substituted 4- to 7-membered heterocyclyl, wherein the 4- to 7-membered heterocyclyl is It optionally contains additional heteroatoms selected from oxygen, sulfur, and nitrogen.

[0108] In various embodiments of formula (I), X 1 , X 2 , X 3 , X 4 , and X 5 At least one of For example, in various embodiments, formula (I) includes at least one of X 1 , X 2 , X 3 , X 4 , and X 5 In another embodiment, at least two of the following are present: , X 1 , X 2 , X 3 , X 4 , and X 5 There are at least three of these. In some embodiments, formula (I) includes X 1 , X 2 , X 3 , X 4 , and X 5 At least one of In another embodiment, there are four X 1 , X 2 , X 3 , X 4 , and and X 5 All of these exist.

[0109] In some embodiments, X 6 is hydrogen. In other embodiments, X 6 is C1~C3 al and C1-C6 alkyl, such as methyl, ethyl, or propyl. In an embodiment of the present invention, X 6 is optionally substituted with one or two C1-C6 alkyl groups For example, in various embodiments, X 6 is pyrrolyl, Methylpyrrolyl, imidazolyl, methylimidazolyl, pyridinyl, or methylpyridinyl In another embodiment, X is nyl. 6 Ha-NR 4 R 5 For example, in some embodiments is X 6 are -NH2, -NHCH3, -NHCH2CH3, -NHCH2CH2CH3 , -N(CH3)2, -N(CH2CH3)2, or -N(CH2CH2CH3)2 Alternatively, in other embodiments, R 4 and R 5 are the nitrogen atoms to which they are bonded. Together they form a 4- to 7-membered heterocyclyl. A 4- to 7-membered heterocyclyl is a C optionally substituted with one or two C1-C6 alkyl groups, such as 1-C3 alkyl; and / or the 4- to 7-membered heterocyclyl may be selected from oxygen, sulfur, and nitrogen. For example, in some embodiments, In terms of form, X 6 are azetidinyl, methylazetidinyl, pyrrolidinyl, methylpyrrolidinyl nyl, piperidinyl, methylpiperidinyl, piperazinyl, methylpiperazinyl, dimethy The aryl amine may be rupiperazinyl, morpholinyl, diazepanyl, or methyldiazepanyl.

[0110] In various embodiments, each X in formula (I) 7 is hydrogen. In other embodiments, each X 7 Hahi In another embodiment, each X 7 Ha-NR 6 R 7 a is between 2 and 6 In one embodiment, each X 7may be the same or different. For example, In various embodiments, X 7 is -(CH2) a-1 CH(X 7 )-, where a is , 2, 3, 4, 5, or 6. X 7 Ga-NR 6 R 7 In some embodiments, R 6 and R 7 are each independently hydrogen or a C1 to C6 alkyl, such as a C1 to C3 alkyl. For example, in some embodiments, X 7 -NH2, -NHCH3, -N HCH2CH3, -NHCH2CH2CH3, -N(CH3)2, -N(CH2CH3) 2, or -N(CH2CH2CH3)2. Alternatively, X 7 Ga-NR 6 R 7 is In some embodiments, R 6 and R 7 together with the nitrogen to which they are attached, or a 4- to 7-membered heterocyclyl optionally substituted with two C1-C6 alkyl groups Alternatively, X 7 Ga-NR 6 R 7 In other embodiments, R 6 and R 7 teeth , together with the nitrogen to which they are attached, form a 4- to 7-membered heterocyclyl. A 7-membered heterocyclyl is a heterocyclic group having one or two C1-C6 alkyl groups, such as a C1-C3 alkyl group. optionally substituted with alkyl groups and / or 4- to 7-membered heterocyclyl optionally containing additional heteroatoms selected from oxygen, sulfur, and nitrogen. For example, in some embodiments, X 6is azetidinyl, methylazetidinyl, Pyrrolidinyl, methylpyrrolidinyl, piperidinyl, methylpiperidinyl, piperazinyl , methylpiperazinyl, dimethylpiperazinyl, morpholinyl, diazepanyl, or methylpiperazinyl It is tildiazepanil.

[0111] In various embodiments, A in formula (I) 1 and A 2 are each independently (1) C5~C 12 Haloalkyl; (2) C5~C 12 Alkenyl; (3) C5~C 12 Alkynyl; (4) (C5~C 12 Alkoxy)-(CH2) n2 -; (5) 1 or 2 halo groups, C1-C6 alkyl groups, C1-C6 haloalkyl groups, or C1-C6 alkoxy groups optionally ring-substituted (C5-C 10 aryl) -(CH2) n3 -; and (6) One or two C1-C6 alkyl groups optionally substituted on the ring (C3-C 8Cycloalkyl)-(CH2) n4 -; Selected from or or alternatively, A 1 and A 2 together with the atoms to which they are bonded, Or two C4-C 10 Forms 5- to 6-membered cyclic acetals substituted with alkyl groups .

[0112] In various embodiments of Formula (I), n1, n2, and n3 are each independently 1 to 4 n4 is an integer between 0 and 4 (i.e., 1, 2, 3, or 4), and n5 is an integer between 0 and 4 (i.e., In various embodiments, A 1 and A 2 is the same It has a chemical structure.

[0113] In various embodiments of formula (I), A 1 and A 2 are C5 to C 12 Ha For example, in various embodiments, C5-C 12 Haloalkyl is a C6 Fluoroalkyl, C7 fluoroalkyl, C8 fluoroalkyl, C9 fluoroalkyl , C 10 Fluoroalkyl, C 11 Fluoroalkyl, or C 12 Fluoroalkyl Which, C5~C 12 Fluoroalkyl. C5~C 12 Halo attached to haloalkyl The number of halogen atoms varies over a wide range depending on the length of the alkyl chain and the degree of halogenation. For example, in various embodiments, C5 to C 12 Haloalkyl is 1-2, such as 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, or 11 halogen atoms In various embodiments, the halogen atom ranges from C5 to C6. 12 Haloalkyl is C F3(CF2) n5 - (where n5 is an integer ranging from 0 to 5), C5-C containing groups 12 For example, in various embodiments, C5 to C 12 Fluoroalkyl is CF3(CF2) n5 (CH2) n6 -, where n5 is , n6 is an integer ranging from 0 to 11, and n5+n6+1 is C5~C12 Equal to the number of carbons in the fluoroalkyl.

[0114] In various embodiments of formula (I), A 1 and A 2 are independently C5 to C 12 The position of the alkenyl double bond may vary. For example, various In the embodiment, C5 to C 12 Alkenyl is CH3CH2CH=CH(CH2)4- etc. CH3CH2CH=CH(CH2) n7 (wherein n7 is an integer ranging from 1 to 8) In some embodiments, C5 to C 12 Alkenyl is, for example, (CH3)2C= CH(CH2) n8 -CH(CH3)-(CH2) n9 - (wherein n8 and n9 are are branched, such as 1, 2, or 3).

[0115] In various embodiments of formula (I), A 1 and A 2 are independently C5 to C 12 The position of the alkynyl triple bond may vary. For example, various In the embodiment, C5 to C 12 Alkynyl is CH3CH2C≡C(CH2)4-, etc. CH3CH2C≡C(CH2) n10 - (wherein n10 is an integer ranging from 1 to 8) In some embodiments, C5 to C 12 Alkynyl is, for example, (CH3)2CHC≡ C(CH2) n11 -CH(CH3)-(CH2) n12 - (wherein n11 and n12 are each independently 1, 2, or 3, and n11 + n12 is in the range of 2 to 5. ) and other branching types.

[0116] In various embodiments of formula (I), A 1 and A 2 are each independently (C5~C1 2alkoxy)-(CH2) n2 In various embodiments, each n2 is independently 1 The position of the oxygen is varied and is an integer in the range of 1 to 4 (i.e., 1, 2, 3, or 4). For example, in various embodiments, (C5 to C 12 Alkoxy)-(CH2) n2 - is CH3O(CH2) such as CH3O(CH2)7- n13 -(CH2) n2 -(formula In another embodiment, n13 is an integer ranging from 1 to 11. 12 Alkoxy)-(CH2) n2 - is CH3(CH2)7-O-(CH2)2-(CH2 ) n2 - CH3 (CH2) n14 -O-(CH2) n15 -(CH2) n2 -( In the formula, n14 and n15 each independently represent an integer between 1 and 8, and n14+n1 and 5 is an integer ranging from 4 to 11. In some embodiments, C5 to C 12 Alcoki For example, CH3O(CH2) n16 -CH(CH3)-(CH2) n17 -(CH 2) n2 - (wherein n16 and n17 each independently represent 1, 2, 3, 4, or 5) and n16+n17 is an integer ranging from 2 to 9), and the like.

[0117] In various embodiments of formula (I), A 1 and A 2are each independently 1 or 2 halo group, C1-C6 alkyl group, C1-C6 haloalkyl group, or C1-C6 alkoxy group optionally ring-substituted with hydroxy groups (C5-C 10 Aryl)-(CH2) n3 -is In various embodiments, each n3 is independently an integer between 1 and 4 (i.e., 1, 2, 3, or is 4). In some embodiments, C5 to C 10 Aryl is phenyl. For example: In various embodiments, (C5 to C 10 Aryl)-(CH2) n3 - 1 or 2 pieces Halo group, C1-C6 alkyl group, C1-C6 haloalkyl group, or C1-C6 alkoxy group C6H5-(CH2) optionally ring-substituted with silyl groups n3 In one embodiment, Optionally ring-substituted (C5-C 10 Aryl)-(CH2) n3 -CF3-C CF3-C6H4-, such as 6H4-CH2- or CF3-C6H4-(CH2)2- (CH2) n3 In another embodiment, an optionally ring-substituted (C5-C 10 Aryl)-(CH2) n3 - is CH3(CH2)3-C6H4-CH2- or CH CH3-(CH2), such as 3(CH2)3-C6H4-(CH2)2- n18 -C6H 4-(CH2) n2 - (wherein n18 is 1, 2, or 3, and n2 is 1, 2, 3) , or 4).

[0118] In various embodiments of formula (I), A 1 and A 2 are each independently 1 or 2 (C3-C8 cycloalkyl)-( CH2) n4 In various embodiments, each n4 is independently an integer between 0 and 4 (including In some embodiments, the C3-C8 cycloalkyl The alkyl is cyclohexyl or cyclopentyl. For example, in various embodiments, (C 3~C8 cycloalkyl)-(CH2) n4 - C6H 11 -(CH2)2-, CH 11 -(CH2)3-, or CH3-C6H 10 1 or 2, such as -(CH2)3- C6H optionally ring-substituted with C1-C6 alkyl groups 11 -(CH2) n4 -in be.

[0119] Alternatively, in other embodiments of formula (I), A 1 and A 2 are the atoms that bind these together. Together with the child, one or two C4-C 10 5-6 membered cyclic ring substituted with alkyl groups For example, in one embodiment, A 1 and A 2 As shown below, this Together with the atoms to which they are attached, they form a six-membered ring substituted with two C8 alkyl groups. Forms acetal.

[0120] [ka] In another embodiment, A 1 and A 2 together with the atoms to which they are bonded, as follows: This forms a five-membered cyclic acetal ring-substituted with two C8 alkyl groups.

[0121] [ka]

[0122] 2. Ionizable lipids In one embodiment, the disclosed lipid nanoparticles comprise an ionizable lipid. Ionizable lipids typically contain an amine-containing group on the head group. In various embodiments, In some embodiments, the ionizable lipid is a compound of formula (I): at 35, 45, 50, or 65 mole percent relative to the total moles of the lipid nanoparticle components In another embodiment, the ionizable lipid is present in the lipid nanoparticle. It is present in about 33 mol % to about 36 mol %, based on the total moles of the components. In this state, the ionizable lipids account for approximately 35 mol% of the total moles of the lipid nanoparticle components. It exists in.

[0123] Further embodiments include: ionizable lipids; phospholipids; polyethylene glycol-lipids cholesterol; and optionally, nucleic acids. In this embodiment, the ionizable lipid is represented by the formula (I), (Ia), (II), (IIa), (IIb), In some embodiments, the ionized The amount of lipid available is approximately 35-65 mole percent based on the total moles of the lipid nanoparticle components. It exists in the range of

[0124] 3. Sterols In some embodiments, the disclosed lipid nanoparticles comprise one or more sterols. In one embodiment, the sterol is cholesterol, or a variant or derivative thereof. In some embodiments, the cholesterol is modified, e.g., oxidized. Unmodified cholesterol is modified by enzymatic action to oxidize the side chain or the ring. Cholesterol can form modified variants on the beta ring structure and may be oxidized on the hydrocarbon tail structure. Exemplary cholesterols that may be used include, but are not limited to, 25-hydroxycholesterol 20α-Hydroxycholesterol (20α-OH), 27- Hydroxycholesterol, 6-keto-5α-hydroxycholesterol, 7-ketocholesterol Sterols, 7β-hydroxycholesterol, 7α-hydroxycholesterol, 7β -25-dihydroxycholesterol, beta-sitosterol, stigmasterol, In one embodiment, the hydroxybenzoates include brassicasterol, campesterol, or a combination thereof. In this study, side-chain oxidized cholesterol was shown to enhance cargo delivery compared to other cholesterol variants. In one embodiment, the cholesterol is unmodified cholesterol. .

[0125] 4. PEG-lipids In some embodiments, the disclosed nanoparticle compositions also contain one or more PEG or includes PEG-modified lipids. Such lipids may alternatively be referred to as PEGylated lipids or PEG-lipids. The inclusion of PEGylated lipids can be used to develop lipid nanoparticles in vitro. It can enhance the stability and circulation time of the polymer colloid in vivo. In embodiments, PEGylation is advantageous in that the PEG moiety is slowly released in the blood circulation. Exemplary PEG-lipids include, but are not limited to, C6-C20 of Included are PEG conjugated to saturated or unsaturated alkyl chains of any length. PEG-modified phosphatidylethanolamine, PEG-modified phosphatidic acid, PEG -Modified ceramide (PEG-CER), PEG-modified dialkylamine, PEG-modified dialkylamine dialkylglycerol (PEG-DAG), PEG-modified dialkylglycerol, and For example, the PEG lipid may be a mixture of PEG-c-DOMG, PEG-DMG, PEG -DLPE, PEG-DMPE, PEG-DPPE, PEG-DSG, or PEG-D Each lipid in the SPE can be used.

[0126] 5. Phospholipids The phospholipid component of the nanoparticles may be one or more (poly)unsaturated lipids. The phospholipids can be assembled into one or more lipid bilayers. In some embodiments, the phospholipid may comprise a phospholipid moiety and one or more The fatty acid moiety may include:

[0127] In some embodiments, the phospholipid moiety includes, but is not limited to, phosphatidylcholine. Phosphorus, Phosphatidylethanolamine, Phosphatidylglycerol, Phosphatidyl Serine, phosphatidic acid, 2-lysophosphatidylcholine, and sphingomyelin In some embodiments, the fatty acid moiety includes, but is not limited to, lauric acid. Acid, myristic acid, myristoleic acid, palmitic acid, palmitoleic acid, stearic acid , oleic acid, linoleic acid, alpha-linolenic acid, erucic acid, phytanic acid, arachidonic acid Acid, arachidonic acid, eicosapentaenoic acid, behenic acid, docosapentaenoic acid, and docosapentaenoic acid Modifications and modifications, including branching, oxidation, cyclization, and alkynes, are available. Non-naturally occurring species, including naturally occurring species, with substitutions are also contemplated. For example, phospholipids may contain one or more is a compound with multiple alkynes (e.g., one or more double bonds replaced by triple bonds). The alkenyl groups may be functionalized with or crosslinked to the alkyl groups. Under the right reaction conditions, the alkyne group undergoes copper-catalyzed cycloaddition when exposed to azides. Such reactions may result in the formation of nanoparticle compositions to facilitate membrane penetration or cell recognition. on the lipid bilayer functionalization, or on targeting or imaging moieties (e.g. The nanoparticle compositions may be useful for conjugating the nanoparticle compositions to useful moieties such as dyes. It is possible.

[0128] Exemplary phospholipids include, but are not limited to, 1,2-distearate 1,2-Dioleoyl-sn-glycero-3-phosphocholine (DSPC), 1,2-Dioleoyl-sn -Glycero-3-phosphoethanolamine (DOPE), 1,2-dilinoleoyl-sn -glycero-3-phosphocholine (DLPC), 1,2-dimyristoyl-sn-glycero -phosphocholine (DMPC), 1,2-dioleoyl-sn-glycero-3-phosphocholine Dipalmitoyl-sn-glycero-3-phosphocholine (DOPC), 1,2-dipalmitoyl-sn-glycero-3-phosphocholine (DPP C), 1,2-diundecanoyl-sn-glycero-phosphocholine (DUPC), 1-pa Lumitoyl-2-oleoyl-sn-glycero-3-phosphocholine (POPC), 1,2 -Di-O-octadecenyl-sn-glycero-3-phosphocholine (18:0 diether P C), 1-oleoyl-2-cholesterylhemisuccinoyl l)-sn-glycero-3-phosphocholine (OChemsPC), 1-hexadecyl-s n-Glycero-3-phosphocholine (C16 lyso-PC), 1,2-dilinolenoyl-sn -glycero-3-phosphocholine, 1,2-diarachidonoyl-sn-glycero-3-phosphocholine 1,2-didocosahexaenoyl-sn-glycero-3-phosphocholine, 1, 2-Diphytanoyl-sn-glycero-3-phosphoethanolamine (ME 16.0 PE), 1,2-distearoyl-sn-glycero-3-phosphoethanolamine, 1, 2-Dilinoleoyl-sn-glycero-3-phosphoethanolamine, 1,2-Dilinoleoyl Noyl-sn-glycero-3-phosphoethanolamine, 1,2-diarachidonoyl-s n-Glycero-3-phosphoethanolamine, 1,2-didocosahexaenoyl-sn- Glycero-3-phosphoethanolamine, 1,2-dioleoyl-sn-glycero-3- Phospho-rac-(1-glycerol) sodium salt (DOPG), dipalmitoyl phosphatase Photidylglycerol (DPPG), palmitoyl oleoyl phosphatidylethanolamine POPE, distearoyl-phosphatidyl-ethanolamine (DSPE) ), dipalmitoylphosphatidylethanolamine (DPPE), dimyristoylphosphatidylethanolamine (DPPE) Diethanolamine (DMPE), 1-stearoyl-2-oleoyl-phosphatidyl Ethanolamine (SOPE), 1-Stearoyl-2-oleoyl-phosphatidylcholine Phosphingomyelin (SOPC), sphingomyelin, phosphatidylcholine, phosphatidylethanolamine cholamine, phosphatidylserine, phosphatidylinositol, phosphatidic acid, Palmitoyloleoylphosphatidylcholine, Lysophosphatidylcholine, Lysophosphatidylcholine In a preferred embodiment, the phospholipid is DSPC. In another embodiment, the phospholipid is DMPC.

[0129] E. Cargo In one embodiment, the disclosed lipid nanoparticle compositions can be used to deliver a therapeutic agent or In some embodiments, the therapeutic or prophylactic agent is delivered by the lipid nanoparticle. In one embodiment, the lipid nanoparticles are loaded with one or more nucleic acids. are.

[0130] Representative nucleic acids include, but are not limited to, deoxyribonucleic acid (DNA), ribonucleic acid (RI), (RNA)RNA, DNA, single-stranded RNA, single-stranded DNA, double-stranded RNA, double-stranded DNA , triple-stranded DNA, siRNA, shRNA, sgRNA, mRNA, miRNA, and In one embodiment, the nucleic acid is an siRNA, miRNA, mRNA, or RNA, expressed DNA, antisense oligonucleotides, or immunostimulatory oligonucleotides It's a punchline.

[0131] CRISPR (clustered regularly interspaced short palindromic repeats)-based Gene editing requires two components: guide RNA and CRISPR-associated endonuclease. The guide RNA directs the Cas nuclease to a specific target. Cas directs itself to a DNA sequence and then creates a double-strand break in the DNA at that site. In one embodiment, the disclosed lipid nanoparticles can be used to perform CRISPR-based gene editing. In one lipid nanoparticle, the nucleic acid cargo is a guide RNA. A. In such an embodiment, the second lipid nanoparticle comprises an RNA-guided endonuclease. The two types of lipid nanoparticles can be administered together and contain a nucleic acid cargo encoding the enzyme. Exemplary RNA-guided endonucleases include, but are not limited to, However, it includes Cas9, CasX, CasY, Cas13, or Cpf1.

[0132] In one embodiment, the cargo is an siRNA. Induce sequence-specific post-transcriptional gene silencing, thereby reducing or In one example, siRNA is a double-stranded RNA that can inhibit or even block a gene. Within the region of sequence identity between both the target RNA and the homologous RNA molecule, such as an mRNA, For example, WO 02 / 44321 discloses a method for producing a 3' overhang. We have developed siRNAs that can induce sequence-specific degradation of target mRNAs when base-paired with their termini. and methods for making these siRNAs are incorporated herein by reference. Sequence-specific gene silencing is achieved by siRNAs generated by the enzyme Dicer. This can be done in mammalian cells using synthetic, short double-stranded RNA that mimics (Elbashir, et al. (2001) Nature, 411:494 498)(Ui-Tei, et al. (2000) FEBS Lett 479:79-82).

[0133] In one embodiment, the cargo is messenger RNA (mRNA). -RNA uses the cell's protein synthesis machinery to synthesize it after the cargo reaches the cytoplasm. It is a single-stranded RNA that can be translated into the protein it encodes. Qualitatively, the 5'UTR serves to recruit RNA-binding proteins and microRNAs and may be affected by modifications in the 3'UTR, and RNA-binding proteins and All microRNAs can affect translation activity (Sahin, et al. (2014) Na t Rev Drug Disco 13: 759-580) (Kariko et al (2008) Mol Ther 11:1833-1840).

[0134] In one embodiment, the lipid nanoparticles contain less than 1.0 mg / kg of inhibitory nucleic acid. The particles were prepared at 1.0, 0.9, 0.8, 0.7, 0.6, or 0.5 mg / kg of inhibitory nucleic acid. In another embodiment, the lipid nanoparticles may contain 0.5 mg / kg, which is an advantage over current technology, where nanoparticles requires high doses of nucleic acid (>1 mg / kg) to achieve gene silencing, The doses are not approved for human delivery. The disclosed technology involves targeting ligands. Gene silencing was achieved using 0.5 mg / kg of inhibitory nucleic acid in lipid nanoparticles containing no ATP. This can be achieved.

[0135] In some embodiments, nucleic acids, including but not limited to oligonucleotides, are stable. modified or modified to improve quality, half-life, and / or nuclease stability or another modified nucleotide. Phosphodiester oligodeoxyribonucleotides, natural phosphodiester oligoribonucleotides Nucleotides, ribonucleotide polymers, and deoxyribonucleotide polymers are , and another different modification. Exemplary modifications include, but are not limited to: However, phosphorothioate (PS) bonds, 2'-O-methyl (2'OM) e), 2' fluoro bases, inverted dT and ddT, phosphorus at the 3' end of the oligonucleotide Oxidized, locked nucleic acids are included, and phosphoramidite C3 spacers are included.

[0136] Phosphorothioate bonds are bonds in the phosphate backbone of oligonucleotides. In place of the non-bridging oxygen, a sulfur atom is used. Approximately 50% of The PS modification results in the formation of two stereoisomers, which allows the internucleotide linkage to be In some embodiments, the nucleic acid is One or more PS bonds, e.g., 5' and 6', to inhibit xonuclease degradation. The oligonucleotides contain at least three PS bonds at the 3' and 4' ends. It also helps reduce attack by endonucleases by cleaving the entire oligonucleotide. and contains PS bonds.

[0137] 2'OMe, a naturally occurring post-transcriptional modification of RNA, is a cleavage site of tRNA and other small RNAs. In some embodiments, the nucleic acid or oligonucleotide contains 2'OMe. This modification improves the Tm of the RNA:RNA duplex, but Only minor changes in RNA:DNA stability are made. This modification is due to the single-stranded endonuclease In some embodiments, the nucleotide sequence protects against attack by nucleotides containing nucleotides but not against exonuclease digestion. Such nucleic acids or oligonucleotides may also be end-blocked. DNA oligonucleotides containing 5′-diaminobenzyl methyltransferase are generally less susceptible to DNase activity than unmodified DNA. 2'OMe modifications improve stability and enhance binding affinity to target transcripts. It is often used in antisense oligonucleotides as a means of improving compatibility. There are.

[0138] The 2'-fluoro base has a fluorine-modified ribose, which increases the binding affinity (Tm). It also confers some relative nuclease resistance compared to native RNA. In some embodiments, the nucleic acid or oligonucleotide is provided in combination with a PS-modified linkage. Contains a 2' fluoro base.

[0139] An inverted dT can be incorporated at the 3' end of an oligonucleotide, resulting in 3' exonucleolytic 3'-3', which inhibits degradation by ATPase and extension by DNA polymerase In addition, an inverted 2',3' didethiol is attached to the 5' end of the oligonucleotide. Spurious ligation is prevented by placing an oxy-dT base (5' inverted ddT) and can protect against some forms of enzymatic degradation.

[0140] Some embodiments include nucleic acids or oligonucleotides containing a phosphoramidite C3 spacer. The phosphoramidite C3 spacer can also be incorporated internally, Alternatively, fluorophores or other pendant groups can be incorporated at either end of the oligo. A long hydrophilic spacer arm can also be introduced for attachment of the C3 spacer. It is also possible to suppress degradation by 3' exonucleases by using a 3' exonuclease.

[0141] In some embodiments, the nucleic acid or oligonucleotide comprises a locked nucleic acid. The 2'-O and 4'-C atoms of ribose are linked to the acid through a methylene bridge. This additional bridge is usually associated with a flexible RNA nucleotide. LNAs are a class of molecules that constrain the structure of RNA and DNA, essentially locking it into a rigid conformation. The oligonucleotide may be inserted into both the nucleotides.

[0142] Other types of cargo that can be delivered via the disclosed nanoparticles include those containing These include, but are not limited to, chemotherapeutic agents, cytotoxic drugs, radioactive ions, small molecules, proteins, poly It includes oligonucleotides and nucleic acids.

[0143] Representative chemotherapeutic agents include, but are not limited to: amsacrine; Bleomycin, busulfan, capecitabine, carboplatin, carmustine, chloramphenicol Mubucil, cisplatin, cladribine, clofarabine, crisantaspase, cyclo Phosphamide, cytarabine, dacarbazine, dactinomycin, daunorubicin, docetamol doxorubicin, epirubicin, etoposide, fludarabine, fluorouracil , gemcitabine, hydroxycarbamide, idarubicin, ifosfamide, irinotecan , leucovorin, liposomal doxorubicin, liposomal daunorubicin, lomustine, Melphalan, mercaptopurine, mesna, methotrexate, mitomycin, mitoxantrone Santrone, oxaliplatin, paclitaxel, pemetrexed, pentostatin, Rocarbazine, raltitrexed, satraplatin, streptozocin, tegafur Rasil, temozolomide, teniposide, thiotepa, thioguanine, topotecan, treosul Fen, vinblastine, vincristine, vindesine, vinorelbine, or any of these Exemplary pro-apoptotic agents include, but are not limited to, fludarabine tau fludarabinetaurosporine, cycloheximide, actinomycin D, These include ctosylceramide, 15d-PGJ(2), and combinations thereof.

[0144] Some embodiments provide a method of delivering a nucleic acid to a subject in need thereof, comprising: and a method comprising administering the lipid nanoparticle composition to a subject. In this context, nucleic acids include siRNA, miRNA, mRNA, expressed DNA, and antisense oligos. nucleotides, or immunostimulatory oligonucleotides.

[0145] B. Exemplary Lipid Nanoparticle Formulations In one embodiment, the lipid nanoparticle formulation comprises about 30 mol % to about 70 mol % of an ionizable lipid. ol%, phospholipids approx. 5 mol% to approx. 25 mol%, cholesterol approx. 25 mol% to approx. 4 5 mol%, and about 0 mol% to about 5 mol% PEG-lipid. The lipid nanoparticle formulation contains approximately 45 mol% ionizable lipids, approximately 9 mol% phospholipids, and 10 mol% hydroxybenzoates. In another embodiment, the composition comprises about 44 mol% cholesterol and about 2 mol% PEG-lipid. The lipid nanoparticle formulation contains approximately 50 mol% ionizable lipids, approximately 9 mol% phospholipids, It contains about 38 mol% cholesterol and about 3 mol% PEG-lipid.

[0146] One embodiment comprises about 40 moles of an ionizable lipid of formula (I) relative to the total moles of the following four components: mol% to approximately 60mol%, 1-2-distearoyl-sn-glycero-3-phosphocholine Approximately 5 mol% to approximately 15 mol% of C 14 PEG 2000 Approximately 1 mol% to approximately 5 mol%, and about 30 mol% to about 47 mol% cholesterol. do.

[0147] Another embodiment is a mixture of 50 moles of ionizable lipid of formula (I) relative to the total moles of the following four components: mol%, 1-2-distearoyl-sn-glycero-3-phosphocholine 9mol%, C 14 PEG 2000 Lipid nanoparticles containing 3 mol% of cholesterol and 38 mol% of cholesterol Substitute formulations are provided.

[0148] Another embodiment is a lipid-binding protein comprising a lipid-binding protein (ionizable lipid, cholesterol, lipid-PEG, and phosphorus). The lipid nanoparticle formulation has a lipid:mRNA mass ratio of between about 2:1 and 50:1. .

[0149] C. Pharmaceutical Compositions Pharmaceutical compositions comprising the disclosed lipid nanoparticles are provided. The lipid nanoparticle compositions are generally The pharmaceutical composition may be formulated in whole or in part as a pharmaceutical composition. For example, but not limited to, a pharmaceutical composition may include multiple nanoparticle compositions. However, one or more different therapeutic agents containing one or more nucleic acids of different types and and / or a prophylactic agent, and In some embodiments, the pharmaceutical composition may include One or more pharmaceutically acceptable ingredients, including but not limited to a pharmaceutically acceptable carrier. Contains excipients or accessory ingredients.

[0150] Pharmaceutical compositions containing nanoparticles can be administered parenterally (intramuscularly, intraperitoneally, intravenously (IV) or dermally). injection below), transdermal (either passively or using iontophoresis or electroporation) or by transmucosal (nasal, vaginal, rectal or sublingual) administration route, or bioerodible It can be formulated for administration by using an insert and can be used for each route of administration. It can be formulated into an appropriate dosage form.

[0151] In some in vivo approaches, the nanoparticle compositions disclosed herein are used to treat As used herein, "effective amount" or "therapeutically effective amount" refers to a therapeutically effective amount of a compound administered to a subject. The term "treatment" refers to the treatment of a disorder that is caused by treating, suppressing, or alleviating one or more symptoms of the disorder being treated. or a dosage sufficient to provide the desired pharmacological and / or physiological effect. The exact dosage will depend on subject-dependent variables (e.g., age, health of the immune system, etc.). etc.), the disease, and the treatment being performed.

[0152] As further studies are conducted on the disclosed nanoparticles, various therapeutic effects in various patients will be evaluated. Information is emerging and those skilled in the art will be able to determine appropriate dosage levels for treating the condition. , to ensure appropriate medication, taking into account the recipient's medical status, age, and overall health. The selected dosage will depend on the desired therapeutic effect, the route of administration, and the desired For the disclosed nanoparticles, the dose is generally 1 kg of body weight per day. Dosage levels of 0.001 mg to 5 mg of nucleic acid per gram are administered to mammals. In the present study, the preferred dose of the disclosed nanoparticles is 0.01 mg / kg to 0.25 mg / kg. The disclosed nanoparticles generally contain four components: ionizable lipids, cholesterol, and Dosage range: 0.2 mg to 100 mg / kg body weight (including PEG-lipids, PEG-lipids, and phospholipids) A dose of the disclosed nanoparticles is administered to a mammal. More specifically, the preferred dose of the disclosed nanoparticles is 0. .05mg / kg~0.5mg / 4 ingredients 1kg / 1kg body weight.

[0153] In certain embodiments, the lipid nanoparticle composition is injected directly into the site to be treated, e.g., Typically, it can be administered systemically by injection. This results in an increased local concentration of the lipid nanoparticle composition that is greater than would be possible without the lipid nanoparticle composition. The porous nanoparticle composition is combined with the matrix to form a porous nanoparticle that is transported away from the site to be treated. Reducing passive diffusion of the polypeptide creates an increased local concentration of the polypeptide composition. It can help to get it out.

[0154] 1. Formulations for parenteral administration In some embodiments, the nanoparticle compositions disclosed herein contain lipid nanoparticles. The formulations are also administered in aqueous solutions, including suspensions or emulsions, by parenteral injection. Generally, the pharmaceutical composition contains an effective amount of lipid nanoparticles. and optionally containing pharmaceutically acceptable diluents, preservatives, solubilizers, emulsifiers, etc. Such compositions optionally contain: The composition may contain one or more of the following: diluent, sterile water, various buffer contents (e.g., Tris-HCl l, acetate, phosphate), pH and ionic strength buffered saline; and surfactants and solubilizers (e.g., TWEEN 20 (polysorbate-20), TWEEN 8 0 (Polysorbate-80)), antioxidants (e.g., ascorbic acid, sodium metabisulfite sodium), and preservatives (e.g., thimerosal, benzyl alcohol), and bulking agents Additives such as sugars (e.g., lactose, mannitol). Examples of non-aqueous solvents or vehicles are propylene glycol, polyethylene glycol, olive oil and corn oil, etc. Vegetable oils, gelatin, and injectable organic esters such as ethyl oleate. It can be lyophilized and redissolved / resuspended immediately before use. The composition can be irradiated by incorporating a sterilizing agent into the composition by filtration through a filter. The composition can be sterilized by irradiation or by heating the composition.

[0155] 2. Controlled Delivery Polymer Matrix The lipid nanoparticles disclosed herein can also be administered in controlled release formulations. The polymer device is a polymer device (rod, cylinder, film, disc) They can be engineered for systemic long-term release after implantation or injection (microparticles). The matrix can be in the form of microparticles, such as microspheres. However, in this case, the drug is dispersed within a solid polymer matrix or microcapsules. , where the core is made of a different material than the polymer shell and the peptide is essentially liquid. The particles are dispersed or suspended in a core which may be solid or inorganic. Unless otherwise specified, microparticles, microspheres, and microcapsules are used interchangeably. Alternatively, the polymer can be formed into thin slabs or layers ranging in size from a few nanometers to 4 centimeters. may be a film, a powder produced by grinding or other standard techniques, or even The composition can be cast as a gel, such as a hydrogel.

[0156] Use of either non-biodegradable or biodegradable matrices for delivery of lipid nanoparticles Although biodegradable matrices are possible, in some embodiments biodegradable matrices are preferred. The polymers may be either polymers or synthetic polymers, but are characterized by their degradation and release profiles. Synthetic polymers are preferred in some embodiments due to their better performance ratings. The polymer is selected based on the period of time over which release is desired. Linear release may be most useful. In some cases, pulsed or "bulk" release may provide more effective results. The polymer can be in the form of a hydrogel (typically up to about 90% by weight). in absorbing water), and can be crosslinked with multivalent ions or polymers, if desired. .

[0157] The matrix can be prepared by solvent evaporation, spray drying, solvent extraction and other methods known to those skilled in the art. Bioerodible microspheres can be formed by methods such as those described in Mathiowitz et al. nd Langer, J. Controlled Release, 5:13-22 (1987);Mathiowitz, et al., Reactive P olymers, 6:275-283 (1987); and Mathiowitz, et al., J. Appl. Polymer Sci., 35:7 55-774 (1988) developed to produce microspheres for drug delivery. The compound can be prepared using any of the methods described above.

[0158] The device is for local release to treat the area of ​​implantation or injection - this is usually not systemic This results in the delivery of a much smaller dose than that used for general treatment - or systemic delivery. These can be implanted subcutaneously into muscle, fat, or It can be injected or swallowed.

[0159] D. Methods for Producing Lipid Nanoparticles Methods for producing lipid nanoparticles are known in the art. The lipid nanoparticles are fabricated using microfluidics. For an exemplary method of forming lipid nanoparticles using lipid nanoparticles, see Leung, AKK, et al. l., J Phys Chem, 116:18440-18450 (2012), Chen, D., et al., J Am Chem Soc, 134:69 47-6951 (2012), and Belliveau, NM, et al., Molecular Therapy- Nucleic Acids, 1: e37 (2012). In summary, The lipid nanoparticles are prepared in a buffer. Other lipid nanoparticle components (ionizable lipids, PEG -lipids, cholesterol, and DSPC) are prepared in separate buffers. The two solutions are introduced into the microfluidic device by a pump. They contact within a fluidic device to form lipid nanoparticles that encapsulate the cargo.

[0160] The disclosed method for screening lipid nanoparticles is described in International Patent Application PCT / US / 2002 / 002444. No. 018 / 058171, which is incorporated by reference in its entirety. The screening method identifies vehicle delivery formulations Characterize and deliver functional cargo with desired tropism to the cytoplasm of specific cells. The screening method identifies formulations that detect when delivered to cells. When reporter function is detected in cells, This means that the delivery vehicle can be formulated to deliver functional cargo to cells. The chemical composition identifier is used to indicate that the formulation of the different delivery vehicles This includes constantly tracking the unique chemical composition of each different delivery vehicle formulation. In an embodiment, the chemical composition identifier is a nucleic acid barcode. Pairing the chemical components used to formulate the delivery vehicle loaded with When the nucleic acid barcodes were sequenced, the chemical composition of the delivery vehicle that delivered the barcodes Representative reporters include, but are not limited to, siRNA, mRNA, A, nuclease protein, nuclease mRNA, small molecule, epigenetic modification factors, and phenotype-modifying factors.

[0161] E. Method of Use Lipid nanoparticles disclosed for delivering cargo, e.g., nucleic acids, to specific cells or organs Disclosed herein are methods of using the nanoparticles. , in the absence of a targeting ligand, specific cells or In another embodiment, the disclosed lipid nanoparticles are , are useful for treating or preventing disease in a subject in need thereof.

[0162] In some embodiments, the disclosed nanoparticles are delivered directly to a subject. The lipid nanoparticles are contacted with cells ex vivo, and the treated cells are administered to a subject. The cells are differentiated into autologous cells, such as, but not limited to, T cells or T cells. In some embodiments, the disclosed lipid nanoparticles can be used to treat or treat various types of cancer, including cancer cells, which can be immune cells. The particles can be used as vehicles for adoptive cell transfer.

[0163] 1. Methods for delivering cargo into cells Methods for delivering therapeutic and / or prophylactic nucleic acids to a subject in need thereof are described herein. provided in the book.

[0164] In some embodiments, the disclosed lipid nanoparticle compositions comprise a particular type or class of lipid nanoparticles. Target cells (e.g., cells of a particular organ or system). For example, and / or prophylactic agents, specifically to immune cells of a subject. Exemplary immune cells include, but are not limited to, CD8+, CD4+, or In another embodiment, the lipid nanoparticles are administered to target In the absence of a binding ligand, mammalian liver hepatocytes, liver immune cells, splenic T cells, or lung It can be formulated for delivery to endothelial cells. Specificity of delivery to target areas indicates that a higher proportion of lipid nanoparticles are specific to the target type or class. In some embodiments, the specificity of the delivery allows for Therefore, the amount of therapeutic and / or prophylactic agent delivered per gram of target tissue It can result in a 2-fold, 5-fold, 10-fold, 15-fold, or even 20-fold increase.

[0165] 2. Methods of gene regulation Provided herein are methods of using the disclosed lipid nanoparticles for gene regulation. In one embodiment, the lipid nanoparticles are capable of inducing gene expression in target cells in a subject in need thereof. Lipid nanoparticles can be used to reduce the incidence of targeting ligands. The inhibitory nucleic acid can be delivered to a target cell of interest without the need for a siRNA. It can be said that:

[0166] Another embodiment provides the disclosed method for editing a gene in a cell of a subject in need thereof. and methods of using the lipid nanoparticles.

[0167] In one embodiment, the cells targeted for gene regulation are immune cells. These may be T cells, such as CD8+ T cells, CD4+ T cells, or regulatory T cells. Other exemplary immune cells for gene editing include, but are not limited to, macrophages, macrophages, and oocytes. In some embodiments, the IL-16 receptor agonist (IL-16) may be a phage, a dendritic cell, a B cell, or a natural killer cell. In this case, the cells targeted for gene regulation are hepatocytes.

[0168] Exemplary genes that can be targeted include, but are not limited to, the following: T cell receptor, B cell receptor, CTLA4, PD1, FOXO1, FOXO3, AK T, CCR5, CXCR4, LAG3, TIM3, killer immunoglobulin-like receptor, GI TR, BTLA, LFA-4, T4, LFA-1, Bp35, CD27L receptor, TNF RSF8, TNFRSF5, CD47, CD52, ICAM-1, LFA-3, L-Sele Cutin, Ki-24, MB1, B7, B70, M-CSFR, TNFR-II, IL-7 R, OX-40, CD137, CD137L, CD30L, CD40L, FasL, TR AIL, CD257, LIGHT, TRAIL-R1, TRAILR2, TRAIL-R 4, TWEAK-R, TNFR, BCMA, B7DC, BTLA, B7-H1, B7-H 2, B7-H3, ICOS, VEGFR2, NKG2D, JAG1, GITR, CD4, CCR2, GATA-3, MTORC1, MTORC2, RAPTOR, GATOR, F OXP3, NFAT, IL2R, and IL7. Other exemplary genes that can be targeted Genes include, but are not limited to, OCT, G6Pase, Mut, PCCA, PCC B, and PAHs.

[0169] Exemplary tumor-associated antigens that T cells can recognize and are intended to target Antigens include, but are not limited to, MAGE1, MAGE3, MAGE6, BAGE, GAGE, NYESO-1, MART1 / Melan A, MC1R, GP100, Ciro Synase, TRP-1, TRP-2, PSA, CEA, Cyp-B, Her2 / Neu, hTERT, MUC1, PRAME, WT1, RAS, CDK-4, MUM-1, KRA S, MSLN and β-catenin.

[0170] 3. Subject to treatment In some embodiments, the subject being treated is a patient with cancer, an autoimmune disease, an infectious disease, an organ transplant, or an organ transplant. The mammal is experiencing organ failure, protein deficiency, or a combination thereof. In some embodiments, the subject is a human. In some embodiments, the methods described herein involve the use of hepatocytes. In some embodiments, the proteins described herein can be translated by the The described methods may be used to administer one or more DNA, mRNA, sgRNA, or siRNA to a liver It can be used to deliver to cells. [Example]

[0171] General Notes: All reactions were carried out using anhydrous solvents unless otherwise stated. The reaction proceeded under nitrogen atmosphere in a flask or vial with magnetic stirring. was purchased from Sigma-Aldrich and used as received. The chromatograph was performed using pre-packed Biotage Sfar silica gel cartridges. Equipped with Biotage Selekt or Teledyne-Isco Combif The analysis was performed using a lash Nextgen300+. Merck Silica Gel 60 Precipitator Thin layer chromatography was performed using iodine and the compounds were visualized using iodine. Nuclear magnetic resonance (NMR) spectroscopy was performed using a Varian INOVA 500 MHz spectrometer. Chemical shifts are the residual solvent peak of CHCl3 at δ = 7.26 ppm. expressed in δ parts per million (ppm) relative to the direction of the high magnetic field of tetramethylsilane Liquid chromatography-mass spectrometry (LCMS) was performed using a Waters Acquisition UPLC BEH C18 column (130Å, 1.7 μM, 2.1 mm × 50 mm) Waters Acquity UPLC with QDa detector (ESI) equipped with The analysis was carried out using an H-class Plus. One of the following basic LCMS methods was used: Compounds were analyzed using Method A: Solvent A = water + 0.1% formic acid, Solvent B = acetonitrile ;Gradually from 90% A, 10% B to 5% A, 95% B in 3 minutes, then hold at 95% B for 2 minutes, and then ramp back to 10% B in 1 minute; flow rate = 0.5 mL / min. Method B: Column - XTERRA RP18 (4.6×50 mm), 5 μ, (Mobile phase: initially 50 % [water with 0.1% HCOOH] and 50% [(70:30) ACN:THF with 0.1% HCOOH]; then at 2.65 minutes, 2% [water with 0.1% HCOOH] and 98% [(70:30) ACN:THF with 0.1% HCOOH], hold this mobile phase composition until 3.75 minutes, and finally at 4.90 minutes, return to the initial conditions, i.e., 50% [water with 0.1% HCOOH] and 50% [(70:30) ACN:THF with 0.1% HCOOH], hold this mobile phase composition until 5.10 minutes. Flow rate = 1.2 ml / min.

[0172] List of Abbreviations[[ID=二十一]] [[ID=二十二]]DCM: Dichloromethane[[ID=二十三]] [[ID=二十四]]DIPEA: N,N - Diisopropylethylamine[[ID=二十五]] [[ID=二十六]]DMAP: 4 - (Dimethylamino)pyridine[[ID=二十七]] [[ID=二十八]]DMPC: 1,2 - Dimyristoyl - sn - glycero - 3 - phosphocholine[[ID=二十九]] [[ID=三十]]DSPC: 1,2 - Distearoyl - sn - glycero - 3 - phosphocholine[[ID=三十一]] [[ID=三十二]]EDC: N - (3 - Dimethylaminopropyl)-N’ - ethylcarbodiimide hydrochloride[[ID=三十三]] [[ID=三十四]]Eq: Equivalent[[ID=三十五]] [[ID=三十六]]ESI: Electrospray Ionization [[ID=三十七]] [[ID=三十八]]LCMS: Liquid Chromatography - Mass Spectrometry[[ID=三十九]]<00022​​​​​​​​​​​​

[0174] Representative synthetic procedures include: Example 1: 3-((4,4-bis(((Z)-oct-5-ene) -1-yl)oxy)butanoyl)oxy)-2-((((3-(diethylamino)propyl (9Z,12Z)-octadeca-9,1 The preparation of the 2-dienoate is exemplified.

[0175] Step 1: 4,4-bis(((Z)-oct-5-en-1-yl)oxy)butane Nitrile (Intermediate Ia)

[0176] [ka]

[0177] The following is representative of general procedure A: Pyridinium p-toluenesulfonate (0 4,4-diethoxybutanediol (0.12g, 0.48mmol, 0.05Eq) was added to a vial containing Tanninnitrile (1.5 g, 9.5 mmol, 1 equiv.) and cis-5-octene-1- The vial was tightly capped and The resulting mixture was heated at 105° C. for 72 hours. After this time, the mixture was cooled to room temperature. The crude material was purified by flash column chromatography (100 g silica, 25 min. hexanes). The product was purified by 0-100% dichloromethane in ethanol. (5-en-1-yl)oxy)butanenitrile (1.14 g, 37%) as a colorless oil was obtained as. 1 H NMR (500 MHz, chloroform-d) δ 5.43 - 5.27 (m, 4H), 4.56 (t, J = 5.3 Hz, 1H), 3.61 (dt, J = 9.3, 6.6 Hz, 2H), 3.44 (dt, J = 9.3, 6.6 Hz, 2H), 2 .42 (t, J = 7.4 Hz, 2H), 2.12 - 1.91 (m, 9H), 1.66 - 1.54 (m, 5H), 1.49 - 1.36 ( m, 4H), 0.96 (t, J = 7.6 Hz, 6H).

[0178] Step 2: 4,4-bis(((Z)-oct-5-en-1-yl)oxy)butane Acid (Intermediate IIa)

[0179] [ka]

[0180] The following is representative of General Procedure B: 4,4-bis(((Z)-oct-5-ene) (1-1-yl)oxy)butanenitrile (Intermediate Ia, 1.14 g, 3.54 mmol, 1 In a vial containing potassium hydroxide (0.60 g, 10.6 mmol, 3 Eq), Ethanol (3.5 mL) and water (3.5 mL) were then added. The mixture was then capped and heated to 110°C for 18 hours. After this time, the mixture was allowed to cool to room temperature. The mixture was diluted with ethyl acetate (20 mL) and 1 M HCl was added to The pH was adjusted to about 5. The resulting biphasic mixture was separated and the aqueous phase was extracted with ethyl acetate (2×2 The organic extracts were combined, dried over sodium sulfate, and filtered. and concentrated to give 4,4-bis(((Z)-oct-5-en-1-yl)oxy)butane The acid (1.16 g, 96% yield) was obtained as a sticky white solid. 1 H NMR (400 MHz, chloroform-d) δ 5.41 - 5.24 (m, 4H), 4.45 (t, J = 5.6 Hz, 1H), 3.51 (dt, J = 9.0, 6.7 Hz, 2H), 3.39 (dt, J = 9.0, 6.7 Hz, 2H), 2.17 (t, J = 7.6 Hz, 2H), 2.08 - 1 .98 (m, 8H), 1.81 (q, J = 7.3 Hz, 2H), 1.59 - 1.52 (m, 4H), 1.44 - 1.32 (m, 4H), 0.94 (t, J = 7.5 Hz, 6H).

[0181] Step 3: 3-Hydroxy-2-(hydroxymethyl)propyl(9Z,12Z)- Octadeca-9,12-dienoate (Intermediate III)

[0182] [ka]

[0183] Trimethylolmethane (3.0 g, 1 Eq, 28 mmol) was dissolved in dichloromethane (100 mL), linoleic acid (7.9 g, 1 Eq, 28 mmol), DIPE A (5.5 g, 7.4 mL, 1.5 Eq, 42 mmol), and DMAP (0.69 g Finally, EDC (8.1 g, 1.5 Eq, 4 2 mmol) was added and stirred at 23° C. for 18 hours. After this time, the reaction mixture was concentrated , flash column chromatography (silica 200 g, hexane over 20 min) Purified by 0-90% ethyl acetate. Propyl (9Z,12Z)-octadeca-9,12-dienoate (2.6g, 25%) was obtained as a colorless oil. 1H NMR (500 MHz, chloroform-d) δ 5.43 - 5.27 (m, 5H), 4.24 (d, J = 6.3 Hz, 2H), 3.76 (ddt, J = 21.1, 11.1, 5.3 Hz, 4H), 2.77 (d, J = 6.8 Hz, 2H), 2.61 - 2.55 (m, 2H), 2.36 - 2.29 (m, 2H), 2.10 - 1.98 (m, 6H), 1.66 - 1.58 (m, 2H), 1.41 - 1.21 (m, 12H), 0.92 - 0.85 (m, 3H).

[0184] Step 4: 3-((4,4-bis(((Z)-oct-5-en-1-yl)oxy) )butanoyl)oxy)-2-(hydroxymethyl)propyl(9Z,12Z)-octa Deca-9,12-dienoate (Intermediate IVa).

[0185] [ka]

[0186] The following is representative of general procedure C: 4,4-bis(4,4-biphenyl) in dichloromethane (10 mL) bis(((Z)-oct-5-en-1-yl)oxy)butanoic acid (intermediate IIa, 591 mg, 1Eq, 1.74mmol) of 3-hydroxy-2-(hydroxymethyl (9Z,12Z)-octadeca-9,12-dienoate (intermediate III, 640mg, 1Eq, 1.74mmol), DIPEA (673mg, 904μL, 3E q, 5.21 mmol), and DMAP (42.4 mg, 0.2 Eq, 347 μmol Finally, EDC (666 mg, 2 Eq, 3.47 mmol) was added, and 23 The mixture was stirred at 4° C. for 18 hours. After this time, the reaction mixture was concentrated and purified by flash column chromatography. HPLC (50 g silica, 0-40% ethyl acetate in hexane over 12 column volumes) The product was purified by 3-((4,4-bis(((Z)-oct-5-en-1-yl) )oxy)butanoyl)oxy)-2-(hydroxymethyl)propyl(9Z,12Z) -octadeca-9,12-dienoate (450 mg, 38%) was obtained as a colorless oil. . 1 H NMR (500 MHz, chloroform-d) δ 5.43–5.27 (m, 8H), 4.49 (t, J = 5.5 Hz, 1H), 4.23 - 4.12 (m, 4H), 3.65 - 3.60 (m, 2H), 3.58 (dt, J = 9.3, 6.6 Hz, 2H), 3 .41 (dt, J = 9.3, 6.6 Hz, 2H), 2.81 - 2.73 (m, 2H), 2.41 (t, J = 7.5 Hz, 2H), 2. 32 (dd, J = 7.9, 7.2 Hz, 2H), 2.25 - 2.15 (m, 2H), 2.12 - 1.97 (m, 12H), 1.94 (d dd, J = 8.0, 7.2, 5.5 Hz, 2H), 1.67 - 1.53 (m, 8H), 1.45 - 1.26 (m, 17H), 0.96 ( t, J = 7.6 Hz, 5H), 0.92 - 0.87 (m, 3H)

[0187] Step 5: 3-((4,4-bis(((Z)-oct-5-en-1-yl)oxy) )butanoyl)oxy)-2-((((3-(diethylamino)propoxy)carbonyl )oxy)methyl)propyl (9Z,12Z)-octadeca-9,12-dienoate ( Example 1)

[0188] [ka]

[0189] The following is representative of general procedure D: 3-((4, 4-bis(((Z)-oct-5-en-1-yl)oxy)butanoyl)oxy)-2 -(hydroxymethyl)propyl(9Z,12Z)-octadeca-9,12-dienoate A solution of 100 mg of intermediate IVa (1 Eq, 145 μmol) in pyridine (22. 9mg, 23.3μL, 2Eq, 289μmol), DMAP (4.42mg, 0.25 Eq, 36.2 μmol) and 4-nitrophenyl chloroformate (58.3 mg, 2 Eq The resulting mixture was stirred at 23° C. for 1 hour. Then, DIPEA (74.8 mg, 101 μL, 4 Eq, 579 μmol) and and 3-(diethylamino)propan-1-ol (76.0 mg, 85.9 μL, 4 Eq The resulting mixture was stirred at 23°C for an additional 18 hours. After this time, the reaction mixture was diluted with dichloromethane (10 mL) and diluted with 0.75 M sodium carbonate aqueous sodium chloride (10 mL), water (10 mL), and aqueous saturated sodium chloride (10 mL). The resulting organic layer was dried over sodium sulfate, concentrated, and the residue was Column chromatography (silica 10 g, 0-2% ethanol in dichloromethane over 12 min) 5% methanol). (3-(diethylamino)propan-1-yl)oxy)butanoyl)oxy)-2-( ... (9Z,12Z)-octadeca-9, The 12-dienoate (81 mg, 66%) was obtained as a pale yellow oil. 1 H NMR (500 MHz, Chloroform-d) δ 5.43 - 5.27 (m, 8H), 4.49 (t, J = 5.5 Hz, 1H), 4.22 - 4.11 ( m, 8H), 3.57 (dt, J = 9.3, 6.6 Hz, 2H), 3.41 (dt, J = 9.3, 6.6 Hz, 2H), 2.81 - 2 .74 (m, 2H), 2.51 (q, J = 7.1 Hz, 6H), 2.45 - 2.36 (m, 3H), 2.31 (dd, J = 8.0, 7 .2 Hz, 2H), 2.11 - 1.97 (m, 14H), 1.92 (ddd, J = 8.2, 7.1, 5.5 Hz, 2H), 1.86 - 1 .77 (m, 2H), 1.65 - 1.53 (m, 8H), 1.49 - 1.23 (m, 14H), 1.01 (t, J = 7.1 Hz, 6H) , 0.95 (t, J = 7.5 Hz, 6H), 0.92 - 0.86 (m, 3H). LCMS (Method A): Found m / z for (M+H) = 848.7, RT = 3.73 minutes.

[0190] [Example 2] 3-((4,4-bis(((Z)-oct-5-en-1-yl)oxy)butanoyl) oxy)-2-(((4-(pyrrolidin-1-yl)butanoyl)oxy)methyl)pro Pyr(9Z,12Z)-octadeca-9,12-dienoate

[0191] [ka]

[0192] The following is representative of general procedure E: 3-((4, 4-bis(((Z)-oct-5-en-1-yl)oxy)butanoyl)oxy)-2 -(hydroxymethyl)propyl(9Z,12Z)-octadeca-9,12-dienoate A mixture of 4-(pyrrolidinediamine)- ... (1-phenyl-1-yl)butanoic acid hydrochloride (26 mg, 1.2 Eq, 0.14 mmol, DIPEA (73 mg, 98 μL, 5 Eq, 0.56 mmol), and DMAP (2.8 mg, 0 Finally, EDC (43 mg, 2 Eq, 0.23 mmHg) was added. ol) was added and stirred at 23° C. for 18 hours. After this time, the reaction mixture was concentrated and Rush column chromatography (10 g silica, in DCM over 12 column volumes) 0-20% methanol). 5-en-1-yl)oxy)butanoyl)oxy)-2-(((4-(pyrrolidine-1 -yl)butanoyl)oxy)methyl)propyl(9Z,12Z)-octadeca-9,1 The 2-dienoate (74 mg, 78%) was obtained as a colorless oil. 1 H NMR (500 MHz, C12H-d) δ 5.42 - 5.28 (m, 8H), 4.49 (t, J = 5.6 Hz, 1H), 4.15 - 4.10 (m, 6H), 3.57 (dt, J = 9.3, 6.6 Hz, 2H), 3.41 (dt, J = 9.3, 6.6 Hz, 2H), 2.77 (tdq, J = 7.1, 1.4, 0.8 Hz, 2H), 2.57 - 2.43 (m, 6H), 2.43 - 2.35 (m, 5H), 2.30 (dd, J = 8.0, 7.2 Hz, 2H), 2.11 - 1.97 (m, 12H), 1.92 (ddd, J = 8.3, 7.2, 5.6 Hz, 2H), 1.89 - 1.73 (m, 6H), 1.68 - 1.52 (m, 8H), 1.48 - 1.24 (m, 16H), 0.95 (t, J = 7. 6 Hz, 6H), 0.92 - 0.85 (m, 3H). LCMS (Method A): (M+H) found m / z = 830.7, RT = 3.7 5 minutes

[0193] The following intermediates Ib-Ih were prepared according to general procedure A, but with the addition of the alcohol building block. Changed the block.

[0194] [ka]

[0195] Intermediate Ib, 4,4-bis((3,7-dimethyloct-6-en-1-yl)oxy) ) Butanenitrile: Prepared from rac-citronellol on a 9.5 mmol scale. Amount: 0.87g (24%).

[0196] [ka]

[0197] Intermediate Ic, 4,4-bis((4,4,5,5,5-pentafluoropentyl)oxy) ) Butanenitrile: 4,4,5,5,5-pentafluoropentane in 9.5 mmol scale Prepared from tan-1-ol. Yield 1.8g (45%).

[0198] [ka]

[0199] Intermediate Id, 4,4-bis(5,5,6,6,6-pentafluorohexyl)oxy Butanenitrile: 5,5,6,6,6-pentafluorohexanitrile on a 9.5 mmol scale Prepared from benzo-1-ol. Yield 0.70 g (16%).

[0200] [ka]

[0201] Intermediate Ie, 4,4-bis((4,4,5,5,6,6,7,7,7-nonafluorohe Butyl)oxy)butanenitrile: 4,4,5,5,6,6, on a 9.5 mmol scale Prepared from 7,7,7-nonafluoroheptan-1-ol. Yield: 2.0 g (34%) .

[0202] [ka]

[0203] Intermediate If, ​​4,4-bis((7,7,8,8,8-pentafluorooctyl)oxy) ) Butanenitrile: 7,7,8,8,8-pentafluorooctane in 9.5 mmol scale Prepared from tan-1-ol. Yield 1.6g (33%). 1 H NMR (500 MHz, chloroform) Lum-d) δ 4.57 (t, J = 5.3 Hz, 1H), 3.61 (dt, J = 9.2, 6.5 Hz, 2H), 3.45 (dt, J = 9.3, 6.5 Hz, 2H), 2.43 (t, J = 7.3 Hz, 2H), 2.09 - 1.91 (m, 6H), 1.65 - 1.53 (m, 8H), 1.47 - 1.35 (m, 8H).

[0204] [ka]

[0205] Intermediate Ig, 4,4-bis(4-(trifluoromethyl)phenethoxy)butannitrile 1. Prepared from (4-trifluoromethylphenyl)ethanol on a 9.5 mmol scale The yield was 2.2 g (52%).

[0206] [ka]

[0207] Intermediate Ih, 4,4-bis(3-cyclohexylpropoxy)butanenitrile: 9.5 Prepared on a mmol scale from 3-cyclohexylpropan-1-ol. Yield 2.3 g (68%).

[0208] The following intermediates IIb-IIh were prepared from the corresponding intermediates Ib-Ih according to general procedure B: Prepared.

[0209] [ka]

[0210] Intermediate IIb, 4,4-bis((3,7-dimethyloct-6-en-1-yl)oxa 4,4-bis((3,7-dimethylocta-6))butanoic acid: 2.3 mmol scale Prepared from (1-en-1-yl)oxy)butanenitrile (intermediate Ib). Yield 0.88 g(97%). 1 H NMR (500 MHz, chloroform-d) δ 5.09 (dddd, J = 7.1, 5.7, 2.9, 1.4 Hz, 2H), 4.51 (t, J = 5.5 Hz, 1H), 3.62 (dddd, J = 16.2, 14.0, 6.9, 1.5 Hz, 2H), 3.45 (ddt, J = 16.7, 9.2, 6.7 Hz, 2H), 2.45 (t, J = 7.3 Hz, 2H), 2.06 - 1. 89 (m, 6H), 1.68 (d, J = 1.4 Hz, 6H), 1.66 - 1.52 (m, 8H), 1.42 - 1.29 (m, 5H), 1.16 (dddd, J = 13.4, 9.8, 7.8, 5.8 Hz, 3H), 0.89 (dd, J = 6.6, 1.7 Hz, 6H).

[0211] [ka]

[0212] Intermediate IIc, 4,4-bis((4,4,5,5,5-pentafluoropentyl)oxy) 4,4-bis((4,4,5,5,5-penta)-3-butanoic acid: 4.3 mmol scale Prepared from (fluoropentyl)oxy)butanenitrile (Intermediate Ic). Yield: 1.6 g (83%). 1 H NMR (500 MHz, chloroform-d) δ 4.55 (t, J = 5.4 Hz, 1H), 3.65 ( dt, J = 9.5, 6.1 Hz, 2H), 3.49 (dt, J = 9.5, 6.1 Hz, 2H), 2.43 (t, J = 7.2 Hz, 2 H), 2.22 - 2.06 (m, 4H), 1.92 - 1.82 (m, 6H).

[0213] [ka]

[0214] Intermediate IId, 4,4-bis((5,5,6,6,6-pentafluorohexyl)oxy) 4,4-bis((5,5,6,6,6-penta)-3-butanoic acid: 1.6 mmol scale Prepared from (fluorohexyl)oxy)butanenitrile (Intermediate Id). Yield 0.53 g (73%).

[0215] [ka]

[0216] Intermediate IIe, 4,4-bis((4,4,5,5,6,6,7,7,7-nonafluoro) Heptyl)oxy)butanoic acid: 4,4-bis((4,4,5, 5,6,6,7,7,7-Nonafluoroheptyl)oxy)butanenitrile (Intermediate Ie ) Yield: 1.9 g (91%). 1 H NMR (500 MHz, chloroform-d) δ 4.55 (t, J = 5.5 Hz, 1H), 3.65 (dt, J = 9.5, 6.1 Hz, 2H), 3.50 (dt, J = 9.5, 6.1 Hz, 2H), 2.43 (td, J = 7.2, 3.5 Hz, 2H), 2.18 (tt, J = 18.5, 7.8 Hz, 4H), 2.02 - 1. 83 (m, 6H).

[0217] [ka]

[0218] Intermediate IIf, 4,4-bis((7,7,8,8,8-pentafluorooctyl)oxa 4,4-bis((7,7,8,8,8-penta)-3,4-dibutanoic acid: 3.1 mmol scale Prepared from (fluorooctyl)oxy)butanenitrile (Intermediate If). Yield: 1.6 g (96%). 1 H NMR (500 MHz, chloroform-d) δ 4.50 (t, J = 5.5 Hz, 1H), 3.57 ( dt, J = 9.4, 6.6 Hz, 2H), 3.41 (dt, J = 9.4, 6.6 Hz, 2H), 2.38 (t, J = 7.5 Hz, 2 H), 2.08 - 1.87 (m, 6H), 1.69 - 1.51 (m, 8H), 1.41 - 1.38 (m, J = 3.5 Hz, 8H).

[0219] [ka]

[0220] Intermediate IIg, 4,4-bis(4-(trifluoromethyl)phenethoxy)butanoic acid: 4,4-bis(4-(trifluoromethyl)phenethoxy)butanol on a 5.0 mmol scale Prepared from ethanenitrile (intermediate Ig). Yield 1.7 g (74%). 1 H NMR (500 MHz, Chloroform-d) δ 7.52 (d, J = 7.6 Hz, 4H) 7.27 (d, J = 7.6 Hz, 4H), 4.49 (t, J = 5.5 Hz, 1H), 3.66 (dt, J = 9.4, 6.6 Hz, 2H), 3.53 (dt, J = 9.4, 6.6 Hz, 2H), 2.84 (t, J = 6.6 Hz, 4H), 2.31 (t, J = 7.3 Hz, 2H), 1.88 (td, J = 7.4, 5.6 Hz, 2H).

[0221] [ka]

[0222] Intermediate IIh, 4,4-bis(3-cyclohexylpropoxy)butanoic acid: 6.5 mm 4,4-bis(3-cyclohexylpropoxy)butanenitrile (intermediate) Ih). Yield 1.9 g (79%). 1 H NMR (500 MHz, chloroform-d) δ 4.50 (t, J = 5.5 Hz, 1H), 3.55 (dt, J = 9.3, 6.8 Hz, 2H), 3.40 (dt, J = 9.3, 6.8 Hz, 2H), 2.41 (t, J = 7.3 Hz, 2H), 1.92 (td, J = 7.4, 5.4 Hz, 2H), 1.76 - 1.49 (m, 14H), 1.33 - 1.06 (m, 12H), 0.94 - 0.73 (m, 4H).

[0223] The following intermediates IVb to IVh were synthesized from the corresponding intermediates IIb to IIh and intermediate III. It was prepared according to general procedure C from

[0224] [ka]

[0225] Intermediate IVb, 3-((4,4-bis((3,7-dimethyloct-6-en-1-yl) 2-(hydroxymethyl)propyl (9Z, 12Z)- ... )-octadeca-9,12-dienoate: 4,4-bis(( From 3,7-dimethyloct-6-en-1-yl)oxy)butanoic acid (intermediate IIb) Prepared in a yield of 0.37g (45%). 1 H NMR (500 MHz, chloroform-d) δ 5.43 - 5 .28 (m, 4H), 5.13 - 5.06 (m, 2H), 4.49 (t, J = 5.5 Hz, 1H), 4.23 - 4.12 (m, 4H), 3.68 - 3.55 (m, 4H), 3.50 - 3.38 (m, 2H), 2.80 - 2.74 (m, 2H), 2.41 (t, J = 7.5 Hz, 2H), 2.35 - 2.29 (m, 2H), 2.25 - 2.15 (m, 2H), 2.10 - 1.89 (m, 10H), 1.74 - 1.66 (m, 6H), 1.66 - 1.51 (m, 12H), 1.44 - 1.23 (m, 18H), 1.22 - 1.11 (m, 2H), 0.96 - 0.84 (m, 9H).

[0226] [ka]

[0227] Intermediate IVc, 3-((4,4-bis((4,4,5,5,5-pentafluoropentyl) 2-(hydroxymethyl)propyl (9Z, 12Z)- ... )-octadeca-9,12-dienoate: 4,4-bis(( From 4,4,5,5,5-pentafluoropentyl)oxy)butanoic acid (intermediate IIc) Prepared in the following yield: 0.40 g (37%). 1 H NMR (500 MHz, chloroform-d) δ 5.43 - 5 .28 (m, 4H), 4.53 (t, J = 5.5 Hz, 1H), 4.23 - 4.08 (m, 4H), 3.68 - 3.57 (m, 4H), 3.49 (dt, J = 9.5, 6.0 Hz, 2H), 2.80 - 2.74 (m, 2H), 2.40 (t, J = 7.4 Hz, 2H), 2.32 (dd, J = 7.9, 7.2 Hz, 2H), 2.23 - 2.00 (m, 11H), 1.99 - 1.81 (m, 6H), 1.68 - 1.55 (m, 4H), 1.40 - 1.24 (m, 11H), 0.91 - 0.86 (m, 3H).

[0228] [ka]

[0229] Intermediate IVd, 3-((4,4-bis((5,5,6,6,6-pentafluorohexyl) 2-(hydroxymethyl)propyl (9Z, 12Z)- ... )-octadeca-9,12-dienoate: 4,4-bis(( From 5,5,6,6,6-pentafluorohexyl)oxy)butanoic acid (intermediate IId) Prepared in the following yield: 0.20 g (23%).

[0230] [ka]

[0231] Intermediate IVe, 3-((4,4-bis((4,4,5,5,6,6,7,7,7-nonano) Fluoroheptyloxybutanoyloxy-2-(hydroxymethyl)propyl (9Z,12Z)-octadeca-9,12-dienoate: 4 on a 1.4 mmol scale ,4-bis((4,4,5,5,6,6,7,7,7-nonafluoroheptyl)oxy) Prepared from butanoic acid (intermediate IIe). Yield 0.50 g (37%). 1 H NMR (500 MHz, Chloroform-d) δ 5.43 - 5.28 (m, 4H), 4.54 (t, J = 5.5 Hz, 1H), 4.24 - 4.08 ( m, 4H), 3.68 - 3.60 (m, 4H), 3.51 (dt, J = 9.5, 6.1 Hz, 2H), 2.80 - 2.74 (m, 2H) , 2.41 (t, J = 7.4 Hz, 2H), 2.32 (dd, J = 8.0, 7.2 Hz, 2H), 2.26 - 2.11 (m, 5H), 2.10 - 2.01 (m, 5H), 1.99 - 1.84 (m, 6H), 1.65 - 1.59 (m, 2H), 1.57 (s, 2H), 1. 40 - 1.23 (m, 12H), 0.92 - 0.86 (m, 3H).

[0232] [ka]

[0233] Intermediate IVf, 3-((4,4-bis((7,7,8,8,8-pentafluorooctyl) 2-(hydroxymethyl)propyl (9Z, 12Z)- ... )-octadeca-9,12-dienoate: 4,4-bis(( From 7,7,8,8,8-pentafluorooctyl)oxy)butanoic acid (intermediate IIf) Prepared in the following yield: 0.39g (41%). 1 H NMR (500 MHz, chloroform-d) δ 5.43 - 5 .28 (m, 4H), 4.49 (t, J = 5.5 Hz, 1H), 4.23 - 4.12 (m, 4H), 3.62 (t, J = 5.9 Hz, 2H), 3.57 (dt, J = 9.3, 6.5 Hz, 2H), 3.41 (dt, J = 9.3, 6.6 Hz, 2H), 2.81 - 2.7 4 (m, 2H), 2.41 (t, J = 7.5 Hz, 2H), 2.32 (dd, J = 7.9, 7.2 Hz, 2H), 2.24 - 2.16 (m, 2H), 2.10 - 1.89 (m, 12H), 1.66 - 1.57 (m, 9H), 1.46 - 1.24 (m, 21H), 0.92 - 0.86 (m, 3H).

[0234] [ka]

[0235] Intermediate IVg, 3-((4,4-bis(4-(trifluoromethyl)phenethoxy)butoxy)- (9Z,12Z)-octadeca(2-hydroxymethyl)propyl 2-( ... -9,12-dienoate: 4,4-bis(4-(trifluoromethyl)-4,4-difluoromethyl ... Prepared from (trimethyl)phenethoxy)butanoic acid (Intermediate IIg). Yield: 0.38 g (3 5%). 1 H NMR (500 MHz, chloroform-d) δ 7.54 - 7.50 (m, 4H), 7.29 - 7.26 (m, 4H), 5.43 - 5.28 (m, 4H), 4.48 (t, J = 5.5 Hz, 1H), 4.22 - 4.06 (m, 5H), 3.69 - 3.62 (m, 2H), 3.60 (t, J = 5.7 Hz, 2H), 3.57 - 3.47 (m, 2H), 2.85 (t, J = 6.7 H z, 4H), 2.80 - 2.74 (m, 2H), 2.38 - 2.23 (m, 4H), 2.23 - 2.12 (m, 2H), 2.10 - 2. 01 (m, 4H), 1.95 - 1.85 (m, 2H), 1.65 - 1.56 (m, 4H), 1.41 - 1.23 (m, 11H), 0.92 - 0.86 (m, 3H).

[0236] [ka]

[0237] Intermediate IVh, 3-((4,4-bis(3-cyclohexylpropoxy)butanoyl) Oxy)-2-(hydroxymethyl)propyl(9Z,12Z)-octadeca-9,12 -Dienoate: 4,4-bis(3-cyclohexylpropoxy) Prepared from butanoic acid (intermediate IIh). Yield: 0.40 g (41%). 1 H NMR (500 MHz, chloroform-d) δ 5.43 - 5.28 (m, 4H), 4.49 (t, J = 5.5 Hz, 1H), 4.23 - 4. 11 (m, 4H), 3.62 (t, J = 5.8 Hz, 2H), 3.55 (dt, J = 9.3, 6.8 Hz, 2H), 3.39 (dt, J = 9.3, 6.8 Hz, 2H), 2.77 (ttd, J = 7.0, 1.4, 0.7 Hz, 2H), 2.41 (t, J = 7.5 Hz, 2H), 2.32 (dd, J = 7.9, 7.2 Hz, 2H), 2.25 - 2.16 (m, 1H), 2.09 - 2.01 (m, 4H), 1.93 (ddd, J = 7.9, 7.2, 5.5 Hz, 2H), 1.74 - 1.50 (m, 18H), 1.42 - 1.09 (m, 25H) , 0.92 - 0.83 (m, 7H).

[0238] The following Examples 3-11 were prepared according to general procedure D from intermediate IVa.

[0239] [ka]

[0240] [Example 3] 3-((4,4-bis(((Z)-oct-5-en-1-yl)oxy)butanoyl) oxy)-2-(((((1-ethylpiperidin-3-yl)methoxy)carbonyl)o Oxy)methyl)propyl (9Z,12Z)-octadeca-9,12-dienoate: 0. Intermediate IVa and (1-ethylpiperidin-3-yl)methanoic acid were synthesized on a 15 mmol scale. Prepared from ethanol. Yield: 0.082 g (66%). 1 H NMR (500 MHz, chloroform-d) δ 5.43 - 5.27 (m, 8H), 4.49 (t, J = 5.6 Hz, 1H), 4.22 - 4.12 (m, 6H), 4.09 - 3. 93 (m, 1H), 3.57 (dt, J = 9.3, 6.6 Hz, 2H), 3.41 (dt, J = 9.4, 6.6 Hz, 2H), 2.80 - 2.74 (m, 2H), 2.47 - 2.36 (m, 4H), 2.31 (dd, J = 8.0, 7.2 Hz, 2H), 2.12 - 1.9 6 (m, 12H), 1.96 - 1.85 (m, 3H), 1.82 - 1.67 (m, 3H), 1.67 - 1.51 (m, 10H), 1.48 - 1.24 (m, 21H), 1.08 (t, J = 7.2 Hz, 3H), 0.96 (t, J = 7.6 Hz, 6H), 0.92 - 0.8 5 (m, 3H). LCMS (Method A): (M+H) found m / z = 860.6, RT = 3.75 min.

[0241] [ka]

[0242] [Example 4] 3-((4,4-Bis(((Z)-oct-5-en-1-yl)oxy)butanoyl) oxy)-2-((((3-(dipropylamino)propoxy)carbonyl)oxy)meth yl)propyl (9Z,12Z)-octadeca-9,12-dienoate: 0.072 m mol scale from intermediate IVa and 3-(dipropylamino)propan-1-ol Prepared. Yield 0.057 g (90%). LCMS (Method A): Measured value of (M+H) m / z = 876.6, RT = 3.76 min.

[0243] [Chemical formula]

[0244] [Example 5] 3-((4,4-Bis(((Z)-oct-5-en-1-yl)oxy)butanoyl) oxy)-2-((((2-(1-methylpyrrolidin-2-yl)ethoxy)carbonyl )oxy)methyl)propyl (9Z,12Z)-octadeca-9,12-dienoate: 0.072 mmol scale from intermediate IVa and 2-(1-methylpyrrolidin-2-i l)ethane-1-ol. Yield 0.042 g (69%). LCMS (Method A): (M +H) measured value m / z = 846.7, RT = 3.72 min.

[0245] [Chemical formula]

[0246] ​​​ oxy)-2-(((((1-ethylpyrrolidin-3-yl)methoxy)carbonyl)o Oxy)methyl)propyl (9Z,12Z)-octadeca-9,12-dienoate: 0. Intermediate IVa and (1-ethylpyrrolidin-3-yl)methanone were synthesized on a 0.72 mmol scale. Prepared from alcohol. Yield 0.039 g (64%). LCMS (Method A): Found m / z for (M+H) = 846.6, RT = 3.72 minutes.

[0247] [ka]

[0248] [Example 7] 3-((4,4-bis(((Z)-oct-5-en-1-yl)oxy)butanoyl) oxy)-2-((((3-(diethylamino)-2-hydroxypropoxy)carbonyl (9Z,12Z)-octadeca-9,12-dienoate : Intermediate IVa and 3-(diethylamino)propane- Prepared from 1,2-diol. Yield 0.002 g (3%). LCMS (Method A): (M+H) Measured m / z = 864.6, RT = 3.66 minutes.

[0249] [ka]

[0250] [Example 8] 3-((4,4-bis(((Z)-oct-5-en-1-yl)oxy)butanoyl) oxy)-2-((((3-(dimethylamino)propoxy)carbonyl)oxy)methyl (9Z,12Z)-octadeca-9,12-dienoate: 0.077mm Prepared on a 1 / 2 scale from intermediate IVa and 3-(dimethylamino)propan-1-ol Yield 0.022 g (35%). LCMS (Method A): (M+H) found m / z = 820.7, RT = 3.77 minutes.

[0251] [ka]

[0252] [Example 9] 3-((4,4-bis(((Z)-oct-5-en-1-yl)oxy)butanoyl) oxy)-2-((((3-(pyrrolidin-1-yl)propoxy)carbonyl)oxy )Methyl)propyl (9Z,12Z)-octadeca-9,12-dienoate: 0.07 Intermediate IVa and 3-(pyrrolidin-1-yl)propan-1-yl were synthesized on a 7 mmol scale. Prepared from ethanol. Yield 0.027 g (41%). LCMS (Method A): Found m / z for (M+H) = 846.7, RT = 3.76 minutes.

[0253] [ka]

[0254] [Example 10] 3-((4,4-bis(((Z)-oct-5-en-1-yl)oxy)butanoyl) oxy)-2-((((2-(1-methylpiperidin-2-yl)ethoxy)carbonyl )Oxy)methyl)propyl (9Z,12Z)-octadeca-9,12-dienoate: Intermediate IVa and 2-(1-methylpiperidin-2-yl)- Prepared from (Methyl)ethan-1-ol. Yield 0.039 g (52%). LCMS (Method A): +H) Found m / z = 860.6, RT = 3.83 min.

[0255] [ka]

[0256] [Example 11] 3-(((3-(azetidin-1-yl)propoxy)carbonyl)oxy)-2-(( (4,4-bis(((Z)-oct-5-en-1-yl)oxy)butanoyl)oxy )Methyl)propyl (9Z,12Z)-octadeca-9,12-dienoate: 0.08 Intermediate IVa and 3-(azetidin-1-yl)propan-1-yl were synthesized on a 7 mmol scale. Prepared from ethanol. Yield 0.041 g (57%). LCMS (Method A): Found m / z for (M+H) = 832.5, RT = 3.74 minutes.

[0257] The following Examples 12 and 13 were prepared from intermediate IVa according to general procedure E.

[0258] [ka]

[0259] [Example 12] 3-((4,4-bis(((Z)-oct-5-en-1-yl)oxy)butanoyl) oxy)-2-(((3-(4-methylpiperazin-1-yl)propanoyl)oxy) Methyl)propyl (9Z,12Z)-octadeca-9,12-dienoate: 0.072 Intermediate IVa and 3-(4-methylpiperazin-1-yl)propanol were synthesized on a mmol scale. Prepared from phosphonic acid dihydrochloride. Yield 0.056g (91%). 1 H NMR (500 MHz, chloroform) Lum-d) δ 5.42 - 5.27 (m, 8H), 4.49 (t, J = 5.6 Hz, 1H), 4.21 - 4.10 (m, 6H), 3 .64 - 3.53 (m, 2H), 3.41 (dt, J = 9.3, 6.6 Hz, 2H), 2.80 - 2.74 (m, 2H), 2.72 - 2.65 (m, 2H), 2.62 - 2.46 (m, 6H), 2.46 - 2.35 (m, 7H), 2.35 - 2.23 (m, 4H), 2.1 2 - 1.96 (m, 13H), 1.96 - 1.89 (m, 2H), 1.67 - 1.52 (m, 9H), 1.48 - 1.23 (m, 15H) ), 0.95 (t, J = 7.6 Hz, 6H), 0.92 - 0.86 (m, 3H). LCMS (Method A): Found m / z for (M+H) = 845.6, RT = 3.67 minutes.

[0260] [ka]

[0261] [Example 13] 3-((4,4-bis(((Z)-oct-5-en-1-yl)oxy)butanoyl) oxy)-2-((((9Z,12Z)-octadeca-9,12-dienoyl)oxy) Methyl)propyl 1,3-dimethylpyrrolidine-3-carboxylate: 0.077mm Prepared on a 1 / 2 scale from intermediate IVa and 1,3-dimethylpyrrolidine-3-carboxylic acid Yield 0.007 g (11%). LCMS (Method A): (M+H) found m / z = 816.6, RT = 3.76 minutes.

[0262] The following examples 14-22 were prepared according to general procedure D from intermediate IVb.

[0263] [ka]

[0264] [Example 14] 3-((4,4-bis((3,7-dimethyloct-6-en-1-yl)oxy)buta (((3-(diethylamino)propoxy)carbonyl)oxy)-2-((((3-(diethylamino)propoxy)carbonyl)oxy) methyl)propyl (9Z,12Z)-octadeca-9,12-dienoate: 0.1 Intermediate IVb and 3-(diethylamino)propan-1-ol on a 3 mmol scale Prepared from. Yield 0.056g (46%). 1 H NMR (500 MHz, chloroform-d) δ 5. 43 - 5.28 (m, 4H), 5.13 - 5.06 (m, 2H), 4.48 (t, J = 5.5 Hz, 1H), 4.22 - 4.10 (m , 8H), 3.66 - 3.55 (m, 2H), 3.50 - 3.38 (m, 2H), 2.80 - 2.74 (m, 2H), 2.51 (q, J = 7.1 Hz, 6H), 2.48 - 2.37 (m, 3H), 2.31 (dd, J = 8.0, 7.2 Hz, 2H), 2.05 (dd, J = 7.2, 1.1 Hz, 3H), 2.02 - 1.89 (m, 3H), 1.81 (p, J = 6.7 Hz, 2H), 1.73 - 1.50 (m, 23H), 1.45 - 1.23 (m, 17H), 1.22 - 1.11 (m, 2H), 1.01 (t, J = 7.1 Hz, 6H), 0 0.95 - 0.82 (m, 9H). LCMS (Method A): (M+H) found m / z = 904.8, RT = 4.06 min.

[0265] [ka]

[0266] [Example 15] 3-((4,4-bis((3,7-dimethyloct-6-en-1-yl)oxy)buta ((((1-ethylpiperidin-3-yl)methoxy)carbo (9Z,12Z)-octadeca-9,12-dienoic acid Intermediate IVb and (1-ethylpiperidin-3-yl) ) Prepared from methanol. Yield 0.070 g (57%). LCMS (Method A): Observed (M+H) Value m / z = 916.7, RT = 4.06 min.

[0267] [ka]

[0268] [Example 16] 3-((4,4-bis((3,7-dimethyloct-6-en-1-yl)oxy)buta 2-(((((1-ethylpyrrolidin-3-yl)methoxy)carbo (9Z,12Z)-octadeca-9,12-dienoic acid IVb and (1-ethylpyrrolidin-3-yl) ) Prepared from methanol. Yield 0.040 g (55%). LCMS (Method A): Observed (M+H) Value m / z = 902.7, RT = 4.00 min.

[0269] [ka]

[0270] [Example 17] 3-((4,4-bis((3,7-dimethyloct-6-en-1-yl)oxy)buta (2-(1-methylpiperidin-3-yl)ethoxy)-2-((((2-(1-methylpiperidin-3-yl)ethoxy) (9Z,12Z)-octadeca-9,12-dieno ester: intermediate IVb and 2-(1-methylpiperidine- 3-yl)ethan-1-ol. Yield 0.038 g (52%). LCMS (Method A): (M+H) Found m / z = 916.8, RT = 3.94 min.

[0271] [ka]

[0272] [Example 18] 3-((4,4-bis((3,7-dimethyloct-6-en-1-yl)oxy)buta 4-morpholinobutoxy)carbonyl)oxy)methyl ) Propyl (9Z,12Z)-octadeca-9,12-dienoate: 0.08 mmol This was prepared on a large scale from intermediate IVb and 4-morpholinobutan-1-ol. Yield: 0 0.039g (52%). LCMS (Method A): (M+H) found m / z = 932.6, RT = 3.95 min.

[0273] [ka]

[0274] [Example 19] 3-((4,4-bis((3,7-dimethyloct-6-en-1-yl)oxy)buta 1r,4r)-4-morpholinocyclohexyl)oxy)-2-((((((1r,4r)-4-morpholinocyclohexyl)oxy) Oxy)carbonyl)oxy)methyl)propyl(9Z,12Z)-octadeca-9,12 -dienoate: intermediate IVb and trans-4-mol on a 0.08 mmol scale Prepared from cyclohexyl 1-ol. Yield 0.036g (47%). LCMS ( Method A): (M+H) found m / z = 958.6, RT = 3.95 min.

[0275] [ka]

[0276] [Example 20] 3-((4,4-bis((3,7-dimethyloct-6-en-1-yl)oxy)buta (4-(4-methylpiperazin-1-yl)butoxy)-2-((((4-methylpiperazin-1-yl)butoxy) (9Z,12Z)-octadeca-9,12-dieno 4-(4-methylpiperazine-)-4-methylpiperazine ... 1-yl)butan-1-ol. Yield 0.027g (36%). LCMS (Method A): (M+H) Found m / z = 945.7, RT = 3.95 min.

[0277] [ka]

[0278] [Example 21] 3-((4,4-bis((3,7-dimethyloct-6-en-1-yl)oxy)buta (((((1R,3s,5S)-8-methyl-8-azabicyclo[2.2.1.2.2.1]methyl)oxy)-2-( ...1R,3s,5S)-8-methyl-8-azabicyclo[2.2.1.2.1]methyl)oxy (ii)[3.2.1]octan-3-yl)oxy)carbonyl)oxy)methyl)propyl (9Z,12Z)-octadeca-9,12-dienoate: on a 0.08 mmol scale Prepared from intermediate IVb and pseudotropine. Yield 0.019 g (26%). LCMS (Method A): (M+H) found m / z = 914.6, RT = 3.88 min.

[0279] [ka]

[0280] [Example 22] 3-((4,4-bis((3,7-dimethyloct-6-en-1-yl)oxy)buta benzyl)oxy)-2-(((((2-((dimethylamino)methyl)benzyl)oxy )carbonyl)oxy)methyl)propyl(9Z,12Z)-octadeca-9,12-di Enoate: Intermediate IVb and (2-((dimethylamino) (methyl)phenyl)methanol. Yield 0.030 g (40%). LCMS (method Method A): (M+H) measured m / z = 938.8, RT = 4.28 min.

[0281] The following Examples 23-30 were prepared according to general procedure E from intermediate IVb.

[0282] [ka]

[0283] [Example 23] 3-((4,4-bis((3,7-dimethyloct-6-en-1-yl)oxy)buta 4-(pyrrolidin-1-yl)butanoyl)oxy)methyl (9Z,12Z)-octadeca-9,12-dienoate: 0.09mmol Prepared on a full scale from intermediate IVb and 4-(pyrrolidin-1-yl)butanoic acid hydrochloride Yield 0.052 g (64%). LCMS (Method A): (M+H) found m / z = 886.7, RT = 3 .98 minutes.

[0284] [ka]

[0285] [Example 24] 3-((4,4-bis((3,7-dimethyloct-6-en-1-yl)oxy)buta 3-(4-methylpiperazin-1-yl)propanoyl)-2-(((3-(4-methylpiperazin-1-yl)propanoyl)oxy)-2-(((3-(4-methylpiperazin-1-yl)propanoyl) Oxy)methyl)propyl (9Z,12Z)-octadeca-9,12-dienoate: 0 Intermediate IVb and 3-(4-methylpiperazin-1-yl) Prepared from propanoic acid dihydrochloride. Yield 0.060 g (72%). LCMS (Method A): (M+H) Found m / z = 901.8, RT = 4.08 min.

[0286] [ka]

[0287] [Example 25] 3-((4,4-bis((3,7-dimethyloct-6-en-1-yl)oxy)buta 9Z,12Z)-octadeca-9,12-dienoyl)oxy)-2-((((9Z,12Z)-octadeca-9,12-dienoyl) Oxy)methyl)propyl 1'-ethyl-[1,4'-bipiperidine]-4-carboxy Rate: Intermediate IVb and 1'-ethyl-[1,4'-biphenyl] Prepared from piperidine-4-carboxylic acid dihydrochloride. Yield 0.021 g (27%). LC MS (Method A): (M+H) found m / z = 969.8, RT = 3.60 min.

[0288] [ka]

[0289] [Example 26] 3-((4,4-bis((3,7-dimethyloct-6-en-1-yl)oxy)buta 9Z,12Z)-octadeca-9,12-dienoyl)oxy)-2-((((9Z,12Z)-octadeca-9,12-dienoyl) Oxy)methyl)propyl 1-(pyridin-4-yl)piperidine-4-carboxylate Intermediate IVb and 1-(pyridin-4-yl)piperidine were synthesized on a 0.08 mmol scale. Prepared from azine-4-carboxylic acid. Yield 0.020 g (27%). LCMS (Method A): (M+H ) observed m / z = 935.5, RT = 3.88 min.

[0290] [ka]

[0291] [Example 27] 3-((4,4-bis((3,7-dimethyloct-6-en-1-yl)oxy)buta 5-(dimethylamino)pentanoyl)oxy)-2-(((5-(dimethylamino)pentanoyl)oxy)methyl) Propyl (9Z,12Z)-octadeca-9,12-dienoate: 0.08 mmol It was prepared in vacuo from intermediate IVb and 5-(dimethylamino)pentanoic acid hydrochloride. Amount 0.057 g (81%). LCMS (Method A): (M+H) found m / z = 874.8, RT = 3.95 min.

[0292] [ka]

[0293] [Example 28] 3-((4,4-bis((3,7-dimethyloct-6-en-1-yl)oxy)buta 4-(dipropylamino)butanoyl)oxy)-2-(((4-(dipropylamino)butanoyl)oxy)methyl) Propyl (9Z,12Z)-octadeca-9,12-dienoate: 0.08 mmol It was prepared in situ from intermediate IVb and 4-(dipropylamino)butanoic acid hydrochloride. Amount 0.062 g (84%). LCMS (Method A): (M+H) found m / z = 916.8, RT = 4.00 min.

[0294] [ka]

[0295] [Example 29] 3-((4,4-bis((3,7-dimethyloct-6-en-1-yl)oxy)buta 5-morpholinopentanoyl)oxy)-2-(((5-morpholinopentanoyl)oxy)methyl)propyl (9Z,12Z)-octadeca-9,12-dienoate: on a 0.08 mmol scale Prepared from intermediate IVb and 5-morpholinopentanoic acid hydrochloride. Yield 0.064 g ( 87%). LCMS (Method A): (M+H) found m / z = 916.8, RT = 3.96 min.

[0296] [ka]

[0297] [Example 30] 3-((4,4-bis((3,7-dimethyloct-6-en-1-yl)oxy)buta 2-((2-(1-methylpiperidin-4-yl)acetoxy)methyl)- ... (9Z,12Z)-octadeca-9,12-dienoate: 0.08mmol Prepared on a full scale from intermediate IVb and 2-(1-methylpiperidin-4-yl)acetic acid Yield 0.040 g (56%). LCMS (Method A): (M+H) found m / z = 886.7, RT = 3 .96 minutes.

[0298] The following Examples 31 and 32 were prepared according to general procedure D from intermediate IVc.

[0299] [ka]

[0300] [Example 31] 3-((4,4-bis((4,4,5,5,5-pentafluoropentyl)oxy)buta (((3-(diethylamino)propoxy)carbonyl)oxy)-2-((((3-(diethylamino)propoxy)carbonyl)oxy) methyl)propyl (9Z,12Z)-octadeca-9,12-dienoate: 0.1 Intermediate IVc and 3-(diethylamino)propan-1-ol on a 3 mmol scale Prepared from. Yield 0.042 g (35%). LCMS (Method A): Found m / z for (M+H) = 948.5 , RT = 3.49 minutes.

[0301] [ka]

[0302] [Example 32] 3-((4,4-bis((4,4,5,5,5-pentafluoropentyl)oxy)buta ((((1-ethylpiperidin-3-yl)methoxy)carbo (9Z,12Z)-octadeca-9,12-dienoic acid Intermediate IVc and (1-ethylpiperidin-3-yl) ) Prepared from methanol. Yield 0.050 g (41%). LCMS (Method A): Observed (M+H) Value m / z = 960.6, RT = 3.52 min.

[0303] The following Examples 33 and 34 were prepared according to general procedure D from intermediate IVd.

[0304] [ka]

[0305] [Example 33] 3-((4,4-bis((5,5,6,6,6-pentafluorohexyl)oxy)buta (((3-(diethylamino)propoxy)carbonyl)oxy)-2-((((3-(diethylamino)propoxy)carbonyl)oxy) methyl)propyl (9Z,12Z)-octadeca-9,12-dienoate: 0.1 Intermediate IVd and 3-(diethylamino)propan-1-ol on a 2 mmol scale Prepared from. Yield 0.044g (37%). 1 H NMR (500 MHz, chloroform-d) δ 5. 43 - 5.29 (m, 4H), 4.50 (t, J = 5.5 Hz, 1H), 4.22 - 4.09 (m, 8H), 3.60 (dt, J = 9.2, 5.8 Hz, 2H), 3.44 (dt, J = 9.3, 5.8 Hz, 2H), 2.77 (t, J = 6.7 Hz, 2H), 2.51 (q, J = 7.2 Hz, 6H), 2.46 - 2.37 (m, 3H), 2.31 (t, J = 7.6 Hz, 2H), 2.13 - 1.99 (m, 9H), 1.93 (td, J = 7.5, 5.4 Hz, 2H), 1.86 - 1.76 (m, 2H), 1.73 - 1.56 (m, 1 1H), 1.41 - 1.24 (m, 12H), 1.01 (t, J = 7.1 Hz, 6H), 0.92 - 0.83 (m, 3H). LCMS ( Method A): (M+H) found m / z = 976.6, RT = 3.52 min.

[0306] [ka]

[0307] [Example 34] 3-((4,4-bis((5,5,6,6,6-pentafluorohexyl)oxy)buta ((((1-ethylpiperidin-3-yl)methoxy)carbo (9Z,12Z)-octadeca-9,12-dienoic acid Intermediate IVd and (1-ethylpiperidin-3-yl) ) Prepared from methanol. Yield 0.035g (29%). 1 H NMR (500 MHz, chloroform) Lum-d) δ 5.43 - 5.29 (m, 4H), 4.51 (t, J = 5.5 Hz, 1H), 4.21 - 4.11 (m, 6H), 4 .05 (dd, J = 10.6, 5.8 Hz, 1H), 3.96 (dd, J = 10.6, 7.2 Hz, 1H), 3.60 (dt, J = 9 .3, 5.8 Hz, 2H), 3.44 (dt, J = 9.3, 5.8 Hz, 2H), 2.89 (dd, J = 35.3, 11.1 Hz, 2H) ), 2.77 (t, J = 6.7 Hz, 2H), 2.47 - 2.36 (m, 5H), 2.31 (t, J = 7.6 Hz, 2H), 2.13 - 1.83 (m, 9H), 1.78 - 1.55 (m, 15H), 1.42 - 1.23 (m, 17H), 1.08 (t, J = 7.2 Hz , 3H), 0.89 (t, J = 6.9 Hz, 3H). LCMS (Method A): (M+H) found m / z = 988.4, RT = 3. 52 minutes.

[0308] The following Examples 35 and 36 were prepared according to general procedure D from intermediate IVe.

[0309] [ka]

[0310] [Example 35] 3-((4,4-bis((4,4,5,5,6,6,7,7,7-nonafluoroheptyl )oxy)butanoyl)oxy)-2-((((3-(diethylamino)propoxy)carbonyl (9Z,12Z)-octadeca-9,12-dieno ate: intermediate IVe and 3-(diethylamino)propionate on a 0.12 mmol scale Prepared from benzo-1-ol. Yield 0.050 g (36%). LCMS (Method A): (M+H) Measured m / z = 1148.4, RT = 3.72 minutes.

[0311] [ka]

[0312] [Example 36] 3-((4,4-bis((4,4,5,5,6,6,7,7,7-nonafluoroheptyl )oxy)butanoyl)oxy)-2-(((((1-ethylpiperidin-3-yl)methyl (9Z,12Z)-octadeca-9,1 2-Dienoate: Intermediate IVe and (1-ethylpiperidine) were prepared on a 0.12 mmol scale. Prepared from (di-(3-methyl-3-yl)methanol. Yield 0.072 g (51%). LCMS (Method A) : (M+H) Found m / z = 1160.4, RT = 3.73 min.

[0313] The following examples 37-45 were prepared according to general procedure D from intermediate IVf.

[0314] [ka]

[0315] [Example 37] 3-((4,4-bis((7,7,8,8,8-pentafluorooctyl)oxy)buta (((3-(diethylamino)propoxy)carbonyl)oxy)-2-((((3-(diethylamino)propoxy)carbonyl)oxy) methyl)propyl (9Z,12Z)-octadeca-9,12-dienoate: 0.1 Intermediate IVf and 3-(diethylamino)propan-1-ol on a 1 mmol scale Prepared from. Yield 0.084g (71%). 1 H NMR (500 MHz, chloroform-d) δ 5. 43 - 5.28 (m, 4H), 4.48 (t, J = 5.5 Hz, 1H), 4.22 - 4.10 (m, 8H), 3.57 (dt, J = 9.3, 6.5 Hz, 2H), 3.41 (dt, J = 9.3, 6.5 Hz, 2H), 2.80 - 2.74 (m, 2H), 2.51 (q, J = 7.2 Hz, 6H), 2.45 - 2.36 (m, 3H), 2.31 (dd, J = 8.0, 7.2 Hz, 2H), 2.09 - 1.8 8 (m, 10H), 1.86 - 1.76 (m, 2H), 1.73 - 1.49 (m, 12H), 1.46 - 1.23 (m, 20H), 1.0 1 (t, J = 7.1 Hz, 6H), 0.92 - 0.86 (m, 3H). LCMS (Method A): (M+H) found m / z = 103 2.5, RT = 3.69 minutes.

[0316] [ka]

[0317] [Example 38] 3-((4,4-bis((7,7,8,8,8-pentafluorooctyl)oxy)buta ((((1-ethylpiperidin-3-yl)methoxy)carbo (9Z,12Z)-octadeca-9,12-dienoic acid Intermediate IVf and (1-ethylpiperidin-3-yl) ) Prepared from methanol. Yield 0.081 g (68%). 1 H NMR (500 MHz, chloroform) Lum-d) δ 5.43 - 5.28 (m, 4H), 4.49 (t, J = 5.5 Hz, 1H), 4.19 (d, J = 6.0 Hz, 2 H), 4.17 - 4.12 (m, 4H), 4.05 (dd, J = 10.6, 5.8 Hz, 1H), 3.96 (dd, J = 10.7, 7. 2 Hz, 1H), 3.57 (dt, J = 9.3, 6.6 Hz, 2H), 3.41 (dt, J = 9.3, 6.5 Hz, 2H), 2.90 (dd, J = 35.1, 11.3 Hz, 2H), 2.80 - 2.74 (m, 2H), 2.47 - 2.36 (m, 5H), 2.31 (dd, J = 8.0, 7.2 Hz, 2H), 2.11 - 1.79 (m, 11H), 1.79 - 1.66 (m, 3H), 1.66 - 1.49 (m , 12H), 1.49 - 1.23 (m, 24H), 1.08 (t, J = 7.2 Hz, 3H), 0.94 - 0.86 (m, 2H). LCM S (Method A): (M+H) found m / z = 1044.6, RT = 3.67 min.

[0318] [ka]

[0319] [Example 39] 3-((4,4-bis((7,7,8,8,8-pentafluorooctyl)oxy)buta 2-(((((1-methylpiperidin-4-yl)methoxy)carbonyl)-2-(((((1-methylpiperidin-4-yl)methoxy)carbonyl) (9Z,12Z)-octadeca-9,12-dienoic acid Intermediate IVf and (1-methylpiperidin-4-yl) ) Prepared from methanol. Yield 0.055 g (63%). LCMS (Method A): Observed (M+H) Value m / z = 1030.5, RT = 3.68 min.

[0320] [ka]

[0321] [Example 40] 3-((4,4-bis((7,7,8,8,8-pentafluorooctyl)oxy)buta (((3-(piperidin-1-yl)propoxy)carbonyl )Oxy)methyl)propyl (9Z,12Z)-octadeca-9,12-dienoate: Intermediate IVf and 3-(piperidin-1-yl)propane on a 0.09 mmol scale Prepared from -1-ol. Yield 0.052 g (58%). LCMS (Method A): Found (M+H) Value m / z = 1044.5, RT = 3.70 min.

[0322] [ka]

[0323] [Example 41] 3-((4,4-bis((7,7,8,8,8-pentafluorooctyl)oxy)buta 2-(((((octahydro-2H-quinolizin-1-yl)methoxy)-2-(((((octahydro-2H-quinolizin-1-yl)methoxy) (9Z,12Z)-octadeca-9,12-carbonyloxymethylpropyl Dienoates: Intermediate IVf and (octahydro-2H- Prepared from (quinolizin-1-yl)methanol. Yield 0.050g (55%). LCMS ( Method A): (M+H) found m / z = 1070.6, RT = 3.65 min.

[0324] [ka]

[0325] [Example 42] 3-((4,4-bis((7,7,8,8,8-pentafluorooctyl)oxy)buta (((3-(4-methylpiperazin-1-yl)propoxy)-2-((((3-(4-methylpiperazin-1-yl)propoxy) Carbonyl)oxy)methyl)propyl(9Z,12Z)-octadeca-9,12-dienes Noate: Intermediate IVf and 3-(4-methylpiperazine) on a 0.09 mmol scale Prepared from (-1-yl)propan-1-ol. Yield 0.049g (54%). LCMS ( Method A): (M+H) found m / z = 1059.5, RT = 3.66 min.

[0326] [ka]

[0327] [Example 43] 3-((4,4-bis((7,7,8,8,8-pentafluorooctyl)oxy)buta (((4-(diethylamino)butoxy)carbonyl)oxy)-2-((((4-(diethylamino)butoxy)carbonyl)oxy )Methyl)propyl (9Z,12Z)-octadeca-9,12-dienoate: 0.09 from intermediate IVf and 4-(diethylamino)butan-1-ol on a mmol scale Prepared. Yield 0.049 g (55%). LCMS (Method A): Found m / z for (M+H) = 1046.5, R T = 3.62 minutes.

[0328] [ka]

[0329] [Example 44] 3-((4,4-bis((7,7,8,8,8-pentafluorooctyl)oxy)buta (3-(3-(piperidin-1-yl)propoxy)prop (9Z,12Z)-octadeca-9,1 2-Dienoate: Intermediate IVf and 3-(3-(piperidine)-2-yl)-2-(dienoate) on a 0.09 mmol scale It was prepared from (di-(1-yl)propoxy)propan-1-ol. Yield: 0.044 g ( 46%). LCMS (Method A): (M+H) found m / z = 1102.5, RT = 3.66 min.

[0330] [ka]

[0331] [Example 45] 3-((4,4-bis((7,7,8,8,8-pentafluorooctyl)oxy)buta 2-(((((1,3-dimethylpiperidin-3-yl)methoxy)-2-(((((1,3-dimethylpiperidin-3-yl)oxy) Carbonyl)oxy)methyl)propyl(9Z,12Z)-octadeca-9,12-dienes IVf and (1,3-dimethylpiperidinyl) Prepared from (benzo-3-yl)methanol. Yield 0.042 g (70%). LCMS (Method A): Found m / z for (M+H) = 1044.5, RT = 3.68 min.

[0332] [ka]

[0333] [Example 46] 3-((4,4-bis((7,7,8,8,8-pentafluorooctyl)oxy)buta 4-(pyrrolidin-1-yl)butanoyl)oxy)methyl (9Z,12Z)-octadeca-9,12-dienoate: 0.9 mmol General Procedure from Intermediate IVf and 4-(Pyrrolidin-1-yl)butanoic Acid Hydrochloride Prepared according to E. Yield 0.079 g (91%). 1 H NMR (500 MHz, chloroform-d) δ 5.43 - 5.28 (m, 4H), 4.51 - 4.46 (m, 1H), 4.22 - 4.10 (m, 6H), 3.57 (dtd, J = 9.3, 6.6, 1.3 Hz, 2H), 3.41 (dt, J = 9.3, 6.5 Hz, 2H), 2.80 - 2.74 (m, 2H), 2. 55 - 2.27 (m, 12H), 2.11 - 1.72 (m, 15H), 1.69 - 1.52 (m, 16H), 1.47 - 1.23 (m, 18H), 0.92 - 0.86 (m, 3H). LCMS (Method A): (M+H) found m / z = 1014.4, RT = 3.68 min.

[0334] The following Examples 47 and 48 were prepared from intermediate IVg according to general procedure D.

[0335] [ka]

[0336] [Example 47] 3-((4,4-bis(4-(trifluoromethyl)phenethoxy)butanoyl)oxy) )-2-((((3-(diethylamino)propoxy)carbonyl)oxy)methyl)prop Dopyl(9Z,12Z)-octadeca-9,12-dienoate: 0.12 mmol scale It was prepared in vacuo from intermediate IVg and 3-(diethylamino)propan-1-ol. Yield 0.067 g (56%). LCMS (Method A): (M+H) found m / z = 972.5, RT = 3.58 min.

[0337] [ka]

[0338] [Example 48] 3-((4,4-bis(4-(trifluoromethyl)phenethoxy)butanoyl)oxy) )-2-(((((1-ethylpiperidin-3-yl)methoxy)carbonyl)oxy) Methyl)propyl (9Z,12Z)-octadeca-9,12-dienoate: 0.12m mol-scale from intermediate IVg and (1-ethylpiperidin-3-yl)methanol Prepared from. Yield 0.045 g (37%). LCMS (Method A): Found m / z for (M+H) = 984.5, RT = 3.56 minutes.

[0339] The following Examples 49 and 50 were prepared from intermediate IVh according to general procedure D.

[0340] [ka]

[0341] [Example 49] 3-((4,4-bis(3-cyclohexylpropoxy)butanoyl)oxy)-2-( (((3-(diethylamino)propoxy)carbonyl)oxy)methyl)propyl(9 Z,12Z)-Octadeca-9,12-dienoate: 0.14 mmol scale intermediate It was prepared from compound IVh and 3-(diethylamino)propan-1-ol. Yield: 0.0 59g (48%). LCMS (Method A): (M+H) found m / z = 876.6, RT = 4.08 min.

[0342] [ka]

[0343] [Example 50] 3-((4,4-bis(3-cyclohexylpropoxy)butanoyl)oxy)-2-( ((((1-ethylpiperidin-3-yl)methoxy)carbonyl)oxy)methyl)prop Dopyl(9Z,12Z)-octadeca-9,12-dienoate: 0.14 mmol was prepared in ethanol from intermediate IVh and (1-ethylpiperidin-3-yl)methanol. Yield 0.048 g (39%). LCMS (Method A): (M+H) found m / z = 888.7, RT = 4.04 Minutes.

[0344] The following Examples 51 and 52 were prepared according to general procedure E from intermediate IVh.

[0345] [ka]

[0346] [Example 51] 3-((4,4-bis(3-cyclohexylpropoxy)butanoyl)oxy)-2-( ((4-(pyrrolidin-1-yl)butanoyl)oxy)methyl)propyl(9Z,12 Z)-Octadeca-9,12-dienoate: intermediate IVh on a 0.14 mmol scale and 4-(pyrrolidin-1-yl)butanoic acid hydrochloride. Yield 0.104 g ( 87%). LCMS (Method A): (M+H) found m / z = 858.8, RT = 4.06 min.

[0347] [ka]

[0348] [Example 52] 3-((4,4-bis(3-cyclohexylpropoxy)butanoyl)oxy)-2-( ((3-(4-methylpiperazin-1-yl)propanoyl)oxy)methyl)propyl (9Z,12Z)-octadeca-9,12-dienoate: on a 0.14 mmol scale Prepared from intermediate IVh and 3-(4-methylpiperazin-1-yl)propanoic acid dihydrochloride Yield 0.074 g (61%). LCMS (Method A): (M+H) found m / z = 873.7, RT = 4.17 minutes.

[0349] [ka]

[0350] [Example 53] 3-((4,4-bis((3,7-dimethyloct-6-en-1-yl)oxy)buta 2-(((((1-ethylpyrrolidin-3-yl)methoxy)carbo (9Z,12Z)-octadeca-9,12-dienoic acid IVb and (1-ethylpyrrolidin-3-yl) ) Prepared from methanol according to general procedure D. Yield 40 mg (55%). LCMS (Method A) : (M+H) Found m / z = 902.7, RT = 3.99 min.

[0351] [ka]

[0352] [Example 54] 3-((4,4-bis((3,7-dimethyloct-6-en-1-yl)oxy)buta 1r,4r)-4-morpholinocyclohexyl)oxy)-2-((((((1r,4r)-4-morpholinocyclohexyl)oxy) Oxy)carbonyl)oxy)methyl)propyl(9Z,12Z)-octadeca-9,12 -dienoate: intermediate IVb and trans-4-mol on a 0.08 mmol scale Prepared from cyclohexyl methylpropional according to general procedure D. Yield 36 mg (4 7%). LCMS (Method A): (M+H) found m / z = 958.6, RT = 4.56 min.

[0353] [ka]

[0354] [Example 55] 3-((4,4-bis((3,7-dimethyloct-6-en-1-yl)oxy)buta (4-(4-methylpiperazin-1-yl)butoxy)-2-((((4-methylpiperazin-1-yl)butoxy) (9Z,12Z)-octadeca-9,12-dieno 4-(4-methylpiperazine-)-4-methylpiperazine ... Prepared from (1-yl)butan-1-ol according to general procedure D. Yield 27 mg (36%) LCMS (Method A): (M+H) found m / z = 945.7, RT = 3.95 min.

[0355] [ka]

[0356] [Example 56] 3-((4,4-bis((3,7-dimethyloct-6-en-1-yl)oxy)buta (((((1r,3s,5s)-8-methyl-8-azabicyclo[2.2.1.2.2.1]methyl)oxy)-2-( ...1r,3s,5s)-8-methyl-8-azabicyclo[2.2.1.2.1]methyl)oxy (ii)[3.2.1]octan-3-yl)oxy)carbonyl)oxy)methyl)propyl (9Z,12Z)-octadeca-9,12-dienoate: on a 0.08 mmol scale Prepared from intermediate IVb and pseudotropine according to general procedure D. Yield 19 mg ( 26%). LCMS (Method A): (M+H) found m / z = 914.8, RT = 3.88 min.

[0357] [ka]

[0358] [Example 57] 3-((4,4-bis((3,7-dimethyloct-6-en-1-yl)oxy)buta 4-morpholinobutoxy)carbonyl)oxy)methyl ) Propyl (9Z,12Z)-octadeca-9,12-dienoate: 0.08 mmol Follow general procedure D from intermediate IVb and 4-morpholinobutan-1-ol on a larger scale. Yield 39 mg (52%). LCMS (Method A): Found m / z for (M+H) = 932.7, RT = 4.58 minutes.

[0359] [ka]

[0360] [Example 58] 3-((4,4-bis((3,7-dimethyloct-6-en-1-yl)oxy)buta (2-(1-methylpiperidin-3-yl)ethoxy)-2-((((2-(1-methylpiperidin-3-yl)ethoxy) (9Z,12Z)-octadeca-9,12-dieno ester: intermediate IVb and 2-(1-methylpiperidine- Prepared from (3-yl)ethan-1-ol according to general procedure D. Yield 38 mg (52%) LCMS (Method A): (M+H) found m / z = 916.8, RT = 3.94 min.

[0361] [ka]

[0362] [Example 59] 3-((4,4-bis((3,7-dimethyloct-6-en-1-yl)oxy)buta 9Z,12Z)-octadeca-9,12-dienoyl)oxy)-2-((((9Z,12Z)-octadeca-9,12-dienoyl) Oxy)methyl)propyl 1'-ethyl-[1,4'-bipiperidine]-4-carboxy Rate: Intermediate IVb and 1'-ethyl-[1,4'-biphenyl] Prepared from [piperidine]-4-carboxylic acid dihydrochloride according to general procedure E. Yield 21 mg (27%). LCMS (Method A): (M+H) found m / z = 969.7, RT = 3.41 min.

[0363] [ka]

[0364] [Example 60] 3-((4,4-bis((3,7-dimethyloct-6-en-1-yl)oxy)buta 4-(dipropylamino)butanoyl)oxy)-2-(((4-(dipropylamino)butanoyl)oxy)methyl) Propyl (9Z,12Z)-octadeca-9,12-dienoate: 0.08 mmol From intermediate IVb and 4-(dipropylamino)butanoic acid hydrochloride to general procedure E Prepared as follows: Yield 62 mg (84%). LCMS (Method A): (M+H) found m / z = 916.8, RT = 4.00 minutes.

[0365] [ka]

[0366] [Example 61] 3-((4,4-bis((3,7-dimethyloct-6-en-1-yl)oxy)buta 9Z,12Z)-octadeca-9,12-dienoyl)oxy)-2-((((9Z,12Z)-octadeca-9,12-dienoyl) Oxy)methyl)propyl 1-(pyridin-4-yl)piperidine-4-carboxylate Intermediate IVb and 1-(pyridin-4-yl)piperidine were synthesized on a 0.08 mmol scale. Prepared from cinnam-4-carboxylic acid hydrochloride according to general procedure E. Yield 20 mg (27%) LCMS (Method A): (M+H) found m / z = 935.7, RT = 3.87 min.

[0367] [ka]

[0368] [Example 62] 3-((4,4-bis((3,7-dimethyloct-6-en-1-yl)oxy)buta 2-((2-(1-methylpiperidin-4-yl)acetoxy)methyl)- ... (9Z,12Z)-octadeca-9,12-dienoate: 0.08mmol 1-Methylpiperidin-4-yl)acetic acid from intermediate IVb and 2-(1-methylpiperidin-4-yl)acetic acid on a full scale Prepared according to procedure E. Yield 40 mg (56%). LCMS (Method A): Found m / z for (M+H) = 886.7, RT = 3.95 minutes.

[0369] [ka]

[0370] [Example 63] 3-((4,4-bis((3,7-dimethyloct-6-en-1-yl)oxy)buta 5-morpholinopentanoyl)oxy)-2-(((5-morpholinopentanoyl)oxy)methyl)propyl (9Z,12Z)-octadeca-9,12-dienoate: on a 0.08 mmol scale Prepared according to general procedure E from intermediate IVb and 5-morpholinopentanoic acid hydrochloride. Yield 64 mg (87%). LCMS (Method A): (M+H) found m / z = 916.9, RT = 4.50 min.

[0371] [ka]

[0372] [Example 64] 3-((4,4-bis((3,7-dimethyloct-6-en-1-yl)oxy)buta 5-(dimethylamino)pentanoyl)oxy)-2-(((5-(dimethylamino)pentanoyl)oxy)methyl) Propyl (9Z,12Z)-octadeca-9,12-dienoate: Example 63, 3-( (4,4-bis((3,7-dimethyloct-6-en-1-yl)oxy)butanoyl )oxy)-2-(((5-morpholinopentanoyl)oxy)methyl)propyl(9Z ,12Z)-Octadeca-9,12-dienoate: intermediate on a 0.08 mmol scale Prepared from IVb and 5-(dimethylamino)pentanoic acid hydrochloride according to general procedure E Yield 57 mg (81%). LCMS (Method A): (M+H) found m / z = 874.8, RT = 3.95 min.

[0373] [ka]

[0374] [Example 65] 3-((4,4-bis(((Z)-oct-5-en-1-yl)oxy)butanoyl) oxy)-2-((((2-(1-methylpiperidin-2-yl)ethoxy)carbonyl )Oxy)methyl)propyl (9Z,12Z)-octadeca-9,12-dienoate: Intermediate IVa and 2-(1-methylpiperidin-2-yl)- Prepared from (methyl)ethan-1-ol according to general procedure D. Yield 39 mg (52%). LC MS (Method A): (M+H) found m / z = 860.7, RT = 3.83 min.

[0375] [ka]

[0376] [Example 66] 3-(((3-(azetidin-1-yl)propoxy)carbonyl)oxy)-2-(( (4,4-bis(((Z)-oct-5-en-1-yl)oxy)butanoyl)oxy )Methyl)propyl (9Z,12Z)-octadeca-9,12-dienoate: 0.08 Intermediate IVa and 3-(azetidin-1-yl)propan-1-yl were synthesized on a 7 mmol scale. Prepared from ethanol according to general procedure D. Yield 41 mg (57%). LCMS (Method A): (M+H ) observed m / z = 832.6, RT = 3.74 min.

[0377] [ka]

[0378] [Example 67] 3-((4,4-bis((3,7-dimethyloct-6-en-1-yl)oxy)buta benzyl)oxy)-2-(((((2-((dimethylamino)methyl)benzyl)oxy )carbonyl)oxy)methyl)propyl(9Z,12Z)-octadeca-9,12-di Enoate: Intermediate IVb and (2-((dimethylamine)) on a 0.087 mmol scale Prepared from (methyl)phenyl)methanol according to general procedure D. Yield 30 mg (4 0%). LCMS (Method A): (M+H) found m / z = 938.7, RT = 4.28 min.

[0379] [ka]

[0380] [Example 68] 3-((4,4-bis((7,7,8,8,8-pentafluorooctyl)oxy)buta (((4-(diethylamino)butoxy)carbonyl)oxy)-2-((((4-(diethylamino)butoxy)carbonyl)oxy )Methyl)propyl (9Z,12Z)-octadeca-9,12-dienoate: 0.08 on a 6 mmol scale from intermediate IVf and 4-(diethylamino)butan-1-ol Prepared according to general procedure D from . Yield 49 mg (55%). LCMS (Method A): Observed (M+H) Value m / z = 1046.5, RT = 3.63 min.

[0381] [ka]

[0382] [Example 69] 3-((4,4-bis((7,7,8,8,8-pentafluorooctyl)oxy)buta (3-(3-(piperidin-1-yl)propoxy)prop (9Z,12Z)-octadeca-9,1 2-Dienoate: Intermediate IVf and 3-(3-(piperidine)-2-yl)-2-(dienoate) were prepared on a 0.086 mmol scale. Prepared from lysin-1-yl)propoxy)propan-1-ol according to general procedure D Yield 44 mg (46%). LCMS (Method A): (M+H) found m / z = 1102.5, RT = 3.66 min.

[0383] [ka]

[0384] [Example 70] 3-((4,4-bis(((Z)-oct-5-en-1-yl)oxy)butanoyl) oxy)-2-((((9Z,12Z)-octadeca-9,12-dienoyl)oxy) Methyl)propyl 1,3-dimethylpyrrolidine-3-carboxylate: 0.077mm The intermediate IVa and 1,3-dimethylpyrrolidine-3-carboxylic acid were converted to the basic Prepared according to procedure E. Yield 7 mg (11%). LCMS (Method A): Found m / z for (M+H) = 816.6, RT = 3.74 minutes.

[0385] [ka]

[0386] [Example 71] 3-((4,4-bis(((Z)-oct-5-en-1-yl)oxy)butanoyl) oxy)-2-((((3-(dimethylamino)propoxy)carbonyl)oxy)methyl (9Z,12Z)-octadeca-9,12-dienoate: 0.077mm The basic compound was prepared from intermediate IVa and 3-(dimethylamino)propan-1-ol on a 1 / 2 scale. Prepared according to procedure D. Yield 22 mg (35%). LCMS (Method A): Found m / z for (M+H) = 820.7, RT = 3.76 minutes.

[0387] [ka]

[0388] [Example 72] 3-((4,4-bis(((Z)-oct-5-en-1-yl)oxy)butanoyl) oxy)-2-((((3-(pyrrolidin-1-yl)propoxy)carbonyl)oxy )Methyl)propyl (9Z,12Z)-octadeca-9,12-dienoate: 0.07 Intermediate IVa and 3-(pyrrolidin-1-yl)propan-1-yl were synthesized on a 7 mmol scale. Prepared from ethanol according to general procedure D. Yield 27 mg (41%). LCMS (Method A): (M+H ) observed m / z = 846.7, RT = 3.73 min.

[0389] [ka]

[0390] [Example 73] 3-((4,4-bis((7,7,8,8,8-pentafluorooctyl)oxy)buta 2-(((((1,3-dimethylpiperidin-3-yl)methoxy)-2-(((((1,3-dimethylpiperidin-3-yl)oxy) Carbonyl)oxy)methyl)propyl(9Z,12Z)-octadeca-9,12-dienes IVf and (1,3-dimethylpiperidine) in a 0.057 mmol scale. Prepared from (di-(3-yl)-2-methyl-2-azabicyclo[4.2.1.2]methyl-1 ... LCMS (Method A): (M+H) found m / z = 1044.5, RT = 3.69 min.

[0391] [ka]

[0392] [Example 74] 2-(((4,4-bis(((Z)-oct-5-en-1-yl)oxy)butanoyl )oxy)methyl)-4-(((3-(diethylamino)propoxy)carbonyl)oxy butyl (9Z,12Z)-octadeca-9,12-dienoate

[0393] Step 1: 2-(hydroxymethyl)butane-1,4-diol

[0394] [ka]

[0395] Triethylethane-1,1,2-tricarboxylate (5 g, 20.3 mmol) A stirred solution of tert-butanol (80 mL) was added at 25 °C under an argon atmosphere. NaBH4 (2.3 g, 60.9 mmol) was added, and the resulting suspension was heated to reflux. Then, methanol (3 mL) was added dropwise in three portions within 30 minutes. The mixture was heated to reflux for 3 hours. The reaction mixture was then cooled to 25°C and diluted with 5N HCl (2. The precipitate was filtered and the filtrate was evaporated to give the crude material, which was by CombiFlash column chromatography eluting with 5% MeOH in DCM. Purification yielded 2-(hydroxymethyl)butane-1,4-diol (1.7 g, 69%) Obtained as a yellow liquid. 1 H NMR (400 MHz, DMSO-d6): δ1.34-1.46 (m, 2H), 1.49-1.63 (m, 1H), 3.27-3.48 (m, 6H), 4.34 (t, J = 5.2 Hz, 2H), 4.40 (t, J = 5.1 Hz, 1H).

[0396] Step 2: 2-(2,2-dimethyl-1,3-dioxan-5-yl)ethan-1-ol Rule

[0397] [ka]

[0398] 2-(hydroxymethyl)butane-1,4-diol (1.7 g, 14.1 mmol) and 2,2-dimethoxypropane (4.3 mL, 35.3 mmol) in THF (10 m A stirred solution of p-toluenesulfonic acid monohydrate was added to the solution at 25°C under an argon atmosphere. The reaction mixture was stirred at 25° C. for 16 hours. After this time, the reaction was neutralized with triethylamine (5 mL) and the solvent was removed under reduced pressure. The crude material was then purified by Combiflash column elution with 15% ethyl acetate-hexane. Purification by chromatography gave 2-(2,2-dimethyl-1,3-dioxane- 5-yl)ethan-1-ol (1.2 g, 53%) was obtained as a pale yellow liquid. 1 H NMR (400 MHz, CDCl3): δ1.41 (s, 6H), 1.50-1.60 (m, 2H), 1.88-1.98 (m, 1H), 3.63 (d d, J = 7.9, 11.8 Hz, 2H), 3.70 (t, J = 6.4 Hz, 2H), 3.93 (dd, J = 4.5, 11.8 Hz, 2H).

[0399] Step 3: 2-(2,2-dimethyl-1,3-dioxan-5-yl)ethyl(4- Nitrophenyl)carbonate

[0400] [ka]

[0401] 2-(2,2-dimethyl-1,3-dioxan-5-yl) ethoxylate in DCM (10 mL) A solution of tan-1-ol (400 mg, 2.5 mmol) was added to pyridine (0.4 mL, 5 0.0 mmol), DMAP (30.5 mg, 0.25 mmol), and finally chlorogyne 4-Nitrophenyl acetate (604 mg, 2.9 mmol) was added. The reaction was stirred at 25° C. for 7 After this time, the reaction mixture was diluted with water (20 mL) and DCM (30 mL). The organic layer was separated and the aqueous layer was extracted with DCM (10 mL x 2). The layer was dried over Na2SO4. The solvent was removed under reduced pressure to give a crude mass which was then diluted with 30% ethanol. Purification was performed by Combi-Flash column chromatography eluting with ethyl-hexane. , 2-(2,2-dimethyl-1,3-dioxan-5-yl)ethyl(4-nitrophenyl) The resulting product was tetrahydrofuran (TFA) carbonate (0.302 g, 37%) as a colorless liquid. 1 H NMR (400 MHz, C DCl3) δ 1.29-1.34 (m, 6H), 1.62 (q, J = 6.7 Hz, 2H), 1.80 (bs, 1H), 3.54 (q, J = 8.8 Hz, 2H), 3.83 (dd, J = 11.4 Hz, 4.0 Hz, 2H), 4.28 (t, J = 6.4 Hz, 2H), 7.5 7 (d, J = 9.0 Hz, 2H), 8.31 (d, J = 9.0 Hz, 2H).

[0402] Step 4: 3-(diethylamino)propyl (2-(2,2-dimethyl-1,3-diamino)propyl) Oxan-5-yl)ethyl)carbonate

[0403] [ka]

[0404] 2-(2,2-dimethyl-1,3-dioxan-5-yl)ethyl in 5mL DCM To a stirred solution of (4-nitrophenyl) carbonate (150 mg, 0.46 mmol), DMAP (5.6 mg, 0.04 mmol) and pyridine (0.07 mL, 0.9 mm ol) was added at 25°C and stirred for 5 minutes. Alcohol (78.7 mg, 0.6 mmol) was added at 25° C. The reaction mass was stirred at 25° C. for 9 hours. The reaction was judged complete by TLC (5% MeOH-DCM). The mixture was diluted with water and extracted with DCM (3 x 15 mL). The combined organic layers were washed with brine. The resulting mixture was washed, dried over anhydrous Na2SO4, filtered, and concentrated under reduced pressure. The crude material was purified by Combi-Flash column chromatography eluting with 10% MeOH-DCM. The product was purified by 3-Dioxan-5-yl)ethyl)carbonate (98 mg, 67%) as a pale yellow liquid And got it. 1 H NMR (400 MHz, CDCl3) δ 1.03 (t, J = 7.0 Hz, 3H), 1.40 (s, 3H), 1.6 3 (q, J = 6.7 Hz, 1H), 1.80-1.87 (m, 1H), 2.54 (t, J = 7.0 Hz, 3H), 3.58 (q, J = 8.2 Hz, 1H), 3.91 (dd, J = 11.8 Hz, 4.2 Hz, 1H), 4.13 (q, J = 6.2 Hz, 2H).

[0405] Step 5: 3-(diethylamino)propyl (4-hydroxy-3-(hydroxymethyl)amino)propyl (Thiyl)butyl)carbonate

[0406] [ka]

[0407] 3-(diethylamino)propyl (2-(2,2-dimethyl-) 1,3-Dioxan-5-yl)ethyl)carbonate (92 mg, 0.3 mmol) To the stirred solution, 1N HCl (0.9 mL, 0.9 mmol) was added at 25° C. The mixture was stirred for 2 hours and the reaction was judged complete by TLC (5% MeOH-DCM). The reaction mixture was concentrated and azeotroped twice with toluene to give the crude product, 3-(diethylamino)propyl 4-hydroxy-3-(hydroxymethyl)butyl carbonate (120 mg) It was used directly for the next step without purification.

[0408] Step 6: 4-(((3-(diethylamino)propoxy)carbonyl)oxy)- 2-(hydroxymethyl)butyl(9Z,12Z)-octadeca-9,12-dienoate to

[0409] [ka]

[0410] To a stirred solution of linoleic acid (0.08 mL, 0.26 mmol) in 3 mL DCM was added E DC (82.9 mg, 0.43 mmol) and DMAP (7.2 mg, 0.06 mmol) DIPEA (0.147 mL, 0.86 mmol) was added at 25°C and stirred for 5 min. l) and 3-(diethylamino)propyl (4-hydroxy-3-(hydroxymethyl )butyl)carbonate (80 mg, 0.29 mmol) was added at 25°C. The reaction mixture was stirred for 16 hours at 25° C. Completion of the reaction was confirmed by LCMS of the crude reaction mixture. The reaction mixture was diluted with water (10 mL) and extracted with DCM (15 mL × 3). The combined organic layers were washed with brine (10 mL), dried over anhydrous Na2SO4, and filtered. The crude material thus obtained was dissolved in 10% MeOH-DCM. The compound was purified by combi-flash column chromatography to give 4-(((3-( Diethylamino)propoxy)carbonyl)oxy)-2-(hydroxymethyl)butyl (9Z,12Z)-Octadeca-9,12-dienoate (42 mg, 27%) was obtained as a pale yellow Obtained as a liquid. 1 H NMR (400 MHz, CDCl3) δ 0.87 (d, J = 6.6 Hz, 3H), 1.00-1.10 ( m, 5H), 1.24-1.29 (m, 20H), 1.69-1.83 (m, 4H), 2.02 (t, J = 6.6 Hz, 5H), 2.30 (t , J = 7.3 Hz, 2H), 2.50-2.55 (m, 5H), 2.75 (d, J = 7.3 Hz, 2H), 3.54-3.61 (m, 2H ), 4.13-4.23 (m, 5H), 5.34 (t, J = 4.5 Hz, 4H).

[0411] Step 7: 2-(((4,4-bis(((Z)-oct-5-en-1-yl))oxo (3-(diethylamino)propoxy)methyl)-4-( ... (carbonyl)oxy)butyl (9Z,12Z)-octadeca-9,12-dienoate (actual Example 74: 4,4-bis(((Z)-oct-5-en-1-yl)methylpropional in 2 mL DCM To a stirred solution of 24.1 mg (0.07 mmol) of hydroxybutanoic acid (intermediate IIa) , EDC (22.3 mg, 0.12 mmol) and DMAP (1.9 mg, 0.015 0.04 mL of DIPEA (0.05 mmol) was added at 25° C. and stirred for 5 min. 0.23 mmol) and 4-(((3-(diethylamino)propoxy)carbonyl)o (9Z,12Z)-octadeca-9,12-dihydroxy-2-(hydroxymethyl)butyl The enoate (42 mg, 0.08 mmol) was added at 25° C. The reaction mixture was stirred at 25° C. Stirred for 16 hours. The reaction was judged complete by LCMS of the crude reaction mixture. The mixture was diluted with NaHCO3 solution (5 mL) and extracted with DCM (10 mL x 3). The organic layer was washed with brine (10 mL), dried over anhydrous Na2SO4, filtered, and evaporated under reduced pressure. The crude material thus obtained was purified by Prep-HPLC. , -(((4,4-bis(((Z)-oct-5-en-1-yl)oxy)butanoyl )oxy)methyl)-4-(((3-(diethylamino)propoxy)carbonyl)oxy butyl (9Z,12Z)-octadeca-9,12-dienoate (24 mg, 36%) ) was obtained as a pale yellow liquid. LCMS (Method B): (M+H) found m / z = 867.7, RT = 2.04 min.

[0412] [ka]

[0413] [Example 75] 3-((4,4-bis(((Z)-oct-5-en-1-yl)oxy)butanoyl) oxy)-2-(((3-(4-ethylpiperazin-1-yl)propanoyl)oxy) Methyl)propyl (9Z,12Z)-octadeca-9,12-dienoate

[0414] Step 1: 3-(acryloyloxy)-2-(((4,4-bis(((Z)-octyloxy)-4,4-di ... ter-5-en-1-yl)oxy)butanoyl)oxy)methyl)propyl(9Z,12 Z)-Octadeca-9,12-dienoate (Intermediate V)

[0415] [ka]

[0416] 3-((4,4-bis(((Z)-oct-5-en-1-yl)methyl)-2- ... 2-(hydroxymethyl)propyl (9Z, 12Z)- ... )-Octadeca-9,12-dienoate (Intermediate IVa) (1.0 g, 1.4 mmol ), DMAP (18.0 mg, 0.1 mmol) and DIPEA (0.5 The resulting mixture was cooled in an ice bath and then acetyl chloride was added. Trimethylsilyl (0.2 mL, 2.9 mmol) was added dropwise over 10 minutes. The mixture was allowed to stand at room temperature. The mixture was stirred for 1 hour. After this time, the reaction mixture was extracted with DCM (20 mL x 2). The combined organic layers were washed with water (20 mL × 2) and brine (20 mL × 2) and diluted with anhydrous N The crude material was dried over Na2SO4, filtered, and concentrated under reduced pressure. The compound was purified by CombiFlash column chromatography eluting with 2-(2-methyl-2-propanol)-2-one to give 3-((2-methyl-2-propanol)-2-one. ((4,4-bis(((Z)-oct-5-en-1-yl)oxy)-2-(((4,4-bis(((Z)-oct-5-en-1-yl) Oxy)butanoyl)oxy)methyl)propyl(9Z,12Z)-octadeca-9,1 The 2-dienoate (860 mg, 79%) was obtained as a colorless liquid. 1 H NMR (400 MHz, C12H-d) δ 0.84 - 0.91 (m, 3H), 0.94 (t, J = 7.5 Hz, 6H), 1.17 - 1.47 (m, 20H), 1.53 - 1.65 (m, 4H), 1.86 - 1.95 (m, 2H), 1.97 - 2.09 (m, 12H), 2.30 (t, J = 7.4 Hz, 2H), 2.34 - 2.47 (m, 3H), 2.76 (t, J = 6.4 Hz, 2H), 3.34 - 3.45 (m, 2 H), 3.56 (q, J = 7.4 Hz, 2H), 4.11 - 4.18 (m, 4H), 4.22 (d, J = 6.0 Hz, 2H), 4.4 7 (t, J = 5.6 Hz, 1H), 5.22 - 5.45 (m, 8H), 5.85 (d, J = 11.3 Hz, 1H), 6.11 (dd, J = 10.5, 17.3 Hz, 1H), 6.36-6.45 (m, 1H).

[0417] Step 2: 3-((4,4-bis(((Z)-oct-5-en-1-yl)oxy) )butanoyl)oxy)-2-(((3-(4-ethylpiperazin-1-yl)propano yl)oxy)methyl)propyl(9Z,12Z)-octadeca-9,12-dienoate Example 75: The following is representative of general procedure F. 4,4-bis(((Z)-oct-5-en-1-yl)oxy)buta (9Z,12Z)-octadeca-9,12-dienoyl)oxy)methyl)propyl Intermediate V (50 mg, 0.067 mmol, 1 Eq) and 1-ethylpiperazine A mixture of 1,2-dimethyl-2,4-trimethyl-1,4-trimethyl ... After cooling to room temperature, the crude reaction mixture was eluted with 9:1 DCM / MeOH. Purification by column chromatography gave 3-((4,4-bis(((Z)-octadecanoate). -5-en-1-yl)oxy)butanoyl)oxy)-2-(((3-(4-ethylpyridinyl)oxy) Perazin-1-yl)propanoyl)oxy)methyl)propyl(9Z,12Z)-octyl Tadeca-9,12-dienoate (26 mg, 44%) was obtained. LCMS (Method B): (M+H) Found m / z = 859.9, RT = 1.67 min.

[0418] [ka]

[0419] [Example 76] 3-((4,4-bis(((Z)-oct-5-en-1-yl)oxy)butanoyl) oxy)-2-(((3-(4-ethyl-1,4-diazepan-1-yl)propanoyl )Oxy)methyl)propyl (9Z,12Z)-octadeca-9,12-dienoate: Prepared from intermediate V and 1-ethyl-1,4-diazepane according to general procedure F. Yield 16 mg (46%). LCMS (Method B): (M+H) found m / z = 873.9, RT = 1.56 min.

[0420] [ka]

[0421] [Example 77] 3-((4,4-bis(((Z)-oct-5-en-1-yl)oxy)butanoyl) oxy)-2-(((3-(4-(2-methoxyethyl)piperazin-1-yl)propanol (9Z,12Z)-octadeca-9,12-dienoyl)oxy)methyl)propyl Prepared from intermediate V and 1-(2-methoxyethyl)piperazine according to general procedure F. Yield: 34 mg (57%). LCMS (Method B): (M+H) found m / z = 889.9, RT = 1.6 9 minutes.

[0422] [ka]

[0423] [Example 78] 3-((4,4-bis(((Z)-oct-5-en-1-yl)oxy)butanoyl) oxy)-2-(((3-(3,5-dimethylpiperazin-1-yl)propanoyl)o (Oxy)methyl)propyl (9Z,12Z)-octadeca-9,12-dienoate: intermediate Prepared from compound V and 2,6-dimethylpiperazine according to general procedure F. Yield: 38 mg (65%). LCMS (Method B): (M+H) found m / z = 859.9, RT = 1.71 min.

[0424] [ka]

[0425] [Example 79] 3-((4,4-bis(((Z)-oct-5-en-1-yl)oxy)butanoyl) oxy)-2-(((3-((2-(diethylamino)ethyl)(ethyl)amino)propanol (9Z,12Z)-octadeca-9,12-dieno Etate: from intermediate V and N1,N1,N2-triethylethane-1,2-diamine Prepared according to procedure F. Yield 24 mg (48%). LCMS (Method B): Found m / z for (M+H) = 889.9, RT = 2.29 minutes.

[0426] [ka]

[0427] [Example 80] 3-((4,4-bis(((Z)-oct-5-en-1-yl)oxy)butanoyl) oxy)-2-(((3-((1-isopropylpiperidin-4-yl)amino)propanol (9Z,12Z)-octadeca-9,12-dienoyl)oxy)methyl)propyl Preparation of intermediate V from 1-isopropylpiperidin-4-amine according to general procedure F Prepared. Yield 18 mg (37%). LCMS (Method B): (M+H) found m / z = 887.9, RT = 1 .34 minutes.

[0428] [ka]

[0429] [Example 81] 3-((4,4-bis(((Z)-oct-5-en-1-yl)oxy)butanoyl) oxy)-2-(((3-((pyridin-3-ylmethyl)amino)propanoyl)oxy (Ci)methyl)propyl (9Z,12Z)-octadeca-9,12-dienoate: intermediate Prepared from V and pyridin-3-ylmethanamine according to general procedure F. Yield 20 ml g (43%). LCMS (Method B): (M+H) found m / z = 854.0, RT = 1.34 min.

[0430] [ka]

[0431] [Example 82] 3-((4,4-bis(((Z)-oct-5-en-1-yl)oxy)butanoyl) oxy)-2-(((3-(methyl(2-(pyridin-2-yl)ethyl)amino)propanol (9Z,12Z)-octadeca-9,12-dieno Ethyl: from intermediate V and N-methyl-2-(pyridin-2-yl)ethan-1-amine Prepared according to general procedure F from . Yield 34 mg (49%). LCMS (Method B): Observed (M+H) Value m / z = 882.0, RT = 1.07 min.

[0432] [ka]

[0433] [Example 83] 3-((4,4-bis(((Z)-oct-5-en-1-yl)oxy)butanoyl) oxy)-2-(((3-(3-(dimethylamino)pyrrolidin-1-yl)propanoyl (9Z,12Z)-octadeca-9,12-dienoate : Prepared from intermediate V and 3-(dimethylamino)pyrrolidine according to general procedure F. Yield 27 mg (58%). LCMS (Method B): (M+H) found m / z = 859.9, RT = 1.62 min.

[0434] [ka]

[0435] [Example 84] 3-((4,4-bis(((Z)-oct-5-en-1-yl)oxy)butanoyl) oxy)-2-(((3-((2-(pyrrolidin-1-yl)ethyl)amino)propano yl)oxy)methyl)propyl(9Z,12Z)-octadeca-9,12-dienoate General Procedure F: From Intermediate V and 2-(pyrrolidin-1-yl)ethan-1-amine Prepared as follows: Yield 26 mg (56%). LCMS (Method B): (M+H) found m / z = 859.9, RT = 1.34 minutes.

[0436] [ka]

[0437] [Example 85] 3-((4,4-bis(((Z)-oct-5-en-1-yl)oxy)butanoyl) oxy)-2-(((3-((3-morpholinopropyl)amino)propanoyl)oxy )Methyl)propyl (9Z,12Z)-octadeca-9,12-dienoate: Intermediate V and 2-(1-methylpyrrolidin-2-yl)ethan-1-amine according to general procedure F. Yield 17 mg (35%). LCMS (Method B): (M+H) found m / z = 873.8, RT = 2.28 minutes.

[0438] [ka]

[0439] [Example 86] 3-((4,4-bis(((Z)-oct-5-en-1-yl)oxy)butanoyl) oxy)-2-(((3-((2-(pyridin-4-yl)ethyl)amino)propanoyl (9Z,12Z)-octadeca-9,12-dienoate : From intermediate V and 2-(pyridin-4-yl)ethan-1-amine according to general procedure F Yield 18 mg (39%). LCMS (Method B): (M+H) found m / z = 867.9, RT = 1.34 minutes.

[0440] [ka]

[0441] [Example 87] 3-((4,4-bis(((Z)-oct-5-en-1-yl)oxy)butanoyl) oxy)-2-(((3-((2-(1-methylpyrrolidin-2-yl)ethyl)amino )propanoyl)oxy)methyl)propyl(9Z,12Z)-octadeca-9,12- Dienoates: From intermediate V and 2-(pyridin-4-yl)ethan-1-amine Prepared according to procedure F. Yield 26 mg (43%). LCMS (Method B): Found m / z for (M+H) = 867.9, RT = 1.34 minutes.

[0442] [ka]

[0443] [Example 88] 3-((4,4-bis(((Z)-non-3-en-1-yl)oxy)butanoyl)o oxy)-2-((((3-(diethylamino)propoxy)carbonyl)oxy)methyl ) Propyl (9Z,12Z)-octadeca-9,12-dienoate

[0444] Step 1: 4,4-bis(((Z)-non-3-en-1-yl)oxy)butanediol Trill

[0445] [ka]

[0446] Prepared according to general procedure A using (Z)-non-3-en-1-ol. Yield 2 80mg (41%). 1 H NMR (400 MHz, chloroform-d): δ 0.88 (t, J = 6.7 Hz, 6H ), 1.21-1.41 (m, 12H), 1.94 (q, J = 7.2 Hz, 2H), 2.03 (q, J = 7.2 Hz, 4H), 2.32 (q, J = 7.0 Hz, 4H), 2.41 (t, J = 7.4 Hz, 2H), 3.39-3.50 (m, 2H), 3.56-3.66 (m, 2H), 4.58 (t, J = 5.3 Hz, 1H), 5.29-5.40 (m, 2H), 5.41-5.53 (m, 2H).

[0447] Step 2: 4,4-bis(((Z)-non-3-en-1-yl)oxy)butanoic acid

[0448] [ka]

[0449] 4,4-bis(((Z)-non-3-en-1-yl)oxy)butanenitrile as a base Prepared according to procedure B. Yield 230 mg (78%). 1 H NMR (400 MHz, DMSO-d6): δ 0.85 (t, J = 6.6 Hz, 6H), 1.13-1.38 (m, 13H), 1.72 (q, J = 7.1 Hz, 2H), 2.00 (q, J = 7.0 Hz, 4H), 2.16-2.29 (m, 5H), 3.33-3.42 (m, 2H), 3.43-3.54 (m, 2H), 4. 49 (t, J = 5.6 Hz, 1H), 5.30-5.47 (m, 4H), 12.06 (s, 1H).

[0450] Step 3: 3-((4,4-bis(((Z)-non-3-en-1-yl)oxy) Butanoyl)oxy)-2-(hydroxymethyl)propyl(9Z,12Z)-octadeca Carboxy-9,12-dienoate

[0451] [ka]

[0452] General Procedure from 4,4-bis(((Z)-non-3-en-1-yl)oxy)butanoic Acid Prepared according to C. Yield 340 mg (58%). 1 H NMR (400 MHz, chloroform-d): δ 0.82-0.92 (m, 9H), 1.21-1.40 (m, 26H), 1.61 (t, J = 7.1 Hz, 2H), 1.88 - 2.09 (m, 10H), 2.13 - 2.23 (m, 2H), 2.30 (q, J = 7.0 Hz, 6H), 2.41 (t, J = 7.5 Hz, 2H ), 2.76 (t, J = 6.4 Hz, 2H), 3.36 - 3.47 (m, 2H), 3.52 - 3.65 (m, 4H), 4.13 - 4. 23 (m, 4H), 4.52 (t, J = 5.5 Hz, 1H), 5.26 - 5.51 (m, 8H).

[0453] Step 4: 3-((4,4-bis(((Z)-non-3-en-1-yl)oxy) butanoyl)oxy)-2-((((3-(diethylamino)propoxy)carbonyl) Oxy)methyl)propyl (9Z,12Z)-octadeca-9,12-dienoate (actual Example 88) 3-((4,4-bis(((Z)-non-3-en-1-yl)oxy)butanol (9Z,12Z)-octadeca(2-hydroxymethyl)propyl 2-( ... -9,12-dienoate and 3-(diethylamino)propan-1-ol to form the basic Prepared according to procedure D. Yield 79 mg (83%). LCMS (Method B): Found m / z for (M+H) = 876.9, RT = 1.91 minutes.

[0454] [ka]

[0455] [Example 89] 3-((4,4-bis(((Z)-non-3-en-1-yl)oxy)butanoyl)o oxy)-2-(((((1-ethylpiperidin-3-yl)methoxy)carbonyl)oxy 3-(methyl)propyl (9Z,12Z)-octadeca-9,12-dienoate (4,4-bis(((Z)-non-3-en-1-yl)oxy)butanoyl)oxy) -2-(hydroxymethyl)propyl(9Z,12Z)-octadeca-9,12-dieno Prepared from 1-ethylpiperidin-3-yl)methanol according to general procedure D. Yield: 50 mg (69%). LCMS (Method B): (M+H) found m / z = 889.0, RT = 1.7 7 minutes.

[0456] [ka]

[0457] [Example 90] 3-((4,4-bis((4-butylbenzyl)oxy)butanoyl)oxy)-2-( (((3-(diethylamino)propoxy)carbonyl)oxy)methyl)propyl(9 Z,12Z)-Octadeca-9,12-dienoate

[0458] Step 1: 4,4-bis((4-butylbenzyl)oxy)butanenitrile

[0459] [ka]

[0460] Prepared according to general procedure A using (4-butylphenyl)methanol. Yield 1. 1g (36%). 1 H NMR (400 MHz, chloroform-d): δ 0.92 (t, J = 7.3 Hz, 6H), 1 .28 - 1.40 (m, 4H), 1.51 - 1.65 (m, 4H), 1.97 - 2.07 (m, 2H), 2.41 (t, J = 7.4 H z, 2H), 2.60 (t, J = 7.7 Hz, 4H), 4.51 (d, J = 11.5 Hz, 2H), 4.64 (d, J = 11.5 H z, 2H), 4.79 (t, J = 5.3 Hz, 1H), 7.16 (d, J = 7.9 Hz, 4H), 7.20 - 7.28 (m, 4H).

[0461] Step 2: 4,4-bis((4-butylbenzyl)oxy)butanoic acid

[0462] [ka]

[0463] From 4,4-bis((4-butylbenzyl)oxy)butanenitrile according to general procedure B The yield was 1.1 g (91%). 1 H NMR (400 MHz, DMSO-d6): δ 0.89 (t, J = 7 .3 Hz, 6H), 1.22 - 1.36 (m, 4H), 1.47 - 1.60 (m, 4H), 1.87 (q, J = 7.2 Hz, 2H), 2.27 (t, J = 7.4 Hz, 2H), 2.56 (t, J = 7.6 Hz, 4H), 4.45 (d, J = 11.7 Hz, 2H), 4 .56 (d, J = 11.7 Hz, 2H), 4.72 (t, J = 5.5 Hz, 1H), 7.15 (d, J = 7.9 Hz, 4H), 7. 22 (d, J = 7.6 Hz, 4H), 12.09 (s, 1H).

[0464] Step 3: 3-((4,4-bis((4-butylbenzyl)oxy)butanoyl)oxy) (9Z,12Z)-octadeca-9,12-oxy)-2-(hydroxymethyl)propyl Dienoate

[0465] [ka]

[0466] Prepared from 4,4-bis((4-butylbenzyl)oxy)butanoic acid according to general procedure C The yield was 400 mg (64%). 1 H NMR (400 MHz, chloroform-d): δ 0.83 - 0.96 (m, 9H), 1.21 - 1.42 (m, 17H), 1.52 - 1.66 (m, 7H), 1.99 - 2.19 (m, 8H), 2.30 ( t, J = 7.6 Hz, 2H), 2.44 (t, J = 7.4 Hz, 2H), 2.59 (t, J = 7.7 Hz, 4H), 2.76 (t, J = 6.4 Hz, 2H), 3.55 (t, J = 5.7 Hz, 2H), 4.04 - 4.19 (m, 4H), 4.50 (d, J = 11 .5 Hz, 2H), 4.61 (d, J = 11.5 Hz, 2H), 4.74 (t, J = 5.4 Hz, 1H), 5.26 - 5.42 (m, 4H), 7.15 (d, J = 7.9 Hz, 4H), 7.22 (d, J = 7.8 Hz, 4H).

[0467] Step 4: 3-((4,4-bis((4-butylbenzyl)oxy)butanoyl)oxy) oxy)-2-((((3-(diethylamino)propoxy)carbonyl)oxy)methyl ) Propyl (9Z,12Z)-octadeca-9,12-dienoate (Example 90). 3 -((4,4-bis((4-butylbenzyl)oxy)butanoyl)oxy)-2-(hydroxybenzoyl) (hydroxymethyl)propyl (9Z,12Z)-octadeca-9,12-dienoate and and 3-(diethylamino)propan-1-ol according to general procedure D. 62 mg (83%). LCMS (Method B): (M+H) found m / z = 920.9, RT = 1.81 min.

[0468] [ka]

[0469] [Example 91] 3-((4,4-bis((4-butylbenzyl)oxy)butanoyl)oxy)-2-( ((((1-ethylpiperidin-3-yl)methoxy)carbonyl)oxy)methyl)prop Lopyl(9Z,12Z)-octadeca-9,12-dienoate. 3-((4,4-bis((4-butylbenzyl)oxy)butanoyl)oxy)-2- (Hydroxymethyl)propyl (9Z,12Z)-octadeca-9,12-dienoate and (1-ethylpiperidin-3-yl)methanol according to general procedure D Yield 50 mg, 69%. LCMS (Method B): (M+H) found m / z = 889.0, RT = 1.77 min.

[0470] [ka]

[0471] [Example 92] 3-((4,4-bis(((Z)-dec-4-en-1-yl)oxy)butanoyl)o oxy)-2-((((3-(diethylamino)propoxy)carbonyl)oxy)methyl ) Propyl (9Z,12Z)-octadeca-9,12-dienoate

[0472] Step 1: 4,4-bis(((Z)-dec-4-en-1-yl)oxy)butanediol Trill

[0473] [ka]

[0474] Prepared according to general procedure A using (Z)-dec-4-en-1-ol. Yield 7 50mg (51%). 1H NMR (400 MHz, DMSO-d6): δ 0.86 (t, J = 6.8 Hz, 6H), 1.18 - 1.37 (m, 12H), 1.48 - 1.60 (m, 4H), 1.74 - 1.86 (m, 2H), 1.94 - 2.10 (m, 8H), 2.46 (t, J = 7.2 Hz, 2H), 3.33 - 3.44 (m, 2H), 3.47 - 3.60 (m, 2H), 4.51 (t, J = 5.4 Hz, 1H), 5.28 - 5.42 (m, 4H).

[0475] Step 2: 4,4-bis(((Z)-dec-4-en-1-yl)oxy)butanoic acid

[0476] [ka]

[0477] 4,4-bis(((Z)-dec-4-en-1-yl)oxy)butanenitrile as a base Prepared according to procedure B. Yield 300 mg (71%). 1 H NMR (400 MHz, DMSO-d6): δ 0.85 (t, J = 6.7 Hz, 6H), 1.19 - 1.36 (m, 12H), 1.53 (q, J = 6.9 Hz, 4H), 1.7 3 (q, J = 7.0 Hz, 2H), 1.91 - 2.09 (m, 8H), 2.22 (t, J = 7.5 Hz, 2H), 3.29 - 3.4 0 (m, 2H), 3.44 - 3.55 (m, 2H), 4.45 (t, J = 5.5 Hz, 1H), 5.27 - 5.42 (m, 4H), 1 2.05 (s, 1H).

[0478] Step 3: 3-((4,4-bis(((Z)-dec-4-en-1-yl)oxy) Butanoyl)oxy)-2-(hydroxymethyl)propyl(9Z,12Z)-octadeca Carboxy-9,12-dienoate

[0479] [ka]

[0480] General Procedure from 4,4-bis(((Z)-dec-4-en-1-yl)oxy)butanoic Acid Prepared according to C. Yield 270 mg (47%). 1 H NMR (400 MHz, chloroform-d): δ 0.83 - 0.92 (m, 9H), 1.07 - 1.15 (m, 2H), 1.20 - 1.41 (m, 23H), 1.47 - 1.76 ( m, 9H), 1.88 - 2.13 (m, 10H), 2.17 - 2.23 (m, 2H), 2.31 (t, J = 7.5 Hz, 2H), 2.4 1 (t, J = 7.4 Hz, 2H), 2.76 (t, J = 6.6 Hz, 2H), 3.35 - 3.50 (m, 3H), 3.52 - 3.6 5 (m, 4H), 3.97 - 4.06 (m, 1H), 4.10 - 4.22 (m, 4H), 4.48 (t, J = 5.4 Hz, 1H), 5 .26 - 5.44 (m, 8H).

[0481] Step 4: 3-((4,4-bis(((Z)-dec-4-en-1-yl)oxy) butanoyl)oxy)-2-((((3-(diethylamino)propoxy)carbonyl) Oxy)methyl)propyl (9Z,12Z)-octadeca-9,12-dienoate (actual Example 92) 3-((4,4-bis(((Z)-dec-4-en-1-yl)oxy)butanol (9Z,12Z)-octadeca(2-hydroxymethyl)propyl 2-( ... The basic procedure is as follows: Prepared according to procedure D. Yield 40 mg (61%). LCMS (Method B): Found m / z for (M+H) = 9 05.0, RT = 1.90 minutes.

[0482] [ka]

[0483] [Example 93] 3-((4,4-bis(((Z)-dec-4-en-1-yl)oxy)butanoyl)o oxy)-2-(((((1-ethylpiperidin-3-yl)methoxy)carbonyl)oxy 3-(methyl)propyl (9Z,12Z)-octadeca-9,12-dienoate (4,4-bis(((Z)-dec-4-en-1-yl)oxy)butanoyl)oxy) -2-(hydroxymethyl)propyl(9Z,12Z)-octadeca-9,12-dieno Prepared from 1-ethylpiperidin-3-yl)methanol according to general procedure D. Yield: 50 mg (68%). LCMS (Method B): (M+H) found m / z = 917.0, RT = 1.9 1 minute.

[0484] [ka]

[0485] [Example 94] 3-(((3-(diethylamino)propoxy)carbonyl)oxy)-2-(((3- (5,5-Dioctyl-1,3-dioxan-2-yl)propanoyl)oxy)methyl ) Propyl (9Z,12Z)-octadeca-9,12-dienoate

[0486] Step 1: 3-(5,5-dioctyl-1,3-dioxan-2-yl)propanediol Trill

[0487] [ka]

[0488] Prepared according to general procedure A using 2,2-dioctylpropane-1,3-diol Yield: 500 mg (70%). 1 H NMR (400 MHz, chloroform-d): δ 0.83-0.91 (m, 6H), 0.92-1.02 (m, 2H), 1.05-1.35 (m, 24H), 1.53-1.63 (m, 2H), 1.89-1.99 (m, 2H) ), 2.47 (t, J = 7.4 Hz, 2H), 3.37 (d, J = 11.1 Hz, 2H), 3.76 (d, J = 11.1 Hz, 2H ), 4.55 (t, J = 4.6 Hz, 1H).

[0489] Step 2: 3-(5,5-dioctyl-1,3-dioxan-2-yl)propanoic acid

[0490] [ka]

[0491] 3-(5,5-dioctyl-1,3-dioxan-2-yl)propanenitrile to form the base Prepared according to procedure B. Yield 400 mg (70%). 1H NMR (400 MHz, DMSO-d6): δ 0.78-0.97 (m, 8H), 1.02-1.35 (m, 24H), 1.47-1.57 (m, 2H), 1.67-1.76 (m, 2H), 2.26 (t, J = 7.5 Hz, 2H), 3.33 (d, J = 10.3 Hz, 2H), 3.67 (d, J = 10.9 Hz, 2H), 4.44 (t, J = 4.8 Hz, 1H), 12.05 (s, 1H).

[0492] Step 3: 3-((3-(5-hexyl-5-octyl-1,3-dioxane-2- 2-(hydroxymethyl)propyl(9Z,12Z)- Octadeca-9,12-dienoate

[0493] [ka]

[0494] General Procedure from 3-(5,5-Dioctyl-1,3-dioxan-2-yl)propanoic Acid Prepared according to C. Yield 80 mg (40%). 1 H NMR (400 MHz, chloroform-d): δ 0.79-1.01 (m, 11H), 1.02-1.45 (m, 38H), 1.56-1.65 (m, 4H), 1.89-1.99 (m, 2H), 1 .99-2.09 (m, 4H), 2.12-2.26 (m, 2H), 2.31 (t, J = 7.6 Hz, 2H), 2.47 (t, J = 7.5 Hz, 2H), 2.76 (t, J = 6.3 Hz, 2H), 3.34 (d, J = 11.0 Hz, 2H), 3.60 (t, J = 6.0 H z, 2H), 3.74 (d, J = 11.1 Hz, 2H), 4.17 (t, J = 7.1 Hz, 4H), 4.42-4.54 (m, 1H), 5:30-5:39 (m, 4H).

[0495] Step 4: 3-(((3-(diethylamino)propoxy)carbonyl)oxy)- 2-(((3-(5,5-dioctyl-1,3-dioxan-2-yl)propanoyl) Oxy)methyl)propyl (9Z,12Z)-octadeca-9,12-dienoate (actual Example 94) 3-((3-(5-hexyl-5-octyl-1,3-dioxane-2-yl) (9Z,12Z)-2-(hydroxymethyl)propyl (9Z,12Z)- From ctadeca-9,12-dienoate and 3-(diethylamino)-1-propanol Prepared according to general procedure D from . Yield 27 mg (73%). LCMS (Method B): Observed (M+H) Value m / z = 892.9, RT = 1.93 min.

[0496] [ka]

[0497] [Example 95] 3-((3-(5,5-dioctyl-1,3-dioxan-2-yl)propanoyl)o oxy)-2-(((((1-ethylpiperidin-3-yl)methoxy)carbonyl)oxy 3-(methyl)propyl (9Z,12Z)-octadeca-9,12-dienoate (3-(5-hexyl-5-octyl-1,3-dioxan-2-yl)propanoyl) Oxy)-2-(hydroxymethyl)propyl(9Z,12Z)-octadeca-9,12 -dienoate and (1-ethylpiperidin-3-yl)methanol to general procedure D Prepared as follows: Yield 27 mg (59%). LCMS (Method B): (M+H) found m / z = 905.0. RT = 1.94 minutes.

[0498] [ka]

[0499] [Example 96] 3-((4,4-bis(2-(octyloxy)ethoxy)butanoyl)oxy)-2- ((((3-(diethylamino)propoxy)carbonyl)oxy)methyl)propyl( 9Z,12Z)-Octadeca-9,12-dienoate

[0500] Step 1: 4,4-bis(2-(octyloxy)ethoxy)butanenitrile

[0501] [ka]

[0502] Prepared according to general procedure A using 2-(octyloxy)ethan-1-ol. Yield: 340 mg (21%). 1 H NMR (400 MHz, chloroform-d): δ 0.87 (t, J = 6.7 Hz, 6H), 1.23 - 1.37 (m, 20H), 1.53 (d, J = 7.7 Hz, 2H), 1.58 (d, J = 7.8 Hz, 2H) ), 1.93 - 2.03 (m, 2H), 2.44 (t, J = 7.4 Hz, 2H), 3.43 (t, J = 6.8 Hz, 4H), 3.55 (t, J = 4.8 Hz, 4H), 3.58 - 3.69 (m, 2H), 3.70 - 3.80 (m, 2H), 4.70 (t, J = 5.4 Hz, 1H).

[0503] Step 2: 4,4-bis(2-(octyloxy)ethoxy)butanoic acid

[0504] [ka]

[0505] From 4,4-bis(2-(octyloxy)ethoxy)butanenitrile according to general procedure B The yield was 136 mg (81%). 1 H NMR (400 MHz, DMSO-d6): δ 0.85 (t, J = 6.7 Hz, 6H), 1.07 - 1.36 (m, 20H), 1.47 (t, J = 6.9 Hz, 4H), 1.68 - 1.79 (m, 2H), 2.23 (t, J = 7.5 Hz, 2H), 3.36 (t, J = 6.5 Hz, 4H), 3.42 - 3.56 (m, 6H), 3. 58-3.63 (m, 2H), 4.55 (t, J = 5.6 Hz, 1H), 12.04 (s, 1H).

[0506] Step 3, 3-((4,4-bis(2-(octyloxy)ethoxy)butanoyl) Oxy)-2-(hydroxymethyl)propyl(9Z,12Z)-octadeca-9,12 -Dienoate

[0507] [ka]

[0508] Prepared from 4,4-bis(2-(octyloxy)ethoxy)butanoic acid according to general procedure C The yield was 147 mg (53%). 1 H NMR (400 MHz, chloroform-d): δ 0.83-0.92 (m, 9H), 1.13 - 1.41 (m, 37H), 1.55 - 1.65 (m, 3H), 1.92-2.09 (m, 6H), 2.13-2.2 2 (m, 1H), 2.27 - 2.36 (m, 3H), 2.42 (t, J = 7.4 Hz, 2H), 2.76 (t, J = 6.5 Hz, 2 H), 3.43 (t, J = 6.8 Hz, 4H), 3.55 (t, J = 4.8 Hz, 4H), 3.56 - 3.66 (m, 4H), 3.6 7 - 3.77 (m, 2H), 4.08 - 4.23 (m, 4H), 4.64 (t, J = 5.5 Hz, 1H), 5.28 - 5.41 (m, 4H).

[0509] Step 4: 3-((4,4-bis(2-(octyloxy)ethoxy)butanoyl) oxy)-2-((((3-(diethylamino)propoxy)carbonyl)oxy)methyl (9Z,12Z)-octadeca-9,12-dienoate (Example 96). 3-((4,4-bis(2-(octyloxy)ethoxy)butanoyl)oxy)-2- (Hydroxymethyl)propyl (9Z,12Z)-octadeca-9,12-dienoate and 3-(diethylamino)-1-propanol according to general procedure D. Amount 36 mg (62%). LCMS (Method B): (M+H) found m / z = 941.0, RT = 1.80 min.

[0510] [ka]

[0511] [Example 97] 3-((4,4-bis(2-(octyloxy)ethoxy)butanoyl)oxy)-2- (((((1-ethylpiperidin-3-yl)methoxy)carbonyl)oxy)methyl) Propyl (9Z,12Z)-octadeca-9,12-dienoate: 3-((4,4-biphenyl) 2-(hydroxymethyl)-2-(2-octyloxyethoxy)butanoyloxy)- ) propyl (9Z,12Z)-octadeca-9,12-dienoate and (1-ethyl Prepared from (piperidin-3-yl)methanol according to general procedure D. Yield 27 mg (5 8%). LCMS (Method B): (M+H) found m / z = 953.0, RT = 1.81 min.

[0512] [ka]

[0513] [Example 98] 3-((4,4-bis(((Z)-non-2-en-1-yl)oxy)butanoyl)o oxy)-2-((((3-(diethylamino)propoxy)carbonyl)oxy)methyl ) Propyl (9Z,12Z)-octadeca-9,12-dienoate

[0514] Step 1: 4,4-bis(((Z)-non-2-en-1-yl)oxy)butanediol Trill

[0515] [ka]

[0516] Prepared according to general procedure A using (Z)-non-2-en-1-ol. Yield 6 10mg (38%). 1 H NMR (400 MHz, chloroform-d) δ 0.87 (t, J = 6.7 Hz, 6H) , 1.19 - 1.41 (m, 18H), 1.97 (q, J = 7.3 Hz, 2H), 2.05 (q, J = 7.2 Hz, 4H), 2.42 (t, J = 7.4 Hz, 2H), 4.04 - 4.21 (m, 4H), 4.65 (t, J = 5.3 Hz, 1H), 5.45 - 5.65 (m, 4H).

[0517] Step 2: 4,4-bis(((Z)-non-2-en-1-yl)oxy)butanoic acid

[0518] [ka]

[0519] 4,4-bis(((Z)-non-2-en-1-yl)oxy)butanenitrile as a base Prepared according to procedure B. Yield 550 mg, 87%. 1 H NMR (400 MHz, DMSO-d6) δ 0 .85 (t, J = 6.4 Hz, 6H), 1.09 - 1.39 (m, 16H), 1.76 (q, J = 6.9 Hz, 2H), 2.02 (q , J = 6.8 Hz, 4H), 2.22 (t, J = 7.4 Hz, 2H), 3.95 - 4.11 (m, 4H), 4.53 (t, J = 5 .6 Hz, 1H), 5.41 - 5.57 (m, 4H), 12.06 (s, 1H).

[0520] Step 3: 3-((4,4-bis(((Z)-non-2-en-1-yl)oxy) Butanoyl)oxy)-2-(hydroxymethyl)propyl(9Z,12Z)-octadeca Carboxy-9,12-dienoate

[0521] [ka]

[0522] General Procedure from 4,4-bis(((Z)-non-2-en-1-yl)oxy)butanoic Acid Prepared according to C. Yield 390 mg, 71%. 1 H NMR (400 MHz, chloroform-d) δ 0.80 - 0.94 (m, 9H), 1.19 - 1.40 (m, 29H), 1.56 - 1.67 (m, 2H), 1.90 - 2.10 (m, 10H), 2.12 - 2.27 (m, 2H), 2.31 (t, J = 7.6 Hz, 2H), 2.41 (t, J = 7.4 Hz, 2H), 2 .76 (t, J = 6.4 Hz, 2H), 3.60 (t, J = 5.9 Hz, 2H), 4.01 - 4.23 (m, 8H), 4.58 (t, J = 5.5 Hz, 1H), 5.26 - 5.43 (m, 4H), 5.44 - 5.62 (m, 4H).

[0523] Step 4: 3-((4,4-bis(((Z)-non-2-en-1-yl)oxy) butanoyl)oxy)-2-((((3-(diethylamino)propoxy)carbonyl) Oxy)methyl)propyl (9Z,12Z)-octadeca-9,12-dienoate (actual Example 98) 3-((4,4-bis(((Z)-non-2-en-1-yl)oxy)butanol (9Z,12Z)-octadeca(2-hydroxymethyl)propyl 2-( ... The basic procedure is as follows: Prepared according to procedure D. Yield 70 mg (71%). LCMS (Method B): Found m / z for (M+H) = 8 76.9, RT = 1.81 minutes.

[0524] [ka]

[0525] [Example 99] 3-((4,4-bis(((Z)-non-2-en-1-yl)oxy)butanoyl)o oxy)-2-(((((1-ethylpiperidin-3-yl)methoxy)carbonyl)oxy 3-(methyl)propyl (9Z,12Z)-octadeca-9,12-dienoate (4,4-bis(((Z)-non-2-en-1-yl)oxy)butanoyl)oxy) -2-(hydroxymethyl)propyl(9Z,12Z)-octadeca-9,12-dieno Prepared from 1-ethylpiperidin-3-yl)methanol according to general procedure D. Yield: 45 mg (81%). LCMS (Method B): (M+H) found m / z = 860.9, RT = 1.5 6 minutes.

[0526] [ka]

[0527] [Example 100] 3-((4,4-bis(((Z)-hex-3-en-1-yl)oxy)butanoyl) oxy)-2-((((3-(diethylamino)propoxy)carbonyl)oxy)methyl (9Z,12Z)-octadeca-9,12-dienoate

[0528] Step 1: 4,4-bis(((Z)-hex-3-en-1-yl)oxy)butane Nitrile

[0529] [ka]

[0530] Prepared according to general procedure A using (Z)-hex-3-en-1-ol. 350mg (51%).

[0531] Step 2: 4,4-bis(((Z)-hex-3-en-1-yl)oxy)butane acid

[0532] [ka]

[0533] From 4,4-bis(((Z)-hex-3-en-1-yl)oxy)butanenitrile Prepared according to general procedure B. Yield 310 mg (82%). 1 H NMR (400 MHz, DMSO-d6) δ 0.92 (t, J = 7.5 Hz, 6H), 1.72 (q, J = 7.0 Hz, 2H), 1.95 - 2.07 (m, 4H), 2.17 - 2.27 (m, 6H), 3.31 - 3.42 (m, 2H), 3.44 - 3.55 (m, 2H), 4.49 (t, J = 5.6 Hz, 1H), 5.27 - 5.38 (m, 2H), 5.36 - 5.47 (m, 2H), 12.05 (s, 1H).

[0534] Step 3: 3-((4,4-bis(((Z)-hex-3-en-1-yl)oxy) )butanoyl)oxy)-2-(hydroxymethyl)propyl(9Z,12Z)-octa Deca-9,12-dienoate

[0535] [ka]

[0536] Basic synthesis from 4,4-bis(((Z)-hex-3-en-1-yl)oxy)butanoic acid Prepared according to procedure C. Yield 165 mg (37%). 1 H NMR (400 MHz, chloroform-d) δ 0.88 (t, J = 6.7 Hz, 3H), 0.95 (t, J = 7.5 Hz, 6H), 1.30 (s, 16H), 1.56 - 1. 66 (m, 2H), 1.88 - 1.98 (m, 2H), 2.04 (q, J = 7.2 Hz, 8H), 2.18 (t, J = 6.8 Hz, 2H), 2.31 (t, J = 7.3 Hz, 6H), 2.41 (t, J = 7.5 Hz, 2H), 2.76 (t, J = 6.4 Hz, 2H ), 3.42 (q, J = 7.4 Hz, 2H), 3.52 - 3.65 (m, 4H), 4.17 (t, J = 5.3 Hz, 4H), 4.52 (t, J = 5.6 Hz, 1H), 5.27 - 5.41 (m, 4H), 5.41 - 5.50 (m, 2H).

[0537] Step 4: 3-((4,4-bis(((Z)-hex-3-en-1-yl)oxy) )butanoyl)oxy)-2-((((3-(diethylamino)propoxy)carbonyl )oxy)methyl)propyl (9Z,12Z)-octadeca-9,12-dienoate ( Example 100) 3-((4,4-bis(((Z)-hex-3-en-1-yl)oxy) (9Z,12Z)-butanoyl)oxy)-2-(hydroxymethyl)propyl(9Z,12Z)-octanoyl From tadeca-9,12-dienoate and 3-(diethylamino)-1-propanol Prepared according to general procedure D. Yield 29 mg (52%). LCMS (Method B): Found m for (M+H). / z = ​​792.8, RT = 1.49 min.

[0538] [ka]

[0539] [Example 101] 3-((4,4-bis(((Z)-hex-3-en-1-yl)oxy)butanoyl) oxy)-2-(((((1-ethylpiperidin-3-yl)methoxy)carbonyl)o Oxy)methyl)propyl (9Z,12Z)-octadeca-9,12-dienoate: 3- ((4,4-bis(((Z)-hex-3-en-1-yl)oxy)butanoyl)oxy 9Z,12Z)-2-(hydroxymethyl)propyl octadeca-9,12-di From the enoate and (1-ethylpiperidin-3-yl)methanol according to general procedure D Yield 29 mg (51%). LCMS (Method B): Found m / z for (M+H) = 804.8, RT = 1.50 minutes.

[0540] [ka]

[0541] [Example 102] 3-((4,4-bis(((Z)-hept-3-en-1-yl)oxy)butanoyl) oxy)-2-((((3-(diethylamino)propoxy)carbonyl)oxy)methyl (9Z,12Z)-octadeca-9,12-dienoate

[0542] Step 1: 4,4-bis(((Z)-hept-3-en-1-yl)oxy)butane Nitrile

[0543] [ka]

[0544] Prepared according to general procedure A using (Z)-hept-3-en-1-ol. 310mg (41%). 1 H NMR (400 MHz, chloroform-d) δ 0.90 (t, J = 7.4 Hz, 6 H), 1.30 - 1.44 (m, 4H), 1.89 - 1.99 (m, 2H), 2.02 (q, J = 7.3 Hz, 4H), 2.32 (q, J = 7.0 Hz, 4H), 2.41 (t, J = 7.4 Hz, 2H), 3.40 - 3.48 (m, 2H), 3.54 - 3.66 (m, 2H), 4.58 (t, J = 5.3 Hz, 1H), 5.31 - 5.42 (m, 2H), 5.42 - 5.53 (m, 2H).

[0545] Step 2: 4,4-bis(((Z)-hept-3-en-1-yl)oxy)butane acid

[0546] [ka]

[0547] From 4,4-bis(((Z)-hept-3-en-1-yl)oxy)butanenitrile Prepared according to general procedure B. Yield 320 mg (83%). 1 H NMR (400 MHz, DMSO-d6) δ 0.86 (t, J = 7.3 Hz, 6H), 1.21 - 1.40 (m, 4H), 1.72 (q, J = 7.2 Hz, 2H), 1.99 (q, J = 6.9 Hz, 4H), 2.18 - 2.28 (m, 6H), 3.31 - 3.42 (m, 2H), 3.44 - 3.55 (m, 2H), 4.49 (t, J = 5.6 Hz, 1H), 5.31 - 5.47 (m, 4H), 12.05 (s, 1H).

[0548] Step 3: 3-((4,4-bis(((Z)-hept-3-en-1-yl)oxy) )butanoyl)oxy)-2-(hydroxymethyl)propyl(9Z,12Z)-octa Deca-9,12-dienoate

[0549] [ka]

[0550] Basic synthesis from 4,4-bis(((Z)-hept-3-en-1-yl)oxy)butanoic acid Prepared according to procedure C. Yield 295 mg, 45%. 1 H NMR (400 MHz, chloroform-d) δ 0.83 - 0.93 (m, 9H), 1.22 - 1.44 (m, 18H), 1.57 - 1.67 (m, 2H), 1.88 - 2.09 ( m, 10H), 2.19 (q, J = 5.9 Hz, 2H), 2.25 - 2.35 (m, 6H), 2.41 (t, J = 7.5 Hz, 2H) , 2.76 (t, J = 6.5 Hz, 2H), 3.36 - 3.47 (m, 2H), 3.52 - 3.65 (m, 4H), 4.10 - 4.2 3 (m, 4H), 4.52 (t, J = 5.5 Hz, 1H), 5.26 - 5.51 (m, 8H).

[0551] Step 4: 3-((4,4-bis(((Z)-hept-3-en-1-yl)oxy) )butanoyl)oxy)-2-((((3-(diethylamino)propoxy)carbonyl )oxy)methyl)propyl (9Z,12Z)-octadeca-9,12-dienoate ( Example 102) 3-((4,4-bis(((Z)-hept-3-en-1-yl)oxy) (9Z,12Z)-butanoyl)oxy)-2-(hydroxymethyl)propyl(9Z,12Z)-octanoyl From tadeca-9,12-dienoate and 3-(diethylamino)-1-propanol Prepared according to general procedure D. Yield 40 mg (72%). LCMS (Method B): Found m for (M+H). / z = ​​820.8, RT = 1.60 min.

[0552] [ka]

[0553] [Example 103] 3-((4,4-bis(((Z)-hept-3-en-1-yl)oxy)butanoyl) oxy)-2-(((((1-ethylpiperidin-3-yl)methoxy)carbonyl)o Oxy)methyl)propyl (9Z,12Z)-octadeca-9,12-dienoate: 3- ((4,4-bis(((Z)-hex-3-en-1-yl)oxy)butanoyl)oxy 9Z,12Z)-2-(hydroxymethyl)propyl octadeca-9,12-di From the enoate and (1-ethylpiperidin-3-yl)methanol according to general procedure D Yield 46 mg (83%). LCMS (Method B): (M+H) found m / z = 832.8, RT = 1.61 minutes.

[0554] [ka]

[0555] [Example 104] 3-((4,4-bis(oct-3-yn-1-yloxy)butanoyl)oxy)-2 -((((3-(diethylamino)propoxy)carbonyl)oxy)methyl)propyl (9Z,12Z)-Octadeca-9,12-dienoate

[0556] Step 1: 4,4-bis(oct-3-yn-1-yloxy)butanenitrile

[0557] [ka]

[0558] Prepared according to general procedure A using oct-3-yn-1-ol. Yield 410 ml g (28%). 1 H NMR (400 MHz, chloroform-d) δ 0.89 (t, J = 7.1 Hz, 6H), 1.31 - 1.51 (m, 8H), 1.92 - 2.02 (m, 2H), 2.09 - 2.19 (m, 4H), 2.38 - 2.49 (m, 6H), 3.50 - 3.61 (m, 2H), 3.64 - 3.74 (m, 2H), 4.67 (t, J = 5.4 Hz, 1H).

[0559] Step 2: 4,4-bis(oct-3-yn-1-yloxy)butanoic acid

[0560] [ka]

[0561] From 4,4-bis(oct-3-yn-1-yloxy)butanenitrile to General Procedure B Prepared as follows: Yield 310 mg (82%). 1 H NMR (400 MHz, DMSO-d6) δ 0.86 (t, J = 7.0 Hz, 6H), 1.26 - 1.45 (m, 8H), 1.74 (q, J = 7.0 Hz, 2H), 2.07 - 2.17 (m, 4H), 2.25 (t, J = 7.5 Hz, 2H), 2.30 - 2.41 (m, 4H), 3.40 - 3.51 (m, 2H), 3.52 - 3.62 (m, 2H), 4.56 (t, J = 5.7 Hz, 1H), 12.06 (s, 1H).

[0562] Step 3: 3-((4,4-bis(oct-3-yn-1-yloxy)butanoyl )oxy)-2-(hydroxymethyl)propyl(9Z,12Z)-octadeca-9,1 2-dienoate

[0563] [ka]

[0564] From 4,4-bis(oct-3-yn-1-yloxy)butanoic acid according to general procedure C Prepared. Yield: 390 mg (60%). 1 H NMR (400 MHz, chloroform-d) δ 0.83 - 0 .93 (m, 9H), 1.22 - 1.51 (m, 22H), 1.57 - 1.65 (m, 2H), 1.90 - 2.00 (m, 2H), 2.0 4 (q, J = 6.9 Hz, 4H), 2.08 - 2.23 (m, 6H), 2.31 (t, J = 7.6 Hz, 2H), 2.36 - 2.4 7 (m, 6H), 2.76 (t, J = 6.4 Hz, 2H), 3.48 - 3.71 (m, 6H), 4.08 - 4.23 (m, 4H), 4 .60 (t, J = 5.6 Hz, 1H), 5.26 - 5.42 (m, 4H).

[0565] Step 4: 3-((4,4-bis(oct-3-yn-1-yloxy)butanoyl )oxy)-2-((((3-(diethylamino)propoxy)carbonyl)oxy)methyl (9Z,12Z)-octadeca-9,12-dienoate (Example 104 ). 3-((4,4-bis(oct-3-yn-1-yloxy)butanoyl)oxy) -2-(hydroxymethyl)propyl(9Z,12Z)-octadeca-9,12-dieno Prepared according to general procedure D from 3-(diethylamino)-1-propanol and 3-(diethylamino)-1-propanol. Yield: 62 mg (67%). LCMS (Method B): Found m / z for (M+H) = 844.9, RT = 1.56 min.

[0566] [ka]

[0567] [Example 105] 3-((4,4-bis(oct-3-yn-1-yloxy)butanoyl)oxy)-2 -(((((1-ethylpiperidin-3-yl)methoxy)carbonyl)oxy)methyl ) Propyl (9Z,12Z)-octadeca-9,12-dienoate: 3-((4,4- Bis(((Z)-hex-3-en-1-yl)oxy)butanoyl)oxy)-2-( Hydroxymethyl)propyl (9Z,12Z)-octadeca-9,12-dienoate and and (1-ethylpiperidin-3-yl)methanol according to general procedure D. Yield 43 mg (51%). LCMS (Method B): (M+H) found m / z = 856.9, RT = 1.57 min.

[0568] [ka]

[0569] [Example 106] 3-((4,4-bis(((Z)-oct-3-en-1-yl)oxy)butanoyl) oxy)-2-((((3-(diethylamino)propoxy)carbonyl)oxy)methyl (9Z,12Z)-octadeca-9,12-dienoate

[0570] Step 1: 4,4-bis(((Z)-oct-3-en-1-yl)oxy)butane Nitrile

[0571] [ka]

[0572] Prepared according to general procedure A using (Z)-oct-3-en-1-ol. 100mg (40%). 1 H NMR (400 MHz, chloroform-d) δ 0.79 - 0.99 (m, 7H), 1 .16 - 1.39 (m, 9H), 1.90 - 1.98 (m, 2H), 2.03 (d, J = 6.9 Hz, 4H), 2.32 (q, J = 7.0 Hz, 4H), 2.41 (t, J = 7.4 Hz, 2H), 3.39 - 3.49 (m, 2H), 3.56 - 3.66 (m, 2H), 4.57 (t, J = 5.4 Hz, 1H), 5.29 - 5.40 (m, 2H), 5.41 - 5.52 (m, 2H).

[0573] Step 2: 4,4-bis(((Z)-oct-3-en-1-yl)oxy)butane acid

[0574] [ka]

[0575] From 4,4-bis(((Z)-oct-3-en-1-yl)oxy)butanenitrile Prepared according to general procedure B. Yield 320 mg, 83%. 1 H NMR (400 MHz, DMSO-d6) δ 0.83 - 0.89 (m, 6H), 1.18 - 1.36 (m, 8H), 1.72 (q, J = 7.0 Hz, 2H), 2.01 (q, J = 6.7 Hz, 4H), 2.15 - 2.29 (m, 6H), 3.31 - 3.42 (m, 2H), 3.44 - 3.54 (m, 2H), 4. 49 (t, J = 5.5 Hz, 1H), 5.30 - 5.47 (m, 4H), 12.06 (s, 1H).

[0576] Step 3: 3-((4,4-bis(((Z)-oct-3-en-1-yl)oxy) )butanoyl)oxy)-2-(hydroxymethyl)propyl(9Z,12Z)-octa Deca-9,12-dienoate

[0577] [ka]

[0578] Basic synthesis from 4,4-bis(((Z)-oct-3-en-1-yl)oxy)butanoic acid Prepared according to procedure C. Yield 310 mg, 48%. 1 H NMR (400 MHz, chloroform-d) δ 0.84 - 0.93 (m, 9H), 1.20 - 1.42 (m, 25H), 1.55 - 1.69 (m, 2H), 1.89 - 1.98 ( m, 2H), 1.98 - 2.08 (m, 7H), 2.13 - 2.22 (m, 2H), 2.31 (t, J = 7.2 Hz, 5H), 2.41 (t, J = 7.5 Hz, 2H), 2.76 (t, J = 6.0 Hz, 2H), 3.42 (q, J = 7.6 Hz, 2H), 3.52 - 3.65 (m, 4H), 4.09 - 4.28 (m, 4H), 4.48 - 4.56 (m, 1H), 5.27 - 5.41 (m, 5H), 5. 39 - 5.53 (m, 2H).

[0579] Step 4: 3-((4,4-bis(((Z)-oct-3-en-1-yl)oxy) )butanoyl)oxy)-2-((((3-(diethylamino)propoxy)carbonyl )oxy)methyl)propyl (9Z,12Z)-octadeca-9,12-dienoate ( Example 106) 3-((4,4-bis(((Z)-oct-3-en-1-yl)oxa (9Z,12Z)-butanoyl)oxy)-2-(hydroxymethyl)propyl(9Z,12Z)-octanoyl From tadeca-9,12-dienoate and 3-(diethylamino)-1-propanol Prepared according to general procedure D. Yield 47 mg (82%). LCMS (Method B): Found m for (M+H). / z = ​​848.9, RT = 1.72 min.

[0580] [ka]

[0581] [Example 107] 3-((4,4-bis(((Z)-oct-3-en-1-yl)oxy)butanoyl) oxy)-2-(((((1-ethylpiperidin-3-yl)methoxy)carbonyl)o Oxy)methyl)propyl (9Z,12Z)-octadeca-9,12-dienoate: 3- ((4,4-bis(((Z)-oct-3-en-1-yl)oxy)butanoyl)oxy 9Z,12Z)-2-(hydroxymethyl)propyl octadeca-9,12-di From the enoate and (1-ethylpiperidin-3-yl)methanol according to general procedure D Yield 45 mg (81%). LCMS (Method B): (M+H) found m / z = 860.9, RT = 1.56 minutes.

[0582] [ka]

[0583] [Example 108] 3-((4,4-bis(((Z)-non-6-en-1-yl)oxy)butanoyl)o oxy)-2-((((3-(diethylamino)propoxy)carbonyl)oxy)methyl ) Propyl (9Z,12Z)-octadeca-9,12-dienoate

[0584] Step 1: 4,4-bis(((Z)-non-6-en-1-yl)oxy)butanediol Trill

[0585] [ka]

[0586] Prepared according to general procedure A using (Z)-non-6-en-1-ol. Yield 6 10mg (37%). 1 H NMR (400 MHz, chloroform-d) δ 0.94 (t, J = 7.5 Hz, 6H) , 1.29 - 1.43 (m, 8H), 1.53 - 1.63 (m, 4H), 1.89 - 1.97 (m, 2H), 1.96 - 2.11 (m, 8H), 2.41 (t, J = 7.4 Hz, 2H), 3.36 - 3.47 (m, 2H), 3.53 - 3.65 (m, 2H), 4.54 ( t, J = 6.9 Hz, 1H), 5.18 - 5.52 (m, 4H).

[0587] Step 2: 4,4-bis(((Z)-non-6-en-1-yl)oxy)butanoic acid

[0588] [ka]

[0589] 4,4-bis(((Z)-non-6-en-1-yl)oxy)butanenitrile as a base Prepared according to procedure B. Yield 280 mg, 73%. 1 H NMR (400 MHz, DMSO-d6) δ 0 .91 (t, J = 7.5 Hz, 6H), 1.21 - 1.34 (m, 13H), 1.40 (t, J = 6.7 Hz, 2H), 1.43 - 1.52 (m, 2H), 1.72 (q, J = 7.1 Hz, 1H), 1.91 - 2.07 (m, 8H), 2.21 (t, J = 7.4 Hz , 1H), 3.32 - 3.41 (m, 3H), 3.42 - 3.53 (m, 1H), 4.32 (s, 1H), 5.24 - 5.39 (m, 4 H).

[0590] Step 3: 3-((4,4-bis(((Z)-non-6-en-1-yl)oxy) Butanoyl)oxy)-2-(hydroxymethyl)propyl(9Z,12Z)-octadeca Carboxy-9,12-dienoate

[0591] [ka]

[0592] General Procedure from 4,4-bis(((Z)-non-6-en-1-yl)oxy)butanoic Acid Prepared according to C. Yield 318 mg, 49%. 1 H NMR (400 MHz, chloroform-d) δ 0.86 - 0.90 (m, 4H), 0.94 (t, J = 7.5 Hz, 9H), 1.30 - 1.41 (m, 15H), 1.56 - 1.67 (m, 2H), 1.92 (q, J = 7.3 Hz, 3H), 2.01 - 2.04 (m, 5H), 2.19 (q, J = 6.5 Hz, 3H ), 2.31 (t, J = 7.6 Hz, 3H), 2.40 (t, J = 7.5 Hz, 3H), 2.76 (t, J = 6.4 Hz, 3H), 3.34 - 3.44 (m, 3H), 3.50 - 3.65 (m, 6H), 4.06 - 4.23 (m, 7H), 4.48 (t, J = 5.5 Hz, 2H), 5.17 - 5.50 (m, 12H).

[0593] Step 4: 3-((4,4-bis(((Z)-non-6-en-1-yl)oxy) butanoyl)oxy)-2-((((3-(diethylamino)propoxy)carbonyl) Oxy)methyl)propyl (9Z,12Z)-octadeca-9,12-dienoate (actual Example 108) 3-((4,4-bis(((Z)-non-6-en-1-yl)oxy) Butanoyl)oxy)-2-(hydroxymethyl)propyl(9Z,12Z)-octadeca Basic Synthesis from Benzene-9,12-dienoate and 3-(Diethylamino)-1-propanol Prepared according to procedure D. Yield 36 mg (69%). LCMS (Method B): Found m / z for (M+H) = 876.9, RT = 1.78 minutes.

[0594] [ka]

[0595] [Example 109] 3-((4,4-bis(((Z)-non-6-en-1-yl)oxy)butanoyl)o oxy)-2-(((((1-ethylpiperidin-3-yl)methoxy)carbonyl)oxy 3-(methyl)propyl (9Z,12Z)-octadeca-9,12-dienoate (4,4-bis(((Z)-non-6-en-1-yl)oxy)butanoyl)oxy) -2-(hydroxymethyl)propyl(9Z,12Z)-octadeca-9,12-dieno Prepared from 1-ethylpiperidin-3-yl)methanol according to general procedure D. Yield: 37 mg (67%). LCMS (Method B): (M+H) found m / z = 888.9, RT = 1.7 7 minutes.

[0596] [ka]

[0597] [Example 110] 3-((4,4-bis((8-fluorooctyl)oxy)butanoyl)oxy)-2- ((((3-(diethylamino)propoxy)carbonyl)oxy)methyl)propyl( 9Z,12Z)-Octadeca-9,12-dienoate

[0598] Step 1: 4,4-bis((8-bromooctyl)oxy)butanenitrile

[0599] [ka]

[0600] Prepared according to general procedure A using 8-bromootan-1-ol. Yield 750 ml g (33%). 1 H NMR (400 MHz, chloroform-d) δ 1.22 - 1.41 (m, 12H), 1.37 - 1 .48 (m, 4H), 1.56 (t, J = 7.3 Hz, 4H), 1.78 - 1.91 (m, 4H), 1.88 - 1.98 (m, 2H), 2.41 (t, J = 7.3 Hz, 2H), 3.33 - 3.47 (m, 6H), 3.51 - 3.64 (m, 2H), 4.54 (t, J = 5.3 Hz, 1H).

[0601] Step 2: 4,4-bis((8-fluorooctyl)oxy)butanenitrile

[0602] [ka]

[0603] Tetrabutyl fluoride in THF (1 M, 4.65 mL, 4.65 mmol, 3 Eq) A solution of ammonium was dissolved in 4,4-bis((8-bromooctyl)oxy)butanenitrile ( The mixture was stirred at 80°C for 24 hours. After this time, the reaction mass was rinsed with ice-cold water and then the organic layer was extracted with ethyl acetate (30 mL x 3). The organic layer was washed with brine (5 mL x 3) and then dried over sodium sulfate. The organic layer was evaporated to give the crude product, which was purified by column chromatography (ethyl acetate / hexane) and 4,4-bis((8-fluorooctyl)oxy)butadiene This gave benzonitrile (210 mg, 37%) as a pale yellow gum. 1 H NMR (400 MHz, Chloroform-d) δ 1.30 - 1.45 (m, 18H), 1.56 - 1.77 (m, 6H), 1.88 - 1.98 (m, 2H ), 2.41 (t, J = 7.3 Hz, 2H), 3.32 - 3.47 (m, 2H), 3.54 - 3.64 (m, 2H), 4.37 (t, J = 6.2 Hz, 2H), 4.48 (t, J = 6.1 Hz, 2H), 4.54 (t, J = 5.3 Hz, 1H).

[0604] Step 3: 4,4-bis((8-fluorooctyl)oxy)butanoic acid

[0605] [ka]

[0606] From 4,4-bis((8-fluorooctyl)oxy)butanenitrile according to general procedure B The yield was 200 mg (92%). 1 H NMR (400 MHz, DMSO-d6) δ 1.21 - 1.37 (m, 14H), 1.48 (t, J = 6.6 Hz, 4H), 1.53 - 1.77 (m, 6H), 2.22 (t, J = 7.4 Hz, 2 H), 3.27 - 3.40 (m, 3H), 3.43 - 3.54 (m, 2H), 4.36 (t, J = 6.1 Hz, 2H), 4.41 - 4 .52 (m, 3H), 12.05 (s, 1H).

[0607] Step 4: 3-((4,4-bis((8-fluorooctyl)oxy)butanoyl) Oxy)-2-(hydroxymethyl)propyl(9Z,12Z)-octadeca-9,12 -Dienoate

[0608] [ka]

[0609] Prepared from 4,4-bis((8-fluorooctyl)oxy)butanoic acid according to general procedure C The yield was 160 mg, 37%. 1 H NMR (400 MHz, chloroform-d) δ 0.88 (t, J = 6.9 Hz, 3H), 1.21 - 1.44 (m, 34H), 1.58 - 1.77 (m, 6H), 1.87 - 1.97 (m, 2H), 2. 04 (q, J = 6.8 Hz, 4H), 2.13 - 2.24 (m, 2H), 2.31 (t, J = 7.6 Hz, 2H), 2.40 (t, J = 7.5 Hz, 2H), 2.76 (t, J = 6.4 Hz, 2H), 3.34 - 3.44 (m, 2H), 3.50 - 3.65 (m, 4H), 4.10 - 4.23 (m, 4H), 4.36 (t, J = 6.1 Hz, 2H), 4.44 - 4.52 (m, 3H), 5.25 - 5.44 (m, 4H).

[0610] Step 5: 3-((4,4-bis((8-fluorooctyl)oxy)butanoyl) oxy)-2-((((3-(diethylamino)propoxy)carbonyl)oxy)methyl (9Z,12Z)-octadeca-9,12-dienoate (Example 110) 3-((4,4-bis((8-fluorooctyl)oxy)butanoyl)oxy)-2 -(hydroxymethyl)propyl(9Z,12Z)-octadeca-9,12-dienoate Prepared according to general procedure D from methylpropanol and 3-(diethylamino)-1-propanol. Yield 18 mg (21%). LCMS (Method B): (M+H) found m / z = 888.9, RT = 1.40 min.

[0611] [ka]

[0612] [Example 111] 3-((4,4-bis((8-fluorooctyl)oxy)butanoyl)oxy)-2- (((((1-ethylpiperidin-3-yl)methoxy)carbonyl)oxy)methyl) Propyl (9Z,12Z)-octadeca-9,12-dienoate: 3-((4,4-biphenyl) bis((8-fluorooctyl)oxy)butanoyl)oxy)-2-(hydroxymethyl ) propyl (9Z,12Z)-octadeca-9,12-dienoate and (1-ethyl Prepared from (piperidin-3-yl)methanol according to general procedure D. Yield 41 mg (4 8%). LCMS (Method B): (M+H) found m / z = 900.9, RT = 1.41 min.

[0613] [ka]

[0614] [Example 112] 3-((6,6-bis(((Z)-oct-3-en-1-yl)oxy)hexanoyl )oxy)-2-((((3-(diethylamino)propoxy)carbonyl)oxy)methyl (9Z,12Z)-octadeca-9,12-dienoate

[0615] Step 1: 2-Methoxycyclohexan-1-one oxime

[0616] [ka]

[0617] 2-Methoxycyclohexan-1-one in MeOH-H2O (150 ml, 1:2) (5 g, 39.0 mmol) was added to a stirred solution of sodium acetate (6.41 g, 78.1 mmol) ol) was added, followed by hydroxylamine hydrochloride (5.43 g, 78.1 mmol). The resulting mixture was heated in a sealed tube at 70° C. for 16 hours. After that time, M The eOH was evaporated and the mixture was diluted with water, extracted with ethyl acetate (3 x 50 mL) and brine. The crude product thus obtained was washed with water, dried over sodium sulfate, and concentrated under reduced pressure. The product was purified by flash column chromatography eluting with 3% ethyl acetate in hexane. and purified to give 2-methoxycyclohexan-1-one oxime (4.1 g, 88%). Obtained as a colored oil. 1 H NMR (400 MHz, DMSO-d6) δ 1.17-1.32 (m, 1H), 1.40-1.56 (m , 2H), 1.59-1.82 (m, 3H), 1.92-1.99 (m, 1H), 2.93-3.01 (m, 1H), 3.11 (s, 3H), 3. 63 (s, 1H), 10.61 (s, 1H).

[0618] Step 2: 6,6-Dimethoxyhexanenitrile

[0619] [ka]

[0620] 2-Methoxycyclohexan-1-one oxime (2.0 g) in CCl4 (20 ml) To a stirred solution of thionyl chloride (1.2 ml, 16.8 mmol) was added Add dropwise at 0°C and stir for 10 minutes. After that time, add dry MeOH (20 ml) The reaction mixture was added at ~10°C. After the addition was complete, the reaction mixture was allowed to reach room temperature and was then stirred for 2 hours. The mixture was then diluted with ethyl acetate, washed with brine, and After drying and concentration, 6,6-dimethoxyhexanenitrile (1.6 g, 81%) was obtained as a pale yellow solid. Obtained as an oil. 1 H NMR (400 MHz, chloroform-d) δ 1.44 - 1.56 (m, 2H), 1.57 - 1.75 (m, 4H), 2.34 (t, J = 7.0 Hz, 2H), 3.31 (s, 6H), 4.34 (t, J = 5.5 Hz, 1H) .

[0621] Step 3: 6,6-bis(((Z)-oct-3-en-1-yl)oxy)hexa Nitrile

[0622] [ka]

[0623] From 6,6-dimethoxyhexanenitrile and (Z)-oct-3-en-1-ol Prepared according to general procedure A from . Yield 280 mg (50%). 1 H NMR (400 MHz, DMSO-d6 ) δ 0.83 - 0.89 (m, 6H), 1.21 - 1.32 (m, 8H), 1.31 - 1.43 (m, 2H), 1.47 - 1.61 (m, 4H), 2.01 (q, J = 6.7 Hz, 4H), 2.23 (q, J = 6.8 Hz, 4H), 2.47 (d, J = 7.1 Hz , 2H), 3.32 - 3.42 (m, 2H), 3.44 - 3.54 (m, 2H), 4.47 (t, J = 5.6 Hz, 1H), 5.30 - 5.47 (m, 4H).

[0624] Step 4: 6,6-bis(((Z)-oct-3-en-1-yl)oxy)hexa Phosphoric acid

[0625] [ka]

[0626] Prepared from 6,6-dimethoxyhexanenitrile according to general procedure B. Yield 260 ml g(80%). 1 H NMR (400 MHz, DMSO-d6) δ 0.86 (q, J = 3.8, 5.1 Hz, 6H), 1.21 - 1.34 (m, 10H), 1.48 (t, J = 7.6 Hz, 4H), 2.00 (t, J = 6.7 Hz, 4H), 2.14 - 2.27 (m, 6H), 3.30 - 3.41 (m, 2H), 3.42 - 3.53 (m, 2H), 4.44 (t, J = 5.6 Hz, 1H), 5.3 0 - 5.46 (m, 4H), 11.97 (s, 1H).

[0627] Step 5: 3-((6,6-bis(((Z)-oct-3-en-1-yl)oxy) )hexanoyl)oxy)-2-(hydroxymethyl)propyl(9Z,12Z)-octanoyl Tadeca-9,12-dienoate

[0628] [ka]

[0629] 6,6-bis(((Z)-oct-3-en-1-yl)oxy)hexanoic acid as a base Prepared according to procedure C. Yield 70 mg (55%). 1 H NMR (400 MHz, chloroform-d) δ 0.83 - 0.95 (m, 9H), 1.17 - 1.42 (m, 25H), 1.57 - 1.70 (m, 5H), 2.04 (d, J = 7.0 Hz, 8H), 2.18 (t, J = 6.2 Hz, 2H), 2.31 (t, J = 6.7 Hz, 8H), 2.76 (t, J = 6 .5 Hz, 2H), 3.41 (q, J = 7.5 Hz, 2H), 3.50 - 3.64 (m, 4H), 4.09 - 4.20 (m, 4H), 4.48 (t, J = 5.6 Hz, 1H), 5.32 - 5.37 (m, 5H), 5.33 - 5.48 (m, 3H).

[0630] Step 6: 3-((6,6-bis(((Z)-oct-3-en-1-yl)oxy) )hexanoyl)oxy)-2-((((3-(diethylamino)propoxy)carbonyl (9Z,12Z)-octadeca-9,12-dienoate (Example 112) 3-((6,6-bis(((Z)-oct-3-en-1-yl)o 2-(hydroxymethyl)propyl(9Z,12Z)-(hydroxy)hexanoyl)oxy Octadeca-9,12-dienoate and 3-(diethylamino)-1-propanol Prepared from according to general procedure D. Yield 37 mg (64%). LCMS (Method B): (M+H) Measured value m / z = 877.0, RT = 1.70 minutes.

[0631] [ka]

[0632] [Example 113] 3-((6,6-bis(((Z)-oct-3-en-1-yl)oxy)hexanoyl )oxy)-2-(((((1-ethylpiperidin-3-yl)methoxy)carbonyl) Oxy)methyl)propyl (9Z,12Z)-octadeca-9,12-dienoate: 3 -((6,6-bis(((Z)-oct-3-en-1-yl)oxy)hexanoyl) Oxy)-2-(hydroxymethyl)propyl(9Z,12Z)-octadeca-9,12 -dienoate and (1-ethylpiperidin-3-yl)methanol to general procedure D Prepared as follows: Yield 89 mg (70%). LCMS (Method B): (M+H) found m / z = 889.0. RT = 1.79 minutes.

[0633] [ka]

[0634] [Example 114] 3-((6,6-bis(((Z)-hex-3-en-1-yl)oxy)hexanoyl )oxy)-2-((((3-(diethylamino)propoxy)carbonyl)oxy)methyl (9Z,12Z)-octadeca-9,12-dienoate

[0635] Step 1: 6,6-bis(((Z)-hex-3-en-1-yl)oxy)hexa Nitrile

[0636] [ka]

[0637] From 6,6-dimethoxyhexanenitrile and (Z)-hex-3-en-1-ol Prepared according to general procedure A from Isaac et al. Yield 190 mg, 42%. 1 H NMR (400 MHz, chloroform) Lum-d) δ 0.96 (t, J = 7.5 Hz, 6H), 1.58 - 1.74 (m, 6H), 1.99 - 2.11 (m, 4H), 2 .20 - 2.38 (m, 6H), 3.36 - 3.47 (m, 2H), 3.52 - 3.62 (m, 2H), 4.48 (t, J = 5.5 H z, 1H), 5.27 - 5.39 (m, 2H), 5.46 (q, J = 7.6 Hz, 2H).

[0638] Step 2: 6,6-bis(((Z)-hex-3-en-1-yl)oxy)hexa Phosphoric acid

[0639] [ka]

[0640] 6,6-bis(((Z)-hex-3-en-1-yl)oxy)hexanenitrile Prepared according to general procedure B from Isaac et al. Yield 170 mg, 82%. 1 H NMR (400 MHz, DMSO-d6) δ 0.92 (t, J = 7.5 Hz, 6H), 1.21 - 1.32 (m, 2H), 1.42 - 1.55 (m, 4H), 1.95 - 2. 07 (m, 4H), 2.14 - 2.27 (m, 6H), 3.31 - 3.42 (m, 2H), 3.43 - 3.54 (m, 2H), 4.45 (t, J = 5.6 Hz, 1H), 5.27 - 5.38 (m, 2H), 5.36 - 5.47 (m, 2H), 11.96 (s, 1H).

[0641] Step 3: 3-((6,6-bis(((Z)-hex-3-en-1-yl)oxy) )hexanoyl)oxy)-2-(hydroxymethyl)propyl(9Z,12Z)-octanoyl Tadeca-9,12-dienoate

[0642] [ka]

[0643] 6,6-bis(((Z)-hex-3-en-1-yl)oxy)hexanoic acid as a base Prepared according to procedure C. Yield 60 mg (45%). 1 H NMR (400 MHz, chloroform-d) δ 0.88 (t, J = 6.7 Hz, 3H), 0.95 (t, J = 7.5 Hz, 6H), 1.18 - 1.44 (m, 15H), 1. 56 - 1.70 (m, 7H), 1.99 - 2.11 (m, 8H), 2.13 - 2.23 (m, 2H), 2.23 - 2.37 (m, 8H) , 2.76 (t, J = 6.4 Hz, 2H), 3.36-3.47 (m, 2H), 3.50 - 3.64 (m, 4H), 4.09 - 4.24 (m, 4H), 4.48 (t, J = 5.6 Hz, 1H), 5.26 - 5.51 (m, 8H).

[0644] Step 4: 3-((6,6-bis(((Z)-hex-3-en-1-yl)oxy) )hexanoyl)oxy)-2-((((3-(diethylamino)propoxy)carbonyl (9Z,12Z)-octadeca-9,12-dienoate (Example 114) 3-((6,6-bis(((Z)-hex-3-en-1-yl)o 2-(hydroxymethyl)propyl(9Z,12Z)-(hydroxy)hexanoyl)oxy Octadeca-9,12-dienoate and 3-(diethylamino)-1-propanol Prepared from according to general procedure D. Yield 36 mg (62%). LCMS (Method B): (M+H) Measured m / z = 820.9, RT = 1.70 minutes.

[0645] [ka]

[0646] [Example 115] 3-((6,6-bis(((Z)-hex-3-en-1-yl)oxy)hexanoyl )oxy)-2-(((((1-ethylpiperidin-3-yl)methoxy)carbonyl) Oxy)methyl)propyl (9Z,12Z)-octadeca-9,12-dienoate: 3 -((6,6-bis(((Z)-hex-3-en-1-yl)oxy)hexanoyl) Oxy)-2-(hydroxymethyl)propyl(9Z,12Z)-octadeca-9,12 -dienoate and (1-ethylpiperidin-3-yl)methanol to general procedure D Prepared as follows: Yield 69 mg (72%). LCMS (Method B): (M+H) found m / z = 833.0. RT = 1.60 minutes.

[0647] [ka]

[0648] [Example 116] 3-((6,6-bis(((Z)-oct-5-en-1-yl)oxy)hexanoyl )oxy)-2-((((3-(diethylamino)propoxy)carbonyl)oxy)methyl (9Z,12Z)-octadeca-9,12-dienoate

[0649] Step 1: 6,6-bis(((Z)-oct-5-en-1-yl)oxy)hexa Nitrile

[0650] [ka]

[0651] From 6,6-dimethoxyhexanenitrile and (Z)-oct-5-en-1-ol Prepared according to general procedure A from Sigma-Aldrich. Yield 210 mg (47%). 1 H NMR (400 MHz, chloro Holm-d) δ 0.94 (t, J = 7.5 Hz, 6H), 1.34 - 1.52 (m, 4H), 1.47 - 1.74 (m, 10H) , 1.95 - 2.10 (m, 8H), 2.33 (t, J = 7.1 Hz, 2H), 3.34 - 3.45 (m, 2H), 3.51 - 3.6 1 (m, 2H), 4.44 (t, J = 5.5 Hz, 1H), 5.25 - 5.42 (m, 4H).

[0652] Step 2: 6,6-bis(((Z)-oct-5-en-1-yl)oxy)hexa Phosphoric acid

[0653] [ka]

[0654] 6,6-bis(((Z)-oct-5-en-1-yl)oxy)hexanenitrile Prepared according to general procedure B from Isaac et al. Yield 150 mg, 94%. 1 H NMR (400 MHz, DMSO-d6) δ 0.91 (t, J = 7.5 Hz, 6H), 1.24 - 1.41 (m, 6H), 1.42 - 1.53 (m, 8H), 1.93 - 2. 06 (m, 8H), 2.18 (t, J = 7.3 Hz, 2H), 3.31 - 3.40 (m, 2H), 3.43 - 3.53 (m, 2H), 4.40 (t, J = 5.6 Hz, 1H), 5.19 - 5.45 (m, 4H), 11.97 (s, 1H).

[0655] Step 3: 3-((6,6-bis(((Z)-oct-5-en-1-yl)oxy) )hexanoyl)oxy)-2-(hydroxymethyl)propyl(9Z,12Z)-octanoyl Tadeca-9,12-dienoate

[0656] [ka]

[0657] 6,6-bis(((Z)-oct-5-en-1-yl)oxy)hexanoic acid as a base Prepared according to procedure C. Yield 90 mg (45%). 1 H NMR (400 MHz, chloroform-d) δ 0.88 (t, J = 6.8 Hz, 3H), 0.94 (t, J = 7.5 Hz, 6H), 1.20 - 1.47 (m, 23H), 1. 50 - 1.70 (m, 9H), 1.94 - 2.11 (m, 12H), 2.10 - 2.23 (m, 2H), 2.27 - 2.37 (m, 4H) ), 2.76 (t, J = 6.4 Hz, 2H), 3.34 - 3.44 (m, 2H), 3.50 - 3.58 (m, 2H), 3.60 (t, J = 5.6 Hz, 2H), 4.09 - 4.24 (m, 4H), 4.44 (t, J = 5.6 Hz, 1H), 5.25 - 5.43 (m, 8H).

[0658] Step 4: 3-((6,6-bis(((Z)-oct-5-en-1-yl)oxy) )hexanoyl)oxy)-2-((((3-(diethylamino)propoxy)carbonyl (9Z,12Z)-octadeca-9,12-dienoate (Example 116) 3-((6,6-bis(((Z)-oct-5-en-1-yl)o 2-(hydroxymethyl)propyl(9Z,12Z)-(hydroxy)hexanoyl)oxy Octadeca-9,12-dienoate and 3-(diethylamino)-1-propanol Prepared from according to general procedure D. Yield 24 mg (57%). LCMS (Method B): (M+H) Measured m / z = 876.8, RT = 1.70 minutes.

[0659] [ka]

[0660] [Example 117] 3-((6,6-bis(((Z)-oct-5-en-1-yl)oxy)hexanoyl )oxy)-2-(((((1-ethylpiperidin-3-yl)methoxy)carbonyl) Oxy)methyl)propyl (9Z,12Z)-octadeca-9,12-dienoate: 3 -((6,6-bis(((Z)-oct-5-en-1-yl)oxy)hexanoyl) Oxy)-2-(hydroxymethyl)propyl(9Z,12Z)-octadeca-9,12 -dienoate and (1-ethylpiperidin-3-yl)methanol to general procedure D Prepared as follows. Yield 50 mg (50%). LCMS (Method B): (M+H) found m / z = 889.2, RT = 1.73 minutes.

[0661] [ka]

[0662] [Example 118] 3-((6,6-bis(((Z)-dec-4-en-1-yl)oxy)hexanoyl) oxy)-2-((((3-(diethylamino)propoxy)carbonyl)oxy)methyl (9Z,12Z)-octadeca-9,12-dienoate

[0663] Step 1: 6,6-bis(((Z)-dec-4-en-1-yl)oxy)hexane Nitrile

[0664] [ka]

[0665] From 6,6-dimethoxyhexanenitrile and (Z)-dec-4-en-1-ol Prepared according to general procedure A. Yield 150 mg (38%). 1 H NMR (400 MHz, chloroform) Lum-d) δ 0.88 (t, J = 6.7 Hz, 6H), 1.17 - 1.41 (m, 12H), 1.45 - 1.57 (m, 2H), 1.59 - 1.74 (m, 8H), 1.87 - 2.20 (m, 8H), 2.33 (t, J = 7.1 Hz, 2H), 3.35 - 3.45 (m, 2H), 3.50 - 3.70 (m, 2H), 4.44 (t, J = 5.5 Hz, 1H), 5.28 - 5.45 (m, 4H)

[0666] Step 2: 6,6-bis(((Z)-dec-4-en-1-yl)oxy)hexane acid

[0667] [ka]

[0668] From 6,6-bis(((Z)-dec-4-en-1-yl)oxy)hexanenitrile Prepared according to general procedure B. Yield 150 mg, 95%. 1 H NMR (400 MHz, DMSO-d6) δ 0.85 (t, J = 6.7 Hz, 6H), 1.18 - 1.37 (m, 14H), 1.45 - 1.56 (m, 8H), 1.91 - 2.1 0 (m, 8H), 2.18 (t, J = 7.4 Hz, 2H), 3.32 - 3.41 (m, 2H), 3.43 - 3.53 (m, 2H), 4 .37 - 4.44 (m, 1H), 5.25 - 5.46 (m, 4H), 11.96 (s, 1H).

[0669] Step 3: 3-((6,6-bis(((Z)-dec-4-en-1-yl)oxy) Hexanoyl)oxy)-2-(hydroxymethyl)propyl(9Z,12Z)-octa Deca-9,12-dienoate

[0670] [ka]

[0671] Basic synthesis from 6,6-bis(((Z)-dec-4-en-1-yl)oxy)hexanoic acid Prepared according to procedure C. Yield 87 mg (42%). 1 H NMR (400 MHz, chloroform-d) δ 0.88 (t, J = 6.8 Hz, 9H), 1.17 - 1.45 (m, 24H), 1.51 - 1.71 (m, 10H), 1.96 - 2.17 (m, 12H), 2.32 (q, J = 7.3 Hz, 4H), 2.76 (t, J = 6.5 Hz, 2H), 3.35 - 3.45 ( m, 2H), 3.51 - 3.64 (m, 4H), 4.09 - 4.24 (m, 4H), 4.44 (t, J = 5.6 Hz, 1H), 5.28 - 5.44 (m, 8H).

[0672] Step 4: 3-((6,6-bis(((Z)-dec-4-en-1-yl)oxy) hexanoyl)oxy)-2-((((3-(diethylamino)propoxy)carbonyl )oxy)methyl)propyl (9Z,12Z)-octadeca-9,12-dienoate. 3-((6,6-bis(((Z)-dec-4-en-1-yl)oxy)hexanoyl) Oxy)-2-(hydroxymethyl)propyl(9Z,12Z)-octadeca-9,12 -dienoate and 3-(diethylamino)-1-propanol according to general procedure D Yield 32 mg (59%). LCMS (Method B): (M+H) found m / z = 932.9, RT = 1.70 minutes.

[0673] [ka]

[0674] [Example 119] 3-((6,6-bis(((Z)-dec-4-en-1-yl)oxy)hexanoyl) oxy)-2-(((((1-ethylpiperidin-3-yl)methoxy)carbonyl)o Oxy)methyl)propyl (9Z,12Z)-octadeca-9,12-dienoate: 3- ((6,6-bis(((Z)-dec-4-en-1-yl)oxy)hexanoyl)oxy 9Z,12Z)-2-(hydroxymethyl)propyl octadeca-9,12-di From the enoate and (1-ethylpiperidin-3-yl)methanol according to general procedure D Yield 51 mg (52%). LCMS (Method B): (M+H) found m / z = 945.2, RT = 1.70 minutes.

[0675] [ka]

[0676] [Example 120] 3-((6,6-bis((3,7-dimethyloct-6-en-1-yl)oxy)hexyl (((3-(diethylamino)propoxy)carbonyl)oxy (9Z,12Z)-Octadeca-9,12-dienoate

[0677] Step 1: 6,6-bis((3,7-dimethyloct-6-en-1-yl)oxy) ) Hexanenitrile

[0678] [ka]

[0679] 6,6-Dimethoxyhexanenitrile and 3,7-dimethyloct-6-ene-1- Prepared from ol according to general procedure A. Yield 220 mg (42%). 1 H NMR (400 MHz, Chloroform-d) δ 0.88 (d, J = 6.4 Hz, 6H), 1.11 - 1.22 (m, 3H), 1.25 - 1.43 ( m, 6H), 1.45 - 1.77 (m, 19H), 1.87 - 2.04 (m, 4H), 2.33 (t, J = 7.1 Hz, 2H), 3.3 5 - 3.50 (m, 2H), 3.52 - 3.66 (m, 2H), 4.44 (t, J = 5.5 Hz, 1H), 5.08 (t, J = 7. 3 Hz, 2H).

[0680] Step 2: 6,6-bis((3,7-dimethyloct-6-en-1-yl)oxy) ) Hexanoic acid

[0681] [ka]

[0682] 6,6-bis((3,7-dimethyloct-6-en-1-yl)oxy)hexanedi Prepared from tolyl according to general procedure B. Yield 220 mg (95%). 1 H NMR (400 MHz, DMSO-d6) δ 0.85 (d, J = 5.9 Hz, 6H), 1.13 - 1.03 (m, 2H), 1.40 - 1.17 (m, 8H), 1.58 - 1.44 (m, 12H), 1.70 - 1.59 (m, 6H), 2.03 - 1.87 (m, 5H), 2.18 (t, J = 7. 3 Hz, 1H), 3.44 - 3.33 (m, 2H), 3.60 - 3.44 (m, 2H), 4.40 (t, J = 5.4 Hz, 1H), 5 .07 (t, J = 7.3 Hz, 2H), 11.97 (s, 1H).

[0683] Step 3: 3-((6,6-bis((3,7-dimethyloct-6-en-1-yl) )oxy)hexanoyl)oxy)-2-(hydroxymethyl)propyl(9Z,12Z )-Octadeca-9,12-dienoate

[0684] [ka]

[0685] 6,6-bis((3,7-dimethyloct-6-en-1-yl)oxy)hexanoic acid Prepared from according to general procedure C. Yield 90 mg (42%). 1 H NMR (400 MHz, chloro Holm-d) δ 0.88 (d, J = 6.1 Hz, 10H), 1.18 - 1.45 (m, 15H), 1.51 - 1.57 (m, 22 H), 1.67 (s, 5H), 1.83 - 2.09 (m, 8H), 2.16 - 2.23 (m, 2H), 2.32 (q, J = 7.5 Hz, 4H), 2.75 (d, J = 6.1 Hz, 2H), 3.32 - 3.69 (m, 7H), 4.10 - 4.21 (m, 4H), 4.40 - 4.48 (m, 1H), 5.03 - 5.16 (m, 2H), 5.28 - 5.42 (m, 4H).

[0686] Step 4: 3-((6,6-bis((3,7-dimethyloct-6-en-1-yl) )oxy)hexanoyl)oxy)-2-((((3-(diethylamino)propoxy) Carbonyl)oxy)methyl)propyl(9Z,12Z)-octadeca-9,12-dienes 3-((6,6-bis((3,7-dimethylocta-6-enoate)) (hydroxymethyl)propyl (hydroxymethyl)-2-(hydroxymethyl)propyl (hydroxymethyl)-1-(hydroxymethyl)propyl (hydroxymethyl)-2-(hydroxymethyl)propyl ( ... 9Z,12Z)-Octadeca-9,12-dienoate and 3-(diethylamino)- Prepared from 1-propanol according to general procedure D. Yield 32 mg (59%). LCMS (method Method B): (M+H) measured m / z = 933.0, RT = 1.70 min.

[0687] [ka]

[0688] [Example 121] 3-((6,6-bis((3,7-dimethyloct-6-en-1-yl)oxy)hexyl ((((1-ethylpiperidin-3-yl)methoxy)-2-(((((1-ethylpiperidin-3-yl)oxy)-2-(((((1-ethylpiperidin-3-yl)methoxy ... (9Z,12Z)-octadeca-9,12-dienoic acid, (9Z,12Z)-(methyl)propyl 3-((6,6-bis((3,7-dimethyloct-6-en-1-yl)oxy)- ... )hexanoyl)oxy)-2-(hydroxymethyl)propyl(9Z,12Z)-octanoyl Tadeca-9,12-dienoate and (1-ethylpiperidin-3-yl)methanol Prepared from according to general procedure D. Yield 32 mg (59%). LCMS (Method B): (M+H) Measured value m / z = 945.2, RT = 1.96 minutes.

[0689] [ka]

[0690] [Example 122] 3-((6,6-bis((7,7,8,8,8-pentafluorooctyl)oxy)hexyl (((3-(diethylamino)propoxy)carbonyl)oxy (9Z,12Z)-Octadeca-9,12-dienoate

[0691] Step 1: 6,6-bis((7,7,8,8,8-pentafluorooctyl)oxy) ) Hexanenitrile

[0692] [ka]

[0693] 6,6-Dimethoxyhexanenitrile and 7,7,8,8,8-Pentafluorooctyl Prepared from tan-1-ol according to general procedure A. Yield 300 mg (44%). 1 H NMR (400 MHz, chloroform-d) δ 1.35 - 1.45 (m, 8H), 1.45 - 1.56 (m, 2H), 1.54 - 1 .74 (m, 12H), 1.91 - 2.09 (m, 4H), 2.34 (t, J = 7.0 Hz, 2H), 3.34 - 3.45 (m, 2H) , 3.51 - 3.61 (m, 2H), 4.44 (t, J = 5.5 Hz, 1H)

[0694] Step 2: 6,6-bis((7,7,8,8,8-pentafluorooctyl)oxy) ) Hexanoic acid

[0695] [ka]

[0696] 6,6-bis((7,7,8,8,8-pentafluorooctyl)oxy)hexanedi Prepared from tolyl according to general procedure B. Yield 296 mg (95%). 1 H NMR (400 MHz, DMSO-d6) δ 1.18 - 1.40 (m, 10H), 1.38 - 1.55 (m, 12H), 2.04 - 2.23 (m, 6H), 3. 29 - 3.38 (m, 2H), 3.40 - 3.51 (m, 2H), 4.39 (t, J = 5.6 Hz, 1H), 11.93 (s, 1H).

[0697] Step 3: 3-((6,6-bis((7,7,8,8,8-pentafluorooctyl )oxy)hexanoyl)oxy)-2-(hydroxymethyl)propyl(9Z,12Z )-Octadeca-9,12-dienoate

[0698] [ka]

[0699] 6,6-bis((7,7,8,8,8-pentafluorooctyl)oxy)hexanoic acid Prepared from according to general procedure C. Yield 140 mg (57%). 1 H NMR (400 MHz, Roform-d) δ 0.87 (d, J = 7.3 Hz, 3H), 1.22 - 1.46 (m, 27H), 1.56 - 1.70 (m, 9 H), 1.88 - 2.10 (m, 9H), 2.11 - 2.25 (m, 2H), 2.26 - 2.37 (m, 4H), 2.76 (t, J = 7.3 Hz, 2H), 3.36 - 3.44 (m, 2H), 3.49 - 3.58 (m, 2H), 3.60 (t, J = 5.7 Hz, 2H), 3.91 - 4.10 (m, 1H), 4.09 - 4.26 (m, 4H), 4.36 - 4.53 (m, 1H), 5.22 - 5.52 (m, 4H).

[0700] Step 4: 3-((6,6-bis((7,7,8,8,8-pentafluorooctyl )oxy)hexanoyl)oxy)-2-((((3-(diethylamino)propoxy) Carbonyl)oxy)methyl)propyl(9Z,12Z)-octadeca-9,12-dienes Nonoate (Example 122). 3-((6,6-bis((7,7,8,8,8-pentafluoro) (hexanoyl)oxy)-2-(hydroxymethyl)propyl( 9Z,12Z)-Octadeca-9,12-dienoate and 3-(diethylamino)- Prepared from 1-propanol according to general procedure D. Yield 51 mg (61%). LCMS (method Method B): (M+H) m / z = 1060.7, RT = 1.70 min.

[0701] [ka]

[0702] [Example 123] 3-((6,6-bis((7,7,8,8,8-pentafluorooctyl)oxy)hexyl ((((1-ethylpiperidin-3-yl)methoxy)-2-(((((1-ethylpiperidin-3-yl)oxy)-2-(((((1-ethylpiperidin-3-yl)methoxy ... (9Z,12Z)-octadeca-9,12-dienoic acid, (9Z,12Z)-(methyl)propyl 3-((6,6-bis((7,7,8,8,8-pentafluorooctyl)oxy) )hexanoyl)oxy)-2-(hydroxymethyl)propyl(9Z,12Z)-octanoyl Tadeca-9,12-dienoate and (1-ethylpiperidin-3-yl)methanol Prepared from according to general procedure D. Yield 80 mg (74%). LCMS (Method B): (M+H) Measured value m / z = 1073.2, RT = 1.81 minutes.

[0703] [ka]

[0704] [Example 124] 3-((4,4-bis(((Z)-oct-5-en-1-yl)oxy)butanoyl) oxy)-2-(((((1-(2-hydroxyethyl)piperidin-3-yl)meth (9Z,12Z)-octadeca-9,12-carbonyloxymethylpropyl Dienoate

[0705] Step 1: Ethyl 1-(2-hydroxyethyl)piperidine-3-carboxylate

[0706] [ka]

[0707] Ethyl piperidine-3-carboxylate (2 g, 12 mL) in anhydrous acetone (10 mL) To a stirred solution of 2-bromoethan-1-ol (1.89 g, 15.2 mmol) 8 mmol), dry powder K2CO3 (3.5 g, 25.47 mmol), and KI (0 The reaction mixture was stirred at 25°C for 18 hours. The reaction mixture was filtered and the filtrate was evaporated under reduced pressure. by Combi-Flash column chromatography eluting with 100% ethyl acetate-hexane Purification gave ethyl 1-(2-hydroxyethyl)piperidine-3-carboxylate (2 0.2g, 85%) as a colorless oil. 1 H NMR (400 MHz, chloroform-d) δ 1.17 - 1.29 (m, 3H), 1.46 - 1.62 (m, 2H), 1.60 - 1.76 (m, 1H), 1.81 - 1.95 (m, 1H), 2.14 - 2.19 (m, 1H), 2.38 (t, J = 10.4 Hz, 1H), 2.42 - 2.59 (m, 4H), 2.62 - 2.71 (m, 1H), 2.80 - 2.89 (m, 1H), 3.52 - 3.63 (m, 2H), 4.10 (q, J = 7.1 Hz, 2H).

[0708] Step 2: Ethyl 1-(2-((tert-butyldimethylsilyl)oxy)ethyl ) Piperidine-3-carboxylate

[0709] [ka]

[0710] Ethyl 1-(2-hydroxyethyl)piperidine-3-carboxylate in DCM (2 mL) To a stirred solution of silane (200 mg, 0.99 mmol), triethylamine (0.35 mL, 2.48 mmol) and tert-butyldimethylsilyl chloride (225 mg, 1 The reaction mixture was stirred at 25°C for 14 hours. The reaction mixture was quenched with water, extracted with DCM (50 mL), and washed with brine solution (25 mL). The organic layer was dried over anhydrous Na2SO4, filtered, and concentrated under reduced pressure. The crude material obtained was purified by CombiFlash chromatography eluting with 5% ethyl acetate-hexane. The resulting product was purified by chromatography to give ethyl 1-(2-((tert-butyldimethylsilyl) (Hydroxy)ethyl)piperidine-3-carboxylate (160 mg, 51%) was added to the adhesive Obtained as a physical object. 1 H NMR (400 MHz, chloroform-d) δ 0.04 (s, 6H), 0.87 (s, 9H), 1.23 (t, J = 7.1 Hz, 3H), 1.32 - 1.48 (m, 1H), 1.47 - 1.64 (m, 1H), 1.63 - 1.77 (m, 1H), 1.87 - 1.98 (m, 1H), 2.00 - 2.13 (m, 1H), 2.22 (t, J = 10.9 Hz, 1H), 2. 48 - 2.59 (m, 3H), 2.78 (d, J = 11.2 Hz, 1H), 3.04 (d, J = 13.4 Hz, 1H), 3.73 (t , J = 6.4 Hz, 2H), 4.11 (q, J = 7.1 Hz, 2H).

[0711] Step 3: (1-(2-((tert-butyldimethylsilyl)oxy)ethyl)pyrimidinyl) Peridine-3-yl)methanol

[0712] [ka]

[0713] Ethyl 1-(2-(tert-butyldimethylsilyl)oxy)- )ethyl)piperidine-3-carboxylate (500 mg, 1.58 mmol) and stirring The solution was treated with lithium aluminum hydride (1 M in THF) (3.2 mL, 3.17 mmol ) was added at 0° C. and the reaction mixture was stirred at 25° C. for 1 hour. Upon completion, the reaction mass was washed with saturated N The mixture was quenched with aqueous a2SO4 (10 mL) at 0 °C and filtered. The filtrate was concentrated under reduced pressure. and (1-(2-((tert-butyldimethylsilyl)oxy)ethyl)piperidine- 3-yl)methanol (400 mg, 92%) was obtained as a colorless oil. Carried on to next step without purification. 1 H NMR (400 MHz, chloroform-d) δ 0.04 (s, 6H), 0.87 (s, 9H), 1.07 - 1.22 (m, 1H), 1.49 - 1.62 (m, 1H), 1.62 - 1.72 (m , 1H), 1.72 - 1.85 (m, 3H), 2.11 (d, J = 10.5 Hz, 1H), 2.23 (d, J = 11.6 Hz, 1H) , 2.48 (t, J = 6.5 Hz, 2H), 2.55 - 2.72 (m, 1H), 2.82 (d, J = 10.8 Hz, 1H), 3.53 (dd, J = 5.8, 10.6 Hz, 1H), 3.57 - 3.70 (m, 1H), 3.73 (t, J = 6.4 Hz, 2H).

[0714] Step 4: 3-((4,4-bis(((Z)-oct-5-en-1-yl)oxy) )butanoyl)oxy)-2-(((((1-(2-((tert-butyldimethylsilyl (3-yl)oxy)ethyl)piperidin-3-yl)methoxy)carbonyl)oxy)methyl) Propyl (9Z,12Z)-octadeca-9,12-dienoate

[0715] [ka]

[0716] 3-((4,4-bis(((Z)-oct-5-en-1-yl)methyl)-2- ... )oxy)butanoyl)oxy)-2-(hydroxymethyl)propyl(9Z,12Z) -Octadeca-9,12-dienoate (Intermediate IVa) (100 mg, 0.14 mmol) To a stirred solution of 10 ml of pyridine (0.03 mL, 0.29 mmol), N,N-dimethylpyridinium Lysine-4-amine (5.3 mg, 0.04 mmol), and 4-nitrophenyl chloroformate Phenyl (72.36 mg, 0.29 mmol) was added and the mixture was stirred at 25°C for 1 hour. (1-(2-((tert-butyldimethylsilyl)oxy)ethyl)piperidine- 3-yl)methanol (158 mg, 0.58 mmol) and DIPEA (0.1 mL , 0.58 mmol) was added and stirred for a further 16 h. Dilute with 1 mL of HCl, wash with 1 M Na2CO3 solution (3 × 10 mL), and add water (10 mL). The combined organic portion was dried over anhydrous Na2SO4, filtered, and concentrated under reduced pressure. The crude material thus obtained was purified by column chromatography eluting with 2% MeOH-DCM. Purification by biflash column chromatography gave 3-((4,4-bis((( Z)-oct-5-en-1-yl)oxy)butanoyl)oxy)-2-(((((1 -(2-((tert-butyldimethylsilyl)oxy)ethyl)piperidin-3-yl )Methoxy)carbonyl)oxy)methyl)propyl(9Z,12Z)-octadeca-9 ,12-dienoate (50 mg, 35%) was obtained as a pale yellow oil. LCMS (Method B): Found m / z for (M+H) = 991.1, RT = 1.14 min.

[0717] Step 5: 3-((4,4-bis(((Z)-oct-5-en-1-yl)oxy) )butanoyl)oxy)-2-(((((1-(2-hydroxyethyl)piperidine-3 -yl)methoxy)carbonyl)oxy)methyl)propyl(9Z,12Z)-octadeca 3-(2 ... (4,4-bis(((Z)-oct-5-en-1-yl)oxy)butanoyl)oxy )-2-(((((1-(2-((tert-butyldimethylsilyl)oxy)ethyl) Piperidin-3-yl)methoxy)carbonyl)oxy)methyl)propyl(9Z,12 Z)-Octadeca-9,12-dienoate (60 mg, 0.061 mmol) was stirred in a solution The solution contained tetra-n-butylammonium fluoride (TBAF) (0.12 mL, 0.12 m mol) (1M solution in tetrahydrofuran) was added. The resulting solution was The resulting solution was diluted with DCM (5 mL) and stirred for 4 h. The organic layer was diluted with DCM (2×5 mL), brine (5 mL), and quenched with water (5 mL). The extract was dried over anhydrous Na2SO4, filtered, and concentrated under reduced pressure. The crude material was purified by CombiFlash column chromatography eluting with 2% MeOH-DCM. The product was purified by filtration to give 3-((4,4-bis(((Z)-oct-5-en-1-yl )oxy)butanoyl)oxy)-2-(((((1-(2-hydroxyethyl)piperidin Zin-3-yl)methoxy)carbonyl)oxy)methyl)propyl(9Z,12Z)- Octadeca-9,12-dienoate (35 mg, 66%) was obtained as a brown sticky gum. LCMS (Method B): (M+H) found m / z = 876.7, RT = 1.58 min.

[0718] [ka]

[0719] [Example 125] 3-((4,4-bis(((Z)-oct-5-en-1-yl)oxy)butanoyl) oxy)-2-(((((1-(3-hydroxypropyl)piperidin-3-yl)meth Oxy)carbonyl)oxy)methyl)propyl(9Z,12Z)-octadeca-9,12 -Dienoate

[0720] Step 1: Ethyl 1-(3-hydroxypropyl)piperidine-3-carboxylate to

[0721] [ka]

[0722] Ethyl piperidine-3-carboxylate (2 g, 12 mL) in anhydrous acetone (10 mL) To a stirred solution of 3-bromopropan-1-ol (2.12 g, 15.74 mmol) was added 3-bromopropan-1-ol (2.12 g, 15. 28 mmol), dry powdered K2CO3 (3.5 g, 25.47 mmol), and KI ( 0.42 g, 2.55 mmol) was added, and the reaction mixture was stirred at 25° C. for 16 hours. The reaction mixture was filtered and the filtrate was evaporated under reduced pressure. Combi-flash column chromatography eluting with 5% ethyl acetate-hexane was used. and purified to give ethyl 1-(3-hydroxypropyl)piperidine-3-carboxylate (2.0 g, 73%) was obtained as a colorless oil. 1 H NMR (400 MHz, chloroform-d) δ 1.23 (t, J = 7.1 Hz, 3H), 1.37 - 1.59 (m, 2H), 1.62 - 1.79 (m, 3H), 1.88 - 1.95 (m, 1H), 1.96 - 2.08 (m, 1H), 2.11 - 2.22 (m, 2H), 2.47 - 2.56 (m, 1H), 2.58 (t, J = 5.7 Hz, 2H), 2.84 (d, J = 11.3 Hz, 1H), 3.06 (d, J = 9.7 Hz, 1H), 3.76 (t, J = 5.2 Hz, 2H), 4.10 (q, J = 7.1 Hz, 2H).

[0723] Step 2: Ethyl 1-(3-((tert-butyldimethylsilyl)oxy)propyl (I)piperidine-3-carboxylate

[0724] [ka]

[0725] Ethyl 1-(3-hydroxypropyl)piperidine-3-carbohydrate in DCM (2 mL) To a stirred solution of 200 mg of hydroxylate (0.93 mmol), triethylamine (0.3 mL, 3.31 mmol) and tert-butyldimethylsilyl chloride (211 mg, 1 The reaction mixture was slowly warmed to 25°C and stirred for 14 hours. The reaction mixture was then quenched with water, extracted with DCM and washed with brine solution. The combined organic layers were dried over anhydrous Na2SO4, filtered, and concentrated under reduced pressure. The crude material obtained as above was purified by Combiflash chromatography eluting with 5% ethyl acetate-hexane. Purification by column chromatography gave ethyl 1-(3-((tert-butyldihydrochloride). Methylsilyl)oxy)propyl)piperidine-3-carboxylate (160 mg, 5 2%) as a sticky solid. 1 H NMR (400 MHz, chloroform-d) δ 0.03 (s, 6H) , 0.87 (s, 9H), 1.24 (t, J = 7.1 Hz, 3H), 1.34 - 1.63 (m, 2H), 1.63 - 1.75 (m, 3 H), 1.87 - 2.01 (m, 2H), 2.11 (t, J = 10.7 Hz, 1H), 2.35 - 2.44 (m, 2H), 2.47 - 2.59 (m, 1H), 2.75 (d, J = 11.2 Hz, 1H), 2.96 (d, J = 10.1 Hz, 1H), 3.63 (t, J = 6.3 Hz, 2H), 4.11 (q, J = 7.1 Hz, 2H).

[0726] Step 3: (1-(3-((tert-butyldimethylsilyl)oxy)propyl) Piperidin-3-yl)methanol

[0727] [ka]

[0728] 1-(3-((tert-butyldimethylsilyl)oxy)proline in THF (2 mL) A stirred solution of (p-( ... Lithium aluminum hydride (1 M in THF) (0.6 mL, 0.60 mmol) The addition was carried out at 0° C. and the reaction mass was stirred for 1 hour at 0° C. Upon completion, the reaction mass was added with saturated aqueous NaSO The mixture was quenched with a solution of 1-(3-((tert- Butyldimethylsilyl)oxy)propyl)piperidin-3-yl)methanol (80m g, 92%) as a colorless oil, which was used in the next step without further purification. Used. 1 H NMR (400 MHz, chloroform-d) δ 0.03 (s, 6H), 0.88 (s, 9H), 1.10 - 1 .23 (m, 1H), 1.49 - 1.87 (m, 7H), 1.95 - 2.24 (m, 2H), 2.32 - 2.41 (m, 2H), 2.50 - 2.67 (m, 1H), 2.77 (d, J = 11.0 Hz, 1H), 3.54 (dd, J = 5.7, 10.5 Hz, 1H), 3.5 9 - 3.71 (m, 3H).

[0729] Step 4: 3-((4,4-bis(((Z)-oct-5-en-1-yl)oxy) )butanoyl)oxy)-2-(((((1-(3-((tert-butyldimethylsilyl (N-1)oxy)propyl)piperidin-3-yl)methoxy)carbonyl)oxy)methyl ) Propyl (9Z,12Z)-octadeca-9,12-dienoate

[0730] [ka]

[0731] 3-((4,4-bis(((Z)-oct-5-ene-1- yl)oxy)butanoyl)oxy)-2-(hydroxymethyl)propyl(9Z,12 Z)-Octadeca-9,12-dienoate (Intermediate IVa) (50 mg, 0.07 mm To a stirred solution of 1,000 mg of pyridine (0.01 mL, 0.14 mmol), N,N-dimethylformamide (0.01 mL, 0.14 mmol), Pyridin-4-amine (2.65 mg, 0.02 mmol), and 4-nitrochloroformate 1. The mixture was stirred at room temperature for 1 hour. After 2 hours, (1-(3-((tert-butyldimethylsilyl)oxy)propyl)piperidine Zin-3-yl (83.06 mg, 0.28 mmol) and DIPEA (0.05 mL , 0.28 mmol) was added and stirred at 25° C. for 16 hours. Upon completion, the reaction mixture was Dilute with CM (10 mL), wash with 1 M sodium carbonate solution (3 × 10 mL), and add water (1 The combined organic portion was dried over Na2SO4 and filtered. The extract was concentrated under reduced pressure and subjected to CombiFlash column chromatography eluting with 2% MeOH-DCM. The product was purified by chromatography to give 3-((4,4-bis(((Z)-oct-5-ene-1 -yl)oxy)butanoyl)oxy)-2-(((((1-(3-((tert-butyl (dimethylsilyl)oxy)propyl)piperidin-3-yl)methoxy)carbonyl Oxy)methyl)propyl (9Z,12Z)-octadeca-9,12-dienoate (5 0 mg, 30%) as a pale yellow oil. LCMS (Method B): (M+H) found m / z = 1005. 1, RT = 1.12 minutes.

[0732] Step 5: 3-((4,4-bis(((Z)-oct-5-en-1-yl)oxy) )butanoyl)oxy)-2-(((((1-(3-hydroxypropyl)piperidine- 3-yl)methoxy)carbonyl)oxy)methyl)propyl(9Z,12Z)-octa Deca-9,12-dienoate (Example 125). In anhydrous THF (1.0 mL) at 0° C. 3-((4,4-bis(((Z)-oct-5-en-1-yl)oxy)butanoyl) oxy)-2-(((((1-(3-((tert-butyldimethylsilyl)oxy)prop propyl)piperidin-3-yl)methoxy)carbonyl)oxy)methyl)propyl(9 Z,12Z)-Octadeca-9,12-dienoate (85 mg, 0.085 mmol) To the solution, tetra-n-butylammonium fluoride (TBAF) (0.17 mL, 0.1 7 mmol) (1 M solution in tetrahydrofuran) was added. After warming to 25°C, the mixture was stirred for 4 hours. The resulting solution was diluted with DCM (10 mL) and The organic layer was quenched with water (5 mL). The extract was dried over anhydrous Na2SO4, filtered, and concentrated under reduced pressure. The crude material was purified by Combi-Flash column chromatography eluting with 1.5% MeOH-DCM. The product was purified by filtration to give 3-((4,4-bis(((Z)-oct-5-en-1-yl) ((((1-(3-hydroxypropyl)pyridyl)oxy)butanoyl)oxy)-2-( ... Peridine-3-yl)methoxy)carbonyl)oxy)methyl)propyl (9Z,12Z )-Octadeca-9,12-dienoate (36 mg, 52%) pale yellow sticky gum Obtained as pure. LCMS (Method B): (M+H) found m / z = 891.0, RT = 1.61 min.

[0733] [ka]

[0734] [Example 126] 3-((4,4-bis(((Z)-oct-5-en-1-yl)oxy)butanoyl) oxy)-2-((((3-((3-hydroxypropyl)(methyl)amino)propoxy (9Z,12Z)-octadeca-9,12-carbonyloxymethylpropyl Dienoate.

[0735] Step 1: 3,3'-(methylazanediyl)bis(propan-1-ol)

[0736] [ka]

[0737] 3-(methylamino)propan-1-ol (100 mg, 1 mL) in EtOH (2 mL) To a stirred solution of Na2CO3 (260 mg, 2.24 mmol) and 3-Chloropropanol (0.1 mL, 1.12 mmol) was added under an argon atmosphere. The reaction mixture was refluxed at 80° C. for 4 hours. Upon completion, the reaction mixture was concentrated under reduced pressure. The crude material thus obtained was purified by CombiFlash chromatography eluting with 2% acetone-hexane. The compound was purified by column chromatography to give 3,3'-(methylazanediyl)bis(methylazanediyl) (Propan-1-ol) (90 mg, 55%) was obtained as a colorless oil. 1 H NMR (400 MHz, DMSO-d6) δ 1.53 - 1.67 (m, 2H), 1.68 - 1.78 (m, 1H), 1.84 (t, J = 6.2 Hz, 1H), 2.26 (s, 2H), 2.52 (d, J = 6.7 Hz, 3H), 2.90 (t, J = 7.5 Hz, 1H), 3.39 - 3. 51 (m, 6H), 3.68 (t, J = 6.5 Hz, 1H).

[0738] Step 2: 3-((4,4-bis(((Z)-oct-5-en-1-yl)oxy) )butanoyl)oxy)-2-((((3-((3-hydroxypropyl)(methyl)a Amino)propoxy)carbonyl)oxy)methyl)propyl(9Z,12Z)-octadecyl Benzyl-9,12-dienoate (Example 126). Intermediate IVa and 3,3'-(methyl Prepared from (azanediyl)bis(propan-1-ol) according to general procedure D. Yield 1 8 mg (23%). LCMS (Method B): (M+H) found m / z = 864.7, RT = 1.55 min.

[0739] [ka]

[0740] [Example 127] 3-((4,4-bis(((Z)-oct-5-en-1-yl)oxy)butanoyl) oxy)-2-((((3-(diethylamino)propyl)carbamoyl)oxy)methyl (9Z,12Z)-propyl octadeca-9,12-dienoate: Intermediate IVa and and N1,N1-diethylpropane-1,3-diamine according to general procedure D Yield 32 mg (53%). LCMS (Method B): (M+H) found m / z = 848.0, RT = 1.55 min.

[0741] [ka]

[0742] [Example 128] 3-((4,4-bis(((Z)-oct-5-en-1-yl)oxy)butanoyl) oxy)-2-((((2-(1-ethylpyrrolidin-2-yl)ethoxy)carbonyl )Oxy)methyl)propyl (9Z,12Z)-octadeca-9,12-dienoate

[0743] Step 1: 2-(1-ethylpyrrolidin-2-yl)ethan-1-ol

[0744] [ka]

[0745] The following is representative of general procedure G. 2-(pyrrolidine)-2-pyrrolidine in acetonitrile (1 mL) To a stirred solution of (20 mg, 0.21 mmol) diethyl ether (2-azin-2-yl)ethan-1-ol, K CO3 (72 mg, 0.65 mmol) and EtI (0.01 mL, 0.21 mmol) l) was added under nitrogen atmosphere. The resulting mixture was stirred at 56°C for 16 hours. Cool to room temperature, filter through a bed of celite, and evaporate the filtrate under reduced pressure to give 2-(1-ethyl) (Iron-pyrrolidin-2-yl)ethan-1-ol (18 mg, 58%) was obtained as a colorless liquid. Ta. 1 H NMR (400 MHz, chloroform-d) δ 1.08 (t, J = 7.2 Hz, 3H), 1.37 - 1.48 (m , 1H), 1.65 - 1.96 (m, 5H), 1.92 - 2.05 (m, 1H), 2.04 - 2.18 (m, 2H), 2.72 - 2.8 3 (m, 1H), 2.87 - 3.04 (m, 1H), 3.10 - 3.20 (m, 1H), 3.62 - 3.72 (m, 1H), 3.89 - 4.05 (m, 1H).

[0746] 3-((4,4-bis(((Z)-oct-5-en-1-yl)oxy)butanoyl )oxy)-2-((((2-(1-ethylpyrrolidin-2-yl)ethoxy)carbonyl (9Z,12Z)-octadeca-9,12-dienoate (Example 128) Intermediate IVa and 2-(1-ethylpyrrolidin-2-yl)ethane Prepared from -1-ol according to general procedure D. Yield 28 mg (52%). LCMS (Method B) : (M+H) Found m / z = 860.8, RT = 1.55 min.

[0747] [ka]

[0748] [Example 129] 3-((4,4-bis(((Z)-oct-5-en-1-yl)oxy)butanoyl) oxy)-2-((((((1R,3s,5S)-8-ethyl-8-azabicyclo[3. 2.1]octan-3-yl)oxy)carbonyl)oxy)methyl)propyl (9Z, 12Z)-Octadeca-9,12-dienoate

[0749] Step 1: (1R,3r,5S)-8-ethyl-8-azabicyclo[3.2.1]o Cetane-3-ol

[0750] [ka]

[0751] (1R,3r,5S)-8-azabicyclo[3.2.1]octan-3-ol Prepared according to procedure G. Yield 180 mg, 73%.

[0752] Step 2: -((4,4-bis(((Z)-oct-5-en-1-yl)oxy) butanoyl)oxy)-2-(((((1R,3s,5S)-8-ethyl-8-azabicyclo[4.2.1.2]ox ... Cyclo[3.2.1]octan-3-yl)oxy)carbonyl)oxy)methyl)pro Pyr(9Z,12Z)-octadeca-9,12-dienoate (Example 129). Intermediate IVa and (1R,3r,5S)-8-ethyl-8-azabicyclo[3.2.1]octyl Prepared from tan-3-ol according to general procedure D. Yield 16 mg (51%). LCMS (method Method B): (M+H) measured m / z = 872.8, RT = 1.70 min.

[0753] [ka]

[0754] [Example 130] 3-((4,4-bis(((Z)-oct-5-en-1-yl)oxy)butanoyl) oxy)-2-((((((1R,3r,5S)-8-ethyl-8-azabicyclo[3. 2.1]octan-3-yl)oxy)carbonyl)oxy)methyl)propyl (9Z, 12Z)-Octadeca-9,12-dienoate

[0755] Step 1: (1R,3r,5S)-8-ethyl-8-azabicyclo[3.2.1]o Cetane-3-ol

[0756] [ka]

[0757] (1R,3r,5S)-8-azabicyclo[3.2.1]octan-3-ol Prepared according to procedure G. Yield 80 mg (92%).

[0758] Step 2: 3-((4,4-bis(((Z)-oct-5-en-1-yl)oxy) )butanoyl)oxy)-2-(((((1R,3r,5S)-8-ethyl-8-aza Bicyclo[3.2.1]octan-3-yl)oxy)carbonyl)oxy)methyl)propion Dopyl(9Z,12Z)-octadeca-9,12-dienoate (Example 130). Compound IVa and (1R,3r,5S)-8-ethyl-8-azabicyclo[3.2.1]o Prepared from octan-3-ol according to general procedure D. Yield 16 mg (51%). LCMS ( Method B): (M+H) Found m / z = 872.9, RT = 1.64 min.

[0759] [ka]

[0760] [Example 131] 3-((4,4-bis(((Z)-oct-5-en-1-yl)oxy)butanoyl) oxy)-2-((((9Z,12Z)-octadeca-9,12-dienoyl)oxy) Methyl)propyl 1'-ethyl-[1,4'-bipiperidine]-4-carboxylate: Intermediate IVa and 1'-ethyl-[1,4'-bipiperidine]-4-carboxylic acid dihydrochloride Prepared from the salt according to general procedure E. Yield 138 mg (52%). LCMS (Method A): (M+H) Found m / z = 913.7, RT = 3.21 min.

[0761] [ka]

[0762] [Example 132] 3-((4,4-bis((7,7,8,8,8-pentafluorooctyl)oxy)buta 9Z,12Z)-octadeca-9,12-dienoyl)oxy)-2-((((9Z,12Z)-octadeca-9,12-dienoyl) Oxy)methyl)propyl 1'-ethyl-[1,4'-bipiperidine]-4-carboxy Rate: Intermediate IVf and 1'-ethyl-[1,4'-bipiperidine]-4-carvone Prepared from the acid dihydrochloride according to general procedure E. Yield 195 mg (78%). LCMS (Method A) : (M+H) Found m / z = 1097.5, RT = 3.19 min.

[0763] [ka]

[0764] [Example 133] 3-((4,4-bis(((Z)-oct-5-en-1-yl)oxy)butanoyl) oxy)-2-((((2-(1-ethylpiperidin-2-yl)ethoxy)carbonyl )Oxy)methyl)propyl (9Z,12Z)-octadeca-9,12-dienoate: From intermediate IVa and 2-(1-ethylpiperidin-2-yl)ethan-1-ol Prepared according to procedure D. Yield 38 mg (40%). LCMS (Method A): Found m / z for (M+H) = 874.7, RT = 3.98 minutes.

[0765] [ka]

[0766] [Example 134] 3-((4,4-bis(((Z)-oct-5-en-1-yl)oxy)butanoyl) Oxy)-2-((2-(1-ethylpiperidin-4-yl)acetoxy)methyl)propanol Pyr(9Z,12Z)-octadeca-9,12-dienoate: Intermediate IVa and 2- Prepared according to general procedure E from (1-ethylpiperidin-4-yl)acetic acid. 66 mg ( 72%). LCMS (Method A): (M+H) found m / z = 844.7, RT = 3.95 min.

[0767] [ka]

[0768] [Example 135] 3-((4,4-bis(oct-2-yn-1-yloxy)butanoyl)oxy)-2 -(((((1-ethylpiperidin-3-yl)methoxy)carbonyl)oxy)methyl ) Propyl (9Z,12Z)-octadeca-9,12-dienoate: In general procedure A The (Z)-oct-5-en-1-ol was replaced with oct-2-yn-1-ol. Prepared using the same procedure as in Example 3. Yield of final step: 91 mg (92%). LC MS (Method A): (M+H) found m / z = 856.7, RT = 3.63 min.

[0769] [ka]

[0770] [Example 136] 3-((4,4-bis(oct-2-yn-1-yloxy)butanoyl)oxy)-2 -((((3-(diethylamino)propoxy)carbonyl)oxy)methyl)propyl (9Z,12Z)-Octadeca-9,12-dienoate: (Z)- in General Procedure A The same procedure as in Example 1 was carried out, except that oct-5-en-1-ol was replaced with oct-2-yn-1-ol. Prepared using the same procedure. Yield for final step: 80 mg (82%). LCMS (Method A) : (M+H) Found m / z = 844.7, RT = 3.66 min.

[0771] [ka]

[0772] [Example 137] 3-((4,4-bis(non-2-yn-1-yloxy)butanoyl)oxy)-2- (((((1-ethylpiperidin-3-yl)methoxy)carbonyl)oxy)methyl) Propyl (9Z,12Z)-octadeca-9,12-dienoate: in general procedure A Example 1, replacing (Z)-oct-5-en-1-ol with non-2-yn-1-ol Prepared using a procedure similar to that of 3. Yield for final step: 86 mg (88%). LCMS ( Method A): (M+H) found m / z = 884.7, RT = 3.73 min.

[0773] [ka]

[0774] [Example 138] 3-((4,4-bis(non-2-yn-1-yloxy)butanoyl)oxy)-2- ((((3-(diethylamino)propoxy)carbonyl)oxy)methyl)propyl( 9Z,12Z)-Octadeca-9,12-dienoate: (Z)-O in General Procedure A The same procedure as in Example 1 was repeated except that 5-butan-1-ol was replaced with nonan-2-yn-1-ol. Prepared using procedure. Yield for final step: 87 mg (90%). LCMS (Method A): (M +H) observed m / z = 872.7, RT = 3.69 min.

[0775] [ka]

[0776] [Example 139] 3-((4,4-bis(oct-7-yn-1-yloxy)butanoyl)oxy)-2 -(((((1-ethylpiperidin-3-yl)methoxy)carbonyl)oxy)methyl ) Propyl (9Z,12Z)-octadeca-9,12-dienoate: In general procedure A The (Z)-oct-5-en-1-ol was replaced with oct-7-yn-1-ol. Prepared using the same procedure as in Example 3. Yield for final step: 111 mg (84%). LCMS (Method A): (M+H) found m / z = 856.7, RT = 3.74 min.

[0777] [ka]

[0778] [Example 140] 3-((4,4-bis(oct-7-yn-1-yloxy)butanoyl)oxy)-2 -((((3-(diethylamino)propoxy)carbonyl)oxy)methyl)propyl (9Z,12Z)-Octadeca-9,12-dienoate: (Z)- in General Procedure A The same procedure as in Example 1 was carried out, except that oct-5-en-1-ol was replaced with oct-7-yn-1-ol. Prepared using the same procedure. Yield for final step: 108 mg (83%). LCMS (Method A): (M+H) Found m / z = 844.7, RT = 3.76 min.

[0779] [ka]

[0780] [Example 141] 3-((4,4-bis(dec-2-yn-1-yloxy)butanoyl)oxy)-2- (((((1-ethylpiperidin-3-yl)methoxy)carbonyl)oxy)methyl) Propyl (9Z,12Z)-octadeca-9,12-dienoate: in general procedure A In Example 1, (Z)-oct-5-en-1-ol was replaced with dec-2-yn-1-ol. Prepared using a similar procedure to 3. Yield for final step: 122 mg (88%). LCMS (Method A): (M+H) found m / z = 912.8, RT = 4.13 min.

[0781] [ka]

[0782] [Example 142] 3-((4,4-bis(dec-2-yn-1-yloxy)butanoyl)oxy)-2- ((((3-(diethylamino)propoxy)carbonyl)oxy)methyl)propyl( 9Z,12Z)-Octadeca-9,12-dienoate: (Z)-O in General Procedure A The same procedure as in Example 1 was repeated except that dec-5-en-1-ol was replaced with dec-2-yn-1-ol. Prepared using procedure. Yield for final step: 115 mg (84%). LCMS (Method A): Found m / z for (M+H) = 900.8, RT = 4.04 min.

[0783] [ka]

[0784] [Example 143] 3-((4,4-bis(dec-3-yn-1-yloxy)butanoyl)oxy)-2- (((((1-ethylpiperidin-3-yl)methoxy)carbonyl)oxy)methyl) Propyl (9Z,12Z)-octadeca-9,12-dienoate: in general procedure A In Example 1, (Z)-oct-5-en-1-ol was replaced with dec-3-yn-1-ol. Prepared using a similar procedure to 3. Yield for final step: 104 mg (89%). LCMS (Method A): (M+H) found m / z = 912.8, RT = 3.97 min.

[0785] [ka]

[0786] [Example 144] 3-((4,4-bis(dec-3-yn-1-yloxy)butanoyl)oxy)-2- ((((3-(diethylamino)propoxy)carbonyl)oxy)methyl)propyl( 9Z,12Z)-Octadeca-9,12-dienoate: (Z)-O in General Procedure A The same procedure as in Example 1 was repeated except that dec-5-en-1-ol was replaced with dec-3-yn-1-ol. Prepared using procedure. Yield for final step: 108 mg (94%). LCMS (Method A): Found m / z for (M+H) = 900.8, RT = 3.93 min.

[0787] [ka]

[0788] [Example 145] 3-((4,4-bis(((E)-oct-2-en-1-yl)oxy)butanoyl) oxy)-2-(((((1-ethylpiperidin-3-yl)methoxy)carbonyl)o (Oxy)methyl)propyl (9Z,12Z)-octadeca-9,12-dienoate: Basic In step A, (Z)-oct-5-en-1-ol was converted to (E)-oct-2-en-1 Prepared using the same procedure as in Example 3, but replacing hydroxypropyl methylcellulose with hydroxypropyl methylcellulose. Yield of final step: 7 4 mg (89%). LCMS (Method A): (M+H) found m / z = 860.7, RT = 3.87 min.

[0789] [ka]

[0790] [Example 146] 3-((4,4-bis(((E)-oct-2-en-1-yl)oxy)butanoyl) oxy)-2-((((3-(diethylamino)propoxy)carbonyl)oxy)methyl (9Z,12Z)-propyl octadeca-9,12-dienoate: General procedure A (Z)-oct-5-en-1-ol to (E)-oct-2-en-1-ol Instead, it was prepared using the same procedure as in Example 1. Yield of the final step: 68 mg (8 3%). LCMS (Method A): (M+H) found m / z = 848.8, RT = 3.89 min.

[0791] [ka]

[0792] [Example 147] 3-((4,4-bis(((E)-non-2-en-1-yl)oxy)butanoyl)o oxy)-2-(((((1-ethylpiperidin-3-yl)methoxy)carbonyl)oxy (Ci)methyl)propyl (9Z,12Z)-octadeca-9,12-dienoate: Basic procedure In sequence A, (Z)-oct-5-en-1-ol is converted to (E)-non-2-en-1-ol. Prepared using the same procedure as in Example 3, but replacing ethanol. Yield of final step: 68 ml g (81%). LCMS (Method A): (M+H) found m / z = 888.8, RT = 4.01 min.

[0793] [ka]

[0794] [Example 148] 3-((4,4-bis(((E)-non-2-en-1-yl)oxy)butanoyl)o oxy)-2-((((3-(diethylamino)propoxy)carbonyl)oxy)methyl ) Propyl (9Z,12Z)-octadeca-9,12-dienoate: In general procedure A (Z)-oct-5-en-1-ol was replaced with (E)-non-2-en-1-ol. Prepared using the same procedure as in Example 1. Yield of final step: 34 mg (41% LCMS (Method A): (M+H) found m / z = 877.9, RT = 4.07 min.

[0795] lipid nanoparticles F. LNP Formulation The lipid nanoparticle components are dissolved in 100% ethanol at the molar ratio of the lipid components indicated. Nucleic acid (NA) cargo was dissolved in 10 mM citrate, 100 mM NaCl, pH 4.0 This resulted in a concentration of NA cargo of approximately 0.22 mg / mL. In terms of morphology, the NA cargo has a mass to barcode ratio of 1:10 to 10:1 for functional NAs. Functional NAs (e.g., siRNA, antisense, expressed DNA, mRNA) mixed in a ratio of ) and reporter DNA barcodes (previously described by Sago, 2018 PNAS). become.

[0796] The LNPs are formulated at a total lipid to NA mass ratio of 11.7. Precision Nanosystems NanoAssessors were used according to the protocol of the original author. Use the semblr Spark or Benchtop Instrument The lipid and NA solution are formed by microfluidic mixing. During mixing, differential pressure flow is used. Maintain a 2:1 ratio of aqueous to organic solvent. After mixing, collect the LNPs and add PBS. Dilute the 20 kDa filter (approximately 1:1 v / v) in PBS and incubate at 4°C for 8 to 24 hours. Further buffer exchange is performed using dialysis against a dialysis tank. After this initial dialysis, the individual L For each NP formulation, characterize via DLS to measure size and polydispersity. The pKa of a subpopulation of LNPs is measured via a TNS assay. LNPs encompassing a range of polydispersity and densities were pooled and loaded onto a 100 kDa dialysis cassette. The LNPs were further dialyzed against PBS at 4°C for 1-4 hours. After the second dialysis, the LNPs were diluted to 0.22 μL. Filter sterilize using an M filter and store at 4°C for further use.

[0797] G. LNP Characterization DLS - LNP hydrodynamic diameter and percent polydispersity (PDI%) were measured using Dynamic light scattering (DLS) (DynaPro plate reader II, Wyatt) was used. The LNPs were diluted to the appropriate concentrations with 1x PBS and analyzed.

[0798] Concentration and encapsulation efficiency - The concentration of NA was determined using Quinol as per the manufacturer's instructions. bit microRNA Kit (for siRNA) or HS RNA Kit (for mRNA The encapsulation efficiency is determined by measuring the ratio of undissolved LNP to dissolved LNP. Therefore, it is decided.

[0799] pKa - 10mM HEPES (Sigma Aldrich), 10mM ME S (Sigma Aldrich), 10 mM sodium acetate (Sigma), and 1 Prepare a stock solution of 40 nM sodium chloride (Sigma Aldrich) and Adjust the pH to a range of 4 to 10 using hydrogen and sodium hydroxide. Using four replicates per well, add 140 μL of pH-adjusted buffer to a 96-well plate. This was followed by 2-(p-toluidino)-6-naphthalenesulfonic acid (60 μg / m Add 5 μL of LNP to each well. Incubate for 5 minutes under gentle shaking. After incubation, the excitation wavelength was 325 nm and the emission wavelength was 435 nm (BioTek Fluorescence is measured using a Synergy H4 Hybrid.

[0800] LNP Administration - All studies involved the use of male and female mice approximately 8-12 weeks of age. Each mouse was temporarily restrained and pooled LNPs were collected from up to five mice per experiment. Administer the drug intravenously via tail vein injection to animals. Use age-matched mice and up to three mice per experiment. Animals are also administered vehicle (1x PBS) via tail vein injection. 72 hours after administration, liver Tissues including spleen, bone marrow and blood are collected for analysis.

[0801] Flow - Liver tissue is mechanically digested and then enzymatically digested using a mixture of proteinases The spleen tissue was then digested and passed through a 70 μM filter to generate a single cell suspension. All tissues were mechanically digested to generate single-cell suspensions. Red blood cells are lysed and then analyzed by flow cytometry and fluorescence-activated cell sorting (F Staining was performed with fluorescently labeled antibodies for BD FAC. All antibodies were commercially available. All samples were flow-tested using SMelody (Becton Dickinson). The cells are acquired by cytometry and gated before sorting. The structure of the target cells is as follows: size → single cell → live cell → target cell. T cells are CD45+CD3 +, monocytes as CD45+CD11b+, and B cells as CD45+CD In the liver, endothelial cells are defined as CD31+ and Kupffer cells as CD45+. Hepatocytes are defined as CD11b+ and hepatocytes as CD31- / CD45-. In this case, gating the downregulation of target genes is performed, whereas mRNA studies are performed. Gating for upregulation of target genes in vehicle-dosed mice Tissue from each cell line with the correct phenotype is used to set the gates for sorting. Sort up to 20,000 cells for cell subsets and add to 1x PBS After sorting, pellet the cells via centrifugation and follow the manufacturer's protocol. Quick Extract DNA Extraction Solution (L DNA was extracted using a ELISA kit (Diflunigen). DNA was stored at -20°C.

[0802] Barcode sequencing - DNA (genomic and DNA barcode) is sequenced using QuickE Extract (Lucigen) and purified by Illumina as previously described. a Sequenced using MiniSeq (Sago et al. PNAS 2018, Sago et al. JACs 2018, Sago, Lokugamage et al. Nano Letters 2018), frequency of DNA fragments in FACS-isolated samples A barcode count is normalized to the frequency of injected input. These data are Plotted as "Normalized Fold Above Input" can be.

[0803] H. Confirm LNP formulation The lipid nanoparticle components are dissolved in 100% ethanol at the molar ratio of the lipid components indicated. Nucleic acid (NA) cargo was dissolved in 10 mM citrate, 100 mM NaCl, pH 4.0 This resulted in a concentration of NA cargo of approximately 0.22 mg / mL. In the form, the NA cargo is a functional NA (e.g., siRNA, antisense, expressed DNA, The LNPs are formulated at a total lipid to NA mass ratio of 11.7. LNPs were prepared using a Precision Nanosystem according to the manufacturer's protocol. ms NanoAssemblr Spark or Benchtop Instrum The differential pressure flow rate is calculated by microfluidic mixing of lipid and NA solutions using a pressure sensor. Maintain a 3:1 ratio of aqueous to organic solvent during mixing using LN Collect P, dilute with PBS (approximately 1:1 v / v), and incubate in PBS for 8–24 h at 4 °C. Further buffer exchange is performed using dialysis against a 20 kDa filter. After analysis, each individual LNP formulation was analyzed using DLP to measure size and polydispersity. Characterize via S and measure pKa of LNP subpopulations via TNS assay After dialysis, the LNPs are filter sterilized using a 0.22 micron sterile filter and further Store at 4°C for use.

[0804] LNP characterization DLS - LNP hydrodynamic diameter and percent polydispersity (PDI%) were measured using Dynamic light scattering (DLS) (DynaPro plate reader II, Wyatt) was used. The LNPs are diluted to the appropriate concentration with 1x PBS and analyzed.

[0805] Concentration and encapsulation efficiency - The concentration of NA was determined using Quinol as per the manufacturer's instructions. bit microRNA Kit (for siRNA) or HS RNA Kit (for mRNA The encapsulation efficiency is determined by measuring the ratio of undissolved LNP to dissolved LNP. Therefore, it is decided.

[0806] pKa - 10mM HEPES (Sigma Aldrich), 10mM ME S (Sigma Aldrich), 10 mM sodium acetate (Sigma), and 1 Prepare a stock solution of 40 nM sodium chloride (Sigma Aldrich) and Adjust the pH to a range of 4 to 10 using hydrogen and sodium hydroxide. Using four replicates per well, add 140 μL of pH-adjusted buffer to a 96-well plate. This was followed by 2-(p-toluidino)-6-naphthalenesulfonic acid (60 μg / m Add 5 μL of LNP to each well. Incubate for 5 minutes under gentle shaking. After incubation, the excitation wavelength was 325 nm and the emission wavelength was 435 nm (BioTek Fluorescence is measured using a Synergy H4 Hybrid.

[0807] LNP Administration - All studies involved the use of male and female mice approximately 8-12 weeks of age. Each mouse was temporarily restrained and pooled LNPs were collected from up to five mice per experiment. Administer the drug intravenously via tail vein injection to animals. Use age-matched mice and up to three mice per experiment. Animals are also administered vehicle (1x PBS) via tail vein injection. 72 hours after administration, liver Tissues including spleen, bone marrow and blood are collected for analysis.

[0808] hEPO Expression - Human EPO (hEPO) protein expression was measured 6 hours after administration. The mice were temporarily restrained and bled (via the tail vein). Blood was collected in heparin tubes. The plasma was then processed and stored at -80°C until ready for use. The ELISA kit (DuoSet; DY28) manufactured by R&D systems was used. hEPO protein was measured using a ELISA kit (6-05) according to the manufacturer's instructions. Various exemplary ionizable lipids were added to LNPs A1-A17 using the following molar ratios: Formulated: lipid to nucleic acid (human EPO mRNA) mass ratios of 11.7:1, 18:1, or 25:1, 45% ionizable lipid / 9% distearoyl phosphate Diacetylcholine (DSPC) / 44% cholesterol / 2% PEG lipid or 50% iodide Polyvinylalanine lipids / 9% distearoylphosphatidylcholine (DSPC) / 38% cholesterol Activity was determined by administering a nucleic acid dose of 0.15 mg / kg intravenously as described. The results were determined by measuring hEPO expression in plasma 6 hours after intravenous injection. P encapsulation efficiency, hydrodynamic diameter, polydispersity index (PDI), and dose of LNP formulations Representative data for plasma hEPO measurements after 2 h are provided in Table 1.

[0809] [Table 1]

[0810] While various aspects and embodiments have been disclosed herein, other aspects and embodiments may be used. Various aspects and embodiments disclosed herein will be apparent to those skilled in the art. The embodiments are for illustrative purposes only and are not intended to be limiting. The scope and spirit of the present invention is indicated by the following claims.

Claims

1. Compounds of formula (I): 【Chemistry 1】 (In the formula, R １ is C ９ ~C ２０ Alkyl or C having 1 to 3 units of unsaturation ９ ~C ２０ Arke Nil; X １ and X ２ are each independently absent, or —O—, NR ２ - and 【Chemistry 2】 where each R ２ are independently hydrogen or C １ ~C ６ is alkyl; each a is independently an integer between 1 and 6; X ３ and X ４ are each independently absent, or one or two C １ ~C ６ 4- to 8-membered heterocyclyl optionally substituted with alkyl groups, 1 or 2 C １ ~C ６ 5-6 membered heteroaryl optionally substituted with alkyl groups, 1 or 2 C's １ ~C ６ 5-6 membered aryl optionally substituted with alkyl groups, 1 or 2 C's １ ~C ６ 4- to 7-membered cycloalkyl optionally substituted with an alkyl group, —O— , and -NR ３ -, where each R ３ are independently hydrogen atoms or C １ ~C ６ alkyl, where X １ -X ２ -X ３ -X ４ is an oxygen-oxygen bond, oxygen - does not contain any nitrogen or nitrogen-nitrogen bonds; X ５ Ha-(CH ２ ) ｂ - where b is an integer between 0 and 6; X ６ is hydrogen, C １ ~C ６ Alkyl, 1 or 2 C １ ~C ６ Required for alkyl groups optionally substituted 5-6 membered heteroaryl, or -NR ４ R ５ where R ４ and R ５ are each independently hydrogen or C １ ~C ６ alkyl; or alternatively To, R ４ and R ５ together with the nitrogen to which they are attached, form one or two C １ ~C ６ forming a 4- to 7-membered heterocyclyl optionally substituted with an alkyl group, wherein the heterocyclyl may contain an additional heteroatom selected from oxygen, sulfur, and nitrogen. Include as needed; Each X ７ are independently hydrogen, hydroxyl, or —NR ６ R ７ where R ６ and R ７ are each independently hydrogen or C １ ~C ６ alkyl; or alternatively , R ６ and R ７ together with the nitrogen to which they are attached, form one or two C １ ~C ６ forming a 4- to 7-membered heterocyclyl optionally substituted with an alkyl group, wherein the heterocyclyl may contain an additional heteroatom selected from oxygen, sulfur, and nitrogen. Include as needed; X １ , X ２ , X ３ , X ４ , and X ５ At least one of is present; A １ and A ２ are each independently C ５ ~C １２ Haloalkyl, C ５ ~C １２ Al Kenil, C. ５ ~C １２ Alkynyl, (C ５ ~C １２ alkoxy)-(CH ２ ) ｎ２ -, 1 or two halo groups, C １ ~C ６ Alkyl group, C １ ~C ６ haloalkyl group, or C １ ~ C ６ optionally ring-substituted with alkoxy groups (C ５ ~C １０ aryl)-(CH ２ ) ｎ ３ - and 1 or 2 C １ ~C ６ Optionally ring-substituted with alkyl groups (C ３ ~ C ８ cycloalkyl)-(CH ２ ) ｎ４ - selected from the group consisting of; or alternatively, A １ and A ２ together with the atoms to which they are attached, form one or two C ４ ~C １０ forming a 5- to 6-membered cyclic acetal substituted with an alkyl group; n1, n2, and n3 are each independently an integer between 1 and 4; and n4 is an integer between 0 and 4.

2. R １ is a C having two units of unsaturation ９ ~C ２０ The compound of claim 1, wherein the compound is alkenyl. Compound.

3. R １ teeth, 【Transformation 3】 2. The compound of claim 1, wherein:

4. X １ The compound according to any one of claims 1 to 3, wherein is absent.

5. X １ The compound according to any one of claims 1 to 3, wherein is -O-.

6. X １ teeth, 【Chemistry 4】 The compound according to any one of claims 1 to 5,

7. X ７ The compound of claim 6 , wherein is hydrogen.

8. X ２ The compound of any one of claims 1 to 7, wherein is absent.

9. X ２ teeth, 【Transformation 5】 The compound according to any one of claims 1 to 7,

10. X ７ The compound of claim 9 , wherein is hydrogen.

11. X ７ 10. The compound of claim 9, wherein is hydroxyl and a is 1.

12. X ７ Ha-NR ６ R ７ and a is 1.

13. X ３ The compound of any one of claims 1 to 12, wherein is absent.

14. X ３ The compound according to any one of claims 1 to 12, wherein is -O-.

15. X ３ is -NR ３ The compound according to any one of claims 1 to 12, wherein

16. X ３ is one or two C １ ~C ６ 4- to 8-membered heterocyclic group optionally substituted with alkyl groups The compound of any one of claims 1 to 12, which is telocyclyl.

17. X ３ is one or two C １ ~C ６ 5-6 membered alkyl group optionally substituted The compound of any one of claims 1 to 12, which is a heteroaryl.

18. X ３ is one or two C １ ~C ６ 5-6 membered alkyl optionally substituted with alkyl groups The compound according to any one of claims 1 to 12, which is a reel.

19. X ３ is one or two C １ ~C ６ 4- to 7-membered cyclohexyl ester optionally substituted with alkyl groups The compound of any one of claims 1 to 12, which is a chloroalkyl.

20. X ４ The compound of any one of claims 1 to 19, wherein is absent.

21. X ４ is —O—. thing.

22. X ４ is -NR ３ - according to any one of claims 1 to 13 or 16 to 19, compound.

23. X ４ is one or two C １ ~C ６ 4- to 8-membered heterocyclic group optionally substituted with alkyl groups 20. The compound of any one of claims 1 to 19, which is telocyclyl.

24. X ４ is one or two C １ ~C ６ 5-6 membered alkyl group optionally substituted The compound of any one of claims 1 to 19, which is a heteroaryl.

25. X ４ is one or two C １ ~C ６ 5-6 membered alkyl optionally substituted with alkyl groups The compound according to any one of claims 1 to 19, which is a reel.

26. X ４ is one or two C １ ~C ６ 4- to 7-membered cyclohexyl ester optionally substituted with alkyl groups The compound of any one of claims 1 to 19, which is a chloroalkyl.

27. The compound of any one of claims 1 to 26, wherein b is 0.

28. 27. The compound of any one of claims 1 to 26, wherein b is an integer between 1 and 6.

29. X ６ The compound of any one of claims 1 to 28, wherein is hydrogen.

30. X ６ is one or two C １ ~C ６ 5-6 membered alkyl group optionally substituted The compound of any one of claims 1 to 28, which is a heteroaryl.

31. X ６ is NR ４ R ５ The compound according to any one of claims 1 to 28,

32. R ４ and R ５ together with the nitrogen to which they are attached, form one or two C １ ~ C ６ forming a 4- to 7-membered heterocyclyl optionally substituted with an alkyl group; 31. The compound according to claim 31.

33. A １ and A ２ are each independently C ５ ~C １２ haloalkyl 33. The compound according to any one of claims 32.

34. A １ and A ２ are each independently C ５ ~C １２ alkenyl, claims 1 to 3 2. The compound according to any one of claims 1 to 11.

35. A １ and A ２ are each independently C ５ ~C １２ alkynyl, claims 1 to 3 2. The compound according to any one of claims 1 to 11.

36. A １ and A ２ are each independently ５ ~C １２ alkoxy)-(CH ２ ) ｎ２ The compound according to any one of claims 1 to 32, wherein

37. A １ and A ２ each independently represents one or two halo groups, C １ ~C ６ Alkyl group , C １ ~C ６ haloalkyl group, or C １ ~C ６ optionally ring-substituted with alkoxy groups; (C ５ ~C １０ aryl)-(CH ２ ) ｎ３ - any one of claims 1 to 32 The compound described in

38. A １ and A ２ each independently represents one or two C １ ~C ６ Required for alkyl groups Ring-substituted (C ３ ~C ８ cycloalkyl)-(CH ２ ) ｎ４ - is claim 1 33. The compound according to any one of claims 1 to 32.

39. A １ and A ２ together with the oxygen atoms to which they are attached, form one or two C ４ ~C １０ 3. Forming a 5- to 6-membered cyclic acetal substituted with an alkyl group, according to claims 1 to 3.

2. The compound according to any one of claims 1 to 11.

40. Examples 1 to 148 【Chemistry 6-1】 【Chemistry 6-2】 【Transformation 6-3】 【Chemistry 6-4】 【Transformation 6-5】 【Transformation 6-6】 [Transformation 6-7] [Transformation 6-8] 【Transformation 6-9】 【Chemistry 6-10】 【Chemistry 6-11】 【Chemistry 6-12】 【Chemistry 6-13】 【Chemistry 6-14】 【Chemistry 6-15】 【Chemistry 6-16】 【Chemistry 6-17】 【Chemistry 6-18】 【Chemistry 6-19】 【Chemistry 6-20】 【Chemistry 6-21】 【Chemistry 6-22】 【Chemistry 6-23】 [Chemistry 6-24] [Chemistry 6-25] [Chemistry 6-26] [Chemistry 6-27] 2. The compound of claim 1, wherein:

41. An ionizable lipid comprising a compound according to any one of claims 1 to 40; phospholipids; Polyethylene glycol-lipid; Cholesterol; and as needed nucleic acid A lipid nanoparticle composition comprising:

42. The amount of ionizable lipids is present in the range of about 35 to 65 mole percent relative to the total moles. The lipid nanoparticle composition of claim 41.

43. The lipid nanoparticle composition of claim 41 or 42, wherein the phospholipid is DSPC.

44. 43. The lipid nanoparticle composition of claim 41 or 42, wherein the phospholipid is DMPC.

45. The nucleic acid may be an siRNA, an miRNA, an mRNA, an expressed DNA, an antisense oligonucleotide, or the like. Any of claims 41 to 44, which is a nucleotide or an immunostimulatory oligonucleotide. A lipid nanoparticle composition described in claim 1.

46. A method for delivering a nucleic acid to a subject in need thereof, comprising the steps of: A method comprising administering to the subject the lipid nanoparticle composition described in paragraph 1.

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