Novel lipids for delivery of nucleic acid segments

Abstract

A compound of formula (I), formula (III) or formula (IIIa) or a pharma- ceutically acceptable salt thereof (A, L, X 1 , X 2 , a, b, R 1 and R 2 Disclosed herein are lipid nanoparticles comprising a compound of Formula (I), Formula (III) or Formula (IIIa) or a pharma- ceutically acceptable salt thereof; a pharmaceutical composition comprising a plurality of lipid nanoparticles comprising a compound of Formula (I), Formula (III) or Formula (IIIa) or a pharma- ceutically acceptable salt thereof and a nucleic acid segment; and a method for delivering a nucleic acid segment comprising administering a plurality of lipid nanoparticles comprising a compound of Formula (I), Formula (III) or Formula (IIIa) or a pharma- ceutically acceptable salt thereof and a nucleic acid segment.

Description

[Technical field]

[0001] CROSS-REFERENCE TO RELATED APPLICATIONS This application claims the benefit of priority to U.S. Provisional Patent Application No. 63 / 264,263, filed November 18, 2021, and U.S. Provisional Patent Application No. 63 / 374,756, filed September 7, 2022. Each of the above-listed applications is incorporated herein by reference in its entirety for all purposes. [Background technology]

[0002] Nucleic acid segments such as oligonucleotides (e.g., RNA, e.g., messenger RNA [mRNA] and small interfering RNA [siRNA], antisense oligonucleotides [ASO], and DNA) have broad potential as new therapeutic treatments for various diseases and disorders. However, challenges remain in the administration of oligonucleotide therapeutics. A typical formulation involves encapsulating oligonucleotides in lipid nanoparticles (LNPs). LNP formulations usually contain (a) ionizable or cationic lipids or polymeric substances with tertiary or quaternary amines to encapsulate polyanionic mRNA; (b) zwitterionic lipids similar to lipids in cell membranes; (c) cholesterol to stabilize the lipid bilayer of the LNP; and (d) polyethylene glycol (PEG)-lipids to provide a hydration layer to the nanoparticles, improve colloidal stability, and reduce protein absorption. (See Non-Patent Document 1).

[0003] In 2018, the FDA approved the first RNA interference therapeutic, PATISIRAN, for the treatment of polyneuropathy in humans with hereditary transthyretin-mediated amyloidosis, which is delivered intravenously using LNPs incorporating an ionizable lipid (DLin-MC3-DMA, [MC3]). However, MC3 may not be suitable for all delivery systems, depending on the targeted organ, the intended delivery route, and the therapeutic window sought. Studies in two toxicology-related test species, rats and monkeys, reported dose-limiting toxicity associated with LNP formulations based on MC3, but not with the delivered cargo (see Non-Patent Document 2). Recently, lipid nanoparticle technology has also been successfully applied to create the first approved mRNA product for vaccination against the SARS-COV-2 virus (see, for example, Non-Patent Document 3). However, there remains a need to develop novel ionizable lipids for use in lipid nanoparticle formulations for the delivery of oligonucleotide therapeutics. [Prior art documents] [Non-patent literature]

[0004] [Non-Patent Document 1] Kowalski et al.,Molecular Therapy,27(4),(2019),710-728 [Non-Patent Document 2] Sedic et al.,Vet. Pathol.55(2),(2018),341-354 [Non-Patent Document 3] Shoenmaker et al,International Journal of Pharmaceutics,601,(2021),120586 Summary of the Invention [Means for solving the problem]

[0005] In some embodiments, the compound of formula (I): [ka] or a pharma- ceutically acceptable salt thereof, A is, [ka] and a and b are each independently 6, 7, or 8; c, d, f and g are each independently 1 or 2; e is 0, 1 or 2; R 1 and R 2 are each independently [ka] and; h is 0, 1, 2 or 3; R 3 and R 4 are each independently -(CH2) i CH3; i is 3, 4, 5, 6 or 7 is disclosed.

[0006] In some embodiments, the compound of formula (III): [ka] or a pharma- ceutically acceptable salt thereof, A is a 4- to 6-membered monocyclic oxacyclyl or a 6- to 10-membered bicyclic oxacyclyl; L is a covalent bond or C1-C3 alkylene; a and b are each independently 5, 6, 7, or 8; R 1 and R 2 are each independently C7-C 11 Straight chain alkyl or C9-C 19 is a branched alkyl; 1 and R 2 At least one of them is C7-C 11 (not straight chain alkyl) is disclosed.

[0007] In some embodiments, the compound of formula (IIIa): [ka] or a pharma- ceutically acceptable salt thereof, A is a 4- to 6-membered monocyclic oxacyclyl or a 6- to 10-membered bicyclic oxacyclyl; L is a covalent bond or C1-C3 alkylene; X 1 and X 2 are each independently [ka] and *R 1 Show the connection points for; a and b are each independently 4, 5, 6, 7, 8, or 9; provided that when one of a and b is 4 or 5, the other is 6, 7, 8, or 9; R 1 and R 2 are each independently C7-C 11 Straight chain alkyl, C7-C 19 Branched alkyl or C7-C 19 alkylene-cyclopropylene-alkyl; 1 and R 2 At least one of them is C7-C 11 Not linear alkyl or at least one is C7-C 19 (Not alkylene-cyclopropylene-alkyl) is disclosed.

[0008] In some embodiments, lipid nanoparticles comprising a compound of Formula (I), Formula (III), or Formula (IIIa), or a pharma- ceutically acceptable salt thereof, are disclosed.

[0009] In some embodiments, a pharmaceutical composition is disclosed that includes a plurality of lipid nanoparticles comprising a compound of Formula (I), Formula (III), or Formula (IIIa), or a pharma- ceutically acceptable salt thereof, and a nucleic acid segment.

[0010] In some embodiments, disclosed are methods of treating a disease or disorder in a subject, comprising administering to the subject a therapeutically effective amount of a pharmaceutical composition as described herein.

[0011] In some embodiments, a pharmaceutical composition as described herein for use in treating a disease or disorder is disclosed. [Brief description of the drawings]

[0012] [Figure 1] FIG. 1 illustrates the expression of eGFP in rat lung homogenates 24 hours after intratracheal administration of LNP formulations containing Compound 1 and MC3. [Diagram 2] FIG. 2 illustrates BALF neutrophil concentrations in rat BALF 24 hours after intratracheal administration of LNP formulations containing Compound 1 and MC3. [Diagram 3] FIG. 3 illustrates the expression of eGFP in rat hearts 24 hours after intracardiac administration of LNP formulations containing Compound 1 and MOD5. [Figure 4] FIG. 4 illustrates the ratio of eGFP expressed in rat liver / rat heart 24 hours after intracardiac administration of LNP formulations containing Compound 1 and MOD5. [Diagram 5] FIG. 5 illustrates plasma KC levels in rats 24 hours after intracardiac administration of LNP formulations containing Compound 1 and MOD5. [Figure 6] FIG. 6 illustrates plasma IL-6 levels in rats 24 hours after intracardiac administration of LNP formulations containing Compound 1 and MOD5. [Figure 7] FIG. 7 illustrates plasma IP-10 levels in rats 24 hours after intracardiac administration of LNP formulations containing Compound 1 and MOD5. [Figure 8] FIG. 8 illustrates plasma MCP-1 levels in rats 24 hours after intracardiac administration of LNP formulations containing Compound 1 and MOD5. [Figure 9]FIG. 9 illustrates the expression of eGFP in mouse liver 24 hours after intravenous administration of LNP formulations containing Compound 1, Compound 2, Compound 3, Compound 4, Compound 5, and MC3. [Figure 10] FIG. 10 illustrates the expression of eGFP in mouse muscle 24 hours after intramuscular administration of LNP formulations containing Compound 1, Compound 4, and MOD8. [Figure 11] FIG. 11 illustrates the expression of eGFP in mouse liver 24 hours after intramuscular administration of LNP formulations containing Compound 1, Compound 4, and MOD8. DETAILED DESCRIPTION OF THE PREFERRED EMBODIMENTS

[0013] In some embodiments, the compound of formula (I): [ka] or a pharma- ceutically acceptable salt thereof, A is, [ka] and; a and b are each independently 6, 7, or 8; c, d, f and g are each independently 1 or 2; e is 0, 1 or 2; R 1 and R 2 are each independently [ka] and; h is 0, 1, 2 or 3; R 3 and R 4 are each independently -(CH2) i CH3; i is 3, 4, 5, 6 or 7 is disclosed.

[0014] In some embodiments of the compounds of formula (I), A is, [ka] where: c, d, f and g are each independently 1 or 2; e is 0, 1 or 2.

[0015] In some embodiments of the compounds of Formula (I), e is 0 or 1.

[0016] In some embodiments of the compounds of Formula (I), e is 0.

[0017] In some embodiments of the compounds of Formula (I), e is 1.

[0018] In some embodiments of the compounds of formula (I), A is, [ka] is selected from.

[0019] In some embodiments of the compound of Formula (I), a and b are each independently 6, 7, or 8;

[0020] In some embodiments of the compounds of Formula (I), a is 7.

[0021] In some embodiments of the compounds of Formula (I), b is 7.

[0022] In some embodiments of the compounds of formula (I), R 1 and R 2 are each independently [ka] where: h is 1 or 2; R 3 and R 4 are each independently -(CH2) iCH3; i is 3, 4, 5, 6 or 7.

[0023] In some embodiments of the compounds of Formula (I), h is 2.

[0024] In some embodiments of the compounds of Formula (I), i is 3, 4 or 5.

[0025] In some embodiments of the compounds of Formula (I), i is 4.

[0026] In some embodiments of the compounds of Formula (I), R 3 is -(CH2)4CH3.

[0027] In some embodiments of the compounds of Formula (I), R 4 is -(CH2)4CH3.

[0028] In some embodiments of the compounds of Formula (I), R 3 and R 4 are -(CH2)4CH3, respectively.

[0029] In some embodiments, the compound of formula (III): [ka] or a pharma- ceutically acceptable salt thereof, A is a 4- to 6-membered monocyclic oxacyclyl or a 6- to 10-membered bicyclic oxacyclyl; L is a covalent bond or C1-C3 alkylene; a and b are each independently 5, 6, 7, or 8; R 1 and R 2 are each independently C7-C 11 Straight chain alkyl or C9-C 19 is a branched alkyl; 1 and R 2 At least one of them is C7-C 11 (not straight chain alkyl) is disclosed.

[0030] In one embodiment of the compound of formula (III), R 1 and R 2 together contain up to 31 carbon atoms. In another embodiment of the compound of formula (III), R 1 and R 2 together contain 22 or more carbon atoms. In another embodiment of the compound of formula (III), R 1 and R 2 Together they contain 24 to 30 carbon atoms.

[0031] In some embodiments, the compound of formula (IIIa): [ka] or a pharma- ceutically acceptable salt thereof; A is a 4- to 6-membered monocyclic oxacyclyl or a 6- to 10-membered bicyclic oxacyclyl; L is a covalent bond or C1-C3 alkylene; X 1 and X 2 are each independently [ka] and *R 1 Show the connection points for; a and b are each independently 4, 5, 6, 7, 8, or 9; provided that when one of a and b is 4 or 5, the other is 6, 7, 8, or 9; i.e., when a is 4 or 5, b is 6, 7, 8, or 9; when b is 4 or 5, a is 6, 7, 8, or 9; R 1 and R 2 are each independently C7-C 11 Straight chain alkyl, C7-C 19 Branched alkyl or C7-C 19 alkylene-cyclopropylene-alkyl; 1 and R 2 At least one of them is C7-C11 (b) R 1 and R 2 At least one of them is C7-C 19 In some embodiments, R is not an alkylene-cyclopropylene-alkyl. 1 and R 2 have the same number of carbon atoms, and R 1 and R 2 may have the same or different chemical structures. 1 and R 2 have different numbers of carbon atoms.

[0032] In one embodiment of the compound of Formula (IIIa), R 1 and R 2 together contain up to 31 carbon atoms. In another embodiment of the compound of formula (IIIa), R 1 and R 2 together contain 22 or more carbon atoms. In another embodiment of the compound of formula (IIIa), R 1 and R 2 Together they contain 24 to 30 carbon atoms.

[0033] In some embodiments of the compound of Formula (IIIa), X 1 and X 2 Both are [ka] It is.

[0034] In some embodiments of the compound of Formula (IIIa), X 1 teeth, [ka] and X 2 teeth, [ka] It is.

[0035] In some embodiments of the compound of Formula (III) or Formula (IIIa), A is, [ka] and; c is 0, 1 or 2; d is 1, 2, or 3; provided that the sum of c and d is 2 to 4; f is 0, 1 or 2; g is 1, 2, or 3; with the proviso that the sum of f and g is 2 to 4.

[0036] In some embodiments of the compound of Formula (III) or Formula (IIIa), L is a covalent bond, -CH2-, or -CH2CH2-.

[0037] In some embodiments of the compound of Formula (III), R 1 and R 2 Both are C9-C 19 In some embodiments, R 1 and R 2 are two identical C9-C 19 It is a branched alkyl group.

[0038] In some embodiments of the compound of Formula (III), R 1 is C7-C 11 Straight chain alkyl or C9-C 19 is a branched alkyl; R 2 is C9-C 19 is a branched alkyl; 1 and R 2 are not identical. In some embodiments, R 1 and R 2 do not contain the same number of carbon atoms.

[0039] In some embodiments of the compound of Formula (IIIa), R 1 is C7-C 11 R is a straight chain alkyl; 2 is C7-C 19 In some embodiments, R 1is C7-C 11 R is a straight chain alkyl; 2 is C 11 -C 19 In another embodiment, R 1 is C7-C 11 R is a straight chain alkyl; 2 is C 13 -C 19 It is a branched alkyl.

[0040] In some embodiments of the compound of Formula (IIIa), R 1 is C7-C 19 Branched alkyl or C7-C 19 alkylene-cyclopropylene-alkyl; R 2 is C7-C 19 is a branched alkyl; 1 and R 2 are not the same. That is, R 1 and R 2 Both are C7-C 19 When R is a branched alkyl 1 and R 2 is R 1 and R 2 may have the same number of carbon atoms, but do not have the same chemical structure. 1 is C7-C 15 Branched alkyl or C7-C 15 alkylene-cyclopropylene-alkyl; R 2 is C 13 -C 19 In some embodiments, R 1 is C7-C 13 is a branched alkyl; R 2 is C 13 -C 19 It is a branched alkyl.

[0041] In some embodiments of the compound of Formula (IIIa), a and b are the same and are both 6, 7, or 8.

[0042] In some embodiments of a compound of Formula (IIIa), a and b are not the same and are each independently 4 to 9, with the proviso that (a) when one of a and b is 4 or 5, then the other is 6, 7, 8, or 9; and (b) the sum of a and b is 12 to 16.

[0043] In some embodiments of the compound of formula (III), R 1 is -(CH2) m -CH3 or [ka] and R 2 teeth, [ka] and m is 7, 8 or 9; n and h are each independently 0, 1, 2, or 3; R 3a and R 4a are each independently -(CH2) p CH3; R 3b and R 4b are each independently -(CH2) q CH3; p and q are each independently 0, 1, 2, 3, 4, 5, 6, 7, 8, or 9; 3a and R 4a contain at least 9 carbon atoms together with the carbon atom to which they are attached, and R 3b and R 4b together with the carbon atom to which they are attached contain at least 9 carbon atoms.

[0044] In some embodiments of the compound of Formula (III), R 1 and R 2 together contain 19 or fewer carbon atoms.

[0045] In some embodiments of the compound of Formula (IIIa), R 1 is -(CH2) m -CH3, [ka] and R 2 teeth, [ka] and m is 6, 7, 8 or 9; v is 1, 2 or 3; t is 3, 4, 5, 6, 7 or 8; n and h are each independently 0, 1, 2, or 3; R 3a and R 4a are each independently -(CH2) p CH3; R 3b and R 4b are each independently -(CH2) q CH3; R 5a is hydrogen or methyl; p and q are each independently 0, 1, 2, 3, 4, 5, 6, 7, 8, or 9; 3a , R 4a and R 5a contain at least 7 carbon atoms together with the carbon atom to which they are attached, and R 3b and R 4b together with the carbon atom to which they are attached contain at least 9 carbon atoms.

[0046] In some embodiments of the compound of Formula (IIIa), R 3a is -(CH2) p CH3, where p is 0, 1, 2 or 3; R 4a is -(CH2) q CH3, where p is 4, 5, 6, 7, 8 or 9.

[0047] In some embodiments of the compound of Formula (IIIa), R3b and R 4b Both are -(CH2) q CH3, where q is 5, 6, 7 or 8.

[0048] In some embodiments of the compound of formula (III), the compound has formula (IV): [ka] or a pharma- ceutically acceptable salt thereof; A is, [ka] and a and b are each independently 5, 6, 7, or 8; R 1 is C7-C 11 Straight chain alkyl or C9-C 19 is a branched alkyl; R 2 is C9-C 19 It is a branched alkyl.

[0049] In some embodiments of the compound of Formula (IV), R 1 and R 2 are both C 10 -C 17 It is a branched alkyl and contains the same number of carbon atoms.

[0050] In some embodiments of the compound of Formula (IV), R 1 is C7-C 11 Straight chain alkyl or C9-C 13 is a branched alkyl; R 2 is C 13 -C 19 is a branched alkyl; 1 and R 2 At least one of them is C 13 It is not a branched alkyl.

[0051] In some embodiments of the compound of Formula (IV), a and b are both 5, 6, 7, or 8.

[0052] In some embodiments of the compound of Formula (IIIa), the compound has the formula (IVa): [ka] or a pharma- ceutically acceptable salt thereof; A is, [ka] and; a and b are each independently 5, 6, 7, or 8; R 1 is C7-C 11 Straight chain alkyl, C7-C 15 Branched alkyl or C7-C 15 alkylene-cyclopropylene-alkyl; R 2 is C 15 -C 19 It is a branched alkyl.

[0053] In some embodiments of Formula (IVa), a and b are the same and both are 5, 6, 7, or 8.

[0054] In some embodiments of the compound of formula (III), the compound has formula (V): [ka] or a pharma- ceutically acceptable salt thereof; A is, [ka] and a and b are each independently 5, 6, 7, or 8; R 1 is C7-C 11 Straight chain alkyl or C9-C 19 is a branched alkyl; R 2 is C9-C 19 It is a branched alkyl.

[0055] In some embodiments of the compound of Formula (V), R 1 and R 2 are both C 10 -C 17 It is a branched alkyl and contains the same number of carbon atoms.

[0056] In some embodiments of the compound of Formula (V), R 1 is C7-C 11 Straight chain alkyl or C9-C 13 is a branched alkyl; R 2 is C 13 -C 19 is a branched alkyl; 1 and R 2 At least one of them is C 13 It is not a branched alkyl.

[0057] In some embodiments of the compound of Formula (V), a and b are both 5, 6, 7, or 8.

[0058] In some embodiments of the compound of Formula (IIIa), the is of Formula (Va): [ka] or a pharma- ceutically acceptable salt thereof; A is, [ka] and a and b are each independently 5, 6, 7, or 8; R 1 is C7-C 15 Branched alkyl or C7-C 15 alkylene-cyclopropylene-alkyl; R 2 is C 15 -C 19 It is a branched alkyl.

[0059] In some embodiments of Formula (Va), a and b are the same and both are 5, 6, 7, or 8.

[0060] In some embodiments of the compound of formula (III), the compound has formula (VI): [ka] or a pharma- ceutically acceptable salt thereof; A is, [ka] is selected from L is a covalent bond, -CH- or -CHCH-; a and b are each independently 5, 6, 7, or 8; R 1 is C7-C 11 Straight chain alkyl or C9-C 19 is a branched alkyl; R 2 is C9-C 19 It is a branched alkyl.

[0061] In some embodiments of the compound of Formula (VI), R 1 and R 2 are both C 10 -C 17 It is a branched alkyl and contains the same number of carbon atoms.

[0062] In some embodiments of the compound of Formula (VI), R 1 is C7-C 11 Straight chain alkyl or C9-C 13 is a branched alkyl; R 2 is C 13 -C 19 is a branched alkyl; 1 and R 2 At least one of them is C 13 It is not a branched alkyl.

[0063] In some embodiments of the compound of Formula (VI), a and b are both 5, 6, 7, or 8.

[0064] In some embodiments of the compound of Formula (IIIa), the compound has the formula (VIa): [ka] or a pharma- ceutically acceptable salt thereof; A is, [ka] Selected from; a and b are each independently 5, 6, 7, or 8; R 1 is C7-C 15 Branched alkyl or C7-C 15 alkylene-cyclopropylene-alkyl; R 2 is C 15 -C 19 It is a branched alkyl.

[0065] In some embodiments of Formula (VIa), a and b are the same and are both 5, 6, 7, or 8.

[0066] In some embodiments of the compound of formula (III), the compound has formula (VII): [ka] or a pharma- ceutically acceptable salt thereof; A is, [ka] and a and b are each independently 5, 6, 7, or 8; R 1 is C7-C 11 Straight chain alkyl or C9-C 19 is a branched alkyl; R 2 is C9-C19 It is a branched alkyl.

[0067] In some embodiments of the compound of Formula (VII), R 1 and R 2 are both C 10 -C 17 It is a branched alkyl and contains the same number of carbon atoms.

[0068] In some embodiments of the compound of Formula (VII), R 1 is C7-C 11 Straight chain alkyl or C9-C 13 is a branched alkyl; R 2 is C 13 -C 19 is a branched alkyl; 1 and R 2 At least one of them is C 13 It is not a branched alkyl.

[0069] In some embodiments of the compound of Formula (VII), a and b are both 5, 6, 7, or 8.

[0070] In some embodiments of the compound of Formula (IIIa), the compound has the formula (VIIa): [ka] or a pharma- ceutically acceptable salt thereof; A is, [ka] and; a and b are each independently 4, 5, 6, 7, or 8; provided that when one of a and b is 4 or 5, the other is 6, 7, 8, or 9; i.e., when a is 4 or 5, b is 6, 7, 8, or 9; when b is 4 or 5, a is 6, 7, 8, or 9; X 1 and X 2 are each independently [ka] and; *R 1 Show the connection points for; R 1 is C7-C 11 Straight chain alkyl or C7-C 15 is a branched alkyl; R 2 is C 15 -C 19 It is a branched alkyl.

[0071] In some embodiments of compounds of Formula (VIIa), a and b are both 6, 7, or 8; or alternatively, a and b are each independently 4 to 9; with the proviso that (a) when one of a and b is 4 or 5, then the other is 6, 7, 8, or 9; and (b) the sum of a and b is 12 to 16.

[0072] In some embodiments of the compound of Formula (VIIa), X 1 and X 2 Both [ka] or alternatively, X 1 teeth, [ka] and X 2 teeth, [ka] It is.

[0073] In some embodiments of the compound of Formula (III), Formula (IV), Formula (V), Formula (VI), or Formula (VII), R 1 teeth, [ka] is selected from.

[0074] In some embodiments of the compound of Formula (IIIa), Formula (IVa), Formula (Va), Formula (VIa), or Formula (VIIa), R 1 teeth, [ka] is selected from.

[0075] In some embodiments of the compound of Formula (III), Formula (IV), Formula (V), Formula (VI), or Formula (VII), R 2 teeth, [ka] is selected from.

[0076] In some embodiments of the compound of Formula (IIIa), Formula (IVa), Formula (Va), Formula (VIa), or Formula (VIIa), R 2 teeth, [ka] is selected from.

[0077] In some embodiments, the compound of formula (I) is a compound of formula (II): [ka] or a pharma- ceutically acceptable salt thereof, wherein A is as defined for formula (I).

[0078] In some embodiments, the compound of formula (I) is bis(3-pentyloctyl) 9-((2-oxaspiro[3.3]heptan-6-yl)amino)heptadecanedioate, or a pharma- ceutically acceptable salt thereof.

[0079] In some embodiments, the compound of formula (I) is bis(3-pentyloctyl) 9-((tetrahydro-2H-pyran-4-yl)amino)heptadecanedioate, or a pharma- ceutically acceptable salt thereof.

[0080] In some embodiments, the compound of formula (I) is bis(3-pentyloctyl) 9-(((tetrahydrofuran-3-yl)methyl)amino)heptadecanedioate, or a pharma- ceutically acceptable salt thereof.

[0081] In some embodiments, the compound of formula (I) is bis(3-pentyloctyl) 9-(((tetrahydro-2H-pyran-4-yl)methyl)amino)heptadecanedioate, or a pharma- ceutically acceptable salt thereof.

[0082] In some embodiments, the compound of formula (I) is bis(3-pentyloctyl) 9-((oxetan-3-ylmethyl)amino)heptadecanedioate, or a pharma- ceutically acceptable salt thereof.

[0083] In some embodiments, the compound of formula (I) is bis(3-pentyloctyl) 9-((2-oxaspiro[3.3]heptan-6-yl)amino)heptadecanedioate.

[0084] In some embodiments, the compound of formula (I) is bis(3-pentyloctyl) 9-((tetrahydro-2H-pyran-4-yl)amino)heptadecanedioate.

[0085] In some embodiments, the compound of formula (I) is bis(3-pentyloctyl) 9-(((tetrahydrofuran-3-yl)methyl)amino)heptadecanedioate.

[0086] In some embodiments, the compound of formula (I) is bis(3-pentyloctyl) 9-(((tetrahydro-2H-pyran-4-yl)methyl)amino)heptadecanedioate.

[0087] In some embodiments, the compound of formula (I) is bis(3-pentyloctyl) 9-((oxetan-3-ylmethyl)amino)heptadecanedioate.

[0088] In some embodiments, the compound of formula (III) or formula (IIIa) is selected from representative compounds in the present disclosure, such as compounds 6-57 as described herein or a pharma- ceutically acceptable salt thereof.

[0089] The compounds of formula (I), (II), (III), (IIIa), (IV), (IVa), (V), (Va), (VI), (VIa), (VII) and (VIIa), including any subgenus or species thereof, may have different isomeric forms. The terms "optical isomer", "stereoisomer" or "diastereoisomer" refer to any of the various stereoisomeric configurations that may exist for a given compound of formula (I), (II), (III), (IIIa), (IV), (IVa), (V), (Va), (VI), (VIa), (VII) and (VIIa), including any subgenus or species thereof. Since substituents may be attached to chiral centers of carbon atoms, it is understood that the compounds of the present disclosure include enantiomers, diastereomers and racemates. The term "enantiomer" includes pairs of stereoisomers that are non-superimposable mirror images of each other. A 1:1 mixture of a pair of enantiomers is a racemic mixture. This term is used to indicate a racemic mixture where appropriate. The terms "diastereomer" or "diastereoisomer" include stereoisomers that have at least two asymmetric atoms but are not mirror images of each other. Absolute stereochemistry is specified according to the Cahn-Ingold-Prelog RS system. When compounds are pure enantiomers, the stereochemistry at each chiral center may be specified by either R or S. Resolved compounds whose absolute configuration is unknown may be designated (+) or (-) depending on the way (dextrorotatory or levorotatory) they rotate plane polarized light at the wavelength of the sodium D line. Certain compounds of formula (I), (II), (III), (IIIa), (IV), (IVa), (V), (Va), (VI), (VIa), (VII) and (VIIa), including any subgenera or species thereof, may contain one or more asymmetric centers or axes and thus give rise to enantiomers, diastereomers or other stereoisomeric forms that may be defined with respect to absolute stereochemistry, such as (R)- or (S)-. The present disclosure is intended to include all such possible isomers, including racemic mixtures, optically pure forms and intermediate mixtures.Optically active (R)- and (S)-isomers may be prepared using chiral synthons or chiral reagents, or resolved using conventional techniques well known in the art, such as chiral HPLC.

[0090] As used herein, the term "alkyl" refers to a monovalent saturated hydrocarbon radical having a specified number of carbon atoms. Alkyl includes both straight-chain and branched alkyls. As used herein, the term "alkylene" refers to a divalent saturated hydrocarbon radical having a specified number of carbon atoms. Alkylene includes both straight-chain and branched alkylenes. As used herein, the prefix Cx-y when used in terms such as "Cx-y alkyl" or "Cx-y alkylene" (where x and y are integers) indicates the numerical range of carbon atoms present in the group. For example, C7-C 11 Alkyl refers to an alkyl radical having 7 to 11 carbon atoms. As used herein, the term "cyclopropylene" refers to a divalent saturated hydrocarbon radical derived from cyclopropane, such as the following structure: [ka] refers to a chemical moiety having the formula:

[0091] As used herein, the term "oxacyclyl" refers to a heterocyclyl having one or more oxygen atoms in the ring. In one embodiment, oxacyclyl refers to a ring moiety formed by carbon, oxygen, and hydrogen atoms. Oxacyclyl includes both monocyclic and bicyclic oxacyclyls. Bicyclic oxacyclyls include spiro bicyclics, where the two rings share only a single carbon atom, i.e., a spiro atom; in fused or fused bicyclics, the two rings share two adjacent atoms; in bridged bicyclics, the two rings share three or more atoms, with a bridge containing at least one atom separating the two bridgehead atoms. In one embodiment, a bicyclic oxacyclyl is a spiro bicyclic oxacyclyl. As used herein, the prefix X-Y members, when used in terms such as "X-Y membered oxacyclyl" (X and Y are integers), indicates the range of the number of atoms (i.e., carbon atoms and heteroatoms) present in the rings that form the ring structure. For example, a 4- to 6-membered oxacyclyl refers to an oxacyclyl having from 4 to 6 ring atoms.

[0092] In some embodiments, compounds of formula (I), formula (III) or any subgenus or species thereof are disclosed. In some embodiments, pharma- ceutically acceptable salts of compounds of formula (I), formula (III) or any subgenus or species thereof are disclosed. The term "pharma- ceutically acceptable salts" includes acid addition salts that retain the biological effectiveness and properties of compounds of formula (I), formula (III) or any subgenus or species thereof, and typically are not biologically or otherwise undesirable. Pharmaceutically acceptable acid addition salts may be formed with inorganic and organic acids, such as, for example, acetate, aspartate, benzoate, besylate, bromide / hydrobromide, bicarbonate / carbonate, hydrogensulfate / sulfate, camphorsulfonate, chloride / hydrochloride, chlortheophyllonate, citrate, ethanedisulfonate, fumarate, gluceptate, gluconate, glucuronate, hippurate, hydroiodide / iodide, isethionate, lactate, laurate, tetrahydrofuran ... Examples of suitable salts include but are not limited to: acetobionate, lauryl sulfate, malate, maleate, malonate, mandelate, mesylate, methylsulfate, naphthoate, napsylate, nicotinate, nitrate, octadecanoate, oleate, oxalate, palmitate, palmoate, phosphate / hydrogen phosphate / dihydrogen phosphate, polygalacturonate, propionate, stearate, succinate, subsalicylate, sulfate / hydrogen sulfate, tartrate, tosylate and trifluoroacetate. Inorganic acids from which salts can be derived include, for example, hydrochloric acid, hydrobromic acid, sulfuric acid, nitric acid, phosphoric acid, and the like. Organic acids from which salts can be derived include, for example, acetic acid, propionic acid, glycolic acid, oxalic acid, maleic acid, malonic acid, succinic acid, fumaric acid, tartaric acid, citric acid, benzoic acid, mandelic acid, methanesulfonic acid, ethanesulfonic acid, toluenesulfonic acid, trifluoroacetic acid, sulfosalicylic acid, and the like. In some embodiments, compounds of formula (II) are disclosed. In some embodiments, compounds of formula (II) are disclosed as pharma- ceutically acceptable salts thereof.

[0093] In some embodiments, lipid nanoparticles (LNPs) are disclosed that comprise a compound of formula (I), formula (III), or any subgenus or species thereof, or a pharma- ceutically acceptable salt thereof. In some embodiments, lipid nanoparticles (LNPs) are disclosed that comprise a compound of formula (I), formula (III), or any subgenus or species thereof. The term "lipid nanoparticles" includes electron-dense nanostructured cores that are generated by microfluidic mixing of lipid-containing solutions in ethanol and aqueous solutions. The lipid nanoparticles disclosed herein can be constructed from any material used in conventional nanoparticle technology, such as ionizable lipids, neutral lipids, sterols, and polymer-bound lipids, with the proviso that the net charge of the nanoparticles is about 0.

[0094] In some embodiments, the compound of formula (I), formula (III) or any subgenus or species thereof is an ionizable lipid. Other non-limiting examples of ionizable lipids that may be combined with the compound of formula (I), formula (III) or any subgenus or species thereof in lipid nanoparticles include lipids that contain a positive charge on the acidic scale, e.g., in the physiological pH range, such as 1,2-dilinoleyloxy-3-dimethylaminopropane (DLin-DMA), dilinoleylmethyl-4-dimethylaminobutyrate (DLin-MC3-DMA (see, e.g., U.S. Pat. No. 8,158,601 ) and the like. see detailed description), 2-Dilinoleyl-4-dimethylaminoethyl-[1,3]-dioxolane (DLin-KC2-DMA), Merck-32 (see, e.g., WO 2012 / 018754), Acuitas-5 (see, e.g., WO 2015 / 199952), KL-10 (see, e.g., U.S. Patent Application Publication No. 2012 / 0295832), C12-200 (see, e.g., Love, KT et al., PNAS, 107:1864 (2009)). The ionizable lipid may be present in an amount ranging from about 5% to about 90%, for example, about 10% to about 80%, for example, about 25% to about 75%, for example, about 40% to about 60%, about 40% to about 50%, for example, about 45% or about 50%, in terms of molar percentage relative to the total lipid present in the lipid nanoparticle.

[0095] The term "neutral lipid" includes lipids that have a net charge of zero at physiological pH, e.g., lipids that exist in uncharged or neutral zwitterionic form at physiological pH, e.g., distearoylphosphatidylcholine (DSPC), dioleoylphosphatidylethanolamine (DOPE), dipalmitoylphosphatidylcholine (DPPC), dimyristoylphosphatidylcholine (DMPC), and the like, and combinations thereof. The neutral lipid may be present in an amount ranging from about 1% to about 50%, e.g., from about 5% to about 20%, e.g., from 7.5% to about 12.5%, e.g., about 10%, by mole percent, relative to the total lipid present in the lipid nanoparticle. In some embodiments, the neutral lipid is DSPC. In some embodiments, the neutral lipid is DOPE. In some embodiments, the neutral lipid is DPPC. In some embodiments, the neutral lipid is DMPC.

[0096] The term "sterol" includes cholesterol and the like. The sterol may be present in an amount ranging from about 10% to about 90%, such as about 20% to about 50%, such as about 35% to 45%, such as about 38.5%, by mole percent relative to the total lipid present in the lipid nanoparticle. In some embodiments, the sterol is cholesterol.

[0097] The term "polymer-bound lipid" includes lipids that include a lipid moiety and a polymer moiety, such as PEGylated lipids that include both a lipid moiety and a polyethylene glycol moiety. Non-limiting examples include dimyristoylphosphatidylethanolamine-poly(ethylene glycol) 2000 (DMPE-PEG2000), DPPE-PEG2000, DMG-PEG2000, DPG-PEG2000, PEG2000-c-DOMG, PEG2000-c-DOPG, and the like. The molecular weight of the poly(ethylene glycol) that may be used may range from about 500 to about 10,000 Da or from about 1,000 to about 5,000 Da. In some embodiments, the polymer-bound lipid is DMPE-PEG2000. In some embodiments, the polymer-bound lipid is DPPE-PEG2000. In some embodiments, the polymer-bound lipid is DMG-PEG2000. In some embodiments, the polymer-bound lipid is DPG-PEG2000. In some embodiments, the polymer-bound lipid is PEG2000-c-DOMG. In some embodiments, the polymer-bound lipid is PEG2000-c-DOPG. The polymer-bound lipid may be present in an amount ranging from about 0% to about 20%, such as about 0.5% to about 5%, such as about 1% to about 2%, for example about 1.5%, by mole percent, relative to the total lipid present in the lipid nanoparticle.

[0098] In at least one embodiment of the present disclosure, lipid nanoparticles can be prepared by combining multiple lipid components.For example, lipid nanoparticles can be prepared by combining the compound of formula (I), formula (III) or any of its subgenera or species or its pharmaceutically acceptable salt, sterol, neutral lipid and polymer-bound lipid in a molar ratio of 50:40-x:10:x with respect to the total lipid present.For example, lipid nanoparticles can be prepared by combining the compound of formula (I), formula (III) or any of its subgenera or species or its pharmaceutically acceptable salt, sterol, neutral lipid and polymer-bound lipid in a molar ratio of 50:37:10:3 (mol / mol), or for example, 50:38.5:10:1.5 (mol / mol), or for example, 50:39.5:10:0.5 (mol / mol), or 50:39.75:10:0.25 (mol / mol).

[0099] In another embodiment, lipid nanoparticles can be prepared using a compound of formula (I), formula (III) or any subgenera or species thereof or a pharma- ceutically acceptable salt thereof, a sterol (such as cholesterol), a neutral lipid (such as DSPC) and a polymer-bound lipid (such as DMPE-PEG2000) in a molar ratio of about 50:38.5:10:1.5 (mol / mol) to the total lipid present. Another non-limiting example is a lipid nanoparticle comprising a compound of formula (I), formula (III) or any subgenera or species thereof or a pharma- ceutically acceptable salt thereof, a sterol (such as cholesterol), a neutral lipid (such as DSPC) and a polymer-bound lipid (such as DMPE-PEG2000) in a molar ratio of about 47.7:36.8:12.5:3 (mol / mol) to the total lipid present. Another non-limiting example is a lipid nanoparticle comprising a compound of formula (I), formula (III) or any subgenera or species thereof or a pharma- ceutically acceptable salt thereof, a sterol (such as cholesterol), a neutral lipid (such as DSPC) and a polymer-bound lipid (such as DMPE-PEG2000) in a molar ratio of about 52.4:40.4:6.4:0.8 (mol l / mol) to the total lipid present. In another embodiment, a non-limiting example is a lipid nanoparticle comprising a compound of formula (I), formula (III) or any subgenera or species thereof or a pharma- ceutically acceptable salt thereof, a sterol (such as cholesterol), a neutral lipid (such as DSPC) and a polymer-bound lipid (such as DMPE-PEG2000) in a molar ratio of about 53.5:41.2:4.6:0.7 (mol l / mol) to the total lipid present. Another non-limiting example is a lipid nanoparticle comprising a compound of Formula (I), Formula (III) or any subgenus or species thereof or a pharma- ceutically acceptable salt thereof, a sterol (such as cholesterol), a neutral lipid (such as DSPC) and a polymer-bound lipid (such as DMPE-PEG2000) in a molar ratio of about 30:50:19:1 (mol / mol) to the total lipid present.

[0100] The selection of neutral lipids, sterols and / or polymer-bound lipids comprising the lipid nanoparticles and the relative molar ratios of such lipids to each other can be determined by the characteristics of the selected lipids, the nature of the intended target cells and the characteristics of the nucleic acid segment to be delivered.For example, in certain embodiments, the molar percentage of the compound of formula (I), formula (III) or any subgenus or species thereof or pharma-ceutically acceptable salts thereof in the lipid nanoparticles can be greater than about 10%, greater than about 20%, greater than about 30%, greater than about 40%, greater than about 50%, greater than about 60%, or greater than about 70% based on the total lipids present.The molar percentage of neutral lipids in the lipid nanoparticles can be greater than about 5%, greater than about 10%, greater than about 20%, greater than about 30%, or greater than about 40% based on the total lipids present.The molar percentage of sterols in the lipid nanoparticles can be greater than about 10%, greater than about 20%, greater than about 30%, or greater than about 40% based on the total lipids present. The molar percentage of polymer-bound lipid in the lipid nanoparticles can be greater than about 0.25%, such as greater than about 1%, greater than about 1.5%, greater than about 2%, greater than about 5%, or greater than about 10%, relative to the total lipid present.

[0101] According to the present disclosure, lipid nanoparticles can comprise a compound of formula (I), formula (III) or any subgenus or species thereof or a pharmaceutically acceptable salt thereof, a neutral lipid, a sterol and / or a polymer-bound lipid, in any desired useful orientation. For example, the core of the nanoparticle can comprise a compound of formula (I), formula (III) or any subgenus or species thereof or a pharmaceutically acceptable salt thereof, alone or in combination with one or more layers comprising another ionizable lipid, a sterol and a neutral lipid, and / or a polymer-bound lipid can then surround the core. For example, according to one embodiment, the core of the lipid nanoparticle may comprise a core comprising a compound of formula (I), formula (III) or any subgenus or species thereof or a pharma- ceutical acceptable salt thereof and a sterol (e.g., cholesterol) in any particular ratio, surrounded by a neutral lipid monolayer (e.g., DSPC) of any particular thickness, and further surrounded by an outer polymer-linked lipid monolayer of any particular thickness. In such an example, the nucleic acid segment may be incorporated into the core or any one of the subsequent layers, depending on the nature of the intended target cell and the characteristics of the nucleic acid segment to be delivered. The core and outer layer may further comprise other components typically incorporated into lipid nanoparticles known in the art. Furthermore, it will be understood by those skilled in the art that liposomes are delivery vehicles with a vesicular structure that differs from lipid nanoparticles as disclosed herein. Liposome vesicles are composed of lipid bilayers that form in the shape of a hollow sphere surrounding an aqueous phase. For example, liposomes contain a lamellar phase, while lipid nanoparticles have a non-lamellar structure.

[0102] Additionally, the mole percentages of the lipid nanoparticle components (e.g., the compound of formula (I), formula (III) or any subgenera or species thereof or pharma- ceutically acceptable salts thereof, neutral lipids, sterols and / or polymer-bound lipids) that comprise the lipid nanoparticles can be selected to provide a particular physical parameter of the lipid nanoparticle as a whole, such as the surface area of ​​one or more of the lipids. For example, the mole percentages of the compound of formula (I), formula (III) or any subgenera or species thereof or pharma-ceutically acceptable salts thereof, neutral lipids, sterols and / or polymer-bound lipids that comprise the lipid nanoparticles can be selected to provide a surface area per neutral lipid, e.g., DSPC. As a non-limiting example, the mole percentages of the compound of formula (I), formula (III) or any subgenera or species thereof or pharma-ceutically acceptable salts thereof, neutral lipids, sterols and / or polymer-bound lipids that comprise the lipid nanoparticles can be selected to provide a surface area per neutral lipid, e.g., DSPC, of ​​about 1.0 nm 2 ~about 2.0nm 2 , for example, about 1.2 nm 2 The surface area per DSPC can be determined to give

[0103] In accordance with the present disclosure, the lipid nanoparticles may further comprise a therapeutically effective amount of a nucleic acid segment that may be associated on the surface of the lipid nanoparticle and / or encapsulated within the lipid nanoparticle.

[0104] The term "nucleic acid segment" is understood to mean any one or more nucleic acid segments selected from antisense oligonucleotides, DNA, mRNA, siRNA, Cas9 guide RNA complexes, or combinations thereof. The nucleic acid segments herein may be wild-type or modified. In at least one embodiment, the lipid nanoparticles may contain multiple different nucleic acid segments. In yet another embodiment, the wild-type or modified nucleic acid segments encode a polypeptide of interest. Modified nucleic acid segments include nucleic acid segments that have chemical modifications to any part of the structure such that the nucleic acid segment is not natural. In some embodiments, the nucleic acid segment is RNA. In some embodiments, the nucleic acid segment is mRNA. In some embodiments, the nucleic acid segment is modified mRNA.

[0105] The term "therapeutically effective amount" as used herein refers to an amount of a nucleic acid segment that is sufficient to modulate protein expression in a target tissue and / or cell type. In some embodiments, a therapeutically effective amount of a nucleic acid segment is an amount sufficient to treat a disease or disorder associated with the protein expressed by the nucleic acid segment.

[0106] In at least one embodiment, the weight ratio of the total lipid phase to the nucleic acid segment ranges from about 40:1 to about 1:1, such as about 10:1. This corresponds to an approximate molar ratio of the compound of formula (I), formula (III) or any subgenus or species thereof or a pharmaceutically acceptable salt thereof to the nucleic acid monomer of about 3:1. In yet another example, the weight ratio of the total lipid phase to the nucleic acid segment ranges from about 30:1 to about 1:1, such as about 20:1, which corresponds to an approximate molar ratio of the compound of formula (I), formula (III) or any subgenus or species thereof or a pharmaceutically acceptable salt thereof to the nucleic acid monomer of about 6:1. However, the relative molar ratio of the lipid phase and / or lipid phase components to the nucleic acid monomer may be determined by the nature of the intended target cells and the characteristics of the nucleic acid segment, and therefore is not limited to the scope of the above-identified embodiments. In some embodiments, the molar ratio of the compound of formula (I), formula (III) or any subgenera or species thereof or a pharmaceutically acceptable salt thereof to the nucleic acid monomer is about 2.75:1 to 6:1. In some embodiments, the molar ratio of the compound of formula (I), formula (III) or any subgenera or species thereof or a pharmaceutically acceptable salt thereof to the nucleic acid monomer is about 2.75:1. In some embodiments, the molar ratio of the compound of formula (I), formula (III) or any subgenera or species thereof or a pharmaceutically acceptable salt thereof to the nucleic acid monomer is about 3:1. In some embodiments, the molar ratio of the compound of formula (I), formula (III) or any subgenera or species thereof or a pharmaceutically acceptable salt thereof to the nucleic acid monomer is about 5.5:1. In some embodiments, the approximate molar ratio of the compound of Formula (I), Formula (III), or any subgenus or species thereof, or a pharma- ceutically acceptable salt thereof to the amount of nucleic acid monomer is about 6:1.

[0107] In some embodiments, the lipid nanoparticles have a z-average diameter (particle size) of about 200 nm or less, e.g., about 100 nm or less, or, e.g., about 75 nm or less. <d> Z In at least one embodiment of the present disclosure, the lipid nanoparticles have a z-average particle size ranging from about 50 nm to about 100 nm, such as from about 60 nm to about 90 nm, from about 70 nm to about 80, such as about 75 nm.

[0108] In certain embodiments, the lipid nanoparticles have an encapsulation efficiency (%EE) of the nucleic acid segment of about 80% or more, such as greater than about 90%, such as in the range of about 95% to 100%. As used herein, the term "encapsulation efficiency" refers to the ratio of the nucleic acid segment encapsulated in the lipid nanoparticle to the total nucleic acid segment content in the lipid nanoparticle composition, as measured by dissolving the lipid nanoparticle with a detergent, such as Triton X-100.

[0109] The pharmaceutical composition of the present disclosure may further comprise at least one pharma- ceutically acceptable carrier.As used herein, the term "pharmaceutically acceptable carrier" includes compounds, materials, compositions and / or dosage forms that are suitable for use in contact with human and animal tissues, within the scope of sound medical judgment, without excessive toxicity, irritation, allergic reaction or other problems or complications, commensurate with a reasonable benefit / risk ratio.

[0110] The pharmaceutical composition may be in a form suitable for parenteral administration. The pharmaceutical composition may be in a form suitable for intratracheal instillation, bronchial instillation and / or inhalation. The pharmaceutical liquid composition may be nebulized by use of an inert gas. Nebulized suspension may be directly inhaled from the nebulizing device or the nebulizing device may be attached to a face mask tent, or intermittent positive pressure breathing device.

[0111] The amount of nucleic acid segment combined with one or more pharma- ceutically acceptable carriers to produce a single dosage form will need to vary depending on the subject being treated and the particular route of administration. For further information regarding routes of administration and dosing regimens, the reader is referred to Chapter 25.3 in Volume 5 of Comprehensive Medicinal Chemistry (Corwin Hansch; Chairman of Editorial Board), Pergamon Press 1990.

[0112] In one embodiment, the present disclosure provides a method for administering a pharmaceutical composition comprising a plurality of lipid nanoparticles comprising a compound of Formula (I), Formula (III) or any subgenus or species thereof, or a pharma- ceutically acceptable salt thereof, and a therapeutically effective amount of a nucleic acid segment in a subject in need thereof.

[0113] The term "subject" includes warm-blooded mammals, such as primates, cows, pigs, sheep, dogs, cats, rabbits, rats, and mice. In some embodiments, the subject is a primate, such as a human. In some embodiments, the subject is in need of treatment (e.g., the subject will benefit biologically or medically from the treatment).

[0114] The lipid nanoparticles disclosed herein can further serve as a platform for selective delivery of nucleic acid segments, such as antisense oligonucleotides, DNA, mRNA, siRNA, Cas9-guide RNA complexes, to target cells and tissues. Thus, in one embodiment, a method for delivering a nucleic acid segment to a cell comprises contacting the cell in vitro or in vivo with a pharmaceutical composition comprising a plurality of lipid nanoparticles comprising a compound of formula (I), formula (III) or any subgenus or species thereof, or a pharma-ceutically acceptable salt thereof, and a therapeutically effective amount of a nucleic acid segment. In some embodiments, the nucleic acid segment modulates expression, for example, by enhancing or reducing expression, or by upregulating or downregulating expression of a polypeptide.

[0115] Another embodiment provides a method for delivering a therapeutically effective amount of a nucleic acid segment to a subject in need thereof, comprising administering to the subject a pharmaceutical composition comprising a plurality of lipid nanoparticles comprising a compound of Formula (I), Formula (III) or any subgenera or species thereof, or a pharma-ceutically acceptable salt thereof, and a therapeutically effective amount of the nucleic acid segment.

[0116] A pharmaceutical composition comprising a plurality of lipid nanoparticles comprising a compound of formula (I), formula (III) or any subgenera or species thereof or a pharmaceutically acceptable salt thereof and a nucleic acid segment as disclosed herein can be used to treat a wide variety of disorders and diseases characterized by underexpression of a polypeptide in a subject, overexpression of a polypeptide in a subject, and / or absence / presence of a polypeptide in a subject. Thus, a method of treating a subject suffering from a disease or disorder is disclosed, comprising administering to the subject a pharmaceutical composition comprising a plurality of lipid nanoparticles comprising a compound of formula (I), formula (III) or any subgenera or species thereof or a pharmaceutically acceptable salt thereof and a therapeutically effective amount of a nucleic acid segment.

[0117] Further disclosed is the use of a pharmaceutical composition comprising a plurality of lipid nanoparticles comprising a compound of Formula (I), Formula (III) or any subgenus or species thereof, or a pharma- ceutically acceptable salt thereof, and a therapeutically effective amount of a nucleic acid segment, for treating a disease or disorder.

[0118] Further disclosed is a pharmaceutical composition comprising a plurality of lipid nanoparticles comprising a compound of Formula (I), Formula (III) or any subgenera or species thereof, or a pharma- ceutically acceptable salt thereof, and a therapeutically effective amount of a nucleic acid segment, for use in treating a disease or disorder.

[0119] Further disclosed is a method for increasing protein expression in a cell, comprising administering a pharmaceutical composition comprising a plurality of lipid nanoparticles comprising a compound of formula (I), formula (III) or any subgenus or species thereof or a pharma-ceutically acceptable salt thereof and a nucleic acid segment to a subject in need of increasing protein expression in a cell. In at least one embodiment, protein expression can be increased for about 2 hours to up to 24 hours. In another embodiment, protein expression can be increased for about 3 hours to up to 72 hours. EXAMPLES

[0120] General method 1 H NMR: 300 MHz; Probe: 5 mm broadband liquid probe BBFO with ATM+Z PABBO BB-1H / D; Magnet: ULTRASHIELD™ 300; Crate: AVANCE III 300; Autosampler: SampleXpress™ 60; Software: Topspin 3. 400 MHz; Probe: 5 mm broadband liquid probe BBFO with ATM+Z PABBO BB-1H / D; Magnet ASCEND™ 400; Crate AVANCE III 300; Autosampler SampleXpress™ 60; Software: Topspin 3. All spectra were calibrated using TMS as an internal standard. 500 MHz; Probe: 5 mm Bruker Smart probe with ATM+Z PABBO 500S1-BBF-HD; Magnet: ASCEND™ 500; Console: AVANCE Neo 500; Autosampler: SampleXpress™ 60; Software: Topspin 4. Proton chemical shifts are expressed in parts per million (ppm, δ scale) and are stated relative to residual protium in the NMR solvent (chloroform-d: δ 7.26, methanol-d4: δ 3.31, DMSO-d6: δ 2.50). Data are expressed as: chemical shift, multiplicity (s=singlet, d=doublet, t=triplet, q=quartet, dd=doublet of doublets, dt=doublet of triplets, m=multiplet, br=broad, app=apparent), integration and coupling constant (J) in Hertz (Hz).

[0121] LCMS: Instrument Shimadzu LCMS-2020 coupled with DAD detector, ELSD detector and 2020EV MS; Column Shim-pack XR-ODS C18 (50x3.0mm, 2.2μm); Eluent A water (0.05% TFA), Eluent B MeCN (0.05% TFA); Gradient 5 to 95% B in 2.00 min, hold 0.70 min (Method A) or 60 to 95% B in 1.00 min, hold 1.70 min (Method B); Flow rate 1.20mL / min; PDA detection (SPD-M20A) 190-400nm. Mass spectrometer in ESI mode.

[0122] Method C: UPLC was performed using a Waters Acquity UPLC and Waters SQD mass spectrometer (column temperature 30° C., UV detection=210-400 nm, mass analysis=ESI with positive / negative switching) at a flow rate of 1 mL / min using a solvent gradient of 2 to 98% B over 1.5 min (total run time for equilibration back to starting conditions: 2 min), with A=0.1% formic acid in water, B=0.1% formic acid in acetonitrile (for acid treatment) or A=0.1% ammonium hydroxide in water, B=acetonitrile (for base function). For acidic analysis the column used was a Waters Acquity HSS T3, 1.8 mm, 2.1x30 mm, for base analysis the column used was a Waters Acquity BEH C18, 1.7 mm, 2.1x30 mm.

[0123] HPLC: Instrument Shimadzu LCMS-2020 coupled with DAD detector, CAD detector; Column Ascentis Express C18 (100x4.6mm), 2.7μm; Mobile phase A water (0.05% TFA), Mobile phase B MeCN; Gradient 10 to 95% B in 4 min, hold 8 min or as indicated, flow rate 1.50mL / min; Purity as area %.

[0124] Prep HPLC: Equipment Waters 2545 Binary Gradient Module, Waters 2767 Sample Manager, Waters 2489 UV / Visible Detector, Waters SQ Detector2. Method A: Column XSelect CSH Prep C18 OBD column, 19x250mm, 5μm; Mobile phase A water (0.05% TFA), Mobile phase B MeCN; Flow rate 25mL / min, gradient as indicated. Method B: Column SunFire C18 OBD, 19x250mm, 5μm; Mobile phase A water (0.05% TFA), Mobile phase B MeCN; Flow rate 60mL / min, gradient as indicated.

[0125] Abbreviation 1,2-DCE 1,2-dichloroethane DCM Dichloromethane DMSO Dimethyl sulfoxide DIEA N,N-Diisopropylethylamine DMAP N,N-Dimethylaminopyridine <d> N Number average particle size <d> Z z-average particle size EDCI 1-Ethyl-3-(3-dimethylaminopropyl)carbodiimide EE% Encapsulation Efficiency HPLC High Performance Liquid Chromatography NMP N-Methyl-2-pyrrolidone PDI polydispersity index PE Petroleum Ether (30-50) PBS Phosphate Buffered Saline rt room temperature THF Tetrahydrofuran Z-pot Zeta potential

[0126] Scheme 1 below illustrates the synthetic procedures for preparing Examples 1-5.

[0127] Scheme 1: [ka] reagent. a) NaOEt, EtOH; b) c. HCl, EtOAc; c) EDCI, NEt3 or DIEA, DCM; d) TCBC, DMAP, THF; e) NaBH(OAc)3, NaOAc, THF; f) NaBH4, AcOH, THF; g) TsCl, NEt3, DMAP, DCM.

[0128] Example 1. Synthesis of Compound 1 Intermediate 1: Bis(3-pentyloctyl) 9-oxoheptadecanedioate [ka] Step 1: Tetraethyl 8-oxopentadecane-1,7,9,15-tetracarboxylate Sodium ethanolate (16.8 g, 247 mmol) was added in one portion to diethyl 3-oxopentanedioate (25 g, 124 mmol) dissolved in ethanol (200 mL) at rt under nitrogen. The resulting mixture was stirred at 80° C. for 1 h, followed by the addition of ethyl 7-bromoheptanoate (103 g, 432.73 mmol). The reaction mixture was heated at reflux overnight, then concentrated, diluted with EtOAc (200 mL), washed twice with water (200 mL), dried over sodium sulfate, filtered, and evaporated to give the crude product. Purification by flash chromatography on silica gel (eluted with 0 to 20% EtOAc in PE). Pure fractions were evaporated to dryness to give tetraethyl 8-oxopentadecane-1,7,9,15-tetracarboxylate (50.0 g, 79%) as an orange oil. 1 H NMR(300MHz,DMSO-d6)δ 4.08(m,8H),3.44 - 3.28(m,2H),2.26(m,4H),1.82 - 1.65(m,4H),1.61 - 1.41(m,8H),1.35 - 1.13(m,20H).

[0129] Step 2: 9-oxoheptadecanedioic acid Tetraethyl 8-oxopentadecane-1,7,9,15-tetracarboxylate (48 g, 93.3 mmol) was added in one portion to conc. HCl (36%, 400 mL) and HOAc (200 mL) under nitrogen at 25° C. The resulting mixture was stirred under reflux for 15 h. The reaction mixture was cooled to rt and poured into water. The precipitate was collected by filtration. Recrystallization from acetone gave 9-oxoheptadecanedioic acid (11.0 g, 38%) as a white solid. 1 H NMR(300MHz,DMSO-d6)δ 11.95(br.s,2H),2.38(t,J=7.3Hz,4H),2.18(t,J=7.3Hz,4H),1.56 - 1.37(m,8H),1.24(q,J=4.7Hz,12H).

[0130] Step 3: Bis(3-pentyloctyl) 9-oxoheptadecanedioate (Intermediate 1) To a solution of 9-oxoheptadecanedioic acid (19.2 g, 61.03 mmol), 3-pentyloctan-1-ol (see WO 2013 / 086354, p191 for procedure to prepare 3-pentyloctan-1-ol) (26.3 g, 131 mmol) and DIEA (32.0 mL, 183 mmol) in DCM (250 mL) was added EDCI (29.3 g, 153 mmol) and DMAP (7.46 g, 61.0 mmol). The mixture was stirred for 15 h and quenched with water (50 mL). After addition of EtOAc (750 mL), the mixture was washed twice each with 2M HCl (100 mL), water (100 mL) and brine (100 mL), dried over sodium sulfate and evaporated. The residue was purified by flash chromatography (eluent 0 to 5% EtOAc in PE) to give bis(3-pentyloctyl) 9-oxoheptadecandioate (Intermediate 1, (25.1 g, 61%) as a pale yellow oil. 1 H NMR(300MHz,CDCl3)δ 4.07(t,J=7.1Hz,4H),2.37(t,J=7.4Hz,4H),2.27(t,J=7.5Hz,4H),1.68 - 1.48(m,12H),1.45 - 1.15(m,46H),0.88(t,J=6.8Hz,12H).

[0131] Compound 1: Bis(3-pentyloctyl) 9-((2-oxaspiro[3.3]heptan-6-yl)amino)heptadecanedioate [ka] To 2-oxaspiro[3.3]heptan-6-amine hydrochloride (2.313 g, 15.46 mmol) and bis(3-pentyloctyl) 9-oxoheptadecanedioate (3.5 g, 5.15 mmol) in DCE (30 mL) and NMP (12.00 mL) was added sodium triacetoxyborohydride (3.28 g, 15.46 mmol). The resulting mixture was stirred at rt for 15 h.

[0132] The reaction mixture was concentrated, diluted with EtOAc (150 mL) and washed successively with water (3x25 mL) and brine (3x25 mL). The organic layer was dried over Na2SO4, filtered and evaporated to give 3 g of crude product containing solvent as an orange oil. The crude product was purified by flash chromatography on silica gel, elution gradient 0 to 20% MeOH in DCM to give 3 g of crude product as a yellow oil. The crude product was purified by preparative HPLC (A: water (10 mM NH4HCO3), B: CAN, 90 to 95% B in 7 min; flow rate: 25 mL / min) to give 1.025 g (26%) of the title compound (compound 1) as a pale yellow oil after lyophilization. LCMS m / z776.6[M+H] + , t R 2.060 minutes (method B). HPLC purity 96.4%t R 9.301 (B 30 to 95% at 8.00 min, held for 4 min). 1 H NMR (300 MHz, CD3OD, 23 °C) δ 4.74 (s, 2H), 4.60 (s, 2H), 4.13 (t, 4H), 3.09 - 3.26 (m, 1H), 2.42 - 2.63 (m, 3H), 2.33 (t, 4H), 1.98 (ddd, 2H), 1.56 - 1.7 (m, 8H), 1.33 (m, 54H), 0.88 - 0.99 (t, 12H). Number of expected H: 93; assigned H: 92.

[0133] Example 2. Synthesis of Compound 2: (Bis(3-pentyloctyl) 9-[(oxan-4-yl)amino]heptadecanedioate) [ka] Compound 2 was prepared according to the protocol described for compound 1, starting from tetrahydro-2H-pyran-4-amine (156 mg, 1.55 mmol) and bis(3-pentyloctyl) 9-oxoheptadecanedioate (350 mg, 0.52 mmol). Purification by flash chromatography on silica gel (0-10% MeOH in DCM) followed by preparative HPLC: XSelect CSH F-Phenyl OBD, 19*250 mm, 5 μm; A: water (0.05% TFA), B: ACN; flow rate: 25 mL / min; gradient: 56 to 95% B in 7 min. The crude product was isolated as a pale yellow oil (232 mg, 59%). LCMS m / z 764.8[M+H]+,t R 2.077 minutes (method A). HPLC purity 93.8%, t R 9.427 min (10-95%B in 8 min, hold for 4 min). 1 H NMR (300 MHz, MeOD) δ 4.12 (4H, t), 4.05 (2H, dd), 3.48 (3H, t), 2.34 (4H, t), 2.02 (2H, m), 1.54 - 1.78 (14H, m), 1.21 - 1.53 (51H, m), 0.88 - 0.99 (12H, m). Number of expected H: 93; assigned H: 92.

[0134] Compound 2 (232 mg crude) obtained as above was dissolved in EtOH and further purified by preparative SFC: Stationary phase: DCPak PBT, 250*20mm ID, 5μm; Mobile phase: A: CO2, B: 20mM NH3 in MeOH; Flow rate: 100g / min; Gradient 12% B (3.5 min), 12 to 35% B (1 min), 35% B (4 min); Pressure: 120 bar; Temperature: 40°C. Yield: 110 mg.

[0135] SFC-CAD purity 99%, t R 2.432 min. Stationary phase: DCPak PBT, 150*4.6mm ID, 5μm; Mobile phase: A:CO2 B:20mM NH3 in MeOH; Flow rate 3.5mL / min; Gradient 5 to 40%B (5min), 40%B (1min); Pressure: 120bar; Temperature: 40℃.

[0136] Example 3. Synthesis of Compound 3: (bis(3-pentyloctyl) 9-(((tetrahydrofuran-3-yl)methyl)amino)heptadecanedioate) [ka] Compound 3 was prepared using intermediate 1 following the protocol described for compound 1. Sodium triacetoxyhydroborate (32.9 mg, 0.16 mmol) was added in one portion to a stirred solution of bis(3-pentyloctyl)9-oxoheptadecanedioate (58.6 mg, 0.09 mmol), (tetrahydrofuran-3-yl)methanamine (14.25 μl, 0.13 mmol) and acetic acid (181 μl, 0.18 mmol) in 1,2-DCE (2 mL) and NMP (0.5 mL) under argon. The resulting solution was stirred at 25° C. for 18 h. The reaction mixture was diluted with DCM (15 mL), water (5 mL) and saturated Na2CO3 (5 mL). The layers were separated and the aqueous layer was extracted three times with DCM (15 mL). The combined organic layers were dried over MgSO4, filtered and evaporated to dryness to give the crude product. The resulting residue was purified by flash silica chromatography with an elution gradient of 10 to 50% MeOH in DCM (containing 1% NH4OH). The product fractions were concentrated to dryness under reduced pressure to give compound 3 (0.017 g, 26.1%) as a colorless oil. 1 H NMR(500MHz, methanol-d4)δ ppm 0.9(t,J=7.1Hz,12H)1.3-1.4(m,48H)1.4-1.5(m,6H)1.5-1.7(m,9H)2.0-2.1(m,1H)2.3-2.3(m,4H)2.3-2.4(m,1H)2.5(br C 48 H 93 NO5 m / z calculated value 763.705 Measured value 764.8 [M+H] + (LCMS-Method C).

[0137] Example 4. Synthesis of Compound 4: (bis(3-pentyloctyl) 9-(((tetrahydro-2H-pyran-4-yl)methyl)amino)heptadecanedioate) [ka] Compound 4 was prepared using intermediate 1 following the protocol described for compound 1. (After 10 min) sodium triacetoxyhydroborate (35.2 mg, 0.17 mmol) was added in one portion to a stirred solution of bis(3-pentyloctyl)9-oxoheptadecanedioate (62.7 mg, 0.09 mmol), (tetrahydro-2H-pyran-4-yl)methanamine (15.64 μl, 0.14 mmol) and acetic acid (194 μl, 0.19 mmol) in 1,2-DCE (2 mL) and NMP (0.5 mL) under argon. The resulting solution was stirred at 25° C. for 40 h. The reaction mixture was diluted with DCM (15 mL), water (5 mL) and saturated Na2CO3 (5 mL). The layers were separated and the aqueous layer was extracted three times with DCM (3×15 mL). The combined organic layers were dried over MgSO4, filtered and concentrated to dryness under reduced pressure to give the crude product. The resulting residue was purified by flash silica chromatography with an elution gradient of 10 to 45% MeOH in DCM (containing 1% NH4OH). The product fractions were concentrated to dryness under reduced pressure to give compound 4 (0.034 g, 47.7%) as a colorless oil. 1 H NMR(500MHz, methanol-d4)δ ppm 0.9(t,J=7.0Hz,12H)1.2-1.4(m,52H)1.4-1.5(m,6H)1.5-1.8(m,9H)2.3(t,J=7.3Hz ,4H)2.5-2.6(m,3H)3.4-3.5(m,2H)3.9(dd,J=11.0,4.0Hz,2H)4.1(t,J=6.7Hz,4H);C 49 H 95 NO5 m / z calculated value 777.721 Measured value 778.7 [M+H] + (LCMS-Method C).

[0138] Example 5. Synthesis of Compound 5: (Bis(3-pentyloctyl) 9-((oxetan-3-ylmethyl)amino)heptadecanedioate) [ka] Compound 5 was prepared using intermediate 1 following the protocol described for compound 1. To a stirred solution of bis(3-pentyloctyl) 9-oxoheptadecanedioate (69.6 mg, 0.10 mmol) and oxetan-3-ylmethanaminium chloride (38.0 mg, 0.31 mmol) in 1,2-DCE (2 mL) and NMP (0.5 mL) under argon was added sodium triacetoxyhydroborate (65.2 mg, 0.31 mmol) in one portion. The resulting solution was stirred at 25° C. for 50 h. The reaction mixture was diluted with DCM (15 mL), water (5 mL) and saturated Na2CO3 (5 mL). The layers were separated and the aqueous layer was extracted three times with DCM (15 mL). The combined organic layers were dried over MgSO4, filtered and concentrated to dryness under reduced pressure to give the crude product. The resulting residue was purified by flash silica chromatography, elution gradient 10 to 50% MeOH in DCM (1% NH4OH). The product fractions were concentrated to dryness under reduced pressure to give bis(3-pentyloctyl) 9-((oxetan-3-ylmethyl)amino)heptadecanedioate (0.048 g, 62.8%) as a colorless oil. 1 H NMR(500MHz, methanol-d4)δ ppm 0.9(t,J=7.1Hz,12H)1.3-1.4(m,48H)1.4-1.5(m,6H)1.6-1.7(m,8H)2.3(t,J=7.4Hz,4H)2.5(t,J=5. 9Hz,1H)2.9(d,J=7.5Hz,2H)3.1-3.1(m,1H)4.1(t,J=6.8Hz,4H)4.4(t,J=6.0Hz,2H)4.8-4.8(m,2H);C 47 H 91 NO5 m / z calculated value 749.690 Measured value 750.6 [M+H] + (LCMS-Method C).

[0139] Scheme 2 below illustrates the synthetic procedure for preparing Examples 6-9. In step d below, when the respective amine was utilized as the free base, AcOH was used as an additive in the reductive amination.

[0140] Scheme 2: [ka] Reagents: a) NaOEt, EtOH; b) conc. HCl, AcOH; c) EDC. HCl, DIPEA, DMAP, DCM; d) NaBH(OAc)3, AcOH, 1,2-DCE:NMP (4:1)

[0141] Example 6. Synthesis of Compound 6: Bis(3-pentyloctyl) 9-(((tetrahydro-2H-pyran-2-yl)methyl)amino)heptadecanedioate Step a): Sodium ethanolate (2.52 g, 37.09 mmol) was added portionwise to a stirred solution of diethyl 3-oxopentanedioate (4.49 mL, 24.73 mmol) in anhydrous (99.5%) ethanol (14 mL) at 25 °C under argon over 10 min. The resulting suspension was stirred at 81 °C for 1 h. Ethyl 7-bromoheptanoate (12.05 mL, 61.82 mmol) was added dropwise to the reaction mixture and the suspension was stirred at 81 °C for an additional 18 h. The reaction mixture was cooled to RT, concentrated to dryness under reduced pressure, redissolved in DCM (50 mL), extracted three times with water (50 mL) and washed with saturated aqueous NaCl (50 mL). The organic layer was dried over MgSO4, filtered and concentrated under reduced pressure to give the crude product. The resulting residue was purified by flash silica chromatography, elution gradient 10 to 50% EtOAc in hexanes. The product fractions were concentrated to dryness under reduced pressure to give tetraethyl 8-oxopentadecane-1,7,9,15-tetracarboxylate (8.10 g, 63.6%) as a pale yellow oil. 1 H NMR (500 MHz, chloroform-d) δ ppm 1.2-1.4 (m, 24H) 1.5-1.9 (m, 8H) 2.2-2.3 (m, 4H) 3.4-3.5 (m, 2H) 4.1-4.2 (m, 8H).

[0142] Step b): To a stirred solution of tetraethyl 8-oxopentadecane-1,7,9,15-tetracarboxylate (8.1 g, 15.74 mmol) in acetic acid (20 mL) at 25° C., hydrochloric acid (33.3 mL, 406.06 mmol) was added slowly. The resulting solution was stirred at 102° C. for 18 h with a reflux condenser and an outlet to remove excess HCl gas. The reaction was cooled to RT and the reaction mixture was poured onto ice water (50 mL) and allowed to stand for 30 min. The precipitate was collected by filtration, washed with cold water (3×20 mL) and dried under vacuum to give 9-oxoheptadecanedioic acid (crude) as a light yellow solid. The crude product was purified by crystallization from acetone to give 9-oxoheptadecanedioic acid (1.381 g, 27.9%) as a white solid. 1 C 17 H 30 O5 m / z calculated value 314.209 measured value 313.1 [MH] - (LCMS).

[0143] Step c): To a stirred solution of 9-oxoheptadecanedioic acid (419 mg, 1.33 mmol), 3-pentyloctan-1-ol (see WO 2013 / 086354, p191 for procedure to prepare 3-pentyloctan-1-ol) (641 mg, 3.20 mmol), N,N-dimethylpyridin-4-amine (32.6 mg, 0.27 mmol) and N-ethyl-N-isopropylpropan-2-amine (836 μl, 4.80 mmol) in DCM (20 mL) at 25° C. under argon was added 3-(((ethylimino)methylene)amino)-N,N-dimethylpropan-1-amine hydrochloride (690 mg, 3.60 mmol) in one portion. The resulting solution was stirred at 25° C. for 50 h. The reaction mixture was diluted with DCM (25 mL), water (10 mL) and saturated aqueous NH4Cl (10 mL). The layers were separated and the aqueous layer was extracted three times with DCM (25 mL). The combined organic layers were dried over MgSO4, filtered and concentrated to dryness under reduced pressure to give the crude product. The resulting residue was purified by flash silica chromatography, elution gradient 10 to 40% EtOAc in hexanes. The product fractions were concentrated to dryness under reduced pressure to give bis(3-pentyloctyl) 9-oxoheptadecandioate (0.820 g, 91%) as a pale yellow oil. 1 H NMR(500MHz,chloroform-d)δ ppm 0.9-0.9(m,12H)1.2-1.3(m,46H)1.5-1.7(m,12H)2.3(t,J=7.6Hz,4H)2.4(t,J=7.4Hz,4H)4.1(t,J=7.1Hz,4H).

[0144] Step d): [ka] To a stirred solution of (tetrahydro-2H-pyran-2-yl)methanaminium chloride (20.07 mg, 0.13 mmol) and bis(3-pentyloctyl)9-oxoheptadecanedioate (42.8 mg, 0.06 mmol) in 1,2-DCE (2 mL) and NMP (0.5 mL) at 25° C. under argon was added sodium triacetoxyhydroborate (32.1 mg, 0.15 mmol) in one portion. The resulting solution was stirred at 25° C. for 30 h. The reaction mixture was diluted with DCM (15 mL) and water (5 mL) along with saturated Na2CO3 (10 mL). The layers were separated and the aqueous layer was extracted with DCM (3×15 mL). The combined organic layers were dried over MgSO4, filtered and concentrated to dryness under reduced pressure to give the crude product. The resulting residue was purified by flash silica chromatography, elution gradient 0 to 30% 20% MeOH / DCM (with 1% NH4OH) in DCM. The product fractions were concentrated to dryness under reduced pressure to give compound 6 (17.40 mg, 35.5%) as a colorless oil. 1 H NMR (500MHz, methanol-d4) δ ppm 0.9(t,J=6.9Hz,12H)1.2-1.4(m,51H)1.4-1.5(m,6H)1.5-1.6(m,12H)1.9(br C 49 H 95 NO5 m / z calculated value 777.721 measured value 778.9 [M+H] + (LCMS).

[0145] Example 7. Synthesis of Compound 7: Bis(3-pentyloctyl) 9-((2-(tetrahydro-2H-pyran-4-yl)ethyl)amino)heptadecanedioate [ka] Compound 7 was prepared using tetraethyl 8-oxopentadecane-1,7,9,15-tetracarboxylate, 9-oxoheptadecanedioic acid and bis(3-pentyloctyl) 9-oxoheptadecanedioate according to the protocol described for compound 6, with the following additional steps: Sodium triacetoxyhydroborate (43.7 mg, 0.21 mmol) was added in one portion to a stirred solution of 2-(tetrahydro-2H-pyran-4-yl)ethan-1-amine (0.026 mL, 0.19 mmol), acetic acid (0.012 mL, 0.21 mmol) and bis(3-pentyloctyl) 9-oxoheptadecanedioate (46.7 mg, 0.07 mmol) in DCE (2 mL) and NMP (0.5 mL) at 25° C. under argon. The resulting solution was stirred at 25° C. for 30 h. The reaction mixture was diluted with DCM (15 mL) and water (5 mL) along with saturated Na2CO3 (10 mL). The layers were separated and the aqueous layer was extracted with DCM (3x15 mL). The combined organic layers were dried over MgSO4, filtered and concentrated to dryness under reduced pressure to give the crude product. The resulting residue was purified by flash silica chromatography, elution gradient 0 to 30% 20% MeOH / DCM (with 1% NH4OH) in DCM. The product fractions were concentrated to dryness under reduced pressure to give compound 7 (23.20 mg, 42.6%) as a colorless oil. 1 H NMR (400MHz, methanol-d4) δ ppm 0.9(t,J=7.0Hz,12H)1.3(br s,50H)1.4-1.5(m,8H)1.6(br d,J=6.6Hz,11H)2.3(s,4H)2.5-2.6(m,1H)2.6-2.7(m,2H)3.4-3.5(m,2H)3.9-4.0(m,2H)4.1-4.1(m,4H);C 50 H 97 NO5 m / z Calculated value 791.737 Measured value 792.8 [M+H] + (LCMS).

[0146] Example 8. Synthesis of Compound 8: Bis(3-pentyloctyl) 9-(((tetrahydrofuran-2-yl)methyl)amino)heptadecanedioate [ka] Compound 8 was prepared using tetraethyl 8-oxopentadecane-1,7,9,15-tetracarboxylate, 9-oxoheptadecanedioic acid and bis(3-pentyloctyl)9-oxoheptadecanedioate according to the protocol described for compound 6, with the following additional steps: Sodium triacetoxyhydroborate (43.0 mg, 0.20 mmol) was added in one portion to a stirred solution of (tetrahydrofuran-2-yl)methanamine (0.020 mL, 0.20 mmol), acetic acid (0.012 mL, 0.22 mmol) and bis(3-pentyloctyl)9-oxoheptadecanedioate (49.2 mg, 0.07 mmol) in 1,2-DCE (2 mL) and NMP (0.5 mL) at 25° C. under argon. The resulting solution was stirred at 25° C. for 30 h. The reaction mixture was diluted with DCM (15 mL) and water (5 mL) along with Na2CO3 (10 mL). The layers were separated and the aqueous layer was extracted with DCM (3x15 mL). The combined organic layers were dried over MgSO4, filtered and concentrated to dryness under reduced pressure to give the crude product. The resulting residue was purified by flash silica chromatography, elution gradient 0 to 35% 20% MeOH / DCM (with 1% NH4OH) in DCM. The product fractions were concentrated to dryness under reduced pressure to give compound 9 (22.60 mg, 40.8%) as a colorless oil. 1 H NMR(500MHz, methanol-d4)δ ppm 0.9(t,J=6.9Hz,12H)1.2-1.4(m,49H)1.4-1.5(m,6H)1.5-1.7(m,9H)1.9- 2.0(m,2H)2.0-2.1(m,1H)2.3(s,4H)2.5-2.6(m,2H)2.7-2.8(m,1H)3.8(br d,J=7.3Hz,1H)3.8(br d,J=7.6Hz,1H)4.0(br d,J=5.6Hz,1H)4.1-4.1(m,4H);C 48 H 93 NO5 m / z calculated value 763.705 Measured value 764.9 [M+H] + (LCMS).

[0147] Example 9. Synthesis of Compound 9: Bis(3-pentyloctyl) 9-(((1,4-dioxan-2-yl)methyl)amino)heptadecanedioate [ka] Compound 9 was prepared using tetraethyl 8-oxopentadecane-1,7,9,15-tetracarboxylate, 9-oxoheptadecanedioic acid and bis(3-pentyloctyl) 9-oxoheptadecanedioate according to the protocol described for compound 6, with the following additional steps: Sodium triacetoxyhydroborate (36.6 mg, 0.17 mmol) was added in one portion to a stirred solution of (1,4-dioxan-2-yl)methanamine (0.017 mL, 0.16 mmol), acetic acid (9.88 μl, 0.17 mmol) and bis(3-pentyloctyl) 9-oxoheptadecanedioate (39.1 mg, 0.06 mmol) in 1,2-DCE (2 mL) and NMP (0.5 mL) at 25° C. under argon. The resulting solution was stirred at 25° C. for 30 h. The reaction mixture was diluted with DCM (15 mL) and water (5 mL) along with saturated Na2CO3 (10 mL). The layers were separated and the aqueous layer was extracted with DCM (3x15 mL). The combined organic layers were dried over MgSO4, filtered and concentrated to dryness under reduced pressure to give the crude product. The resulting residue was purified by flash silica chromatography, elution gradient 0 to 35% 20% MeOH / DCM (with 1% NH4OH) in DCM. The product fractions were concentrated to dryness under reduced pressure to give compound 9 (23.10 mg, 51.4%) as a colorless oil. 1 H NMR(500MHz, methanol-d4)δ ppm 0.9(t,J=7.1Hz,12H)1.3-1.4(m,48H)1.4-1.5(m,6H)1.6-1.6(m,8H)2.3(t,J=7.3Hz,4H)2.5 -2.6(m,2H)2.6(s,3H)3.3-3.3(m,1H)3.6(s,1H)3.6-3.8(m,4H)3.8-3.8(m,1H)4.1(s,4H);C 48 H 93 NO6 m / z calculated value 779.700 Measured value 780.9 [M+H] + (LCMS).

[0148] Scheme 3 below illustrates the synthetic procedure for preparing Examples 10-12. In step g below, when the respective amine was utilized as the free base, AcOH was used as an additive in the reductive amination.

[0149] Scheme 3: [ka] Reagents:e,f)EDC.HCl, DIPEA, DMAP, DCM;g)NaBH(OAc)3, AcOH, 1,2-DCE:NMP(4:1)

[0150] Example 10. Synthesis of Compound 10: 1-(heptadecan-9-yl) 17-nonyl 9-((2-oxaspiro[3.3]heptan-6-yl)amino)heptadecanedioate Step e): To a stirred solution of 9-oxoheptadecanedioic acid (105.6 mg, 0.34 mmol), N-ethyl-N-isopropylpropan-2-amine (176 μl, 1.01 mmol), heptadecan-9-ol (57.5 mg, 0.22 mmol) and N,N-dimethylpyridin-4-amine (8.62 mg, 0.07 mmol) in DCM (6 mL) at 0° C. under argon was added 3-(((ethylimino)methylene)amino)-N,N-dimethylpropan-1-amine hydrochloride (97 mg, 0.50 mmol) in one portion. The resulting solution was stirred at 25° C. for 18 h. The reaction mixture was concentrated to dryness under reduced pressure and redissolved in EtOAc (15 mL), water (5 mL) and 5% citric acid (10 mL). The layers were separated and the aqueous layer was extracted three times with EtOAc (20 mL). The combined organic layers were dried over MgSO4, filtered and concentrated to dryness under reduced pressure to give the crude product. The resulting residue was purified by flash silica chromatography, elution gradient 10 to 40% EtOAc in hexanes. The product fractions were concentrated to dryness under reduced pressure to give 17-(heptadecan-9-yloxy)-9,17-dioxoheptadecanoic acid (0.083 g, 44.4%) as a colorless oil. 1 H NMR (500 MHz, chloroform-d) δ ppm 0.8-0.9 (m, 6H) 1.2-1.3 (m, 36H) 1.5-1.6 (m, 8H) 1.6-1.6 (m, 4H) 2.2-2.4 (m, 8H) 4.8-4.9 (m, 1H).

[0151] Step f): To a stirred solution of nonan-1-ol (124 μl, 0.71 mmol), N,N-dimethylpyridin-4-amine (6.08 mg, 0.05 mmol), 17-(heptadecan-9-yloxy)-9,17-dioxoheptadecanoic acid (131 mg, 0.24 mmol) and N-ethyl-N-isopropylpropan-2-amine (174 μl, 1.00 mmol) in DCM (4.5 mL) under argon at 0° C. was added 3-(((ethylimino)methylene)amino)-N,N-dimethylpropan-1-amine hydrochloride (145 mg, 0.76 mmol) in one portion. The resulting solution was stirred at 25° C. for 18 h. The reaction mixture was diluted with DCM (10 mL) and saturated NH4Cl (10 mL). The layers were separated and the aqueous layer was extracted three times with DCM (15 mL). The combined organic layers were dried over MgSO4, filtered and concentrated to dryness under reduced pressure to give the crude product. The resulting residue was purified by flash silica chromatography, elution gradient 10 to 30% EtOAc in hexanes. The product fractions were concentrated under reduced pressure to give 1-(heptadecan-9-yl) 17-nonyl 9-oxoheptadecanedioate (0.135 g, 84%) as a colorless oil. 1 H NMR(500MHz,chloroform-d)δ ppm 0.9-0.9(m,9H)1.2-1.3(m,26H)1.3(br s,22H)1.5-1.7(m,14H)2.3(q,J=7.3Hz,4H)2.4(t,J=7.5Hz,4H)4.1(t,J=6.8Hz,2H)4.8-4.9(m,1H).

[0152] Step g): [ka] To a stirred solution of 1-(heptadecan-9-yl)17-nonyl 9-oxoheptadecanedioate (45.3 mg, 0.07 mmol) and 2-oxaspiro[3.3]heptan-6-aminium chloride (23.95 mg, 0.16 mmol) in 1,2-DCE (2 mL) and NMP (0.5 mL) under argon was added sodium triacetoxyhydroborate (35.3 mg, 0.17 mmol) in one portion. The resulting solution was stirred at 25° C. for 50 h. The reaction mixture was diluted with DCM (10 mL), water (5 mL) and saturated Na2CO3 (5 mL). The layers were separated and the aqueous layer was extracted three times with DCM (15 mL). The combined organic layers were dried over MgSO4, filtered and concentrated to dryness under reduced pressure to give the crude product. The resulting residue was purified by flash silica chromatography, elution gradient 0 to 50% 20% MeOH / DCM (with 1% NH4OH) in DCM. The product fractions were concentrated to dryness under reduced pressure to give compound 10 (0.039 g, 75%) as a colorless oil. 1 H NMR(500MHz,methanol-d4)δ ppm 0.9(br t,J=6.5Hz,9H)1.3-1.4(m,56H)1.5-1.6(m,4H)1.6(br d,J=6.1Hz,6H)2.0(br t,J=10.2Hz,2H)2.3(br C 49 H 93 NO5 m / z Calculated value 775.705 Measured value 776.8 [M+H] + (LCMS).

[0153] Example 11. Synthesis of Compound 11: 1-(heptadecan-9-yl) 17-nonyl 9-((7-oxaspiro[3.5]nonan-2-yl)amino)heptadecanedioate [ka] Compound 11 was prepared using 17-(heptadecan-9-yloxy)-9,17-dioxoheptadecanoic acid and 1-(heptadecan-9-yl)17-nonyl 9-oxoheptadecanedioate according to the protocol described for compound 10 with the following additional steps: Sodium triacetoxyhydroborate (34.3 mg, 0.16 mmol) was added in one portion to a stirred solution of 1-(heptadecan-9-yl)17-nonyl 9-oxoheptadecanedioate (43.9 mg, 0.06 mmol) and 7-oxaspiro[3.5]nonane-2-aminium chloride (27.6 mg, 0.16 mmol) in 1,2-DCE (2 mL) and NMP (0.5 mL) under argon. The resulting solution was stirred at 25° C. for 50 h. The reaction mixture was diluted with DCM (10 mL), water (5 mL) and saturated Na2CO3 (5 mL). The layers were separated and the aqueous layer was extracted three times with DCM (15 mL). The combined organic layers were dried over MgSO4, filtered and concentrated to dryness under reduced pressure to give the crude product. The resulting residue was purified by flash silica chromatography, elution gradient 0 to 50% 20% MeOH in DCM MeOH / DCM (with 1% NH4OH). The product fractions were concentrated to dryness under reduced pressure to give compound 11 (0.029 g, 55.4%) as a colorless oil. 1 H NMR(500MHz,methanol-d4)δ ppm 0.9(t,J=6.8Hz,9H)1.3-1.4(m,56H)1.5-1.6(m,8H)1.6(br d,J=5.6Hz,8H)2.2-2.3(m,2H)2.3-2.3(m,4H)2.5(br t,J=5.4Hz,1H)3.3-3.4(m,1H)3.5-3.6(m,2H)3.6-3.7(m,2H)4.1(s,2H)4.9-4.9(m,1H);C 51 H 97 NO5 m / z Calculated value 803.737 Measured value 804.7 [M+H] + (LCMS).

[0154] Example 12. Synthesis of Compound 12: 1-(heptadecan-9-yl) 17-nonyl 9-((tetrahydro-2H-pyran-4-yl)amino)heptadecanedioate [ka] Compound 12 was prepared using 17-(heptadecan-9-yloxy)-9,17-dioxoheptadecanoic acid and 1-(heptadecan-9-yl)17-nonyl 9-oxoheptadecanedioate according to the protocol described for compound 10 with the following additional steps: Sodium triacetoxyhydroborate (30.2 mg, 0.14 mmol) was added in one portion to a stirred solution of tetrahydro-2H-pyran-4-amine (12.65 μl, 0.12 mmol), 1-(heptadecan-9-yl)17-nonyl 9-oxoheptadecanedioate (46.1 mg, 0.07 mmol) and acetic acid (163 μl, 0.16 mmol) in 1,2-DCE (2 mL) and NMP (0.5 mL) under argon. The resulting solution was stirred at 25° C. for 40 h. The reaction mixture was diluted with DCM (10 mL), water (5 mL) and saturated Na2CO3 (5 mL). The layers were separated and the aqueous layer was extracted three times with DCM (15 mL). The combined organic layers were dried over MgSO4, filtered and concentrated to dryness under reduced pressure to give the crude product. The resulting residue was purified by flash silica chromatography, elution gradient 0 to 45% 20% MeOH / DCM (with 1% NH4OH) in DCM. The product fractions were concentrated to dryness under reduced pressure to give compound 12 (0.028 g, 53.8%) as a colorless oil. 1 H NMR(500MHz, methanol-d4)δ ppm 0.9(br t,J=6.3Hz,9H)1.3-1.3(m,27H)1.3-1.5(m,31H)1.5-1.6(m,4H)1.6-1.7(m,6H)1.8(br d,J=12.5Hz,2H)2.3(s,4H)2.7-2.7(m,1H)2.8(br t,J=10.5Hz,1H)3.4(br t,J=11.7Hz,2H)3.9(br d,J=10.8Hz,2H)4.0-4.1(m,2H)4.9-4.9(m,1H);C 48 H 93 NO5 m / z calculated value 763.705 Measured value 764.8 [M+H] + (LCMS).

[0155] Scheme 4 below illustrates the synthetic procedure for preparing Examples 13 and 45. In step k below, when the respective amine was utilized as the free base, AcOH was used as an additive in the reductive amination.

[0156] Scheme 4: [ka] Reagents h)NaOEt, EtOH;i)conc.HCl, AcOH;j)EDC.HCl, DIPEA, DMAP, DCM;k)NaBH(OAc)3, AcOH, 1,2-DCE:NMP(4:1)

[0157] Example 13. Synthesis of Compound 13: Bis(3-pentyloctyl) 7-((2-oxaspiro[3.3]heptan-6-yl)amino)tridecanedioate Step h): To a stirred solution of diethyl 3-oxopentanedioate (1.977 mL, 10.88 mmol) in ethanol (anhydrous, 99.5%) (10 mL) under argon at 81° C., sodium ethanolate (1.111 g, 16.32 mmol) was added in portions. The resulting solution was stirred at 81° C. for 1 h. To it was added ethyl 5-bromopentanoate (3.87 mL, 24.48 mmol) dropwise and stirred at 81° C. for 18 h. The reaction mixture was cooled, the solvent was evaporated, and the reaction mixture was diluted with DCM (25 mL) and washed with saturated aqueous NaCl (25 mL). The organic layer was dried over MgSO4, filtered and concentrated to dryness under reduced pressure to give the crude product. The resulting residue was purified by flash silica chromatography, elution gradient 0 to 60% EtOAc in hexanes. The product fractions were concentrated under reduced pressure to give tetraethyl 6-oxoundecane-1,5,7,11-tetracarboxylate (3.28 g, 65.8%) as a pale yellow liquid. 1 H NMR(500MHz,chloroform-d)δ ppm 1.2-1.4(m,16H)1.6-1.7(m,4H)1.8-2.0(m,4H)2.3-2.4(m,4H)4.1-4.3(m,10H).

[0158] Step i): To a stirred solution of tetraethyl 6-oxoundecane-1,5,7,11-tetracarboxylate (3.821 g, 8.33 mmol) in acetic acid (8.5 mL) at 25° C. was slowly added hydrochloric acid (17.65 mL, 214.99 mmol). The resulting solution was stirred at 102° C. for 18 h with a reflux condenser and a vent to remove excess HCl gas. The reaction was cooled to RT and the reaction mixture was poured onto ice water (50 mL) and allowed to stand for 30 min. The precipitate was collected by filtration, washed with cold water (3×20 mL) and dried under vacuum to give the crude material as a yellow solid. The crude product was purified by crystallization from acetone to give 7-oxotridecanedioic acid (0.283 g, 13.15%) as a white powder. 1 H NMR(500MHz,DMSO-d6)δ ppm 1.2-1.2(m,4H)1.4(m,8H)2.2(t,J=7.3Hz,4H)2.4(t,J=7.3Hz,4H);C 13 H 22 O5 m / z calculated value 258.147 measured value 257.1 [MH] - (LCMS).

[0159] Step j): To a stirred solution of 7-oxotridecanedioic acid (101.8 mg, 0.39 mmol), 3-pentyloctan-1-ol (see WO 2013 / 086354, p191 for procedure to prepare 3-pentyloctan-1-ol) (197 mg, 0.99 mmol), N-ethyl-N-isopropylpropan-2-amine (275 μl, 1.58 mmol) and N,N-dimethylpyridin-4-amine (9.63 mg, 0.08 mmol) in DCM (7 mL) at 0° C. under argon was added 3-(((ethylimino)methylene)amino)-N,N-dimethylpropan-1-amine hydrochloride (204 mg, 1.06 mmol) in one portion. The resulting solution was stirred at 25° C. for 24 h. The reaction mixture was quenched with saturated aqueous NH4Cl (15 mL) and extracted with EtOAc (3x20 mL), and the organic layer was dried over MgSO4, filtered, and concentrated to dryness under reduced pressure to give a colorless oil. The resulting residue was purified by flash silica chromatography, elution gradient 0 to 40% EtOAc in hexanes. The product fractions were concentrated to dryness under reduced pressure to give bis(3-pentyloctyl) 7-oxotridecanedioate (0.227 g, 92%) as a colorless oil. 1 H NMR(500MHz,chloroform-d)δ ppm 0.9(t,J=7.1Hz,12H)1.2-1.4(m,36H)1.4(br d,J=5.0Hz,2H)1.5-1.7(m,12H)2.3(t,J=7.5Hz,4H)2.4(t,J=7.5Hz,4H)4.1(t,J=7.1Hz,4H).

[0160] Step k): [ka] To a stirred solution of bis(3-pentyloctyl) 7-oxotridecanedioate (52 mg, 0.08 mmol) and 2-oxaspiro[3.3]heptan-6-aminium chloride (30.0 mg, 0.20 mmol) in NMP (0.500 mL) and 1,2-DCE (2 mL) at 25 °C under argon was added sodium triacetoxyhydroborate (44.2 mg, 0.21 mmol) in one portion. The resulting suspension was stirred at 25 °C for 40 h. The reaction mixture was diluted with DCM (15 mL), water (5 mL) and saturated Na2CO3 (10 mL). The layers were separated and the aqueous layer was extracted with DCM (3x15 mL). The combined organic layers were dried over MgSO4, filtered and concentrated to dryness under reduced pressure to give the crude product. The resulting residue was purified by flash silica chromatography, elution gradient 0 to 50% 20% MeOH / DCM (with 1% NH4OH) in DCM. The product fractions were concentrated to dryness under reduced pressure to give compound 13 (0.025 g, 41.8%) as a colorless oil. 1 H NMR(500MHz, methanol-d4)δ ppm 0.9-1.0(m,12H)1.3-1.4(m,44H)1.4-1.5(m,2H)1.5-1.7(m,8H)1.9-2.0(m,2H)2.3(t,J= 7.3Hz,4H)2.4-2.6(m,3H)3.2(t,J=7.7Hz,1H)4.1(t,J=6.8Hz,4H)4.6(s,2H)4.7(s,2H);C 48 H 93 NO5 m / z Calculated value 719.643 Measured value 720.7 [M+H] + (LCMS).

[0161] Scheme 5 below illustrates the synthetic procedure for preparing Examples 14-18. In step o below, when the respective amine was utilized as the free base, AcOH was used as an additive in the reductive amination.

[0162] Scheme 5: [ka] Reagents: l) NaOEt, EtOH; m) conc. HCl, AcOH; n) EDC. HCl, DIPEA, DMAP, DCM; o) NaBH(OAc)3, AcOH, 1,2-DCE:NMP (4:1)

[0163] Example 14. Synthesis of Compound 14: Bis(3-pentyloctyl) 10-((2-oxaspiro[3.3]heptan-6-yl)amino)nonadecanedioate Step l): Sodium ethanolate (2.52 g, 37.09 mmol) was added portionwise to a stirred solution of diethyl 3-oxopentanedioate (4.49 mL, 24.73 mmol) in ethanol (anhydrous, 99.5%) (15 mL) at 25° C. under argon over 10 min. The resulting suspension was stirred at 81° C. for 1 h. Ethyl 8-bromooctanoate (12.53 mL, 59.35 mmol) was added dropwise to the reaction mixture and the suspension was stirred at 81° C. for an additional 18 h. The reaction mixture was cooled to RT, concentrated to dryness under reduced pressure, redissolved in DCM (50 mL) and washed successively with water (50 mL), saturated aqueous sodium chloride solution (50 mL). The organic layer was dried over MgSO4, filtered and concentrated under reduced pressure to give the crude product. The resulting residue was purified by flash silica chromatography, elution gradient 0 to 50% EtOAc in hexanes. The product fractions were concentrated to dryness under reduced pressure to give tetraethyl 9-oxoheptadecane-1,8,10,17-tetracarboxylate (8.80 g, 65.6%) as a pale yellow oil. 1 H NMR(500MHz,chloroform-d)δ ppm 1.2-1.4(m,28H)1.6-1.7(m,4H)1.7-1.9(m,4H)2.2-2.3(m,4H)4.1-4.2(m,10H).

[0164] Step m): To a stirred solution of tetraethyl 9-oxoheptadecane-1,8,10,17-tetracarboxylate (8.8 g, 16.21 mmol) in acetic acid (21 mL) at 25° C., hydrochloric acid (34.4 mL, 418.34 mmol) was added slowly. The resulting solution was stirred at 102° C. for 18 h with a reflux condenser and a vent to remove excess HCl gas. The reaction was cooled to RT and the reaction mixture was poured onto ice water (40 mL) and allowed to stand for 30 min. The precipitate was collected by filtration, washed with cold water (3×20 mL), and dried under vacuum to give the crude product as a light yellow solid. The crude product was purified by crystallization from acetone to give 10-oxononadecanedioic acid (1.963 g, 35.3%). 1 H NMR(500MHz,DMSO-d6)δ ppm 1.2(br s,16H)1.4-1.5(m,8H)2.2(t,J=7.3Hz,4H)2.3-2.4(m,4H)12.0(br s,2H);C 19 H 34 O5 m / z calculated value 342.241 measured value 341.2 [MH] - (LCMS).

[0165] Step n): To a stirred solution of 10-oxononadecanedioic acid (180 mg, 0.53 mmol), N,N-dimethylpyridin-4-amine (9.63 mg, 0.08 mmol), 3-pentyloctan-1-ol (see WO 2013 / 086354, p191 for procedure to prepare 3-pentyloctan-1-ol) (253 mg, 1.26 mmol) and N-ethyl-N-isopropylpropan-2-amine (330 μl, 1.89 mmol) in DCM (8 mL) at 0° C. under argon was added 3-(((ethylimino)methylene)amino)-N,N-dimethylpropan-1-amine hydrochloride (272 mg, 1.42 mmol) in one portion. The resulting solution was stirred at 25° C. for 18 h. The reaction mixture was diluted with DCM (15 mL) and water (15 mL). The layers were separated and the aqueous layer was extracted with DCM (3x20 mL). The combined organic layers were washed with 0.5 M citric acid (15 mL) and saturated aqueous NaCl (15 mL). The organic layers were dried over MgSO4, filtered and concentrated to dryness under reduced pressure to give the crude product. The resulting residue was purified by flash silica chromatography, elution gradient 0 to 30% hexanes in EtOAc. The product fractions were concentrated to dryness under reduced pressure to give bis(3-pentyloctyl) 10-oxononadecandioate (0.343 g, 92%) as a colorless oil. 1 H NMR(500MHz,chloroform-d)δ ppm 0.9(t,J=7.1Hz,12H)1.2-1.4(m,48H)1.4(br s,2H)1.5-1.6(m,12H)2.3(t,J=7.5Hz,4H)2.3-2.4(m,4H)4.0-4.1(m,4H).

[0166] Step o): [ka] To a stirred solution of bis(3-pentyloctyl) 10-oxononadecanedioate (75.7 mg, 0.11 mmol) and 2-oxaspiro[3.3]heptan-6-aminium chloride (33.6 mg, 0.22 mmol) in 1,2-DCE (2.4 mL) and NMP (0.6 mL) at 25 °C under argon was added sodium triacetoxyhydroborate (54.4 mg, 0.26 mmol) in one portion. The resulting suspension was stirred at 25 °C for 40 h. The reaction mixture was diluted with DCM (10 mL), water (5 mL) and saturated Na2CO3 (5 mL). The layers were separated and the aqueous layer was extracted with DCM (3x10 mL). The combined organic layers were dried over MgSO4, filtered and concentrated to dryness under reduced pressure to give the crude product. The resulting residue was purified by flash silica chromatography, elution gradient 0 to 35% 20% MeOH / DCM (with 1% NH4OH) in DCM. The product fractions were concentrated to dryness under reduced pressure to give compound 14 (0.033 g, 38.0%) as a colorless oil. 1 H NMR(500MHz, methanol-d4)δ ppm 0.9(t,J=7.0Hz,12H)1.2-1.4(m,56H)1.4-1.5(m,2H)1.5-1.6(m,8H)2.0(td,J=9.1,2.8Hz,2H)2.3(t,J=7.3Hz,4H)2.4-2.6(m,3H)3.2(br t,J=7.8Hz,1H)4.1(t,J=6.7Hz,4H)4.6(s,2H)4.7(s,1H);C 51 H 97 NO5 m / z Calculated value 803.737 Measured value 804.8 [M+H] + (LCMS).

[0167] Example 15. Synthesis of Compound 15: Bis(3-pentyloctyl) 10-((tetrahydro-2H-pyran-4-yl)amino)nonadecanedioate [ka] Compound 15 was prepared using tetraethyl 9-oxoheptadecane-1,8,10,17-tetracarboxylate, 10-oxononadecanedioic acid and bis(3-pentyloctyl)10-oxononadecanedioate according to the protocol described for compound 14, with the following additional steps: Sodium triacetoxyhydroborate (75 mg, 0.36 mmol) was added in one portion to a stirred solution of bis(3-pentyloctyl)10-oxononadecanedioate (93 mg, 0.13 mmol), acetic acid (22.56 μl, 0.39 mmol) and tetrahydro-2H-pyran-4-amine (32.7 μl, 0.32 mmol) in 1,2-DCE (2.4 mL) and NMP (0.6 mL) at 25° C. under argon. The resulting suspension was stirred at 25° C. for 50 h. The reaction mixture was diluted with DCM (10 mL), water (5 mL) and saturated Na2CO3 (5 mL). The layers were separated and the aqueous layer was extracted with DCM (3x10 mL). The combined organic layers were dried over MgSO4, filtered and concentrated to dryness under reduced pressure to give the crude product. The resulting residue was purified by flash silica chromatography, elution gradient 0 to 35% 20% MeOH / DCM (with 1% NH4OH) in DCM. The product fractions were concentrated to dryness under reduced pressure to give compound 15 (0.036 g, 34.4%) as a colorless oil. 1 H NMR(500MHz, methanol-d4)δ ppm 0.9(t,J=7.0Hz,12H)1.2-1.3(m,32H)1.3-1.4(m,20H)1.4-1.5(m,8H)1.5-1.7(m,8H)1.9(br C 50 H 97 NO5 m / z Calculated value 791.737 Measured value 792.8 [M+H] + (LCMS).

[0168] Example 16. Synthesis of Compound 16: Bis(3-pentyloctyl) 10-(((tetrahydro-2H-pyran-2-yl)methyl)amino)nonadecanedioate [ka] Compound 16 was prepared using tetraethyl 9-oxoheptadecane-1,8,10,17-tetracarboxylate, 10-oxononadecanedioic acid and bis(3-pentyloctyl)10-oxononadecanedioate according to the protocol described for compound 14, with the following additional steps: Sodium triacetoxyhydroborate (41.0 mg, 0.19 mmol) was added in one portion to a stirred solution of bis(3-pentyloctyl)10-oxononadecanedioate (50.7 mg, 0.07 mmol) and (tetrahydro-2H-pyran-2-yl)methanaminium chloride (27.2 mg, 0.18 mmol) in 1,2-DCE (2.0 mL) and NMP (0.6 mL) at 25° C. under argon. The resulting suspension was stirred at 25° C. for 50 h. The reaction mixture was diluted with DCM (10 mL), water (5 mL) and saturated Na2CO3 (5 mL). The layers were separated and the aqueous layer was extracted with DCM (5x10 mL). The combined organic layers were dried over MgSO4, filtered and concentrated to dryness under reduced pressure to give the crude product. The resulting residue was purified by flash silica chromatography, elution gradient 0 to 35% 20% MeOH / DCM (with 1% NH4OH) in DCM. The product fractions were concentrated to dryness under reduced pressure to give compound 16 (26.9 mg, 46.5%) as a colorless oil. 1 H NMR(400MHz, methanol-d4)δ ppm 0.9(t,J=7.0Hz,12H)1.3(br d,J=13.8Hz,54H)1.4-1.5(m,6H)1.5-1.7(m,12H)1.8-1.9(m,1H)2.3(s,4H)2 .5-2.6(m,2H)2.6-2.7(m,1H)3.4-3.5(m,2H)3.9-4.0(m,1H)4.1-4.1(m,4H);C 51 H 99 NO5 m / z Calculated value 805.752 Measured value 807.0 [M+H] + (LCMS).

[0169] Example 17. Synthesis of Compound 17: Bis(3-pentyloctyl) 10-(((tetrahydro-2H-pyran-4-yl)methyl)amino)nonadecanedioate [ka] Compound 17 was prepared using tetraethyl 9-oxoheptadecane-1,8,10,17-tetracarboxylate, 10-oxononadecanedioic acid and bis(3-pentyloctyl)10-oxononadecanedioate according to the protocol described for compound 14, with the following additional steps: Sodium triacetoxyhydroborate (43.0 mg, 0.20 mmol) was added in one portion to a stirred solution of bis(3-pentyloctyl)10-oxononadecanedioate (53.2 mg, 0.08 mmol), acetic acid (12.91 μl, 0.23 mmol) and (tetrahydro-2H-pyran-4-yl)methanamine (17.84 μl, 0.16 mmol) in 1,2-DCE (2 mL) and NMP (0.5 mL) under nitrogen. The resulting solution was stirred at 25° C. for 45 h. The reaction mixture was diluted with DCM (10 mL), water (5 mL) and saturated Na2CO3 (5 mL). The layers were separated and the aqueous layer was extracted with DCM (3x10 mL). The combined organic layers were dried over MgSO4, filtered and concentrated to dryness under reduced pressure to give the crude product. The resulting residue was purified by flash silica chromatography, elution gradient 0 to 35% 20% MeOH / DCM (with 1% NH4OH) in DCM. The product fractions were concentrated to dryness under reduced pressure to give compound 17 (28 mg, 45.7%) as a colorless oil. 1 H NMR(500MHz, methanol-d4)δ ppm 0.9-0.9(m,12H)1.2-1.4(m,54H)1.4-1.5(m,6H)1.6-1.6(m,8H)1.7-1.8(m,3H)2.3(t,J=7.3Hz,4H)2.5-2.6(m,3H)3.4-3.5(m,2H)3.9(br dd,J=11.2,4.0Hz,2H)4.1(t,J=6.7Hz,4H);C 51 H 99 NO5 m / z Calculated value 805.752 Measured value 806.7 [M+H] + (LCMS).

[0170] Example 18. Synthesis of Compound 18: Bis(3-pentyloctyl) 10-((oxetan-3-ylmethyl)amino)nonadecanedioate [ka] Compound 18 was prepared using tetraethyl 9-oxoheptadecane-1,8,10,17-tetracarboxylate, 10-oxononadecanedioic acid, and bis(3-pentyloctyl)10-oxononadecanedioate according to the protocol described for compound 14, with the following additional steps: Sodium triacetoxyhydroborate (31.6 mg, 0.15 mmol) was added in one portion to a stirred solution of oxetan-3-ylmethanaminium chloride (16.40 mg, 0.13 mmol) and bis(3-pentyloctyl)10-oxononadecanedioate (39.1 mg, 0.06 mmol) in 1,2-DCE (2 mL) and NMP (0.5 mL) under nitrogen at 25° C. The resulting solution was stirred at 25° C. for 40 h. The reaction mixture was diluted with DCM (10 mL), water (5 mL) and saturated Na2CO3 (5 mL). The layers were separated and the aqueous layer was extracted with DCM (3x10 mL). The combined organic layers were dried over MgSO4, filtered and concentrated to dryness under reduced pressure to give the crude product. The resulting residue was purified by flash silica chromatography, elution gradient 0 to 50% 20% MeOH / DCM (with 1% NH4OH) in DCM. The product fractions were concentrated to dryness under reduced pressure to give compound 18 (0.017 g, 40.4%) as a colorless oil. 1 H NMR(500MHz, methanol-d4)δ ppm 0.9(t,J=6.9Hz,12H)1.3(br d,J=18.2Hz,52H)1.4-1.5(m,6H)1.5-1.7(m,8H)2.3(t,J=7.2Hz,4H)2.5(br C 49 H 95 NO5 m / z calculated value 777.721 measured value 778.8 [M+H] + (LCMS).

[0171] Scheme 6 below illustrates the synthetic procedure for preparing Examples 19 and 20. In step r below, when the respective amine was utilized as the free base, AcOH was used as an additive in the reductive amination.

[0172] Scheme 6: [ka] Reagents: p, q) EDC.HCl, DIPEA, DMAP, DCM; r) NaBH(OAc)3, AcOH, 1,2-DCE:NMP (4:1)

[0173] Example 19. Synthesis of Compound 19: 1-(heptadecan-9-yl) 13-nonyl 7-((2-oxaspiro[3.3]heptan-6-yl)amino) tridecane dioate Step p): To a stirred solution of 7-oxotridecanedioic acid (181.4 mg, 0.70 mmol), N-ethyl-N-isopropylpropan-2-amine (367 μl, 2.11 mmol), heptadecan-9-ol (120 mg, 0.47 mmol) and N,N-dimethylpyridin-4-amine (18.02 mg, 0.15 mmol) in DCM (9 mL) at 0° C. under argon was added 3-(((ethylimino)methylene)amino)-N,N-dimethylpropan-1-amine hydrochloride (202 mg, 1.05 mmol) in one portion. The resulting solution was stirred at 25° C. for 18 h. The reaction mixture was concentrated to dryness under reduced pressure and redissolved in EtOAc (15 mL), water (10 mL) and 5% citric acid solution (5 mL). The layers were separated and the aqueous layer was extracted with EtOAc (3×20 mL). The combined organic layers were dried over MgSO4, filtered and concentrated to dryness under reduced pressure to give the crude product. The resulting residue was purified by flash silica chromatography, elution gradient 0 to 40% EtOAc in hexanes. The product fractions were concentrated to dryness under reduced pressure to give 13-(heptadecan-9-yloxy)-7,13-dioxotridecanoic acid (0.102 g, 29.2%) as a colorless oil. 1 H NMR(500MHz,chloroform-d)δ ppm C 30 H 56 O5 m / z calculated 496.413, observed 495.5[MH]-+(LCMS).

[0174] Step q): To a stirred solution of 13-(heptadecan-9-yloxy)-7,13-dioxotridecanoic acid (102 mg, 0.21 mmol), nonan-1-ol (64.2 μl, 0.37 mmol), N,N-dimethylpyridin-4-amine (5.02 mg, 0.04 mmol) and N-ethyl-N-isopropylpropan-2-amine (118 μl, 0.68 mmol) in DCM (6 mL) at 0° C. under nitrogen was added 3-(((ethylimino)methylene)amino)-N,N-dimethylpropan-1-amine hydrochloride (83 mg, 0.43 mmol) in one portion. The resulting solution was stirred at 25° C. for 30 h and allowed to come to RT. The reaction mixture was diluted with DCM (15 mL) and water (15 mL). The layers were separated and the aqueous layer was extracted with DCM (3×15 mL). The combined organic layers were washed with 5% citric acid solution (10 mL). The organic layer was dried over MgSO4, filtered and concentrated to dryness under reduced pressure to give the crude product. The resulting residue was purified by flash silica chromatography, elution gradient 0 to 40% EtOAc in hexanes. The product fractions were concentrated to dryness under reduced pressure to give 1-(heptadecan-9-yl) 13-nonyl 7-oxotridecanedioate (0.085 g, 66.5%) as a colorless oil. 1 H NMR(500MHz,chloroform-d)δ ppm 0.9(t,J=6.9Hz,9H)1.2-1.4(m,40H)1.5-1.6(m,4H)1.6-1.7(m,10H)2.3 (q,J=7.1Hz,4H)2.4(t,J=7.4Hz,4H)4.1(t,J=6.7Hz,2H)4.8-4.9(m,1H).

[0175] Step r): [ka] To a stirred suspension of 1-(heptadecan-9-yl)13-nonyl 7-oxotridecanedioate (40.9 mg, 0.07 mmol) and 2-oxaspiro[3.3]heptan-6-aminium chloride (23.57 mg, 0.16 mmol) in 1,2-DCE (2 mL) and NMP (0.5 mL) under nitrogen was added sodium triacetoxyhydroborate (37.6 mg, 0.18 mmol) in one portion. The resulting suspension was stirred at 25° C. for 40 h. The reaction mixture was diluted with DCM (15 mL), water (5 mL) and saturated Na2CO3 (10 mL). The layers were separated and the aqueous layer was extracted with DCM (3×15 mL). The combined organic layers were dried over MgSO4, filtered and concentrated to dryness under reduced pressure to give the crude product. The resulting residue was purified by flash silica chromatography, elution gradient 0 to 50% 20% MeOH / DCM (with 1% NH4OH) in DCM. The product fractions were concentrated to dryness under reduced pressure to give compound 19 (0.023 g, 49.3%) as a colorless oil. 1 H NMR(500MHz, methanol-d4)δ ppm 0.9(t,J=6.6Hz,9H)1.3-1.4(m,48H)1.5-1.6(m,4H)1.6-1.7(m,6H)1.9-2.0(m,2H)2.3(br C 45 H85NO5 m / z Calculated value 719.643 Measured value 720.8 [M+H] + (LCMS).

[0176] Example 20. Synthesis of Compound 20: 1-(heptadecan-9-yl) 13-nonyl 7-((oxetan-3-ylmethyl)amino) tridecane dioate [ka] Compound 20 was prepared using 13-(heptadecan-9-yloxy)-7,13-dioxotridecanoic acid and 1-(heptadecan-9-yl)13-nonyl 7-oxotridecanedioate according to the protocol described for compound 19 with the following additional steps: Sodium triacetoxyhydroborate (40.6 mg, 0.19 mmol) was added in one portion to a stirred suspension of 1-(heptadecan-9-yl)13-nonyl 7-oxotridecanedioate (44.2 mg, 0.07 mmol) and oxetan-3-ylmethanaminium chloride (21.04 mg, 0.17 mmol) in 1,2-DCE (2 mL) and NMP (0.5 mL) under nitrogen. The resulting suspension was stirred at 25° C. for 40 h. The reaction mixture was diluted with DCM (15 mL), water (5 mL) and saturated Na2CO3 (10 mL). The layers were separated and the aqueous layer was extracted with DCM (3x15 mL). The combined organic layers were dried over MgSO4, filtered and concentrated to dryness under reduced pressure to give the crude product. The resulting residue was purified by flash silica chromatography, elution gradient 0 to 35% 20% MeOH / DCM (with 1% NH4OH) in DCM. The product fractions were concentrated to dryness under reduced pressure to give compound 20 (0.023 g, 46.3%) as a colorless oil. 1 H NMR(500MHz, methanol-d4)δ ppm 0.9(s,9H)1.3-1.4(m,44H)1.4-1.5(m,4H)1.5-1.6(m,4H)1.6-1.7(m,6H)2.3(br t,J=7.2Hz,4H)2.5(br C 43 H 83 NO5 m / z Calculated value 693.627 Measured value 694.8 [M+H] + (LCMS).

[0177] Scheme 7 below illustrates the synthetic procedure for preparing Examples 21 and 22. In step t below, when the respective amine was utilized as the free base, AcOH was used as an additive in the reductive amination.

[0178] Scheme 7: [ka] Reagents s)EDC.HCl, DIPEA, DMAP, DCM; t)NaBH(OAc)3, AcOH, 1,2-DCE:NMP(4:1)

[0179] Example 21. Synthesis of Compound 21: 1-(heptadecan-9-yl) 17-(2-methylnonyl) 9-(((tetrahydrofuran-3-yl)methyl)amino)heptadecanedioate Step s): To a stirred solution of 2-methylnonan-1-ol (52.5 mg, 0.33 mmol), N-ethyl-N-isopropylpropan-2-amine (0.104 mL, 0.60 mmol), N,N-dimethylpyridin-4-amine (3.04 mg, 0.02 mmol) and 17-(heptadecan-9-yloxy)-9,17-dioxoheptadecanoic acid (91.7 mg, 0.17 mmol) in DCM (4 mL) at 0° C. under argon was added 3-(((ethylimino)methylene)amino)-N,N-dimethylpropan-1-amine hydrochloride (66.8 mg, 0.35 mmol) in one portion. The resulting solution was stirred at 25° C. for 16 h. The reaction mixture was diluted with DCM (20 mL), 5% citric acid solution (20 mL). The layers were separated and the aqueous layer was extracted with DCM (3x20 mL). The combined organic layers were washed with saturated aqueous NaCl (20 mL). The organic layers were dried over MgSO4, filtered and concentrated to dryness under reduced pressure to give the crude product. The resulting residue was purified by flash silica chromatography, elution gradient 0 to 35% EtOAc in hexanes. The product fractions were concentrated to dryness under reduced pressure to give 1-(heptadecan-9-yl) 17-(2-methylnonyl) 9-oxoheptadecanedioate (87 mg, 76%) as a colorless oil. 1 H NMR(500MHz,chloroform-d)δ ppm 0.8-0.9(m,12H)1.2-1.4(m,48H)1.4-1.7(m,13H)2.3(dt,J=12.4,7.5Hz,4H)2.4(t,J=7.5Hz,4H)3.8-4.0(m,2H)4.8-4.9(m,1H).

[0180] Step t): [ka] (After 10 min) sodium triacetoxyhydroborate (29.8 mg, 0.14 mmol) was added in one portion to a stirred solution of 1-(heptadecan-9-yl)17-(2-methylnonyl)9-oxoheptadecanedioate (39 mg, 0.06 mmol) and (tetrahydrofuran-3-yl)methanamine (0.014 mL, 0.14 mmol) in 1,2-DCE (2 mL) and NMP (0.5 mL) under argon. The resulting solution was stirred at 25° C. for 18 h. The reaction mixture was diluted with DCM (15 mL), water (5 mL) and saturated Na2CO3 (10 mL). The layers were separated and the aqueous layer was extracted with DCM (3×15 mL). The combined organic phase was dried over MgSO4, filtered and evaporated to dryness to give the crude product. The resulting residue was purified by flash silica chromatography, elution gradient 0 to 40% 20% MeOH in DCM (with 1% NH4OH) in DCM. The product fractions were concentrated to dryness under reduced pressure to give compound 21 (30.3 mg, 69.2%) as a colorless oil. 1 H NMR(500MHz, methanol-d4)δ ppm 0.9-1.0(m,12H)1.1-1.2(m,1H)1.2-1.4(m,52H)1.4-1.5(m,4H)1.5-1.6(m,4H)1.6-1.7(m,5H)1.7-1.8(m,1H)2.1-2.2(m,1 H)2.3-2.4(m,4H)2.4-2.5(m,1H)2.6-2.7(m,3H)3.5(dd,J=8.5,6.2Hz,1H)3.7-3.8(m,1H)3.8-4.0(m,4H)4.9-4.9(m,1H);C 49 H 95 NO5 m / z calculated value 777.721 measured value 778.8 [M+H] + (LCMS).

[0181] Example 22. Synthesis of Compound 22: 1-(heptadecan-9-yl) 17-(2-methylnonyl) 9-((2-oxaspiro[3.3]heptan-6-yl)amino)heptadecanedioate [ka] Compound 22 was prepared using 1-(heptadecan-9-yl) 17-(2-methylnonyl) 9-oxoheptadecanedioate according to the protocol described for compound 21 with the following additional steps: (After 10 min) sodium triacetoxyhydroborate (30.7 mg, 0.14 mmol) was added in one portion to a stirred solution of 1-(heptadecan-9-yl) 17-(2-methylnonyl) 9-oxoheptadecanedioate (40.2 mg, 0.06 mmol) and 2-oxaspiro[3.3]heptan-6-aminium chloride (20.83 mg, 0.14 mmol) in 1,2-DCE (2 mL) and NMP (0.5 mL) under argon. The resulting solution was stirred at 25° C. for 18 h. The reaction mixture was diluted with DCM (15 mL), water (5 mL) and saturated Na2CO3 (10 mL). The layers were separated and the aqueous layer was extracted with DCM (3x15 mL). The combined organic layers were dried over MgSO4, filtered and evaporated to dryness to give the crude product. The resulting residue was purified by flash silica chromatography, elution gradient 0 to 40% 20% MeOH in DCM (with 1% NH4OH) in DCM. The product fractions were concentrated to dryness under reduced pressure to give compound 22 (12.10 mg, 26.4%) as a colorless oil. 1 H NMR(500MHz, methanol-d4)δ ppm 0.9-1.0(m,12H)1.3-1.4(m,56H)1.5-1.6(m,4H)1.6-1.7(m,4H)1.7-1.8(m,1H)2.0-2.0(m,2H)2.3-2. 3(m,4H)2.5-2.6(m,3H)3.2-3.2(m,1H)3.9-4.0(m,2H)4.6-4.6(m,2H)4.7-4.7(m,2H)4.9-4.9(m,1H);C 50 H 95 NO5 m / z calculated 789.721, found 790.8 [M+H]+ (LCMS).

[0182] Scheme 8 below illustrates the synthetic procedure for preparing Examples 23 and 24.

[0183] Scheme 8: [ka] Reagents: a, b) EDC.HCl, DIPEA, DMAP, DCM; c, d) NaBH(OAc)3,1,2-DCE:NMP(4:1)

[0184] Example 23. Synthesis of Compound 23: Di(heptadecan-9-yl) 8-((2-oxaspiro[3.3]heptan-6-yl)amino)pentadecanedioate Step 1: Di(heptadecan-9-yl) 8-oxopentadecane dioate and 15-(heptadecan-9-yloxy)-8,15-dioxopentadecanoic acid To a stirred solution of 9-heptadecanol (0.143 g, 0.56 mmol), 8-oxopentadecanedioic acid (0.200 g, 0.70 mmol), DIPEA (0.378 mL, 2.17 mmol) and DMAP (0.017 g, 0.14 mmol) in DCM (5 mL) at 0° C. under argon was added EDC (0.201 g, 1.05 mmol) in one portion. The resulting solution was stirred at room temperature for 16 h. The reaction mixture was concentrated to dryness under reduced pressure and redissolved in EtOAc (20 mL) and sodium biocarbonate (20 mL). The layers were separated and the aqueous layer was extracted with EtOAc (3×30 mL). The combined organic layers were washed successively with 5% citric acid (25 mL) and saturated aqueous NaCl (25 mL). The organic layer was dried over MgSO4, filtered and concentrated to dryness under reduced pressure to give the crude product. The resulting residue was purified by flash silica chromatography, elution gradient 0 to 60% EtOAc in hexanes. The product fractions were concentrated to dryness under reduced pressure to give di(heptadecan-9-yl) 8-oxopentadecane dioate (0.064 g, 11.99%) and 15-(heptadecan-9-yloxy)-8,15-dioxopentadecane acid (0.102 g, 27.8%) as colorless oils. Di(heptadecan-9-yl) 8-oxopentadecane dioate: 1 H NMR (500 MHz, chloroform-d, 22° C.) δ ppm 0.88 (12H,t), 1.26 (54H,m), 1.57 (18H,m), 2.24-2.31 (4H,t), 2.34-2.44 (4H,t), 4.87 (2H,m). 15-(heptadecan-9-yloxy)-8,15-dioxopentadecanoic acid: 1 H NMR (500 MHz, chloroform-d, 22° C.) δ ppm 0.84-0.94 (6H, t), 1.20-1.70 (44H, m), 2.24-2.45 (8H, m), 4.87 (1H, m).

[0185] Step 2: 1-(heptadecan-9-yl) 15-nonyl 8-oxopentadecane dioate To a stirred solution of 15-(heptadecan-9-yloxy)-8,15-dioxopentadecanoic acid (0.071 g, 0.14 mmol)), nonan-1-ol (0.035 mL, 0.20 mmol), DIPEA (0.073 mL, 0.42 mmol) and DMAP (3.31 mg, 0.03 mmol) in DCM (5 mL) at 0° C. under argon was added EDC (0.054 g, 0.28 mmol) in one portion. The resulting solution was stirred at room temperature for 16 h. The reaction mixture was concentrated to dryness under reduced pressure and redissolved in EtOAc (20 mL) and sodium biocarbonate (20 mL). The layers were separated and the aqueous layer was extracted with (EtOAc) (3×30 mL). The organic layer was dried over MgSO4, filtered and concentrated to dryness under reduced pressure to give the crude product. The resulting residue was purified by flash silica chromatography, elution gradient 0 to 60% EtOAc in hexanes. The product fractions were concentrated to dryness under reduced pressure to give 1-(heptadecan-9-yl)15-nonyl 8-oxopentadecanedioate (0.063 g, 71.5%) as a colorless oil. 1 H NMR (500 MHz, chloroform-d, 27° C.) δ ppm 0.83-0.94 (9H, m), 1.21-1.68 (58H, m), 2.22-2.33 (4H, m), 2.38 (4H, t), 4.06 (2H, t), 4.87 (1H, m).

[0186] Compound 23: [ka] To a stirred solution of 2-oxaspiro[3.3]heptan-6-amine hydrochloride (0.038 g, 0.25 mmol) and di(heptadecan-9-yl)8-oxopentadecandioate (0.0639 g, 0.08 mmol) in DCE (4 mL) and NMP (1 mL) at 0° C. under argon was added sodium triacetoxyborohydride (0.055 g, 0.26 mmol) in one portion. The resulting solution was stirred at room temperature for 16 h. The reaction mixture was diluted with DCM (50 mL) and saturated sodium carbonate (50 mL). The layers were separated and the aqueous layer was extracted with (DCM) (3×25 mL). The organic layer was dried over MgSO4, filtered and concentrated to dryness under reduced pressure to give the crude product. The resulting residue was purified by flash silica chromatography, elution gradient 0 to 100% in DCM (20% MeOH and 1% NH4OH in DCM). The product fractions were concentrated to dryness under reduced pressure to give di(heptadecan-9-yl) 8-((2-oxaspiro[3.3]heptan-6-yl)amino)pentadecanedioate (0.046 g, 64.3%) as a colorless oil. 1 H NMR (500MHz, chloroform-d, 22℃) δ ppm 0.83-0.94(12H,m),1.19-1.39(62H,m),1.51(10H,m),1.58-1.66(4H,m),1.83(2H,m),2.28(4H,t),2.37- 2.44(1H,m),2.51-2.58(2H,m),3.03-3.20(1H,m),4.57-4.62(2H,s),4.71(2H,s),4.87(2H,m);C44H87NO5 m / z Calculated value 859.799 Actual value 861.0[M+H]+(LCMS).

[0187] Example 24. Synthesis of Compound 24: 1-(heptadecan-9-yl) 15-nonyl 8-((2-oxaspiro[3.3]heptan-6-yl)amino)pentadecanedioate [ka] Compound 24 was prepared using 15-(heptadecan-9-yloxy)-8,15-dioxopentadecanoic acid and 1-(heptadecan-9-yl)15-nonyl 8-oxopentadecandioate according to the protocol described for compound 23 with the following additional steps: Sodium triacetoxyborohydride (0.062 g, 0.29 mmol) was added in one portion to a stirred solution of 2-oxaspiro[3.3]heptan-6-amine hydrochloride (0.043 g, 0.29 mmol) and 1-(heptadecan-9-yl)15-nonyl 8-oxopentadecandioate (0.063 g, 0.10 mmol) in DCE (4 mL) and NMP (1 mL) at 0° C. under argon. The resulting solution was stirred at room temperature for 16 h. The reaction mixture was diluted with DCM (50 mL) and saturated sodium carbonate (50 mL). The layers were separated and the aqueous layer was extracted with (DCM) (3x25mL). The organic layer was dried over MgSO4, filtered and concentrated to dryness under reduced pressure to give the crude product. The resulting residue was purified by flash silica chromatography, elution gradient 0 to 100% in DCM (20% MeOH and 1% NH4OH in DCM). The product fractions were concentrated to dryness under reduced pressure to give compound 24 (0.036g, 49.7%) as a colorless oil. 1 H NMR (500MHz, chloroform-d, 22℃) δ ppm 0.82-0.96(9H,m),1.28(62H,m),1.81-1.91(2H,m),2.25-2.35(4H,m),2.39-2.48(1H,m),2.52 -2.60(2H,m),3.11-3.20(1H,m),4.08(2H,t),4.62(2H,s),4.73(2H,s),4.89(1H,m);C47H89NO5 m / z Calculated value 747.674 Actual value 748.8[M+H]+(LCMS).

[0188] Scheme 9 below illustrates the synthetic procedure for preparing Example 25.

[0189] Scheme 9: [ka] Reagents: a) EDC.HCl, DIPEA, DMAP, DCM; b) NaBH(OAc)3, 1,2-DCE:NMP(4:1)

[0190] Example 25. Synthesis of Compound 25: 1-(decane-2-yl) 15-(heptadecan-9-yl) 8-((2-oxaspiro[3.3]heptan-6-yl)amino)pentadecanedioate Step 1: 1-(decane-2-yl) 15-(heptadecan-9-yl) 8-oxopentadecane diacetate To a stirred mixture of 15-(heptadecan-9-yloxy)-8,15-dioxopentadecanoic acid (124.1 mg, 0.24 mmol), decan-2-ol (0.055 mL, 0.50 mmol), DIPEA (0.169 mL, 0.97 mmol) and DMAP (5.78 mg, 0.05 mmol) in DCM (5 mL) at 0° C. under argon was added EDC (141 mg, 0.73 mmol) in one portion. The resulting mixture was stirred at room temperature for 16 h. The reaction mixture was diluted with sodium bicarbonate (25 mL) and DCM (25 mL). The layers were separated and the aqueous layer was extracted with (DCM) (4×25 mL). The combined organic layers were dried over MgSO4, filtered and concentrated to dryness under reduced pressure to give the crude product. The resulting residue was purified by flash silica chromatography, elution gradient 0 to 40% EtOAc in hexanes. The product fractions were concentrated to dryness under reduced pressure to give 1-(decan-2-yl) 15-(heptadecan-9-yl) 8-oxopentadecanedioate (87 mg, 55.3%) as a colorless oil. 1 H NMR (500 MHz, chloroform-d, 22° C.) δ ppm 0.89 (9H, t), 1.16-1.70 (61H, m), 2.27 (4H, m), 2.33-2.45 (4H, m), 4.80-4.97 (2H, m).

[0191] Compound 25: [ka] To a stirred solution of 2-oxaspiro[3.3]heptan-6-amine hydrochloride (16.20 mg, 0.11 mmol) and 1-(decane-2-yl)15-(heptadecan-9-yl)8-oxopentadecane-dioate (30 mg, 0.05 mmol) in DCE (2 mL) and NMP (0.5 mL) at 0° C. under argon was added sodium triacetoxyborohydride (25.8 mg, 0.12 mmol) in one portion. The resulting solution was stirred at room temperature for 16 h. The reaction mixture was diluted with DCM (50 mL) and saturated sodium carbonate (50 mL). The layers were separated and the aqueous layer was extracted with (DCM) (3×25 mL). The organic layer was dried over MgSO4, filtered and concentrated to dryness under reduced pressure to give the crude product. The resulting residue was purified by flash silica chromatography, elution gradient 0 to 100% in DCM (20% MeOH and 1% NH4OH in DCM). The product fractions were concentrated to dryness under reduced pressure to give 1-(decan-2-yl) 15-(heptadecan-9-yl) 8-((2-oxaspiro[3.3]heptan-6-yl)amino)pentadecanedioate (18.20 mg, 52.9%) as a colorless oil. 1 H NMR (500MHz, methanol-d4, 27℃) δ ppm 0.92(9H,t),1.19-1.71(65H,m),1.93-2.08(2H,m),2.26-2.38(4H,m),2.50-2.61 (3H,m),3.17-3.29(1H,m),4.61(2H,s),4.74(2H,s),4.87-4.94(2H,m);C48H91NO5 m / z Calculated value 761.690 Actual value 762.7[M+H]+(LCMS).

[0192] Scheme 10 below illustrates the synthetic procedure for preparing Example 26.

[0193] Scheme 10: [ka] Reagents a) EDC.HCl, DIPEA, DMAP, DCM; b) NaBH(OAc)3, 1,2-DCE:NMP(4:1)

[0194] Example 26. Synthesis of Compound 26: 1-(heptadecan-9-yl) 15-(8-methylnonyl) 8-((2-oxaspiro[3.3]heptan-6-yl)amino)pentadecanedioate Step 1: 1-(heptadecan-9-yl) 15-(8-methylnonyl) 8-oxopentadecane dioate To a stirred solution of 15-(heptadecan-9-yloxy)-8,15-dioxopentadecanoic acid (0.03 g, 0.06 mmol), 8-methylnonan-1-ol (0.019 g, 0.12 mmol), DIPEA (0.031 mL, 0.18 mmol) and DMAP (1.397 mg, 0.01 mmol) in DCM (5 mL) at 0° C. under argon was added EDC (0.023 g, 0.12 mmol) in one portion. The resulting solution was stirred at room temperature for 16 h. The reaction mixture was concentrated to dryness under reduced pressure and redissolved in EtOAc (20 mL) and sodium bicarbonate (20 mL). The layers were separated and the aqueous layer was extracted with (EtOAc) (3×30 mL). The combined organic layers were washed successively with 5% citric acid (25 mL) and saturated aqueous NaCl (25 mL). The organic layer was dried over MgSO4, filtered and concentrated to dryness under reduced pressure to give the crude product. The resulting residue was purified by flash silica chromatography, elution gradient 0 to 60% EtOAc in hexanes. The product fractions were concentrated to dryness under reduced pressure to give 1-(heptadecan-9-yl) 15-(8-methylnonyl) 8-oxopentadecanedioate (0.028 g, 72.3%) as a colorless oil. 1 H NMR (500MHz, chloroform-d, 23℃) δ ppm 0.82-0.93(12H,m),1.10-1.19(2H,m),1.21-1.40(40H,m),1.46-1.68( 15H,m),2.25-2.34(4H,m),2.38(4H,t),4.06(2H,t),4.80-4.94(1H,m).

[0195] Compound 26: [ka] To a stirred solution of 2-oxaspiro[3.3]heptan-6-amine hydrochloride (0.019 g, 0.12 mmol) and 1-(heptadecan-9-yl)15-(8-methylnonyl)8-oxopentadecandioate (0.0275 g, 0.04 mmol) in DCE (4 mL) and NMP (1 mL) at 0° C. under argon was added sodium triacetoxyborohydride (0.026 g, 0.12 mmol) in one portion. The resulting solution was stirred at room temperature for 16 h. The reaction mixture was diluted with DCM (50 mL) and saturated sodium carbonate (50 mL). The layers were separated and the aqueous layer was extracted with (DCM) (3×25 mL). The organic layer was dried over MgSO4, filtered and concentrated to dryness under reduced pressure to give the crude product. The resulting residue was purified by flash silica chromatography, elution gradient 0 to 100% in DCM (20% MeOH and 1% NH4OH in DCM). The product fractions were concentrated to dryness under reduced pressure to give 1-(heptadecan-9-yl) 15-(8-methylnonyl) 8-((2-oxaspiro[3.3]heptan-6-yl)amino)pentadecanedioate (0.023 g, 72.3%) as a colourless oil. 1 H NMR (500MHz, chloroform-d, 27℃) δ ppm 0.84-0.93(12H,m),1.12-1.68(61H,m),1.79-1.91(2H,m),2.29(4H,m),2.38-2.47(1H,m),2.50 -2.59(2H,m),3.07-3.18(1H,m),4.06(2H,t),4.60(2H,s),4.71(2H,s),4.87(1H,m);C48H91NO5 m / z Calculated value 761.690 Actual value 762.9[M+H]+(LCMS).

[0196] Scheme 11 below illustrates the synthetic procedure for preparing Example 27.

[0197] Scheme 11: [ka] Reagents: a) EDC.HCl, DIPEA, DMAP, DCM; b) NaBH(OAc)3, 1,2-DCE:NMP(4:1)

[0198] Example 27. Synthesis of Compound 27: 1-(heptadecan-9-yl) 15-(3-heptyldecyl) 8-((2-oxaspiro[3.3]heptan-6-yl)amino)pentadecanedioate Step 1: 1-(heptadecan-9-yl) 15-(3-hexylnonyl) 8-oxopentadecane diate To a stirred mixture of 15-(heptadecan-9-yloxy)-8,15-dioxopentadecanoic acid (60 mg, 0.11 mmol), 3-heptyldecan-1-ol (44.0 mg, 0.17 mmol), DIPEA (0.082 mL, 0.47 mmol) and DMAP (2.79 mg, 0.02 mmol) in DCM (5 mL) at 0° C. under argon was added EDC (46.0 mg, 0.24 mmol) in one portion. The resulting mixture was stirred at room temperature for 16 h. The reaction mixture was diluted with sodium bicarbonate (25 mL) and DCM (25 mL). The layers were separated and the aqueous layer was extracted with (DCM) (4×25 mL). The combined organic layers were dried over MgSO4, filtered and concentrated to dryness under reduced pressure to give the crude product. The resulting residue was purified by flash silica chromatography, elution gradient 0 to 20% EtOAc in hexanes. The product fractions were concentrated to dryness under reduced pressure to give 1-(heptadecan-9-yl) 15-(3-heptyldecyl) 8-oxopentadecanedioate (60.0 mg, 68.8%) as a colorless oil. 1 H NMR (500MHz, chloroform-d, 21℃) δ ppm 0.89(12H,m),1.26(71H,m),2.24-2.32(4H,m),2.34-2.42(4H,t),4.03-4.13(2H,t),4.81-4.91(1H,m).

[0199] Compound 27: [ka] To a stirred solution of 2-oxaspiro[3.3]heptan-6-amine hydrochloride (0.013 g, 0.09 mmol) and 1-(heptadecan-9-yl) 15-(3-heptyldecyl) 8-oxopentadecandioate (0.0223 g, 0.03 mmol) in DCE (4 mL) and NMP (1 mL) at 0° C. under argon was added sodium triacetoxyborohydride (0.019 g, 0.09 mmol) in one portion. The resulting solution was stirred at room temperature for 16 h. The reaction mixture was diluted with DCM (50 mL) and saturated sodium carbonate (50 mL). The layers were separated and the aqueous layer was extracted with (DCM) (3×25 mL). The organic layer was dried over MgSO4, filtered and concentrated to dryness under reduced pressure to give the crude product. The resulting residue was purified by flash silica chromatography, elution gradient 0 to 100% in DCM (20% MeOH and 1% NH4OH in DCM). The product fractions were concentrated to dryness under reduced pressure to give 1-(heptadecan-9-yl) 15-(3-heptyldecyl) 8-((2-oxaspiro[3.3]heptan-6-yl)amino)pentadecanedioate (0.014 g, 55.7%) as a colourless oil. 1 H NMR (500MHz, chloroform-d, 27℃) δ ppm 0.89(12H,m),1.20-1.68(75H,m),1.81-1.93(2H,m),2.23-2.34(4H,m),2.40-2.49(1H,m),2.51-2.60 (2H,m),3.07-3.21(1H,m),4.05-4.12(2H,t),4.60(2H,s),4.71(2H,s),4.82-4.92(1H,m);C55H105NO5 m / z Calculated value 859.799 Actual value 861.0[M+H]+(LCMS).

[0200] Scheme 12 below illustrates the synthetic procedure for preparing Examples 28 and 29.

[0201] Scheme 12: [ka] Reagents: a, b) EDC.HCl, DIPEA, DMAP, DCM; c, d) NaBH(OAc)3,1,2-DCE:NMP(4:1)

[0202] Example 28. Synthesis of Compound 28: Bis(2-heptylnonyl) 8-((2-oxaspiro[3.3]heptan-6-yl)amino)pentadecanedioate Step 1: Bis(2-heptylnonyl) 8-oxopentadecane dioate and 15-((2-heptylnonyl)oxy)-8,15-dioxopentadecanoic acid To a stirred solution of 2-heptylnonan-1-ol (0.135 g, 0.56 mmol), 8-oxopentadecanedioic acid (0.200 g, 0.70 mmol), DIPEA (0.378 mL, 2.17 mmol) and DMAP (0.017 g, 0.14 mmol) in DCM (5 mL) at 0° C. under argon was added EDC (0.201 g, 1.05 mmol) in one portion. The resulting solution was stirred at room temperature for 16 h. The reaction mixture was concentrated to dryness under reduced pressure and redissolved in EtOAc (20 mL) and sodium bicarbonate (20 mL). The layers were separated and the aqueous layer was extracted with (EtOAc) (3×30 mL). The combined organic layers were washed successively with 5% citric acid (25 mL) and saturated aqueous NaCl (25 mL). The organic layers were dried over MgSO4, filtered and concentrated to dryness under reduced pressure to give the crude product. The resulting residue was purified by flash silica chromatography, elution gradient 0 to 60% EtOAc in hexane. The product fractions were concentrated to dryness under reduced pressure to give bis(2-heptylnonyl) 8-oxopentadecane dioate (0.060 g, 11.72%) and 15-((2-heptylnonyl)oxy)-8,15-dioxopentadecane dioate (0.126 g, 35.4%) as colorless oils. Bis(2-heptylnonyl) 8-oxopentadecane dioate: 1 H NMR (500 MHz, chloroform-d, 22° C.) δ ppm 0.83-0.95 (12H, m), 1.19-1.37 (56H, m), 1.62 (10H, m), 2.25-2.34 (4H, t), 2.38 (4H, t), 3.97 (4H, d). 15-((2-heptylnonyl)oxy)-8,15-dioxopentadecanoic acid: 1 H NMR (500 MHz, chloroform-d, 22°C) δ ppm 0.83-0.95 (6H, m), 1.27 (32H, br m), 1.52-1.72 (10H, m), 2.24-2.45 (8H, m), 3.97 (2H, d).

[0203] Step 2: 1-(Decan-2-yl) 15-(2-heptylnonyl) 8-oxopentadecane dioate To a stirred solution of 15-((2-heptylnonyl)oxy)-8,15-dioxopentadecanoic acid (0.0703 g, 0.14 mmol)), decan-2-ol (0.040 mL, 0.21 mmol), N-ethyl-N-isopropylpropan-2-amine (0.072 mL, 0.41 mmol) and N,N-dimethylpyridin-4-amine (3.36 mg, 0.03 mmol) in DCM (10 mL) at 0° C. under argon was added EDC (0.055 g, 0.29 mmol) in one portion. The resulting solution was stirred at room temperature for 16 h. The reaction mixture was concentrated to dryness under reduced pressure and redissolved in EtOAc (20 mL) and sodium bicarbonate (20 mL). The layers were separated and the aqueous layer was extracted with (EtOAc) (3×30 mL). The organic layer was dried over MgSO4, filtered and concentrated to dryness under reduced pressure to give the crude product. The resulting residue was purified by flash silica chromatography, elution gradient 0 to 60% EtOAc in hexanes. The product fractions were concentrated to dryness under reduced pressure to give 1-(decan-2-yl) 15-(2-heptylnonyl) 8-oxopentadecandioate (0.041 g, 46.2%) as a colorless oil. 1 H NMR (500 MHz, chloroform-d, 21° C.) δ ppm 0.84-0.93 (9H, m), 1.15-1.70 (58H, m), 2.23-2.32 (4H, m), 2.38 (4H, t), 3.94-3.99 (2H, d), 4.81-4.95 (1H, m).

[0204] Compound 28: [ka] To a stirred solution of 2-oxaspiro[3.3]heptan-6-amine hydrochloride (0.013 g, 0.08 mmol) and bis(2-heptylnonyl)8-oxopentadecandioate (0.026 g, 0.04 mmol) in DCE (2 mL) and NMP (0.5 mL) at 0° C. under argon was added sodium triacetoxyborohydride (0.020 g, 0.10 mmol) in one portion. The resulting solution was stirred at room temperature for 16 h. The reaction mixture was diluted with DCM (50 mL) and saturated sodium carbonate (50 mL). The layers were separated and the aqueous layer was extracted with (DCM) (3×25 mL). The organic layer was dried over MgSO4, filtered and concentrated to dryness under reduced pressure to give the crude product. The resulting residue was purified by flash silica chromatography, elution gradient 0 to 100% in DCM (20% MeOH and 1% NH4OH in DCM). The product fractions were concentrated to dryness under reduced pressure to give bis(2-heptylnonyl) 8-((2-oxaspiro[3.3]heptan-6-yl)amino)pentadecanedioate (0.018 g, 60.8%) as a colorless oil. 1 H NMR (500MHz, chloroform-d, 21℃) δ ppm 0.84-0.93(12H,m),1.20-1.41(64H,m),1.56-1.68(6H,m),1.84-1.99(2H,m),2.30(4H,sm,2.41-2. 49(1H,m),2.52-2.59(2H,m),3.12-3.21(1H,m),3.97(4H,d),4.60(2H,s),4.71(2H,s);C53H101NO5 m / z Calculated value 831.768 Actual value 833.0[M+H]+(LCMS).

[0205] Example 29. Synthesis of Compound 29: 1-(decane-2-yl) 15-(2-heptylnonyl) 8-((2-oxaspiro[3.3]heptane-6-yl)amino)pentadecanedioate [ka] Compound 29 was prepared using 15-((2-heptylnonyl)oxy)-8,15-dioxopentadecanoic acid and 1-(decan-2-yl)15-(2-heptylnonyl)8-oxopentadecandioate according to the protocol described for compound 28 with the following additional steps: Sodium triacetoxyborohydride (0.036 g, 0.17 mmol) was added in one portion to a stirred solution of 2-oxaspiro[3.3]heptan-6-amine hydrochloride (0.023 g, 0.15 mmol) and 1-(decan-2-yl)15-(2-heptylnonyl)8-oxopentadecandioate (0.0414 g, 0.06 mmol) in DCE (2 mL) and NMP (0.5 mL) at 0° C. under argon. The resulting solution was stirred at room temperature for 16 h. The reaction mixture was diluted with DCM (50 mL) and saturated sodium carbonate (50 mL). The layers were separated and the aqueous layer was extracted with (DCM) (3x25 mL). The organic layer was dried over MgSO4, filtered and concentrated to dryness under reduced pressure to give the crude product. The resulting residue was purified by flash silica chromatography, elution gradient 0 to 100% in DCM (20% MeOH and 1% NH4OH in DCM). The product fractions were concentrated to dryness under reduced pressure to give 1-(decan-2-yl) 15-(2-heptylnonyl) 8-((2-oxaspiro[3.3]heptan-6-yl)amino)pentadecanedioate (0.028 g, 59.5%) as a colorless oil. 1 H NMR (500MHz, chloroform-d, 27℃) δ ppm 0.86-0.93(9H,m),1.16-1.74(62H,m),1.82-1.98(2H,m),2.24-2.33(4H,m),2.39-2.48(1H,m),2.51-2. 59(2H,m),3.10-3.22(1H,m),3.94-4.01(2H,m),4.60(2H,s),4.71(2H,s),4.85-4.94(1H,m);C47H89NO5 m / z Calculated value 747.674 Actual value 748.9[M+H]+(LCMS).

[0206] Scheme 13 below illustrates the synthetic procedure for preparing Example 30.

[0207] Scheme 13: [ka] Reagents: a) EDC.HCl, DIPEA, DMAP, DCM; b) NaBH(OAc)3, 1,2-DCE:NMP(4:1)

[0208] Example 30. Synthesis of Compound 30: 1-(2-heptylnonyl) 15-(8-methylnonyl) 8-((2-oxaspiro[3.3]heptan-6-yl)amino)pentadecanedioate Step 1: 1-(2-heptylnonyl)15-(8-methylnonyl)8-oxopentadecane diate To a stirred solution of 15-((2-heptylnonyl)oxy)-8,15-dioxopentadecanoic acid (0.060 g, 0.12 mmol)), 8-methylnonan-1-ol (0.039 g, 0.25 mmol), DIPEA (0.064 mL, 0.36 mmol) and DMAP (2.87 mg, 0.02 mmol) in DCM (5 mL) at 0° C. under argon was added EDC (0.047 g, 0.25 mmol) in one portion. The resulting solution was stirred at room temperature for 16 h. The reaction mixture was concentrated to dryness under reduced pressure and redissolved in EtOAc (20 mL) and sodium bicarbonate (20 mL). The layers were separated and the aqueous layer was extracted with (EtOAc) (3×30 mL). The organic layer was dried over MgSO4, filtered and concentrated to dryness under reduced pressure to give the crude product. The resulting residue was purified by flash silica chromatography, elution gradient 0 to 60% EtOAc in hexanes. The product fractions were concentrated to dryness under reduced pressure to give 1-(2-heptylnonyl)15-(8-methylnonyl)8-oxopentadecandioate (0.071 g, 93%) as a colorless oil. 1 H NMR (500MHz, chloroform-d, 27 1 H)δ ppm 0.87(12H,m),1.12-1.67(54H,m),2.28(4H,m),2.34-2.41(4H,m),3.96(2H,d),4.05(2H,t).

[0209] Compound 30: [ka] To a stirred solution of 2-oxaspiro[3.3]heptan-6-amine hydrochloride (0.049 g, 0.33 mmol) and 1-(2-heptylnonyl)15-(8-methylnonyl)8-oxopentadecandioate (0.071 g, 0.11 mmol) in DCE (4 mL) and NMP (1 mL) at 0° C. under argon was added sodium triacetoxyborohydride (0.069 g, 0.33 mmol) in one portion. The resulting solution was stirred at room temperature for 16 h. The reaction mixture was diluted with DCM (50 mL) and saturated sodium carbonate (50 mL). The layers were separated and the aqueous layer was extracted with (DCM) (3×25 mL). The organic layer was dried over MgSO4, filtered and concentrated to dryness under reduced pressure to give the crude product. The resulting residue was purified twice by flash silica chromatography, elution gradient 0 to 100% in DCM (20% MeOH and 1% NH4OH in DCM). The product fractions were concentrated to dryness under reduced pressure to give 1-(2-heptylnonyl) 15-(8-methylnonyl) 8-((2-oxaspiro[3.3]heptan-6-yl)amino)pentadecanedioate (0.033 g, 40.1%) as a colourless oil. 1 H NMR (500MHz, chloroform-d, 27℃) δ ppm 0.84-0.92(12H,m),1.12-1.69(58H,m),1.81-1.92(2H,m),2.24-2.34(4H,m),2.38-2.45(1H,m),2.50 -2.59(2H,m),3.07-3.20(1H,m),3.95-3.99(2H,m),4.06(2H,t),4.60(2H,s),4.71(2H,s);;C47H89NO5 m / z Calculated value 747.674 Actual value 748.8[M+H]+(LCMS).

[0210] Scheme 14 below illustrates the synthetic procedures for preparing Examples 31 and 32.

[0211] Scheme 14: [ka] Reagents: a, b) EDC.HCl, DIPEA, DMAP, DCM; c, d) NaBH(OAc)3,1,2-DCE:NMP(4:1)

[0212] Example 31. Synthesis of Compound 31: Di(heptadecan-9-yl) 9-((2-oxaspiro[3.3]heptan-6-yl)amino)heptadecanedioate Step 1: Di(heptadecan-9-yl) 9-oxoheptadecanedioate and 17-(heptadecan-9-yloxy)-9,17-dioxoheptadecanoic acid To a stirred solution of 9-heptadecanol (0.098 g, 0.38 mmol), 9-oxoheptadecanedioic acid (0.15 g, 0.48 mmol), DIPEA (0.258 mL, 1.48 mmol) and DMAP (0.012 g, 0.10 mmol) in DCM (5 mL) at 0° C. under argon was added EDC (0.137 g, 0.72 mmol) in one portion. The resulting solution was stirred at room temperature for 16 h. The reaction mixture was concentrated to dryness under reduced pressure and redissolved in EtOAc (20 mL) and sodium biocarbonate (20 mL). The layers were separated and the aqueous layer was extracted with EtOAc (3×30 mL). The combined organic layers were washed successively with 5% citric acid (25 mL) and saturated aqueous NaCl (25 mL). The organic layer was dried over MgSO4, filtered and concentrated to dryness under reduced pressure to give the crude product. The resulting residue was purified by flash silica chromatography, elution gradient 0 to 60% EtOAc in hexanes. The product fractions were concentrated to dryness under reduced pressure to give di(heptadecan-9-yl) 9-oxoheptadecanedioate (0.047 g, 12.50%) and 17-(heptadecan-9-yloxy)-9,17-dioxoheptadecanoic acid (0.105 g, 39.7%) as colorless oils. Di(heptadecan-9-yl) 9-oxoheptadecanedioate: 1 H NMR (500 MHz, chloroform-d, 22° C.) δ ppm 0.88 (12H,t), 1.18-1.68 (76H,m), 2.23-2.32 (4H,t), 2.35-2.42 (4H,t), 4.87 (2H,m). 17-(heptadecan-9-yloxy)-9,17-dioxoheptadecanoic acid: 1 H NMR (500 MHz, chloroform-d, 22°C) δ ppm 0.83-0.95 (6H,m), 1.18-1.39 (36H,m), 1.45-1.68 (12H,m), 2.38 (8H,m), 4.77-4.96 (1H,m).

[0213] Step 2: 1-(Decan-2-yl) 15-(2-heptylnonyl) 8-oxopentadecane dioate To a stirred solution of 15-((2-heptylnonyl)oxy)-8,15-dioxopentadecanoic acid (0.0703 g, 0.14 mmol)), decan-2-ol (0.040 mL, 0.21 mmol), N-ethyl-N-isopropylpropan-2-amine (0.072 mL, 0.41 mmol) and N,N-dimethylpyridin-4-amine (3.36 mg, 0.03 mmol) in DCM (10 mL) at 0° C. under argon was added EDC (0.055 g, 0.29 mmol) in one portion. The resulting solution was stirred at room temperature for 16 h. The reaction mixture was concentrated to dryness under reduced pressure and redissolved in EtOAc (20 mL) and sodium bicarbonate (20 mL). The layers were separated and the aqueous layer was extracted with (EtOAc) (3×30 mL). The organic layer was dried over MgSO4, filtered and concentrated to dryness under reduced pressure to give the crude product. The resulting residue was purified by flash silica chromatography, elution gradient 0 to 60% EtOAc in hexanes. The product fractions were concentrated to dryness under reduced pressure to give 1-(decan-2-yl) 15-(2-heptylnonyl) 8-oxopentadecandioate (0.041 g, 46.2%) as a colorless oil. 1 H NMR (500 MHz, chloroform-d, 21° C.) δ ppm 0.84-0.93 (9H, m), 1.15-1.70 (58H, m), 2.23-2.32 (4H, m), 2.38 (4H, t), 3.94-3.99 (2H, d), 4.81-4.95 (1H, m).

[0214] Compound 31: [ka] To a stirred solution of 2-oxaspiro[3.3]heptan-6-amine hydrochloride (0.027 g, 0.18 mmol) and di(heptadecan-9-yl) 9-((2-oxaspiro[3.3]heptan-6-yl)amino)heptadecanedioate (0.026 g, 48.1%) in DCE (4 mL) and NMP (1 mL) at 0° C. under argon was added sodium triacetoxyborohydride (0.038 g, 0.18 mmol) in one portion. The resulting solution was stirred at room temperature for 16 h. The reaction mixture was diluted with DCM (50 mL) and saturated sodium carbonate (50 mL). The layers were separated and the aqueous layer was extracted with (DCM) (3×25 mL). The organic layer was dried over MgSO4, filtered and concentrated to dryness under reduced pressure to give the crude product. The resulting residue was purified by flash silica chromatography, elution gradient 0 to 100% in DCM (20% MeOH and 1% NH4OH in DCM). The product fractions were concentrated to dryness under reduced pressure to give di(heptadecan-9-yl) 9-((2-oxaspiro[3.3]heptan-6-yl)amino)heptadecanedioate (0.026 g, 48.1%) as a colorless oil. 1 H NMR (500MHz, methanol-d4, 22℃) δ ppm 0.91(12H,s),1.20-1.70(80H,m),1.97-2.07(2H,m),2.31(4H,s),2.51-2.61(3H ,m),3.03-3.20(1H,m),4.59(2H,s),4.69-4.76(2H,s),4.87(2H,m);C57H109NO5 m / z Calculated value 887.831 Actual value 889.0[M+H]+(LCMS).

[0215] Example 32. Compound 32: 1-(decane-2-yl) 17-(heptadecan-9-yl) 9-((2-oxaspiro[3.3]heptan-6-yl)amino)heptadecanedioate [ka] Compound 32 was prepared using 17-(heptadecan-9-yloxy)-9,17-dioxoheptadecanoic acid and 1-(decan-2-yl)15-(2-heptylnonyl)8-oxopentadecandioate according to the protocol described for compound 31 with the following additional steps: Sodium triacetoxyborohydride (0.040 g, 0.19 mmol) was added in one portion to a stirred solution of 2-oxaspiro[3.3]heptan-6-amine hydrochloride (0.028 g, 0.19 mmol) and 1-(decan-2-yl)17-(heptadecan-9-yl)9-oxoheptadecanedioate (0.044 g, 0.06 mmol) in DCE (4 mL) and NMP (1 mL) at 0° C. under argon. The resulting solution was stirred at room temperature for 16 h. The reaction mixture was diluted with DCM (50 mL) and saturated sodium carbonate (50 mL). The layers were separated and the aqueous layer was extracted with (DCM) (3x25 mL). The organic layer was dried over MgSO4, filtered and concentrated to dryness under reduced pressure to give the crude product. The resulting residue was purified by flash silica chromatography, elution gradient 0 to 100% in DCM (20% MeOH and 1% NH4OH in DCM). The product fractions were concentrated to dryness under reduced pressure to give 1-(decan-2-yl) 17-(heptadecan-9-yl) 9-((2-oxaspiro[3.3]heptan-6-yl)amino)heptadecanedioate (0.032 g, 63.4%) as a colorless oil. 1 H NMR (500MHz, methanol-d4, 27℃) δ ppm 0.92(9H,t),1.17-1.70(71H,m),1.95-2.07(2H,m),2.25-2.39(4H,m),2.50-2.62( 3H,m),3.20-3.29(1H,m),4.61(2H,s),4.74(2H,s),4.87-4.95(2H,m);C57H109NO5 m / z Calculated value 789.721 Actual value 790.7[M+H]+(LCMS).

[0216] Scheme 15 below illustrates the synthetic procedure for preparing Example 33.

[0217] Scheme 15: [ka] Reagents: a) EDC.HCl, DIPEA, DMAP, DCM; b) NaBH(OAc)3, 1,2-DCE:NMP(4:1)

[0218] Example 33. Synthesis of Compound 33: 1-(heptadecan-9-yl) 17-(2-heptylnonyl) 9-((2-oxaspiro[3.3]heptan-6-yl)amino)heptadecanedioate Step 1: 1-(heptadecan-9-yl) 17-(2-heptylnonyl) 9-oxoheptadecanedioate To a stirred solution of 17-(heptadecan-9-yloxy)-9,17-dioxoheptadecanoic acid (0.05 g, 0.09 mmol), 2-heptylnonan-1-ol (0.046 g, 0.19 mmol), DIPEA (0.049 mL, 0.28 mmol) and DMAP (2.210 mg, 0.02 mmol) in DCM (5 mL) at 0° C. under argon was added EDC (0.036 g, 0.19 mmol) in one portion. The resulting solution was stirred at room temperature for 16 h. The reaction mixture was concentrated to dryness under reduced pressure and redissolved in EtOAc (20 mL) and sodium biocarbonate (20 mL). The layers were separated and the aqueous layer was extracted with EtOAc (3×30 mL). The combined organic layers were washed successively with 5% citric acid (25 mL) and saturated aqueous NaCl (25 mL). The organic layer was dried over MgSO4, filtered, and concentrated to dryness under reduced pressure to give the crude product. The resulting residue was purified by flash silica chromatography, elution gradient 0 to 60% EtOAc in hexanes. The product fractions were concentrated to dryness under reduced pressure to give 1-(heptadecan-9-yl) 17-(2-heptylnonyl) 9-oxoheptadecanedioate (0.039 g, 55.8%) as a colorless oil. 1 H NMR (500MHz, chloroform-d, 22℃) δ ppm 0.89(12H,m),1.27(59H,m),1.46-1.68(14H,m),2.24-2.32(4H,m),2.34-2.41(4H,t),3.91-4.01(2H,d),4.80-4.93(1H,m).

[0219] Compound 33: [ka] To a stirred solution of 2-oxaspiro[3.3]heptan-6-amine hydrochloride (0.023 g, 0.15 mmol) and 1-(heptadecan-9-yl)17-(2-heptylnonyl)9-oxoheptadecanedioate (0.0392 g, 0.05 mmol) in DCE (4 mL) and NMP (1 mL) at 0° C. under argon was added sodium triacetoxyhydroborate (0.033 g, 0.16 mmol) in one portion. The resulting solution was stirred at room temperature for 16 h. The reaction mixture was diluted with DCM (50 mL) and saturated sodium carbonate (50 mL). The layers were separated and the aqueous layer was extracted with (DCM) (3×25 mL). The organic layer was dried over MgSO4, filtered and concentrated to dryness under reduced pressure to give the crude product. The resulting residue was purified by flash silica chromatography, elution gradient 0 to 100% in DCM (20% MeOH and 1% NH4OH in DCM). The product fractions were concentrated to dryness under reduced pressure to give 1-(heptadecan-9-yl) 17-(2-heptylnonyl) 9-((2-oxaspiro[3.3]heptan-6-yl)amino)heptadecanedioate (0.032 g, 72.3%) as a colourless oil. 1 H NMR (500MHz, chloroform-d, 22℃) δ ppm 0.89(12H,m),1.19-1.71(77H,m),1.80-1.90(2H,m),2.24-2.34(4H,m),2.37-2.46(1H,m),2.51-2. 59(2H,m),3.09-3.20(1H,m),3.92-4.02(2H,d),4.60(2H,s),4.71(2H,s),4.87(1H,m);C56H107NO5 m / z Calculated value 873.815 Actual value 875.0[M+H]+(LCMS).

[0220] Scheme 16 below illustrates the synthetic procedure for preparing Example 34.

[0221] Scheme 16: [ka] Reagents: a) EDC.HCl, DIPEA, DMAP, DCM; b) NaBH(OAc)3, 1,2-DCE:NMP(4:1)

[0222] Example 34. Synthesis of Compound 34: Bis(2-hexyloctyl) 9-((2-oxaspiro[3.3]heptan-6-yl)amino)heptadecanedioate Step 1: Bis(2-hexyloctyl) 9-oxoheptadecanedioate To a stirred solution of 9-oxoheptadecanedioic acid (0.100 g, 0.32 mmol), 2-hexyloctan-1-ol (0.150 g, 0.70 mmol), DMAP (7.77 mg, 0.06 mmol) and DIPEA (0.228 mL, 1.30 mmol) in MeOH (12 mL) at 25 °C under argon was added EDC (0.189 g, 0.99 mmol) in one portion. The resulting solution was stirred at room temperature for 24 h. The reaction mixture was diluted with DCM (50 mL) and water (50 mL). The layers were separated and the aqueous layer was extracted with (DCM) (3 x 50 mL). The organic layer was dried over MgSO4, filtered and evaporated to dryness to give the crude product. The resulting residue was purified by flash silica chromatography, elution gradient 0 to 40% EtOAc in hexanes. The product fractions were evaporated to dryness to give bis(2-hexyloctyl) 9-oxoheptadecandioate (0.195 g, 87%) as a colorless oil. 1 H NMR (500 MHz, chloroform-d, 27° C.) δ ppm 0.86 (12H, t), 1.25 (62H, m), 2.21-2.30 (4H, m), 2.35 (4H, s), 3.94 (4H, d).

[0223] Compound 34: [ka] To a stirred solution of 2-oxaspiro[3.3]heptan-6-amine hydrochloride (0.025 g, 0.17 mmol) and bis(2-hexyloctyl) 9-oxoheptadecanedioate (0.050 g, 0.07 mmol) in DCE (2 mL) and NMP (0.5 mL) at 0° C. under argon was added sodium triacetoxyborohydride (0.040 g, 0.19 mmol) in one portion. The resulting solution was stirred at room temperature for 16 h. The reaction mixture was diluted with DCM (50 mL) and saturated sodium carbonate (50 mL). The layers were separated and the aqueous layer was extracted with (DCM) (3×25 mL). The organic layer was dried over MgSO4, filtered and concentrated to dryness under reduced pressure to give the crude product. The resulting residue was purified by flash silica chromatography, elution gradient 0 to 100% in DCM (20% MeOH and 1% NH4OH in DCM). The product fractions were concentrated to dryness under reduced pressure to give bis(2-hexyloctyl) 9-((2-oxaspiro[3.3]heptan-6-yl)amino)heptadecandioate (0.034 g, 59.8%) as a colorless oil. 1 H NMR (500MHz, chloroform-d, 21℃) δ ppm 0.82-0.92(12H,m),1.17-1.38(60H,m),1.56-1.65(6H,m),1.79-1.88(2H,m),2.29(4H,t),2.38-2 .45(1H,m),2.46-2.59(2H,m),3.07-3.18(1H,m),3.96(4H,d),4.59(2H,s),4.70(2H,c);C51H97NO5 m / z Calculated value 803.737 Actual value 804.80[M+H]+(LCMS).

[0224] Scheme 17 below illustrates the synthetic procedures for preparing Examples 35-37.

[0225] Scheme 17: [ka] Reagents: a) EDC.HCl, DIPEA, DMAP, DCM; b) NaBH(OAc)3, 1,2-DCE:NMP(4:1)

[0226] Example 35. Synthesis of Compound 35: 1-(decane-2-yl) 17-(heptadecan-9-yl) 9-(((tetrahydro-2H-pyran-4-yl)methyl)amino)heptadecanedioate Step 1: 1-(Decan-2-yl) 17-(heptadecan-9-yl) 9-((2-oxaspiro[3.3]heptan-6-yl)amino)heptadecanedioate To a stirred mixture of 17-(heptadecan-9-yloxy)-9,17-dioxoheptadecanoic acid (160 mg, 0.29 mmol), decan-2-ol (0.083 mL, 0.43 mmol), DIPEA (0.207 mL, 1.19 mmol) and N,N-dimethylpyridin-4-amine (7.07 mg, 0.06 mmol) in DCM (5 mL) under argon at 0° C. was added 3-(((ethylimino)methylene)amino)-N,N-dimethylpropan-1-amine hydrochloride (117 mg, 0.61 mmol) in one portion. The resulting mixture was stirred at room temperature for 16 h. The reaction mixture was diluted with sodium bicarbonate (25 mL) and DCM (25 mL). The layers were separated and the aqueous layer was extracted with (DCM) (4×25 mL). The combined organic layers were dried over MgSO4, filtered and concentrated to dryness under reduced pressure to give the crude product. The resulting residue was purified by flash silica chromatography, elution gradient 0 to 40% EtOAc in hexanes. The product fractions were concentrated to dryness under reduced pressure to give 1-(decan-2-yl) 17-(heptadecan-9-yl) 9-oxoheptadecanedioate (91 mg, 45.4%) as a colorless oil. 1 H NMR (500 MHz, chloroform-d, 27° C.) δ ppm 0.89 (9H, t), 1.15-1.70 (65H, m), 2.27 (4H, m), 2.35-2.42 (4H, t), 4.80-4.96 (2H, m).

[0227] Compound 35: [ka] To a stirred solution of (tetrahydro-2H-pyran-4-yl)methanamine hydrochloride (0.024 g, 0.16 mmol) and 1-(decane-2-yl)17-(heptadecan-9-yl)9-oxoheptadecanedioate (0.04 g, 0.06 mmol) in DCE (2 mL) and NMP (0.5 mL) at 0° C. under argon was added sodium triacetoxyborohydride (0.038 g, 0.18 mmol) in one portion. The resulting solution was stirred at room temperature for 16 h. The reaction mixture was diluted with DCM (50 mL) and saturated sodium carbonate (50 mL). The layers were separated and the aqueous layer was extracted with (DCM) (3×25 mL). The organic layer was dried over MgSO4, filtered and concentrated to dryness under reduced pressure to give the crude product. The resulting residue was purified by flash silica chromatography, elution gradient 0 to 100% in DCM (20% MeOH and 1% NH4OH in DCM). The product fractions were concentrated to dryness under reduced pressure to give 1-(decane-2-yl) 17-(heptadecan-9-yl) 9-(((tetrahydro-2H-pyran-4-yl)methyl)amino)heptadecanedioate (0.034 g, 74.1%) as a colorless oil. 1 H NMR (500MHz, chloroform-d, 27℃) δ ppm 0.89(9H,t),1.17-1.79(74H,m),2.20-2.35(4H,m),2.40-2.58(3H,m),3.32-3.46(2H,t),3.92-4.02(2H,m),4.82-4.95(2H,m);C50H97NO5 m / z Calculated value 791.737 Actual value 792.7[M+H]+(LCMS).

[0228] Example 36. Compound 36: 1-(decane-2-yl) 17-(heptadecan-9-yl) 9-(((tetrahydrofuran-3-yl)methyl)amino)heptadecanedioate [ka] Compound 36 was prepared using 1-(decane-2-yl) 17-(heptadecan-9-yl) 9-oxoheptadecanedioate according to the protocol described for compound 35 with the following additional steps: Sodium triacetoxyborohydride (24.77 mg, 0.12 mmol) was added in one portion to a stirred solution of (tetrahydrofuran-3-yl)methanamine hydrochloride (14.29 mg, 0.10 mmol) and 1-(decane-2-yl) 17-(heptadecan-9-yl) 9-oxoheptadecanedioate (30 mg, 0.04 mmol) in DCE (2 mL) and NMP (0.5 mL) at 0° C. under argon. The resulting solution was stirred at room temperature for 16 h. The reaction mixture was diluted with DCM (50 mL) and saturated sodium carbonate (50 mL). The layers were separated and the aqueous layer was extracted with (DCM) (3x25 mL). The organic layer was dried over MgSO4, filtered and concentrated to dryness under reduced pressure to give the crude product. The resulting residue was purified by flash silica chromatography, elution gradient 0 to 100% in DCM (20% MeOH and 1% NH4OH in DCM). The product fractions were concentrated to dryness under reduced pressure to give 1-(decan-2-yl) 17-(heptadecan-9-yl) 9-(((tetrahydrofuran-3-yl)methyl)amino)heptadecanedioate (26.5 mg, 79%) as a colorless oil. 1 H NMR (500MHz, methanol-d4, 27℃) δ ppm 0.90(9H,t),1.17-1.67(70H,m),2.02-2.18(1H,m),2.22-2.35(4H,m),2.35-2.46(1H,m),2.51-2.59(1H,m ),2.60-2.67(2H,d),3.43-3.51(1H,m),3.68-3.78(1H,m),3.81-3.92(2H,m),4.85-4.93(2H,m);C49H95NO5 m / z Calculated value 777.721 Actual value 778.8[M+H]+(LCMS).

[0229] Example 37. Compound 37: 1-(decane-2-yl) 17-(heptadecan-9-yl) 9-((oxetan-3-ylmethyl)amino)heptadecanedioate [ka] Compound 37 was prepared using 1-(decane-2-yl) 17-(heptadecan-9-yl) 9-oxoheptadecanedioate according to the protocol described for compound 35 with the following additional steps: Sodium triacetoxyborohydride (24.77 mg, 0.12 mmol) was added in one portion to a stirred solution of oxetan-3-ylmethanamine hydrochloride (12.84 mg, 0.10 mmol) and 1-(decane-2-yl) 17-(heptadecan-9-yl) 9-oxoheptadecanedioate (30 mg, 0.04 mmol) in DCE (2 mL) and NMP (0.5 mL) at 0° C. under argon. The resulting solution was stirred at room temperature for 16 h. The reaction mixture was diluted with DCM (50 mL) and saturated sodium carbonate (50 mL). The layers were separated and the aqueous layer was extracted with (DCM) (3x25 mL). The organic layer was dried over MgSO4, filtered and concentrated to dryness under reduced pressure to give the crude product. The resulting residue was purified by flash silica chromatography, elution gradient 0 to 100% in DCM (20% MeOH and 1% NH4OH in DCM). The product fractions were concentrated to dryness under reduced pressure to give 1-(decan-2-yl) 17-(heptadecan-9-yl) 9-((oxetan-3-ylmethyl)amino)heptadecanedioate (19.80 mg, 59.9%) as a colorless oil. 1 H NMR (500MHz, methanol-d4, 27℃) δ ppm 0.90(9H,t),1.16-1.68(69H,m),2.31(4H,m),2.47-2.59(1H,m),2.91-2.97(2H,d),3 .07-3.16(1H,m),4.35-4.46(2H,t),4.75-4.82(2H,m),4.85-4.92(2H,m);C48H93NO5 m / z Calculated value 763.705 Actual value 764.7[M+H]+(LCMS).

[0230] Scheme 18 below illustrates the synthetic procedures for preparing Examples 38-41.

[0231] Scheme 18: [ka] Reagents: a) EDC.HCl, DIPEA, DMAP, DCM; b) NaBH(OAc)3, 1,2-DCE:NMP(4:1)

[0232] Example 38. Synthesis of Compound 38: 1-(heptadecan-9-yl) 17-(undecane-3-yl) 9-((2-oxaspiro[3.3]heptan-6-yl)amino)heptadecanedioate Step 1: 1-(heptadecan-9-yl) 17-(undecane-3-yl) 9-oxoheptadecanedioate To a stirred mixture of 17-(heptadecan-9-yloxy)-9,17-dioxoheptadecanoic acid (0.2 g, 0.36 mmol), undecan-3-ol (0.093 g, 0.54 mmol), DIPEA (0.195 mL, 1.12 mmol) and DMAP (8.84 mg, 0.07 mmol) in DCM (5 mL) at 0° C. under argon was added EDC (0.146 g, 0.76 mmol) in one portion. The resulting mixture was stirred at room temperature for 16 h. The reaction mixture was diluted with 10% citric acid (25 mL) and DCM (25 mL). The layers were separated and the aqueous layer was extracted with (DCM) (4×25 mL). The combined organic layers were dried over MgSO4, filtered and concentrated to dryness under reduced pressure to give the crude product. The resulting residue was purified by flash silica chromatography, elution gradient 0 to 40% EtOAc in hexanes. The product fractions were concentrated to dryness under reduced pressure to give 1-(heptadecan-9-yl) 17-(undecan-3-yl) 9-oxoheptadecanedioate (0.180 g, 70.4%) as a colorless oil. 1 H NMR (500 MHz, chloroform-d, 27° C.) δ ppm 0.89 (9H, t), 1.16-1.71 (67H, m), 2.23-2.30 (4H, m), 2.38 (4H, t), 4.82-4.95 (2H, m).

[0233] Compound 38: [ka] To a stirred solution of 2-oxaspiro[3.3]heptan-6-amine hydrochloride (0.023 g, 0.15 mmol) and 1-(heptadecan-9-yl)17-(undecan-3-yl)9-oxoheptadecanedioate (0.04 g, 0.06 mmol) in DCE (2 mL) and NMP (0.5 mL) at 0° C. under argon was added sodium triacetoxyborohydride (0.037 g, 0.18 mmol) in one portion. The resulting solution was stirred at room temperature for 16 h. The reaction mixture was diluted with DCM (50 mL) and saturated sodium carbonate (50 mL). The layers were separated and the aqueous layer was extracted with (DCM) (3×25 mL). The organic layer was dried over MgSO4, filtered and concentrated to dryness under reduced pressure to give the crude product. The resulting residue was purified by flash silica chromatography, elution gradient 0 to 100% in DCM (20% MeOH and 1% NH4OH in DCM). The product fractions were concentrated to dryness under reduced pressure to give 1-(heptadecan-9-yl) 17-(undecan-3-yl) 9-((2-oxaspiro[3.3]heptan-6-yl)amino)heptadecanedioate (0.027 g, 59.3%) as a colourless oil. 1 H NMR (500MHz, methanol-d4, 27℃) δ ppm 0.87-0.96(12H,m),1.26-1.70(68H,m),1.95-2.06(2H,m),2.29-2.37(4H,m),2.49-2.60(3H,m),3 .18-3.29(1H,m),4.58-4.63(2H,s),4.73(2H,s),4.80-4.84(1H,m),4.87-4.94(1H,m);C51H97NO5 m / z Calculated value 803.737 Actual value 804.72[M+H]+(LCMS).

[0234] Example 39. Synthesis of Compound 39: 1-(decane-2-yl) 17-(heptadecan-9-yl) 9-((oxetan-3-ylmethyl)amino)heptadecanedioate [ka] Compound 39 was prepared using 1-(heptadecan-9-yl) 17-(undecan-3-yl) 9-oxoheptadecanedioate according to the protocol described for compound 38 with the following additional steps: Sodium triacetoxyborohydride (0.028 g, 0.13 mmol) was added in one portion to a stirred solution of (tetrahydro-2H-pyran-4-yl)methanamine hydrochloride (0.017 g, 0.11 mmol) and 1-(heptadecan-9-yl) 17-(undecan-3-yl) 9-oxoheptadecanedioate (0.03 g, 0.04 mmol) in DCE (2 mL) and NMP (0.5 mL) at 0° C. under argon. The resulting solution was stirred at room temperature for 16 h. The reaction mixture was diluted with DCM (50 mL) and saturated sodium carbonate (50 mL). The layers were separated and the aqueous layer was extracted with (DCM) (3x25 mL). The organic layer was dried over MgSO4, filtered and concentrated to dryness under reduced pressure to give the crude product. The resulting residue was purified by flash silica chromatography, elution gradient 0 to 100% in DCM (20% MeOH and 1% NH4OH in DCM). The product fractions were concentrated to dryness under reduced pressure to give 1-(heptadecan-9-yl) 17-(undecan-3-yl) 9-(((tetrahydro-2H-pyran-4-yl)methyl)amino)heptadecanedioate (0.023 g, 67.8%) as a colorless oil. 1 H NMR (500 MHz, methanol-d4, 27 °C) δ ppm 0.84-0.97 (12H, m), 1.22-1.83 (73H, m), 2.26-2.37 (4H, m), 2.55-2.60 (2H, d), 2.62-2.71 (1H, m), 3.36-3.48 (2H, t), 3.90-3.98 (2H, m), 4.77-4.82 (1H, m), 4.85-4.93 (1H, m); C51H99NO5 m / z calculated 805.752 found 806.8 [M+H]+ (LCMS).

[0235] Example 40. Synthesis of Compound 40: 1-(heptadecan-9-yl) 17-(undecan-3-yl) 9-(((tetrahydrofuran-3-yl)methyl)amino)heptadecanedioate [ka] Compound 40 was prepared using 1-(heptadecan-9-yl) 17-(undecan-3-yl) 9-oxoheptadecanedioate according to the protocol described for compound 38 with the following additional steps: Sodium triacetoxyborohydride (32.4 mg, 0.15 mmol) was added in one portion to a stirred solution of (tetrahydrofuran-3-yl)methanamine hydrochloride (18.68 mg, 0.14 mmol) and 1-(heptadecan-9-yl) 17-(undecan-3-yl) 9-oxoheptadecanedioate (40 mg, 0.06 mmol) in DCE (2 mL) and NMP (0.5 mL) at 0° C. under argon. The resulting solution was stirred at room temperature for 16 h. The reaction mixture was diluted with DCM (50 mL) and saturated sodium carbonate (50 mL). The layers were separated and the aqueous layer was extracted with (DCM) (3x25 mL). The organic layer was dried over MgSO4, filtered and concentrated to dryness under reduced pressure to give the crude product. The resulting residue was purified by flash silica chromatography, elution gradient 0 to 100% in DCM (20% MeOH and 1% NH4OH in DCM). The product fractions were concentrated to dryness under reduced pressure to give 1-(heptadecan-9-yl) 17-(undecan-3-yl) 9-(((tetrahydrofuran-3-yl)methyl)amino)heptadecanedioate (28.7 mg, 64.0%) as a colorless oil. 1 H NMR (500MHz, methanol-d4, 27℃) δ ppm 0.84-0.97(12H,m),1.22-1.69(69H,m),2.05-2.16(1H,m),2.31(4H,br d),2.42(1H,s),2.59-2.65(1H,m),2.65-2.72(2H,d),3.44-3.52(1H,m),3.69- 3.77(1H,m),3.81-3.92(2H,m),4.74-4.82(1H,m),4.86-4.93(1H,m);C50H97NO5 m / z Calculated value 791.737 Actual value 792.8[M+H]+(LCMS).

[0236] Example 41. Synthesis of Compound 41: 1-(heptadecan-9-yl) 17-(undecane-3-yl) 9-((oxetan-3-ylmethyl)amino)heptadecanedioate [ka] Compound 41 was prepared using 1-(heptadecan-9-yl) 17-(undecan-3-yl) 9-oxoheptadecanedioate according to the protocol described for compound 38 with the following additional steps: Sodium triacetoxyborohydride (24.28 mg, 0.11 mmol) was added in one portion to a stirred solution of oxetan-3-ylmethanamine hydrochloride (12.58 mg, 0.10 mmol) and 1-(heptadecan-9-yl) 17-(undecan-3-yl) 9-oxoheptadecanedioate (30 mg, 0.04 mmol) in DCE (2 mL) and NMP (0.5 mL) at 0° C. under argon. The resulting solution was stirred at room temperature for 16 h. The reaction mixture was diluted with DCM (50 mL) and saturated sodium carbonate (50 mL). The layers were separated and the aqueous layer was extracted with (DCM) (3x25 mL). The organic layer was dried over MgSO4, filtered and concentrated to dryness under reduced pressure to give the crude product. The resulting residue was purified by flash silica chromatography, elution gradient 0 to 100% in DCM (20% MeOH and 1% NH4OH in DCM). The product fractions were concentrated to dryness under reduced pressure to give 1-(heptadecan-9-yl) 17-(undecan-3-yl) 9-((oxetan-3-ylmethyl)amino)heptadecanedioate (21.00 mg, 63.6%) as a colorless oil. 1 H NMR (500 MHz, methanol-d4, 27 °C) δ ppm 0 0.91 (12H, m), 1.20-1.71 (68H, m), 2.24-2.38 (4H, m), 2.46-2.57 (1H, m), 2.89-2.99 (2H, d), 3.06-3.17 (1H, m), 4.36-4.44 (2H, t), 4.77-4.82 (3H, m), 4.86-4.92 (1H, m); C49H95NO5 m / z calculated 777.721 found 778.6 [M+H]+ (LCMS).

[0237] Scheme 19 below illustrates the synthetic procedures for preparing Examples 42 and 43.

[0238] Scheme 19: [ka] Reagents: a) EDC.HCl, DIPEA, DMAP, DCM; b) NaBH(OAc)3, 1,2-DCE:NMP(4:1)

[0239] Example 42. Synthesis of Compound 42: 1-(heptadecan-9-yl) 17-(3-methylnonyl) 9-((2-oxaspiro[3.3]heptan-6-yl)amino)heptadecanedioate Step 1: 1-(heptadecan-9-yl) 17-(3-methylnonyl) 9-oxoheptadecanedioate To a stirred mixture of 17-(heptadecan-9-yloxy)-9,17-dioxoheptadecanoic acid (80 mg, 0.14 mmol), 3-methylnonan-1-ol (0.047 mL, 0.22 mmol), DIPEA (0.104 mL, 0.59 mmol) and DMAP (3.54 mg, 0.03 mmol) in DCM (5 mL) at 0° C. under argon was added EDC (58.3 mg, 0.30 mmol) in one portion. The resulting mixture was stirred at room temperature for 16 h. The reaction mixture was diluted with DCM (25 mL) and 10% citric acid (25 mL). The layers were separated and the aqueous layer was extracted with (DCM) (3×25 mL). The combined organic layers were dried over MgSO4, filtered and concentrated to dryness under reduced pressure to give the crude product. The resulting residue was purified by flash silica chromatography, elution gradient 0 to 60% EtOAc in hexanes. The product fractions were concentrated to dryness under reduced pressure to give 1-(heptadecan-9-yl) 17-(3-methylnonyl) 9-oxoheptadecanedioate (61.9 mg, 61.7%) as a white solid. 1 H NMR (500MHz, chloroform-d, 27℃) δ ppm 0.82-0.97(12H,m),1.10-1.70(61H,m),2.25-2.31(4H,m),2.38(4H,t),4.02-4.18(2H,m),4.80-4.93(1H,m).

[0240] Compound 42: [ka] To a stirred solution of 2-oxaspiro[3.3]heptan-6-amine hydrochloride (15.54 mg, 0.10 mmol) and 1-(heptadecan-9-yl)17-(3-methylnonyl)9-oxoheptadecanedioate (30 mg, 0.04 mmol) in DCE (2 mL) and NMP (0.5 mL) at 0° C. under argon was added sodium triacetoxyborohydride (24.77 mg, 0.12 mmol) in one portion. The resulting solution was stirred at room temperature for 16 h. The reaction mixture was diluted with DCM (50 mL) and saturated sodium carbonate (50 mL). The layers were separated and the aqueous layer was extracted with (DCM) (3×25 mL). The organic layer was dried over MgSO4, filtered and concentrated to dryness under reduced pressure to give the crude product. The resulting residue was purified by flash silica chromatography, elution gradient 0 to 100% in DCM (20% MeOH and 1% NH4OH in DCM). The product fractions were concentrated to dryness under reduced pressure to give 1-(heptadecan-9-yl) 17-(3-methylnonyl) 9-((2-oxaspiro[3.3]heptan-6-yl)amino)heptadecanedioate (16.90 mg, 49.4%) as a colorless oil. 1 H NMR (500MHz, methanol-d4, 27℃) δ ppm 0.85-0.98(12H,m),1.13-1.73(65H,m),1.96-2.08(2H,m),2.31(4H,t),2.48-2.59(3H,m),3 .18-3.27(1H,m),4.04-4.19(2H,m),4.59(2H,s),4.72(2H,s),4.85-4.92(1H,m);C50H95NO5 m / z Calculated value 789.721 Actual value 790.7[M+H]+(LCMS).

[0241] Example 43. Synthesis of Compound 43: 1-(heptadecan-9-yl) 17-(3-methylnonyl) 9-(((tetrahydro-2H-pyran-4-yl)methyl)amino)heptadecanedioate [ka] Compound 43 was prepared using 1-(heptadecan-9-yl) 17-(3-methylnonyl) 9-oxoheptadecanedioate according to the protocol described for compound 42 with the following additional steps: Sodium triacetoxyborohydride (24.77 mg, 0.12 mmol) was added in one portion to a stirred solution of (tetrahydro-2H-pyran-4-yl)methanamine hydrochloride (15.75 mg, 0.10 mmol) and 1-(heptadecan-9-yl) 17-(3-methylnonyl) 9-oxoheptadecanedioate (30 mg, 0.04 mmol) in DCE (2 mL) and NMP (0.5 mL) at 0° C. under argon. The resulting solution was stirred at room temperature for 16 h. The reaction mixture was diluted with DCM (50 mL) and saturated sodium carbonate (50 mL). The layers were separated and the aqueous layer was extracted with (DCM) (3x25 mL). The organic layer was dried over MgSO4, filtered and concentrated to dryness under reduced pressure to give the crude product. The resulting residue was purified by flash silica chromatography, elution gradient 0 to 100% in DCM (20% MeOH and 1% NH4OH in DCM). The product fractions were concentrated to dryness under reduced pressure to give 1-(heptadecan-9-yl) 17-(3-methylnonyl) 9-(((tetrahydro-2H-pyran-4-yl)methyl)amino) heptadecanedioate (9.30 mg, 27.1%) as a colorless oil. 1 H NMR (500MHz, methanol-d4, 27℃) δ ppm 0.85-0.97(12H,m),1.14-1.84(70H,m),2.31(4H,s),2.56-2.65(2H,m),2.63-2.76(1H,m ),3.36-3.49(2H,m),3.88-4.00(2H,m),4.04-4.19(2H,m),4.85-4.92(1H,m);C50H97NO5 m / z Calculated value 791.737 Actual value 792.7[M+H]+(LCMS).

[0242] Scheme 20 below illustrates the synthetic procedure for preparing Example 44.

[0243] Scheme 20: [ka] Reagents: a) EDC.HCl, DIPEA, DMAP, DCM; b) NaBH(OAc)3, 1,2-DCE:NMP(4:1)

[0244] Example 44. Synthesis of Compound 44: 1-(3-ethylnonyl) 17-(heptadecan-9-yl) 9-((2-oxaspiro[3.3]heptan-6-yl)amino)heptadecanedioate Step 1: 1-(3-ethylnonyl) 17-(heptadecan-9-yl) 9-oxoheptadecanedioate To a stirred mixture of 17-((3-ethylnonyl)oxy)-9,17-dioxoheptadecanoic acid (0.067 g, 0.14 mmol), heptadecan-9-ol (0.055 g, 0.21 mmol), DIPEA (0.102 mL, 0.59 mmol) and DMAP (3.49 mg, 0.03 mmol) in DCM (5 mL) at 0° C. under argon was added EDC (0.058 g, 0.30 mmol) in one portion. The resulting mixture was stirred at room temperature for 16 h. The reaction mixture was diluted with sodium bicarbonate (25 mL) and DCM (25 mL). The layers were separated and the aqueous layer was extracted with (DCM) (4×25 mL). The combined organic layers were dried over MgSO4, filtered and concentrated to dryness under reduced pressure to give the crude product. The resulting residue was purified by flash silica chromatography, elution gradient 0 to 40% EtOAc in hexanes. The product fractions were concentrated to dryness under reduced pressure to give 1-(3-ethylnonyl) 17-(heptadecan-9-yl) 9-oxoheptadecanedioate (0.040 g, 39.6%) as a colorless oil. 1 H NMR (500 MHz, chloroform-d, 27° C.) δ ppm 0.81-0.92 (12H, m), 1.17-1.68 (63H, m), 2.21-2.31 (4H, m), 2.34-2.41 (4H, t), 3.98-4.14 (2H, t), 4.79-4.91 (1H, m).

[0245] Compound 44: [ka] To a stirred solution of 1-(3-ethylnonyl) 17-(heptadecan-9-yl) 9-oxoheptadecanedioate (0.040 g, 0.06 mmol) and 1-(3-ethylnonyl) 17-(heptadecan-9-yl) 9-oxoheptadecanedioate (0.040 g, 0.06 mmol) in DCE (2 mL) and NMP (0.5 mL) at 0° C. under argon was added sodium triacetoxyborohydride (0.032 g, 0.15 mmol) in one portion. The resulting solution was stirred at room temperature for 16 h. The reaction mixture was diluted with DCM (50 mL) and saturated sodium carbonate (50 mL). The layers were separated and the aqueous layer was extracted with (DCM) (3×25 mL). The organic layer was dried over MgSO4, filtered and concentrated to dryness under reduced pressure to give the crude product. The resulting residue was purified by flash silica chromatography, elution gradient 0 to 100% in DCM (20% MeOH and 1% NH4OH in DCM). The product fractions were concentrated to dryness under reduced pressure to give 1-(3-ethylnonyl) 17-(heptadecan-9-yl) 9-((2-oxaspiro[3.3]heptan-6-yl)amino)heptadecanedioate (0.029 g, 64.4%) as a colourless oil. 1 H NMR (500MHz, methanol-d4, 27℃) δ ppm 0.91(12H,m),1.22-1.69(67H,m),1.96-2.05(2H,m),2.31(4H,t),2.49-2.59(3H,m),3.1 7-3.27(1H,m),4.07-4.15(2H,t),4.59(2H,s),4.72(2H,s),4.84-4.91(1H,m);C50H95NO5 m / z Calculated value 803.737 Actual value 804.6[M+H]+(LCMS).

[0246] Example 45. Synthesis of Compound 45: Bis(3-pentyloctyl) 7-((tetrahydro-2H-pyran-4-yl)amino) tridecane dioate [ka] Compound 45 was prepared using tetraethyl 6-oxoundecane-1,5,7,11-tetracarboxylate, 7-oxotridecanedioic acid and bis(3-pentyloctyl)7-oxotridecanedioate according to the protocol described for compound 13, with the following additional steps: Sodium triacetoxyhydroborate (44.5 mg, 0.21 mmol) was added in one portion to a stirred solution of bis(3-pentyloctyl)7-oxotridecanedioate (54.5 mg, 0.09 mmol), tetrahydro-2H-pyran-4-amine (19.02 μl, 0.18 mmol) and acetic acid (262 μl, 0.26 mmol) in DCM (2 mL) and NMP (0.5 mL) at 25° C. under argon. The resulting suspension was stirred at 25° C. for 40 h. The reaction mixture was diluted with DCM (15 mL), water (5 mL) and saturated Na2CO3 (10 mL). The layers were separated and the aqueous layer was extracted with DCM (3x15 mL). The combined organic layers were dried over MgSO4, filtered and concentrated to dryness under reduced pressure to give the crude product. The resulting residue was purified by flash silica chromatography, elution gradient 0 to 50% 20% MeOH / DCM (with 1% NH4OH) in DCM. The product fractions were concentrated to dryness under reduced pressure to give bis(3-pentyloctyl) 7-((tetrahydro-2H-pyran-4-yl)amino) tridecanedioate (5.10 mg, 8.23%) as a colorless oil. 1 H NMR(500MHz,methanol-d4)δ ppm 0.9(s,12H)1.3(br s,48H)1.6-1.7(m,8H)1.8-1.9(m,2H)2.3(t,J=7.3Hz,4H)2.7-2.8(m,1H)2.9-3.0(m,1H)3.4(br d,J=1.7Hz,2H)3.9-4.0(m,2H)4.1(t,J=6.8Hz,4H);C 44 H 85 NO5 m / z Calculated value 707.643 Measured value 708.7 [M+H] + (LCMS).

[0247] Scheme 21 below illustrates the synthetic procedure for preparing Example 46. In step w below, when the respective amine was utilized as the free base, AcOH was used as an additive in the reductive amination.

[0248] Scheme 21: [ka] Reagents: v) EDC.HCl, DIPEA, DMAP, DCM; w) NaBH(OAc)3, AcOH, 1,2-DCE:NMP (4:1)

[0249] Example 46. Synthesis of Compound 46: 1-(heptadecan-9-yl) 17-(octan-3-yl) 9-((2-oxaspiro[3.3]heptan-6-yl)amino)heptadecanedioate Step v): To a stirred solution of 17-(heptadecan-9-yloxy)-9,17-dioxoheptadecanoic acid (70.7 mg, 0.13 mmol), N-ethyl-N-isopropylpropan-2-amine (0.080 mL, 0.46 mmol), N,N-dimethylpyridin-4-amine (2.343 mg, 0.02 mmol) and octan-3-ol (0.043 mL, 0.27 mmol) in DCM (3 mL) at 0° C. under argon was added 3-(((ethylimino)methylene)amino)-N,N-dimethylpropan-1-amine hydrochloride (52.0 mg, 0.27 mmol) in one portion. The resulting solution was stirred at 25° C. for 16 h. The reaction mixture was diluted with EtOAc (20 mL), water (5 mL) and 5% citric acid solution (15 mL). The layers were separated and the aqueous layer was extracted with EtOAc (3x20 mL). The combined organic layers were washed with saturated aqueous NaCl (20 mL). The organic layers were dried over MgSO4, filtered and concentrated to dryness under reduced pressure to give the crude product. The resulting residue was purified by flash silica chromatography, elution gradient 0 to 35% EtOAc in hexanes. The product fractions were concentrated to dryness under reduced pressure to give 1-(heptadecan-9-yl) 17-(octan-3-yl) 9-oxoheptadecanedioate (52.6 mg, 61.8%) as a colorless oil. 1 H NMR (500 MHz, chloroform-d) 0.84-0.91 (m, 12H), 1.21-1.33 (m, 42H), 1.46-1.64 (m, 16H), 2.27 (td, J = 7.5, 5.3 Hz, 4H), 2.37 (t, J = 7.4 Hz, 4H), 4.78-4.89 (m, 2H).

[0250] Step W): [ka] (After 10 min) sodium triacetoxyhydroborate (50.3 mg, 0.24 mmol) was added in one portion to a stirred solution of 1-(heptadecan-9-yl) 17-(octan-3-yl) 9-oxoheptadecanedioate (52.6 mg, 0.08 mmol) and 2-oxaspiro[3.3]heptan-6-aminium chloride (34.3 mg, 0.23 mmol) in 1,2-DCE (2 mL) and NMP (0.5 mL) under argon. The resulting solution was stirred at 25° C. for 18 h. The reaction mixture was diluted with DCM (15 mL), water (5 mL) and saturated Na2CO3 (5 mL). The layers were separated and the aqueous layer was extracted with DCM (3×15 mL). The combined organic phase was dried over MgSO4, filtered and evaporated to dryness to give the crude product. The resulting residue was purified by flash silica chromatography, elution gradient 0 to 40% 20% MeOH in DCM (with 1% NH4OH) in DCM. The product fractions were concentrated to dryness under reduced pressure to give 1-(heptadecan-9-yl) 17-(octan-3-yl) 9-((2-oxaspiro[3.3]heptan-6-yl)amino)heptadecanedioate (25.9 mg, 43.0%) as a colorless oil. 1 H NMR (400MHz, methanol-d4)0.88-0.95(m,12H),1.26-1.39(m,50H),1.50-1.68(m,12H),1.94-2.02(m,2H),2.33(td,J=7.2 C 48 H 91 NO5 m / z Calculated value 761.690 Measured value 762.6 [M+H] + (LCMS).

[0251] Scheme 22 below illustrates the synthetic procedure for preparing Example 47. In step y below, when the respective amine was utilized as the free base, AcOH was used as an additive in the reductive amination.

[0252] Scheme 22: [ka] Reagents: x) EDC.HCl, DIPEA, DMAP, DCM; y) NaBH(OAc)3, AcOH, 1,2-DCE:NMP(4:1)

[0253] Example 47. Synthesis of compound 47: 1-(heptadecan-9-yl) 17-(heptan-3-yl) 9-(((tetrahydrofuran-3-yl)methyl)amino)heptadecanedioate Step x): To a stirred solution of 17-(heptadecan-9-yloxy)-9,17-dioxoheptadecanoic acid (63.5 mg, 0.11 mmol), N-ethyl-N-isopropylpropan-2-amine (0.072 mL, 0.41 mmol), N,N-dimethylpyridin-4-amine (2.105 mg, 0.02 mmol) and heptan-3-ol (0.038 mL, 0.26 mmol) in DCM (3 mL) at 0° C. under argon was added 3-(((ethylimino)methylene)amino)-N,N-dimethylpropan-1-amine hydrochloride (52.8 mg, 0.28 mmol) in one portion. The resulting solution was stirred at 25° C. for 16 h. The reaction mixture was diluted with EtOAc (20 mL), water (5 mL) and 5% citric acid solution (15 mL). The layers were separated and the aqueous layer was extracted with EtOAc (3x20 mL). The combined organic layers were washed with saturated aqueous NaCl (20 mL). The organic layers were dried over MgSO4, filtered and concentrated to dryness under reduced pressure to give the crude product. The resulting residue was purified by flash silica chromatography, elution gradient 0 to 35% EtOAc in hexanes. The product fractions were concentrated to dryness under reduced pressure to give 1-(heptadecan-9-yl) 17-(heptan-3-yl) 9-oxoheptadecanedioate (37.0 mg, 49.5%) as a colorless oil. 1 H NMR(500MHz,chloroform-d)δ ppm 0.8-0.9(m,12H)1.3(br s,40H)1.5-1.6(m,16H)2.2-2.3(m,4H)2.4(t,J=7.4Hz,4H)4.8-4.9(m,2H).

[0254] Step y): [ka] (After 10 min) Sodium triacetoxyhydroborate (33.7 mg, 0.16 mmol) was added in one portion to a stirred solution of 1-(heptadecan-9-yl) 17-(heptan-3-yl) 9-oxoheptadecanedioate (37 mg, 0.06 mmol) and (tetrahydrofuran-3-yl)methanamine (0.016 mL, 0.15 mmol) in 1,2-DCE (2 mL) and NMP (0.5 mL) under argon. The resulting solution was stirred at 25° C. for 18 h. The reaction mixture was diluted with DCM (15 mL), water (5 mL) and saturated Na2CO3 (5 mL). The layers were separated and the aqueous layer was extracted with DCM (3×15 mL). The combined organic phase was dried over MgSO4, filtered and evaporated to dryness to give the crude product. The resulting residue was purified by flash silica chromatography, elution gradient 0 to 40% 20% MeOH in DCM (with 1% NH4OH) in DCM. The product fractions were concentrated to dryness under reduced pressure to give 1-(heptadecan-9-yl) 17-(heptan-3-yl) 9-(((tetrahydrofuran-3-yl)methyl)amino)heptadecanedioate (29.8 mg, 71.2%) as a colorless oil. 1 H NMR (500MHz, methanol-d4) δ ppm 0.9-0.9(m,12H)1.3(br s,44H)1.4-1.4(m,4H)1.5(br s,13H)2.0-2.1(m,1H)2.3(td,J=7.1,2.7Hz,4H)2.3-2.4(m,1H)2.5(br t,J=5.7Hz,1H)2.6(br d,J=7.2Hz,2H)3.4-3.5(m,1H)3.7(q,J=7.7Hz,1H)3.8-3.9(m,2H)4.8-4.8(m,1H)4.9-4.9(m,1H);C 46 H 89 NO5 m / z calculated value 735.674 Measured value 736.8 [M+H] + (LCMS).

[0255] Scheme 23 below illustrates the synthetic procedure for preparing Example 48. In step aa below, when the respective amine was utilized as the free base, AcOH was used as an additive in the reductive amination.

[0256] Scheme 23: [ka] Reagents:z)EDC.HCl, DIPEA, DMAP, DCM;aa)NaBH(OAc)3, AcOH, 1,2-DCE:NMP(4:1)

[0257] Example 48. Synthesis of Compound 48: 1-(heptadecan-9-yl) 17-(heptan-2-yl) 9-(((tetrahydrofuran-3-yl)methyl)amino)heptadecanedioate Step z): To a stirred solution of 17-(heptadecan-9-yloxy)-9,17-dioxoheptadecanoic acid (64.9 mg, 0.12 mmol), N-ethyl-N-isopropylpropan-2-amine (0.074 mL, 0.42 mmol), N,N-dimethylpyridin-4-amine (2.151 mg, 0.02 mmol) and heptan-2-ol (0.042 mL, 0.29 mmol) in DCM (3 mL) under argon at 0° C. was added 3-(((ethylimino)methylene)amino)-N,N-dimethylpropan-1-amine hydrochloride (58.5 mg, 0.31 mmol) in one portion. The resulting solution was stirred at 25° C. for 16 h. The reaction mixture was diluted with EtOAc (20 mL), water (5 mL) and saturated NaCl solution (15 mL). The layers were separated and the aqueous layer was extracted with EtOAc (3x20 mL). The combined organic layers were washed with saturated aqueous NaCl (20 mL). The organic layers were dried over MgSO4, filtered and concentrated to dryness under reduced pressure to give the crude product. The resulting residue was purified by flash silica chromatography, elution gradient 0 to 35% EtOAc in hexanes. The product fractions were concentrated to dryness under reduced pressure to give 1-(heptadecan-9-yl) 17-(heptan-2-yl) 9-oxoheptadecanedioate (64.0 mg, 84%) as a colorless oil. 1 H NMR(500MHz,chloroform-d)δ ppm 0.8-0.9(m,9H)1.2-1.2(m,3H)1.2-1.3(m,42H)1.5-1.6(m,14H)2.2-2.3(m,4H)2.4(t,J=7.5Hz,4H)4.9(td,J=13.1,6.3Hz,2H).

[0258] Step aa): [ka] (After 10 min) Sodium triacetoxyhydroborate (58.3 mg, 0.28 mmol) was added in one portion to a stirred solution of 1-(heptadecan-9-yl) 17-(heptan-2-yl) 9-oxoheptadecanedioate (64 mg, 0.10 mmol) and (tetrahydrofuran-3-yl)methanamine (0.027 mL, 0.27 mmol) in 1,2-DCE (2 mL) and NMP (0.5 mL) under argon. The resulting solution was stirred at 25° C. for 18 h. The reaction mixture was diluted with DCM (15 mL), water (5 mL) and saturated Na2CO3 (5 mL). The layers were separated and the aqueous layer was extracted with DCM (3×15 mL). The combined organic phase was dried over MgSO4, filtered and evaporated to dryness to give the crude product. The resulting residue was purified by flash silica chromatography, elution gradient 0 to 40% 20% MeOH in DCM (with 1% NH4OH) in DCM. The product fractions were concentrated to dryness under reduced pressure to give 1-(heptadecan-9-yl) 17-(heptan-2-yl) 9-(((tetrahydrofuran-3-yl)methyl)amino)heptadecanedioate (52.5 mg, 72.5%) as a colorless oil. 1 H NMR(500MHz, methanol-d4)δ ppm 0.9(s,9H)1.2-1.2(m,3H)1.3-1.4(m,46H)1.4-1.5(m,4H)1.5-1.7(m,11H)2.1-2.1(m,1H)2.3-2.3(m,4H)2.4-2.4(m,1H)2.5(br C 46 H 89 NO5 m / z calculated value 735.674 Measured value 736.8 [M+H] + (LCMS).

[0259] Scheme 24 below illustrates the synthetic procedure for preparing Example 49. In steps aj below, when the respective amine was utilized as a free base, AcOH was used as an additive in the reductive amination.

[0260] Scheme 24: [ka] Reagents: ab) DMP, DCM, NaHCO3; ac) BnBr, NaH, THF; ad) Mg, I 2、 THF;ae)DMP, DCM, NaHCO3;af)H2, Pd / C, THF;ag)DMP, DCM, NaHCO3;ah)NaClO2, NaH2PO4, amylene, THF, t-butanol;ai)EDC.HCl, DIPEA, DMAP, DCM;aj)NaBH(OAc)3, AcOH, 1,2-DCE:NMP(4:1)

[0261] Example 49. Synthesis of Compound 49: Bis(3-pentyloctyl) 6-((2-oxaspiro[3.3]heptan-6-yl)amino)hexadecanedioate Step ab): To a stirred suspension of sodium bicarbonate (1670 mg, 19.88 mmol) and 6-(benzyloxy)hexan-1-ol (460 mg, 2.21 mmol) in DCM (15 mL) at 0 °C was added Dess-Martin periodinane (2810 mg, 6.63 mmol) in one portion. The resulting solution was allowed to warm to 25 °C for 5 h. The reaction mixture was diluted with DCM (20 mL) and washed successively with saturated aqueous NaHCO3 (20 mL) and saturated Na2S2O3 (20 mL). The organic layer was dried over MgSO4, filtered and concentrated to dryness under reduced pressure to give the crude product. The resulting residue was purified by flash silica chromatography, elution gradient 0 to 30% EtOAc in hexanes. The product fractions were concentrated to dryness under reduced pressure to give 6-(benzyloxy)hexanal (229 mg, 50.2%) as a colorless liquid. 1 H NMR(500MHz,chloroform-d)1.40-1.49(m,2H),1.66(br d,J=7.6Hz,4H),2.42-2.49(m,2H),3.48(t,J=6.5Hz,2H),4.51(s,2H),7.28-7.32(m,1H),7.32-7.37(m,4H),9.77(t,J=1.7Hz,1H).

[0262] Step ac): To a stirred solution of 10-bromodecan-1-ol (0.872 mL, 4.01 mmol) and (bromomethyl)benzene (0.714 mL, 6.01 mmol) in tetrahydrofuran (9 mL) at 0° C. under argon was added sodium hydride (529 mg, 13.22 mmol) in portions. The resulting suspension was stirred at 25° C. for 20 h. The reaction mixture was quenched with saturated aqueous NaHCO3 (10 mL), diluted with water (10 mL), extracted with DCM (3×20 mL), the organic layer was dried over MgSO4, filtered and concentrated under reduced pressure to give the crude product. The resulting residue was purified by flash silica chromatography, elution gradient 0 to 30% EtOAc in hexanes. The product fractions were concentrated to dryness under reduced pressure to give (((10-bromodecyl)oxy)methyl)benzene (1136 mg, 87%) as a colorless oil. 1 H NMR (500 MHz, chloroform-d) 1.27-1.32 (m, 8H), 1.34-1.45 (m, 4H), 1.58-1.66 (m, 2H), 1.86 (quin, J=7.2 Hz, 2H), 3.39-3.49 (m, 4H), 4.51 (s, 2H), 7.28-7.33 (m, 1H), 7.34-7.36 (m, 4H).

[0263] Step ad): To a stirred suspension of magnesium (134 mg, 5.50 mmol) and (((10-bromodecyl)oxy)methyl)benzene (600 mg, 1.83 mmol) in THF (5 mL) at 25° C. under argon was added iodine (11.63 mg, 0.05 mmol) in one portion. The reaction mixture was heated to 55° C. for 0.5 h. At this point the color of the reaction mixture changes from brown to cloudy white. 6-(benzyloxy)hexanal (189 mg, 0.92 mmol) was dissolved in 2 mL of THF and added dropwise to the solution at 25° C. The reaction mixture was then warmed to 60° C. for 2 h and allowed to reach 25° C. under argon for 15 h. The reaction mixture was quenched and diluted with water (2 mL) followed by the addition of 1 M HCl (10 mL) and DCM (15 mL). The layers were separated and the aqueous layer was extracted with DCM (3x15 mL). The combined organic layers were washed with saturated aqueous NaCl (15 mL). The organic layers were dried over MgSO4, filtered and concentrated to dryness under reduced pressure to give the crude product. TLC indicates the formation of a new, less polar product (Rf = 0.6; 4:1, hexanes:EtOAc). The resulting residue was purified by flash silica chromatography, elution gradient 0 to 55% hexanes in EtOAc. The product fractions were concentrated to dryness under reduced pressure to give 1,16-bis(benzyloxy)hexadecan-6-ol (310 mg, 74.4%) as a colorless oil. 1 H NMR(500MHz,chloroform-d)1.28(br s,9H),1.32-1.51(m,14H),1.59-1.69(m,4H),3.48(td,J=6.5,4.5Hz,4H),3.58(br dd,J=7.1,4.0Hz,1H),4.51(s,4H),7.28-7.31(m,2H),7.33-7.36(m,8H).

[0264] Step ae): To a stirred suspension of sodium bicarbonate (344 mg, 4.09 mmol) and 1,16-bis(benzyloxy)hexadecan-6-ol (310 mg, 0.68 mmol) in DCM (10 mL) at 0 °C was added Dess-Martin periodinane (578 mg, 1.36 mmol) in one portion. The resulting solution was allowed to warm to 25 °C for 24 h. The reaction mixture was diluted with DCM (20 mL) and washed successively with saturated aqueous NaHCO3 (20 mL) and saturated Na2S2O3 (20 mL). The organic layer was dried over MgSO4, filtered, and concentrated to dryness under reduced pressure to give the crude product. The resulting residue was purified by flash silica chromatography, elution gradient 0 to 30% EtOAc in hexanes. The product fractions were concentrated to dryness under reduced pressure to give 1,16-bis(benzyloxy)hexadecan-6-one (270 mg, 87%) as a colorless residue. 1 H NMR(500MHz,chloroform-d)1.20-1.27(m,10H),1.29-1.36(m,4H),1.47-1.61(m,8H),2.27-2.3 5(m,4H),3.41(t,J=6.6Hz,4H),4.43(d,J=4.1Hz,4H),7.17-7.25(m,2H),7.25-7.31(m,8H).

[0265] Step af): 1,16-bis(benzyloxy)hexadecan-6-one (270 mg, 0.60 mmol), palladium 10% on carbon (127 mg, 0.12 mmol) in THF (6 mL) was stirred under hydrogen atmosphere at 25° C. for 16 h. The reaction mixture was filtered through Celite. Evaporation of the solvent gave a precipitate to give 1,16-dihydroxyhexadecan-6-one (168 mg, 104%) as a white solid***. 1 H NMR (500 MHz, chloroform-d) 1.29 (br s, 10H), 1.34-1.41 (m, 4H), 1.54-1.70 (m, 10H), 2.41 (dt, J=15.0, 7.4 Hz, 4H), 3.63-3.68 (m, 4H).

[0266] Steps ag, ah, ai): To a stirred suspension of sodium bicarbonate (466 mg, 5.55 mmol) and 1,16-dihydroxyhexadecan-6-one (168 mg, 0.62 mmol) in DCM (10 mL) at 0 °C was added Dess-Martin periodinane (785 mg, 1.85 mmol) in one portion. The resulting solution was allowed to warm to 25 °C for 24 h. The reaction mixture was diluted with DCM (20 mL) and washed successively with saturated aqueous NaHCO3 (20 mL) and saturated Na2S2O3 (20 mL). The organic layer was dried over MgSO4, filtered, and concentrated to dryness under reduced pressure to give 6-oxohexadecanedial as a colorless dry film, which was used without further purification.

[0267] To a stirred solution of 2-methyl-2-butene (2.83 mL, 26.71 mmol), sodium dihydrogen phosphate (641 mg, 5.34 mmol) and sodium chlorite (5.34 mL, 5.34 mmol) in THF (15 mL) and t-butanol (7.50 mL) at 25° C. was added 6-oxohexadecanedial (239 mg, 0.89 mmol). The resulting solution was stirred at 25° C. for 4 h. The reaction mixture was diluted with DCM and water (30 ml each). The reaction mixture was adjusted to pH=3 with 1 M HCl solution. The organic layer was dried over MgSO4, filtered and concentrated to dryness under reduced pressure to give the desired product 6-oxohexadecanedioic acid as a white solid, which was used without further purification.

[0268] To a stirred solution of 6-oxohexadecanedioic acid (186.3 mg, 0.62 mmol), 3-pentyloctan-1-ol (373 mg, 1.86 mmol), N-ethyl-N-isopropylpropan-2-amine (0.486 mL, 2.79 mmol) and N,N-dimethylpyridin-4-amine (11.37 mg, 0.09 mmol) in DCM (8 mL) at 0° C. under argon was added 3-(((ethylimino)methylene)amino)-N,N-dimethylpropan-1-amine hydrochloride (392 mg, 2.05 mmol) in one portion. The resulting solution was stirred at 25° C. for 18 h. The reaction mixture was diluted with DCM (10 mL) and water (10 mL). The layers were separated and the aqueous layer was extracted with DCM (3×15 mL). The combined organic layers were washed with saturated aqueous NaCl (15 mL). The organic layer was dried over MgSO4, filtered and concentrated to dryness under reduced pressure to give the crude product. The resulting residue was purified by flash silica chromatography, elution gradient 0 to 30% EtOAc in hexanes. The product fractions were concentrated to dryness under reduced pressure to give bis(3-pentyloctyl) 6-oxohexadecandioate (116 mg, 28.2%, 3 steps) as a colorless oil. 1 H NMR (500 MHz, chloroform-d) 0.88-0.93 (m, 16H), 1.24-1.37 (m, 44H), 1.55-1.66 (m, 8H), 2.28-2.34 (m, 4H), 2.37-2.46 (m, 4H), 4.10 (t, J = 7.0 Hz, 4H).

[0269] Steps a-j): [ka] (After 10 min) sodium triacetoxyhydroborate (111 mg, 0.52 mmol) was added in one portion to a stirred solution of bis(3-pentyloctyl) 6-oxohexadecandioate (116.3 mg, 0.17 mmol) and 2-oxaspiro[3.3]heptan-6-aminium chloride (76 mg, 0.51 mmol) in 1,2-DCE (2.4 mL) and NMP (0.5 mL) under argon. The resulting solution was stirred at 25 °C for 40 h. The reaction mixture was diluted with DCM (15 mL), water (5 mL) and saturated Na2CO3 (5 mL). The layers were separated and the aqueous layer was extracted with DCM (3x15 mL). The combined organic phase was dried over MgSO4, filtered and evaporated to dryness to give the crude product. The resulting residue was purified by flash silica chromatography, elution gradient 0 to 40% 20% MeOH in DCM (with 1% NH4OH) in DCM. The product fractions were concentrated to dryness under reduced pressure to give bis(3-pentyloctyl) 6-((2-oxaspiro[3.3]heptan-6-yl)amino)hexadecandioate (18.6 mg, 13.95%) as a colorless oil. 1 H NMR (500MHz, methanol-d4)0.93(t,J=7.0Hz,12H),1.28-1.67(m,60H),2.11(ddd,J=12.6,8.4,4.0Hz,2H),2.34(dt,J=18.8,7 C 48 H 91 NO5 m / z Calculated value 761.690 Measured value 762.9 [M+H] + (LCMS).

[0270] Scheme 25 below illustrates the synthetic procedure for preparing Example 50.

[0271] Scheme 25: [ka] Reagents: a) EDC.HCl, DIPEA, DMAP, DCM; b) NaBH(OAc)3, 1,2-DCE:NMP(4:1)

[0272] Example 50. Synthesis of Compound 50 Step 1: 1-(dodecan-4-yl) 17-(heptadecan-9-yl) 9-oxoheptadecanedioate To a stirred solution of 17-(heptadecan-9-yloxy)-9,17-dioxoheptadecanoic acid (262 mg, 0.47 mmol), N-ethyl-N-isopropylpropan-2-amine (0.289 mL, 1.66 mmol), N,N-dimethylpyridin-4-amine (11.58 mg, 0.09 mmol) and dodecan-4-ol (106 mg, 0.57 mmol) in DCM (10 mL) at 0° C. under argon was added 3-(((ethylimino)methylene)amino)-N,N-dimethylpropan-1-amine hydrochloride (118 mg, 0.62 mmol) in one portion. The resulting solution was stirred at RT for 16 h. The reaction mixture was diluted with DCM (20 mL) and 10% citric acid solution (25 mL). The organic layer was separated and the aqueous layer was extracted with (DCM) (3×25 mL). The combined organic layers were washed with saturated aqueous NaCl (20 mL). The organic layers were dried over MgSO4, filtered, and concentrated to dryness under reduced pressure to give the crude product. The resulting residue was purified by flash silica chromatography, elution gradient 0 to 20% EtOAc in hexanes. The product fractions were concentrated to dryness under reduced pressure to give 1-(dodecan-4-yl) 17-(heptadecan-9-yl) 9-oxoheptadecanedioate (213 mg, 62.3%) as a colorless oil. 1 H NMR (500MHz, chloroform-d, 27℃) δ ppm 0.82-0.98(12H,m),1.18-1.68(66H,m),2.28(4H,t),2.34-2.43(4H,t),4.76-4.97(2H,m).

[0273] Compound 50: 1-(dodecan-4-yl) 17-(heptadecan-9-yl) 9-((2-oxaspiro[3.3]heptan-6-yl)amino)heptadecanedioate [ka] To a stirred solution of 2-oxaspiro[3.3]heptan-6-amine hydrochloride (14.94 mg, 0.10 mmol) and 1-(dodecan-4-yl)17-(heptadecan-9-yl)9-oxoheptadecanedioate (30 mg, 0.04 mmol) in DCE (2 mL) and NMP (0.5 mL) at 0° C. under argon was added sodium triacetoxyborohydride (23.80 mg, 0.11 mmol) in one portion. The resulting solution was stirred at room temperature for 16 h. The reaction mixture was diluted with DCM (50 mL) and saturated sodium carbonate (50 mL). The layers were separated and the aqueous layer was extracted with (DCM) (3×25 mL). The organic layer was dried over MgSO4, filtered and concentrated to dryness under reduced pressure to give the crude product. The resulting residue was purified by flash silica chromatography, elution gradient 0 to 100% in DCM (20% MeOH and 1% NH4OH in DCM). The product fractions were concentrated to dryness under reduced pressure to give 1-(dodecan-4-yl)17-(heptadecan-9-yl)9-((2-oxaspiro[3.3]heptan-6-yl)amino)heptadecanedioate (10.50 mg, 30.8%) as a colorless oil. 1 H NMR (500MHz, chloroform-d, 27℃) δ ppm 0.91(12H,m),1.23-1.68(70H,m),1.96-2.08(2H,m),2.25-2.36(4H,t),2.51-2.60(3 C52H99NO5 m / z Calculated value 817.752 Actual value 818.90[M+H]+(LCMS).

[0274] Scheme 26 below illustrates the synthetic procedures for preparing Examples 51 and 52.

[0275] Scheme 26: [ka] Reagents: a) EDC.HCl, DIPEA, DMAP, DCM; b) NaBH(OAc)3, 1,2-DCE:NMP(4:1)

[0276] Examples 51 and 52. Synthesis of Compounds 51 and 52 Step 1: 1-(heptadecan-9-yl) 17-((2-hexylcyclopropyl)methyl) 9-oxoheptadecanedioate To a stirred solution of 17-(heptadecan-9-yloxy)-9,17-dioxoheptadecanoic acid (123 mg, 0.22 mmol), N-ethyl-N-isopropylpropan-2-amine (0.081 mL, 0.47 mmol), N,N-dimethylpyridin-4-amine (5.44 mg, 0.04 mmol) and (2-hexylcyclopropyl)methanol (41.7 mg, 0.27 mmol) in DCM (5 mL) under argon at 0° C. was added 3-(((ethylimino)methylene)amino)-N,N-dimethylpropan-1-amine hydrochloride (90 mg, 0.47 mmol) in one portion. The resulting solution was stirred at RT for 16 h. The reaction mixture was diluted with DCM (20 mL) and 10% citric acid solution (25 mL). The organic layer was separated and the aqueous layer was extracted with (DCM) (3x25 mL). The combined organic layers were washed with saturated aqueous NaCl (20 mL). The organic layers were dried over MgSO4, filtered and concentrated to dryness under reduced pressure to give the crude product. The resulting residue was purified by flash silica chromatography, elution gradient 0 to 40% EtOAc in hexanes. The product fractions were concentrated to dryness under reduced pressure to give 1-(heptadecan-9-yl) 17-((2-hexylcyclopropyl)methyl) 9-oxoheptadecanedioate (100 mg, 65.0%) as a colorless oil. 1 H NMR (500 MHz, chloroform-d, 27°C) δ ppm -0.02-0.09 (1H,m), 0.70-0.80 (1H,m), 0.84-0.97 (10H,m), 1.09-1.70 (59H,m), 2.24-2.46 (8H,m), 3.88-4.27 (2H,m), 4.88 (1H,m).

[0277] Compound 51: 1-(heptadecan-9-yl) 17-((2-hexylcyclopropyl)methyl) 9-((oxetan-3-ylmethyl)amino)heptadecanedioate [ka] To a stirred solution of oxetan-3-ylmethanamine hydrochloride (10.73 mg, 0.09 mmol) and 1-(heptadecan-9-yl) 17-((2-hexylcyclopropyl)methyl) 9-oxoheptadecanedioate (25 mg, 0.04 mmol) in DCE (2 mL) and NMP (0.5 mL) at 0° C. under argon was added sodium triacetoxyborohydride (20.70 mg, 0.10 mmol) in one portion. The resulting solution was stirred at room temperature for 16 h. The reaction mixture was diluted with DCM (50 mL) and saturated sodium carbonate (50 mL). The layers were separated and the aqueous layer was extracted with (DCM) (3×25 mL). The organic layer was dried over MgSO4, filtered and concentrated to dryness under reduced pressure to give the crude product. The resulting residue was purified by flash silica chromatography, elution gradient 0 to 100% in DCM (20% MeOH and 1% NH4OH in DCM). The product fractions were concentrated to dryness under reduced pressure to give 1-(heptadecan-9-yl) 17-((2-hexylcyclopropyl)methyl) 9-((oxetan-3-ylmethyl)amino)heptadecanedioate (9.20 mg, 33.4%) as a colorless oil. 1 H NMR (500MHz, methanol-d4, 27℃) δ ppm 0.01-0.09(1H,m),0.72-0.81(1H,m),0.88-0.96(10H,m),1.12-1.73(64H,m),2.33(4H,q),2.54-2.63(1H,m),3.00(2H,m) ),3.10-3.23(1H,m),3.61-3.77(1H,m),3.85-3.97(1H,m),4.22-4.30(1H,m),4.42(2H,t),4.89-4.97(1H,m);C48H91NO5 m / z Calculated value 761.690 Actual value 762.80[M+H]+(LCMS).

[0278] Compound 52: 1-(heptadecan-9-yl) 17-((2-hexylcyclopropyl)methyl) 9-(((tetrahydrofuran-3-yl)methyl)amino)heptadecanedioate [ka] To a stirred solution of (tetrahydrofuran-3-yl)methanamine hydrochloride (11.95 mg, 0.09 mmol) and 1-(heptadecan-9-yl)17-((2-hexylcyclopropyl)methyl)9-oxoheptadecanedioate (25 mg, 0.04 mmol) in DCE (2 mL) and NMP (0.5 mL) at 0° C. under argon was added sodium triacetoxyborohydride (20.70 mg, 0.10 mmol) in one portion. The resulting solution was stirred at room temperature for 16 h. The reaction mixture was diluted with DCM (50 mL) and saturated sodium carbonate (50 mL). The layers were separated and the aqueous layer was extracted with (DCM) (3×25 mL). The organic layer was dried over MgSO4, filtered and concentrated to dryness under reduced pressure to give the crude product. The resulting residue was purified by flash silica chromatography, elution gradient 0 to 100% in DCM (20% MeOH and 1% NH4OH in DCM). The product fractions were concentrated to dryness under reduced pressure to give 1-(heptadecan-9-yl) 17-((2-hexylcyclopropyl)methyl) 9-(((tetrahydrofuran-3-yl)methyl)amino) heptadecanedioate (15.60 mg, 55.6%) as a colorless oil. 1H NMR (500 MHz, methanol-d4, 27oC) δ ppm 1H NMR(500MHz,methanol-d4,27oC)0.01-0.09(1H,m),0.71-0.82(1H,m),0.87- 0.98(10H,m),1.13-1.72(64H,m),2.08-2.18(1H,m),2.33(4H,m),2.41-2. 50(1H,m),2.63-2.70(1H,m),2.70-2.75(2H,m),3.47-3.55(1H,m),3.70-3 .81(1H,m),3.89(3H,m),4.22-4.30(1H,m),4.89-4.93(1H,m).;C49H93NO5 m / z calculated value 775.705 Found 776.90[M+H]+(LCMS).

[0279] Scheme 27 below illustrates the synthetic procedure for preparing Example 53.

[0280] Scheme 27: [ka] Reagent:a)TFAA;b)NaBH(OAc)3,1,2-DCE:NMP(4:1)

[0281] Example 53. Synthesis of Compound 53 Step 1: 1-(heptadecan-9-yl) 17-(2-methyldecan-2-yl) 9-oxoheptadecanedioate To a solution of 17-(heptadecan-9-yloxy)-9,17-dioxoheptadecanoic acid (28 mg, 0.05 mmol) in DCM (2 mL) at 0° C. was added TFAA (0.016 mL, 0.11 mmol) dropwise. After 2.5 h, 2-methyldecan-2-ol (31.4 mg, 0.18 mmol) was added slowly. After 1 h, the reaction was warmed to rt and allowed to stir for 2.5 h. The reaction was then quenched with water and extracted with diethyl ether. The organic layer was separated, dried over MgSO4, filtered, and concentrated to dryness under reduced pressure to give the crude product. The residue was purified by silica gel chromatography using EtOAc in hexanes (0-10%) to give 1-(heptadecan-9-yl) 17-(2-methyldecan-2-yl) 9-oxoheptadecanedioate (24.50 mg, 68.4%) as a pale yellow oil. 1 H NMR (500 MHz, chloroform-d, 27° C.) δ ppm 0.85-0.95 (9H, t), 1.22-1.37 (48H, m), 1.43 (6H, s), 1.58 (14H, m), 2.18-2.24 (2H, t), 2.26-2.31 (2H, t), 2.36-2.42 (4H, t), 4.82-4.95 (1H, m).

[0282] Compound 53: 1-(heptadecan-9-yl) 17-(3-methylnonyl) 9-(((tetrahydro-2H-pyran-4-yl)methyl)amino)heptadecanedioate [ka] To a stirred solution of 2-oxaspiro[3.3]heptan-6-amine hydrochloride (12.44 mg, 0.08 mmol) and 1-(heptadecan-9-yl)17-(2-methyldecan-2-yl)9-oxoheptadecanedioate (24.5 mg, 0.03 mmol) in DCE (2 mL) and NMP (0.5 mL) at 0° C. under argon was added sodium triacetoxyborohydride (19.83 mg, 0.09 mmol) in one portion. The resulting solution was stirred at room temperature for 16 h. The reaction mixture was diluted with DCM (50 mL) and saturated sodium carbonate (50 mL). The layers were separated and the aqueous layer was extracted with (DCM) (3×25 mL). The organic layer was dried over MgSO4, filtered and concentrated to dryness under reduced pressure to give the crude product. The resulting residue was purified by flash silica chromatography, elution gradient 0 to 100% in DCM (20% MeOH and 1% NH4OH in DCM). The product fractions were concentrated to dryness under reduced pressure to give 1-(heptadecan-9-yl) 17-(2-methyldecan-2-yl) 9-((2-oxaspiro[3.3]heptan-6-yl)amino)heptadecanedioate (18.20 mg, 65.3%) as a colorless oil. 1 H NMR (500MHz, methanol-d4, 27℃) δ ppm 0.92(9H,t),1.25-1.70(70H,m),1.73-1.82(2H,m),1.96-2.06(2H,m),2.20-2.26(2H,m),2.30-2.36(2H,m),2 .49-2.54(1H,m),2.54-2.60(2H,m),3.18-3.27(1H,m),4.60(2H,s),4.73(2H,s),4.88-4.93(1H,m);C50H97NO5 m / z Calculated value 803.737 Actual value 803.90[M+H]+(LCMS).

[0283] Scheme 28 below illustrates the synthetic procedure for preparing Example 54.

[0284] Scheme 28: [ka] Reagents: a) Mg, I2, THF; b) pyridine sulfur trioxide, TEA; c) TBAF, THF; d) DMP, NaHCO3, DCM and then NaClO2, t-BuOH, 2-methyl-2-butene, NaH2PO4; e) EDC.HCl, DIPEA, DMAP, DCM; f) Pd / C, H2; g) EDC.HCl, DIPEA, DMAP, DCM; h) NaBH(OAc)3, 1,2-DCE:NMP (4:1).

[0285] Example 54. Synthesis of Compound 54 Step 1: 16-(benzyloxy)-1-((triisopropylsilyl)oxy)hexadecan-8-ol To a suspension of Mg turnings in THF (15 mL) containing a small iodine crystal was added a few drops of the appropriate brominated compound (1 equiv.) in THF (0.5 mL / mmol substrate). The mixture was heated until the reaction started, then (((7-bromoheptyl)oxy)methyl)benzene (0.589 g, 2.07 mmol) was added dropwise to maintain gentle reflux unassisted. After complete addition of the starting material, the mixture was heated under reflux for 1 h. The solution of Grignard reagent was cooled and titrated before use. 9-((triisopropylsilyl)oxy)nonanal (0.500 g, 1.59 mmol) was added in one portion to the stirred mixture under argon. The resulting mixture was stirred at 70 °C for 16 h. The reaction mixture was quenched with water (50 mL) and extracted with DCM (3x25 mL) and the organic layer was dried over MgSO4, filtered and concentrated to dryness under reduced pressure to give a pale yellow oil. The resulting residue was purified by flash silica chromatography, elution gradient 0 to 20% EtOAc in hexanes. The product fractions were concentrated to dryness under reduced pressure to give 1-(benzyloxy)-16-((triisopropylsilyl)oxy)hexadecan-8-ol (0.392 g, 47.3%) as a pale yellow oil. 1 H NMR (500MHz, chloroform-d, 27℃) δ ppm 1.04-1.14(21H,m),1.33(22H,m),1.51-1.57(2H,m),1.60-1.67(2H,m),3.44-3. 52(2H,t),3.58-3.63(1H,m),3.65-3.71(2H,t),4.53(2H,s),7.29-7.40(5H,m).

[0286] Step 2: 1-(benzyloxy)-16-((triisopropylsilyl)oxy)hexadecan-8-one In an oven-dried flask, 1-(benzyloxy)-16-((triisopropylsilyl)oxy)hexadecan-8-ol (0.392 g, 0.75 mmol) was dissolved in DCM (10 mL). To the reaction mixture was then added DMSO (2.000 mL) followed by TEA (1.049 mL, 7.53 mmol). The mixture was cooled to 0° C. Pyridine-sulfur trioxide (1 / 1) (0.958 g, 6.02 mmol) was added to the mixture and the reaction was allowed to warm to room temperature. The reaction mixture was stirred at room temperature for 1 h. The reaction mixture was diluted with DCM and the reaction mixture was quenched with saturated aqueous NH4Cl (100 mL). The layers were separated, the aqueous layer was extracted with EtOAc (3×50 mL), and the combined organic layers were washed with brine (50 mL), dried over MgSO4, filtered, and concentrated to dryness under reduced pressure to give a pale yellow oil. The resulting residue was purified by flash silica chromatography, elution gradient 0 to 20% EtOAc in hexanes. The product fractions were concentrated to dryness under reduced pressure to give 1-(benzyloxy)-16-((triisopropylsilyl)oxy)hexadecan-8-one (0.200 g, 51.2%) as a colorless oil. 1 H NMR (500 MHz, chloroform-d, 27° C.) δ ppm 1.02-1.13 (21H, m), 1.24-1.45 (14H, m), 1.50-1.70 (8H, m), 2.34-2.45 (4H, t), 3.43-3.53 (2H, t), 3.62-3.76 (2H, t), 4.47-4.55 (2H, s), 7.36 (5H, m).

[0287] Step 3: 1-(benzyloxy)-16-hydroxyhexadecan-8-one To a stirred solution of 1-(benzyloxy)-16-((triisopropylsilyl)oxy)hexadecan-8-one (0.200 g, 0.39 mmol) in THF (5 mL) at 0° C. under argon was added TBAF (1.542 mL, 1.54 mmol) dropwise. The resulting mixture was stirred at RT for 16 h. The reaction mixture was quenched with saturated aqueous NH4Cl (50 mL), extracted with EtOAc (3×50 mL), and the organic layer was dried over MgSO4, filtered, and concentrated to dryness under reduced pressure to give an orange oil. The resulting residue was purified by flash silica chromatography, elution gradient 0 to 40% EtOAc in hexanes. The product fractions were concentrated to dryness under reduced pressure to give 1-(benzyloxy)-16-hydroxyhexadecan-8-one (0.124 g, 89%) as a colorless oil. 1 H NMR (500 MHz, chloroform-d, 27° C.) δ ppm 1.23-1.70 (22H, m), 2.40 (4H, t), 3.48 (2H, t), 3.66 (2H, t), 4.52 (2H, s), 7.30-7.42 (5H, m).

[0288] Step 4: 16-(benzyloxy)-9-oxohexadecanoic acid i) To a stirred suspension of sodium bicarbonate (259 mg, 3.08 mmol) and 1-(benzyloxy)-16-hydroxyhexadecan-8-one (124 mg, 0.34 mmol) in DCM (5 mL) at 0 °C was added Dess-Martin periodinane (435 mg, 1.03 mmol) in one portion. The resulting solution was allowed to come to room temperature over 24 h. The reaction mixture was diluted with DCM (20 mL) and washed successively with saturated aqueous NaHCO3 (20 mL) and saturated Na2S2O3 (20 mL). The organic layer was dried over MgSO4, filtered, and concentrated to dryness under reduced pressure to give the crude aldehyde precursor as a colorless dry film, which was used without further purification.

[0289] ii) The crude product was added to a stirred solution of 2-methyl-2-butene (1.087 mL, 10.26 mmol), sodium dihydrogen phosphate (246 mg, 2.05 mmol) and sodium chlorite (186 mg, 2.05 mmol) in THF (10 mL) and tert-butanol (5.00 mL) at 25° C. The resulting solution was stirred at RT for 4 h. The reaction mixture was diluted with DCM and water (30 ml each). The reaction mixture was adjusted to pH=3 with 1 M HCl solution. The organic layer was dried over MgSO4, filtered and concentrated to dryness under reduced pressure to give the crude product. The resulting residue was purified by flash silica chromatography, elution gradient 0 to 100% EtOAc in hexanes. The product fractions were concentrated to dryness under reduced pressure to give 16-(benzyloxy)-9-oxohexadecanoic acid (126 mg, 98%) as a white solid. 1 H NMR (500 MHz, chloroform-d, 27° C.) δ ppm 1.24-1.45 (12H, m), 1.57 (8H, m), 2.30-2.44 (6H, m), 3.48 (2H, t), 4.52 (2H, s), 7.26-7.41 (5H, m).

[0290] Step 5: Heptadecane-9-yl 16-(benzyloxy)-9-oxohexadecanoate To a stirred mixture of 16-(benzyloxy)-9-oxohexadecanoic acid (125.9 mg, 0.33 mmol), heptadecan-9-ol (111 mg, 0.43 mmol), DIPEA (0.123 mL, 0.70 mmol) and DMAP (8.17 mg, 0.07 mmol) in DCM (5 mL) at 0° C. under argon was added EDC (109 mg, 0.57 mmol) in one portion. The resulting mixture was stirred at room temperature for 16 h. The reaction mixture was diluted with 10% citric acid (25 mL) and DCM (25 mL). The layers were separated and the aqueous layer was extracted with (DCM) (4×25 mL). The combined organic layers were dried over MgSO4, filtered and concentrated to dryness under reduced pressure to give the crude product. The resulting residue was purified by flash silica chromatography, elution gradient 0 to 40% EtOAc in hexanes. The product fractions were concentrated to dryness under reduced pressure to give heptadecan-9-yl 16-(benzyloxy)-9-oxohexadecanoate (126 mg, 61.3%) as a colorless oil. 1 H NMR (500 MHz, chloroform-d, 27° C.) δ ppm 0.90 (6H, t), 1.28 (48H, m), 2.26-2.32 (2H, t), 2.34-2.43 (4H, t), 3.39-3.54 (2H, t), 4.52 (2H, s), 4.89 (1H, m), 7.28-7.39 (5H, m).

[0291] Step 6: Heptadecane-9-yl 16-hydroxy-9-oxohexadecanoate Heptadecane-9-yl 16-(benzyloxy)-9-oxohexadecanoate (126 mg, 0.20 mmol) and Pd / C (65.4 mg, 0.06 mmol) were stirred in MeOH (5 mL) under hydrogen atmosphere at ambient pressure and RT for 16 h. The reaction mixture was filtered through Celite. The resulting residue was purified by flash silica chromatography, elution gradient 0 to 40% EtOAc in hexanes. The product fractions were concentrated to dryness under reduced pressure to give heptadecane-9-yl 16-hydroxy-9-oxohexadecanoate (95 mg, 88%) as a colorless oil. 1 H NMR (500 MHz, chloroform-d, 27° C.) δ ppm 0.90 (6H, t), 1.22-1.69 (48H, m), 2.25-2.33 (2H, t), 2.40 (4H, t), 3.66 (2H, t), 4.82-4.93 (1H, m).

[0292] Step 7: Heptadecane-9-yl 16-(decanoyloxy)-9-oxohexadecanoate To a stirred mixture of heptadecan-9-yl 16-hydroxy-9-oxohexadecanoate (50 mg, 0.10 mmol), decanoic acid (0.028 mL, 0.14 mmol), DIPEA (0.068 mL, 0.39 mmol) and DMAP (2.328 mg, 0.02 mmol) in DCM (5 mL) at 0° C. under argon was added EDC (38.4 mg, 0.20 mmol) in one portion. The resulting mixture was stirred at room temperature for 16 h. The reaction mixture was diluted with saturated sodium bicarbonate (25 mL) and DCM (25 mL). The layers were separated and the aqueous layer was extracted with (DCM) (4×25 mL). The combined organic layers were dried over MgSO4, filtered and concentrated to dryness under reduced pressure to give the crude product. The resulting residue was purified by flash silica chromatography, elution gradient 0 to 40% EtOAc in hexanes. The product fractions were concentrated to dryness under reduced pressure to give heptadecan-9-yl 16-(decanoyloxy)-9-oxohexadecanoate (45.0 mg, 69.6%) as a colorless oil. 1 H NMR (500 MHz, chloroform-d, 27° C.) δ ppm 0.89 (9H, m), 1.20-1.42 (48H, m), 1.47-1.69 (14H, m), 2.23-2.33 (4H, t), 2.35-2.44 (4H, t), 3.98-4.13 (2H, t), 4.81-4.94 (1H, m).

[0293] Compound 54: Heptadecane-9-yl 9-((2-oxaspiro[3.3]heptan-6-yl)amino)-16-(decanoyloxy)hexadecanoate [ka] To a stirred solution of heptadecan-9-yl 16-(decanoyloxy)-9-oxohexadecanoate (45 mg, 0.07 mmol) and 2-oxaspiro[3.3]heptan-6-amine hydrochloride (23.79 mg, 0.16 mmol) in DCE (2 mL) and NMP (0.5 mL) at 0° C. under argon was added sodium triacetoxyborohydride (37.9 mg, 0.18 mmol) in one portion. The resulting solution was stirred at room temperature for 16 h. The reaction mixture was diluted with DCM (50 mL) and saturated sodium carbonate (50 mL). The layers were separated and the aqueous layer was extracted with (DCM) (3×25 mL). The organic layer was dried over MgSO4, filtered and concentrated to dryness under reduced pressure to give the crude product. The resulting residue was purified by flash silica chromatography, elution gradient 0 to 100% in DCM (20% MeOH and 1% NH4OH in DCM). The product fractions were concentrated to dryness under reduced pressure to give heptadecan-9-yl 9-((2-oxaspiro[3.3]heptan-6-yl)amino)-16-(decanoyloxy)hexadecanoate (37.5 mg, 72.9%) as a colorless oil. 1 H NMR (500MHz, methanol-d4, 27℃) δ ppm 0.93(9H,t),1.32(56H,m),1.51-1.70(10H,m),1.96-2.03(2H,m),2.33(4H,t),2.48-2.54(1H,m),2.54-2.61(2 C49H93NO5 m / z Calculated value 776.705 Actual value 778.80[M+H]+(LCMS).

[0294] Scheme 29 below illustrates the synthetic procedure for preparing Example 55.

[0295] Scheme 29: [ka] Reagents: a) Mg, I2, THF; b) pyridine sulfur trioxide, TEA; c) TBAF, THF; d) DMP, NaHCO3, DCM and then NaClO2, t-BuOH, 2-methyl-2-butene, NaH2PO4; e) EDC.HCl, DIPEA, DMAP, DCM; f) Pd / C, H2; g) DMP, NaHCO3, DCM and then NaClO2, t-BuOH, 2-methyl-2-butene, NaH2PO4; h) EDC.HCl, DIPEA, DMAP, DCM; i) NaBH(OAc)3, 1,2-DCE:NMP (4:1).

[0296] Example 55. Synthesis of Compound 55 Step 1: 1-(benzyloxy)-19-((triisopropylsilyl)oxy)nonadecan-9-ol To a suspension of magnesium (0.284 g, 11.67 mmol) turnings in DMF (20 mL) containing small iodine crystals was added a few drops of the appropriate brominated compound in THF (10 mL). The mixture was heated until the reaction started, then the remaining (((8-bromooctyl)oxy)methyl)benzene (2.096 g, 7.00 mmol) was added dropwise, maintaining a gentle reflux without assistance. After complete addition of the starting material, the mixture was heated under reflux for 1 h. The solution of Grignard reagent was cooled and titrated before use. 11-((triisopropylsilyl)oxy)undecanal (2 g, 5.84 mmol) was added in one portion to the stirred mixture under argon. The resulting mixture was stirred at 70 °C for 16 h. The reaction mixture was quenched with water (50 mL) and extracted with DCM (3x25 mL) and the organic layer was dried over MgSO4, filtered and concentrated to dryness under reduced pressure to give a pale yellow oil. The resulting residue was purified by flash silica chromatography, elution gradient 0 to 40% EtOAc in hexanes. The product fractions were concentrated to dryness under reduced pressure to give 1-(benzyloxy)-19-((triisopropylsilyl)oxy)nonadecan-9-ol (1.837 g, 74.4%) as a colorless oil. 1 H NMR (500MHz, chloroform-d, 27℃) δ ppm 1.03-1.14(21H,m),1.26-1.47(28H,m),1.53-1.58(2H,m),1.60-1.67(2H,m),3. 44-3.54(2H,t),3.54-3.63(1H,m),3.65-3.73(2H,t),4.53(2H,s),7.36(5H,m).

[0297] Step 2: 1-(benzyloxy)-19-((triisopropylsilyl)oxy)nonadecan-9-one In an oven dried flask, 1-(benzyloxy)-19-((triisopropylsilyl)oxy)nonadecan-9-ol (2.477 g, 4.40 mmol) was dissolved in DCM (80 mL). To the reaction mixture was then added DMSO (15 mL) followed by TEA (6.13 mL, 44.00 mmol). The mixture was cooled to RT. Pyridine sulfur trioxide was added to the mixture and the reaction was allowed to warm to room temperature. The reaction mixture was stirred at room temperature for 1 h. The reaction mixture was diluted with DCM and the reaction mixture was quenched with saturated aqueous NH4Cl (100 mL). The layers were separated, the aqueous layer was extracted with EtOAc (3x50 mL) and the combined organic layers were washed with brine (50 mL), dried over MgSO4, filtered and concentrated to dryness under reduced pressure to give a pale yellow oil. The resulting residue was purified by flash silica chromatography, elution gradient 0 to 20% EtOAc in hexanes. The product fractions were concentrated to dryness under reduced pressure to give 1-(benzyloxy)-19-((triisopropylsilyl)oxy)nonadecan-9-one (1.837 g, 74.4%) as a colorless oil. 1 H NMR (500 MHz, chloroform-d, 27° C.) δ ppm 1.03-1.18 (21H, m), 1.29 (28H, m), 2.33-2.44 (4H, t), 3.42-3.55 (2H, t), 3.62-3.73 (2H, t), 4.52 (2H, s), 7.36 (5H, m).

[0298] Step 3: 1-(benzyloxy)-19-hydroxynonadecan-9-one Tetrabutylammonium fluoride (13.10 mL, 13.10 mmol) was added dropwise to a stirred solution of 1-(benzyloxy)-19-((triisopropylsilyl)oxy)nonadecan-9-one (1.837 g, 3.27 mmol) in THF (10 mL) at 0° C. under argon. The resulting mixture was stirred at RT for 16 h. The reaction mixture was quenched with saturated aqueous NH4Cl (50 mL), extracted with EtOAc (3×50 mL), and the organic layer was dried over MgSO4, filtered, and concentrated to dryness under reduced pressure to give an orange oil. The resulting residue was purified by flash silica chromatography, elution gradient 0 to 40% EtOAc in hexanes. The product fractions were concentrated to dryness under reduced pressure to give 1-(benzyloxy)-19-hydroxynonadecan-9-one (1.266 g, 96%) as a colorless oil. 1 H NMR (500 MHz, chloroform-d, 27° C.) δ ppm 1.23-1.70 (30H, m), 2.40 (4H, t), 3.48 (2H, t), 3.66 (2H, t), 4.52 (2H, s), 7.30-7.42 (5H, m).

[0299] Step 4: 19-(benzyloxy)-11-oxonadecanoic acid i) To a stirred suspension of sodium bicarbonate (2.365 g, 28.16 mmol) and 1-(benzyloxy)-19-hydroxynonadecan-9-one (1.266 g, 3.13 mmol) in DCM (20 mL) at 0° C. was added 3-oxo-115-benzo[d][1,2]iodaoxole-1,1,1(3H)-triyl triacetate (3.98 g, 9.39 mmol) in one portion. The resulting solution was allowed to come to room temperature over 24 h. The reaction mixture was diluted with DCM (20 mL) and washed successively with saturated aqueous NaHCO3 (20 mL) and saturated Na2S2O3 (20 mL). The organic layer was dried over MgSO4, filtered, and concentrated to dryness under reduced pressure to give the crude aldehyde precursor as a colorless dry film, which was used without further purification.

[0300] ii) The crude product was added to a stirred solution of 2-methylbut-2-ene (9.94 mL, 93.86 mmol), sodium dihydrogen phosphate (2.252 g, 18.77 mmol) and sodium chlorite (1.698 g, 18.77 mmol) in THF (10 mL) and tert-butanol (5.00 mL) at 25° C. The resulting solution was stirred at RT for 4 h. The reaction mixture was diluted with DCM and water (30 ml each). The reaction mixture was adjusted to pH=3 with 1M HCl solution. The organic layer was dried over MgSO4, filtered and concentrated to dryness under reduced pressure to give the crude product. The resulting residue was purified by flash silica chromatography, elution gradient 0 to 100% EtOAc in hexane. The product fractions were concentrated to dryness under reduced pressure to give the product 19-(benzyloxy)-11-oxononadecanoic acid (1.275 g, 97%) as a white solid. 1 H NMR (500 MHz, chloroform-d, 27° C.) δ ppm 1.30 (26H, m), 2.28-2.50 (6H, m), 3.37-3.60 (2H, t), 4.53 (2H, s), 7.25-7.38 (5H, m).

[0301] Step 5: Octan-2-yl 19-(benzyloxy)-11-oxonadecanoate To a stirred mixture of 19-(benzyloxy)-11-oxononadecanoic acid (400 mg, 0.96 mmol), octan-2-ol (0.195 mL, 1.24 mmol), DIPEA (0.350 mL, 2.01 mmol) and DMAP (23.35 mg, 0.19 mmol) in DCM (5 mL) at 0° C. under argon was added EDC (311 mg, 1.62 mmol) in one portion. The resulting mixture was stirred at room temperature for 16 h. The reaction mixture was diluted with saturated sodium bicarbonate (25 mL) and DCM (25 mL). The layers were separated and the aqueous layer was extracted with (DCM) (4×25 mL). The combined organic layers were dried over MgSO4, filtered and concentrated to dryness under reduced pressure to give the crude product. The resulting residue was purified by flash silica chromatography, elution gradient 0 to 40% EtOAc in hexanes. The product fractions were concentrated to dryness under reduced pressure to give octan-2-yl 19-(benzyloxy)-11-oxononadecanoate (410 mg, 81%) as a colorless oil. 1 H NMR (500MHz, chloroform-d, 27℃) δ ppm 0.83-0.97(3H,m),1.15-1.67(39H,m),2.18-2.31(2H,m),2.39(4H,t) ,3.39-3.54(2H,m),4.52(2H,s),4.80-5.00(1H,m),7.28-7.40(5H,m).

[0302] Step 6: Octan-2-yl 19-hydroxy-11-oxonadecanoate Octan-2-yl 19-(benzyloxy)-11-oxononadecanoate (410 mg, 0.77 mmol) and Pd / C (247 mg, 0.23 mmol) in MeOH (10 mL) were stirred under hydrogen atmosphere and at RT for 16 h. The resulting residue was purified by flash silica chromatography, elution gradient 0 to 60% EtOAc in hexanes. The product fractions were concentrated to dryness under reduced pressure to give octan-2-yl 19-hydroxy-11-oxononadecanoate (200 mg, 58.8%) as a pale yellow oil. 1 H NMR (500 MHz, chloroform-d, 27° C.) δ ppm 0.92 (3H, t), 1.17-1.68 (39H, m), 2.22-2.32 (2H, m), 2.40 (4H, t), 3.55-3.67 (2H, m), 4.90 (1H, m).

[0303] Step 7: 19-(octan-2-yloxy)-9,19-dioxononadecanoic acid i) DMP (577 mg, 1.36 mmol) was added in one portion to a stirred suspension of sodium bicarbonate (343 mg, 4.08 mmol) and octan-2-yl 19-hydroxy-11-oxononadecanoate (200 mg, 0.45 mmol) in DCM (5 mL) at 0° C. The resulting solution was allowed to come to room temperature over 24 h. The reaction mixture was diluted with DCM (20 mL) and washed successively with saturated aqueous NaHCO3 (20 mL) and saturated Na2S2O3 (20 mL). The organic layer was dried over MgSO4, filtered, and concentrated to dryness under reduced pressure to give the crude aldehyde precursor as a colorless dry film, which was used without further purification.

[0304] ii) The crude product was added to a stirred solution of 2-methylbut-2-ene (1.442 mL, 13.61 mmol), sodium dihydrogen phosphate (327 mg, 2.72 mmol) and sodium chlorite (246 mg, 2.72 mmol) in THF (10 mL) and tert-butanol (5.00 mL) at 25° C. The resulting solution was stirred at RT for 4 h. The reaction mixture was diluted with DCM and water (30 ml each). The reaction mixture was adjusted to pH=3 with 1 M HCl solution. The organic layer was dried over MgSO4, filtered and concentrated to dryness under reduced pressure to give the crude product. The resulting residue was purified by flash silica chromatography, elution gradient 0 to 100% EtOAc in hexanes. The product fractions were concentrated to dryness under reduced pressure to give the desired product 19-(octan-2-yloxy)-9,19-dioxononadecanoic acid as a white solid. 1 H NMR (500 MHz, chloroform-d, 27° C.) δ ppm 0.92 (3H, t), 1.24-1.72 (38H, m), 2.29 (2H, t), 2.35-2.47 (6H, m), 4.87 (1H, m).

[0305] Step 8: 1-(heptadecan-9-yl) 19-(octan-2-yl) 9-oxonadecanedioate To a stirred mixture of 19-(octan-2-yloxy)-9,19-dioxononadecanoic acid (256 mg, 0.56 mmol), heptadecan-9-ol (217 mg, 0.84 mmol), DIPEA (0.403 mL, 2.31 mmol) and DMAP (13.76 mg, 0.11 mmol) in DCM (5 mL) at 0° C. under argon was added EDC (227 mg, 1.18 mmol) in one portion. The resulting mixture was stirred at room temperature for 16 h. The reaction mixture was diluted with saturated sodium bicarbonate (25 mL) and DCM (25 mL). The layers were separated and the aqueous layer was extracted with (DCM) (4×25 mL). The combined organic layers were dried over MgSO4, filtered and concentrated to dryness under reduced pressure to give the crude product. The resulting residue was purified by flash silica chromatography, elution gradient 0 to 40% EtOAc in hexanes. The product fractions were concentrated to dryness under reduced pressure to give 1-(heptadecan-9-yl) 19-(octan-2-yl) 9-oxonadecandioate (141 mg, 36.1%) as a colorless oil. 1 H NMR (500 MHz, chloroform-d, 27° C.) δ ppm 0.86-0.95 (9H, m), 1.18-1.69 (65H, m), 2.29 (4H, m), 2.35-2.44 (4H, m), 4.80-4.98 (2H, m).

[0306] Compound 55: 1-(heptadecan-9-yl) 19-(octan-2-yl) 9-((2-oxaspiro[3.3]heptan-6-yl)amino)nonadecanedioate [ka] To a stirred solution of 1-(heptadecan-9-yl) 19-(octan-2-yl) 9-oxononadecanedioate (0.141 g, 0.20 mmol) and 2-oxaspiro[3.3]heptan-6-amine hydrochloride (0.073 g, 0.49 mmol) in DCE (2 mL) and NMP (0.5 mL) at 0° C. under argon was added sodium triacetoxyborohydride (0.116 g, 0.55 mmol) in one portion. The resulting solution was stirred at room temperature for 16 h. The reaction mixture was diluted with DCM (50 mL) and saturated sodium carbonate (50 mL). The layers were separated and the aqueous layer was extracted with (DCM) (3×25 mL). The organic layer was dried over MgSO4, filtered and concentrated to dryness under reduced pressure to give the crude product. The resulting residue was purified by flash silica chromatography, elution gradient 0 to 100% in DCM (20% MeOH and 1% NH4OH in DCM). The product fractions were concentrated to dryness under reduced pressure to give 1-(heptadecan-9-yl) 19-(octan-2-yl) 9-((2-oxaspiro[3.3]heptan-6-yl)amino)nonadecandioate (0.099 g, 61.7%) as a colorless oil. 1 H NMR (500MHz, methanol-d4, 27℃) δ ppm 0.92(9H,t),1.18-1.71(71H,m),1.91-2.04(2H,m),2.27-2.37(4H,m),2.43-2.50(1H ,m),2.52-2.59(2H,m),3.11-3.23(1H,m),4.56-4.64(2H,s),4.73(2H,s);C49H93NO5 m / z Calculated value 789.721 Actual value 790.80[M+H]+(LCMS).

[0307] Scheme 30 below illustrates the synthetic procedure for preparing Example 56.

[0308] Scheme 30: [ka] Reagents: a) I2, THF; b) pyridine sulfur trioxide, TEA; c) TBAF, THF; d) DMP, NaHCO3, DCM and then NaClO2, t-BuOH, 2-methyl-2-butene, NaH2PO4; e) EDC.HCl, DIPEA, DMAP, DCM; f) Pd / C, H2; g) DMP, NaHCO3, DCM and then NaClO2, t-BuOH, 2-methyl-2-butene, NaH2PO4; h) EDC.HCl, DIPEA, DMAP, DCM; i) NaBH(OAc)3, 1,2-DCE:NMP (4:1).

[0309] Example 56. Synthesis of Compound 56 Step 1: 1-(benzyloxy)-19-((triisopropylsilyl)oxy)nonadecan-9-ol (6-(benzyloxy)hexyl)magnesium bromide (5.79 mL, 2.89 mmol) was diluted in THF (10 mL) containing a small crystal of iodine. The solution of Grignard reagent was cooled to 0° C. 9-((triisopropylsilyl)oxy)nonanal (0.7 g, 2.23 mmol) was added in one portion to the stirred mixture at 0° C. under argon. The resulting mixture was stirred at 70° C. for 16 h. The reaction mixture was quenched with water (50 mL), extracted with DCM (3×25 mL), and the organic layer was dried over MgSO4, filtered, and concentrated to dryness under reduced pressure to give a pale yellow oil. The resulting residue was purified by flash silica chromatography, elution gradient 0 to 40% EtOAc in hexanes. The product fractions were concentrated to dryness under reduced pressure to give 1-(benzyloxy)-15-((triisopropylsilyl)oxy)pentadecan-7-ol (0.550 g, 49%) as a colorless oil. 1 H NMR (500MHz, chloroform-d, 27℃) δ ppm 1.04-1.13(21H,m),1.25-1.49(20H,m),1.51-1.60(2H,m),1.60-1.69(2H,m),3. 43-3.53(2H,t),3.55-3.63(1H,m),3.66-3.73(2H,t),4.53(2H,s),7.36(5H,m).

[0310] Step 2: 1-(benzyloxy)-19-((triisopropylsilyl)oxy)nonadecan-9-one In an oven dried flask, 1-(benzyloxy)-15-((triisopropylsilyl)oxy)pentadecan-7-ol (0.860 g, 1.70 mmol) was dissolved in DCM (40 mL). To the reaction mixture was then added DMSO (7.50 mL) followed by TEA (2.365 mL, 16.97 mmol). The mixture was cooled to 0° C. Pyridine sulfur trioxide (2.160 g, 13.57 mmol) was added to the mixture and the reaction was allowed to warm to room temperature. The reaction mixture was stirred at room temperature for 1 h. The reaction mixture was diluted with DCM and the reaction mixture was quenched with saturated aqueous NH4Cl (100 mL). The layers were separated, the aqueous layer was extracted with EtOAc (3×50 mL) and the combined organic layers were washed with brine (50 mL), dried over MgSO4, filtered and concentrated to dryness under reduced pressure to give a pale yellow oil. The resulting residue was purified by flash silica chromatography, elution gradient 0 to 20% EtOAc in hexanes. The product fractions were concentrated to dryness under reduced pressure to give 1-(benzyloxy)-15-((triisopropylsilyl)oxy)pentadecan-7-one (0.684 g, 80%) as a colorless oil. 1 H NMR (500MHz, chloroform-d, 27℃) δ ppm 0.98-1.15(21H,m),1.25-1.71(20H,m),2.32-2.50(4H,m),3.42-3.52(2H,t),3.62-3.74(2H,t),4.52(2H,s),7.36(5H,m).

[0311] Step 3: 1-(benzyloxy)-15-hydroxypentadecan-7-one Tetrabutylammonium fluoride (5.42 mL, 5.42 mmol) was added dropwise to a stirred solution of 1-(benzyloxy)-15-((triisopropylsilyl)oxy)pentadecan-7-one (0.684 g, 1.35 mmol) in THF (10 mL) at 0° C. under argon. The resulting mixture was stirred at RT for 16 h. The reaction mixture was quenched with saturated aqueous NH4Cl (50 mL), extracted with EtOAc (3×50 mL), and the organic layer was dried over MgSO4, filtered, and concentrated to dryness under reduced pressure to give an orange oil. The resulting residue was purified by flash silica chromatography, elution gradient 0 to 40% EtOAc in hexanes. The product fractions were concentrated to dryness under reduced pressure to give 1-(benzyloxy)-15-hydroxypentadecan-7-one (0.385 g, 82%) as a colorless oil. 1 H NMR (500 MHz, chloroform-d, 27° C.) δ ppm 1.23-1.70 (20H, m), 2.40 (4H, t), 3.48 (2H, t), 3.66 (2H, t), 4.52 (2H, s), 7.30-7.42 (5H, m).

[0312] Step 4: 15-(benzyloxy)-9-oxopentadecanoic acid i) To a stirred suspension of sodium bicarbonate (0.900 g, 10.72 mmol) and 1-(benzyloxy)-15-hydroxypentadecan-7-one (0.415 g, 1.19 mmol) in DCM (10 mL) at 0° C. was added 3-oxo-115-benzo[d][1,2]iodaoxole-1,1,1(3H)-triyl triacetate (1.515 g, 3.57 mmol) in one portion. The resulting solution was allowed to come to room temperature over 24 h. The reaction mixture was diluted with DCM (20 mL) and washed successively with saturated aqueous NaHCO3 (20 mL) and saturated Na2S2O3 (20 mL). The organic layer was dried over MgSO4, filtered, and concentrated to dryness under reduced pressure to give the crude aldehyde precursor as a colorless dry film, which was used without further purification.

[0313] ii) The crude product was added to a stirred solution of 2-methylbut-2-ene (3.78 mL, 35.72 mmol), sodium dihydrogen phosphate (0.857 g, 7.14 mmol) and sodium chlorite (0.646 g, 7.14 mmol) in THF (10.00 mL) and tBuOH (5 mL) at 25° C. The resulting solution was stirred at RT for 4 h. The reaction mixture was diluted with DCM and water (30 ml each). The reaction mixture was adjusted to pH=3 with 1 M HCl solution. The organic layer was dried over MgSO4, filtered and concentrated to dryness under reduced pressure to give the crude product. The resulting residue was purified by flash silica chromatography, elution gradient 0 to 100% EtOAc in hexanes. The product fractions were concentrated to dryness under reduced pressure to give the desired product 15-(benzyloxy)-9-oxopentadecanoic acid (0.433 g, 100%) as a white solid. 1 H NMR (500 MHz, chloroform-d, 27° C.) δ ppm 1.29 (18H, m), 2.39 (6H, m), 3.48 (2H t), 4.52 (2H, s), 7.30-7.42 (5H, m).

[0314] Step 5: Heptadecan-9-yl 15-(benzyloxy)-9-oxopentadecanoate To a stirred mixture of 15-(benzyloxy)-9-oxopentadecanoic acid (433 mg, 1.19 mmol), heptadecan-9-ol (398 mg, 1.55 mmol), DIPEA (0.438 mL, 2.51 mmol) and DMAP (29.2 mg, 0.24 mmol) in DCM (5 mL) at 0° C. under argon was added EDC (389 mg, 2.03 mmol) in one portion. The resulting mixture was stirred at room temperature for 16 h. The reaction mixture was diluted with saturated sodium bicarbonate (25 mL) and DCM (25 mL). The layers were separated and the aqueous layer was extracted with (DCM) (4×25 mL). The combined organic layers were dried over MgSO4, filtered and concentrated to dryness under reduced pressure to give the crude product. The resulting residue was purified by flash silica chromatography, elution gradient 0 to 40% EtOAc in hexanes. The product fractions were concentrated to dryness under reduced pressure to give heptadecan-9-yl 15-(benzyloxy)-9-oxopentadecanoate (276 mg, 38.4%) as a colorless oil. 1 H NMR (500 MHz, chloroform-d, 27° C.) δ ppm 0.83-0.95 (6H, t), 1.21-1.70 (46H, m), 2.29 (2H, t), 2.36-2.45 (4H, m), 3.42-3.53 (2H, t), 4.52 (2H, s), 4.89 (1H, m), 7.35 (5H, m).

[0315] Step 6: Heptadecan-9-yl 15-hydroxy-9-oxopentadecanoate Heptadecan-9-yl 15-(benzyloxy)-9-oxopentadecanoate (276 mg, 0.46 mmol) and Pd / C (147 mg, 0.14 mmol) in MeOH (10 mL) were stirred under hydrogen atmosphere at RT for 16 h. The resulting residue was purified by flash silica chromatography, elution gradient 0 to 60% EtOAc in hexanes. The product fractions were concentrated to dryness under reduced pressure to give heptadecan-9-yl 15-hydroxy-9-oxopentadecanoate as a pale yellow oil. 1 H NMR (500 MHz, chloroform-d, 27° C.) δ ppm 0.92 (6H, t), 1.23-1.71 (46H, m), 2.24-2.32 (2H, t), 2.37-2.45 (4H, m), 3.54-3.70 (2H, m), 4.87 (1H, m).

[0316] Step 7: 15-(benzyloxy)-9-oxopentadecanoic acid i) To a stirred suspension of sodium bicarbonate (281 mg, 3.35 mmol) and heptadecan-9-yl 15-hydroxy-9-oxopentadecanoate (190 mg, 0.37 mmol) in DCM (10 mL) at 0° C., 3-oxo-115-benzo[d][1,2]iodaoxole-1,1,1(3H)-triyl triacetate (473 mg, 1.12 mmol) was added in one portion. The resulting solution was allowed to come to room temperature over 24 h. The reaction mixture was diluted with DCM (20 mL) and washed successively with saturated aqueous NaHCO3 (20 mL) and saturated Na2S2O3 (20 mL). The organic layer was dried over MgSO4, filtered, and concentrated to dryness under reduced pressure to give the crude aldehyde precursor as a colorless dry film, which was used without further purification.

[0317] ii) The crude product was added to a stirred solution of 2-methylbut-2-ene (1.182 mL, 11.16 mmol), sodium dihydrogen phosphate (268 mg, 2.23 mmol) and sodium bicarbonate (281 mg, 3.35 mmol) in THF (10.00 mL) and tBuOH (5 mL) at 25° C. The resulting solution was stirred at RT for 4 h. The reaction mixture was diluted with DCM and water (30 ml each). The reaction mixture was adjusted to pH=3 with 1 M HCl solution. The organic layer was dried over MgSO4, filtered and concentrated to dryness under reduced pressure to give the crude product. The resulting residue was purified by flash silica chromatography, elution gradient 0 to 100% EtOAc in hexanes. The product fractions were concentrated to dryness under reduced pressure to give 15-(benzyloxy)-9-oxopentadecanoic acid (0.433 g, 100%) as a white solid. 1 H NMR (500 MHz, chloroform-d, 27° C.) δ ppm 0.92 (6H, t), 1.24-1.72 (44H, m), 2.29 (2H, t), 2.35-2.47 (6H, m), 4.87 (1H, m).

[0318] Step 8: 1-(dodecan-2-yl) 15-(heptadecan-9-yl) 7-oxopentadecane diacetate To a stirred mixture of 15-(heptadecan-9-yloxy)-7,15-dioxopentadecanoic acid (200 mg, 0.38 mmol), dodecan-2-ol (0.128 mL, 0.57 mmol), DIPEA (0.273 mL, 1.56 mmol) and DMAP (9.31 mg, 0.08 mmol) in DCM (5 mL) at 0° C. under argon was added EDC (153 mg, 0.80 mmol) in one portion. The resulting mixture was stirred at room temperature for 16 h. The reaction mixture was diluted with saturated sodium bicarbonate (25 mL) and DCM (25 mL). The layers were separated and the aqueous layer was extracted with (DCM) (4×25 mL). The combined organic layers were dried over MgSO4, filtered and concentrated to dryness under reduced pressure to give the crude product. The resulting residue was purified by flash silica chromatography, elution gradient 0 to 40% EtOAc in hexanes. The product fractions were concentrated to dryness under reduced pressure to give 1-(dodecan-2-yl) 15-(heptadecan-9-yl) 7-oxopentadecandioate as a colorless oil. 1 H NMR (500 MHz, chloroform-d, 27° C.) δ ppm 0.90 (9H, t), 1.17-1.72 (66H, m), 2.29 (4H, t), 2.40 (4H, m), 4.90 (1H, m).

[0319] Compound 56: 1-(dodecan-2-yl) 15-(heptadecan-9-yl) 7-((2-oxaspiro[3.3]heptan-6-yl)amino)pentadecanedioate [ka] To a stirred solution of 1-(dodecan-2-yl) 15-(heptadecan-9-yl) 7-oxopentadecandioate (125 mg, 0.18 mmol) and 2-oxaspiro[3.3]heptan-6-amine hydrochloride (64.8 mg, 0.43 mmol) in DCE (2 mL) and NMP (0.5 mL) at 0° C. under argon was added sodium triacetoxyborohydride (103 mg, 0.49 mmol) in one portion. The resulting solution was stirred at room temperature for 16 h. The reaction mixture was diluted with DCM (50 mL) and saturated sodium carbonate (50 mL). The layers were separated and the aqueous layer was extracted with (DCM) (3×25 mL). The organic layer was dried over MgSO4, filtered and concentrated to dryness under reduced pressure to give the crude product. The resulting residue was purified by flash silica chromatography, elution gradient 0 to 100% in DCM (20% MeOH and 1% NH4OH in DCM). The product fractions were concentrated to dryness under reduced pressure to give 1-(dodecan-2-yl)15-(heptadecan-9-yl)7-((2-oxaspiro[3.3]heptan-6-yl)amino)pentadecanedioate (85 mg, 59.9%) as a colorless oil. 1 H NMR (500MHz, methanol-d4, 27℃) δ ppm 0.92(9H,t),1.31(71H,m),1.91-2.05(2H,m),2.26-2.37(4H,m),2.43-2.50(1H,m),2 .52-2.60(2H,m),3.11-3.23(1H,m),4.53-4.63(2H,s),4.69-4.78(2H,s);C49H93NO5 m / z Calculated value 789.721 Actual value 790.80[M+H]+(LCMS).

[0320] Scheme 31 below shows the synthetic procedure for preparing Example 57.

[0321] Scheme 31: [ka] Reagents: a) NaBH(OAc)3, AcOH, 1,2-DCE:NMP (4:1)

[0322] Example 57. Synthesis of Compound 57 Compound 57: Bis(3-pentyloctyl) 9-((6-oxaspiro[3.4]octan-2-yl)amino)heptadecanedioate [ka] Bis(3-pentyloctyl) 9-oxoheptadecanedioate was prepared according to the protocol described for Example 1. Sodium triacetoxyhydroborate (76 mg, 0.36 mmol) was then added in one portion to a stirred solution of bis(3-pentyloctyl) 9-oxoheptadecanedioate (90 mg, 0.13 mmol) and 6-oxaspiro[3.4]octan-2-amine (40.5 mg, 0.32 mmol) in DCE (2 mL) and NMP (0.5 mL) at 0° C. under argon. The resulting solution was stirred at room temperature for 16 h. The reaction mixture was diluted with DCM (50 mL) and saturated sodium carbonate (50 mL). The layers were separated and the aqueous layer was extracted with (DCM) (3×25 mL). The organic layer was dried over MgSO4, filtered, and concentrated to dryness under reduced pressure to give the crude product. The resulting residue was purified by flash silica chromatography, elution gradient 0 to 100% in DCM (20% MeOH and 1% NH4OH in DCM). The product fractions were concentrated to dryness under reduced pressure to give bis(3-pentyloctyl) 9-((6-oxaspiro[3.4]octan-2-yl)amino)heptadecanedioate (87 mg, 83%) as a colourless oil. 1 H NMR (500 MHz, methanol-d4, 27 °C) δ ppm 0.93 (12H,t), 1.25-1.69 (62H,m), 1.83-2.04 (4H,m), 2.23-2.36 (6H,m), 2.47-2.57 (1H,m), 3.33-3.44 (1H,m), 3.58-3.73 (2H,m), 3.73-3.84 (2H,m), 4.12 (4H,t).; C50H95NO5 m / z calculated 789.721 found 790.9 [M+H]+ (LCMS).

[0323] Comparative Example 1. Synthesis of [(10Z,13Z)-1-[(9Z,12Z)-octadeca-9,12-dienyl]nonadeca-10,13-dienyl]4-(dimethylamino)butanoate (DLin-MC3-DMA or MC3) [ka] DLin-MC3-DMA (MC3) was prepared according to the method described in International Publication No. 2010144740 (Example 5, p. 140). 1 H NMR (400 MHz, CDCl3) δ 5.27 - 5.45 (m, 8H), 4.81 - 4.93 (m, 1H), 2.78 (t, 4H), 2.32 (q, 4H), 2.24 (s, 6H), 2.05 (q, 8H), 1.81 (q, 2H), 1.44 - 1.59 (m, 4H), 1.21 - 1.45 (m, 36H), 0.90 (t, 6H). Number of expected H: 79; assigned H: 79. LCMS m / z 642.5 [M+H] + .

[0324] Comparative Example 2. Synthesis of heptadecan-9-yl 8-((2-hydroxyethyl)(8-(nonyloxy)-8-oxooctyl)amino)octanoate (MOD5) [ka] MOD5 was prepared according to the procedure for lipid 5 in Sabnis et al. (Mol Ther. 2018, 26(6), 1509-1519).

[0325] Comparative Example 3. Synthesis of heptadecan-9-yl 8-((2-hydroxyethyl)(6-oxo-6-(undecyloxy)hexyl)amino)octanoate (MOD8) [ka] MOD8 was prepared according to the procedure for lipid 8 in Sabnis et al. (Mol Ther. 2018, 26(6), 1509-1519).

[0326] Example 58. Preparation of lipid nanoparticle (LNP) formulations A solution of eGFP mRNA (purchased from TriLink Biotechnologies) in citrate buffer was prepared by mixing mRNA dissolved in MilliQ water, 100 mM citrate buffer (pH 3) and MilliQ water to obtain a solution of 50 mM citrate. A lipid solution (99.5%) in ethanol was prepared with four different lipid components: ionizable lipid (see Table 1); cholesterol (Sigma-Aldrich); DSPC (distearoylphosphatidylcholine, Avanti Polar Lipid Inc); and polymer-bound lipid (see Table 1). The lipid ratio in all experiments was ionizable lipid / cholesterol / DSPC / polymer-bound lipid (50 / 38.5 / 10 / 1.5 mol%). The total lipid concentration in all experiments was 12.5 mM.

[0327] The mRNA and lipid solutions were mixed in a NanoAssemblr (Precision Nanosystems, Vancouver, BC, Canada) microfluidic mixing system at a mixing ratio of 3:1 aqueous solution:EtOH and a constant flow rate of 12 mL / min. The mRNA in the citrate buffer solution was prepared so that, upon mixing, the ratio between the nitrogen atoms on the ionizable lipids and the phosphorus atoms on the mRNA chain (N / P ratio) was either 3:1 or 6:1 (see Table 1).

[0328] The first 0.2-0.35 mL and the last 0.05-0.1 mL of the prepared LNP suspension were discarded, while the remainder of the volume was collected as sample fractions. The size of the mRNA-lipid nanoparticles was determined by dynamic light scattering measurements using a Zetasizer Nano ZS from Malvern Instruments Ltd, which directly gives the z-average particle size. A particle refractive index of 1.45 was used to calculate the number-based particle size distribution and mean.

[0329] The final mRNA concentration and encapsulation efficiency percentage (%EE) were measured by Quant-it Ribogreen Assay Kit (ThermoFischer Scientific Inc.) using Triton-X100 to destroy LNPs. The mRNA encapsulation efficiency was determined according to the following formula:

number

[0330] Table 1 summarizes the characteristics of LNP formulations containing Compound 1 or MC3.

[0331] [Table 1]

[0332] DMPE-PEG2000 is dimyristoylphosphatidylethanolamine-poly(ethylene glycol) 2000 (obtained from NOF Corporation). DMG-PEG2000 is 1,2-dimyristoyl-rac-glycero-3-methoxypolyethylene glycol-2000.

[0333] Example 59. In vitro expression of eGFP in 16HBE cells In vitro expression of EGFP protein from LNP formulations 1 and 7 described in Example 58 was tested in the human bronchial-epithelial cell line 16HBE (Sigma-Aldrich SCC150). 16HBE cells were maintained in DMEM, low glucose with GlutaMAX™ + pyruvate (Gibco 21885-025) supplemented with MEM non-essential amino acids (Gibco 11140035) and 10% heat-inactivated fetal calf serum (HI-FCS). Cells were cultured at 37°C in a humidified atmosphere containing 5% CO2. The day before the experiment, cells were detached from culture flasks using TrypLE™ (Gibco 12604013) and plated in cell culture-treated 96-well plates (Greiner Bio-One Cells were seeded at a density of 20.000 cells / well in 1000 x g of DMEM containing 1% HI-FCS (#655090). On the day of the experiment, 1 h prior to incubation with LNP, the medium was removed and replaced with 95 μl DMEM containing 1% HI-FCS. After 1 h of acclimation, 5 μl of LNP in PBS was added and mixed by the Bravo robot, resulting in a final concentration of 50-125 ng mRNA / well. Cells were then incubated for 24 h at 37°C in a humidified atmosphere containing 5% CO2. Absolute quantification of EGFP protein was performed using ELISA (GFP SimpleStep ELISA kit, Abcam #ab171581). After 24 h of incubation, cells were lysed by adding 100 μl of lysis buffer to each well (lysis buffer included in the kit or Abcam product numbers ab193970 and ab193971) and then the cells were lysed by freeze-thaw cycles in lysis buffer. Lysates were measured in the appropriate dilutions according to the kit manufacturer's instructions. Results were 125 ng Reported as the number of EGFP molecules expressed per administered mRNA after 24 hours, calculated from the mRNA / well values. Results are plotted as mean + SEM from triplicate values. Results are summarized in Table 2.

[0334] The expression of eGFP in 16HBE cytosol proves the function of the LNP composition used in eGFP mRNA in promoting gene transfection and transcription. Quantification of eGFP allows the comparison of different LNP compositions with a lipid composition based on MC3 used as a comparator in terms of induction of protein expression in 16HBE to obtain lipid to MC3 ratios. In this application, the LNP compositions differ in terms of their ionizable lipid (IL) components. Thus, the expression results show the potential of the ionizable lipids described herein for application in gene therapy. (See FIG. 1).

[0335] [Table 2]

[0336] Example 60. In vivo intratracheal administration of LNP formulations to rats All experiments were performed in accordance with Swedish Animal Welfare and approved by the Ethical Committee for Laboratory Animals in Gothenburg, Sweden. Male Wistar rats with an average body weight of 250 g were purchased from Charles River Laboratories (Germany). For intratracheal (it) treatment with either PBS or LNP formulations (formulations 1 and 7 in Example 58), rats were anesthetized with an isoflurane mixture (air / oxygen and 4% isoflurane) and instilled using a modified metal cannula with a bolus valve at the top, in a supine position at a 30-40° angle. After it administration, rats were placed in a cage in a supine position with their heads up until consciousness was regained. The instillation volume was 1 ml / kg rat. Twenty-four hours after treatment, rats were sacrificed by ip injection of Allfatal vet (100 mg / ml) and sectioning of the vena cava.

[0337] Determination of cytokine levels in rat BALF Bronchoalveolar lavage (BAL) was performed by manually perfusing the entire lung. After exposing the trachea, a polyethylene tube (PE120) was inserted and ligated with 1-0 silk thread. The tube was connected to a syringe, pre-filled with 4 mL PBS at room temperature, and PBS was slowly infused into the lungs. BAL (BALF) fluid was again collected by slowly aspirating into the syringe, then slowly reinjected into the lungs, and finally removed and transferred to a test tube.

[0338] Tubes containing BALF samples were kept on ice until centrifugation (Hettich ROTANTA 46R, 1200 rpm, 10 min, 4°C). After centrifugation, the supernatant was removed and the cell pellet was resuspended in 0.5 mL PBS, kept on ice, and immediately processed for cell counting. Total and differential numbers of cells were counted using an automated Hematology Analyzer SYSMEX XT-1800i Vet (Sysmex, Kobe Japan). Cell suspensions were vortexed before Sysmex analysis.

[0339] Determination of eGFP protein in lung tissue homogenates The expression of eGFP in rat lungs confirmed the functionality of the applied LNP formulations in inducing the transcription of cargo genes in vivo. The number of eGFP molecules makes it possible to evaluate the efficiency of the LNP formulations containing compound 1 compared to a comparative formulation based on MC3 (see FIG. 1). The inflammatory effect of each of the LNP formulations was evaluated by measuring chemokine release (e.g. neutrophil release) in the BAL. The determination of the increase in chemokine release compared to the vehicle effect was used to compare the LNP formulations with regard to the induction of a deleterious inflammatory response in the treated tissues (see FIG. 2).

[0340] Example 61. In vivo intracardiac administration of LNP formulations to rats Wistar rats were anesthetized with isoflurane and connected to a rat ventilator using a nose cone. Rats were ventilated with ∼1100 ml / min of air and ∼100 ml / min of oxygen (∼60) strokes / min, (∼4 μl tidal volume). Core body temperature was maintained at 37.5 ± 1°C by a heated operating table and a heat lamp controlled by rectal temperature.

[0341] The stomach and thoracic area were shaved and surgically scrubbed. A left thoracotomy was performed at the 5th intercostal space approximately 2-3 mm left of the sternum using scissors. Marcaine 5 ml / kg was given sc on the side of the thoracotomy for local analgesia. A thoracotomy tool was used to hold the incision open. A ligature with 6-0 suture (Prolene) was placed to open the pericardium and allow the heart to be lifted and to mark the injection site. The formulation was injected into the myocardium. The suture was tied with a loose knot, set aside, and the chest was closed by suturing, and the rat was kept warm and on a ventilator until consciousness was regained. Temgesic 10 ml / kg was given sc for long term analgesia before the rat was returned to its cage.

[0342] Each formulation (formulations 2 and 8 from Example 58) was injected three times into rat myocardium using a syringe (Myjector U-100 Insulin, 0.33mm*12mm) and a compound volume of 20μl per injection. The mRNA concentration of the formulation was 0.05mg / mL, so a total dose of 3μg mRNA was given to each animal. N=3 animals / group. 24 hours after dosing, rats were anesthetized with isoflurane and when in the surgical plane of anesthesia, cardiac puncture was performed to collect blood samples and drain the heart from blood. The heart (right ventricle, divided into five 0.5-1g sections) and liver (0.5-1g right lobe) were obtained for protein quantification. All pieces of tissue were weighed and then placed in Precellys tubes, flash frozen in liquid nitrogen, and stored in a -80°C freezer until analysis. Blood was collected in EDTA tubes, placed on ice, and plasma was prepared by centrifugation (3.000 RCF for 10 min at 4° C.) within 30 min of sampling. Plasma was separated into one 50 μl aliquot (haptoglobin) and one 50 μl aliquot (cytokines) and stored in a −80° C. freezer until analysis.

[0343] EGFP quantification was performed in tissue samples by EGFP ELISA (see Figures 3 and 4) and blood samples were used for analysis of haptoglobin and the cytokines; IL-6, MCP-1, IP-10, KC (see Figures 5, 6, 7 and 8).

[0344] Example 62. In vivo intravenous and intramuscular administration of NP formulations to mice Female BALB / c mice were purchased from (SPF (Beijing) Laboratory Animal Technology Co. Ltd.) and housed in cages in groups of four on corncob bedding with standard diet upon arrival and provided with free access to tap water that was purified and autoclaved before feeding to the animals. The environment was maintained at a target temperature of 22 ± °C and relative humidity of 40-80% with a 12-h light / dark cycle. Animals were acclimated to the housing conditions for at least 7 days prior to any experimental procedures and were approximately 6-8 weeks of age at the start of dosing.

[0345] Animals were assigned to individual groups with equal mean body weights for each treatment group, N=4 per treatment group.

[0346] Formulations 1, 3, 4, 5 and 6 from Example 58 were each administered intravenously: each mouse was removed from its cage, restrained and then administered the formulation as a slow IV bolus into the lateral tail vein at a dose volume of 0.3 mg / kg.

[0347] Formulations 1, 5 and 9 from Example 58 were each administered intramuscularly: each mouse was removed from its cage, restrained, and then administered in the caudal thigh area by slowly injecting the formulation into the muscle with a dosing volume of 50 μL. All animals were checked for general condition after dosing to ensure that they did not show any signs of morbidity after treatment.

[0348] Blood samples were collected 6 hours after orbital plexus dosing and 24 hours after dosing at the time of sacrifice by cardiac puncture. Whole blood was collected into EDTA tubes and centrifuged at 4000 rpm for 10 minutes. Plasma was then analyzed for cytokines, CRP and haptoglobin.

[0349] All animals were sacrificed 24 hours after dosing and flowing blood sampling was performed, and livers from animals dosed by IV route and liver and injection site muscle from animals dosed IM were harvested for eGFP analysis by ELISA.

[0350] Figure 9 shows the resulting eGFP expression in the liver 24 hours after intravenous administration of the LNP formulation. Figure 10 shows the resulting eGFP expression in the muscle 24 hours after intramuscular administration of the LNP formulation. Figure 11 shows the resulting eGFP expression in the liver 24 hours after intramuscular administration of the LNP formulation.

[0351] Further LNP formulations were prepared according to the procedure described in Example 46. These formulations were tested using the same protocol as described herein. The results of eGFP expression in the liver 24 hours after intravenous administration of this formulation are summarized in Tables 3, 4, 5 and 6 below.

[0352] [Table 3]

[0353] [Table 4]

[0354] [Table 5]

[0355] [Table 6] < / d> < / d> < / d>

Claims

1. Compounds of formula (I) 【Chemistry 1】 or a pharmaceutically acceptable salt thereof, wherein A is, 【Chemistry 2】 and a and b are each independently 6, 7, or 8; c, d, f, and g are each independently 1 or 2; e is 0, 1 or 2; R １ and R ２ are each independently, 【Transformation 3】 and h is 0, 1, 2 or 3; R ３ and R ４ are each independently -(CH ２ ) ｉ CH ３ and i is 3, 4, 5, 6 or 7).

2. 2. The compound of claim 1, or a pharmaceutically acceptable salt thereof, wherein e is 0 or 1.

3. A is, 【Chemistry 4】 2. The compound of claim 1, selected from:

4. 2. The compound of claim 1, or a pharmaceutically acceptable salt thereof, wherein a is 7.

5. 2. The compound of claim 1, or a pharmaceutically acceptable salt thereof, wherein b is 7.

6. 2. The compound of claim 1, or a pharmaceutically acceptable salt thereof, wherein h is 2.

7. R ３ and R ４ are respectively -(CH ２ ) ４ CH ３ 2. The compound of claim 1, wherein:

8. Formula (II): 【Transformation 5】 2. The compound of claim 1, which is: or a pharmaceutically acceptable salt thereof.

9. Compounds of formula (III): 【Transformation 6】 or a pharmaceutically acceptable salt thereof, wherein A is a 4- to 6-membered monocyclic oxacyclyl or a 6- to 10-membered bicyclic oxacyclyl; L is a covalent bond or C １ -C ３ alkylene; a and b are each independently 5, 6, 7, or 8; R １ and R ２ are each independently C ７ -C １１ Straight chain alkyl or C ９ -C １９ branched alkyl; １ and R ２ At least one of them is C ７ -C １１ (not a straight chain alkyl).

10. Compounds of formula (IIIa): 【Transformation 7】 or a pharmaceutically acceptable salt thereof, wherein: A is a 4- to 6-membered monocyclic oxacyclyl or a 6- to 10-membered bicyclic oxacyclyl; L is a covalent bond or C １ -C ３ alkylene; X １ and X ２ are each independently, 【Transformation 8】 and * indicates R １ Show the connection point for a and b are each independently 4, 5, 6, 7, 8, or 9; provided that when one of a and b is 4 or 5, the other is 6, 7, 8, or 9; R １ and R ２ are each independently C ７ -C １１ Straight chain alkyl, C ７ -C １９ Branched alkyl or C ７ -C １９ alkylene-cyclopropylene-alkyl; １ and R ２ At least one of them is C ７ -C １１ Not a straight chain alkyl or at least one is C ７ -C １９ (Not alkylene-cyclopropylene-alkyl).

11. X １ and X ２ But both 【Chemistry 9】 11. The compound of claim 10, wherein:

12. X １ but, 【Chemistry 10】 and X ２ but, 【Chemistry 11】 11. The compound of claim 10, wherein:

13. A is, 【Chemistry 12】 and c is 0, 1 or 2; d is 1, 2, or 3; provided that the sum of c and d is 2 to 4; f is 0, 1 or 2; g is 1, 2, or 3; and the sum of f and g is 2 to 4.

10. The compound of claim 9 or a pharmaceutically acceptable salt thereof.

14. L is a covalent bond, -CH ２ - or -CH ２ CH ２ 10. The compound of claim 9, wherein: - or a pharmaceutically acceptable salt thereof.

15. R １ and R ２ Both are C ９ -C １９ 10. The compound of claim 9, or a pharmaceutically acceptable salt thereof, wherein the alkyl groups are branched and contain the same number of carbon atoms.

16. R １ But C ７ -C １１ Straight chain alkyl or C ９ -C １９ is a branched alkyl; R ２ is C ９ -C １９ branched alkyl; １ and R ２ 10. The compound of claim 9, or a pharmaceutically acceptable salt thereof, wherein:

17. R １ But C ７ -C １１ is a linear alkyl; R ２ is C ７ -C １９ 11. The compound of claim 10, or a pharmaceutically acceptable salt thereof, which is branched alkyl.

18. R １ But C ７ -C １９ Branched alkyl or C ７ -C １９ alkylene-cyclopropylene-alkyl; R ２ is C ７ -C １９ branched alkyl; １ and R ２ 11. The compound of claim 10, or a pharmaceutically acceptable salt thereof, wherein:

19. R １ But C ７ -C １５ Branched alkyl or C ７ -C １５ alkylene-cyclopropylene-alkyl; R ２ is C １３ -C １９ 20. The compound of claim 18, or a pharmaceutically acceptable salt thereof, which is branched alkyl.

20. 11. The compound of claim 10, wherein a and b are the same and both are 6, 7, or 8, or a pharmaceutically acceptable salt thereof.

21. 11. The compound according to claim 10, or a pharmaceutically acceptable salt thereof, wherein a and b are not the same and each independently represents 4 to 9, with the proviso that (a) when one of a and b is 4 or 5, the other is 6, 7, 8, or 9; and (b) the sum of a and b is 12 to 16.

22. R １ But -(CH ２ ) ｍ -CH ３ or 【Chemistry 13】 and R ２ but, 【Chemistry 14】 and m is 7, 8 or 9; n and h are each independently 0, 1, 2, or 3; R ３ａ and R ４ａ are each independently -(CH ２ ) ｐ CH ３ and R ３ｂ and R ４ｂ are each independently -(CH ２ ) ｑ CH ３ and p and q are each independently 0, 1, 2, 3, 4, 5, 6, 7, 8, or 9; ３ａ and R ４ａ together with the carbon atom to which they are attached contain at least 9 carbon atoms, and R ３ｂ and R ４ｂ together with the carbon atom to which they are attached contain at least 9 carbon atoms; 10. The compound of claim 9 or a pharmaceutically acceptable salt thereof.

23. R １ But -(CH ２ ) ｍ -CH ３ , 【Chemistry 15】 and R ２ but, 【Chemistry 16】 and m is 6, 7, 8 or 9; v is 1, 2 or 3; t is 3, 4, 5, 6, 7, or 8; n and h are each independently 0, 1, 2, or 3; R ３ａ and R ４ａ are each independently -(CH ２ ) ｐ CH ３ and R ３ｂ and R ４ｂ are each independently -(CH ２ ) ｑ CH ３ and R ５ａ is hydrogen or methyl; p and q are each independently 0, 1, 2, 3, 4, 5, 6, 7, 8, or 9; ３ａ , R ４ａ and R ５ａ together with the carbon atom to which they are attached contain at least 7 carbon atoms, and R ３ｂ and R ４ｂ together with the carbon atom to which they are attached contain at least 9 carbon atoms; 11. The compound of claim 10, or a pharmaceutically acceptable salt thereof.

24. R ３ａ But -(CH ２ ) ｐ CH ３ where p is 0, 1, 2 or 3; R ４ａ But -(CH ２ ) ｑ CH ３ 24. The compound of claim 23, wherein p is 4, 5, 6, 7, 8, or 9, or a pharmaceutically acceptable salt thereof.

25. R ３ｂ and R ４ｂ Both are -(CH ２ ) ｑ CH ３ and q is 5, 6, 7 or 8; or a pharmaceutically acceptable salt thereof.

26. R １ but, 【Chemistry 17】 10. The compound of claim 9, selected from: or a pharmaceutically acceptable salt thereof.

27. R １ but, [Chemistry 18] 11. The compound of claim 10, selected from:

28. R ２ but, 【Chemistry 19】 10. The compound of claim 9, selected from: or a pharmaceutically acceptable salt thereof.

29. R ２ but, 【Chemistry 20】 11. The compound of claim 10, selected from:

30. Formula (IV): 【Chemistry 21】 The compound of claim 9, represented by or a pharmaceutically acceptable salt thereof, wherein: A is, 【Chemistry 22】 and a and b are each independently 5, 6, 7, or 8; R １ is C ７ -C １１ Straight chain alkyl or C ９ -C １９ is a branched alkyl; R ２ is C ９ -C １９ is a branched alkyl).

31. Formula (IVa): 【Chemistry 23】 The compound of claim 10, represented by or a pharmaceutically acceptable salt thereof, wherein: A is, 【Chemistry 24】 and a and b are each independently 5, 6, 7, or 8; R １ is C ７ -C １１ Straight chain alkyl, C ７ -C １５ Branched alkyl or C ７ -C １５ alkylene-cyclopropylene-alkyl; R ２ is C １５ -C １９ is a branched alkyl).

32. R １ and R ２ Both are C １０ -C １７ 31. The compound of claim 30, or a pharmaceutically acceptable salt thereof, wherein the alkyl groups are branched and contain the same number of carbon atoms.

33. R １ But C ７ -C １１ Straight chain alkyl or C ９ -C １３ is a branched alkyl; R ２ is C １３ -C １９ branched alkyl; １ and R ２ At least one of them is C １３ 31. The compound of claim 30, or a pharmaceutically acceptable salt thereof, which is not branched alkyl.

34. 31. The compound of claim 30, wherein a and b are both 5, 6, 7, or 8, or a pharmaceutically acceptable salt thereof.

35. Formula (V): 【Chemistry 25】 The compound of claim 9, represented by or a pharmaceutically acceptable salt thereof, wherein: A is, 【Chemistry 26】 and a and b are each independently 5, 6, 7, or 8; R １ is C ７ -C １１ Straight chain alkyl or C ９ -C １９ is a branched alkyl; R ２ is C ９ -C １９ is a branched alkyl).

36. Formula (Va): 【Chemistry 27】 The compound of claim 10, represented by or a pharmaceutically acceptable salt thereof, wherein A is, 【Chemistry 28】 and a and b are each independently 5, 6, 7, or 8; R １ is C ７ -C １５ Branched alkyl or C ７ -C １５ alkylene-cyclopropylene-alkyl; R ２ is C １５ -C １９ is a branched alkyl).

37. R １ and R ２ Both are C １０ -C １７ 36. The compound of claim 35, or a pharmaceutically acceptable salt thereof, wherein the alkyl groups are branched and contain the same number of carbon atoms.

38. R １ But C ７ -C １１ Straight chain alkyl or C ９ -C １３ is a branched alkyl; R ２ But C １３ -C １９ branched alkyl; １ and R ２ At least one of them is C １３ 36. The compound of claim 35, or a pharmaceutically acceptable salt thereof, which is not branched alkyl.

39. 36. The compound of claim 35, wherein a and b are both 5, 6, 7, or 8, or a pharmaceutically acceptable salt thereof.

40. Formula (VI): 【Chemistry 29】 The compound of claim 9, represented by or a pharmaceutically acceptable salt thereof, wherein A is, 【Transformation 30】 is selected from L is a covalent bond, -CH ２ - or -CH ２ CH ２ - and; a and b are each independently 5, 6, 7, or 8; R １ is C ７ -C １１ Straight chain alkyl or C ９ -C １９ is a branched alkyl; R ２ is C ９ -C １９ is a branched alkyl).

41. Formula (VIa): 【Chemistry 31】 The compound of claim 10, represented by or a pharmaceutically acceptable salt thereof, wherein A is, 【Chemistry 32】 Selected from: a and b are each independently 5, 6, 7, or 8; R １ is C ７ -C １５ Branched alkyl or C ７ -C １５ alkylene-cyclopropylene-alkyl; R ２ is C １５ -C １９ is a branched alkyl).

42. R １ and R ２ But both are C １０ -C １７ 41. The compound of claim 40, or a pharmaceutically acceptable salt thereof, wherein the alkyl groups are branched and contain the same number of carbon atoms.

43. R １ But C ７ -C １１ Straight chain alkyl or C ９ -C １３ is a branched alkyl; R ２ But C １３ -C １９ branched alkyl; １ and R ２ At least one of them is C １３ 41. The compound of claim 40, or a pharmaceutically acceptable salt thereof, which is not branched alkyl.

44. 41. The compound of claim 40, wherein a and b are both 5, 6, 7, or 8, or a pharmaceutically acceptable salt thereof.

45. Formula (VII): 【Transformation 33】 The compound of claim 9, represented by or a pharmaceutically acceptable salt thereof, wherein A is, 【Transformation 34】 and a and b are each independently 5, 6, 7, or 8; R １ is C ７ -C １１ Straight chain alkyl or C ９ -C １９ is a branched alkyl; R ２ is C ９ -C １９ is a branched alkyl).

46. Formula (VIIa): 【Chemistry 35】 The compound of claim 10, represented by or a pharmaceutically acceptable salt thereof, wherein: A is, 【Transformation 36】 and a and b are each independently 4, 5, 6, 7, or 8; provided that when one of a and b is 4 or 5, the other is 6, 7, 8, or 9; X １ and X ２ are each independently, 【Chemistry 37】 and * indicates R １ Show the connection point for R １ is C ７ -C １１ Straight chain alkyl or C ７ -C １５ is a branched alkyl; R ２ is C １５ -C １９ is a branched alkyl).

47. 47. The compound of claim 46, or a pharmaceutically acceptable salt thereof, wherein a and b are both 6, 7, or 8; or alternatively, a and b are each independently 4 to 9; with the proviso that (a) when one of a and b is 4 or 5, the other is 6, 7, 8, or 9; or (b) the sum of a and b is 12 to 16.

48. X １ and X ２ But both 【Transformation 38】 or alternatively, X １ but, 【Chemistry 39】 and X ２ but, 【Chemistry 40】 47. The compound of claim 46, wherein:

49. R １ and R ２ Both are C １０ -C １７ 46. ​​The compound of claim 45, or a pharmaceutically acceptable salt thereof, wherein the alkyl groups are branched and contain the same number of carbon atoms.

50. R １ But C ７ -C １１ Straight chain alkyl or C ９ -C １３ is a branched alkyl; R ２ But C １３ -C １９ branched alkyl; １ and R ２ At least one of them is C １３ 46. ​​The compound of claim 45, or a pharmaceutically acceptable salt thereof, which is not branched alkyl.

51. 46. ​​The compound of claim 45, wherein a and b are both 5, 6, 7, or 8, or a pharmaceutically acceptable salt thereof.

52. Bis(3-pentyloctyl)9-((2-oxaspiro[3.3]heptan-6-yl)amino)heptadecanedioate; Bis(3-pentyloctyl)9-((tetrahydro-2H-pyran-4-yl)amino)heptadecanedioate; Bis(3-pentyloctyl)9-(((tetrahydrofuran-3-yl)methyl)amino)heptadecanedioate; Bis(3-pentyloctyl)9-(((tetrahydro-2H-pyran-4-yl)methyl)amino)heptadecanedioate; and Bis(3-pentyloctyl)9-((oxetan-3-ylmethyl)amino)heptadecanedioate 2. The compound of claim 1, selected from:

53. 10. The compound of claim 9, or a pharmaceutically acceptable salt thereof, selected from compounds 6 to 57 as described herein.

54. 2. The compound of claim 1, which is bis(3-pentyloctyl) 9-((2-oxaspiro[3.3]heptan-6-yl)amino)heptadecanedioate or a pharmaceutically acceptable salt thereof.

55. 55. A lipid nanoparticle comprising a compound according to any one of claims 1 to 54, or a pharmaceutically acceptable salt thereof.

56. 56. The lipid nanoparticle of claim 55, further comprising at least one neutral lipid, at least one sterol, and at least one polymer-bound lipid.

57. 57. The lipid nanoparticle of claim 56, wherein the neutral lipid is selected from distearoylphosphatidylcholine (DSPC), dioleoylphosphatidylethanolamine (DOPE), dipalmitoylphosphatidylcholine (DPPC), dimyristoylphosphatidylcholine (DMPC) or a combination thereof.

58. The lipid nanoparticle of claim 56, wherein the sterol is cholesterol.

59. The lipid nanoparticle of claim 56, wherein the polymer-bound lipid is selected from DMPE-PEG2000, DPPE-PEG2000, DMG-PEG2000, DPG-PEG2000, PEG2000-c-DOMG, PEG-C-DOPG, or a combination thereof.

60. The lipid nanoparticle of claim 55, further comprising distearoylphosphatidylcholine (DSPC), cholesterol, and DMPE-PEG2000.

61. The lipid nanoparticle of claim 55, further comprising a nucleic acid segment.

62. The lipid nanoparticle of claim 61, wherein the nucleic acid segment is RNA.

63. The lipid nanoparticle of claim 61, wherein the nucleic acid segment is a modified mRNA.

64. A pharmaceutical composition comprising a plurality of lipid nanoparticles described in claim 61.

65. 65. The pharmaceutical composition of claim 64 for use in the treatment of a disease or disorder.

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