WO2025104695A1

WO2025104695A1 - Ionizable cationic lipids and lipid nanoparticles thereof

Info

Publication number: WO2025104695A1
Application number: PCT/IB2024/061417
Authority: WO
Inventors: Qi-Ying Hu
Original assignee: Corti Therapeutics Pte Ltd
Current assignee: Corti Therapeutics Pte Ltd
Priority date: 2023-11-16
Filing date: 2024-11-15
Publication date: 2025-05-22
Anticipated expiration: 2026-05-16

Abstract

Provided herein are ionizable lipids of the following structural formula: (I), or a pharmaceutically acceptable salt, solvate, or hydrate thereof, wherein values for the variables (e.g., G¹, G², L¹, L², M¹, M², R¹, R², R³, R⁴) are as described herein. Also provided are lipid nanoparticles comprising ionizable lipids of Formula (I), or a pharmaceutically acceptable salt, solvate, or hydrate thereof, and methods of using the ionizable lipids, e.g., to administer therapy to a subject in need thereof.

Description

Attorney Docket No. 01369-0001-00PCT IONIZABLE CATIONIC LIPIDS AND LIPID NANOPARTICLES THEREOF CROSS-REFERENCE TO RELATED APPLICATIONS [0001] This application claims the benefit of US Application No. 63/693,874, filed September 12, 2024, and US Application No. 63/599,781, filed November 16, 2023, the entire contents of which are incorporated by reference herein for all purposes. BACKGROUND [0002] The use of lipids to form lipid particles to deliver genetic material to a host has proven safe and effective. Regulators in the U.S and Europe approved the first therapeutic gene delivery via lipid nanoparticle in August of 2018. The therapy, patisiran, developed and marketed by Alnylam, consisted of nucleic acids wrapped in protective lipid shells. Two years later, the utility of lipid nanoparticles as nucleic acid delivery systems was further demonstrated with the approval of two mRNA-lipid nanoparticle vaccines from Moderna and Pfizer-BioNTech for the prevention of COVID-19. [0003] Cationic lipids are an important component of lipid nanoparticles. The cations present in the structure of cationic lipids aid in the encapsulation of negatively charged nucleic acids via ion pairing and also aid in the cellular uptake and subsequent escape of the nanoparticle. Ionizable cationic lipids such as MC3 (used in Onpattro) and 3D-P-DMA have demonstrated activity and tolerability in a variety of applications. However, they are not rapidly metabolized, and may take several weeks to clear from target tissues. [0004] Accordingly, there is a need for ionizable cationic lipids having novel or improved properties. SUMMARY [0005] The technology described herein relates to ionizable cationic lipids and lipid nanoparticles and/or pharmaceutical compositions comprising the ionizable cationic lipids. The technology can be used to administer therapeutic agents and/or nucleic acids to a subject, e.g., for the treatment or prevention of diseases or conditions, or to cells, e.g., cultured cells. [0006] Provided herein, in certain embodiments, are compounds of formula (I):

1 or a pharmaceutically acceptable salt, solvate, or hydrate thereof, wherein R¹, R², R³, R⁴, G¹, G², L¹, L², M¹, and M² are as defined herein. [0007] Also provided herein, in certain embodiments, is a lipid nanoparticle comprising a compound described herein and a therapeutic agent and/or nucleic acid. [0008] Also provided herein, in certain embodiments, is a pharmaceutical composition comprising a compound described herein, e.g., in the form of a lipid nanoparticle, including any of the lipid nanoparticles described herein, and a pharmaceutically acceptable carrier. [0009] Also provided herein, in certain embodiments, is a method of treating a disease or condition in a subject in need thereof, comprising administering to the subject a lipid nanoparticle described herein. [0010] Also provided herein, in certain embodiments, is a method of administering a therapeutic agent and/or nucleic acid to a subject, comprising administering to the subject a lipid nanoparticle comprising a compound of the present disclosure and the therapeutic agent and/or nucleic acid. [0011] Also provided herein, in certain embodiments, is use of the compounds, lipid nanoparticles, and pharmaceutical compositions described herein for a use described herein (e.g., for use in therapy, to administer a therapeutic agent and/or nucleic acid to a subject in need thereof, to treat a disease or condition in a subject in need thereof). Also provided herein, in certain embodiments, are compounds, lipid nanoparticles, and pharmaceutical compositions for a use described herein (e.g., for use in therapy, to administer a therapeutic agent and/or nucleic acid to a subject in need thereof, to treat a disease or condition in a subject in need thereof), wherein the compounds, lipid nanoparticles, and pharmaceutical compositions are described herein. [0012] Also provided herein, in certain embodiments, is use of a compound or lipid nanoparticle described herein for the preparation of a medicament, e.g., for use in therapy, to administer a therapeutic agent and/or nucleic acid to a subject in need thereof, to treat a disease or condition in a subject in need thereof. [0013] Also provided herein are the following numbered embodiments. [0014] Embodiment 1 is a compound of Formula I:

or a pharmaceutically acceptable salt, solvate, or hydrate thereof, wherein: L¹ and L² are each independently

, —O(C═O)—, —(C═O)O—, —C(=O)—, —O—, —S(O)x—, —S—S—, —NR^aC(═O)—, —C(═O)NR^a—, —NR^aC(═O)NR^a—, —OC(═O)NR^a—, —NR^aC(═O)O—, or a direct bond; each R^a is independently H or C1—C12 aliphatic; each R^b is independently C1—C8 aliphatic; each R^c is independently H or C₁—C₁₂ aliphatic; each x is independently 0, 1 or 2; M¹ and M² are each independently C1—C12 aliphatic or a direct bond; G¹ is C₁—C₂ alkylene or a direct bond; G² is —C(═O)S—, —C(═O)O—, —C(═O)—, —C(═O)N(H)—, —C(═O)N(C1—C12 aliphatic)—, —S(=O)2—, —S(=O)(=NH)—, —S(=O)(=N(C1—C12 aliphatic))—, —S(=O)—, —O—, —NH—, —CH2C(=O)—, —CH₂CF₂—, —C(=O)CF₂—, or a direct bond; R¹ and R² are each independently H or C1—C16 aliphatic; R³ is C4—C20 aliphatic optionally substituted with —L³⁰—R³⁰; L³⁰ is —O(C═O)—, —(C═O)O—, —C(=O)—, —O—, —S(O)_x3—, —S—S—, —NR^3aC(═O)—, —C(═O)NR^3a—, —NR^3aC(═O)NR^3a—, —OC(═O)NR^3a—, or —NR^3aC(═O)O—; each R^3a is independently H or C₁—C₁₂ aliphatic; R⁴ is

wherein indicates the point of attachment of R⁴ to the nitrogen atom of Formula I; G³ is a direct bond or C1—C12 alkylene; G⁴ is C₂—C₁₂ alkylene; R⁵ is a 4-, 5-, 6-, or 7-membered, carbon-linked, nitrogen-containing heterocycle, wherein each nitrogen atom of the heterocycle is independently substituted with R^d, and the heterocycle is optionally substituted on a carbon atom with R^e; R⁶ and R⁷ are each independently C₁—C₆ aliphatic, or R⁶ and R⁷, taken together with the nitrogen atom to which they are attached, form a 4-, 5-, 6-, or 7-membered, nitrogen- containing heterocycle, wherein each nitrogen atom of the heterocycle is independently substituted with R^d, and the heterocycle is optionally substituted on a carbon atom with R^e; each R^d is independently -(C1—C6 aliphatic) or –(C1—C6 aliphatic)OH; each R^e is independently -OH, -(C₁—C₆ aliphatic)OH, or -O(C₁—C₆ aliphatic); and provided: (i) when G² is —S(=O)₂—, either L¹ and L² are both a direct bond or neither L¹ nor L² is a direct bond; and (ii) when G² is a direct bond or —C(═O)—, R⁴ is

. [0015] Embodiment 2 is the compound of embodiment 1, which is a compound of Formula I-a:

, or a pharmaceutically acceptable salt, solvate, or hydrate thereof. [0016] Embodiment 3 is the compound of embodiment 1 or 2, wherein G² is —C(═O)S—, —C(═O)O—, —C(═O)—, —C(═O)N(H)—, —C(═O)N(C₁—C₁₂ aliphatic)—, —S(=O)₂—, —S(=O)—, or a direct bond. —, [0018] Embodiment 5 is the compound of any one of embodiments 1-4, wherein G² is —C(═O)S—, —C(═O)O—, or —S(=O)—. [0019] Embodiment 6 is the compound of any one of embodiments 1-4, wherein G² is —C(═O)S—, —S(=O)2—, or —S(=O)—. [0020] Embodiment 7 is the compound of any one of embodiments 1-5, wherein G² is —C(═O)S—. [0021] Embodiment 8 is the compound of embodiment 1 or 2, wherein G² is —C(═O)S—, —C(═O)O—, —C(═O)N(H)—, —C(═O)N(C1—C12 aliphatic)—, —S(=O)2—, —S(=O)(=NH)—, —S(=O)(=N(C₁—C₁₂ aliphatic))—,—S(=O)—, —O—, —NH—, —CH₂C(=O)—, —CH₂CF₂—, or —C(=O)CF₂—. [0022] Embodiment 9 is the compound of any one of embodiments 1, 2, and 8, wherein G² is —C(═O)S—, —S(=O)2—, —S(=O)(=NH)—, —S(=O)(=N(C1—C12 aliphatic))—,—S(=O)—, —O—, —CH₂C(=O)—, or —C(=O)CF₂—. [0023] Embodiment 10 is the compound of any one of embodiments 1-9, wherein R⁵ or the heterocycle formed by R⁶ and R⁷, taken together with the nitrogen atom to which they are attached, is azetidinyl, pyrrolidinyl, piperdinyl, azepanyl, morpholinyl, thiomorpholinyl, or piperazinyl. [0024] Embodiment 11 is the compound of any one of embodiments 1-10, wherein R⁵ is 3-azetidinyl, 2-pyrrolidinyl, 3-pyrrolidinyl, 2-piperdinyl, 3-piperdinyl, 4-piperdinyl, 3-azepanyl, 4-azepanyl, 2-morpholinyl, 2-thiomorpholinyl, or 2-piperazinyl. [0025] Embodiment 12 is the compound of any one of embodiments 1-11, wherein R⁵ is 2-morpholinyl, 3-piperdinyl, 2-pyrrolidinyl or 4-piperdinyl. [0026] Embodiment 13 is the compound of any one of embodiments 1-12, wherein L¹ and L² are each independently

—(C═O)O—, —C(=O)—, —O—, —S(O)_x—, —S—S—,—NR^aC(═O)—, —C(═O)NR^a—, —NR^aC(═O)NR^a—, —OC(═O)NR^a—, or —NR^aC(═O)O—. [0027] Embodiment 14 is the compound of any one of embodiments 1-13, wherein L¹ and L²

[0028] Embodiment 15 is the compound of any one of embodiments 1-14, wherein L¹ and L² are each independently

—(C═O)O—, or a direct bond. [0029] Embodiment 16 is the compound of any one of embodiments 1-15, wherein at least one

[0030] Embodiment 17 is the compound of embodiment any one of embodiments 1-16, wherein L¹ and L² are each independently —O(C═O)—, —(C═O)O—, or a direct bond. [0031] Embodiment 18 is the compound of any one of embodiments 1-17, which is a compound of formula I-b

, wherein m and n are each independently an integer from 0 to 5, and the sum of m and n is 2 to 5. [0032] Embodiment 19 is the compound of embodiment 18, wherein the sum of m and n is 4. [0033] Embodiment 20 is the compound of embodiment 18 or 19, wherein m and n are both 2. [0034] Embodiment 21 is the compound of any one of embodiments 1-20, wherein G³ is C1—C12 unbranched alkylene. [0035] Embodiment 22 is the compound of any one of embodiments 1-20, wherein G³ is C₁—C₁₂ branched alkylene. [0036] Embodiment 23 is the compound of any one of embodiments 1-22, wherein G³ is C₁—C₅ alkylene. [0039] Embodiment 26 is the compound of any one of embodiments 1-25, wherein R^c is C1—C12 aliphatic. [0040] Embodiment 27 is the compound of any one of embodiments 1-26, wherein R^c is C1—C12 unbranched aliphatic. [0041] Embodiment 28 is the compound of any one of embodiments 1-26, wherein R^c is C₁—C₁₂ branched aliphatic. [0042] Embodiment 29 is the compound of any one of embodiments 1-28, wherein R^c is C1—C12 alkyl. [0043] Embodiment 30 is the compound of any one of embodiments 1-29, wherein G¹ is a direct bond. [0044] Embodiment 31 is the compound of any one of embodiments 1-29, wherein G¹ is C1—C2 alkylene. [0045] Embodiment 32 is the compound of any one of embodiments 1-29, wherein M¹ and M² are the same. [0046] Embodiment 33 is the compound of any one of embodiments 1-32, wherein M¹ and M² are both a direct bond. [0047] Embodiment 34 is the compound of any one of embodiments 1-32, wherein M¹ and M² are both C1—C12 aliphatic. [0048] Embodiment 35 is the compound of any one of embodiments 1-32 and 34, wherein M¹ and M² are both C₁—C₁₂ branched aliphatic. [0049] Embodiment 36 is the compound of any one of embodiments 1-32 and 34, wherein M¹ and M² are both C1—C12 unbranched aliphatic. [0050] Embodiment 37 is the compound of any one of embodiments 1-32 and 34-36 wherein M¹ and M² are both C6—C12 aliphatic. [0051] Embodiment 38 is the compound of any one of embodiments 1-32, wherein M¹ and M² are each independently C₁—C₁₂ unbranched aliphatic or a direct bond. [0052] Embodiment 39 is the compound of any one of embodiments 1-32, wherein M¹ and M² are each independently C1—C12 branched aliphatic or a direct bond. [0053] Embodiment 40 is the compound of any one of embodiments 1-32, 38 and 39, wherein M¹ and M² are each independently C₁—C₁₂ alkylene or a direct bond. nd R² nd R² [0056] Embodiment 43 is the compound of any one of embodiments 1-41, wherein R¹ and R² are both C1—C16 unbranched aliphatic. [0057] Embodiment 44 is the compound of any one of embodiments 1-43, wherein R¹ and R² are C1—C16 alkyl. [0058] Embodiment 45 is the compound of any one of embodiments 1-40, wherein R¹ and R² are each independently H or C₁—C₁₂ aliphatic. [0059] Embodiment 46 is the compound of any one of embodiments 1-45, wherein R¹ and R² are the same. [0060] Embodiment 47 is the compound of any one of embodiments 1-40, 45 and 46, wherein R¹ and R² are both H. [0061] Embodiment 48 is the compound of any one of embodiments 1-45, wherein R¹ and R² are different. [0062] Embodiment 49 is the compound of any one of embodiments 1-48, wherein R³ is C4—C20 aliphatic. [0063] Embodiment 50 is the compound of any one of embodiments 1-48, wherein R³ is C₄—C₂₀ unbranched aliphatic. [0064] Embodiment 51 is the compound of any one of embodiments 1-49, wherein R³ is C4—C20 branched aliphatic. [0065] Embodiment 52 is the compound of any one of embodiments 1-50, wherein R³ is C₄—C₂₀ alkyl. [0066] Embodiment 53 is the compound of any one of embodiments 1-51, wherein R³ is C4—C10 aliphatic. [0067] Embodiment 54 is the compound of any one of embodiments 1, 3-9, 13-17, and 24-53 wherein

. [0068] Embodiment 55 is the compound of any one of embodiments 1, 3-9, 13-17, and 24-54, wherein G⁴ is C2—C12 unbranched alkylene. [0069] Embodiment 56 is the compound of any one of embodiments 1, 3-9, 13-17, and 24-54, wherein G⁴ is C₂—C₁₂ branched alkylene. [0070] Embodiment 57 is the compound of any one of embodiments 1 3-9 13-17 and 24-56, 4-57, [0072] Embodiment 59 is the compound of any one of embodiments 1, 3-9, 13-17, and 24-58, wherein R⁶ and R⁷ are each independently C1—C6 alkyl. [0073] Embodiment 60 is the compound of any one of embodiments 1, 3-9, 13-17, and 24-59, wherein R⁶ and R⁷ are each independently C1—C3 alkyl. [0074] Embodiment 61 is the compound of any one of embodiments 1, 3-9, 13-17, and 24-60, wherein R⁶ and R⁷ are the same. [0075] Embodiment 62 is the compound of any one of embodiments 1, 3-9, 13-17, and 24-61, wherein R⁶ and R⁷ are both methyl. [0076] Embodiment 63 is the compound of any one of embodiments 1, 3-9, 13-17, and 24-60, wherein R⁶ and R⁷ are different. [0077] Embodiment 64 is the compound of any one of embodiments 1, 3-9, 13-17, and 24-57, wherein R⁶ and R⁷ taken together with the nitrogen atom to which they are attached, form a 4-, 5-, 6-, or 7-membered, nitrogen-containing heterocycle, wherein each nitrogen atom of the heterocycle is independently substituted with R^d, and the heterocycle is optionally substituted on a carbon atom with R^e. [0078] Embodiment 65 is a compound selected from: ,

, ,

,

, or a pharmaceutically acceptable salt, solvate, or hydrate thereof. [0079] Embodiment 66 is a lipid nanoparticle comprising the compound of any one of embodiments 1-65 and a therapeutic agent. [0080] Embodiment 67 is a lipid nanoparticle comprising the compound of any one of embodiments 1-65 and one or more nucleic acids. [0081] Embodiment 68 is the lipid nanoparticle of embodiment 66 or 67, further comprising a steroid. [0082] Embodiment 69 is the lipid nanoparticle of any one of embodiments 66-68, further comprising a PEG lipid. [0083] Embodiment 70 is the lipid nanoparticle of any one of embodiments 66-69, further comprising a phospholipid. [0084] Embodiment 71 is a lipid nanoparticle comprising the compound of any one of embodiments 1-65, a steroid, a PEG lipid, and a phospholipid, optionally further comprising a therapeutic agent or one or more nucleic acids. [0085] Embodiment 72 is the lipid nanoparticle of any one of embodiments 66-70, wherein one or more of the nucleic acids is a therapeutic agent. [0086] Embodiment 73 is the lipid nanoparticle of any one of embodiments 67-72, wherein at least one of the one or more nucleic acids is DNA. [0087] Embodiment 74 is the lipid nanoparticle of any one of embodiments 67-72, wherein at least one of the one or more nucleic acids is RNA. [0088] Embodiment 75 is the lipid nanoparticle of any one of embodiments 67-74, wherein at least one of the one or more nucleic acids comprises a modified nucleotide. [0089] Embodiment 76 is a pharmaceutical composition comprising the compound of any one of embodiments 1-65 or lipid nanoparticle of any one of embodiments 66-75, and a pharmaceutically acceptable carrier. [0090] Embodiment 77 is a method of treating a disease or condition in a subject in need thereof, comprising administering to the subject the lipid nanoparticle of any one of embodiments 66-75. [0091] Embodiment 78 is a method of administering a therapeutic agent or nucleic acid to a subject, comprising administering to the subject a lipid nanoparticle of any one of embodiments 66-75. [0092] Embodiment 79 is use of a lipid nanoparticle of any one of embodiments 66-75 for administering a therapeutic agent or nucleic acid to a subject in need thereof. [0093] Embodiment 80 is use of a lipid nanoparticle of any one of embodiments 66-75 to treat a disease or condition in a subject in need thereof. [0094] Embodiment 81 is the lipid nanoparticle of any one of embodiments 66-75 for administering a therapeutic agent or nucleic acid to a subject in need thereof. [0095] Embodiment 82 is the method, use, or lipid nanoparticle of any one of embodiments 77-81, wherein the subject is a mammal, optionally wherein the subject is a human. [0096] Embodiment 83 is use of the lipid nanoparticle of any one of embodiments 66-75 for preparation of a medicament. [0097] Embodiment 84 is the lipid nanoparticle of any one of embodiments 66-75 for use in therapy. DETAILED DESCRIPTION Definitions [0098] Compounds described herein include those described generally, and are further illustrated by the classes, subclasses, and species disclosed herein. As used herein, the following definitions shall apply unless otherwise indicated. For purposes of this invention, the chemical elements are identified in accordance with the Periodic Table of the Elements, CAS version, Handbook of Chemistry and Physics, 75^th Ed. Additionally, general principles of organic chemistry are described in “Organic Chemistry”, Thomas Sorrell, University Science Books, Sausalito: 1999, and “March’s Advanced Organic Chemistry”, 5^th Ed.: Smith, M.B. and March, J., John Wiley & Sons, New York: 2001, the relevant contents of which are incorporated herein by reference. [0099] Unless specified otherwise within this specification, the nomenclature used in this specification generally follows the examples and rules stated in Nomenclature of Organic Chemistry, Sections A, B, C, D, E, F, and H, Pergamon Press, Oxford, 1979, which is incorporated by reference herein for its chemical structure names and rules on naming chemical structures. Optionally, a name of a compound may be generated using a chemical naming program (e.g., CHEMDRAW®, version 17.0.0.206, PerkinElmer Informatics, Inc.). [00100] Section headings are provided herein solely for the convenience of the reader and do not limit the disclosure. [00101] When introducing elements disclosed herein, the articles “a,” “an,” “the,” and “said” are intended to mean that there are one or more of the elements unless the context requires otherwise. Further, the one or more elements may be the same or different. [00102] “Or” is used in the inclusive sense, i.e., equivalent to “and/or”, unless the context requires otherwise. [00103] “About” means within an acceptable error range for the particular value, as determined by one of ordinary skill in the art. Typically, an acceptable error range for a particular value depends, at least in part, on how the value is measured or determined, e.g., the limitations of the measurement system. For example, “about” can mean within an acceptable standard deviation, per the practice in the art. Alternatively, “about” can mean a range of ± 20%, e.g., ± 10%, ± 5% or ± 1% of a given value. It is to be understood that the term “about” can precede any particular value specified herein, except for particular values used in the Examples. [00104] "Aliphatic” refers to a saturated or unsaturated, branched or straight-chain, monovalent, hydrocarbon radical having the specified number of carbon atoms. Thus, “C1-C8 aliphatic” refers to a radical that is saturated or unsaturated having from 1-8 carbon atoms in a branched or linear arrangement. In some aspects, aliphatic is C1-C30 aliphatic, e.g., C5-C30 aliphatic, C₁-C₂₅ aliphatic, C₅-C₂₅ aliphatic, C₁₀-C₂₅ aliphatic, C₁₅-C₂₅ aliphatic, C₄-C₂₀ aliphatic, C10-C20 aliphatic, C15-C20 aliphatic, C1-C15 aliphatic, C1-C12 aliphatic, C1-C8 aliphatic, C1-C4 aliphatic, or C1-C2 aliphatic. In some aspects, aliphatic is branched. In some aspects, aliphatic is unbranched. Examples of aliphatic include alkyl, alkylene, alkenyl, alkynyl, cycloalkyl, cycloalkenyl, and cycloalkynyl. In some aspects, aliphatic is saturated, e.g., as in alkyl. [00105] “Alkyl” refers to a saturated, branched or straight-chain, monovalent, hydrocarbon radical having the specified number of carbon atoms. Thus, “C₁-C₈ alkyl” refers to a saturated monoradical having from 1-8 carbon atoms in a branched or linear arrangement. In some aspects, alkyl is C1-C30 alkyl, e.g., C5-C30 alkyl, C1-C25 alkyl, C5-C25 alkyl, C10-C25 alkyl, C15- C25 alkyl, C4-C20 alkyl, C10-C20 alkyl, C15-C20 alkyl, C1-C15 alkyl, C1-C12 alkyl, C1-C8 alkyl, C1- C₄ alkyl, or C₁-C₂ alkyl. Examples of alkyl groups include methyl, ethyl, n‐propyl, isopropyl, n‐ butyl, isobutyl, sec‐butyl, t‐butyl, n‐pentyl, isopentyl, neopentyl, 2‐methylpentyl, n‐hexyl, and the like. [00106] “Alkylene” refers to a saturated, branched or straight-chain, divalent, hydrocarbon radical having the specified number of carbon atoms. Thus, “C1-C8 alkylene” refers to a saturated diradical having from 1-8 carbon atoms in a branched or linear arrangement. In some aspects, alkylene is C₁-C₃₀ alkylene, e.g., C₅-C₃₀ alkylene, C₁-C₂₅ alkylene, C₅-C₂₅ alkylene, C₁₀- C₂₅ alkylene, C₁₅-C₂₅ alkylene, C₄-C₂₀ alkylene, C₁₀-C₂₀ alkylene, C₁₅-C₂₀ alkylene, C₁-C₁₅ alkylene, C1-C12 alkylene, C1-C8 alkylene, C1-C4 alkylene, or C1-C2 alkylene. Examples of alkylene groups include methylene, ethylene, n‐propylene, isopropylene, n‐butylene, isobutylene, sec‐butylene, t‐butylene, n‐pentylene, isopentylene, neopentylene, 2‐ methylpentylene, n‐hexylene, and the like. [00107] “Alkenyl” refers to a branched or straight-chain, monovalent, hydrocarbon monoradical having at least one carbon-carbon double bond and the specified number of carbon atoms. Thus, “(C2-C8)alkenyl” refers to a radical having at least one carbon-carbon double bond and from 2-8 carbon atoms in a branched or linear arrangement. In some aspects, alkenyl is C1- C₃₀ alkenyl, e.g., C₅-C₃₀ alkenyl, C₁-C₂₅ alkenyl, C₅-C₂₅ alkenyl, C₁₀-C₂₅ alkenyl, C₁₅-C₂₅ alkenyl, C₄-C₂₀ alkenyl, C₁₀-C₂₀ alkenyl, C₁₅-C₂₀ alkenyl, C₁-C₁₅ alkenyl, C₁-C₁₂ alkenyl, C₁-C₈ alkenyl, C1-C4 alkenyl, or C1-C2 alkenyl. Examples of alkenyl groups include ethenyl, 2‐ propenyl, 1‐propenyl, 2‐methyl‐1‐propenyl, 1‐butenyl, 2‐butenyl, 1‐pentenyl, 2‐pentenyl, 3‐ pentenyl, allyl, 1, 3‐butadienyl, 1, 3‐dipentenyl, 1,4-dipentenyl, 1‐hexenyl, 1,3‐hexenyl, 1,4‐ hexenyl, 1,3,5‐trihexenyl, 2,4‐dihexenyl, and the like. [00108] “Heterocyclyl,” “heterocycle,” or “heterocyclic ring” refers to a stable non-aromatic ring system radical having the specified number of ring atoms, wherein the ring atoms consist of carbon atoms and one or more (e.g., one, two, three, four, five, etc.; one to three; one or two) heteroatoms selected from nitrogen, oxygen and sulfur. Unless stated otherwise specifically in the specification, heterocyclyl may be monocyclic or polycyclic (e.g., bicyclic, tricyclic, or tetracyclic), and may include fused or bridged ring systems; and nitrogen, carbon or sulfur atoms in the heterocyclyl radical may be optionally oxidized; nitrogen atoms may be optionally quaternized. A heterocyclyl may be partially or fully saturated. In some aspects, heterocyclyl is saturated. In some aspects, heterocyclyl is monocyclic (e.g., monocyclic and saturated). Examples of heterocyclyl include, but are not limited to, azetidinyl, pyrrolidinyl, piperdinyl, azepanyl, morpholinyl, thiomorpholinyl, and piperazinyl. Additional examples of heterocyclyl include, but are not limited to, dioxolanyl, thienyl[1,3]dithianyl, decahydroisoquinolyl, imidazolinyl, imidazolidinyl, isothiazolidinyl, isoxazolidinyl, octahydroindolyl, octahydroisoindolyl, 2-oxopiperazinyl, 2-oxopiperidinyl, 2-oxopyrrolidinyl, oxazolidinyl, 4- piperidonyl, pyrazolidinyl, quinuclidinyl, thiazolidinyl, tetrahydrofuryl, trithianyl, tetrahydropyranyl, thiamorpholinyl, 1-oxothiomorpholinyl, and 1,1-dioxothiomorpholinyl. [00109] A “nitrogen-containing heterocycle” refers to a heterocyclyl, as that term is described herein, wherein at least one of the heteroatoms is a nitrogen. When a heterocycle is referred to herein as “nitrogen-linked,” the atom of the heterocycle acting as the point of attachment for the heterocyclyl radical is a nitrogen atom. When a heterocycle is referred to herein as “carbon- linked,” the atom of the heterocycle acting as the point of attachment for the heterocyclyl radical is a carbon atom. [00110] The term “substituted” refers to replacement of a hydrogen atom with a suitable substituent. Typically, the suitable substituent replaces a hydrogen atom bound to a carbon atom, but a substituent may also replace a hydrogen bound to a heteroatom, such as a nitrogen, oxygen, or sulfur atom. It will be understood that “substitution” or “substituted with” includes the implicit proviso that such substitution is in accordance with permitted valence of the substituted atom. It is also preferred that the substituent, and the substitution, result in a stable compound, e.g., which does not spontaneously undergo transformation such as by rearrangement, cyclization, elimination, etc. [00111] The term “optionally substituted”, as used herein, means that substitution is optional and, therefore, it is possible for the atom or moiety designated as “optionally substituted” to be unsubstituted or substituted. In some aspects, an optionally substituted group is unsubstituted. In some aspects, an optionally substituted group is substituted. Unless otherwise indicated, e.g., as with the terms “substituted” or “optionally substituted,” a group designated herein is unsubstituted. [00112] As used herein, the term “compound of the disclosure” refers to a compound of any of the structural formulas depicted herein (e.g., a compound of Formula (I), an exemplified compound), as well as isomers, such as stereoisomers (including diastereoisomers, enantiomers and racemates) and tautomers, thereof, isotopically labeled variants thereof (including those with deuterium substitutions), and inherently formed moieties (e.g., polymorphs and/or solvates, such as hydrates) thereof. When a moiety is present that is capable of forming a salt, then salts are included as well, in particular, pharmaceutically acceptable salts thereof. [00113] When the phrase “the nitrogen atom of Formula I” is used herein, such phrase refers to the nitrogen atom in Formula I below marked with an asterisk (*) (or its corresponding atom in any of the structural formulas depicted herein, such as an exemplified compound):

It will be understood that the term is not meant to exclude presence of additional nitrogen atoms in the referenced compound. [00114] Compounds of the disclosure may have asymmetric centers, chiral axes, and chiral planes (e.g., as described in: E. L. Eliel and S. H. Wilen, Stereo-chemistry of Carbon Compounds, John Wiley & Sons, New York, 1994, pages 1119-1190), and occur as racemic mixtures, individual isomers (e.g., diastereomers, enantiomers, geometrical isomers (including cis and trans double bond isomers), conformational isomers (including rotamers and atropisomers), and tautomers) and intermediate mixtures, with all possible isomers and mixtures thereof being included, unless otherwise indicated. [00115] When a disclosed compound is depicted by structure without indicating the stereochemistry, and the compound has one or more chiral centers, it is to be understood that the structure encompasses one enantiomer or diastereomer of the compound separated or substantially separated from the corresponding optical isomer(s), a racemic mixture of the compound and mixtures enriched in one enantiomer or diastereomer relative to its corresponding optical isomer(s). When a disclosed compound is depicted by a structure indicating stereochemistry, and the compound has one or more chiral centers, the stereochemistry indicates absolute configuration of the substituents around the one or more chiral centers. “R” and “S” can also or alternatively be used to indicate the absolute configuration of substituents around one or more chiral carbon atoms. D- and L- can also or alternatively be used to designate stereochemistry. [00116] “Enantiomers” are pairs of stereoisomers that are non-superimposable mirror images of one another, most commonly because they contain an asymmetrically substituted carbon atom that acts as a chiral center. [00117] “Diastereomers” are non-superimposable stereoisomers that are not related as mirror images, most commonly because they contain two or more asymmetrically substituted carbon atoms. [00118] “Racemate” or “racemic mixture,” as used herein, refers to a mixture containing equimolar quantities of two enantiomers of a compound. Such mixtures exhibit no optical activity (i.e., they do not rotate a plane of polarized light). [00119] Percent enantiomeric excess (ee) is defined as the absolute difference between the mole fraction of each enantiomer multiplied by 100% and can be represented by the following _{equation: ee =}

_{× 100%, where R and S represent the respective fractions of each} enantiomer in a mixture, such that R + S = 1. An enantiomer may be present in an ee of at least or about 50%, about 60%, about 70%, about 80%, about 90%, about 95%, about 98%, about 99% or about 99.9%. [00120] Percent diastereomeric excess (de) is defined as the absolute difference between the mole fraction of each diastereomer multiplied by 100% and can be represented by the following _{equation: de =}

_{where D1 and (D2 + D3 + D4…) represent the} respective fractions of each diastereomer in a mixture, such that D1 + (D2 + D3 + D4…) = 1. A diastereomer may be present in a de of at least or about 50%, about 60%, about 70%, about 80%, about 90%, about 95%, about 98%, about 99% or about 99.9%. [00121] Unless otherwise stated, compounds of the disclosure include compounds that differ only in the presence of one or more isotopically enriched atoms. For example, compounds produced by the replacement of a hydrogen with deuterium or tritium, or of a carbon with a ¹³C- or ¹⁴C-enriched carbon are within the scope of this invention. In all provided structures, any hydrogen atom can also be independently selected from deuterium (²H) and/or tritium (³H). Such compounds are useful, for example, as analytical tools, as probes in biological assays, or in accordance with the present invention. [00122] The phrase “pharmaceutically acceptable” means that the substance or composition the phrase modifies is, within the scope of sound medical judgment, suitable for use in contact with the tissues of humans and lower animals without undue toxicity, irritation, allergic response and the like, and are commensurate with a reasonable benefit/risk ratio. [00123] As used herein, the term “pharmaceutically acceptable salt” refers to those salts which are, within the scope of sound medical judgment, suitable for use in contact with the tissues of mammals without undue toxicity, irritation, allergic response and the like, and are commensurate with a reasonable benefit/risk ratio. Pharmaceutically acceptable salts are well known in the art. For example, S. M. Berge et al., describe pharmaceutically acceptable salts in detail in J. Pharmaceutical Sciences, 1977, 66, 1–19, the relevant teachings of which is incorporated herein by reference in its entirety. Pharmaceutically acceptable salts of the compounds described herein include salts derived from suitable inorganic and organic acids, and suitable inorganic and organic bases. [00124] Examples of salts derived from suitable acids include salts of an amino group formed with inorganic acids such as hydrochloric acid, hydrobromic acid, phosphoric acid, sulfuric acid and perchloric acid, or with organic acids such as acetic acid, oxalic acid, maleic acid, tartaric acid, citric acid, succinic acid or malonic acid or by using other methods used in the art, such as ion exchange. Other pharmaceutically acceptable salts derived from suitable acids include adipate, alginate, ascorbate, aspartate, benzenesulfonate, benzoate, bisulfate, borate, butyrate, camphorate, camphorsulfonate, cinnamate, citrate, cyclopentanepropionate, digluconate, dodecylsulfate, ethanesulfonate, formate, fumarate, glucoheptonate, glycerophosphate, gluconate, glutarate, glycolate, hemisulfate, heptanoate, hexanoate, hydroiodide, hydroxybenzoate, 2-hydroxy-ethanesulfonate, hydroxymaleate, lactobionate, lactate, laurate, lauryl sulfate, malate, maleate, malonate, methanesulfonate, 2-naphthalenesulfonate, nicotinate, nitrate, oleate, oxalate, palmitate, pamoate, pectinate, persulfate, 2-phenoxybenzoate, phenylacetate, 3-phenylpropionate, phosphate, pivalate, propionate, pyruvate, salicylate, stearate, succinate, sulfate, tartrate, thiocyanate, p-toluenesulfonate, undecanoate, valerate salts, and the like. [00125] Either the mono-, di- or tri-acid salts can be formed, and such salts can exist in either a hydrated, solvated or substantially anhydrous form. [00126] Salts derived from appropriate bases include salts derived from inorganic bases, such as alkali metal, alkaline earth metal, and ammonium bases, and salts derived from aliphatic, alicyclic or aromatic organic amines, such as methylamine, trimethylamine and picoline, or N⁺((C1-C4)alkyl)4 salts. Representative alkali or alkaline earth metal salts include sodium, lithium, potassium, calcium, magnesium, barium and the like. Further pharmaceutically acceptable salts include, when appropriate, nontoxic ammonium, quaternary ammonium, and amine cations formed using counterions such as halide, hydroxide, carboxyl, sulfate, phosphate, nitrate, lower alkyl sulfonate and aryl sulfonate. [00127] Compounds described herein can also exist as “solvates” or “hydrates.” A “hydrate” is a compound that exists in a composition with one or more water molecules. A hydrate can include water in stoichiometric quantities, such as a monohydrate or a dihydrate, or can include water in random amounts. A “solvate” is similar to a hydrate, except that a solvent other than water, such as methanol, ethanol, dimethylformamide, diethyl ether, or the like replaces water. Mixtures of such solvates or hydrates can also be prepared. The source of such solvate or hydrate can be from the solvent of crystallization, inherent in the solvent of preparation or crystallization, or adventitious to such solvent. [00128] Use herein, for example, in the claims, of the phrase, “a pharmaceutically acceptable salt, solvate, or hydrate thereof,” will be understood to include pharmaceutically acceptable salts, solvates, or hydrates of the reference compound, or any combination thereof, such as a solvated pharmaceutically acceptable salt of the reference compound. [00129] “Pharmaceutically acceptable carrier” refers to a non-toxic carrier or excipient that does not destroy the pharmacological activity of the agent with which it is formulated and is nontoxic when administered in doses sufficient to deliver a therapeutic amount of the agent. Pharmaceutically acceptable carriers that may be used in the compositions described herein include, but are not limited to, ion exchangers, alumina, aluminum stearate, lecithin, serum proteins, such as human serum albumin, buffer substances such as phosphates, glycine, sorbic acid, potassium sorbate, partial glyceride mixtures of saturated vegetable fatty acids, water, salts or electrolytes, such as protamine sulfate, disodium hydrogen phosphate, potassium hydrogen phosphate, sodium chloride, zinc salts, colloidal silica, magnesium trisilicate, polyvinyl pyrrolidone, cellulose-based substances, polyethylene glycol, sodium carboxymethylcellulose, polyacrylates, waxes, polyethylene-polyoxypropylene-block polymers, polyethylene glycol and wool fat. [00130] “Treating,” as used herein, refers to taking steps to deliver a therapy to a subject, such as a mammal, in need thereof (e.g., as by administering to a subject one or more therapeutic agents). “Treating” includes inhibiting a disease or condition (e.g., as by slowing or stopping its progression or causing regression of the disease or condition) and relieving symptoms resulting from a disease or condition. “Treating” also includes preventing a disease or condition in a subject (e.g., as by administering to a subject one or more therapeutic agents before the onset of a disease or condition) such that onset of the disease or condition is delayed or prevented, or symptoms of the disease or condition, upon onset, are less severe. The various compounds, compositions, and/or therapeutic agents described herein may be administered for treating prophylactically (to prevent disease or condition) or therapeutically (e.g., to subjects suffering from a disease or condition). Such subjects may be identified using standard clinical methods. [00131] “A therapeutically effective amount” is an amount effective, at dosages and for periods of time necessary, to achieve a desired therapeutic result. The full therapeutic effect does not necessarily occur by administration of one dose and may occur only after administration of a series of doses. Thus, a therapeutically effective amount may be administered in one or more administrations. A therapeutically effective amount may vary according to factors such as disease state, age, sex, and weight of a mammal, mode of administration and the ability of a therapeutic, or combination of therapeutics, to elicit a desired response in an individual. [00132] As used herein, “subject” includes humans, domestic animals, such as laboratory animals (e.g., dogs, monkeys, pigs, rats, mice, etc.), household pets (e.g., cats, dogs, rabbits, etc.) and livestock (e.g., pigs, cattle, sheep, goats, horses, etc.), and non-domestic animals. In some aspects, a subject is a human. [00133] Unless otherwise defined, the technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which this application belongs. Ionizable Cationic Lipids [00134] Typically, ionizable lipids or ionizable cationic lipids are lipids that are designed to be uncharged at standard physiological pH (~7) but acquire a positive charge at lower (more acidic) pH. Ionizable cationic lipids are thus often able to undergo ionization or de-ionization with changes in the environmental pH, as for example, by protonation or deprotonation of a nitrogen atom. [00135] Provided herein, in certain embodiments, are compounds of formula (I):

, —O(C═O)—, —(C═O)O—, —C(=O)—, —O—, —S(O)x—, —S—S—,—NR^aC(═O)—, —C(═O)NR^a—, —NR^aC(═O)NR^a—, —OC(═O)NR^a—, —NR^aC(═O)O—, or a direct bond; each R^a is independently H or C₁—C₁₂ aliphatic; each R^b is independently C1—C8 aliphatic; each R^c is independently H or C1—C12 aliphatic; each x is independently 0, 1 or 2; M¹ and M² are each independently C1—C12 aliphatic or a direct bond; G¹ is C1—C2 alkylene or a direct bond; G² is —C(═O)S—, —C(═O)O—, —C(═O)—, —C(═O)N(H)—, —C(═O)N(C₁—C₁₂ aliphatic)—, —S(=O)₂—, —S(=O)(=NH)—, —S(=O)(=N(C₁—C₁₂ aliphatic))—, —S(=O)—, —O—, —NH—, —CH2C(=O)O—, —CH2CF2—, —C(=O)CF2— , or a direct bond; R¹ and R² are each independently H or C₁—C₁₆ aliphatic; R³ is C4—C20 aliphatic optionally substituted with —L³⁰—R³⁰; L³⁰ is —O(C═O)—, —(C═O)O—, —C(=O)—, —O—, —S(O)x3—, —S—S—, —NR^3aC(═O)—, —C(═O)NR^3a—, —NR^3aC(═O)NR^3a—, —OC(═O)NR^3a—, or —NR^3aC(═O)O—; each R^3a is independently H or C1—C12 aliphatic; R³⁰ is H or C₁—C₁₂ aliphatic; x3 is 0, 1, or 2; R⁴ is or , wherein indicates the point of attachment of R⁴ to the nitrogen atom of Formula I; G³ is a direct bond or C1—C12 alkylene; G⁴ is C₂—C₁₂ alkylene; R⁵ is a 4-, 5-, 6-, or 7-membered, carbon-linked, nitrogen-containing heterocycle, wherein each nitrogen atom of the heterocycle is independently substituted with R^d, and the heterocycle is optionally substituted on a carbon atom with R^e; each R^d is independently -(C₁—C₆ aliphatic) or –(C₁—C₆ aliphatic)OH; each R^e is independently -OH, -(C1—C6 aliphatic)OH, or -O(C1—C6 aliphatic); and R⁶ and R⁷ are each independently C1—C6 aliphatic, or R⁶ and R⁷, taken together with the nitrogen atom to which they are attached, form a 4-, 5-, 6-, or 7-membered, nitrogen- containing heterocycle, wherein each nitrogen atom of the heterocycle is independently substituted with R^d, and the heterocycle is optionally substituted on a carbon atom with R^e; provided: (i) when G² is —S(=O)₂—, either L¹ and L² are both a direct bond or neither L¹ nor L² is a direct bond; and (ii) when G² is a direct bond or —C(═O)—, R⁴ is

. [00136] In some embodiments is a compound of Formula I-a:

, or a pharmaceutically acceptable salt, solvate, or hydrate thereof, wherein values for the remaining variables (e.g., R¹, R², R³, R⁵, L¹, L², M¹, M², G¹, G², G³) are as described with respect to a compound of Formula I or elsewhere herein. [00137] Yet another embodiment is a compound of Formula I-b:

or a pharmaceutically acceptable salt, solvate, or hydrate thereof, wherein m and n are each independently an integer from 0 to 5, and the sum of m and n is 2 to 5. Values for the remaining variables (e.g., R¹, R², R³, L¹, L², M¹, M², G¹, G³, R^d, R^e) are as described with respect to a compound of Formula I or elsewhere herein. [00138] In some embodiments, G² is —C(═O)S—, —C(═O)O—, —C(═O)—, —C(═O)N(H)—, —C(═O)N(C1—C12 aliphatic)—, —S(=O)2—, —S(=O)(=NH)—, —S(=O)(=N(C₁—C₁₂ aliphatic))—, —S(=O)—, —O—, —NH—, —CH₂C(=O)O—, —CH2CF2—, or —C(=O)CF2—. In some embodiments, G² is —C(═O)S—, —C(═O)O—, —C(═O)N(H)—, —C(═O)N(C1—C12 aliphatic)—, —S(=O)2—, —S(=O)(=NH)—, —S(=O)(=N(C1—C12 aliphatic))—, —O—, —CH2C(=O)O—, —C(=O)CF2—, or —S(=O)—. In some embodiments, G² is —C(═O)S—, —S(=O)₂—, —S(=O)(=NH)—, —S(=O)(=N(C1—C12 aliphatic))—, or —S(=O)—. In some embodiments, G² is —S(=O)(=NH)—. In some embodiments, G² is —S(=O)(=N(C1—C12 aliphatic))— (e.g., —S(=O)(=N(C₁—C₂ aliphatic)). In some embodiments, G² is —O—, —NH—, —CH2C(=O)O—, or —CH2CF2—. [00139] In some embodiments, G² is —C(═O)S—, —C(═O)O—, —C(═O)—, —C(═O)N(H)—, —C(═O)N(C₁—C₁₂ aliphatic)—, —S(=O)₂—, —S(=O)—, or a direct bond. In some embodiments, G² is —C(═O)S—, —C(═O)O—, —C(═O)N(H)—, —C(═O)N(C1—C12 aliphatic)—, —S(=O)2—, or —S(=O)—. In some embodiments, G² is —C(═O)S—, —C(═O)O—, or —S(=O)—. In some embodiments, —C(═O)S—, —S(=O)2—, or —S(=O)—. In some embodiments, G² is —C(═O)S— or —S(=O)—. In some embodiments, G² is —C(═O)S—. In some embodiments, G² is —C(═O)O—. In some embodiments, G² is —C(═O)—. In some embodiments, G² is —C(═O)N(H)—. In some embodiments, G² is —C(═O)N(C₁—C₁₂ aliphatic)—. In some embodiments, G² is —S(=O)₂—. In some embodiments, G² is —S(=O)—. In some embodiments, G² is a direct bond. [00140] In some embodiments, R⁵ or the heterocycle formed by R⁶ and R⁷, taken together with the nitrogen atom to which they are attached, is azetidinyl, pyrrolidinyl, piperdinyl, azepanyl, morpholinyl, thiomorpholinyl, or piperazinyl. In some embodiments, R⁵ or the heterocycle formed by R⁶ and R⁷, taken together with the nitrogen atom to which they are attached, is azetidinyl. In some embodiments, R⁵ or the heterocycle formed by R⁶ and R⁷, taken together with the nitrogen atom to which they are attached, is pyrrolidinyl. In some embodiments, R⁵ or the heterocycle formed by R⁶ and R⁷, taken together with the nitrogen atom to which they are attached, is piperdinyl. In some embodiments, R⁵ or the heterocycle formed by R⁶ and R⁷, taken together with the nitrogen atom to which they are attached, is azepanyl. In some embodiments, R⁵ or the heterocycle formed by R⁶ and R⁷, taken together with the nitrogen atom to which they are attached, is morpholinyl. In some embodiments, R⁵ or the heterocycle formed by R⁶ and R⁷, taken together with the nitrogen atom to which they are attached, is thiomorpholinyl. In some embodiments, R⁵ or the heterocycle formed by R⁶ and R⁷, taken together with the nitrogen atom to which they are attached, is piperazinyl. [00141] In some embodiments, R⁵ is 3-azetidinyl, 2-pyrrolidinyl, 3-pyrrolidinyl, 2-piperdinyl, 3-piperdinyl, 4-piperdinyl, 3-azepanyl, 4-azepanyl, 2-morpholinyl, 2-thiomorpholinyl, or 2- piperazinyl. In some embodiments, R⁵ is 2-morpholinyl, 3-piperdinyl, 2-pyrrolidinyl or 4- piperdinyl. In some embodiments, R⁵ is 3-azetidinyl. In some embodiments, R⁵ is 2-pyrrolidinyl. In some embodiments, R⁵ is 3-pyrrolidinyl. In some embodiments, R⁵ is 2-piperdinyl. In some embodiments, R⁵ is 3-piperdinyl. In some embodiments, R⁵ is 4-piperdinyl. In some embodiments, R⁵ is 3-azepanyl. In some embodiments, R⁵ is 4-azepanyl. In some embodiments, R⁵ is 2-morpholinyl. In some embodiments, R⁵ is 2-thiomorpholinyl. In some embodiments, R⁵ is 2-piperazinyl. [00142] In some embodiments, L¹ and L² are each independently

,

—O—, —S(O)_x—, —S—S—, —NR^aC(═O)—, —C(═O)NR^a—, —NR^aC(═O)NR^a—, —OC(═O)NR^a—, or —NR^aC(═O)O—. In some embodiments, L¹ and L² are each independently

_{(C═O)O—. In some embodiments, L1 and L2 are each independently}

_,

, —O(C═O)—, or —(C═O)O—. In some embodiments, L¹ and L² are each independently

or a direct bond. In some embodiments, L¹ and L² are each independently —O(C═O)—, — (C═O)O—, or a direct bond. [00143] In some embodiments, at least one of L¹ and L² (i.e., L¹; L²; L¹ and L²) is

,

some embodiments, L¹ and L² are each independently

some embodiments,

so e e o e s, a o a e . so e e o e s, a o L² are

. , . In some embodiments, L¹ and/or L² are —O(C═O)—. In some embodiments, L¹ and/or L² are —(C═O)O—. In some embodiments, L¹ and/or L² are —C(=O)—. In some embodiments, L¹ and/or L² are —C(=O)—. In some embodiments, L¹ and/or L² are —O—. In some embodiments, L¹ and/or L² are —S(O)_x—. In some embodiments, L¹ and/or L² are —S—S—. In some embodiments, L¹ and/or L² are —NR^aC(═O)—. In some embodiments, L¹ and/or L² are —C(═O)NR^a—. In some embodiments, L¹ and/or L² are —NR^aC(═O)NR^a—. In some embodiments, L¹ and/or L² are —OC(═O)NR^a—. In some embodiments, L¹ and/or L² are —NR^aC(═O)O—. In some embodiments, L¹ and/or L² are a direct bond. [00145] In some embodiments, L¹ and L² are the same. In some embodiments, L¹ and L² are the same, and are each

—(C═O)O—. In some embodiments, L¹ and L² are the same, and are each

; and R¹ and R² are the same, and are each unbranched C1—C10 alkyl. In some embodiments, L¹ and L² are the same, and are each —O(C═O)— or —(C═O)O—; and R¹ and R² are the same, and are each branched C1—C16 alkyl. In some embodiments, L¹ and L² are different. [00146] In some embodiments, L³⁰ is —O(C═O)—, —(C═O)O—, —C(=O)—, or —O—. In some embodiments, L³⁰ is —O(C═O)— or —(C═O)O—. In some embodiments, L³⁰ is —O(C═O)—. In some embodiments, L³⁰ is —C(=O)O—. In some embodiments, L³⁰ is —C(=O)—. In some embodiments, L³⁰ is —O—. In some embodiments, L³⁰ is —S(O)_x3—. In some embodiments, L³⁰ is —S—S—. In some embodiments, L³⁰ is —NR^3aC(═O)—. In some embodiments, L³⁰ is —C(═O)NR^3a—. In some embodiments, L³⁰ is —NR^3aC(═O)NR^3a—. In some embodiments, L³⁰ is —OC(═O)NR^3a—. In some embodiments, L³⁰ is —NR^3aC(═O)O—. [00147] In some embodiments, m is 0 and n is 5. In some embodiments, m is 1 and n is 4. In some embodiments, m is 2 and n is 3. In some embodiments, m is 0 and n is 4. In some embodiments, m is 1 and n is 3. In some embodiments, m is 2 and n is 2. In some embodiments, m is 0 and n is 3. In some embodiments, m is 1 and n is 2. In some embodiments, m is 0 and n is 2. In some embodiments, m is 1 and n is 1. [00148] In some embodiments, G³ is C1—C12 unbranched alkylene. In some embodiments, G³ is C₁—C₁₂ branched alkylene. In some embodiments, G³ is C₁—C₅ alkylene (e.g., C₁—C₅ branched alkylene, C1—C5 unbranched alklyene). [00149] In some embodiments, R^b is C1-C4 aliphatic. In some embodiments, R^b is C1-C4 alkylene. [00150] In some embodiments, R^c is C₁—C₁₂ aliphatic. In some embodiments, R^c is C₁—C₁₂ unbranched aliphatic. In some embodiments, R^c is C1—C12 branched aliphatic. In some embodiments, R^c is C1—C12 alkyl (e.g.¸ C1—C12 branched alkyl, C1—C12 unbranched alkyl). [00151] In some embodiments, G¹ is a direct bond. In some embodiments, G¹ is C₁—C₂ alkylene. In some embodiments, G¹ is C1 alkylene. In some embodiments, G¹ is C2 alkylene. [00152] In some embodiments, M¹ and M² are the same. In some embodiments, M¹ and M² are both a direct bond. In some embodiments, M¹ and M² are both C₁—C₁₂ aliphatic. In some embodiments, M¹ and M² are both C1—C12 branched aliphatic. In some embodiments, M¹ and M² are both C1—C12 unbranched aliphatic. In some embodiments, M¹ and M² are both C6—C12 aliphatic (e.g., C₆—C₁₂ branched aliphatic, C₆—C₁₂ unbranched aliphatic). [00153] In some embodiments, M¹ and M² are each independently C₁—C₁₂ unbranched aliphatic or a direct bond. In some embodiments, M¹ and M² are each independently C1—C12 branched aliphatic or a direct bond. In some embodiments, M¹ and M² are each independently C₁—C₁₂ alkylene (e.g., C₁—C₁₂ branched alkylene, C₁—C₁₂ unbranched alkylene) or a direct bond. [00154] In some embodiments, R¹ and R² are both C1—C16 aliphatic. In some embodiments, R¹ and R² are both C1—C16 branched aliphatic. In some embodiments, R¹ and R² are both C1— C₁₆ unbranched aliphatic. In some embodiments, R¹ and R² are C₁—C₁₆ alkyl (e.g., C₁—C₁₆ branched alkyl, C1—C16 unbranched alkyl). In some embodiments, R¹ and R² are C1—C16 branched alkyl. In some embodiments, R¹ and R² are C1—C10 unbranched alkyl. In some embodiments, R¹ and R² are C₄—C₁₆ alkyl. In some embodiments, R¹ and R² are C₄—C₁₆ branched alkyl. In some embodiments, R¹ and R² are C4—C10 unbranched alkyl. [00155] In some embodiments, R¹ and R² are each independently H or C1—C12 aliphatic. In some embodiments, R¹ and R² are both C₁—C₁₂ aliphatic. In some embodiments, R¹ and R² are both C₁—C₁₂ branched aliphatic. In some embodiments, R¹ and R² are both C₁—C₁₂ unbranched aliphatic. In some embodiments, R¹ and R² are C1—C12 alkyl (e.g., C1—C12 branched alkyl, C1—C12 unbranched alkyl). In some embodiments, R¹ and R² are both H. [00156] In some embodiments, R¹ and R² are the same. In some embodiments, R¹ and R² are the same, and are C1—C16 alkyl (e.g., C1—C16 branched alkyl, C1—C10 unbranched alkyl). In some embodiments, R¹ and R² are different. [00157] In some embodiments, each R^c, R¹, and R² is the same. [00158] In some embodiments, R³ is C4—C20 aliphatic. In some embodiments, R³ is C4—C20 unbranched aliphatic. In some embodiments, R³ is C4—C20 branched aliphatic. In some embodiments, R³ is C₄—C₂₀ alkyl (e.g., C₄—C₂₀ unbranched alkyl, C₄—C₂₀ branched alkyl). In some embodiments, R³ is C₄—C₁₀ aliphatic (e.g., C₄—C₁₀ unbranched aliphatic, C₄—C₁₀ branched aliphatic). In some embodiments, R³ is C10—C20 aliphatic (e.g., C10—C20 unbranched aliphatic, C10—C20 branched aliphatic). In some embodiments, R³ is C4—C10 alkyl (e.g., C4— C₁₀ unbranched alkyl, C₄—C₁₀ branched alkyl). In some embodiments, R³ is

. [00159] In some embodiments, R³⁰ is C1—C12 aliphatic. In some embodiments, R³⁰ is C1—C12 branched aliphatic. In some embodiments, R³⁰ is C1—C12 unbranched aliphatic. In some embodiments, R³⁰ is C₁—C₁₂ alkyl (e.g., C₁—C₈ alkyl). In some embodiments, R³⁰ is H. [00160] In some embodiments,

[00161] In some embodiments, R⁴ is

. In some embodiments, R⁴ is

.

[00162] In some embodiments, G⁴ is C₂—C₁₂ unbranched alkylene. In some embodiments, G⁴ is C2—C12 branched alkylene. In some embodiments, G⁴ is C2—C5 alkylene (e.g., C2—C5 unbranched alkylene, C₂—C₅ branched alkylene). In some embodiments, G⁴ is C₃ alkylene. [00163] In some embodiments, R⁶ and R⁷ are each independently C₁—C₆ aliphatic. In some embodiments, R⁶ and R⁷ are each independently C1—C6 alkyl. In some embodiments, R⁶ and R⁷ are each independently C1—C3 alkyl. In some embodiments, R⁶ and R⁷ are both methyl. [00164] In some embodiments, R⁶ and R⁷ are the same. In some embodiments, R⁶ and R⁷ are different. [00165] In some embodiments, R⁶ and R⁷, taken together with the nitrogen atom to which they are attached, form a 4-, 5-, 6-, or 7-membered, nitrogen-containing heterocycle, wherein each nitrogen atom of the heterocycle is independently substituted with R^d, and the heterocycle is optionally substituted on a carbon atom with R^e. In some embodiments, R⁶ and R⁷, taken together with the nitrogen atom to which they are attached, form azetidinyl, pyrrolidinyl, piperdinyl, azepanyl, morpholinyl, thiomorpholinyl, or piperazinyl, wherein each nitrogen atom of the heterocycle is independently substituted with R^d, and the heterocycle is optionally substituted on a carbon atom with R^e. [00166] In some embodiments, G⁴ is C₂—C₅ alkylene; and R⁶ and R⁷ are each C₁—C₃ alkyl and are the same; or R⁶ and R⁷ taken together with the nitrogen atom to which they are attached, form a 4-, 5-, 6-, or 7-membered, nitrogen-containing heterocycle, wherein each nitrogen atom of the heterocycle is independently substituted with R^d, and the heterocycle is optionally substituted on a carbon atom with R^e. [00167] In some embodiments, each nitrogen atom of the heterocycle formed by R⁶ and R⁷ taken together with the nitrogen atom to which they are attached other than the nitrogen atom attached to G⁴, R⁶, and R⁷, is independently substituted with R^d. In some embodiments, one or more nitrogen atoms (e.g., one nitrogen atom) of the heterocycle formed by R⁶ and R⁷ taken together with the nitrogen atom to which they are attached other than the nitrogen atom attached to G⁴, R⁶, and R⁷, are independently substituted with R^d. [00168] In some embodiments, R^a is H. In some embodiments, R^a is C1—C12 aliphatic. In some embodiments, R^a is C1—C12 alkyl. In some embodiments, R^a is C6—C12 aliphatic. In some embodiments, R^a is C₆—C₁₂ alkyl. [00169] In some embodiments R^b is C1—C8 alkylene. In some embodiments R^b is C1—C4 aliphatic. In some embodiments R^b is C1—C4 alkylene. In some embodiments R^b is C2 aliphatic. In some embodiments R^b is C₁ aliphatic. [00170] In some embodiments, R^c is H. In some embodiments, R^c is C1—C12 aliphatic. In some embodiments, R^c is C1—C12 alkyl. In some embodiments, R^c is C6—C12 aliphatic. In some embodiments, R^c is C₆—C₁₂ alkyl. [00171] In some embodiments R^d is C₁—C₆ alkyl or (C₁—C₆ alkyl)OH. In some embodiments R^d is C1—C3 alkyl or (C1—C3 alkyl)OH. In some embodiments, R^d is methyl or hydroxyethyl. In some embodiments R^d is C1—C6 aliphatic. In some embodiments R^d is C1—C6 alkyl. In some embodiments R^d is C₁—C₃ aliphatic. In some embodiments R^d is C₁—C₃ alkyl. In some embodiments R^d is methyl. In some embodiments R^d is ethyl. In some embodiments, R^d is (C1— C6 aliphatic)OH. In some embodiments, R^d is (C1—C6 alkyl)OH. In some embodiments, R^d is (C₁—C₃ aliphatic)OH. In some embodiments, R^d is (C₁—C₃ alkyl)OH. In some embodiments, R^d is hydroxyethyl. In some embodiments, R^d is hydroxymethyl. [00172] In some embodiments, R^e is -OH. In some embodiments, R^e is (C1—C6 aliphatic)OH. In some embodiments, R^e is (C₁—C₆ alkyl)OH. In some embodiments, R^e is (C₁—C₃ aliphatic)OH. In some embodiments, R^e is (C₁—C₃ alkyl)OH. In some embodiments, R^e is hydroxyethyl. In some embodiments, R^e is hydroxymethyl. In some embodiments, R^e is -O(C1— C6 aliphatic). In some embodiments, R^e is -O(C1—C6 alkyl). In some embodiments, R^e is - O(C₁—C₃ aliphatic). In some embodiments, R^e is -O(C₁—C₃ alkyl). In some embodiments, R^e is ethoxy. In some embodiments, R^e is methoxy. [00173] In some embodiments R^3a is H. In some embodiments R^3a is C1—C12 aliphatic. In some embodiments R^3a is C₁—C₁₂ alkyl. In some embodiments R^3a is C₁—C₆ aliphatic. In some embodiments, R^3a is C1—C6 alkyl (e.g., methyl, ethyl, or propyl). [00174] In some embodiments, x is 0. In some embodiments, x is 1. In some embodiments, x is 2. [00175] In some embodiments, x3 is 0. In some embodiments, x3 is 1. In some embodiments, x3 is 2. [00176] In some embodiments, is

[00177] Another embodiment provides a compound of Table 1, or a pharmaceutically acceptable salt, solvate, or hydrate thereof.

[00178] Methods of making specific compounds of formula (I) are described herein in the Examples. Scheme 1 sets forth a general procedure for making compounds of formula (I), wherein the variables are as described for a compound of formula (I) herein. Scheme 1

[00179] As set forth generally in Scheme 1, sequential alkylation of the compound of structure A with A’ and A” (e.g., alkylation with A’ followed by alkylation with A”; alkylation with A” followed by alkylation with A’) forms a compound of structure B, which, upon hydrolysis (e.g., under acidic conditions, such as in the presence of HCl), leads to a compound of structure C. Alkylation of the compound of structure A can be conducted under basic conditions, e.g., using sodium hydride and tetra-N-butylammonium iodide (TBAI). M¹ and M² in the compound of structure C are connected to R¹ and R² via linkers L¹ and L² (e.g., using various transformations, such as esterification, amide formation) to form a compound of structure D. The ketone in structure D can undergo an extension to form a compound of structure E (e.g. via Wittig reaction followed by reduction (for 2-atom extension) or Darzens reaction (for 1-atom extension)). Reductive amination of a compound of structure E, produces a compound of structure F, which can be reacted with an appropriate electrophile, for example, a carbamoyl chloride, alkyl sulfonyl chloride, or alkyl sulfinyl chloride, (e.g., in the presence of (tri)phosgene) to produce a compound of formula (I). Lipid Particles [00180] Compounds described herein can be in the form of lipid particles, such as lipid nanoparticles (LNPs), micelles and liposomes. Lipid particles, such as lipid nanoparticles, can be unilamellar or multilamellar. [00181] Thus, one embodiment is a lipid particle (e.g., a lipid nanoparticle, a liposome) comprising one or more compounds of the disclosure. In some embodiments, a lipid particle (e.g., LNP) further comprises a therapeutic agent. In some embodiments, a lipid particle (e.g., LNP) further comprises one or more nucleic acids. In some embodiments, a lipid particle (e.g., LNP) comprises one or more of: a steroid, a polymer-conjugated lipid (e.g., PEG-lipid), permanent cationic lipid, and a phospholipid. [00182] Another embodiment provides a lipid particle (e.g.¸ LNP) comprising a compound of the disclosure, steroid, polymer-conjugated lipid (e.g., PEG-lipid), and phospholipid. In some embodiments, the lipid particle further comprises a therapeutic agent and/or one or more nucleic acids. [00183] Yet another embodiment provides a lipid particle (e.g., LNP) comprising: (a) one or more therapeutic agents or nucleic acids; (b) one or more ionizable cationic lipids of the disclosure, wherein the one or more ionizable cationic lipids of the disclosure are from about 30 mol % to about 85 mol % of total lipid present in the particle; (c) one or more steroids, wherein the one or more steroids are from about 30 mol % to about 45 mol % of the total lipid present in the particle; (d) one or more phospholipids, wherein the one or more phospholipids are from about 6 mol % to about 35 mol % of the total lipid present in the particle; and (e) one or more PEG lipids, wherein the one or more PEG lipids are from about 0.5 mol % to about 3 mol % of the total lipid present in the particle. In some embodiments, the one or more ionizable cationic lipids are about 50 mol % of the total lipids present in the particle, the one or more steroids are about 38.5 mol % of the total lipids present in the particle, the one or more phospholipids are about 10 mol % of the total lipids present in the particle, and the one or more PEG lipids are about 1.5% of the total lipids present in the particle. [00184] The lipid particles (e.g., LNPs) described herein typically have a mean diameter of from about 40 nm to about 150 nm. In some embodiments, the lipid particles have a mean diameter of from about 40 nm to about 100 nm. In some embodiments, the lipid particles have a mean diameter of from about 40 nm to about 80 nm. In some embodiments, the lipid particles have a mean diameter of from about 40 nm to about 70 nm. In some embodiments, the lipid particles have a mean diameter of from about 50 nm to about 70 nm. In some embodiments, the lipid particles have a mean diameter of from about 60 nm to about 70 nm. In some embodiments, the lipid particles have a mean diameter of from about 50 nm to about 60 nm. In some embodiments, the lipid particles have a mean diameter of from about 50 nm to about 150 nm. In some embodiments, the lipid particles have a mean diameter of from about 60 nm to about 130 nm. In some embodiments, the lipid particles have a mean diameter of from about 70 nm to about 110 nm. Particle size can be measured using dynamic light scattering, for example, in accordance with the Examples. [00185] N:P ratio can be used to described the molar ratio of basic nitrogens (N) (e.g., from ionizable cationic lipids) to acidic phosphates (P) (e.g., from the nucleic acid backbone) in lipid particles. The N:P ratio is commonly used as a proxy for the average ratio of positive ions (nitrogens, N) to negative ions (phosphates, P) in a lipid particle such as those described herein. High N:P particles or formulations have more lipid relative to nucleic acid than low N:P formulations. In some embodiments, the N:P ratio of a lipid particle is from about 2 to about 10 or about 3 to about 7. In some embodiments, the N:P ratio of a lipid particle is about 6. Ionizable cationic lipids [00186] Ionizable cationic lipids are often the major lipid component in lipid nanoparticles (LNPs). They are designed to be charge-neutral at standard physiological pH (~7) but acquire a positive charge at lower (more acidic) pH. This helps with nucleic acid payload encapsulation during LNP formation and intracellular delivery (the endosomal fusion step is believed to require a positive charge on the ionizable cationic lipid). [00187] The lipid particles of the present disclosure comprise a compound of the disclosure. In some embodiments, a lipid particle further comprises an additional ionizable cationic lipid. The additional ionizable cationic lipid may be an additional compound of the disclosure or it may be a lipid that is not described by formula (I). Suitable additional ionizable lipids for use in accordance with this technology include 1,2-dilinoleyoxy-3-(dimethylamino)acetoxypropane (DLin-DAC), 1,2-dilinoleyoxy-3morpholinopropane (DLin-MA), 1,2-dilinoleoyl-3- dimethylaminopropane (DLinDAP), 1,2-dilinoleylthio-3-dimethylaminopropane (DLin-S- DMA), 1-linoleoyl-2-linoleyloxy-3dimethylaminopropane (DLin-2-DMAP), 1,2-dilinoleyloxy- 3-trimethylaminopropane chloride salt (DLin-TMA Cl), 1,2-dilinoleoyl-3- trimethylaminopropane chloride salt (DLin-TAP Cl), 1,2-dilinoleyloxy-3-(N- methylpiperazino)propane (DLin-MPZ), 3-(N,Ndilinoleylamino)-1,2-propanediol (DLinAP), 3- (N,N-dioleylamino)-1,2-propanediol (DOAP), 1,2-dilinoleyloxo-3-(2-N,N- dimethylamino)ethoxypropane (DLin-EG-DMA), and 2,2-dilinoleyl-4-dimethylaminomethyl- [1,3]-dioxolane (DLin-K-DMA). [00188] In some embodiments, an ionizable cationic lipid (e.g. of formula (I) or a subformula thereof) is from about 1% to about 90% of the total lipids in a lipid particle of the present disclosure on a molar basis. In some embodiments, an ionizable cationic lipid (e.g. of formula (I) or a subformula thereof) is from about 5% to about 90%, from about 10% to about 90%, from about 15% to about 90%, from about 20% to about 90%, from about 25% to about 90%, from about 30% to about 90%, from about 35% to about 90%, from about 40% to about 90%, from about 45% to about 90%, from about 50% to about 90%, from about 55% to about 90%, from about 60% to about 90%, from about 65% to about 90%, from about 70% to about 90%, from about 75% to about 90%, from about 80% to about 90%, or from about 85% to about 90%, of the total lipids in a lipid particle of the present disclosure on a molar basis. [00189] In some embodiments, an ionizable cationic lipid (e.g. of formula (I) or a subformula thereof) is from about 1% to about 85% of the total lipids in a lipid particle of the present disclosure on a molar basis. In some embodiments, an ionizable cationic lipid (e.g. of formula (I) or a subformula thereof) is from about 1% to about 80%, from about 1% to about 75%, from about 1% to about 70%, from about 1% to about 65%, from about 1% to about 60%, from about 1% to about 55%, from about 1% to about 50%, from about 1% to about 45%, from about 1% to about 40%, from about 1% to about 35%, from about 1% to about 30%, from about 1% to about 25%, from about 1% to about 20%, from about 1% to about 15%, from about 1% to about 10%, or from about 1% to about 5%, of the total lipids in a lipid particle of the present disclosure on a molar basis. [00190] In some embodiments, an ionizable cationic lipid (e.g. of formula (I) or a subformula thereof) is from about 5% to about 85% of the total lipids in a lipid particle of the present disclosure on a molar basis. In some embodiments, an ionizable cationic lipid (e.g. of formula (I) or a subformula thereof) is from about 10% to about 80%, from about 15% to about 75%, from about 20% to about 70%, from about 25% to about 65%, from about 30% to about 60%, from about 30% to about 65%, from about 30% to about 70%, from about 35% to about 65%, from about 40% to about 60%, from about 45% to about 55%, or about 50%, of the total lipids in a lipid particle of the present disclosure on a molar basis. In some embodiments, an ionizable cationic lipid (e.g., of Formula (I) or a subformula thereof) is from about 20% to about 60% of total lipids in the lipid particle of the present disclosure on a molar basis. In some embodiments, an ionizable cationic lipid (e.g. of formula (I) or a subformula thereof) is about 50% of the total lipids in a lipid particle of the present disclosure on a molar basis. Permanent cationic lipids [00191] In some embodiments, a lipid particle comprises a permanent cationic lipid. The term “permanent cationic lipid,” as used herein, refers to a lipid that contains a positive charge insensitive to environmental pH. Permanent cationic lipids typically do not possess a positive charge because of a protonation, which can be deprotonated at a higher pH, and instead are often positively charged due to the alkylation of a lone pair on an otherwise neutral atom, such as nitrogen or sulfur. [00192] Examples of suitable permanent cationic lipids for use in accordance with the present disclosure include, but are not limited to: N-(2,3-dioleyloxy)propyl)-N,N,N-trimethylammonium chloride (DOTMA); N,N-distearyl-N,N-dimethylammonium bromide (DDAB); N- (2,3dioleoyloxy)propyl)-N,N,N-trimethylammonium chloride (DOTAP); N-(1-(2,3- dioleoyloxy)propyl)N-2-(sperminecarboxamido)ethyl)-N,N-dimethylammonium trifluoracetate (DOSPA); O,O’-ditetradecanoyl-N-(α-trimethylammonioacetyl)diethanolamine chloride (DC-6- 14); N-(2-hydroxyethyl)-N,N-dimethyl-2,3-bis(oleoyloxy)propan-1-aminium bromide (DORI); N-(1,2dimyristyloxyprop-3-yl)-N,N-dimethyl-N-hydroxyethyl ammonium bromide (DMRIE); and O-alkyl phosphatidylcholine derivatives such as 1,2-dioleoyl-sn-glycero-3- ethylphosphocholine chloride (18:1 EPC). [00193] In some embodiments, permanent cationic lipids are less than about 5% of the cationic lipids in a lipid particle of the present disclosure on a molar basis. In some embodiments, permanent cationic lipids are less than about 10%, less than about 20%, less than about 30%, less than about 40%, less than about 50%, less than about 60%, less than about 70%, less than about 80%, or less than about 90%, of the cationic lipids in a lipid particle of the present disclosure on a molar basis. Phospholipids [00194] In some embodiments, a lipid particle further comprises a phospholipid. Phospholipids suitable for use in accordance with the present disclosure include, but are not limited to, phosphatidylserine (PS), phosphatidylcholine (PC), phosphatidylethanolamine (PE), and phosphatidylinositol (PI), among others. Additional examples of phospholipids include dimyristoylphosphatidylcholine (DMPC), 1,2-dioleoyl-sn-glycero-3-phosphocholine 18:1 Δ9- Cis PC (DOPC), 1,2-distearoyl-sn-glycero-3-phosphocholine 18:0 (DSPC), 1-palmitoyl-2- oleoyl-glycero-3-phosphocholine 16:0-18:1 (POPC), bisphosphatidyl glycerol, phosphatidic acid, phosphatidyl alcohol, phosphatidyl glycerol, lecithin, lysolecithin, lysophosphatidylethanolamine, sphingomyelin, egg sphingomyelin (ESM), cephalin, cardiolipin, phosphatidic acid, cerebrosides, dicetylphosphate, dipalmitoylphosphatidylcholine (DPPC), dioleoylphosphatidylglycerol (DOPG), dipalmitoylphosphatidylglycerol (DPPG), dioleoylphosphatidylethanolamine (DOPE), palmitoyloleoyl-phosphatidylethanolamine (POPE), palmitoyloleyol-phosphatidylglycerol (POPG), dioleoylphosphatidylethanolamine 4-(N- maleimidomethyl)-cyclohexane-1-carboxylate (DOPE-mal), dipalmitoyl- phosphatidylethanolamine (DPPE), dimyristoyl- phosphatidylethanolamine (DMPE), distearoyl- phosphatidylethanolamine (DSPE), monomethyl-phosphatidylethanolamine, dimethyl- phosphatidylethanolamine, dielaidoyl- phosphatidylethanolamine (DEPE), stearoyloleoyl- phosphatidylethanolamine (SOPE), lysophosphatidylcholine, dilinoleoylphosphatidylcholine, and mixtures thereof. In some embodiments, a phospholipid is dioleoylphosphatidyl ethanolamine (DOPE), dimyristoylphosphatidyl choline (DMPC), distearoylphosphatidyl choline (DSPC), dimyristoylphosphatidyl glycerol (DMPG), dipalmitoyl phosphatidylcholine (DPPC), or phosphatidylcholine (PC). [00195] Phospholipids can be saturated or unsaturated, i.

., contain one or more units of unsaturation, and can contain acyl chains of a variety of lengths. In some aspects, a phospholipid contains a C4-C30 acyl chain, e.g., a C8-C26, C12-C22, C10-C25, C14-C18 or C16-C26 acyl chain. Phospholipids can be obtained from various sources, both natural and synthetic. For example, PS can be obtained from porcine brain PS or plant-based soy (soya bean) PS. Egg PC and PS and synthetic PC are available commercially. [00196] In some embodiments, a phospholipid is from about 1% to about 90% of the total lipids in a lipid particle of the present disclosure on a molar basis. In some embodiments, a phospholipid is from about 5% to about 90%, from about 10% to about 90%, from about 15% to about 90%, from about 20% to about 90%, from about 25% to about 90%, from about 30% to about 90%, from about 35% to about 90%, from about 40% to about 90%, from about 45% to about 90%, from about 50% to about 90%, from about 55% to about 90%, from about 60% to about 90%, from about 65% to about 90%, from about 70% to about 90%, from about 75% to about 90%, from about 80% to about 90%, or from about 85% to about 90%, of the total lipids in a lipid particle of the present disclosure on a molar basis. [00197] In some embodiments, a phospholipid is from about 1% to about 85% of the total lipids in a lipid particle of the present disclosure on a molar basis. In some embodiments, a phospholipid is from about 1% to about 80%, from about 1% to about 75%, from about 1% to about 70%, from about 1% to about 65%, from about 1% to about 60%, from about 1% to about 55%, from about 1% to about 50%, from about 1% to about 45%, from about 1% to about 40%, from about 1% to about 35%, from about 1% to about 30%, from about 1% to about 25%, from about 1% to about 20%, from about 1% to about 15%, from about 1% to about 10%, or from about 1% to about 5%, of the total lipids in a lipid particle of the present disclosure on a molar basis. [00198] In some embodiments, a phospholipid is from about 0.1% to about 40% of the total lipids in a lipid particle of the present disclosure on a molar basis. In some embodiments, a phospholipid is from about 1% to about 35%, from about 2.5% to about 30%, from about 5% to about 25%, from about 7.5% to about 20%, from about 8% to about 15%, from about 8% to about 12.5%, or about 10%, of the total lipids in a lipid particle of the present disclosure on a molar basis. In some embodiments, phospholipids are about 10% of the total lipids in a lipid particle of the present disclosure on a molar basis. [00199] In some embodiments, a phospholipid (taken together or each individually) is from about 6 percent to about 35 percent of the total lipids in the lipid particle on a molar basis. In some embodiments, a phospholipid (taken together or each individually) is about 10 percent of the total lipids in the lipid particle on a molar basis. Steroid [00200] In some embodiments, a lipid particle (e.g., LNP) further comprises a steroid. Steroids suitable for use in accordance with the present disclosure include, but are not limited to, cholesterol and derivatives thereof, such as cholestanol, cholestanone, cholestenone, coprostanol, cholesteryl-2′- hydroxyethyl ether, cholesteryl-4′-hydroxybutyl ether, and mixtures thereof. Other steroids suitable for inclusion in the lipid particles described herein include N⁴- cholesteryl-spermine, or a salt thereof, such as N⁴-cholesteryl-spermine HCl salt. N⁴- cholesteryl-spermine HCl salt is also known as Genzyme Lipid 67 and is a cholesterol derivatized with spermine to create a cationic lipid HCl salt. In some embodiments, a steroid is cholesterol. [00201] In some embodiments, a steroid is from about 1% to about 90% of the total lipids in a lipid particle of the present disclosure on a molar basis. In some embodiments, a steroid is from about 5% to about 90%, from about 10% to about 90%, from about 15% to about 90%, from about 20% to about 90%, from about 25% to about 90%, from about 30% to about 90%, from about 35% to about 90%, from about 40% to about 90%, from about 45% to about 90%, from about 50% to about 90%, from about 55% to about 90%, from about 60% to about 90%, from about 65% to about 90%, from about 70% to about 90%, from about 75% to about 90%, from about 80% to about 90%, or from about 85% to about 90%, of the total lipids in a lipid particle of the present disclosure on a molar basis. [00202] In some embodiments, a steroid is from about 1% to about 85% of the total lipids in a lipid particle of the present disclosure on a molar basis. In some embodiments, a steroid is from about 1% to about 80%, from about 1% to about 75%, from about 1% to about 70%, from about 1% to about 65%, from about 1% to about 60%, from about 1% to about 55%, from about 1% to about 50%, from about 1% to about 45%, from about 1% to about 40%, from about 1% to about 35%, from about 1% to about 30%, from about 1% to about 25%, from about 1% to about 20%, from about 1% to about 15%, from about 1% to about 10%, or from about 1% to about 5%, of the total lipids in a lipid particle of the present disclosure on a molar basis. [00203] In some embodiments, a steroid is from about 1% to about 80% of the total lipids in a lipid particle of the present disclosure on a molar basis. In some embodiments, a steroid is from about 5% to about 75%, from about 10% to about 70%, from about 15% to about 65%, from about 20% to about 60%, from about 25% to about 55%, from about 30% to about 50%, from about 35% to about 45%, about 40%, or about 38.5% of the total lipids in a lipid particle of the present disclosure on a molar basis. In some embodiments, a steroid is from about 30% to about 45% of the total lipids in a lipid particle on a molar basis. In some embodiments, steroids are about 38.5% of the total lipids in a lipid particle of the present disclosure on a molar basis. Polymer-Conjugated Lipids [00204] In some embodiments, a lipid particle (e.g., LNP) further comprises a polymer- conjugated lipid (e.g., PEG-lipid). Suitable polymer-conjugated lipids for use in accordance with the present disclosure include, but are not limited to, polyethyleneglycol (PEG)-lipids, polyamide (ATTA)-lipid conjugates, cationic-polymer-lipid conjugates (CPLs), polyoxazoline (POZ)-lipids, and polysarcosine (pSAR)-lipids. [00205] In some embodiments, the lipid nanoparticles comprise a PEG-lipid or an ATTA-lipid conjugate. In some embodiments, the lipid nanoparticles comprise a PEG-lipid. A PEG-lipid may comprise a PEG-diacylglycerol (DAG), a PEG succinate diacylglycerol (PEG-S-DAG) such as 4-O-(2′,3′-di(tetradecanoyloxy)propyl-1-O-(ω-methoxy(polyethoxy)ethyl)butanedioate (PEG-S-DMG), a PEG dialkyloxypropyl (DAA), a PEG-phospholipid (e.g., PEG- phosphatidylethanoloamine (PEG-PE)), a PEG-ceramide (Cer), or mixtures thereof. The PEG- DAA conjugate may be PEG-dilauryloxypropyl (C12), PEG-dimyristyloxypropyl (C14), PEG- dipalmityloxypropyl (C16), PEG-distearyloxypropyl (C18), or mixtures thereof. Additional PEG- lipids suitable for use in accordance with the present disclosure include, but are not limited to, mPEG2000-1,2-di-O-alkyl-sn3-carbomoylglyceride (PEG-C-DOMG), 1- [8′-(1,2-dimyristoyl-3- propanoxy)-carboxamido-3′,6′-dioxaoctanyl]carbamoyl-ω-methyl-poly(ethylene glycol) (DMG, DMG-PEG, DMG-PEG 2000). [00206] PEG is a linear, water-soluble polymer of ethylene glycol repeating units. PEGs are classified by their molecular weights; for example, PEG 2000 has an average molecular weight of about 2,000 daltons, and PEG 5000 has an average molecular weight of about 5,000 daltons. PEG compounds suitable for conjugation to lipids are commercially available from multiple chemical suppliers and include, for example, the following: monomethoxypolyethylene glycol (MePEG-OH), monomethoxypolyethylene glycol-succinate (MePEG-S), monomethoxypolyethylene glycol-succinimidyl succinate (MePEG-S-NHS), monomethoxypolyethylene glycol-amine (MePEG- NH2), monomethoxypolyethylene glycol- tresylate (MePEG-TRES), and monomethoxypolyethylene glycol-imidazolyl-carbonyl (MePEG- IM). [00207] In some embodiments, the PEG of a PEG lipid has an average molecular weight of from about 550 daltons to about 10,000 daltons. In certain embodiments, the PEG of a PEG lipid has an average molecular weight of from about 750 daltons to about 5,000 daltons (e.g., from about 1,000 daltons to about 5,000 daltons, from about 1,500 daltons to about 3,000 daltons, from about 750 daltons to about 3,000 daltons, from about 750 daltons to about 2,000 daltons, etc.). In preferred embodiments, the PEG moiety has an average molecular weight of about 2,000 daltons or about 750 daltons. [00208] In certain instances, the polymer-conjugated lipid (e.g., PEG lipid) is from about 0.1 to about 10% (or any fraction thereof or range therein) of the total lipid present in the particle on a molar basis. In certain instances, the polymer-conjugated lipid (e.g., PEG lipid) is from about 0.1% to about 5% (or any fraction thereof or range therein) of the total lipid present in the particle on a molar basis. In certain instances, the polymer-conjugated lipid (e.g., PEG lipid) is from about 0.1% to about 2%, from about 0.5% to about 2%, from about 1% to about 2l%, from about 0.6% to about 1.9%, from about 0.7% to about 1.8%, from about 0.8% to about 1.7%, from about 1% to about 1.8%, from about 1.2% to about 1.8%, from about 1.2% to about 1.7%, from about 1.3% to about 1.6%, from about 1.4% to about 1.5%, or about: 1, 1.1, 1.2, 1.3, 1.4, 1.5, 1.6, 1.7, 1.8, 1.9, or 2% (or any fraction thereof or range therein) of the total lipid present in the particle on a molar basis. In some embodiments, a polymer-conjugated lipid (e.g., PEG lipid) is from about 0.5 percent to about 3 percent of total lipids present in the particle on a molar basis. In some embodiments, a polymer-conjugated lipid (e.g., PEG lipid) is about 1.5 percent of total lipids present in the particle on a molar basis. Therapeutic agents and nucleic acids [00209] In some embodiments, a lipid particle further comprises one or more nucleic acids. [00210] As used herein, the term “nucleic acid” includes oligonucleotides and polynucleotides. Non-limiting examples of nucleic acids suitable for use in accordance with the present disclosure include interfering RNA molecules (e.g., siRNA, aiRNA, miRNA, antisense RNA, tRNA), mRNA, DNA, plasmids, and immunostimulatory oligonucleotides. Additional nucleic acids suitable for use in accordance with the present disclosure include siRNA, dsDNA, mRNA, and plasmids, including plasmids from which an interfering RNA or mRNA can be transcribed. [00211] In some embodiments, the one or more nucleic acids comprises DNA. In some embodiments, the one or more nucleic acids comprises RNA (e.g., mRNA, siRNA). In some embodiments, the one or more nucleic acids comprise mRNA, siRNA, or DNA. In some embodiments, the one or more nucleic acids comprise a guide RNA and mRNA (e.g., a cas9 mRNA). [00212] Typically, nucleic acids containing up to 60 nucleotides are referred to as oligonucleotides, and longer nucleic acids are referred to as polynucleotides. In some embodiments, the nucleic acid(s) is an oligonucleotide. In some embodiments, the nucleic acid(s) is a polynucleotide. In some embodiments, an oligonucleotide is from about 15 to about 60 nucleotides in length. [00213] Used herein, the terms “polynucleotide” and “oligonucleotide” include a polymer or oligomer of nucleotide or nucleoside monomers consisting of naturally occurring bases, sugars and intersugar (backbone) linkages, as well as polymers or oligomers comprising non-naturally occurring monomers, or portions thereof, which function similarly. Modified or substituted oligonucleotides may be preferred over native forms because of properties such as, for example, enhanced cellular uptake, reduced immunogenicity, and increased stability in the presence of nucleases. [00214] Nucleic acids of the invention may be of various lengths, generally dependent upon the form of nucleic acid. For example, in some embodiments, the nucleic acid may be from about 60 to about 30,000, from about 60 to about 15,000, from about 60 to about 5,000, from about 60 to about 2000, or from about 60 to about 1000 nucleotide residues in length. [00215] In some embodiments, plasmids or genes may be from about 100 to about 50,000, from about 200 to about 50,000, from about 500 to about 50,000, from about 1000 to about 50,000, from about 5,000 to about 50,000, from about 10,000 to about 50,000, or from about 20,000 to about 50,000 nucleotide residues in length. [00216] In some embodiments, oligonucleotides may range from about 10 to about 60 nucleotides in length. In various related embodiments, oligonucleotides, both single-stranded, double-stranded, and triple-stranded, may range in length from about 10 to about 60 nucleotides, from about 15 to about 60 nucleotides, from about 20 to about 50 nucleotides, from about 15 to about 30 nucleotides, or from about 20 to about 30 nucleotides in length. [00217] In some embodiments, the nucleic acid encodes an engineered nuclease wherein the nuclease comprises a DNA binding domain and a cleavage domain. In some embodiments, the DNA binding domain is a zinc finger DNA binding domain, a TALE DNA binding domain, an RNA molecule (e.g., single guide (sg) RNA of a CRISPR/Cas nuclease), or a meganuclease DNA binding domain. [00218] In some embodiments, the nucleic acid encodes an antigen. In some embodiments, the nucleic acid encodes one or more antigens. In some embodiments, the composition further comprises an adjuvant. In some embodiments, the nucleic acid encodes at least one adjuvant. [00219] In some embodiments, a lipid particle further comprises a therapeutic agent. Non- limiting examples of therapeutic agents include nucleic acids, peptides, polypeptides, and small molecules. Examples of peptides or polypeptides include, without limitation, antibodies, cytokines, growth factors, apoptotic factors, nucleases, differentiation-inducing factors, cell- surface receptors and their ligands, and hormones. Examples of small molecules include, but are not limited to, small organic molecules, particularly those which are hydrophobic. [00220] In some embodiments, the therapeutic agent is a nucleic acid, such as any of the nucleic acids described herein. [00221] In some embodiments, the therapeutic agent is a peptide or polypeptide. In some embodiments, the peptide or polypeptide is an antibody such as, e.g., a polyclonal antibody, a monoclonal antibody, an antibody fragment; a humanized antibody, a recombinant antibody, a recombinant human antibody, a Primatized™ antibody, or mixtures thereof. In some embodiments, the peptide or polypeptide is a cytokine, a growth factor, an apoptotic factor, a differentiation-inducing factor, a cell-surface receptor, a ligand, a nuclease (e.g. a CRISPR/Cas system), or a hormone. [00222] In some embodiments, the therapeutic agent is a small molecule. [00223] In some embodiments, the therapeutic agent or nucleic acid is encapsulated within the lipid nanoparticle. It is believed that encapsulation of the therapeutic agent or nucleic acid in the lipid nanoparticle renders the therapeutic agent or nucleic acid resistant to enzymatic degradation, e.g., by a nuclease or protease, in aqueous solution. In some embodiments, the therapeutic agent or nucleic acid is not substantially degraded after exposure of the particle to a nuclease at 37° C for at least about 20, about 30, about 45, or about 60 minutes. In some embodiments, the therapeutic agent or nucleic acid is not substantially degraded after incubation of the particle in serum at 37° C for at least about: 30, 45, or 60 minutes, or 2, 3, 4, 5, 6, 7, 8, 9, 10, 12, 14, 16, 18, 20, 22, 24, 26, 28, 30, 32, 34, or 36 hours. In other embodiments, the therapeutic agent or nucleic acid (e.g., mRNA or siRNA) is complexed with the lipid particle. [00224] Methods for preparing lipid particles are known to those of skill in the art and described herein in the Examples. [00225] Provided herein in an embodiment is a method for preparing a lipid particle, such as any of the lipid particles described herein. The method comprises providing a mixture of the ionizable cationic lipid(s) and optionally the steroid(s), phospholipid(s), polymer-conjugated lipid(s), and additional cationic lipid(s), in a first solvent; providing a solution of therapeutic agent or nucleic acid in a second solvent; and mixing the mixture and the solution, thereby preparing the lipid particle. In some embodiments, the method further comprises removing the first solvent, for example, by dialysis. In some embodiments, the mixture and the solution are mixed using a microfluidic device. In some embodiments, the first solvent is an alcohol, such as ethanol. In some embodiments, the second solvent is an aqueous buffer, such as an acidic aqueous buffer, e.g., acetate buffer. Compositions [00226] Typically, for administration to a subject, a compound of the disclosure, e.g., in the form of a lipid particle, such as an LNP, is formulated with one or more pharmaceutically acceptable carriers. The disclosure provides such compositions, including pharmaceutical compositions. Thus, one embodiment is a composition (e.g., pharmaceutical composition) comprising a compound of the disclosure, e.g., in the form of a lipid particle, and a pharmaceutically acceptable carrier. One embodiment is a composition (e.g., pharmaceutical composition) comprising a plurality of lipid particles and a pharmaceutically acceptable carrier. The compositions described herein can be used in the methods described herein, e.g., to administer a therapeutic agent, for example, a nucleic acid. [00227] Compositions described herein and, hence, compounds of the disclosure, may be administered orally, parenterally (including subcutaneously, intramuscularly, intravenously and intradermally), by inhalation spray, nasally, or via an implanted reservoir. The terms “parenteral” and “parenterally,” as used herein, include subcutaneous, intracutaneous, intravenous, intramuscular, intraocular, intravitreal, intra-articular, intra-arterial, intra-synovial, intrasternal, intrathecal, intralesional, intrahepatic, intraperitoneal, intralesional and intracranial injection or infusion techniques. In some aspects, a composition described herein is administrable intravenously and/or intraperitoneally. In some aspects, a composition described herein is administrable orally. In some aspects, a composition described herein is administrable subcutaneously. In some aspects, a composition described herein is administrable by inhalation. Preferably, a composition described herein is administered subcutaneously, intraperitoneally, by inhalation or intravenously. Administration can be local or systemic. The preferred mode of administration can vary depending on the particular composition or agent. [00228] Compositions provided herein can be orally administered in any orally acceptable dosage form including, but not limited to, capsules, tablets, aqueous suspensions, dispersions and solutions. In the case of tablets for oral use, carriers commonly used include lactose and corn starch. Lubricating agents, such as magnesium stearate, are also typically added. For oral administration in a capsule form, useful diluents include lactose and dried cornstarch. When aqueous suspensions and/or emulsions are required for oral use, the active ingredient can be suspended or dissolved in an oily phase and combined with emulsifying and/or suspending agents. If desired, certain sweetening, flavoring or coloring agents may also be added. [00229] In some embodiments, an oral formulation is formulated for immediate release or sustained/delayed release. [00230] Solid dosage forms for oral administration include capsules, tablets, pills, powders, and granules. In such solid dosage forms, the active compound is mixed with at least one inert, pharmaceutically acceptable excipient or carrier such as sodium citrate or dicalcium phosphate and/or (a) fillers or extenders such as starches, lactose, sucrose, glucose, mannitol, and silicic acid, (b) binders, such as carboxymethylcellulose, alginates, gelatin, polyvinylpyrrolidinone, sucrose, and acacia, (c) humectants such as glycerol, (d) disintegrating agents such as agar-agar, calcium carbonate, potato or tapioca starch, alginic acid, certain silicates, and sodium carbonate, (e) solution retarding agents such as paraffin, (f) absorption accelerators such as quaternary ammonium salts, (g) wetting agents, such as acetyl alcohol and glycerol monostearate, (h) absorbents such as kaolin and bentonite clay, and (i) lubricants such as talc, calcium stearate, magnesium stearate, solid polyethylene glycols, sodium lauryl sulfate, and mixtures thereof. In the case of capsules, tablets and pills, the dosage form may also comprise buffering agents. [00231] Liquid dosage forms for oral administration include pharmaceutically acceptable emulsions, microemulsions, solutions, suspensions, syrups, and elixirs. Liquid dosage forms may contain inert diluents commonly used in the art, such as water or other solvents, solubilizing agents and emulsifiers, such as ethyl alcohol (ethanol), isopropyl alcohol, ethyl carbonate, ethyl acetate, benzyl alcohol, benzyl benzoate, propylene glycol, 1,3-butylene glycol, oils (in particular, cottonseed, groundnut, corn, germ, olive, castor, and sesame oils), glycerol, tetrahydrofuryl alcohol, polyethylene glycols and fatty acid esters of sorbitan, or mixtures thereof. Besides inert diluents, the oral compositions can also include adjuvants such as wetting agents, emulsifying and suspending agents, sweetening, flavoring, coloring, perfuming, and preservative agents. [00232] Compositions suitable for buccal or sublingual administration include tablets, lozenges and pastilles, wherein the active ingredient is formulated with a carrier such as sugar and acacia, tragacanth, or gelatin and glycerin. [00233] Solid compositions of a similar type may also be employed as fillers in soft and hard- filled gelatin capsules using excipients such as lactose or milk sugar, as well as high molecular weight polyethylene glycols and the like. The solid dosage forms of tablets, dragees, capsules, pills, and granules can be prepared with coatings and shells such as enteric coatings and other coatings well known in the pharmaceutical formulating art. They may optionally contain opacifying agents and can also be of a composition that they release the active ingredient(s) only, or preferentially, in a certain part of the intestinal tract, optionally, in a delayed manner. Examples of embedding compositions that can be used include polymeric substances and waxes. [00234] A therapeutic agent can also be in micro-encapsulated form with one or more excipients, as noted above. In such solid dosage forms, the therapeutic agent can be admixed with at least one inert diluent such as sucrose, lactose or starch. Such dosage forms can also comprise, as is normal practice, additional substances other than inert diluents, e.g., tableting lubricants and other tableting aids such a magnesium stearate and microcrystalline cellulose. [00235] Compositions for oral administration may be designed to protect the active ingredient against degradation as it passes through the alimentary tract, for example, by an outer coating of the formulation on a tablet or capsule. [00236] In another aspect, a composition can be an extended (or “delayed” or “sustained”) release composition. This delayed-release composition comprises a delayed-release component. Such a composition allows targeted release of a therapeutic agent, for example, into the lower gastrointestinal tract, for example, into the small intestine, the large intestine, the colon and/or the rectum. In certain aspects, a delayed-release composition further comprises an enteric or pH-dependent coating, such as cellulose acetate phthalates and other phthalates (e.g., polyvinyl acetate phthalate, methacrylates (Eudragits)). Alternatively, the delayed-release composition can provide controlled release to the small intestine and/or colon by the provision of pH sensitive methacrylate coatings, pH sensitive polymeric microspheres, or polymers which undergo degradation by hydrolysis. The delayed-release composition can be formulated with hydrophobic or gelling excipients or coatings. Colonic delivery can further be provided by coatings which are digested by bacterial enzymes such as amylose or pectin, by pH dependent polymers, by hydrogel plugs swelling with time (Pulsincap), by time-dependent hydrogel coatings and/or by acrylic acid linked to azoaromatic bonds coatings. [00237] Compositions described herein can also be administered subcutaneously, intraperitoneally or intravenously, e.g., in the form of a sterile injectable preparation, for example, as a sterile injectable aqueous or oleaginous suspension. This suspension can be formulated according to techniques known in the art using suitable dispersing or wetting agents (such as, for example, Tween 80) and suspending agents. The sterile injectable preparation can also be a sterile injectable solution or suspension in a non-toxic parenterally acceptable diluent or solvent, for example, as a solution in 1,3-butanediol. Among the acceptable vehicles and solvents that can be employed are mannitol, dextrose, water, Ringer’s solution, lactated Ringer’s solution and isotonic sodium chloride solution. In addition, sterile, fixed oils are conventionally employed as a solvent or suspending medium. For this purpose, any bland fixed oil can be employed including synthetic mono- or diglycerides. Fatty acids, such as oleic acid and its glyceride derivatives are useful in the preparation of injectables, as are natural pharmaceutically- acceptable oils, such as olive oil or castor oil, especially in their polyoxyethylated versions. These oil solutions or suspensions can also contain a long-chain alcohol diluent or dispersant, or carboxymethyl cellulose or similar dispersing agents which are commonly used in the formulation of pharmaceutically acceptable dosage forms such as emulsions and or suspensions. Other commonly used surfactants such as Tweens or Spans and/or other similar emulsifying agents or bioavailability enhancers which are commonly used in the manufacture of pharmaceutically acceptable solid, liquid, or other dosage forms can also be used for the purposes of formulation. [00238] Compositions described herein can also be administered in the form of suppositories for rectal administration. These can be prepared by mixing a compound or lipid particle described herein, e.g., in the form of a formulation of the disclosure, with a suitable non- irritating excipient that is solid at room temperature but liquid at rectal temperature and, therefore, will melt in the rectum to release the drug. Such materials include cocoa butter, beeswax and polyethylene glycols. [00239] Compositions described herein can also be administered topically, especially when the target of treatment includes areas or organs readily accessible by topical application, including diseases of the eye, the skin, or the lower intestinal tract. Suitable topical formulations are readily prepared for each of these areas or organs. [00240] Topical application for the lower intestinal tract can be effected in a rectal suppository formulation (see above) or in a suitable enema formulation. Topically-transdermal patches can also be used. [00241] For other topical applications, the compositions can be formulated in a suitable ointment containing the active component suspended or dissolved in one or more carriers. Carriers for topical administration include, but are not limited to, mineral oil, liquid petrolatum, white petrolatum, propylene glycol, polyoxyethylene, polyoxypropylene compound, emulsifying wax and water and penetration enhancers. Alternatively, compositions can be formulated in a suitable lotion or cream containing the active compound suspended or dissolved in one or more pharmaceutically acceptable carriers. Alternatively, the composition can be formulated with a suitable lotion or cream containing the active compound suspended or dissolved in a carrier with suitable emulsifying agents. Suitable carriers include, but are not limited to, mineral oil, sorbitan monostearate, polysorbate 60, cetyl esters wax, cetearyl alcohol, 2-octyldodecanol, benzyl alcohol and water. Suitable carriers also include, but are not limited to, mineral oil, sorbitan monostearate, polysorbate 60, cetyl esters wax, cetearyl alcohol, 2-octyldodecanol, benzyl alcohol and water and penetration enhancers. [00242] For ophthalmic use, compositions can be formulated as micronized suspensions in isotonic, pH adjusted sterile saline, or, preferably, as solutions in isotonic, pH adjusted sterile saline, either with or without a preservative such as benzylalkonium chloride. Alternatively, for ophthalmic use, the compositions can be formulated in an ointment such as petrolatum. [00243] Compositions can also be administered by nasal aerosol or inhalation, e.g., in the form of an aerosol formulation. Such compositions are prepared according to techniques well-known in the art of pharmaceutical formulation and can be prepared as solutions in saline, employing benzyl alcohol or other suitable preservatives, absorption promoters to enhance bioavailability, fluorocarbons, and/or other conventional solubilizing or dispersing agents. Aerosol formulations can be placed into pressurized acceptable propellants, such as dichlorodifluoromethane, propane, nitrogen, and the like. Without wishing to be bound by any particular theory, it is believed that local delivery of a composition described herein, as can be achieved by nasal aerosol or inhalation, for example, can reduce the risk of systemic consequences of the composition, for example, consequences for red blood cells. [00244] Other pharmaceutically acceptable carriers that can be used in the compositions described herein include, but are not limited to, ion exchangers, alumina, aluminum stearate, lecithin, self-emulsifying drug delivery systems (SEDDS) such as d-α-tocopherol polyethylene glycol 1000 succinate, surfactants used in pharmaceutical dosage forms such as Tweens or other similar polymeric delivery matrices, serum proteins, such as human serum albumin, buffer substances such as phosphates, glycine, sorbic acid, potassium sorbate, partial glyceride mixtures of saturated vegetable fatty acids, water, salts or electrolytes, such as protamine sulfate, disodium hydrogen phosphate, potassium hydrogen phosphate, sodium chloride, zinc salts, colloidal silica, magnesium trisilicate, polyvinyl pyrrolidone, cellulose-based substances, polyethylene glycol, sodium carboxymethylcellulose, polyacrylates, waxes, polyethylene- polyoxypropylene-block polymers, polyethylene glycol and wool fat. Cyclodextrins such as α-, β-, and γ-cyclodextrin, or chemically modified derivatives thereof, such as hydroxyalkylcyclodextrins, including hydroxylpropyl-β-cyclodextrins, such as 2- and/or 3- hydroxypropyl-β-cyclodextrins, or other solubilized derivatives thereof can also be advantageously used as a pharmaceutically acceptable carrier in the compositions described herein, e.g., to enhance delivery of agents described herein. [00245] In some aspects, a composition described herein includes one or more additional therapeutic agents, e.g., for use in combination with a therapeutic agent included in a lipid particle described herein. [00246] Some embodiments provide a combination (e.g., pharmaceutical combination) comprising a composition described herein comprising a compound of the disclosure, and one or more additional therapeutic agents (e.g., one or more compositions comprising one or more additional therapeutic agents). Such combinations are particularly useful as, for example, when a composition comprising a compound of the disclosure and the one or more therapeutic agents are to be administered separately. In a combination provided herein, the composition comprising a compound of the disclosure and the one or more therapeutic agents can be administrable by the same route of administration or by different routes of administration. [00247] The compositions described herein can be provided in unit dosage form. The amount of active ingredient in the composition of the disclosure that can be combined with a carrier to produce a unit dosage form will vary depending, for example, upon the subject being administered to and the particular mode of administration. Typically, a unit dosage form comprising a nucleic acid(s) active ingredient will contain from about 1 to about 1,000 µg of active ingredient(s), e.g., from about 1 to about 500 µg, from about 1 to about 250 µg, from about 1 to about 150 µg, from about 0.5 to about 100 µg, or from about 1 to about 50 µg of active ingredient(s). In some embodiments, a unit dosage form contains from about 0.01 µg to about 100 µg of active ingredient(s), e.g., from about 0.1 µg to about 50 µg, from about 0.1 µg to about 25 µg, from about 0.5 µg to about 50 µg of active ingredient(s). In some aspects, a unit dosage form contains from about 1 µg to about 5,000 µg of active ingredient(s) e.g., from about 10 µg to about 2,500 µg, from about 15 µg to about 1,000 µg or from about 100 µg to about 1,000 µg of active ingredient(s). In some aspects, a unit dosage form contains about 15 µg, about 30 µg, about 50 µg, about 100 µg, about 125 µg or about 150 µg of active ingredient(s). Methods of Use [00248] It has now been found that compounds of the disclosure, upon incorporation into a lipid nanoparticle and administration to a subject, are capable of delivering a nucleic acid encapsulated within the lipid particle to the subject. [00249] In certain embodiments, described herein is a method of administering a therapeutic agent and/or nucleic acid to a subject (e.g., a subject in need thereof), comprising administering to the subject a lipid nanoparticle described herein comprising the therapeutic agent and/or nucleic acid, e.g., a composition comprising a lipid nanoparticle described herein comprising the therapeutic agent and/or nucleic acid. Some embodiments comprise administering an effective amount of the lipid nanoparticle or composition to the subject, e.g., to treat a disease or condition in the subject. [00250] In certain embodiments, described herein is a method of treating a disease or condition in a subject in need thereof, comprising administering to the subject a lipid nanoparticle described herein comprising a therapeutic agent and/or nucleic acid, e.g., a therapeutic agent and/or nucleic acid known to treat the disease or condition. In some embodiments, a composition comprising the lipid nanoparticle is administered to the subject. Some embodiments comprise administering an effective amount of the lipid nanoparticle or composition to the subject. [00251] In some embodiments, the subject (e.g., subject in need thereof) is a mammal. In further embodiments, the subject (e.g., subject in need thereof) is a human. [00252] In some embodiments, a method described herein further comprises administering to the subject (e.g., an effective amount of) an additional therapy(ies), e.g., in combination with lipid nanoparticle described herein comprising a therapeutic agent and/or nucleic acid, e.g., a composition comprising a lipid nanoparticle described herein comprising a therapeutic agent and/or nucleic acid. In some embodiments, the lipid nanoparticle is administered before the additional therapy(ies). In some embodiments, the lipid nanoparticle is administered after the additional therapy(ies). In some aspects, the lipid nanoparticle is administered concurrently with the additional therapy(ies). [00253] An effective amount of a composition to be administered can be determined by a clinician of ordinary skill using the guidance provided herein and other methods known in the art. Preferably, the dosage causes or produces minimal adverse side effects. Specific dosage and treatment regimens for any particular patient will depend, for example, upon a variety of factors, such as the activity of the specific agent employed, the age, body weight, general health status, sex, diet, time of administration, rate of excretion, drug combination, the severity and course of the disease, condition or symptoms, the subject’s disposition to the disease, condition or symptoms, and the judgment of the treating physician. Determining the dosage for a particular agent, subject and disease, disorder or condition is within the abilities of one of skill in the art. [00254] A lipid nanoparticle or composition described herein can be administered via a variety of routes of administration, including, for example, oral, dietary, topical, transdermal, rectal, parenteral (e.g., intra-arterial, intravenous, intramuscular, subcutaneous injection, intradermal injection), intravenous infusion and inhalation (e.g., intrabronchial, intranasal or oral inhalation, intranasal drops) routes of administration, depending on the compound, antigen and/or therapeutic agent, respectively, and the particular disease to be treated. Administration can be local or systemic as indicated. The preferred mode of administration can vary depending on the particular compound or agent. [00255] Lipid nanoparticles described herein and compositions thereof are also useful for the introduction of therapeutic agents or nucleic acids into cells. The administration is carried out in vitro or in vivo by contacting the particles with the cells for a period of time sufficient for delivery of the therapeutic agent or nucleic acid agent to the cells to occur. In some aspects, the compounds and compositions described herein are administered in vitro. In some aspects, the compounds and compositions described herein are administered in vivo. [00256] For in vitro applications, the delivery of therapeutic agents or nucleic acids (e.g., interfering RNA or mRNA) can be to any cell grown in culture, whether of plant or animal origin, vertebrate or invertebrate, and of any tissue or type. In some embodiments, the cells are animal cells, preferably mammalian cells, and more preferably human cells. Contact between the cells and the lipid particles, when carried out in vitro, takes place in a biologically compatible medium. Treatment of the cells with the lipid particles may be carried out at physiological temperatures (e.g., about 37° C). In some embodiments, the treatment is for a period of time of from about 1 to about 48 hours, preferably of from about 2 to about 4 hours. EXAMPLES [00257] The following Examples are presented by way of illustration, not limitation. Compounds are named using the automatic name generating tool provided in Chemdraw Ultra 20.1 (Cambridgesoft), which generates systematic names for chemical structures, with support for the Cahn-Ingold-Prelog rules for stereochemistry. One skilled in the art can modify the procedures set forth in the illustrative examples to arrive at the desired products. Table of Abbreviations

Chemical Synthesis: Example 1: Synthesis of bis(2-butyloctyl) 10-[3-(dimethylamino)propyl- heptylsulfanylcarbonyl-amino]nonadecanedioate (1):

[00258] Step 1: tetraethyl 9-oxoheptadecane-1,8,10,17-tetracarboxylate (1c):

[00259] To a solution of NaOEt (84.14 g, 247.28 mmol, 20% purity, 1.0 eq) in EtOH (300 mL) was added diethyl 3-oxopentanedioate (1a, 50 g, 247.28 mmol, 44.88 mL, 1.0 eq). The stirred solution was heated to 80 °C, and then ethyl 8-bromooctanoate (1b, 62.11 g, 247.28 mmol, 1.0 eq) was added slowly. Stirring and heating continued for 2 hours. After another solution of NaOEt (84.14 g, 247.28 mmol, 20% purity, 1.0 eq) was added at 80 °C, another ethyl 8-bromooctanoate (62.11 g, 247.28 mmol, 1.0 eq) was added dropwise. The mixture was heated and stirred for 16 hours. Most of the ethanol was removed under reduced pressure. To the residue were added water (70 mL) and EtOAc (250 mL). Saturated NH₄C1 solution was added to adjust to pH = 7. The two layers were separated. The ethereal solution was washed again with solution of NH4C1 (100 mL × 2), brine (150 mL × 3), and dried over anhydrous sodium sulfate, filtered, and concentrated to give a crude product. The residue was purified by column chromatography (SiO₂, petroleum ether / EtOAc = 1 / 0 to 30 / 1) to give compound tetraethyl 9- oxoheptadecane-1,8,10,17-tetracarboxylate (1c, 60 g, 110.56 mmol, 46.2% yield) as a white solid. ¹H NMR (400 MHz, CDCl₃) δ = 4.27 - 4.12 (m, 8H), 3.61 - 3.57 (m, 2H), 2.26 (t, J = 7.6 Hz, 4H), 1.91 - 1.73 (m, 4H), 1.35 - 1.22 (m, 32H). [00260] Step 2: 10-oxononadecanedioic acid (1d):

[00261] To a solution of tetraethyl 9-oxoheptadecane-1,8,10,17-tetracarboxylate (1c, 30 g, 55.28 mmol, 1.0 eq) in HCl (60 mL) and HOAc (30 mL). The mixture was stirred at 115 °C for 5 hours. After the reaction was completed. The reaction mixture was cooled down to room temperature, a large quantity of an off-white precipitate was formed, and then the solid was filtered, the filtered cake was washed with petroleum ether / EtOAc (10 / 1, v/v, 200 mL) to give compound 10-oxononadecanedioic acid (1d, 5.5 g, 16.06 mmol, 29.1% yield) as a white solid. ¹H NMR (400 MHz, DMSO-d₆) δ = 11.94 (s, 2H), 2.37 (t, J = 7.2 Hz, 4H), 2.18 (t, J = 7.2 Hz, 4H), 1.56 - 1.36 (m, 8H), 1.34 - 1.14 (m, 16H). [00262] Step 3: bis(2-butyloctyl) 10-oxononadecanedioate (1f):

[00263] To a solution of 10-oxononadecanedioic acid (1d, 3 g, 8.76 mmol, 1.0 eq), 2- butyloctan-1-ol (1e, 3.59 g, 19.27 mmol, 2.2 eq), TEA (4.43 g, 43.80 mmol, 6.10 mL, 5.0 eq) and DMAP (3.21 g, 26.28 mmol, 3 eq) in DCM (40 mL) was added EDCI (4.20 g, 21.90 mmol, 2.5 eq). The mixture was stirred at 20 °C for 24 hours. The reaction mixture was partitioned between DCM (100 mL × 3) and H2O (150 mL). The organic phase was separated, washed with brine 150 mL (50 mL × 3), dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure to give a residue. The residue was purified by column chromatography (SiO2, petroleum ether / EtOAc = 1 / 0 to 50 / 1, v/v). then was further purified by flash silica gel chromatography (ISCO®; 120 g SepaFlash® Silica Flash Column, Eluent of 0 - 4% EtOAc / petroleum ether gradient at 100 mL / min) to give compound bis(2-butyloctyl) 10- oxononadecanedioate (1f, 3 g, 4.42 mmol, 42.9% yield) as a colorless oil. ¹H NMR (400 MHz, CDCl3) δ =3.97 (d, J = 5.6 Hz, 4H), 2.37 (t, J = 7.2 Hz, 4H), 2.29 (t, J = 7.6 Hz, 4H), 1.68 - 1.54 (m, 9H), 1.28 (s, 48H), 0.91 - 0.88 (m, 12H). [00264] Step 4: bis(2-butyloctyl) 10-((3-(dimethylamino)propyl)amino)nonadecanedioate (1h):

[00265] A mixture of N',N'-dimethylpropane-1,3-diamine (1g, 290 mg, 2.84 mmol, 354.96 μL, 1.0 eq), NaBH(OAc)3 (902.30 mg, 4.26 mmol, 1.5 eq), HOAc (255.66 mg, 4.26 mmol, 243.72 μL, 1.5 eq) in DCM (20 mL) was degassed and purged with N23 times, bis(2-butyloctyl) 10- oxononadecanedioate (1f, 2.31 g, 3.41 mmol, 1.2 eq) in DCM (1 mL) was added and then the mixture was stirred at 20 °C for 10 hours under N2. The mixture pH was adjusted to 9 - 10 with 1 N NaOH (20 mL), the aqueous phase was extracted with EtOAc 105 mL (35 mL × 3). The combined organic phase was washed with brine (40 mL × 2), dried with anhydrous sodium sulfate, filtered, and concentrated in vacuum to give crude product. The residue was purified by column chromatography (SiO2, DCM / MeOH = 1 / 0 to 30 / 1) to give compound bis(2- butyloctyl)10-[3-(dimethylamino)propylamino]nonadecanedioate (1h, 1.77 g, 2.08 mmol, 73.3% yield, 90.0% purity) as a yellow oil. ¹H NMR (400 MHz, CDCl₃) δ = 3.97 (d, J = 6.0 Hz, 4H), 2.65 (t, J = 6.4 Hz, 2H), 2.46 (d, J = 5.6 Hz, 1H), 2.35 - 2.26 (m, 6H), 2.23 (s, 6H), 1.73 - 1.51 (m, 9H), 1.37 - 1.15 (m, 56H), 0.96 - 0.80 (m, 12H). [00266] Step 5: bis(2-butyloctyl) 10-((3- (dimethylamino)propyl)((heptylthio)carbonyl)amino)nonadecanedioate (1):

[00267] To a solution of bis(2-butyloctyl) 10-[3- (dimethylamino)propylamino]nonadecanedioate (1h, 1.27 g, 1.66 mmol, 1.0 eq) and TEA (503.77 mg, 4.98 mmol, 692.95 μL, 3.0 eq) dissolved in dry DCM (15 mL) was added triphosgene (295.47 mg, 995.71 μmol, 0.6 eq) at 0 °C under N₂. The resulting solution was stirred at 20 °C for 1 hour (checked by TLC) and concentrated under reduced pressure and kept under N2. To a solution of heptane-1-thiol (1i, 439.00 mg, 3.32 mmol, 520.14 μL, 2.0 eq) dissolved in dry (10 mL) was added NaOH (331.88 mg, 8.30 mmol, 5.0 eq) at 0 °C under N₂. To this resulting solution the above carbamoyl chloride dissolved in THF (5 mL) was added via syringe slowly under N2 at 0 °C. The resulting solution was stirred at 25 ° C for 10 hours. The reaction mixture was quenched by NH₄Cl (60 mL) at 0 °C and then diluted with H₂O (30 mL). The aqueous phase was extracted with EtOAc (30 mL × 2). The combined organic phase was washed with brine (60 mL), dried with anhydrous sodium sulfate, filtered, and concentrated in vacuum to give residue. The residue was purified by flash silica gel chromatography (ISCO®; 20 g SepaFlash® Silica Flash Column, Eluent of 0 - 10% MeOH / DCM gradient at 80 mL / min) and prep-HPLC (Formic acid condition; column: C1250 × 80 mm, 10 μm; mobile phase: [water – ACN with formic acid]; gradient: 60% - 90% B over 10 min) to give compound bis(2- butyloctyl) 10-[3-(dimethylamino)propyl-heptylsulfanylcarbonyl-amino]nonadecanedioate (1, 328 mg, 355.12 μmol, 21.4% yield, 99.99% purity) as a yellow oil. LCMS: [M+H]⁺: 924.0; ¹H NMR (400 MHz, CDCl3) δ = 3.97 (d, J = 5.6 Hz, 4H), 3.78 (s, 1H), 3.22 - 3.10 (m, 2H), 2.88 (t, J = 7.2 Hz, 2H), 2.39 - 2.22 (m, 12H), 1.85 - 1.73 (m, 2H), 1.67 - 1.55 (m, 8H), 1.51 - 1.41 (m, 4H), 1.37 - 1.16 (m, 60H), 0.99 - 0.79 (m, 15H). Example 2: Synthesis of bis(2-butyloctyl) 10-[heptylsulfanylcarbonyl-(1-methyl-4- piperidyl)amino]nonadecanedioate (2):

[00268] Step 1: bis(2-butyloctyl) 10-((1-methylpiperidin-4-yl)amino)nonadecanedioate (2b):

[00269] A mixture of 1-methylpiperidin-4-amine (980 mg, 8.58 mmol, 1.0 eq), NaBH(OAc)3 (2.73 g, 12.87 mmol, 1.5 eq), HOAc (773.08 mg, 12.87 mmol, 736.97 μL, 1.5 eq) in DCM (70 mL) was degassed and purged with N₂3 times, bis(2-butyloctyl) 10-oxononadecanedioate (6.99 g, 10.30 mmol, 1.2 eq) in DCM (10 mL) was added and then the mixture was stirred at 20 °C for 10 hours under N2. The mixture pH was adjusted to 9 - 10 with 1 N NaOH (20 mL), the aqueous phase was extracted with EtOAc (35 mL × 3). The combined organic phase was washed with brine (40 mL), dried with anhydrous sodium sulfate, filtered, and concentrated in vacuum to give residue. The residue was purified by column chromatography (SiO2, petroleum ether / EtOAc = 1/0 to 0/1, v/v) to give compound bis(2-butyloctyl) 10-[(1-methyl-4- piperidyl)amino]nonadecanedioate (2b, 2.2 g, 2.55 mmol, 29.7% yield, 90% purity) as a yellow oil. ¹H NMR (400 MHz, CDCl3) δ = 3.96 (d, J = 5.6 Hz, 4H), 2.85 (d, J = 11.2 Hz, 2H), 2.55 (d, J = 5.6 Hz, 2H), 2.39 - 2.19 (m, 8H), 2.16 - 2.20 (m, 2H), 1.89 (d, J = 12.0 Hz, 2H), 1.65 - 1.56 (m, 6H), 1.46 - 1.40 (m, 2H), 1.37 - 1.10 (m, 58H), 1.04 - 0.72 (m, 12H). [00270] Step 2: bis(2-butyloctyl) 10-(((heptylthio)carbonyl)(1-methylpiperidin-4- yl)amino)nonadecanedioate (2):

[00271] To a solution of bis(2-butyloctyl) 10-[(1-methyl-4-piperidyl)amino]nonadecanedioate (2b, 2.05 g, 2.64 mmol, 1.0 eq) and TEA (800.61 mg, 7.91 mmol, 1.10 mL, 3.0 eq) dissolved in dry DCM (20 mL) was added triphosgene (469.58 mg, 1.58 mmol, 0.6 eq) at 0 °C under N2. The resulting solution was stirred at 20 °C for 1 hour (checked by TLC) and concentrated under reduced pressure and kept under N₂. To a solution of heptane-1-thiol (1i, 697.67 mg, 5.27 mmol, 826.62 μL, 2.0 eq) dissolved in dry THF (20 mL) was added NaOH (527.43 mg, 13.19 mmol, 5.0 eq) at 0 °C under N2. To this resulting solution the above carbamoyl chloride, dissolved in THF (10 mL) was added via syringe slowly under N₂ at 0 °C. The resulting solution was stirred at 25 ° C for 15 hours. The reaction mixture was quenched by NH4Cl (20 mL) at 0°C and then diluted with H2O (40 mL). The aqueous phase was extracted with DCM (30 mL × 2). The combined organic phase was washed with brine (30 mL), dried with anhydrous sodium sulfate, filtered, and concentrated in vacuum to give residue. The residue was purified by flash silica gel chromatography (ISCO®; 40 g SepaFlash® Silica Flash Column, Eluent of 0 - 10% MeOH / DCM gradient at 100 mL/min) and prep-HPLC (Formic acid condition; column: C1250 × 80 mm, 10 μm; mobile phase: [water (formic acid) - ACN]; gradient: 60% - 90% B over 10 min) to give compound bis(2-butyloctyl) 10-[heptylsulfanylcarbonyl-(1-methyl-4- piperidyl)amino]nonadecanedioate (2, 771 mg, 824.03 μmol, 31.2% yield, 99.99% purity) as a yellow oil. LCMS: [M+H]⁺: 936.2; ¹H NMR (400 MHz, CD₃OD) δ = 4.00 (d, J = 5.6 Hz, 4H), 3.90 (s, 1H), 3.47 - 3.30 (m, 2H), 2.99 - 2.81 (m, 4H), 2.70 - 2.47 (m, 4H), 2.31 (t, J = 7.2 Hz, 4H), 1.66 - 1.54 (m, 12H), 1.46 - 1.23 (m, 64H), 0.98 - 0.85 (m, 15H). Example 3: Synthesis of bis(2-butyloctyl) 10-[3-(dimethylamino)propyl-octylsulfonyl- amino]nonadecanedioate (3):

[00272] Step 1: octane-1-sulfonyl chloride (3a): N_{CS (3.5 eq.), IPA}

[00273] To a solution of octane-1-thiol (5 g, 34.18 mmol, 5.93 mL, 1.0 eq) and propan-2-ol (4.11 g, 68.36 mmol, 5.23 mL, 2.0 eq) in DCM (70 mL) was added NCS (15.97 g, 119.62 mmol, 3.5 eq) at 0 °C. The mixture was stirred at 25 °C for 2 hours. The reaction mixture was diluted with NaHCO330 mL at 0 °C and extracted with EtOAc 60 mL (30 mL × 2). The combined organic layers were washed with brine 60 mL (20 mL × 3), dried over sodium sulfate, filtered, and concentrated under reduced pressure to give a residue. The residue was purified by flash silica gel chromatography (ISCO®; 80 g SepaFlash® Silica Flash Column, Eluent of 0 - 0% EtOAc/petroleum ether at 100 mL/min) to give compound octane-1-sulfonyl chloride (3a, 6.3 g, 29.61 mmol, 86.7% yield) as a yellow oil. ¹H NMR (400 MHz, CDCl3) δ = 3.73 - 3.59 (m, 2H), 2.12 - 1.97 (m, 2H), 1.50 (q, J = 7.2 Hz, 2H), 1.39 - 1.27 (m, 8H), 0.94 - 0.86 (m, 3H). [00274] Step 2: bis(2-butyloctyl) 10-(N-(3- (dimethylamino)propyl)octylsulfonamido)nonadecanedioate (3):

[00275] To a solution of bis(2-butyloctyl) 10-[3- (dimethylamino)propylamino]nonadecanedioate (1h, 2.46 g, 3.21 mmol, 1.0 eq), DMAP (78.54 mg, 642.90 μmol, 0.2 eq) and TEA (1.63 g, 16.07 mmol, 2.24 mL, 5.0 eq) in DCM (36 mL) was added octane-1-sulfonyl chloride (2.74 g, 12.86 mmol, 4.0 eq) at 0 °C. The mixture was stirred at 25 °C for 16 hours. The reaction mixture was partitioned between DCM (60 mL) and H2O (50 mL). The organic phase was separated, washed with brine 70 mL (35 mL × 2), dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure to give a residue. The residue was purified by flash silica gel chromatography (ISCO®; 20 g SepaFlash® Silica Flash Column, Eluent of 0 - 10% EtOAc / petroleum ether at 80 mL / min) and prep-HPLC (Formic Acid condition; column: C1250 × 80 mm, 10 μm; mobile phase: [water (Formic Acid) - ACN]; gradient: 54% - 84% B over 10 min) to give compound bis(2-butyloctyl) 10-[3- (dimethylamino)propyl-octylsulfonyl-amino]nonadecanedioate (3, 720 mg, 749.17 μmol, 23.3% yield, 97.97% purity) as a yellow oil. LCMS: [M+H]⁺: 942.0; ¹H NMR (400 MHz, CDCl3) δ = 3.97 (d, J = 6.0 Hz, 4H), 3.60 - 3.46 (m, 1H), 3.17 - 3.08 (m, 2H), 2.93 - 2.83 (m, 2H), 2.40 - 2.33 (m, 2H), 2.32 - 2.29 (m, 4H), 2.28 (s, 6H), 1.86 - 1.79 (m, 4H), 1.63 - 1.59 (m, 4H), 1.53 - 1.44 (m, 4H), 1.43 - 1.36 (m, 4H), 1.33 - 1.25 (m, 60H), 0.91 - 0.86 (m, 15H). Example 4: Synthesis of bis(1-hexylheptyl) 10-[3-(dimethylamino)propyl- hexylsulfanylcarbonyl-amino]nonadecanedioate (4):

[00276] Step 1: bis(1-hexylheptyl) 10-oxononadecanedioate (4b):

1d 4b [00277] To a solution of 10-oxononadecanedioic acid (1d, 4 g, 11.68 mmol, 1 eq), tridecan-7- ol (4a, 5.15 g, 25.70 mmol, 2.2 eq), TEA (5.91 g, 58.40 mmol, 8.13 mL, 5 eq) and DMAP (4.28 g, 35.04 mmol, 3 eq) in DCM (40 mL) was added EDCI (5.60 g, 29.20 mmol, 2.5 eq). The mixture was stirred at 20 °C for 24 hours. The reaction mixture was partitioned between DCM 60 mL and H2O 80 mL. The organic phase was separated, washed with brine 100 mL (50 mL x 2), dried over Na₂SO₄, filtered, and concentrated under reduced pressure to give a residue. The residue was purified by flash silica gel chromatography (120 g SepaFlash® Silica Flash Column, petroleum ether : EtOAc: 0-10%) to give compound bis(1-hexylheptyl) 10-oxononadecanedioate (4b, 6 g, 8.48 mmol, 72.6% yield) as yellow oil. [00278] Step 2: bis(1-hexylheptyl) 10-[3-(dimethylamino)propylamino]nonadecanedioate (4d):

[00279] A mixture of N',N'-dimethylpropane-1,3-diamine (4c, 866.94 mg, 8.48 mmol, 1.06 mL, 1.2 eq) and NaBH(OAc)₃ (2.25 g, 10.61 mmol, 1.5 eq), HOAc (636.88 mg, 10.61 mmol, 607.13 μL, 1.5 eq) in DCM (60 mL) was stirred at 25 °C for 1 hour, and then bis(1-hexylheptyl) 10-oxononadecanedioate (4b, 5 g, 7.07 mmol, 1 eq) was added. The resulting mixture was stirred at 25 °C for 15 hours. The reaction mixture was diluted with H₂O (50 mL) and the mixture pH was adjusted to 9-10 with saturated Na2CO3, the aqueous phase was extracted with DCM (50 mL x 3). The combined organic phase was washed with brine (50 mL), dried with anhydrous Na₂SO₄, filtered and concentrated under reduced pressure to give a residue. The residue was purified by flash silica gel chromatography (80 g SepaFlash® Silica Flash Column, DCM : MeOH: 0-10%) to give compound bis(1-hexylheptyl) 10-[3- (dimethylamino)propylamino]nonadecanedioate (4d, 4.1 g, 5.17 mmol, 73.2% yield) as yellow oil. ¹H NMR (400 MHz, CDCl₃) δ = 4.90 - 4.85 (m, 2H), 2.75 - 2.65 (m, 2H), 2.55 - 2.45 (m, 1H), 2.36 (t, J = 6.8 Hz, 2H), 2.28 (t, J = 7.6 Hz, 4H), 2.24 (s, 6H), 1.72- 1.68 (m, 2H), 1.64 - 1.63 (m, 1H), 1.61 - 1.55 (m, 2H), 1.54 - 1.47 (m, 8H), 1.45 - 1.38 (m, 4H), 1.34 - 1.22 (m, 53H), 0.92 - 0.84 (m, 12H). [00280] Step 3: bis(1-hexylheptyl) 10-[3-(dimethylamino)propyl-hexylsulfanylcarbonyl- amino]nonadecanedioate (4):

[00281] To a solution of bis(1-hexylheptyl) 10-[3- (dimethylamino)propylamino]nonadecanedioate (4d, 2.5 g, 3.15 mmol, 1 eq) and TEA (956.62 mg, 9.45 mmol, 1.32 mL, 3 eq) dissolved in dry DCM (30 mL) was added triphosgene (561.08 mg, 1.89 mmol, 0.6 eq) at 0 °C under N₂. The resulting solution was stirred at 20 °C for 1 hour (checked by TLC) and concentrated under reduced pressure and kept under N2. To a solution of hexane-1-thiol (1i, 1.12 g, 9.45 mmol, 1.33 mL, 3 eq) dissolved in dry THF (20 mL) was added NaOH (756.30 mg, 18.91 mmol, 6 eq) at 0 °C under N₂. To this resulting solution the above carbamoyl chloride, dissolved in THF (20 mL) was added via syringe slowly under N2 at 0 °C. The resulting solution was stirred at 25 ° C for 15 hours. The reaction mixture was quenched by NH₄Cl (60 mL) at 0 °C and then diluted with DCM (50 mL). The aqueous phase was extracted with DCM (50 mL × 3). The combined organic phase was washed with brine (60 mL), dried with anhydrous Na2SO4, filtered, and concentrated in vacuum to give residue. The residue was purified by flash silica gel chromatography (40 g SepaFlash® Silica Flash Column, DCM : MeOH: 0-10%) and prep-HPLC (column: C1250 × 80 mm, 10 μm; mobile phase: [water (Formic Acid)-ACN]; gradient: 54%-84% B over 15 min) to give compound bis(1-hexylheptyl) 10-[3-(dimethylamino)propyl-hexylsulfanylcarbonyl-amino]nonadecanedioate (4, 422 mg, 445.33 μmol, 17.4% yield, 98.94% purity). LCMS: [M+H⁺]: 937.9; ¹H NMR (400 MHz, CDCl₃) δ = 4.91 - 4.83 (m, 2H), 3.80 - 3.68 (m, 1H), 3.25 - 3.11 (m, 2H), 2.89 (t, J = 7.6 Hz, 2H), 2.39 2.33 (m, 4H), 2.28 (t, J = 7.2 Hz, 6H), 1.88 - 1.76 (m, 4H), 1.67 - 1.55 (m, 8H), 1.54 - 1.43 (m, 12H), 1.32 - 1.23 (m, 56H), 0.91 - 0.85 (m, 12H). Example 5: Synthesis of bis(2-butyloctyl) 10-(((heptylthio)carbonyl)((1-methylpiperidin-4- yl)methyl)amino)nonadecanedioate (5):

[00282] Step 1: methyl 9-bromononanoate (5b):

[00283] A mixture of 9-bromononanoic acid (5a, 100 g, 421.70 mmol, 1 eq) in MeOH (1200 mL) was degassed and purged with N₂3 times, then H₂SO₄ (330.88 g, 3.37 mol, 179.83 mL, 8 eq) was added slowly at 0 °C, and then the mixture was stirred at 80 °C for 5 hours under N₂. After the reaction, returning to room temperature, NaHCO3 was added to neutralize the reaction solution adjust to pH = 7, the solvent was distilled off under reduced pressure, diluted with EtOAc (500 mL), and washed with H₂O (300 mL).The washed water was extracted once with EtOAc (500 mL × 3), the organic layer was dried over anhydrous sodium sulfate, and the solvent was evaporated under reduced pressure to give compound methyl 9-bromononanoate (5b, 94 g, crude) as a colorless oil. ¹H NMR (400 MHz, CDCl₃-d) δ = 3.67 (s, 3H), 3.45 - 3.34 (m, 2H), 2.30 (t, J = 7.6 Hz, 2H), 1.91 -1.77 (m, 2H), 1.68 - 1.57 (m, 2H), 1.51 - 1.37 (m, 2H), 1.34 - 1.26 (m, 6H). [00284] Step 2: dimethyl 10-cyano-10-tosylnonadecanedioate (5c):

[00285] To a 2000 mL three neck round bottom flask under N₂, was added DMSO (800 mL), followed by the addition of NaH (17.93 g, 448.17 mmol, 60% purity, 2.5 eq) in portions over a period of 0.5 hours. The mixture was allowed to stir for 0.5 hours at 25 °C following the addition, then 1-(isocyanomethylsulfonyl)-4-methyl-benzene (35 g, 179.27 mmol, 1.0 eq) was added in portions over 0.5 hours, followed by the addition of TBAI (6.62 g, 17.93 mmol, 0.1 eq) in one portion. The resulting mixture was stirred for 0.25 hours at 25 °C, then methyl 9- bromononanoate (5b, 94 g, 374.26 mmol, 2.1 eq) was added over a period of 0.5 hours. The mixture was stirred at 25 °C for 1 hour. The reaction mixture was quenched by addition water (300 mL) at 0 °C, and then extracted with petroleum ether / Ethyl Acetate = 10 / 1 (150 mL × 2). The combined organic layers were washed with brine (200 mL), dried over anhydrous sodium sulfate, filtered and concentrated under reduced pressure to give compound dimethyl 10-cyano- 10-(p-tolylsulfonyl)nonadecanedioate (5c) (68 g, 126.93 mmol, 70.8% yield) as a brown yellow oil. [00286] Step 3: dimethyl 10-oxononadecanedioate (5d):

[00287] To a 1000 mL three neck round bottom flask was added DCM (340 mL), followed by the addition of dimethyl 10-cyano-10-(p-tolylsulfonyl)nonadecanedioate (5c) (68.00 g, 126.93 mmol, 1.0 eq), under nitrogen, HCl (12 M, 85 mL, 8.0 eq) was added in portions over a period of 1 hour at 0 °C. The two-phase mixture was stirred at 25 °C for 12 hours. The reaction mixture was quenched by addition water (150 mL) at 0 °C, and then extracted with petroleum ether (150 mL × 2). The combined organic layers were washed with brine (200 mL), dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure to give residue. The residue was purified by column chromatography (SiO₂, Petroleum ether / Ethyl acetate = 1 / 0 to 20 / 1) to give compound dimethyl 10-oxononadecanedioate (5d) (19 g, 43.59 mmol, 34.4% yield, 85.0% purity) as a white solid. ¹H NMR (400 MHz, CDCl₃) δ = 3.65 (s, 6H), 2.37 (t, J = 7.2 Hz, 4H), 2.29 (t, J = 7.6 Hz, 4H), 1.63 - 1.51 (m, 8H), 1.36 - 1.26 (m, 16H). [00288] Step 4: 10-oxononadecanedioic acid (1d):

[00289] To a 250 mL three neck round bottom flask was added EtOH (240 mL) and H₂O (60 mL), followed by the addition of dimethyl 10-oxononadecanedioate (5d) (19 g, 51.28 mmol, 1.0 eq), under nitrogen, NaOH (6.15 g, 153.84 mmol, 3.0 eq) was added in portions over a period of 1 hour. The two-phase mixture was stirred at 25 °C for 16 hours. The reaction mixture was concentrated, diluted with H2O (100 mL), and then extracted with EtOAc (150 mL × 2). The aqueous phase was adjusted to pH = 3 with 1N HCl then filtered, and the filtered cake concentrated under reduced pressure to give compound 10-oxononadecanedioic acid (1d) (16.83 g, 46.69 mmol, 94.1% yield, 95.0% purity) as a white solid. ¹H NMR (400 MHz, DMSO-d₆) δ = 12.99 - 11.05 (m, 2H), 2.44 - 2.28 (m, 4H), 2.44 - 2.09 (m, 4H), 1.55 - 1.36 (m, 8H), 1.33 - 1.12 (m, 16H). [00290] Step 5: bis(2-butyloctyl) 10-(((1-methylpiperidin-4- yl)methyl)amino)nonadecanedioate (5f):

[00291] A mixture of (1-methyl-4-piperidyl)methanamine (5e) (900 mg, 7.02 mmol, 1.0 eq), NaBH(OAc)3 (2.22 g, 10.48 mmol, 1.5 eq), HOAc (629.61 mg, 10.48 mmol, 600.20 μL, 1.5 eq) in DCM (40 mL) was degassed and purged with N₂3 times, bis(2-butyloctyl) 10- oxononadecanedioate (1f) (5.70 g, 8.39 mmol, 1.2 eq) in DCM (10 mL) was added and then the mixture was stirred at 25 °C for 16 hours under N2. The mixture pH was adjusted to 9 - 10 was with 1 N NaOH (20 mL), the aqueous phase was extracted with EtOAc (35 mL × 3). The combined organic phase was washed with brine (40 mL), dried with anhydrous sodium sulfate, filtered, and concentrated in vacuum to give crude product. The residue was purified by flash silica gel chromatography (ISCO®; 80 g SepaFlash® Silica Flash Column, Eluent of 0 - 10% MeOH / DCM gradient at 100 mL / min) to give compound bis(2-butyloctyl) 10-[(1-methyl-4- piperidyl)methylamino]nonadecanedioate (5f) (3.2 g, 3.92 mmol, 56.1% yield, 97.0% purity) as a yellow oil. ¹H NMR (400 MHz, CDCl₃) δ = 3.97 (d, J = 5.6 Hz, 4H), 2.95 (d, J = 10.8 Hz, 2H), 2.52 - 2.40 (m, 3H), 2.37 - 2.24 (m, 7H), 2.12 - 1.98 (m, 2H), 1.84 - 1.73 (m, 2H), 1.67 - 1.57 (m, 6H), 1.41 - 1.21 (m, 60H), 0.99 - 0.81 (m, 12H). [00292] Step 6: bis(2-butyloctyl) 10-(((heptylthio)carbonyl)((1-methylpiperidin-4- yl)methyl)amino)nonadecanedioate (5):

[00293] To a solution of bis(2-butyloctyl) 10-[(1-methyl-4- piperidyl)methylamino]nonadecanedioate (5f) (2 g, 2.53 mmol, 1.0 eq) and TEA (767.25 mg, 7.58 mmol, 1.06 mL, 3.0 eq) dissolved in dry DCM (30 mL) was added triphosgene (450.01 mg, 1.52 mmol, 0.6 eq) at 0 °C under N₂. The resulting solution was stirred at 25 °C for 1 hour (checked by TLC) and concentrated under reduced pressure and kept under N2. To a solution of heptane-1-thiol (1.00 g, 7.58 mmol, 1.19 mL, 3.0 eq) dissolved in dry THF (25 mL) was added NaOH (303.29 mg, 7.58 mmol, 3.0 eq) at 0 °C under N₂. To this resulting solution the above carbamoyl chloride, dissolved in THF (20 mL) was added via syringe slowly under N2 at 0 °C. The resulting solution was stirred at 25 ° C for 15 hours. The reaction mixture was quenched by NH4Cl (16 mL) at 0 °C and then diluted with DCM (30 mL). The aqueous phase was extracted with EtOAc (30 mL × 2). The combined organic phase was washed with brine (60 mL), dried with anhydrous sodium sulfate, filtered, and concentrated in vacuum to give residue. The residue was purified by flash silica gel chromatography (ISCO®; 20 g SepaFlash® Silica Flash Column, Eluent of 0 - 10% MeOH / DCM gradient at 80 mL/min). then was further purified by prep- HPLC (Formic Acid condition; column: Phenomenex luna C18150 × 25 mm × 10 um; mobile phase: [water (Formic Acid) - ACN]; gradient: 60% - 90% B over 10 min) to give compound bis(2-butyloctyl) 10-[heptylsulfanylcarbonyl-[(1-methyl-4- piperidyl)methyl]amino]nonadecanedioate (5, 530 mg, 558.09 μmol, 33.1% yield, 99.99% purity) as a yellow oil. LCMS: [M+H]⁺: 950.0; ¹H NMR (400 MHz, CDCl₃) δ =3.97 (d, J = 6.0 Hz, 4H), 3.87 - 3.74 (m, 1H), 3.39 - 3.21 (m, 2H), 3.19 - 2.91 (m, 2H), 2.87 (t, J = 7.2 Hz, 2H), 2.55 (s, 3H), 2.52 - 2.28 (m, 2H), 2.30 (t, J = 7.6 Hz, 4H), 2.10 - 1.93 (m, 1H), 1.85 - 1.71 (m, 2H), 1.65 - 1.55 (m, 10H), 1.45 - 1.37 (m, 4H), 1.35 - 1.21 (m, 60H), 0.97 - 0.82 (m, 15H). Example 6: Synthesis of 8-(((heptylthio)carbonyl)(1-methylpiperidin-4-yl)amino)pentadecane- 1,15-diyl bis(4,4-bis(pentyloxy)butanoate) (6):

[00294] Step 1: 4,4-bis(pentyloxy)butanenitrile (6b):

[00295] A mixture of 4,4-diethoxybutanenitrile (6a) (3.5 g, 22.26 mmol, 1.0 eq) and PPTS (559.48 mg, 2.23 mmol, 0.1 eq) was dissolved in pentan-1-ol (5.89 g, 66.79 mmol, 7.26 mL, 3.0 eq) in a 50 mL single-necked round bottom flask. The mixture was stirred reflux at 110 °C for 16 hours under N₂. The residue was purified by flash silica gel chromatography (40 g SepaFlash® Silica Flash Column, petroleum ether : EtOAc: 0 - 10%) to give compound 4,4- dipentoxybutanenitrile (6b) (6.5 g, 26.93 mmol, 64.4% yield) as colorless oil.¹H NMR (400 MHz, CDCl₃) δ = 4.56 (t, J = 5.2 Hz, 1H), 3.65 - 3.55 (m, 2H), 3.48 - 3.40 (m, 2H), 2.50 - 2.36 (m, 2H), 2.02 - 1.90 (m, 2H), 1.65 - 1.56 (m, 4H), 1.42 - 1.30 (m, 8H), 0.97 - 0.85 (m, 6H). [00296] Step 2: 4,4-bis(pentyloxy)butanoic acid (6c):

[00297] A mixture of 4,4-dipentoxybutanenitrile (6b) (6.5 g, 26.93 mmol, 1.0 eq) and KOH (4.53 g, 80.79 mmol, 3.0 eq) in EtOH (35 mL) and H₂O (35 mL) was degassed and purged with N₂3 times, and then the mixture was stirred at 110 °C for 18 hours under N₂. The reaction mixture cooled down to room temperature, diluted with EtOAc (50 mL), pH was adjusted to pH = 5 by addition 1N HCl, and extracted with EtOAc (35 mL × 2). The combined organic layers were washed with brine (80 mL), dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure to give a residue. The residue was purified by flash silica gel chromatography (ISCO®; 80 g SepaFlash® Silica Flash Column, Eluent of 0 - 30% EtOAc / petroleum ether gradient at 100 mL / min) to give compound 4,4-dipentoxybutanoic acid (6c) (7 g, 24.20 mmol, 89.9% yield, 90.0% purity) as a yellow oil. ¹H NMR (400 MHz, DMSO-d6) δ = 12.03 (s, 1H), 4.45 (t, J = 5.6 Hz, 1H), 3.57 - 3.42 (m, 2H), 3.33 -3.24 (m, 2H), 2.22 (t, J = 7.2 Hz, 2H), 1.78 - 1.67 (m, 2H), 1.55 - 1.42 (m, 4H), 1.37 - 1.21 (m, 8H), 0.93 - 0.76 (m, 6H). [00298] Step 3: dimethyl 8-isocyano-8-tosylpentadecanedioate (6d):

[00299] To a 2000 mL three neck round bottom flask under N2, was added DMSO (500 mL), followed by the addition of NaH (10.24 g, 256.10 mmol, 60% purity, 2.5 eq) in portions over a period of 0.5 hours. The mixture was allowed to stir for 0.5 hours at 25 °C following the addition, then 1-(isocyanomethylsulfonyl)-4-methyl-benzene (20 g, 102.44 mmol, 1.0 eq) was added in portions over 0.5 hours, followed by the addition of TBAI (3.78 g, 10.24 mmol, 0.1 eq) in one portion. The resulting mixture was stirred for 0.5 hours at 25 °C, then methyl 7- bromoheptanoate (48.00 g, 215.12 mmol, 2.1 eq) was added over a period of 0.5 hours. The mixture was stirred at 25 °C for 1 hour. The reaction mixture was quenched by addition water (150 mL) at 0 °C, and then extracted with petroleum ether / EtOAc = 10 / 1 (150 mL × 2). The combined organic layers were washed with brine (200 mL), dried over anhydrous sodium sulfate, filtered and concentrated under reduced pressure to give compound dimethyl 8- isocyano-8-(p-tolylsulfonyl)pentadecanedioate (6d) (49.13 g, crude) as a brown oil. [00300] Step 4: dimethyl 8-oxopentadecanedioate (6e):

[00301] To a 1000 mL three neck round bottom flask was added DCM (240 mL), followed by the addition of dimethyl 8-isocyano-8-(p-tolylsulfonyl)pentadecanedioate (6d) (49.13 g, 102.43 mmol, 1.0 eq) , under nitrogen, HCl (12 M, 59.75 mL, 7.0 eq) was added in portions over a period of 1 hour. The two-phase mixture was stirred for 12 hours at 25 °C. The reaction mixture was quenched by addition water (150 mL) at 0 °C, and then extracted with petroleum ether (150 mL × 2). The combined organic layers were washed with brine (200 mL), dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure to give residue. The residue was purified by column chromatography (SiO2, petroleum ether / EtOAc = 1 / 0 to 20 / 1) to give compound dimethyl 8-oxopentadecanedioate (6e) (8.7 g, 23.52 mmol, 23.0% yield, 85.0% purity) as a colorless oil. ¹H NMR (400 MHz, CDCl₃) δ = 3.67 (s, 6H), 2.41 - 2.36 (m, 4H), 2.30 (t, J = 7.6 Hz, 4H), 1.65 - 1.54 (m, 8H), 1.36 - 1.28 (m, 8H). [00302] Step 5: dimethyl 8-((1-methylpiperidin-4-yl)amino)pentadecanedioate (6f):

[00303] A mixture of 1-methylpiperidin-4-amine (2.7 g, 23.65 mmol, 1.0 eq), NaBH(OAc)3 (7.52 g, 35.47 mmol, 1.5 eq), HOAc (2.13 g, 35.47 mmol, 2.03 mL, 1.5 eq) in DCM (90 mL) was degassed and purged with N₂3 times, dimethyl 8-oxopentadecanedioate (6e) (8.7 g, 27.67 mmol, 1.17 eq) in DCM (20 mL) was added and then the mixture was stirred at 25 °C for 10 hours under N2. The mixture pH was adjusted to 9-10 was with 1 M NaOH (20 mL), the aqueous phase was extracted with EtOAc (35 mL × 3). The combined organic phase was washed with brine (40 mL), dried with anhydrous sodium sulfate, filtered, and concentrated in vacuum to give crude product. The residue was purified by column chromatography (SiO2, DCM / MeOH = 1 / 0 to 30 / 1) to give compound dimethyl 8-[(1-methyl-4- piperidyl)amino]pentadecanedioate (6f) (9 g, 18.54 mmol, 52.3% yield, 85.0% purity) as a yellow oil. ¹H NMR (400 MHz, CDCl3) δ = 3.64 (s, 6H), 2.89 - 2.76 (m, 2H), 2.62 - 2.57 (m, 2H), 2.31 - 2.24 (m, 7H), 2.14 - 2.01 (m, 2H), 1.91 - 1.82 (m, 2H), 1.64 - 1.55 (m, 4H), 1.38 - 1.19 (m, 18H). [00304] Step 6: dimethyl 8-(((benzyloxy)carbonyl)(1-methylpiperidin-4- yl)amino)pentadecanedioate (6g):

[00305] A mixture of dimethyl 8-[(1-methyl-4-piperidyl)amino]pentadecanedioate (6f) (5.8 g, 14.06 mmol, 1.0 eq), DIEA (3.63 g, 28.11 mmol, 4.90 mL, 2.0 eq) in DCM (90 mL) was degassed and purged with N₂3 times, then benzyl (2,5-dioxopyrrolidin-1-yl) carbonate (4.20 g, 16.87 mmol, 1.2 eq) was added at 0 °C, and the reaction was stirred at 25 °C for 3 hours. The reaction mixture was concentrated under reduced pressure to give a residue. The residue was purified by flash silica gel chromatography (ISCO®; 80 g SepaFlash® Silica Flash Column, Eluent of 0 - 5% MeOH / DCM gradient at 100 mL / min) to give compound dimethyl 8- [benzyloxycarbonyl-(1-methyl-4-piperidyl)amino]pentadecanedioate (6g) (5.01 g, 8.25 mmol, 58.7% yield, 90.0% purity) as a yellow oil.¹H NMR (400 MHz, MeOD-d₄) δ = 7.43 - 7.22 (m, 5H), 5.26 - 5.01 (m, 2H), 4.26 - 3.99 (m, 1H), 3.65 (s, 6H), 3.01 - 2.73 (m, 3H), 2.55 - 2.24 (m, 9H), 2.09 (t, J = 10.4 Hz, 2H), 1.60 - 1.47 (m, 8H), 1.40 - 1.04 (m, 14H). [00306] Step 7: benzyl (1,15-dihydroxypentadecan-8-yl) (1-methylpiperidin-4-yl)carbamate

[00307] A mixture of dimethyl 8-[benzyloxycarbonyl-(1-methyl-4- piperidyl)amino]pentadecanedioate (6g) (5.01 g, 9.16 mmol, 1.0 eq) in THF (60 mL) was degassed and purged with N23 times and cooled down to 0 °C, then LiAlH4 (2.5 M, 7.70 mL, 2.1 eq) was added via injection at 0 °C, and then the mixture was stirred at 25 °C for 1.5 hours under N2. After completion, the reaction mixture was diluted with THF (20 mL), then successively was added H₂O (0.7 mL), aq. NaOH (0.7 mL, 15%), H₂O (2.1 mL) and dried over anhydrous sodium sulfate at 0 °C under N2. The reaction mixture was filtered, and the filtrate was concentrated in vacuum to give residue. The residue was purified by flash silica gel chromatography (ISCO®; 80 g SepaFlash® Silica Flash Column, Eluent of 0 - 10% MeOH / DCM gradient at 100 mL / min) to give compound benzyl N-[8-hydroxy-1-(7- hydroxyheptyl)octyl]-N-(1-methyl-4-piperidyl)carbamate (6h) (2.2 g, 4.35 mmol, 47.5% yield, 97.0% purity) as a colorless oil. ¹H NMR (400 MHz, CDCl₃) δ = 7.46 - 7.21 (m, 5H), 5.13 (s, 2H), 4.33 - 3.97 (m, 1H), 3.61 (t, J = 6.4 Hz, 4H), 3.17 - 2.78 (m, 3H), 2.50 - 2.09 (m, 6H), 2.00 (s, 2H), 1.75 - 1.39 (m, 10H), 1.38 - 1.09 (m, 18H). [00308] Step 8: 8-(((benzyloxy)carbonyl)(1-methylpiperidin-4-yl)amino)pentadecane-1,15- diyl bis(4,4-bis(pentyloxy)butanoate) (6i):

[00309] To a solution of benzyl N-[8-hydroxy-1-(7-hydroxyheptyl)octyl]-N-(1-methyl-4- piperidyl)carbamate (6h) (2.93 g, 5.97 mmol, 1.0 eq), 4,4-dipentoxybutanoic acid (3.26 g, 12.54 mmol, 2.1 eq), TEA (3.02 g, 29.85 mmol, 4.16 mL, 5.0 eq) and DMAP (2.19 g, 17.91 mmol, 3.0 eq) in DCM (1 mL) was added EDCI (2.86 g, 14.93 mmol, 2.5 eq). The mixture was stirred at 20 °C for 24 hours. The reaction mixture was partitioned between DCM (60 mL) and H2O (60 mL), the organic phase was washed with 0.5 N HCl (50 mL), then washed with NaHCO₃ (60 mL). The organic phase was separated, washed with brine (70 mL), dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure to give a residue. The residue was purified by flash silica gel chromatography (ISCO®; 40 g SepaFlash® Silica Flash Column, Eluent of 0 - 10% EtOAc/petroleum ether gradient at 100 mL / min) to give compound [8- [benzyloxycarbonyl-(1-methyl-4-piperidyl)amino]-15-(4,4-dipentoxybutanoyloxy)pentadecyl] 4,4-dipentoxybutanoate (6i) (2.5 g, 2.56 mmol, 42.9% yield) as a yellow oil. ¹H NMR (400 MHz, CDCl₃) δ = 7.45 - 7.28 (m, 5H), 5.4 - 5.0 (m, 2H), 4.50 (t, J = 5.6 Hz, 2H), 4.04 (t, J = 6.8 Hz, 4H), 3.60 - 3.53 (m, 4H), 3.45 - 3.36 (m, 4H), 2.89 (d, J = 10.4 Hz, 2H), 2.38 (t, J = 7.6 Hz, 4H), 2.25 (s, 4H), 2.04 - 1.85 (m, 8H), 1.66 - 1.47 (m, 18H), 1.37 - 1.17 (m, 36H), 0.90 (t, J = 6.4 Hz, 12H). [00310] Step 9: 8-((1-methylpiperidin-4-yl)amino)pentadecane-1,15-diyl bis(4,4- bis(pentyloxy)butanoate) (6j):

[00311] To a solution of [8-[benzyloxycarbonyl-(1-methyl-4-piperidyl)amino]-15-(4,4- dipentoxybutanoyloxy)pentadecyl] 4,4-dipentoxybutanoate (6i) (2.50 g, 2.56 mmol, 1.0 eq) in THF (15 mL) and MeOH (15 mL) was added Pd/C (10%, 200 mg), Pd(OH)2/C (20%, 200 mg) under N2. The suspension was degassed and purged with H23 times. The mixture was stirred under H₂ (15 PSI) at 40 °C for 1.5 hours. The reaction was filtered and concentrated. The residue was purified by flash silica gel chromatography (ISCO®; 20 g SepaFlash® Silica Flash Column, Eluent of 0 - 10% MeOH / DCM gradient at 100 mL / min) to give compound [15-(4,4- dipentoxybutanoyloxy)-8-[(1-methyl-4-piperidyl)amino]pentadecyl] 4,4-dipentoxybutanoate (6j) (1.97 g, 2.11 mmol, 82.2% yield, 90.0% purity) as a colorless oil. ¹H NMR (400 MHz, CDCl₃) δ = 4.50 (t, J = 5.6 Hz, 2H), 4.06 (t, J = 6.8 Hz, 4H), 3.52 - 3.51 (m, 4H), 3.49 - 3.47 (m, 4H), 2.81 (d, J = 11.4 Hz, 2H), 2.59 - 2.45 (m, 2H), 2.38 (t, J = 7.6 Hz, 4H), 2.28 (s, 3H), 2.04 - 1.97 (m, 2H), 1.96 - 1.90 (m, 4H), 1.86 (d, J = 11.6 Hz, 2H), 1.64 - 1.53 (m, 12H), 1.37 - 1.23 (m, 40H), 0.90 (t, J = 6.9 Hz, 12H). [00312] Step 10: 8-(((heptylthio)carbonyl)(1-methylpiperidin-4-yl)amino)pentadecane-1,15- diyl bis(4,4-bis(pentyloxy)butanoate) (6)

[00313] To a solution of [15-(4,4-dipentoxybutanoyloxy)-8-[(1-methyl-4- piperidyl)amino]pentadecyl] 4,4-dipentoxybutanoate (6j) (1.77 g, 2.10 mmol, 1.0 eq) and TEA (638.68 mg, 6.31 mmol, 878.51 μL, 3.0 eq) dissolved in dry DCM (30 mL) was added triphosgene (374.60 mg, 1.26 mmol, 0.6 eq) at 0 °C under N2. The resulting solution was stirred at 20 °C for 2 hours (checked by TLC) and concentrated under reduced pressure and kept under N₂. To a solution of heptane-1-thiol (834.83 mg, 6.31 mmol, 989.13 μL, 3.0 eq) dissolved in dry THF (30 mL) was added NaOH (252.45 mg, 6.31 mmol, 3.0 eq) at 0 °C under N2. To this resulting solution the above carbamoyl chloride, dissolved in THF (20 mL) was added via syringe slowly under N₂ at 0 °C. The resulting solution was stirred at 25 ° C for 15 hours. The reaction mixture was quenched by NH₄Cl (30 mL) at 0 °C and then diluted with EtOAc (20 mL). The aqueous phase was extracted with EtOAc (20 mL × 2). The combined organic phase was washed with brine (30 mL), dried with anhydrous sodium sulfate, filtered, and concentrated in vacuum to give residue. The residue was purified by flash silica gel chromatography (ISCO®; 20 g SepaFlash® Silica Flash Column, Eluent of 0 - 10% EtOAc/petroleum ether gradient at 80 mL/min). then was further separated by positive purification (column: Welch Ultimate XB CN 150 × 25 × 10 um; mobile phase: [Hexane - EtOH (0.1% IPAm)]; gradient: 0% - 30% B over 14 min) to give compound [15-(4,4-dipentoxybutanoyloxy)-8-[heptylsulfanylcarbonyl-(1-methyl-4- piperidyl)amino]pentadecyl] 4,4-dipentoxybutanoate (6) (428 mg, 428.15 μmol, 24.2% yield, 99.99% purity) as a yellow oil. LCMS: [M+H]⁺: 999.8; ¹H NMR (400 MHz, CDCl₃-d) δ = 4.50 (t, J = 5.6 Hz, 2H), 4.05 (t, J = 6.8 Hz, 4H), 3.89 - 3.66 (m, 1H), 3.64 - 3.53 (m, 4H), 3.45 - 3.35 (m, 4H), 3.01 - 2.90 (m, 2H), 2.89 - 2.70 (m, 4H), 2.38 (t, J = 7.6 Hz, 4H), 2.32 - 2.22 (m, 3H), 2.08 - 2.00 (m, 1H), 1.96 - 1.91 (m, 4H), 1.86 - 1.66 (m, 8H), 1.63 - 1.52 (m, 14H), 1.38 - 1.24 (m, 40H), 0.97 - 0.85 (m, 15H). Example 7: Synthesis of bis(2-butyloctyl) 10-((3- (dimethylamino)propyl)(octylsulfinyl)amino)nonadecanedioate (7):

[00314] Step 1: S-octyl ethanethioate (7a):

[00315] To a stirred mixture of octane-1-thiol (4 g, 27.34 mmol, 4.74 mL, 1.0 eq) and acetic oxide (5.86 g, 57.42 mmol, 5.39 mL, 2.1 eq), dichloromanganese tetrahydrate (1.62 g, 8.20 mmol, 0.3 eq) was added. The mixture was stirred at 60 °C for 16 hours. The reaction mixture was quenched with saturated aq. NaHCO3 (60 mL) and extracted with EtOAc (30 mL × 3). The organic layer was dried over anhydrous sodium sulfate and concentrated under reduced pressure. The residue was purified by flash silica gel chromatography (ISCO®; 80 g SepaFlash® Silica Flash Column, Eluent of 0 - 10% EtOAc/petroleum ether gradient at 100 mL / min) to give compound S-octyl ethanethioate (7a) (3.6 g, 19.12 mmol, 69.9% yield) as a colorless oil.¹H NMR (400 MHz, CDCl₃) δ = 2.86 (t, J = 7.2 Hz, 2H), 2.32 (s, 3H), 1.62 - 1.50 (m, 2H), 1.40 - 1.32 (m, 2H), 1.31 - 1.21 (m, 8H), 0.88 (t, J = 6.8 Hz, 3H). [00316] Step 2: octane-1-sulfinic chloride (7b):

[00317] A mixture of S-octyl ethanethioate (7a) (2.78 g, 14.76 mmol, 1.0 eq) in DCM (35 mL) was degassed and purged with N23 times, cool down to -20 °C, then Ac2O (1.51 g, 14.76 mmol, 1.39 mL, 1.0 eq) and sulfuryl chloride (3.98 g, 29.52 mmol, 2.95 mL, 2.0 eq) in DCM (2 mL) was added slowly via syringe and then the mixture was stirred at -20 °C for 30 min under N₂. The reaction mixture was concentrated under reduced pressure to give compound octane-1- sulfinyl chloride (7b) (2.91 g, crude) as a light yellow oil. [00318] Step 3: bis(2-butyloctyl) 10-((3- (dimethylamino)propyl)(octylsulfinyl)amino)nonadecanedioate (7)

[00319] To a solution of bis(2-butyloctyl) 10-[3- (dimethylamino)propylamino]nonadecanedioate (1h) (1 g, 1.31 mmol, 1.0 eq) , DMAP (31.93 mg, 261.34 μmol, 0.2 eq) and TEA (661.13 mg, 6.53 mmol, 909.39 μL, 5.0 eq) in DCM (20 mL) was added octane-1-sulfinyl chloride (7b) (1.03 g, 5.23 mmol, 4.0 eq) in DCM (6 mL) at 0 °C under N2. The mixture was stirred at 25 °C for 1 hour. The reaction was concentrated. The residue was purified by prep-HPLC (Formic Acid condition; column: phenyl-Hexyl; mobile phase: [water (Formic Acid) - MeOH]; gradient: 65% - 95% B over 10 min) and flash silica gel chromatography (ISCO®; 40 g SepaFlash® Silica Flash Column, Eluent of 0 - 5% MeOH / DCM gradient at 40 mL / min) to give compound bis(2-butyloctyl) 10-[3- (dimethylamino)propyl-octylsulfinyl-amino]nonadecanedioate (7) (226 mg, 241.76 μmol, 28.0% yield, 99.01% purity) as a yellow oil. LCMS: [M+H]⁺: 765.7; ¹H NMR (400 MHz, CDCl3) δ = 3.97 (d, J = 6.0 Hz, 4H), 3.42 - 3.28 (m, 1H), 3.05 - 2.95 (m, 1H), 2.85 - 2.70 (m, 2H), 2.69 - 2.58 (m, 1H), 2.46 - 2.36 (m, 2H), 2.36 - 2.25 (m, 10H), 1.89 - 1.71 (m, 2H), 1.65 - 1.57 (m, 8H), 1.52 - 1.47 (m, 2H), 1.44 - 1.35 (m, 4H), 1.34 - 1.21 (m, 60H), 0.96 - 0.81 (m, 15H). Example 8: Synthesis of 8-(((heptylthio)carbonyl)(2-(1-methylpyrrolidin-2- yl)ethyl)amino)pentadecane-1,15-diyl bis(4,4-bis(pentyloxy)butanoate) (8)

[00320] Step 1: Synthesis of dimethyl 8-((2-(1-methylpyrrolidin-2- yl)ethyl)amino)pentadecanedioate (8a)

[00321] A mixture of 2-(1-methylpyrrolidin-2-yl)ethanamine (6e) (4 g, 31.20 mmol, 4.52 mL, 1.0 eq), NaBH(OAc)3 (9.92 g, 46.80 mmol, 1.5 eq), HOAc (2.81 g, 46.80 mmol, 2.68 mL, 1.5 eq) in DCM (60 mL) was degassed and purged with N₂3 times, dimethyl 8- oxopentadecanedioate (6e) (10.79 g, 34.32 mmol, 1.1 eq) in DCM (20 mL) was added and then the mixture was stirred at 20 °C for 10 hours under N2. The mixture pH was adjusted to 9-10 with 1 N NaOH (30 mL), the aqueous phase was extracted with EtOAc (35 mL × 3). The combined organic phase was washed with brine (40 mL), dried with anhydrous sodium sulfate, filtered, and concentrated in vacuum to give crude product. The residue was purified by flash silica gel chromatography (ISCO®; 80 g SepaFlash® Silica Flash Column, Eluent of 0 ~ 10% MeOH / DCM gradient @ 100 mL / min) to give compound dimethyl 8-[2-(1-methylpyrrolidin- 2-yl)ethylamino]pentadecanedioate (8a) (7.97 g, 18.67 mmol, 59.8% yield) as a yellow oil. ¹H NMR (400 MHz, CDCl3) δ = 3.71 - 3.59 (m, 6H), 3.10 - 2.96 (m, 1H), 2.66 - 2.51 (m, 2H), 2.48 - 2.40 (m, 1H), 2.35 - 2.26 (m, 7H), 2.17 - 2.06 (m, 2H), 1.97 - 1.87 (m, 1H), 1.85 - 1.66 (m, 3H), 1.65 - 1.55 (m, 4H), 1.54 - 1.41 (m, 2H), 1.39 - 1.23 (m, 16H). [00322] Step 2: Synthesis of dimethyl 8-(((benzyloxy)carbonyl)(2-(1-methylpyrrolidin-2- yl)ethyl)amino)pentadecanedioate (8b)

[00323] A mixture of dimethyl 8-[2-(1-methylpyrrolidin-2-yl)ethylamino]pentadecanedioate (8a) (7.96 g, 18.66 mmol, 1.0 eq), DIEA (2.89 g, 22.39 mmol, 3.90 mL, 1.2 eq) in DCM (130 mL) was degassed and purged with N23 times, then CbzOSu (5.58 g, 22.39 mmol, 1.2 eq) was added at 0 °C, and the reaction was stirred at 25 °C for 3 hours. The reaction mixture was concentrated under reduced pressure to give a residue. The residue was purified by flash silica gel chromatography (ISCO®; 80 g SepaFlash® Silica Flash Column, Eluent of 0 ~ 10% MeOH / DCM gradient @100 mL / min) to give compound dimethyl 8-[benzyloxycarbonyl-[2-(1- methylpyrrolidin-2-yl)ethyl]amino]pentadecanedioate (8b) (9.97 g, 17.78 mmol, 95.3% yield) as a yellow oil. ¹H NMR (400 MHz, CDCl3) δ = 7.42 - 7.23 (m, 5H), 5.23 - 5.01 (m, 2H), 4.06 - 3.77 (m, 1H), 3.65 (s, 6H), 3.24 - 3.01 (m, 2H), 2.60 - 2.53 (m, 5H), 2.39 - 2.14 (m, 7H), 2.10 - 1.90 (m, 2H), 1.84 - 1.69 (m, 2H), 1.59 - 1.50 (m, 4H), 1.49 - 1.35 (m, 4H), 1.31 - 1.15 (m, 12H). [00324] Step 3: Synthesis of benzyl (1,15-dihydroxypentadecan-8-yl)(2-(1-methylpyrrolidin- 2-yl)ethyl)carbamate (8c):

[00325] A mixture of dimethyl 8-[benzyloxycarbonyl-[2-(1-methylpyrrolidin-2- yl)ethyl]amino]pentadecanedioate (8b) (9.97 g, 17.78 mmol, 1.0 eq) in THF (100 mL) was degassed and purged with N23 times and cooled down to 0 °C, then LiAlH4 (2.5 M, 14.93 mL, 2.1 eq) was added via syringe at 0 °C, and then the mixture was stirred at 25 °C for 1.5 hours under N2. After completion, the reaction mixture was diluted with THF (50 mL), then successively was added H2O (1.42 mL), aq. NaOH (1.42 mL, 15%), H2O (4.26 mL) and dried over anhydrous sodium sulfate at 0 °C under N₂. The reaction mixture was filtered and the filtrate was concentrated in vacuum to give residue. The residue was purified by flash silica gel chromatography (ISCO®; 80 g SepaFlash® Silica Flash Column, Eluent of 0 ~ 10% MeOH / DCM gradient @ 100 mL / min) to give compound benzyl N-[8-hydroxy-1-(7- hydroxyheptyl)octyl]-N-[2-(1-methylpyrrolidin-2-yl)ethyl]carbamate (8c) (6.53 g, 12.94 mmol, 73.4% yield) as a colorless oil. ¹H NMR (400 MHz, CDCl3) δ = 7.44 - 7.26 (m, 5H), 5.26 - 5.05 (m, 2H), 4.14 - 3.96 (m, 1H), 3.68 -3.54 (m, 4H), 3.12 - 2.92 (m, 3H), 2.24 - 2.17 (m, 2H), 2.12 - 2.04 (m, 2H), 1.97 - 1.85 (m, 2H), 1.84 - 1.84 (m, 1H), 1.79 - 1.62 (m, 2H), 1.57 - 1.52 (m, 4H), 1.49 - 1.40 (m, 4H), 1.39 - 1.14 (m, 18H). [00326] Step 4: Synthesis of 8-(((benzyloxy)carbonyl)(2-(1-methylpyrrolidin-2- yl)ethyl)amino)pentadecane-1,15-diyl bis(4,4-bis(pentyloxy)butanoate) (8d)

[00327] To a solution of benzyl N-[8-hydroxy-1-(7-hydroxyheptyl)octyl]-N-[2-(1- methylpyrrolidin-2-yl)ethyl]carbamate (8c) (2 g, 3.96 mmol, 1.0 eq), 4,4-dipentoxybutanoic acid (2.58 g, 9.91 mmol, 2.5 eq), TEA (2.00 g, 19.81 mmol, 2.76 mL, 5.0 eq) and DMAP (1.45 g, 11.89 mmol, 3.0 eq) in DCM (50 mL) was added EDCI (1.90 g, 9.91 mmol, 2.5 eq). The mixture was stirred at 35 °C for 24 hours. The reaction mixture was partitioned between DCM (60 mL) and H₂O (60 mL). The organic phase was washed with 0.5 N HCl (50 mL), then washed with NaHCO3 (60 mL). The organic phase was separated, washed with brine (70 mL), dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure to give a residue. The residue was purified by flash silica gel chromatography (ISCO®; 40 g SepaFlash® Silica Flash Column, Eluent of 0 ~ 10% MeOH / DCM gradient @ 100 mL / min) to give compound [8-[benzyloxycarbonyl-[2-(1-methylpyrrolidin-2-yl)ethyl]amino]-15-(4,4- dipentoxybutanoyloxy)pentadecyl] 4,4-dipentoxybutanoate (8d) (3.16 g, 3.16 mmol, 79.8% yield, 99.0% purity) as a yellow oil.¹H NMR (400 MHz, CDCl₃) δ = 7.39 - 7.24 (m, 5H), 5.22 - 5.04 (m, 2H), 4.57 - 4.41 (m, 2H), 4.15 - 3.98 (m, 5H), 3.64 - 3.55 (m, 4H), 3.44 - 3.36 (m, 4H), 3.24 - 3.00 (m, 3H), 2.43 - 2.34 (m, 5H), 2.24 - 2.15 (m, 2H), 2.12 - 1.98 (m, 2H), 1.97 - 1.87 (m, 6H), 1.61 - 1.53 (m, 14H), 1.47 - 1.39 (m, 4H), 1.37 - 1.24 (m, 34H), 0.91 - 0.88 (m, 12H). [00328] Step 5: Synthesis of 8-((2-(1-methylpyrrolidin-2-yl)ethyl)amino)pentadecane-1,15- diyl bis(4,4-bis(pentyloxy)butanoate) (8e)

[00329] To a solution of [8-[benzyloxycarbonyl-[2-(1-methylpyrrolidin-2-yl)ethyl]amino]-15- (4,4-dipentoxybutanoyloxy)pentadecyl] 4,4-dipentoxybutanoate (8d) (2.09 g, 2.11 mmol, 1.0 eq) in THF (20 mL) and MeOH (20 mL) was added Pd / C (10%, 300 mg), Pd(OH)2 / C (10%, 300 mg) under N2. The suspension was degassed and purged with H23 times. The mixture was stirred under H₂ (15 PSI) at 40 °C for 16 hours. The reaction was filtered and concentrated. The residue was purified by flash silica gel chromatography (ISCO®; 20 g SepaFlash® Silica Flash Column, Eluent of 0 ~ 15% MeOH / DCM gradient @ 100 mL / min) to give compound [15- (4,4-dipentoxybutanoyloxy)-8-[2-(1-methylpyrrolidin-2-yl)ethylamino]pentadecyl] 4,4- dipentoxybutanoate (8e) (1.52 g, 1.78 mmol, 84.1% yield) as a yellow oil. ¹H NMR (400 MHz, CDCl3) δ = 4.50 (t, J = 5.2 Hz, 2H), 4.06 (t, J = 6.8 Hz, 4H), 3.62 - 3.50 (m, 4H), 3.46 - 3.35 (m, 4H), 3.11 - 3.00 (m, 1H), 2.70 - 2.56 (m, 2H), 2.50 - 2.44 (m, 1H), 2.38 (t, J = 7.2 Hz, 4H), 2.35 - 2.29 (m, 3H), 2.17 - 2.10 (m, 2H), 1.98 - 1.88 (m, 5H), 1.86 - 1.67 (m, 3H), 1.66 - 1.50 (m, 14H), 1.42 - 1.24 (m, 36H), 0.90 (t, J = 6.4 Hz, 12H). [00330] Step 6: Synthesis of 8-(((heptylthio)carbonyl)(2-(1-methylpyrrolidin-2- yl)ethyl)amino)pentadecane-1,15-diyl bis(4,4-bis(pentyloxy)butanoate) (8)

[00331] To a solution of [15-(4,4-dipentoxybutanoyloxy)-8-[2-(1-methylpyrrolidin-2- yl)ethylamino]pentadecyl] 4,4-dipentoxybutanoate (8e) (1.52 g, 1.78 mmol, 1.0 eq) and TEA (539.47 mg, 5.33 mmol, 742.06 μL, 3.0 eq) dissolved in dry DCM (30 mL) was added triphosgene (316.41 mg, 1.07 mmol, 0.6 eq) at 0 °C under N₂. The resulting solution was stirred at 25 °C for 2 hours (checked by TLC) and concentrated under reduced pressure and kept under N2. To a solution of heptane-1-thiol (691.76 mg, 5.23 mmol, 819.62 μL, 3.0 eq) dissolved in dry THF (30 mL) was added NaOH (209.19 mg, 5.23 mmol, 3.0 eq) at 0 °C under N2. To this resulting solution the above carbamoyl chloride, dissolved in THF (20 mL) was added via syringe slowly under N2 at 0 °C. The resulting solution was stirred at 25 ° C for 15 hours. The reaction mixture was quenched by NH4Cl (10 mL) at 0 °C and then diluted with DCM (5 mL). The aqueous phase was extracted with EtOAc (20 mL × 2). The combined organic phase was washed with brine (30 mL), dried with anhydrous sodium sulfate, filtered, and concentrated under vacuum to give residue. The residue was purified by flash silica gel chromatography (ISCO®; 20 g SepaFlash® Silica Flash Column, Eluent of 0 ~ 5% MeOH / DCM gradient @ 50 mL / min) to give compound [15-(4,4-dipentoxybutanoyloxy)-8-[heptylsulfanylcarbonyl-[2-(1- methylpyrrolidin-2-yl)ethyl]amino]pentadecyl] 4,4-dipentoxybutanoate (8, 462 mg, 450.11 μmol, 30.4% yield, 98.75% purity) as a yellow oil. LCMS: [M+H]⁺: 1013.8; ¹H NMR (400 MHz, CDCl₃) δ = 4.49 (t, J = 5.6 Hz, 2H), 4.04 (t, J = 6.8 Hz, 4H), 3.90 - 3.69 (m, 1H), 3.62 - 3.53 (m, 4H), 3.44 - 3.36 (m, 4H), 3.29 - 3.16 (m, 1H), 3.15 - 3.03 (m, 2H), 2.95 - 2.83 (m, 2H), 2.43 - 2.31 (m, 7H), 2.22 - 1.98 (m, 4H), 1.96 - 1.90 (m, 4H), 1.87 - 1.65 (m, 3H), 1.61 - 1.52 (m, 14H), 1.48 - 1.41 (m, 4H), 1.39 - 1.20 (m, 42H), 0.97 - 0.82 (m, 15H). Example 9: Synthesis of 8-(((heptylthio)carbonyl)((1-methylpiperidin-4- yl)methyl)amino)pentadecane-1,15-diyl bis(4,4-bis(pentyloxy)butanoate) (9) [00332] Step 1: 4,4-bis(pentyloxy)butanenitrile (9a):

[00333] A mixture of 4,4-dimethoxybutanenitrile (15 g, 116.14 mmol, 1.0 eq) and PPTS (1.46 g, 5.81 mmol, 0.1 eq) inpentan-1-ol (30.71 g, 348.41 mmol, 37.87 mL, 3.0 eq) was degassed and purged with N₂3 times, and then the mixture was stirred at 110 °C for 16 h under N₂. The reaction was concentrated. The residue was purified by flash silica gel chromatography (ISCO®; 120 g SepaFlash® Silica Flash Column, Eluent of 0 ~ 3% EtOAc / PE gradient @ 100 mL / min) to give 4,4-dipentoxybutanenitrile (9a) (15.88 g, 59.21 mmol, 51% yield, 90% purity) as a yellow oil.¹H NMR (400 MHz, CDCl₃) δ = 4.56 (t, J = 5.2 Hz, 1H), 3.65 - 3.55 (m, 2H), 3.49 - 3.37 (m, 2H), 2.42 (t, J = 7.4 Hz, 2H), 1.96 - 1.89 (m, 2H), 1.63 - 1.54 (m, 4H), 1.40 - 1.28 (m, 8H), 0.97 - 0.85 (m, 6H). [00334] Step 2: 4,4-bis(pentyloxy)butanoic acid (9b):

[00335] A mixture of 4,4-dipentoxybutanenitrile (9a) (15.88 g, 65.79 mmol, 1.0 eq) and KOH (11.07 g, 197.37 mmol, 3.0 eq) in EtOH (80 mL) and H₂O (80 mL) was degassed and purged with N23 times, and then the mixture was stirred at 110 °C for 5 h under N2. The reaction mixture cooled down to room temperature and concentrated ,the diluted with EtOAc (200 mL), pH adjust to pH = 5 by addition 1 N HCl, and extracted with EtOAc 200 mL (100 mL × 2). The combined organic layers were washed with brine (200 mL) , dried over anhydrous sodium sulfate, filtered and concentrated under reduced pressure to give a residue. The residue was purified by flash silica gel chromatography (ISCO®; 80 g SepaFlash® Silica Flash Column, Eluent of 0 ~ 30% EtOAc / PE gradient @ 100 mL / min) to give 4,4-dipentoxybutanoic acid (9b) (11 g, 35.91 mmol, 54.6% yield, 85% purity) as a yellow oil. ¹H NMR (400 MHz, CDCl₃) δ = 4.45 (t, J = 5.6 Hz, 1H), 3.55 - 3.46 (m, 2H), 3.39 - 3.29 (m, 2H), 2.43 - 2.31 (m, 2H), 1.92 - 1.81 (m, 2H), 1.55 - 1.45 (m, 4H), 1.30 - 1.20 (m, 8H), 0.83 (t, J = 6.8 Hz, 6H). [00336] Step 3: dimethyl 8-isocyano-8-tosylpentadecanedioate (9c)

[00337] To a 2000 mL three neck round bottom flask under N₂, was added DMSO (1000 mL), followed by the addition of NaH (19.98 g, 499.39 mmol, 60% purity, 2.5 eq) in portions over a period of 0.5 h. The mixture was allowed to stir for 0.5 h at 20 °C following the addition, then 1- (isocyanomethylsulfonyl)-4-methyl-benzene (39 g, 199.76 mmol, 1.0 eq) was added in portions over 0.5 h, followed by the addition of TBAI (7.38 g, 19.98 mmol, 0.1 eq) in one portion. The resulting mixture was stirred for 0.5 h at 20 °C, then methyl 7-bromoheptanoate (93.59 g, 419.49 mmol, 2.1 eq) was added over a period of 0.5 hour. The mixture was stirred at 20 °C for 1 h. The reaction mixture was quenched by addition water (300 mL) at 0 °C, and then extracted with PE / EtOAc = 10 / 1 (300 mL × 2). The combined organic layers were washed with brine (500 mL), dried over anhydrous sodium sulfate, filtered and concentrated under reduced pressure to give dimethyl 8-isocyano-8-(p-tolylsulfonyl)pentadecanedioate (9c) (90 g, crude) as a black oil. [00338] Step 4: dimethyl 8-oxopentadecanedioate (9d)

[00339] To a 1000 mL three neck round bottom flask was added DCM (500 mL) , followed by the addition of dimethyl 8-isocyano-8-(p-tolylsulfonyl)pentadecanedioate (9c) (90 g, 187.65 mmol, 1.0 eq), under nitrogen, HCl (12 M, 112 mL, 7.2 eq) was added in portions over a period of 1 h. The two-phase mixture was stirred for 12 h at 25 °C. The reaction mixture was quenched by addition water (150 mL) at 0 °C, and then extracted with PE (150 mL × 2). The combined organic layers were washed with brine (200 mL), dried over anhydrous sodium sulfate, filtered and concentrated under reduced pressure to give a residue. The residue was purified by column chromatography (SiO₂, PE / EtOAc = 1 / 0 to 10 / 1) to give dimethyl 8-oxopentadecanedioate (9d) (37.8 g, 108.20 mmol, 68% yield, 90% purity) as a colorless oil. ¹H NMR (400 MHz, CDCl3) δ = 3.73 - 3.59 (m, 6H), 2.37 (t, J = 7.2 Hz, 4H), 2.33 - 2.24 (m, 4H), 1.66 - 1.52 (m, 8H), 1.34 - 1.24 (m, 8H). [00340] Step 5: dimethyl 8-(((1-methylpiperidin-4-yl)methyl)amino)pentadecanedioate (9e):

[00341] A mixture of (1-methyl-4-piperidyl)methanamine (3.5 g, 27.30 mmol, 1.0 eq), NaBH(OAc)3 (8.68 g, 40.95 mmol, 1.5 eq), HOAc (2.46 g, 40.95 mmol, 2.34 mL, 1.5 eq) in DCM (70 mL) was degassed and purged with N23 times, dimethyl 8-oxopentadecanedioate (9d) (9.44 g, 30.03 mmol, 1.1 eq) in DCM (20 mL) was added and then the mixture was stirred at 20 °C for 10 h under N₂. The mixture pH was adjusted to 9 - 10 was with 1 M NaOH (20 mL), the aqueous phase was extracted with EtOAc (35 mL × 3). The combined organic phase was washed with brine (40 mL), dried with anhydrous sodium sulfate, filtered and concentrated in vacuum to give crude product. The residue was purified by flash silica gel chromatography (ISCO®; 80 g SepaFlash® Silica Flash Column, Eluent of 0~10% MeOH / DCM gradient @ 100 mL / min) to give dimethyl 8-[(1-methyl-4-piperidyl)methylamino]pentadecanedioate (9e) (11.2 g, 25.73 mmol, 94.3% yield, 98% purity) as a yellow oil. ¹H NMR (400 MHz, CDCl₃) δ = 3.64 (s, 6H), 2.83 (d, J = 11.2 Hz, 2H), 2.46 - 2.34 (m, 3H), 2.28 (t, J = 7.6 Hz, 4H), 2.24 (s, 3H), 1.90 (t, J = 10.8 Hz, 2H), 1.70 (d, J = 12.4 Hz, 2H), 1.65 - 1.49 (m, 5H), 1.33 - 1.21 (m, 18H). [00342] Step 6: dimethyl 8-(((benzyloxy)carbonyl)((1-methylpiperidin-4- yl)methyl)amino)pentadecanedioate (9f)

[00343] A mixture of dimethyl 8-[(1-methyl-4-piperidyl)methylamino]pentadecanedioate (9e) (10.98 g, 25.74 mmol, 1.0 eq), DIEA (6.65 g, 51.47 mmol, 8.96 mL, 2.0 eq) in DCM (180 mL) was degassed and purged with N₂3 times, then benzyl (2,5-dioxopyrrolidin-1-yl) carbonate (7.69 g, 30.88 mmol, 1.2 eq) was added at 0 °C, and the reaction was stirred at 25 °C for 3 h. The reaction mixture was concentrated under reduced pressure to give a residue. The residue was purified by flash silica gel chromatography (ISCO®; 80 g SepaFlash® Silica Flash Column, Eluent of 0 ~ 5% MeOH / DCM gradient @ 100 mL / min) to give dimethyl 8- [benzyloxycarbonyl-[(1-methyl-4-piperidyl)methyl]amino]pentadecanedioate (9f) (9.8 g, 16.60 mmol, 64.5% yield, 95% purity) as a yellow oil. ¹H NMR (400 MHz, DMSO-d₆) δ = 7.41 - 7.18 (m, 5H), 5.04 (s, 2H), 3.57 (s, 6H), 2.95 (d, J = 6.0 Hz, 2H), 2.80 - 2.66 (m, 2H), 2.29 - 2.20 (m, 4H), 2.19 - 2.06 (m, 3H), 1.86 - 1.64 (m, 2H), 1.58 - 1.33 (m, 11H), 1.29 - 1.03 (m, 15H). [00344] Step 7: benzyl (1,15-dihydroxypentadecan-8-yl)((1-methylpiperidin-4- yl)methyl)carbamate (9g):

[00345] A mixture of dimethyl 8-[benzyloxycarbonyl-[(1-methyl-4- piperidyl)methyl]amino]pentadecanedioate (9f) (9.3 g, 16.58 mmol, 1.0 eq) in THF (100 mL) was degassed and purged with N23 times and cooled down to 0 °C, then LiAlH4 (2.5 M, 13.93 mL, 2.1 eq) was added via injection at 0 °C, and then the mixture was stirred at 20 °C for 1.5 h under N2. After completion, the reaction mixture was diluted with THF (2 mL), then successively was added H2O (1.32 mL), aq. NaOH (1.32 mL, 15%), H2O (3.64 mL) and dried over anhydrous sodium sulfate at 0 °C under N₂. The reaction mixture was filtered and the filtrate was concentrated in vacuum to give residue. The residue was purified by flash silica gel chromatography (ISCO®; 80 g SepaFlash® Silica Flash Column, Eluent of 0 ~ 10% MeOH / DCM gradient @ 100 mL / min) to give benzyl N-[8-hydroxy-1-(7-hydroxyheptyl)octyl]-N-[(1- methyl-4-piperidyl)methyl]carbamate (9g) (7.17 g, 12.78 mmol, 51.4% yield, 90% purity) as a colorless oil. ¹H NMR (400 MHz, CDCl3) δ =7.40 - 7.25 (m, 5H), 5.10 (s, 2H), 3.86 - 3.68 (m, 1H), 3.61 (t, J = 6.4 Hz, 4H), 3.46 (s, 1H), 3.05 - 2.94 (m, 2H), 2.92 - 2.77 (m, 2H), 2.32 - 2.22 (m, 3H), 2.04 - 1.90 (m, 2H), 1.89 - 1.76 (m, 2H), 1.75 - 1.65 (m, 2H), 1.64 - 1.46 (m, 8H), 1.35 - 1.16 (m, 18H). [00346] Step 8: 8-(((benzyloxy)carbonyl)((1-methylpiperidin-4-yl)methyl)amino)pentadecane- 1,15-diyl bis(4,4-bis(pentyloxy)butanoate) (9h)

[00347] To a solution of benzyl N-[8-hydroxy-1-(7-hydroxyheptyl)octyl]-N-[(1-methyl-4- piperidyl)methyl]carbamate (9g) (2 g, 3.96 mmol, 1.0 eq), 4,4-dipentoxybutanoic acid (9b) (2.58 g, 9.91 mmol, 2.5 eq), TEA (2.00 g, 19.81 mmol, 2.76 mL, 5.0 eq) and DMAP (1.45 g, 11.89 mmol, 3.0 eq) in DCM (50 mL) was added EDCI (1.90 g, 9.91 mmol, 2.5 eq). The mixture was stirred at 35 °C for 24hr. The reaction mixture was partitioned between DCM (10 mL) and H2O (20 mL). The organic phase was separated, washed with brine 20 mL (10 mL × 2), dried over anhydrous sodium sulfate, filtered and concentrated under reduced pressure to give a residue. The residue was purified by flash silica gel chromatography (ISCO®; 40 g SepaFlash® Silica Flash Column, Eluent of 0 ~ 5% MeOH / DCM gradient @ 60 mL / min) to give [8- [benzyloxycarbonyl-[(1-methyl-4-piperidyl)methyl]amino]-15-(4,4- dipentoxybutanoyloxy)pentadecyl] 4,4-dipentoxybutanoate (9h) (1.5 g, 1.44 mmol, 36.4% yield, 95% purity) as a yellow oil. ¹H NMR (400 MHz, CDCl3) δ = 7.39 - 7.25 (m, 5H), 4.49 (t, J = 5.6 Hz, 2H), 4.04 (t, J = 6.8 Hz, 4H), 3.63 - 3.50 (m, 4H), 3.47 - 3.32 (m, 4H), 3.06 - 2.95 (m, 4H), 2.94 - 2.81 (m, 2H), 2.38 (t, J = 7.6 Hz, 4H), 2.28 (d, J = 16.4 Hz, 3H), 2.03 - 1.89 (m, 5H), 1.83 (s, 1H), 1.70 (d, J = 8.8 Hz, 2H), 1.64 - 1.52 (m, 14H), 1.48 (d, J = 6.0 Hz, 2H), 1.40 - 1.14 (m, 36H), 1.00 - 0.83 (m, 12H). [00348] Step 9: 8-(((1-methylpiperidin-4-yl)methyl)amino)pentadecane-1,15-diyl bis(4,4- bis(pentyloxy)butanoate) (9i)

[00349] To a solution of [8-[benzyloxycarbonyl-[(1-methyl-4-piperidyl)methyl]amino]-15- (4,4-dipentoxybutanoyloxy)pentadecyl] 4,4-dipentoxybutanoate (9h) (2.3 g, 2.32 mmol, 1.0 eq) in MeOH (10 mL) and THF (10 mL) were added Pd/C (10%, 100 mg) and Pd(OH)2/C (10%, 100 mg) under N2. The suspension was degassed and purged with H23 times. The mixture was stirred under H₂ (15 PSI ) at 40 °C for 2 h. The reaction was filtered and concentrated. The residue was purified by flash silica gel chromatography (ISCO®; 20 g SepaFlash® Silica Flash Column, Eluent of 0 ~ 15% MeOH / DCM gradient @ 100 mL / min) to give [15-(4,4- dipentoxybutanoyloxy)-8-[(1-methyl-4-piperidyl)methylamino]pentadecyl] 4,4- dipentoxybutanoate (9i) (1.5 g, 1.58 mmol, 69.4% yield, 90% purity) as a yellow oil. ¹H NMR (400 MHz, CDCl3) δ = 4.50 (t, J = 5.6 Hz, 2H), 4.06 (t, J = 6.8 Hz, 4H), 3.65 - 3.50 (m, 4H), 3.46 - 3.32 (m, 4H), 3.01 (s, 1H), 2.95 (d, J = 11.2 Hz, 2H), 2.46 (d, J = 6.0 Hz, 2H), 2.43 - 2.29 (m, 8H), 2.10 - 1.99 (m, 2H), 1.98 - 1.90 (m, 4H), 1.78 (d, J = 10.4 Hz, 2H), 1.64 - 1.50 (m, 12H), 1.40 - 1.23 (m, 38H), 0.94 - 0.82 (m, 12H). [00350] Step 10: 8-((chlorocarbonyl)((1-methylpiperidin-4-yl)methyl)amino)pentadecane- 1,15-diyl bis(4,4-bis(pentyloxy)butanoate) (9j)

[00351] To a solution of [15-(4,4-dipentoxybutanoyloxy)-8-[(1-methyl-4- piperidyl)methylamino]pentadecyl] 4,4-dipentoxybutanoate (9i) (1.5 g, 1.75 mmol, 1.0 eq) and TEA (532.38 mg, 5.26 mmol, 732.29 μL, 3.0 eq) dissolved in dry DCM (20 mL) was added triphosgene (312.25 mg, 1.05 mmol, 0.6 eq) at 0 °C under N₂. The resulting solution was stirred at 25 °C for 2 h and concentrated under reduced pressure to give [8-[chlorocarbonyl-[(1-methyl- 4-piperidyl)methyl]amino]-15-(4,4-dipentoxybutanoyloxy)pentadecyl] 4,4-dipentoxybutanoate (9j) (1.61 g, crude) as a yellow solid. [00352] Step 11: 8-(((heptylthio)carbonyl)((1-methylpiperidin-4- yl)methyl)amino)pentadecane-1,15-diyl bis(4,4-bis(pentyloxy)butanoate) (9)

[00353] To a solution of heptane-1-thiol (696.08 mg, 5.26 mmol, 824.74 μL, 3.0 eq) dissolved in dry THF (20 mL) was added NaOH (210.49 mg, 5.26 mmol, 3.0 eq) at 0 °C under N₂. Then [8-[chlorocarbonyl-[(1-methyl-4-piperidyl)methyl]amino]-15-(4,4- dipentoxybutanoyloxy)pentadecyl] 4,4-dipentoxybutanoate (9j) (1.61 g, 1.75 mmol, 1.0 eq) dissolved in THF (10 mL) was added via syringe slowly under N₂ at 0 °C. The resulting solution was stirred at 25 ° C for 4 h. The reaction mixture was quenched by NH4Cl (10 mL) at 0 °C and then diluted with EtOAc (5 mL). The aqueous phase was extracted with EtOAc (20 mL × 2). The combined organic phase was washed with brine (30 mL), dried with anhydrous sodium sulfate, filtered and concentrated in vacuum to give residue. The residue was purified by flash silica gel chromatography (ISCO®; 20 g SepaFlash® Silica Flash Column, Eluent of 0~6% MeOH / DCM gradient @ 50 mL / min) to give [15-(4,4-dipentoxybutanoyloxy)-8- [heptylsulfanylcarbonyl-[(1-methyl-4-piperidyl)methyl]amino]pentadecyl] 4,4- dipentoxybutanoate (9) (950 mg, 937.18 μmol, 63.3% yield, 99.99% purity) as a yellow oil. LCMS: [M+H]⁺: 1013.9¹H NMR (400 MHz, CDCl₃) δ = 4.50 (t, J = 5.6 Hz, 2H), 4.05 (t, J = 6.8 Hz, 4H), 3.81 (s, 1H), 3.64 -3.54 (m, 4H), 3.47 - 3.37 (m, 4H), 3.19 - 2.92 (m, 4H), 2.87 (t, J = 7.2 Hz, 2H), 2.42 - 2.31 (m, 7H), 2.17 - 1.99 (m, 2H), 1.96 - 1.90 (m, 4H), 1.89 - 1.79 (m, 1H), 1.77 - 1.70 (m, 2H), 1.65 - 1.54 (m, 14H), 1.47 - 1.40 (m, 4H), 1.40 - 1.20 (m, 42H), 0.98 - 0.82 (m, 15H). Example 10: Synthesis of 8-(((heptylthio)carbonyl)((1-methylpiperidin-4- yl)methyl)amino)pentadecane-1,15-diyl bis(4,4-bis(heptyloxy)butanoate) (10) [00354] Step 1: 4,4-bis(heptyloxy)butanenitrile (10a)

_, 10a [00355] A mixture of 4,4-dimethoxybutanenitrile (10 g, 77.43 mmol, 1.0 eq) and PPTS (972.86 mg, 3.87 mmol, 0.05 eq) inheptan-1-ol (26.99 g, 232.28 mmol, 32.84 mL, 3.0 eq) was degassed and purged with N₂3 times, and then the mixture was stirred at 110 °C for 16 h under N2. The reaction was concentrated. The residue was purified by flash silica gel chromatography (ISCO®; 120 g SepaFlash® Silica Flash Column, Eluent of 0 ~ 5% EtOAc / PE gradient @ 100 mL / min) to give 4,4-diheptoxybutanenitrile (10a) (7.7 g, 25.88 mmol, 33.4% yield) as a yellow oil. ¹H NMR (400 MHz, CDCl₃) δ = 4.55 (t, J = 5.2 Hz, 1H), 3.63 - 3.56 (m, 2H), 3.48 - 3.39 (m, 2H), 2.42 (t, J = 7.2 Hz, 2H), 1.98 - 1.90 (m, 2H), 1.61 - 1.54 (m, 4H), 1.36 - 1.25 (m, 16H), 0.93 - 0.85 (m, 6H). [00356] Step 2: 4,4-bis(heptyloxy)butanoic acid (10b)

[00357] A mixture of 4,4-diheptoxybutanenitrile (10a) (7.7 g, 25.88 mmol, 1.0 eq) and KOH (4.36 g, 77.65 mmol, 3.0 eq) in EtOH (30 mL) and H2O (30 mL) was degassed and purged with N₂3 times, and then the mixture was stirred at 110 °C for 3 h under N₂. The reaction mixture cooled down to room temperature, diluted with EtOAc 30 mL, pH adjust to pH = 5 by addition 1N HCl, and extracted with EtOAc 20 mL (10 mL × 2). The combined organic layers were washed with brine (30 mL), dried over anhydrous sodium sulfate, filtered and concentrated under reduced pressure to give a residue. The residue was purified by flash silica gel chromatography (ISCO®; 80 g SepaFlash® Silica Flash Column, Eluent of 0 ~ 30% EtOAc / PE gradient @ 100 mL / min) to give 4,4-diheptoxybutanoic acid (10b) (8.6 g, 24.46 mmol, 63% yield, 90% purity) as a yellow oil. ¹H NMR (400 MHz, CDCl₃) δ = 4.52 (t, J = 5.2 Hz, 1H), 3.62 - 3.53 (m, 2H), 3.49 - 3.34 (m, 2H), 2.51 - 2.40 (m, 2H), 2.03 - 1.88 (m, 2H), 1.61 - 1.52 (m, 4H), 1.34 - 1.25 (m, 16H), 0.89 (t, J = 6.8 Hz, 6H). [00358] Step 3: 8-(((benzyloxy)carbonyl)((1-methylpiperidin-4-yl)methyl)amino)pentadecane- 1,15-diyl bis(4,4-bis(heptyloxy)butanoate) (10c)

1_0c [00359] To a solution of benzyl N-[8-hydroxy-1-(7-hydroxyheptyl)octyl]-N-[(1-methyl-4- piperidyl)methyl]carbamate (9g) (3.5 g, 6.93 mmol, 1.0 eq), 4,4-diheptoxybutanoic acid (10b) (6.58 g, 20.80 mmol, 3.0 eq), TEA (3.51 g, 34.67 mmol, 4.83 mL, 5.0 eq) and DMAP (2.54 g, 20.80 mmol, 3.0 eq) in DCM (40 mL) was added EDCI (3.32 g, 17.34 mmol, 2.5 eq) . The mixture was stirred at 25 °C for 16 h. The reaction mixture was partitioned between DCM (30 mL) and H2O (20 mL). The organic phase was separated, washed with brine 20 mL (10 mL × 2), dried over anhydrous sodium sulfate, filtered and concentrated under reduced pressure to give a residue. The residue was purified by flash silica gel chromatography (ISCO®; 80 g SepaFlash® Silica Flash Column, Eluent of 0 ~ 10% EtOAc / PE gradient @ 80 mL / min) to give [8- [benzyloxycarbonyl-[(1-methyl-4-piperidyl)methyl]amino]-15-(4,4- diheptoxybutanoyloxy)pentadecyl] 4,4-diheptoxybutanoate (10c) (2.2 g, 1.60 mmol, 23% yield, 80% purity) as a yellow oil. ¹H NMR (400 MHz, CDCl3) δ = 7.40 - 7.27 (m, 5H), 5.11 (s, 2H), 4.49 (t, J = 5.6 Hz, 2H), 4.04 (t, J = 6.8 Hz, 4H), 3.63 - 3.53 (m, 4H), 3.45 - 3.35 (m, 4H), 3.08 - 2.96 (m, 3H), 2.93 - 2.83 (m, 1H), 2.47 - 2.32 (m, 6H), 2.30 (s, 1H), 2.14 (s, 1H), 1.99 - 1.87 (m, 5H), 1.86 - 1.69 (m, 2H), 1.64 - 1.52 (m, 14H), 1.50 - 1.41 (m, 4H), 1.37 - 1.15 (m, 50H), 0.97 - 0.80 (m, 12H). [00360] Step 4: 8-(((1-methylpiperidin-4-yl)methyl)amino)pentadecane-1,15-diyl bis(4,4- bis(heptyloxy)butanoate) (10d)

[00361] To a solution of [8-[benzyloxycarbonyl-[(1-methyl-4-piperidyl)methyl]amino]-15- (4,4-diheptoxybutanoyloxy)pentadecyl] 4,4-diheptoxybutanoate (10c) (2.2 g, 2.00 mmol, 1.0 eq) in THF (10 mL) and MeOH (10 mL) was added Pd/C (10%, 100 mg) Pd(OH)₂/C (10%, 100 mg) under N₂. The suspension was degassed and purged with H₂3 times. The mixture was stirred under H2 (15 PSI ) at 40 °C for 2 h. The reaction was filtered and concentrated. The residue was purified by flash silica gel chromatography (ISCO®; 20 g SepaFlash® Silica Flash Column, Eluent of 0 ~ 15% EtOAc / PE gradient @ 100 mL / min) to give [15-(4,4- diheptoxybutanoyloxy)-8-[(1-methyl-4-piperidyl)methylamino]pentadecyl] 4,4- diheptoxybutanoate (10d) (1.53 g, 1.42 mmol, 71.27% yield, 90% purity) as a yellow oil. ¹H NMR (400 MHz, CDCl₃) δ = 4.49 (t, J = 5.6 Hz, 2H), 4.05 (t, J = 6.8 Hz, 4H), 3.64 - 3.49 (m, 4H), 3.45 - 3.28 (m, 4H), 2.98 - 2.80 (m, 2H), 2.46 - 2.42 (m, 2H), 2.38 (t, J = 7.6 Hz, 4H), 2.32 - 2.26 (m, 3H), 2.02 - 1.89 (m, 6H), 1.80 - 1.70 (m, 2H), 1.62 - 1.51 (m, 11H), 1.47 - 1.17 (m, 58H), 0.95 - 0.79 (m, 12H). [00362] Step 5: 8-((chlorocarbonyl)((1-methylpiperidin-4-yl)methyl)amino)pentadecane-1,15- diyl bis(4,4-bis(heptyloxy)butanoate) (10e)

[00363] To a solution of [15-(4,4-diheptoxybutanoyloxy)-8-[(1-methyl-4- piperidyl)methylamino]pentadecyl] 4,4-diheptoxybutanoate (10d) (1.5 g, 1.55 mmol, 1.0 eq) and TEA (470.63 mg, 4.65 mmol, 647.36 μL, 3.0 eq) dissolved in dry DCM (30 mL) was added triphosgene (276.04 mg, 930.20 μmol, 0.6 eq) at 0 °C under N2. The resulting solution was stirred at 25 °C for 2 h and concentrated under reduced pressure give [8-[chlorocarbonyl-[(1- methyl-4-piperidyl)methyl]amino]-15-(4,4-diheptoxybutanoyloxy)pentadecyl] 4,4- diheptoxybutanoate (10e) (1.6 g, crude) as a yellow solid. [00364] Step 10: 8-(((heptylthio)carbonyl)((1-methylpiperidin-4- yl)methyl)amino)pentadecane-1,15-diyl bis(4,4-bis(heptyloxy)butanoate) (10)

[00365] To a solution of heptane-1-thiol (616.40 mg, 4.66 mmol, 730.33 μL, 3.0 eq) dissolved in dry THF (30 mL) was added NaOH (186.40 mg, 4.66 mmol, 3.0 eq) at 0 °C under N2. Then [8-[chlorocarbonyl-[(1-methyl-4-piperidyl)methyl]amino]-15-(4,4- diheptoxybutanoyloxy)pentadecyl] 4,4-diheptoxybutanoate (10e) (1.6 g, 1.55 mmol, 1.0 eq) dissolved in THF (10 mL) was added via syringe slowly under N2 at 0 °C. The resulting solution was stirred at 25 °C for 4 h. The reaction mixture was quenched by NH₄Cl (10 mL) at 0 °C and then diluted with EtOAc (5 mL). The aqueous phase was extracted with EtOAc (20 mL × 2). The combined organic phase was washed with brine (30 mL), dried with anhydrous sodium sulfate, filtered and concentrated in vacuum to give residue. The residue was purified by prep- HPLC (FA condition;column: CD11 - Welch Xtimate C1150 * 25 * 10 μm; mobile phase: [water (FA) - ACN]; gradient: 54% - 84% B over 10 min) to give [15-(4,4- diheptoxybutanoyloxy)-8-[heptylsulfanylcarbonyl-[(1-methyl-4- piperidyl)methyl]amino]pentadecyl] 4,4-diheptoxybutanoate (10) (367 mg, 325.96 μmol, 36.7% yield, 99.99% purity) as a yellow oil. LCMS: [M+H]⁺: 1126.1¹H NMR (400 MHz, CDCl₃) δ = 4.50 (t, J = 5.6 Hz, 2H), 4.06 (t, J = 6.8 Hz, 4H), 3.92 - 3.73 (m, 1H), 3.62 - 3.53 (m, 4H), 3.46 - 3.35 (m, 4H), 3.17 - 2.96 (m, 4H), 2.87 (t, J = 7.2 Hz, 2H), 2.38 (t, J = 7.6 Hz, 7H), 2.32 - 2.04 (m, 3H), 2.03 - 1.83 (m, 6H), 1.80 - 1.72 (m, 2H), 1.63 - 1.53 (m, 14H), 1.48 - 1.41 (m, 4H), 1.38 - 1.23 (m, 56H), 1.00 - 0.77 (m, 15H). Example 11: Synthesis of bis(2-butyloctyl) 10-(((heptylthio)carbonyl)((1-methylpiperidin-3- yl)methyl)amino)nonadecanedioate (11) [00366] Step 1: bis(2-butyloctyl) 10-(((1-methylpiperidin-3- yl)methyl)amino)nonadecanedioate (11a)

[00367] A mixture of (1-methyl-3-piperidyl)methanamine (350 mg, 2.73 mmol, 1.0 eq), NaBH(OAc)₃ (867.83 mg, 4.09 mmol, 1.5 eq), HOAc (245.89 mg, 4.09 mmol, 234.40 μL, 1.5 eq) in DCM (25 mL) was degassed and purged with N₂3 times, bis(2-butyloctyl) 10- oxononadecanedioate (1f) (2.22 g, 3.28 mmol, 1.2 eq) in DCM (10 mL) was added and then the mixture was stirred at 20 °C for 10 h under N2. The mixture pH was adjusted to 9 - 10 was with 1 M NaOH (20 mL), the aqueous phase was extracted with EtOAc (35 mL × 3). The combined organic phase was washed with brine (40 mL), dried with anhydrous sodium sulfate, filtered and concentrated in vacuum to give crude product. The residue was purified by flash silica gel chromatography (ISCO®; 20g SepaFlash® Silica Flash Column, Eluent of 0 ~ 10% MeOH / DCM gradient @ 100 mL / min) to give bis(2-butyloctyl) 10-[(1-methyl-3- piperidyl)methylamino]nonadecanedioate (11a) (1.56 g, 1.97 mmol, 72.2% yield) as a yellow oil. ¹H NMR (400 MHz, CDCl₃) δ = 3.97 (d, J = 6.0 Hz, 4H), 3.03 - 2.72 (m, 2H), 2.55 - 2.38 (m, 3H), 2.38 - 2.25 (m, 7H), 2.07 - 1.97 (m, 1H), 1.89 - 1.67 (m, 5H), 1.66 - 1.58 (m, 6H), 1.42 - 1.17 (m, 58H), 0.95 - 0.83 (m, 12H). [00368] Step 2: bis(2-butyloctyl) 10-((chlorocarbonyl)((1-methylpiperidin-3- yl)methyl)amino)nonadecanedioate (11b)

[00369] To a solution of bis(2-butyloctyl) 10-[(1-methyl-3- piperidyl)methylamino]nonadecanedioate (11a) (1.0 g, 1.26 mmol, 1.0 eq) and TEA (383.62 mg, 3.79 mmol, 527.68 μL, 3.0 eq) dissolved in dry DCM (15 mL) was added triphosgene (225.00 mg, 758.23 μmol, 0.6 eq) at 0 °C under N2. The resulting solution was stirred at 25 °C for 1 h and concentrated under reduced pressure to give bis(2-butyloctyl) 10-[chlorocarbonyl-[(1- methyl-3-piperidyl)methyl]amino]nonadecanedioate (11b) (1.08 g, crude) was obtained as a yellow solid. [00370] Step 4: bis(2-butyloctyl) 10-(((heptylthio)carbonyl)((1-methylpiperidin-3- yl)methyl)amino)nonadecanedioate (11)

[00371] To a solution of heptane-1-thiol (501.94 mg, 3.79 mmol, 594.71 μL, 3.0 eq) dissolved in dry THF (20 mL) was added NaOH (151.79 mg, 3.79 mmol, 3.0 eq) at 0 °C under N2. Then bis(2-butyloctyl) 10-[chlorocarbonyl-[(1-methyl-3-piperidyl)methyl]amino]nonadecanedioate (11b) (1.08 g, 1.26 mmol, 1.0 eq) dissolved in THF (10 mL) was added via syringe slowly under N2 at 0 °C. The resulting solution was stirred at 25 ° C for 4 h. The reaction mixture was quenched by NH₄Cl (16 mL) at 0 °C and then diluted with EtOAc (30 mL). The aqueous phase was extracted with EtOAc (30 mL × 2). The combined organic phase was washed with brine (60 mL), dried with anhydrous sodium sulfate, filtered and concentrated in vacuum to give residue. The residue was purified by flash silica gel chromatography (ISCO®; 20 g SepaFlash® Silica Flash Column, Eluent of 0 ~ 5% MeOH / DCM gradient @ 45 mL / min) to give bis(2- butyloctyl) 10-[heptylsulfanylcarbonyl-[(1-methyl-3-piperidyl)methyl]amino]nonadecanedioate (11) (1.18 g, 1.24 mmol, 98.2% yield, 99.99% purity) as a yellow oil. LCMS: [M+H]⁺: 950.4¹H NMR (400 MHz, CDCl₃) δ = 3.97 (d, J = 6.0 Hz, 4H), 3.90 - 3.67 (m, 1H), 3.20 - 3.04 (m, 2H), 2.87 (t, J = 7.2 Hz, 2H), 2.82 - 2.64 (m, 2H), 2.36 - 2.17 (m, 7H), 2.04 - 1.84 (m, 2H), 1.83 - 1.68 (m, 3H), 1.64 - 1.57 (m, 10H), 1.55 - 1.40 (m, 4H), 1.36 - 1.23 (m, 60H), 0.94 - 0.84 (m, 15H). Example 12: Synthesis of bis(2-butyloctyl) 10-(((1-methylpiperidin-4- yl)methyl)(octylsulfinyl)amino)nonadecanedioate (12)

[00372] To a solution of bis(2-butyloctyl) 10-[(1-methyl-4- piperidyl)methylamino]nonadecanedioate (5f) (1 g, 1.26 mmol, 1.0 eq), and TEA (3.20 g, 31.59 mmol, 4.40 mL, 25.0 eq) in DCM (15 mL) was added octane-1-sulfinyl chloride (7b) (994.48 mg, 5.05 mmol, 4.0 eq) in DCM (3 mL) at 0 °C. The mixture was stirred at 25 °C for 1 h. The reaction was concentrated. The residue was purified by flash silica gel chromatography (ISCO®; 20 g SepaFlash® Silica Flash Column, Eluent of 0 ~ 6% EtOAc / PE gradient @ 45 mL / min) to give bis(2-butyloctyl) 10-[(1-methyl-4-piperidyl)methyl-octylsulfinyl- amino]nonadecanedioate (12) (300 mg, 315.23 μmol, 25.0% yield, 99.99% purity) as a yellow oil. LCMS: [M+H]⁺: 952.0¹H NMR (400 MHz, CDCl3) δ = 3.97 (d, J = 6.0 Hz, 4H), 3.13 - 3.04 (m, 1H), 3.02 - 2.88 (m, 3H), 2.76 - 2.65 (m, 2H), 2.60 - 2.47 (m, 1H), 2.35 (s, 3H), 2.32 - 2.27 (m, 4H), 2.07 - 1.90 (m, 2H), 1.84 - 1.78 (m, 1H), 1.76 - 1.69 (m, 2H), 1.66 - 1.57 (m, 8H), 1.55 - 1.45 (m, 4H), 1.44 - 1.36 (m, 4H), 1.35 - 1.24 (m, 60H), 0.97 - 0.81 (m, 15H). Example 13: Synthesis of bis(2-butyloctyl) 10-[heptylsulfanylcarbonyl-[(4-methylmorpholin-2- yl)methyl]amino]nonadecanedioate (13) [00373] Step 1: bis(2-butyloctyl) 10-[(4-methylmorpholin-2- yl)methylamino]nonadecanedioate (13a):

[00374] A mixture of (4-methylmorpholin-2-yl)methanamine (460.09 mg, 3.53 mmol, 1.2 eq) and NaBH(OAc)₃ (936.26 mg, 4.42 mmol, 1.5 eq), HOAc (265.28 mg, 4.42 mmol, 252.88 μL, 1.5 eq) in DCM (30 mL) was stirred at 25 °C for 1 h, and then bis(2-butyloctyl) 10- oxononadecanedioate (1f) (2 g, 2.95 mmol, 1 eq) was added. The resulting mixture was stirred at 25 °C for 15 h. The reaction mixture was diluted with H₂O (50 mL) and the mixture pH was adjusted to 9-10 with saturated aq. Na2CO3, the aqueous phase was extracted with DCM (50 mL × 3). The combined organic phase was washed with brine (50 mL), dried with anhydrous Na₂SO₄, filtered and concentrated under reduced pressure to give a residue. The residue was purified by flash silica gel chromatography (20 g SepaFlash® Silica Flash Column, DCM : MeOH: 0~10%, 1% NH3•H2O in MeOH) to afford bis(2-butyloctyl) 10-[(4-methylmorpholin-2- yl)methylamino]nonadecanedioate (13a) (1.3 g, 1.63 mmol, 55.5% yield, 99.7% purity) as yellow oil. LCMS: [M+H]⁺: 793.9 [00375] Step 2: bis(2-butyloctyl) 10-[chlorocarbonyl-[(4-methylmorpholin-2- yl)methyl]amino]nonadecanedioate (13b)

[00376] To a solution of bis(2-butyloctyl) 10-[(4-methylmorpholin-2- yl)methylamino]nonadecanedioate (13a) (1.2 g, 1.51 mmol, 1 eq) and TEA (306.14 mg, 3.03 mmol, 421.10 μL, 2 eq) dissolved in dry DCM (20 mL) was added triphosgene (269.33 mg, 907.61 μmol, 0.6 eq) at 0 °C under N2. The resulting solution was stirred at 20 °C for 2 h. The reaction mixture was concentrated under reduced pressure to give bis(2-butyloctyl) 10- [chlorocarbonyl-[(4-methylmorpholin-2-yl)methyl]amino]nonadecanedioate (13b) (1.3 g, crude) as a yellow solid. The crude product was used for next step starting material. [00377] Step 3: bis(2-butyloctyl) 10-[heptylsulfanylcarbonyl-[(4-methylmorpholin-2- yl)methyl]amino]nonadecanedioate (13)

[00378] To a solution of heptane-1-thiol (602.79 mg, 4.56 mmol, 714.21 μL, 3 eq) dissolved in dry THF (10 mL) was added NaOH (182.30 mg, 4.56 mmol, 3 eq) at 0 °C under N2. After addition, the resulting solution was stirred at 0 °C for 0.5 h, and then bis(2-butyloctyl) 10- [chlorocarbonyl-[(4-methylmorpholin-2-yl)methyl]amino]nonadecanedioate (13b) (1.3 g, 1.52 mmol, 1 eq) in THF (10 mL) was added dropwise. The resulting solution was stirred at 20 °C for 15.5 h. The reaction mixture was quenched by NH₄Cl (50 mL) at 0 °C and then diluted with DCM (30 mL). The aqueous phase was extracted with DCM (30 mL × 3). The combined organic phase was washed with brine (60 mL), dried with anhydrous Na2SO4, filtered and concentrated in vacuum to give residue. The residue was purified by flash silica gel chromatography (40 g SepaFlash® Silica Flash Column, DCM : MeOH: 0~10%) and prep- HPLC (column: CD11-Welch Xtimate C1150 × 25 × 10um; mobile phase: [water (FA) -ACN]; gradient: 54% - 84% B over 10 min) to give bis(2-butyloctyl) 10-[heptylsulfanylcarbonyl-[(4- methylmorpholin-2-yl)methyl]amino]nonadecanedioate (13) (680 mg, 714.55 μmol, 48.6% yield, 99.99% purity) as yellow oil. LCMS: [M+H]⁺: 952.0¹H NMR (400 MHz, CD3OD-d4) δ = 4.00 (d, J = 5.6 Hz, 4H), 3.85 (d, J = 9.6 Hz, 2H), 3.76 - 3.72 (m, 1H), 3.58 - 3.50 (m, 1H), 3.41 - 3.33 (m, 1H), 3.21 (q, J = 7.2 Hz, 1H), 3.03 - 2.95 (m, 1H), 2.89 (t, J = 7.2 Hz, 2H), 2.79 - 2.62 (m, 3H), 2.39 - 2.27 (m, 7H), 2.26 - 2.16 (m, 1H), 2.00 - 1.90 (m, 1H), 1.71 - 1.58 (m, 10H), 1.38 - 1.28 (m, 60H), 0.95 - 0.89 (m, 15H). Example 14: Synthesis of bis(2-butyloctyl) 10-(N-((1-methylpiperidin-4- yl)methyl)octylsulfonamido)nonadecanedioate (14)

[00379] NaIO₄ (53.94 mg, 252.21 μmol, 13.98 μL, 1.2 eq) and RuCl₃ (8.72 mg, 42.03 μmol, 2.80 μL, 0.2 eq) was added to a 0 °C solution of bis(2-butyloctyl) 10-[(1-methyl-4- piperidyl)methyl-octylsulfinyl amino]nonadecanedioate (12) (200 mg, 210.17 μmol, 1.0 eq) in DCM (1 mL), MeCN (1 mL) and H2O (1.6 mL) and stirred for 1 h. After completely, DCM (5 mL) was added and separated water phase, and exacted with DCM (3 mL × 3), washed with brine (5 mL), dried over anhydrous sodium sulfate, filtered and concentrated under reduced pressure to give a residue. The residue was purified by flash silica gel chromatography (ISCO®; 4 g SepaFlash® Silica Flash Column, Eluent of 0 ~ 5% MeOH / DCM gradient @ 15 mL / min) to give bis(2-butyloctyl) 10-[(1-methyl-4-piperidyl)methyl-octylsulfonyl- amino]nonadecanedioate (14) (55 mg, 56.84 μmol, 27.04% yield, 99.99% purity) was obtained as a yellow oil. LCMS: [M+H]⁺: 967.9¹H NMR (400 MHz, CD₃OD-d₄) δ = 4.00 (d, J = 5.6 Hz, 4H), 3.55 - 3.45 (m, 1H), 3.44 - 3.35 (m, 2H), 3.13 - 3.04 (m, 2H), 3.02 - 2.93 (m, 2H), 2.89 - 2.79 (m, 2H), 2.76 (s, 3H), 2.36 - 2.23 (m, 4H), 2.07 - 1.95 (m, 2H), 1.94 - 1.85 (m, 1H), 1.82 - 1.71 (m, 2H), 1.68 - 1.52 (m, 10H), 1.49 - 1.42 (m, 4H), 1.40-1.25 (m, 60H), 0.99 - 0.78 (m, 15H). Example 15: Synthesis of bis(2-butyloctyl) 10-(((heptylthio)carbonyl)((1-(2- hydroxyethyl)piperidin-4-yl)methyl)amino)nonadecanedioate (15) [00380] Step 1: (1-(2-((tert-butyldimethylsilyl)oxy)ethyl)piperidin-4-yl)methanamine (15a)

[00381] A mixture of 2-[4-(aminomethyl)-1-piperidyl]ethanol (2 g, 12.64 mmol, 1.0 eq), imidazole (1.03 g, 15.17 mmol, 1.2 eq) in DCM (40 mL) was degassed and purged with N23 times, then TBSCl (2.29 g, 15.17 mmol, 1.87 mL, 1.2 eq) was added, and then the mixture was stirred at 25 °C for 10 h under N2. The reaction mixture was diluted with H2O 15 mL and extracted with DCM 30 mL (15 mL × 2). The combined organic layers were washed with brine 30 mL (15 mL × 2), dried over anhydrous sodium sulfate, filtered and concentrated under reduced pressure to give a residue. The residue was purified by flash silica gel chromatography (ISCO®; 20 g SepaFlash® Silica Flash Column, Eluent of 0 ~ 15% EtOAc / PE gradient @ 100 mL / min) to give [1-[2-[tert-butyl(dimethyl)silyl]oxyethyl]-4-piperidyl]methanamine (15a) (1.71 g, 5.96 mmol, 47.2% yield, 95% purity) as a colorless oil. [00382] Step 2: bis(2-butyloctyl) 10-(((1-(2-((tert-butyldimethylsilyl)oxy)ethyl)piperidin-4- yl)methyl)amino)nonadecanedioate (15b):

[00383] A mixture of [1-[2-[tert-butyl(dimethyl)silyl]oxyethyl]-4-piperidyl]methanamine (15a) (1.0 g, 3.67 mmol, 1.0 eq), NaBH(OAc)3 (1.17 g, 5.50 mmol, 1.5 eq), HOAc (330.56 mg, 5.50 mmol, 315.12 μL, 1.5 eq) in DCM (35 mL) was degassed and purged with N23 times, bis(2-butyloctyl) 10-oxononadecanedioate (1f) (2.99 g, 4.40 mmol, 1.2 eq) in DCM (8 mL) was added and then the mixture was stirred at 20 °C for 10 h under N2. The mixture pH was adjusted to 9 - 10 was with 1 M NaOH (20 mL), the aqueous phase was extracted with EtOAc (35 mL × 3). The combined organic phase was washed with brine (40 mL), dried with anhydrous sodium sulfate, filtered and concentrated in vacuum to give crude product. The residue was purified by flash silica gel chromatography (ISCO®; 20 g SepaFlash® Silica Flash Column, Eluent of 0 ~ 10% MeOH / DCM gradient @ 100 mL / min) to give bis(2-butyloctyl) 10-[[1-[2-[tert- butyl(dimethyl)silyl]oxyethyl]-4-piperidyl]methylamino]nonadecanedioate (15b) (2.58 g, 2.54 mmol, 69.1% yield, 92% purity) as a yellow oil. ¹H NMR (400 MHz, CDCl3) δ = 3.97 (d, J = 6.0 Hz, 4H), 3.78 (t, J = 6.4 Hz, 2H), 3.07 - 2.92 (m, 2H), 2.62 - 2.51 (m, 2H), 2.49 - 2.41 (m, 3H), 2.29 (t, J = 7.6 Hz, 4H), 2.18 - 2.06 (m, 2H), 1.77 - 1.70 (m, 2H), 1.65 - 1.58 (m, 6H), 1.34 - 1.24 (m, 60H), 0.91 - 0.86 (m, 21H), 0.06 (s, 6H). [00384] Step 3: bis(2-butyloctyl) 10-(((1-(2-((tert-butyldimethylsilyl)oxy)ethyl)piperidin-4- yl)methyl)(chlorocarbonyl)amino)nonadecanedioate (15c)

[00385] To a solution of bis(2-butyloctyl)10-[[1-[2-[tert-butyl(dimethyl)silyl]oxyethyl]-4- piperidyl]methylamino]nonadecanedioate (15b) (2.09 g, 2.23 mmol, 1.0 eq) and TEA (678.12 mg, 6.70 mmol, 932.77 μL, 3.0 eq) dissolved in dry DCM (30 mL) was added triphosgene (397.73 mg, 1.34 mmol, 0.6 eq) at 0 °C under N2. The resulting solution was stirred at 25 °C for 2 h and concentrated under reduced pressure and kept under N2. The reaction mixture was concentrated in vacuum to give bis(2-butyloctyl) 10-[[1-[2-[tert-butyl(dimethyl)silyl]oxyethyl]- 4-piperidyl]methyl-chlorocarbonyl-amino]nonadecanedioate (15c) (2.23 g, crude) as a yellow solid. [00386] Step 4: bis(2-butyloctyl) 10-(((1-(2-((tert-butyldimethylsilyl)oxy)ethyl)piperidin-4- yl)methyl)((heptylthio)carbonyl)amino)nonadecanedioate (15d)

[00387] To a solution of heptane-1-thiol (591.05 mg, 4.47 mmol, 700.30 μL, 2.0 eq) dissolved in dry THF (30 mL) was added NaOH (1.07 g, 26.81 mmol, 12.0 eq) at 0 °C under N₂. bis(2- butyloctyl)10-[[1-[2-[tert-butyl(dimethyl)silyl]oxyethyl]-4-piperidyl]methyl-chlorocarbonyl- amino]nonadecanedioate (15c) (2.23 g, 2.23 mmol, 1.0 eq) dissolved in THF (10 mL) was added via syringe slowly under N₂ at 0 °C. The resulting solution was stirred at 25 ° C for 15 h. The reaction mixture was quenched by NH4Cl (16 mL) at 0 °C and then diluted with DCM (30 mL). The aqueous phase was extracted with EtOAc (30 mL × 2). The combined organic phase was washed with brine (60 mL), dried with anhydrous sodium sulfate, filtered and concentrated in vacuum to give residue. The residue was purified by flash silica gel chromatography (ISCO®; 20 g SepaFlash® Silica Flash Column, Eluent of 0 ~ 10% MeOH / DCM gradient @ 80 mL / min) to give bis(2-butyloctyl) 10-[[1-[2-[tert-butyl(dimethyl)silyl]oxyethyl]-4-piperidyl]methyl- heptylsulfanylcarbonyl-amino]nonadecanedioate (15d) (1.68 g, 1.46 mmol, 65.30% yield, 95% purity) as a yellow oil.¹H NMR (400 MHz, CDCl3) δ = 3.97 (d, J = 6.0 Hz, 4H), 3.90 - 3.67 (m, 3H), 3.16 - 2.90 (m, 4H), 2.90 - 2.81 (m, 2H), 2.63 - 2.42 (m, 2H), 2.34 - 2.25 (m, 4H), 2.06 - 1.95 (m, 1H), 1.67 - 1.56 (m, 10H), 1.46 - 1.38 (m, 4H), 1.37 - 1.19 (m, 64H), 0.94 - 0.84 (m, 24H), 0.06 (s, 6H). [00388] Step 5: bis(2-butyloctyl) 10-(((heptylthio)carbonyl)((1-(2-hydroxyethyl)piperidin-4- yl)methyl)amino)nonadecanedioate (15)

[00389] A mixture of bis(2-butyloctyl) 10-[[1-[2-[tert-butyl(dimethyl)silyl]oxyethyl]-4- piperidyl]methyl-heptylsulfanylcarbonyl-amino]nonadecanedioate (15d) (0.8 g, 731.35 μmol, 1.0 eq) in THF (15 mL) , then TBAF (1 M, 1.10 mL, 1.5 eq) was added slowly at 0 °C, the mixture was degassed and purged with N23 times, and then the mixture was stirred at 20 °C for 2 h under N2. The reaction mixture was quenched by addition H2O (15 mL) at 0 °C, and then diluted with EtOAc (5 mL) and extracted with EtOAc 30 mL (10 mL × 3). The combined organic layers were washed with brine 30 mL, dried over anhydrous sodium sulfate, filtered and concentrated under reduced pressure to give a residue. The residue was purified by flash silica gel chromatography (ISCO®; 12 g SepaFlash® Silica Flash Column, Eluent of 0 ~ 5% MeOH / DCM gradient @ 15 mL / min) to give bis(2-butyloctyl) 10-[heptylsulfanylcarbonyl-[[1-(2- hydroxyethyl)-4-piperidyl]methyl]amino]nonadecanedioate (15) (440 mg, 439.28 μmol, 60.1% yield, 97.8% purity) as a yellow oil. LCMS: [M+H]⁺: 980.0¹H NMR (400 MHz, CDCl₃) δ = 3.97 (d, J = 6.0 Hz, 4H), 3.88 - 3.72 (m, 1H), 3.71 - 3.59 (m, 2H), 3.17 - 2.96 (m, 4H), 2.87 (t, J = 7.2 Hz, 2H), 2.67 - 2.55 (m, 2H), 2.30 (t, J = 7.6 Hz, 4H), 2.26 - 2.00 (m, 3H), 1.93 - 1.78 (m, 1H), 1.76 - 1.69 (m, 2H), 1.65 - 1.57 (m, 8H), 1.47 - 1.36 (m, 6H), 1.34 - 1.21 (m, 60H), 0.94 - 0.83 (m, 15H). Example 16: Synthesis of bis(2-butyloctyl) 10-(N-((1-methylpiperidin-3- yl)methyl)octylsulfonamido)nonadecanedioate (16) [00390] Step 1: bis(2-butyloctyl) 10-(((1-methylpiperidin-3- yl)methyl)(octylsulfinyl)amino)nonadecanedioate (16a)

[00391] To a solution of bis(2-butyloctyl) 10-[(1-methyl-3- piperidyl)methylamino]nonadecanedioate (11a) (150 mg, 189.56 μmol, 1.0 eq) and TEA (383.62 mg, 3.79 mmol, 527.68 μL, 20.0 eq) in DCM (5 mL) was added octane-1-sulfinyl chloride (223.76 mg, 1.14 mmol, 6.0 eq) in DCM (3 mL) at 0 °C. The mixture was stirred at 20 °C for 2 h. The reaction was concentrated and the residue was purified by flash silica gel chromatography (ISCO®; 12 g SepaFlash® Silica Flash Column, Eluent of 0 ~ 10% MeOH / DCM gradient @ 15 mL / min) to give bis(2-butyloctyl) 10-[(1-methyl-3-piperidyl)methyl-octylsulfinyl- amino]nonadecanedioate (16a) (175 mg, 176.55 μmol, 93.1% yield, 96% purity) as a yellow oil. ¹H NMR (400 MHz, CDCl₃) δ = 3.97 (d, J = 6.0 Hz, 4H), 3.75 - 3.25 (m, 2H), 3.19 - 2.89 (m, 3H), 2.87 - 2.78 (m, 1H), 2.77 - 2.63 (m, 3H), 2.62 - 2.39 (m, 2H), 2.36 - 2.20 (m, 5H), 2.08 - 1.81 (m, 4H), 1.78 - 1.62 (m, 8H), 1.54 - 1.40 (m, 6H), 1.34 - 1.24 (m, 60H), 0.91 - 0.85 (m, 15H). [00392] Step 2: bis(2-butyloctyl) 10-(N-((1-methylpiperidin-3- yl)methyl)octylsulfonamido)nonadecanedioate (16)

[00393] To a solution of bis(2-butyloctyl) 10-[(1-methyl-3-piperidyl)methyl-octylsulfinyl- amino]nonadecanedioate (16a) (160 mg, 168.14 μmol, 1.0 eq) in DCM (1 mL) ,MeCN (1 mL) and H₂O (1.6 mL) were added NaIO₄ (43.16 mg, 201.77 μmol, 11.18 μL, 1.2 eq) and RuCl₃ (6.98 mg, 33.63 μmol, 2.24 μL, 0.2 eq) at 0 °C and stirred for 20 min. after completely, DCM (5 mL) was added and separated water phase, and exacted with DCM (3 mL × 3), washed with brine (5 mL), dried over anhydrous sodium sulfate, filtered and concentrated under reduced pressure to give a residue. The residue was purified by flash silica gel chromatography (ISCO®; 4 g SepaFlash® Silica Flash Column, Eluent of 0 ~ 5% MeOH / DCM gradient @ 15 mL / min).then was further purified by prep-HPLC (FA condition; column: CD11-Welch Xtimate C1 150 × 25 × 10 μm; mobile phase: [water (FA) - ACN]; gradient: 50% - 80% B over 10 min) to give bis(2-butyloctyl) 10-[(1-methyl-3-piperidyl)methyl-octylsulfonyl-amino]nonadecanedioate (16) (55 mg, 55.72 μmol, 33.14% yield, 98.02% purity) as a yellow oil. LCMS: [M+H]⁺: 967.9 ¹H NMR (400 MHz, CDCl₃) δ = 3.97 (d, J = 6.0 Hz, 4H), 3.53 - 3.39 (m, 1H), 3.14 - 2.95 (m, 2H), 2.90 - 2.83 (m, 2H), 2.47 - 2.19 (m, 7H), 2.12 - 1.85 (m, 3H), 1.85 - 1.68 (m, 6H), 1.66 - 1.59 (m, 6H), 1.57 - 1.47 (m, 4H), 1.46 - 1.36 (m, 4H), 1.36 - 1.20 (m, 60H), 0.97 - 0.83 (m, 15H). Example 17: Synthesis of bis(2-butyloctyl) 10-(N-((1-methylpiperidin-4-yl)methyl)decan-3- ylsulfonamido)nonadecanedioate (17) [00394] Step 1: decan-3-yl 4-methylbenzenesulfonate (17a)

[00395] A mixture of decan-3-ol (10 g, 63.18 mmol, 1.0 eq), TEA (9.59 g, 94.77 mmol, 13.19 mL, 1.5 eq) in DCM (200 mL) was degassed and purged with N₂3 times, Then MsCl (9.41 g, 82.13 mmol, 6.36 mL, 1.3 eq) was added dropwise at 0 °C, and then the mixture was stirred at 25 °C for 1 h under N2. The reaction mixture was quenched by addition NH4Cl (30 mL) at 0 °C, diluted with H₂O (50 mL) and extracted with DCM 300 mL (100 mL × 3). The combined organic layers were washed with brine 300 mL (100 mL × 3), dried over anhydrous sodium sulfate, filtered and concentrated under reduced pressure to give a residue. The residue was purified by flash silica gel chromatography (ISCO®; 120 g SepaFlash® Silica Flash Column, Eluent of 0 ~ 30% EtOAc / PE gradient @ 100 mL / min) to give 1-ethyloctyl methanesulfonate (17a) (15 g, 63.46 mmol, 100.0% yield) as a yellow oil. ¹H NMR (400 MHz, CDCl3) δ = 4.71 - 4.58 (m, 1H), 3.00 (s, 3H), 1.81 - 1.62 (m, 4H), 1.43 - 1.21 (m, 10H), 0.98 (t, J = 7.2 Hz, 3H), 0.88 (t, J = 6.8 Hz, 3H). [00396] Step 2: S-(decan-3-yl) ethanethioate (17b)

[00397] A mixture of 1-ethyloctyl methanesulfonate (17a) (7 g, 29.61 mmol, 1.0 eq), thioacetic acid (2.71 g, 35.54 mmol, 2.54 mL, 1.2 eq), Cs₂CO₃ (11.58 g, 35.54 mmol, 1.2 eq) in DMF (50 mL) was degassed and purged with N23 times, and then the mixture was stirred at 60 °C for 16 h under N2. The reaction mixture was diluted with H2O (100 mL) and extracted with EtOAc 300 mL (100 mL × 3). The combined organic layers were washed with brine 300 mL (100 mL × 3), dried over anhydrous sodium sulfate, filtered and concentrated under reduced pressure to give a residue. The residue was purified by flash silica gel chromatography (ISCO®; 80 g SepaFlash® Silica Flash Column, Eluent of 0 ~ 10% EtOAc / PE gradient @ 100 mL / min) to give S-(1-ethyloctyl) ethanethioate (17b) (4.6 g, 19.56 mmol, 70.5% yield, 92% purity) as a white solid. ¹H NMR (400 MHz, CDCl3) δ = 3.54 - 3.38 (m, 1H), 2.32 (s, 3H), 1.65 - 1.47 (m, 4H), 1.33 - 1.21 (m, 10H), 0.94 (t, J = 7.2 Hz, 3H), 0.88 (t, J = 6.8 Hz, 3H). [00398] Step 3: decane-3-sulfinic chloride (17c)

[00399] A mixture of S-(1-ethyloctyl) ethanethioate (17b) (3.47 g, 16.04 mmol, 1.0 eq) in DCM (40 mL) was degassed and purged with N₂3 times and cool down to 0 °C, the Ac₂O (1.64 g, 16.04 mmol, 1.51 mL, 1.0 eq) and sulfuryl chloride (4.33 g, 32.07 mmol, 3.21 mL, 2.0 eq) in DCM (3 mL) was added slowly via injection, then the mixture was stirred at 0 °C for 1 h under N₂. The reaction mixture was concentrated under reduced pressure (30 °C) to give decane-3- sulfinyl chloride (17c) (3.68 g, crude) as a light yellow oil. [00400] Step 4: bis(2-butyloctyl) 10-((decan-3-ylsulfinyl)((1-methylpiperidin-4- yl)methyl)amino)nonadecanedioate (17d)

[00401] To a solution of bis(2-butyloctyl) 10-[(1-methyl-4- piperidyl)methylamino]nonadecanedioate (5f) (2 g, 2.53 mmol, 1.0 eq), DMAP (61.75 mg, 505.48 μmol, 0.2 eq) and TEA (5.11 g, 50.55 mmol, 7.04 mL, 20.0 eq) in DCM (20 mL) was added decane-3-sulfinyl chloride (17c) (2.84 g, 12.64 mmol, 5.0 eq) in DCM (10 mL) at 0 °C. The mixture was stirred at 25 °C for 1 h. The reaction mixture was diluted with H2O (30 mL) and extracted with DCM 30 mL (10 mL × 3). The combined organic layers were washed with brine 30 mL (10 mL × 3), dried over anhydrous sodium sulfate, filtered and concentrated under reduced pressure to give a residue. The residue was purified by flash silica gel chromatography (ISCO®;40 g SepaFlash® Silica Flash Column, Eluent of 0 ~ 10% MeOH / DCM gradient @ 100 mL / min) to give bis(2-butyloctyl) 10-[1-ethyloctylsulfinyl-[(1-methyl-4- piperidyl)methyl]amino]nonadecanedioate (17d) (931 mg, 940.84 μmol, 37.2% yield, 99% purity) as a yellow oil.¹H NMR (400 MHz, CDCl3) δ = 3.97 (d, J = 6.0 Hz, 4H), 3.14 - 2.77 (m, 4H), 2.71 - 2.49 (m, 2H), 2.45 - 2.18 (m, 7H), 2.10 - 1.88 (m, 2H), 1.87 - 1.69 (m, 5H), 1.67 - 1.56 (m, 8H), 1.51 - 1.39 (m, 6H), 1.38 - 1.13 (m, 62H), 1.04 - 0.75 (m, 18H). [00402] Step 5: bis(2-butyloctyl) 10-(N-((1-methylpiperidin-4-yl)methyl)decan-3- ylsulfonamido)nonadecanedioate (17)

[00403] To a solution of bis(2-butyloctyl) 10-[1-ethyloctylsulfinyl-[(1-methyl-4- piperidyl)methyl]amino]nonadecanedioate (17d) (100.00 mg, 102.08 μmol, 1.0 eq) in DCM (1 mL), MeCN (1 mL) and H2O (1.6 mL) were added NaIO4 (26.20 mg, 122.49 μmol, 6.79 μL, 1.2 eq) and RuCl3 (4.23 mg, 20.42 μmol, 1.36 μL, 0.2 eq) at 0 °C and stirred for 20 min. after completely, DCM (25 mL) was added and separated water phase, and exacted with DCM (10 mL × 3), washed with brine (30 mL), dried over anhydrous sodium sulfate, filtered and concentrated under reduced pressure to give a residue. The residue was purified by flash silica gel chromatography (ISCO®; 20 g SepaFlash® Silica Flash Column, Eluent of 0 ~ 5% MeOH / DCM gradient @ 40 mL / min) to give bis(2-butyloctyl) 10-[1-ethyloctylsulfonyl-[(1-methyl-4- piperidyl)methyl]amino]nonadecanedioate (17) (140 mg, 138.56 μmol, 5.52% yield, 98.54% purity) as a yellow oil. LCMS: [M+H]⁺: 995.6¹H NMR (400 MHz, CDCl3) δ = 3.97 (d, J = 6.0 Hz, 4H), 3.42 - 3.34 (m, 1H), 3.33 - 3.11 (m, 2H), 3.10 - 2.97 (m, 2H), 2.80 - 2.71 (m, 1H), 2.59 (s, 3H), 2.30 (t, J = 7.6 Hz, 4H), 2.05 - 1.87 (m, 2H), 1.82 - 1.69 (m, 5H), 1.67 - 1.53 (m, 8H), 1.51 - 1.43 (m, 4H), 1.41 - 1.17 (m, 64H), 1.03 (t, J = 7.2 Hz, 3H), 0.94 - 0.82 (m, 15H). Example 18: Synthesis of (S)-8-(((1-methylpiperidin-4- yl)methyl)(octylsulfinyl)amino)pentadecane-1,15-diyl bis(4,4-bis(pentyloxy)butanoate)/ (R)-8- (((1-methylpiperidin-4-yl)methyl)(octylsulfinyl)amino)pentadecane-1,15-diyl bis(4,4- bis(pentyloxy)butanoate) (18R, 18S) [00404] Step 1: 8-(((1-methylpiperidin-4-yl)methyl)(octylsulfinyl)amino)pentadecane-1,15- diyl bis(4,4-bis(pentyloxy)butanoate) (18a)

[00405] To a solution of [15-(4,4-dipentoxybutanoyloxy)-8-[(1-methyl-4- piperidyl)methylamino]pentadecyl] 4,4-dipentoxybutanoate (9i) (502 mg, 586.92 μmol, 1.0 eq), DMAP (14.34 mg, 117.38 μmol, 0.2 eq) and TEA (593.89 mg, 5.87 mmol, 816.91 μL, 10.0 eq) in DCM (5 mL) was added octane-1-sulfinyl chloride (7b) (461.87 mg, 2.35 mmol, 4.0 eq) in DCM (3 mL) at 0 °C. The mixture was stirred at 25 °C for 1 h. The reaction mixture was was diluted with H2O (10 mL) and extracted with DCM 15 mL (5 mL × 3). The combined organic layers were washed with brine 30 mL (10 mL × 3), dried over anhydrous sodium sulfate, filtered and concentrated under reduced pressure to give a residue. The residue was purified by flash silica gel chromatography (ISCO®; 12 g SepaFlash® Silica Flash Column, Eluent of 0 ~ 10% MeOH / DCM gradient @ 20 mL / min) to give [15-(4,4-dipentoxybutanoyloxy)-8-[(1-methyl- 4-piperidyl)methyl-[(S)-octylsulfinyl]amino]pentadecyl] 4,4-dipentoxybutanoate (18a) (390 mg, 384.0 μmol, 65.7% yield, 99% purity) as a yellow oil. [00406] Step 2: (S)-8-(((1-methylpiperidin-4-yl)methyl)(octylsulfinyl)amino)pentadecane- 1,15-diyl bis(4,4-bis(pentyloxy)butanoate)/ (R)-8-(((1-methylpiperidin-4- yl)methyl)(octylsulfinyl)amino)pentadecane-1,15-diyl bis(4,4-bis(pentyloxy)butanoate) (18R, 18S)

[00407] The [15-(4,4-dipentoxybutanoyloxy)-8-[(1-methyl-4-piperidyl)methyl-[(S)- octylsulfinyl]amino]pentadecyl] 4,4-dipentoxybutanoate (18a) (200 mg, 196.93 μmol,1.0 eq) was separated by SFC (condition: column: DAICEL CHIRALCEL OX (250 mm × 30 mm, 10 μm); mobile phase: [CO₂ - MeOH (0.1%NH₃H₂O)]; B%: 45%, isocratic elution mode) to give [15-(4,4-dipentoxybutanoyloxy)-8-[(1-methyl-4-piperidyl)methyl-[(S)- octylsulfinyl]amino]pentadecyl] 4,4-dipentoxybutanoate (18S) (79 mg, 77.30 μmol, 39.5% yield, 99.38% purity) as a yellow oil. LCMS: [M+H]⁺: 1015.6¹H NMR (400 MHz, CDCl₃) δ = 4.50 (t, J = 5.6 Hz, 2H), 4.12 - 3.97 (m, 4H), 3.64 - 3.53 (m, 4H), 3.47 - 3.36 (m, 4H), 3.15 - 2.89 (m, 4H), 2.78 - 2.66 (m, 2H), 2.60 - 2.50 (m, 1H), 2.46 - 2.30 (m, 7H), 2.16 - 2.00 (m, 2H), 1.96 - 1.90 (m, 4H), 1.86 - 1.81 (m, 1H), 1.77 - 1.69 (m, 2H), 1.66 - 1.51 (m, 16H), 1.44 - 1.18 (m, 46H), 0.96 - 0.83 (m, 15H). [00408] And [15-(4,4-dipentoxybutanoyloxy)-8-[(1-methyl-4-piperidyl)methyl-[(R)- octylsulfinyl]amino]pentadecyl] 4,4-dipentoxybutanoate (18R) (45 mg, 44.01 μmol, 22.5% yield, 99.33% purity) as a yellow oil. LCMS: [M+H]⁺: 1015.7¹H NMR (400 MHz, CDCl₃) δ = 4.50 (t, J = 5.6 Hz, 2H), 4.10 - 4.00 (m, 4H), 3.62 - 3.53 (m, 4H), 3.46 - 3.37 (m, 4H), 3.15 - 2.91 (m, 4H), 2.79 - 2.65 (m, 2H), 2.59 - 2.49 (m, 1H), 2.46 - 2.32 (m, 7H), 2.14 - 2.00 (m, 2H), 1.97 - 1.90 (m, 4H), 1.86 - 1.69 (m, 4H), 1.67 - 1.51 (m, 16H), 1.44 - 1.22 (m, 46H), 0.96 - 0.85 (m, 15H). Example 19: Synthesis of bis(6,6-bis(pentyloxy)hexyl) 6-(((heptylthio)carbonyl)((1- methylpiperidin-4-yl)methyl)amino)undecanedioate (19) [00409] Step 1: 6,6-bis(pentyloxy)hexanenitrile (19a) HO

[00410] A mixture of 6,6-dimethoxyhexanenitrile (12.2 g, 77.60 mmol, 1.0 eq) and PPTS (9.75 g, 38.80 mmol, 0.5 eq) inpentan-1-ol (34.20 g, 388.02 mmol, 42.17 mL, 5.0 eq) was degassed and purged with N23 times, and then the mixture was stirred at 110 °C for 16 h under N₂. The reaction was concentrated. The residue was purified by flash silica gel chromatography (ISCO®; 120 g SepaFlash® Silica Flash Column, Eluent of 0 ~ 3% EtOAc / PE gradient @ 100 mL / min) to give 6,6-dipentoxyhexanenitrile (19a) (12.7 g, 47.14 mmol, 84.7% yield) as a yellow oil. ¹H NMR (400 MHz, CDCl₃) δ = 4.45 (t, J = 5.6 Hz, 1H), 3.60 - 3.48 (m, 2H), 3.45 - 3.33 (m, 2H), 2.38 - 2.24 (m, 2H), 1.65 - 1.56 (m, 6H), 1.41 - 1.27 (m, 12H), 0.92 - 0.88 (m, 6H). [00411] Step 2: 6,6-bis(pentyloxy)hexanoic acid (19b)

, 1_{9a 19b} [00412] A mixture of 6,6-dipentoxyhexanenitrile (19a) (12.7 g, 47.14 mmol, 1.0 eq) and KOH (7.93 g, 141.41 mmol, 3.0 eq) in EtOH (80 mL) and H2O (80 mL) was degassed and purged with N23 times, and then the mixture was stirred at 110 °C for 5 h under N2. The reaction mixture cooled down to room temperature and concentrated, then diluted with EtOAc (200 mL), pH adjust to pH = 5 by addition 1N HCl, and extracted with EtOAc 200 mL (100 mL × 2). The combined organic layers were washed with brine (200 mL), dried over anhydrous sodium sulfate, filtered and concentrated under reduced pressure to give a residue. The residue was purified by flash silica gel chromatography (ISCO®; 120 g SepaFlash® Silica Flash Column, Eluent of 0 ~ 30% EtOAc / PE gradient @ 100 mL / min) to give 6,6-dipentoxyhexanoic acid (19b) (7.22 g, 25.03 mmol, 53.11% yield) as a yellow oil. ¹H NMR (400 MHz, CDCl3) δ = 4.47 (t, J = 5.6 Hz, 1H), 3.61 - 3.51 (m, 2H), 3.46 - 3.33 (m, 2H), 2.36 (t, J = 7.6 Hz, 2H), 1.72 - 1.55 (m, 8H), 1.46 - 1.37 (m, 2H), 1.36 - 1.26 (m, 8H), 0.96 - 0.83 (m, 6H). [00413] Step 3: 6,6-bis(pentyloxy)hexan-1-ol (19c)

[00414] A mixture of 6,6-dipentoxyhexanoic acid (19b) (3 g, 10.40 mmol, 1.0 eq) in THF (35 mL) was degassed and purged with N23 times, Then BMS (10 M, 1.56 mL, 1.5 eq) was added dropwise at 0 °C, and then the mixture was stirred at 20 °C for 3 h under N₂. The reaction mixture was quenched by addition H₂O (30 mL) at 0 °C, and then diluted with EtOAc (50 mL) and extracted with EtOAc 90 mL (30 mL × 3). The combined organic layers were washed with brine 150 mL (50 mL × 3), dried over anhydrous sodium sulfate, filtered and concentrated under reduced pressure to give a residue. The residue was purified by flash silica gel chromatography (ISCO®; 40 g SepaFlash® Silica Flash Column, Eluent of 0 ~ 25% EtOAc / PE gradient @ 100 mL / min) to give 6,6-dipentoxyhexan-1-ol (19c) (2.59 g, 9.44 mmol, 90.73% yield) as a light yellow oil. ¹H NMR (400 MHz, DMSO-d₆) δ = 4.40 (t, J = 5.6 Hz, 1H), 4.32 (t, J = 5.2 Hz, 1H), 3.52 - 3.43 (m, 2H), 3.41 - 3.30 (m, 4H), 1.53 - 1.43 (m, 6H), 1.42 - 1.36 (m, 2H), 1.32 - 1.22 (m, 12H), 0.86 (t, J = 6.4 Hz, 6H). [00415] Step 4: dimethyl 6-isocyano-6-(p-tolylsulfonyl)undecanedioate (19d)

[00416] To a 2000 mL three neck round bottom flask under N₂, was added DMSO (1000 mL), followed by the addition of NaH (18.95 g, 473.78 mmol, 60% purity, 2.5 eq) in portions over a period of 0.5 h. The mixture was allowed to stir for 0.5 h at 20 °C following the addition, then 1- (isocyanomethylsulfonyl)-4-methyl-benzene (37 g, 189.51 mmol, 1.0 eq) was added in portions over 0.5 h, followed by the addition of TBAI (7.00 g, 18.95 mmol, 0.1 eq) in one portion. The resulting mixture was stirred for 0.5 h at 20 °C, then methyl 5-bromopentanoate (77.63 g, 397.98 mmol, 56.95 mL, 2.1 eq) was added over a period of 0.5 h. The mixture was stirred at 20 °C for 1 h. The reaction mixture was quenched by addition H₂O (300 mL) at 0 °C, and then extracted with PE / EtOAc = 10 / 1 (300 mL × 2). The combined organic layers were washed with brine (500 mL), dried over anhydrous sodium sulfate, filtered and concentrated under reduced pressure to give a residue. The crude product was used into the next step without further purification and dimethyl 6-isocyano-6-(p-tolylsulfonyl)undecanedioate (19d) (80 g, crude) was obtained as a black oil. [00417] Step 5: dimethyl 6-oxoundecanedioate (19e)

[00418] To a 1000 mL three neck round bottom flask was added DCM (500 mL) , followed by the addition of dimethyl 6-isocyano-6-(p-tolylsulfonyl)undecanedioate (19d) (80 g, 188.89 mmol, 1.0 eq) under N2, HCl (12 M, 112.71 mL, 7.2 eq) was added in portions over a period of 1 h. The two-phase mixture was stirred for 12 h at 25 °C. The reaction mixture was quenched by addition H₂O (150 mL) at 0 °C, and then extracted with DCM (150 mL × 2). The combined organic layers were washed with brine (200 mL), dried over anhydrous sodium sulfate, filtered and concentrated under reduced pressure to give a residue. The residue was purified by column chromatography (SiO₂, PE / EtOAc = 1 / 0 to 10 / 1) to give dimethyl 6-oxoundecanedioate (19e) (27 g, 104.53 mmol, 60% yield) as a colorless oil. ¹H NMR (400 MHz, CDCl₃) δ = 3.64 (s, 6H), 2.44 - 2.36 (m, 4H), 2.32 - 2.25 (m, 4H), 1.65 - 1.51 (m, 8H). [00419] Step 6: 5-(((1-methylpiperidin-4-yl)((octylthio)carbonyl)amino)methyl)isophthalic acid (19f):

[00420] A mixture of (1-methyl-4-piperidyl)methanamine (4.5 g, 35.10 mmol, 1.0 eq), NaBH(OAc)₃ (14.88 g, 70.19 mmol, 2.0 eq), HOAc (4.22 g, 70.19 mmol, 4.02 mL, 2.0 eq) in DCM (150 mL) was degassed and purged with N23 times, dimethyl 6-oxoundecanedioate (19e) (10.88 g, 42.12 mmol, 1.2 eq) in DCM (15 mL) was added and then the mixture was stirred at 20 °C for 10 h under N₂. The mixture pH was adjusted to 9 - 10 was with 1 M NaOH (30 mL), the aqueous phase was extracted with EtOAc (35 mL × 3). The combined organic phase was washed with brine (40 mL), dried with anhydrous sodium sulfate, filtered and concentrated in vacuum to give crude product. The residue was purified by flash silica gel chromatography (ISCO®; 120 g SepaFlash® Silica Flash Column, Eluent of 0 ~ 12% MeOH / DCM gradient @100 mL / min) to give dimethyl 6-[(1-methyl-4-piperidyl)methylamino]undecanedioate (19f) (8.34 g, 21.38 mmol, 61% yield, 95% purity) as a yellow oil. ¹H NMR (400 MHz, CDCl₃) δ = 3.65 (s, 6H), 2.83 (d, J = 11.6 Hz, 2H), 2.47 - 2.37 (m, 3H), 2.30 (t, J = 7.6 Hz, 4H), 2.24 (s, 3H), 1.93 - 1.84 (m, 2H), 1.75 - 1.66 (m, 2H), 1.65 - 1.56 (m, 4H), 1.39 - 1.17 (m, 12H). [00421] Step 7: dimethyl 6-((chlorocarbonyl)((1-methylpiperidin-4- yl)methyl)amino)undecanedioate (19g)

[00422] To a solution of dimethyl 6-[(1-methyl-4-piperidyl)methylamino]undecanedioate (19f) (2 g, 5.40 mmol, 1.0 eq) and TEA (1.64 g, 16.19 mmol, 2.25 mL, 3.0 eq) dissolved in dry DCM (20 mL) was added triphosgene (961.06 mg, 3.24 mmol, 0.6 eq) at 0 °C under N₂. The resulting solution was stirred at 25 °C for 1 h (checked by TLC) and concentrated under reduced pressure and kept under N2. The reaction mixture was concentrated in vacuum to give residue to give dimethyl 6-[chlorocarbonyl-[(1-methyl-4-piperidyl)methyl]amino]undecanedioate (19g) (2.34 g, 5.40 mmol, 100% yield) as a yellow solid. [00423] Step 8: dimethyl 6-(((heptylthio)carbonyl)((1-methylpiperidin-4- yl)methyl)amino)undecanedioate (19h)

[00424] To a solution of heptane-1-thiol (2.14 g, 16.21 mmol, 2.54 mL, 3.0 eq) dissolved in dry THF (30 mL) was added NaOH (648.48 mg, 16.21 mmol, 3.0 eq) at 0 °C under N2. Then dimethyl 6-[chlorocarbonyl-[(1-methyl-4-piperidyl)methyl]amino]undecanedioate (19g) (2.34 g, 5.40 mmol, 1.0 eq) dissolved in THF (10 mL) was added via syringe slowly under N2 at 0 °C. The resulting solution was stirred at 25 ° C for 15 h. The reaction mixture was quenched by NH₄Cl (10 mL) at 0 °C and then diluted with EtOAc (5 mL). The aqueous phase was extracted with EtOAc (20 mL × 2). The combined organic phase was washed with brine (30 mL), dried over with anhydrous sodium sulfate, filtered and concentrated in vacuum to give residue. The residue was purified by flash silica gel chromatography (ISCO®; 40 g SepaFlash® Silica Flash Column, Eluent of 0 ~ 6% MeOH / DCM gradient @ 60 mL / min) to give dimethyl 6- [heptylsulfanylcarbonyl-[(1-methyl-4-piperidyl)methyl]amino]undecanedioate (19h) (1.99 g, 3.58 mmol, 66.2% yield, 95% purity) as a yellow oil. ¹H NMR (400 MHz, CDCl3) δ = 3.89 - 3.75 (m, 1H), 3.66 (s, 6H), 3.15 - 2.98 (m, 2H), 2.94 - 2.80 (m, 4H), 2.39 - 2.24 (m, 7H), 2.06 - 1.86 (m, 2H), 1.83 - 1.73 (m, 1H), 1.72 - 1.52 (m, 10H), 1.47 - 1.22 (m, 16H), 0.94 - 0.81 (m, 3H). [00425] Step 9: 6-(((heptylthio)carbonyl)((1-methylpiperidin-4- yl)methyl)amino)undecanedioic acid (19i)

[00426] A mixture of dimethyl 6-[heptylsulfanylcarbonyl-[(1-methyl-4- piperidyl)methyl]amino]undecanedioate (19h) (1.99 g, 3.76 mmol, 1.0 eq), LiOH•H2O (473.77 mg, 11.29 mmol, 3.0 eq) in THF (18 mL) and H₂O (6 mL) was degassed and purged with N₂3 times, and then the mixture was stirred at 25 °C for 2 h under N₂. The reaction mixture was adjust pH = 3 - 4 with 1 M HCl, and then diluted with H2O (20 mL) and extracted with EtOAc 60 mL (20 mL × 3). The combined organic layers were washed with brine (15 mL), dried over anhydrous sodium sulfate, filtered and concentrated under reduced pressure to give 6- [heptylsulfanylcarbonyl-[(1-methyl-4-piperidyl)methyl]amino]undecanedioic acid (19i) (1.93 g, crude) as a white solid.¹H NMR (400 MHz, DMSO-d6) δ = 3.82 - 3.67 (m, 1H), 3.35 - 3.27 (m, 2H), 3.15 - 3.03 (m, 2H), 2.97 - 2.82 (m, 2H), 2.79 (t, J = 7.2 Hz, 2H), 2.66 (s, 3H), 2.19 (t, J = 7.2 Hz, 4H), 1.82 - 1.71 (m, 3H), 1.58 - 1.43 (m, 12H), 1.29 - 1.15 (m, 12H), 0.85 (t, J = 6.8 Hz, 3H). [00427] Step 10: bis(6,6-bis(pentyloxy)hexyl) 6-(((heptylthio)carbonyl)((1-methylpiperidin-4- yl)methyl)amino)undecanedioate (19)

[00428] To a solution of 6-[heptylsulfanylcarbonyl-[(1-methyl-4- piperidyl)methyl]amino]undecanedioic acid (19i) (500 mg, 998.54 μmol, 1.0 eq), 6,6- dipentoxyhexan-1-ol (19c) (822.11 mg, 3.00 mmol, 3.0 eq) and DMAP (365.96 mg, 3.00 mmol, 3.0 eq) in DCM (13 mL) was added EDCI (574.26 mg, 3.00 mmol, 3.0 eq) . The mixture was stirred at 30 °C for 16 h. The reaction mixture was partitioned between DCM (10 mL) and H₂O (20 mL). The organic phase was separated, washed with brine 20 mL (10 mL × 2), dried over anhydrous sodium sulfate, filtered and concentrated under reduced pressure to give a residue. The residue was purified by flash silica gel chromatography (ISCO®; 20 g SepaFlash® Silica Flash Column, Eluent of 0 ~ 6% MeOH / DCM gradient @ 45 mL / min). then was further purified by prep-HPLC (FA condition; column: CD11-Welch Xtimate C1150 * 25 * 10 μm; mobile phase: [water (FA) - MeOH]; gradient: 63% - 93% B over 8 min) to give bis(6,6- dipentoxyhexyl) 6-[heptylsulfanylcarbonyl-[(1-methyl-4- piperidyl)methyl]amino]undecanedioate (19) (300 mg, 295.95 μmol, 37.50% yield, 99.99% purity) as a yellow oil. LCMS: [M+H]⁺: 1013.4¹H NMR (400 MHz, CDCl₃) δ = 4.46 (t, J = 5.6 Hz, 2H), 4.05 (t, J = 6.8 Hz, 4H), 3.82 (s, 1H), 3.63-3.51 (m, 4H), 3.45 - 3.35 (m, 4H), 3.17 - 2.98 (m, 2H), 2.95 - 2.81 (m, 4H), 2.34 - 2.23 (m, 7H), 2.03- 1.87 (m, 2H), 1.70 - 1.53 (m, 27H), 1.50 - 1.44 (m, 2H), 1.41 - 1.26 (m, 38H), 0.96 - 0.84 (m, 15H). Example 20: Synthesis of [11-(6,6-dipentoxyhexanoyloxy)-6-[heptylsulfanylcarbonyl-[(1- methyl-4-piperidyl)methyl]amino]undecyl] 6,6-dipentoxyhexanoate (20) [00429] Step 1: dimethyl 6-[benzyloxycarbonyl-[(1-methyl-4- piperidyl)methyl]amino]undecanedioate (20a):

[00430] To a solution of dimethyl 6-[(1-methyl-4-piperidyl)methylamino]undecanedioate (19f) (3.5 g, 9.45 mmol, 1 eq) and DIPEA (1.46 g, 11.34 mmol, 1.97 mL, 1.2 eq) in DCM (50 mL) was added CbzOSu (2.82 g, 11.34 mmol, 1.2 eq) at 0 °C under N2. After addition, the mixture was stirred at 20 °C for 16 h. After addition, the mixture was stirred at 20 °C for 3 h. The reaction mixture was concentrated under reduced pressure to give residue. The residue was purified by flash silica gel chromatography (40 g SepaFlash® Silica Flash Column, DCM : MeOH: 0~10%) to give dimethyl 6-[benzyloxycarbonyl-[(1-methyl-4- piperidyl)methyl]amino]undecanedioate (20a) (3.8 g, 7.53 mmol, 80.9% yield) as yellow oil. LCMS: [M+H⁺]:505.3 [00431] Step 2: benzyl N-[6-hydroxy-1-(5-hydroxypentyl)hexyl]-N-[(1-methyl-4- piperidyl)methyl]carbamate (20b)

[00432] To a solution of dimethyl 6-[benzyloxycarbonyl-[(1-methyl-4- piperidyl)methyl]amino]undecanedioate (20a) (3.5 g, 6.94 mmol, 1 eq) in THF (60 mL) was added dropwise LAH (2.5 M, 6.94 mL, 2.5 eq) at 0 °C under N2. After addition, the mixture was stirred at 25 °C for 16 h. The reaction mixture was quenched by addition H2O (0.7 mL) at 0 °C under N₂, and then successively added 15% aq. NaOH (0.7 mL), H₂O (2.1 mL). The reaction mixture was diluted with THF (50 mL) and added N2SO4. The reaction mixture was filtered and the filter cake was washed with THF (50 mL × 3). The filtrate concentrated under reduced pressure to give benzyl N-[6-hydroxy-1-(5-hydroxypentyl)hexyl]-N-[(1-methyl-4- piperidyl)methyl]carbamate (20b) (2.3 g, 4.86 mmol, 70.1% yield, 94.8% purity) as light yellow oil. The crude product was used for next step further purification. LCMS: [M+H⁺]:449.3 [00433] Step 3: [6-[benzyloxycarbonyl-[(1-methyl-4-piperidyl)methyl]amino]-11-(6,6- dipentoxyhexanoyloxy)undecyl] 6,6-dipentoxyhexanoate (20c)

[00434] To a solution of benzyl N-[6-hydroxy-1-(5-hydroxypentyl)hexyl]-N-[(1-methyl-4- piperidyl)methyl]carbamate (20b, 1.3 g, 2.90 mmol, 1 eq) and 6,6-dipentoxyhexanoic acid (2.09 g, 7.24 mmol, 2.5 eq) in DCM (30 mL) were added EDCI (1.67 g, 8.69 mmol, 3 eq) and DMAP (1.06 g, 8.69 mmol, 3 eq). The mixture was stirred at 20 °C for 16h. The reaction mixture was quenched with H₂O (50 mL) and then diluted with DCM (30 mL). The aqueous phase was extracted with DCM (40 mL × 3). The combined organic phases were washed with brine (50 mL), dried with anhydrous Na2SO4, filtered and concentrated in vacuum to give residue. The residue was purified by flash silica gel chromatography (40 g SepaFlash® Silica Flash Column, DCM : MeOH: 0~10%) to give [6-[benzyloxycarbonyl-[(1-methyl-4-piperidyl)methyl]amino]- 11-(6,6-dipentoxyhexanoyloxy)undecyl] 6,6-dipentoxyhexanoate (20c, 2.3 g, 2.32 mmol, 82.1% yield) as yellow oil. [00435] Step 4: [11-(6,6-dipentoxyhexanoyloxy)-6-[(1-methyl-4- piperidyl)methylamino]undecyl] 6,6-dipentoxyhexanoate (20d)

[00436] To a solution of [6-[benzyloxycarbonyl-[(1-methyl-4-piperidyl)methyl]amino]-11- (6,6-dipentoxyhexanoyloxy)undecyl] 6,6-dipentoxyhexanoate (20c, 2.3 g, 2.32 mmol, 1 eq) in MeOH (20 mL) and THF (20 mL) were added Pd/C (200 mg, 187.93 μmol, 10% purity) and Pd(OH)₂/C (200 mg, 284.83 μmol, 20% purity) under N₂. The suspension was degassed under vacuum and purged with H2 several times. The mixture was stirred under H2 (15 psi) at 40 °C for 3 h. The reaction mixture was filtered through a pad of diatomaceous earth and the filter cake was washed with THF (20 mL × 3), then the filtrate was concentrated under reduced pressure to give residue. The residue was purified by flash silica gel chromatography (20 g SepaFlash® Silica Flash Column, DCM: MeOH: 0~10%) to give 1[11-(6,6-dipentoxyhexanoyloxy)-6-[(1- methyl-4-piperidyl)methylamino]undecyl] 6,6-dipentoxyhexanoate (20d, 1.62 g, 1.89 mmol, 81.0% yield) as colorless oil. ¹H NMR (400 MHz, CDCl₃) δ = 4.46 (t, J = 5.6 Hz, 2H), 4.06 (t, J = 6.8 Hz, 4H), 3.63 - 3.52 (m, 4H), 3.45 - 3.34 (m, 4H), 2.96 - 2.90 (m, 2H), 2.50 - 2.43 (m, 2H), 2.38 - 2.27 (m, 7H), 2.04 - 1.94 (m, 2H), 1.80 - 1.73 (m, 3H), 1.68 - 1.54 (m, 20H), 1.46 - 1.22 (m, 35H), 0.91 (t, J = 6.4 Hz, 12H). [00437] Step 5: [6-[chlorocarbonyl-[(1-methyl-4-piperidyl)methyl]amino]-11-(6,6- dipentoxyhexanoyloxy)undecyl] 6,6-dipentoxyhexanoate (20e)

[00438] To a solution of [11-(6,6-dipentoxyhexanoyloxy)-6-[(1-methyl-4- piperidyl)methylamino]undecyl] 6,6-dipentoxyhexanoate (20d, 800 mg, 935.32 μmol, 1 eq) and TEA (189.29 mg, 1.87 mmol, 260.37 μL, 2 eq) in DCM (15 mL) was added triphosgene (166.53 mg, 561.19 μmol, 0.6 eq) at 0 °C under N₂. After addition, the mixture was stirred at 20°C for 2 h. The reaction mixture was concentrated under reduced pressure to give [6-[chlorocarbonyl-[(1- methyl-4-piperidyl)methyl]amino]-11-(6,6-dipentoxyhexanoyloxy)undecyl] 6,6- dipentoxyhexanoate (20e, 858 mg, crude) as yellow solid. The crude product was used for next step starting material. [00439] Step 6: [11-(6,6-dipentoxyhexanoyloxy)-6-[heptylsulfanylcarbonyl-[(1-methyl-4- piperidyl)methyl]amino]undecyl] 6,6-dipentoxyhexanoate (20)

[00440] To a solution of heptane-1-thiol (306.25 mg, 2.32 mmol, 362.85 μL, 2.5 eq) dissolved in dry THF (10 mL) was added NaOH (92.62 mg, 2.32 mmol, 2.5 eq) at 0 °C under N2. After addition, the resulting solution was stirred at 0 °C for 0.5 h, and then [6-[chlorocarbonyl-[(1- methyl-4-piperidyl)methyl]amino]-11-(6,6-dipentoxyhexanoyloxy)undecyl] 6,6- dipentoxyhexanoate (20e, 850 mg, 926.15 μmol, 1 eq) in THF (10 mL)was added dropwise. The resulting solution was stirred at 20 °C for 15.5 h. The reaction mixture was quenched by NH₄Cl (50 mL) at 0 °C and then diluted with DCM (30 mL). The aqueous phase was extracted with DCM (30 mL × 3). The combined organic phase was washed with brine (60 mL), dried with anhydrous Na₂SO₄, filtered and concentrated in vacuum to give residue. The residue was purified by flash silica gel chromatography (12 g SepaFlash® Silica Flash Column, DCM : MeOH: 0~7%) and purified by prep-HPLC (column: CD11-Welch Xtimate C1150 × 25 × 10um; mobile phase: [water (FA) - MeOH]; gradient: 63% - 93% B over 8 min) to give [11- (6,6-dipentoxyhexanoyloxy)-6-[heptylsulfanylcarbonyl-[(1-methyl-4- piperidyl)methyl]amino]undecyl] 6,6-dipentoxyhexanoate (20, 245 mg, 238.99 μmol, 40.1% yield, 98.87% purity) as yellow oil. LCMS: [M+H⁺]:1014.0¹H NMR (400 MHz, CDCl₃) δ = 4.46 (t, J = 5.6 Hz, 2H), 4.05 (t, J = 6.8 Hz, 4H), 3.86 - 3.80 (m, 1H), 3.60 - 3.52 (m, 4H), 3.44 - 3.36 (m, 4H), 3.12 - 3.02 (m, 2H), 2.95 - 2.84 (m, 4H), 2.35 - 2.28 (m, 7H), 1.96 - 1.91 (m, 2H), 1.73 - 1.56 (m, 25H), 1.44 - 1.26 (m, 42H), 0.93 - 0.86 (m, 15H). Example 21: Synthesis of bis(2-butyloctyl) 10-[(1-methyl-4-piperidyl)-octylsulfinyl- amino]nonadecanedioate (21) [00441] Step 1: bis(2-butyloctyl) 10-[(1-methyl-4-piperidyl)-octylsulfinyl- amino]nonadecanedioate (21)

[00442] To a solution of bis(2-butyloctyl) 10-[(1-methyl-4-piperidyl)amino]nonadecanedioate (2b, 700 mg, 900.56 μmol, 1 eq) and TEA (273.38 mg, 2.70 mmol, 376.04 μL, 3 eq) in DCM (15 mL) was added dropwise octane-1-sulfinyl chloride (7b, 531.52 mg, 2.70 mmol, 3 eq) in DCM (5 mL) at 0 °C under N₂. The resulting mixture was stirred at 20 °C for 16 h. The reaction mixture was concentrated under reduced pressure to residue. The residue was purified by flash silica gel chromatography (20 g SepaFlash® Silica Flash Column, DCM : MeOH: 0~10%) to afford (21) (465 mg, 495.91 μmol, 55.0% yield, 99.99% purity) as yellow oil. LCMS: [M+H]⁺: 938.1¹H NMR (400 MHz, CDCl3) δ = 3.98 (d, J = 5.6 Hz, 4H), 3.75 - 3.60 (m, 1H), 3.45 - 3.35 (m, 2H), 3.20 - 3.15 (m, 1H), 2.84 - 2.71 (m, 2H), 2.66 (s, 3H), 2.63 - 2.51 (m, 2H), 2.30 (t, J = 7.6 Hz, 4H), 2.10 - 1.98 (m, 2H), 1.78 - 1.68 (m, 4H), 1.67 - 1.60 (m, 8H), 1.46 - 1.40 (m, 4H), 1.37 - 1.26 (m, 60H), 0.99 - 0.87 (m, 15H). Example 22: Synthesis of bis(2-butyloctyl) 10-[3-(dimethylamino)propyl- (octylsulfonimidoyl)amino]nonadecanedioate (22) [00443] Step 1: bis(2-butyloctyl) 10-((3- (dimethylamino)propyl)(octylsulfinyl)amino)nonadecanedioate (22a)

[00444] To a solution of bis(2-butyloctyl) 10-[3- (dimethylamino)propylamino]nonadecanedioate (1h) (700 mg, 914.69 μmol, 1.0 eq), DMAP (22.35 mg, 182.94 μmol, 0.2 eq) and TEA (277.67 mg, 2.74 mmol, 381.94 μL, 3.0 eq) in DCM (20 mL) was added octane-1-sulfinyl chloride (7b) (899.77 mg, 4.57 mmol, 5.0 eq) in DCM (1 mL) at 0 °C. The mixture was stirred at 25 °C for 1 h. The reaction was concentrated. The residue was purified by flash silica gel chromatography (ISCO®; 20g SepaFlash® Silica Flash Column, Eluent of 0 ~ 5% MeOH / DCM gradient @ 20 mL / min) to give bis(2-butyloctyl) 10- [3-(dimethylamino)propyl-octylsulfinyl-amino]nonadecanedioate (22a) (411 mg, 421.85 μmol, 46.1% yield, 95.0% purity) as a yellow oil. ¹H NMR (400 MHz, CDCl3) δ = 3.97 (d, J = 5.6 Hz, 4H), 3.24 - 3.21 (m, 1H), 3.18 - 2.94 (m, 4H), 2.92 - 2.70 (m, 6H), 2.30 (t, J = 7.6 Hz, 4H), 2.12 - 2.01 (m, 2H), 1.70 - 1.54 (m, 10H), 1.45 - 1.40 (m, 4H), 1.37 - 1.19 (m, 62H), 0.94 - 0.83 (m, 15H). [00445] Step 2: bis(2-butyloctyl) 10-[3-(dimethylamino)propyl- (octylsulfonimidoyl)amino]nonadecanedioate (22)

[00446] A mixture of bis(2-butyloctyl) 10-[3-(dimethylamino)propyl-octylsulfinyl- amino]nonadecanedioate (22a) (200 mg, 216.09 μmol, 1.0 eq), PhI(OAc)2 (208.80 mg, 648.26 μmol, 3.0 eq) and ammonium carbamate (67.48 mg, 864.34 μmol, 4.0 eq) in MeOH (2 mL) was degassed and purged with N₂3 times, and then the mixture was stirred at 20 °C for 2 h under N₂. The reaction was concentrated. The residue was purified by prep-HPLC (FA condition; column: CD11-Welch Xtimate C1150 × 25 × 10 μm; mobile phase: [water (FA) - MeOH]; gradient: 70% - 100% B over 8 min) to give bis(2-butyloctyl)10-[3-(dimethylamino)propyl- (octylsulfonimidoyl)amino]nonadecanedioate (22) (150 mg, 157.48 μmol, 72.9% yield, 98.75% purity) as a yellow oil. LCMS: [M+H]⁺: 940.9¹H NMR (400 MHz, CDCl3) δ = 6.90 (s, 1H), 3.96 (d, J = 5.6 Hz, 4H), 3.82 - 3.60 (m, 2H), 3.58 - 3.34 (m, 6H), 3.32 - 3.12 (m, 2H), 3.08 - 2.95 (m, 1H), 2.90 - 2.76 (m, 1H), 2.71 - 2.56 (m, 1H), 2.30 (t, J = 7.6 Hz, 4H), 2.22 - 2.08 (m, 4H), 1.65 - 1.58 (m, 8H), 1.47 - 1.40 (m, 4H), 1.32 - 1.23 (m, 60H), 0.91 - 0.86 (m, 15H). Example 23: Synthesis of bis(2-butyloctyl) 10-[(1-methyl-4-piperidyl)methyl- (octylsulfonimidoyl)amino]nonadecanedioate (23)

[00447] A mixture of bis(2-butyloctyl)10-[(1-methyl-4-piperidyl)methyl-octylsulfinyl- amino]nonadecanedioate (12) (200 mg, 210.17 μmol, 1.0 eq), PhI(OAc)₂ (203.09 mg, 630.52 μmol, 3.0 eq) and ammonium carbamate (65.63 mg, 840.69 μmol, 4.0 eq) in MeOH (5 mL) was degassed and purged with N23 times, and then the mixture was stirred at 20 °C for 2 h under N2. The reaction was concentrated. The residue was purified by prep-HPLC (FA condition; column: CD12-ACCHROM phenyl - Hexyl 150 × 25 × 10 μm; mobile phase: [water (FA) - ACN]; gradient: 60% - 90% B over 10 min) to give bis(2-butyloctyl) 10-[(1-methyl-4-piperidyl)methyl- (octylsulfonimidoyl)amino]nonadecanedioate (23) (105.39 mg, 104.06 μmol, 49.51% yield, 99.99% purity, FA salt) as a colorless oil. LCMS: [M+H]⁺: 966.9¹H NMR (400 MHz, CDCl₃) δ = 8.74 (s, 1H), 4.20 - 4.10 (m, 1H), 3.96 (d, J = 5.2 Hz, 4H), 3.80 - 3.48 (m, 3H), 3.18 - 2.90 (m, 4H), 2.88 - 2.68 (m, 2H), 2.59 - 2.42 (m, 1H), 2.35 - 2.23 (m, 4H), 2.20 - 1.95 (m, 2H), 1.89 - 1.72 (m, 3H), 1.71 - 1.54 (m, 10H), 1.52 - 1.38 (m, 6H), 1.37 - 1.17 (m, 60H), 1.01 - 0.77 (m, 15H). Example 24: Synthesis of bis(2-butyloctyl) 10-[(1-methyl-4-piperidyl)methyl-(2- propanoyloxyheptylsulfanylcarbonyl)amino]nonadecanedioate (24) [00448] Step 1: 1-tritylsulfanylheptan-2-ol (24a)

NaOH (1.2 eq) M_{eCN, 50 °C, 3 h} 24a [00449] To a solution of triphenylmethanethiol (7.26 g, 26.27 mmol, 1.2 eq) in MeCN (60 mL) were added NaOH (1.05 g, 26.27 mmol, 1.2 eq) and 2-pentyloxirane (2.5 g, 21.89 mmol, 1 eq). The mixture was stirred at 50 °C for 3 h. The reaction mixture was filter and the filtrate was concentrated under reduced pressure to give residue. The residue was purified by flash silica gel chromatography (120 g SepaFlash® Silica Flash Column, PE : EtOAc: 0~10%) to give 1- tritylsulfanylheptan-2-ol (24a, 7.9 g, 20.23 mmol, 92.4% yield) as yellow oil. [00450] Step 2: 1-(tritylsulfanylmethyl)hexyl propanoate (24b)

[00451] To a solution of 1-tritylsulfanylheptan-2-ol (24a, 4 g, 10.24 mmol, 8.33e-1 eq) and pyridine (1.94 g, 24.58 mmol, 1.98 mL, 2 eq), DMAP (150.14 mg, 1.23 mmol, 0.1 eq) in DCM (50 mL) was added dropwise propionyl chloride (1.36 g, 14.75 mmol, 1.36 mL, 1.2 eq) at 0 °C under N2. After addition, the mixture was stirred at 20 °C for 3 h. The reaction mixture was concentrated under reduced pressure to give residue. The residue was purified by flash silica gel chromatography (80 g SepaFlash® Silica Flash Column, PE : EtOAc: 0~10%) to give 1- (tritylsulfanylmethyl)hexyl propanoate (24b, 4.1 g, 9.18 mmol, 74.7% yield) as yellow oil. [00452] Step 3: 1-(sulfanylmethyl)hexyl propanoate (24c)

[00453] To a solution of 1-(tritylsulfanylmethyl)hexyl propanoate (24b, 3 g, 6.72 mmol, 1 eq) and triisopropylsilane (2.13 g, 13.43 mmol, 2.76 mL, 2 eq) in DCM (40 mL) was added dropwise TFA (10.23 g, 89.75 mmol, 6.67 mL, 13.36 eq) at 0 °C under N2. After addition, the mixture was stirred at 20 °C for 3 hr. The reaction mixture was concentrated under reduced pressure to remove TFA and filtered. The filtrate was concentrated under reduced pressure to give residue. The residue was distilled in vacuum (oil pump pressure: 0.96 bar; oil bath temperature: 140 °C; Steam temperature: 78~80°C) to give 1-(sulfanylmethyl)hexyl propanoate (24c, 410 mg, 2.01 mmol, 29.9% yield) as colorless liquid. ¹H NMR (400 MHz, CDCl₃) δ = 4.94 - 4.88 (m, 1H), 2.74 - 2.65 (m, 2H), 2.36 (q, J = 7.6 Hz, 2H), 1.70 - 1.63 (m, 2H), 1.34 - 1.29 (m, 6H), 1.17 (t, J = 7.6 Hz, 3H), 0.92 - 0.89 (m, 3H). [00454] Step 4: bis(2-butyloctyl) 10-[chlorocarbonyl-[(1-methyl-4- piperidyl)methyl]amino]nonadecanedioate (24d)

[00455] To a solution of bis(2-butyloctyl) 10-[(1-methyl-4- piperidyl)methylamino]nonadecanedioate (5f, 500 mg, 631.86 μmol, 1 eq) and TEA (127.87 mg, 1.26 mmol, 175.89 μL, 2 eq) dissolved in dry DCM (15 mL) was added triphosgene (112.50 mg, 379.11 μmol, 0.6 eq) at 0 °C under N2. The resulting solution was stirred at 20 °C for 2 h. The reaction mixture was concentrated under reduced pressure to give bis(2-butyloctyl) 10- [chlorocarbonyl-[(1-methyl-4-piperidyl)methyl]amino]nonadecanedioate (24d, 540 mg, crude) as a yellow solid. The crude product was used for next step starting material. [00456] Step 5: bis(2-butyloctyl) 10-[(1-methyl-4-piperidyl)methyl-(2- propanoyloxyheptylsulfanylcarbonyl)amino]nonadecanedioate (24)

[00457] To a solution of 1-(sulfanylmethyl)hexyl propanoate (24c, 143.59 mg, 702.76 μmol, 1.5 eq) dissolved in dry THF (3 mL) was added dropwise bis(2-butyloctyl) 10-[chlorocarbonyl- [(1-methyl-4-piperidyl)methyl]amino]nonadecanedioate (24d, 400 mg, 468.51 μmol, 1 eq) in THF (5 mL) at 0 °C under N₂. After addition, the resulting solution was stirred at 20 °C for 3 h. The reaction mixture was concentrated under reduced pressure to give residue. The residue was purified by flash silica gel chromatography (12 g SepaFlash® Silica Flash Column, DCM : MeOH: 0~7%) and prep-HPLC (column: CD12-ACCHROM phenyl-Hexyl 150 × 25 × 10um;mobile phase: [water (FA) - ACN]; gradient: 65% - 95% B over 10 min) to give 3- pentyloctyl 9-[decyl-[(1-methyl-4-piperidyl)methylsulfanylcarbonyl]amino]nonanoate (24, 760 mg, 1.12 mmol, 37.5% yield, 98.58% purity) as yellow oil. LCMS: [M+H⁺]: 1022.0¹H NMR (400 MHz, CDCl₃) δ = 5.08 - 5.00 (m, 1H), 4.02 - 3.95 (m, 4H), 3.80 - 3.65 (m, 1H), 3.58 - 3.48 (m, 2H), 3.30 - 3.21 (m, 1H), 3.14 - 3.06 (m, 1H), 3.00 - 2.85 (m, 2H), 2.78 - 2.72 (m, 3H), 2.70 - 2.65 (m, 2H), 2.34 - 2.27 (m, 7H), 1.95 - 1.85 (m, 4H), 1.80 - 1.73 (m, 3H), 1.64 - 1.61 (m, 4H), 1.50 - 1.40 (m, 4H), 1.35 - 1.23 (m, 58H), 1.14 (t, J = 7.2 Hz, 4H), 0.95 - 0.83 (m, 15H). Example 25: Synthesis of bis(2-butyloctyl) 10-[(N-ethyl-S-octyl-sulfonimidoyl)-[(1-methyl-4- piperidyl)methyl]amino]nonadecanedioate (25) [00458] Step 1: bis(2-butyloctyl) 10-[(1-methyl-4-piperidyl)methyl-octylsulfinyl- amino]nonadecanedioate (25a)

[00459] To a solution of bis(2-butyloctyl) 10-[(1-methyl-4- piperidyl)methylamino]nonadecanedioate (5f, 1.1 g, 1.39 mmol, 1 eq) and TEA (281.32 mg, 2.78 mmol, 386.96 μL, 2 eq) in DCM (15 mL) was added dropwise octane-1-sulfinyl chloride (7b, 546.96 mg, 2.78 mmol, 2 eq) in DCM (5 mL) at 0 °C under N2. After addition, the resulting mixture was stirred at 20 °C for 3 h. The reaction mixture was concentrated under reduced pressure to give residue. The residue was purified by flash silica gel chromatography (20 g SepaFlash® Silica Flash Column, DCM : MeOH: 0~7%) to give bis(2-butyloctyl) 10-[(1- methyl-4-piperidyl)methyl-octylsulfinyl-amino]nonadecanedioate (25a, 1.1 g, 1.16 mmol, 83.2% yield, 99.9% purity) as yellow oil. LCMS: [M+H] ⁺: 952.1 [00460] Step 2: bis(2-butyloctyl) 10-[(1-methyl-4-piperidyl)methyl- (octylsulfonimidoyl)amino]nonadecanedioate (25b)

[00461] A mixture of bis(2-butyloctyl) 10-[(1-methyl-4-piperidyl)methyl-octylsulfinyl- amino]nonadecanedioate (25b, 700 mg, 735.61 μmol, 1 eq), ammonium carbamate (229.72 mg, 2.94 mmol, 4 eq), PhI(OAc)2 (710.81 mg, 2.21 mmol, 3 eq) in MeOH (15 mL) was degassed and purged with N23 times, and then the mixture was stirred at 20 °C for 2 h under N₂ atmosphere. The reaction mixture was concentrated under reduced pressure to residue. The residue was purified by flash silica gel chromatography (12 g SepaFlash® Silica Flash Column, DCM : MeOH: 0~10%) to give bis(2-butyloctyl) 10-[(1-methyl-4-piperidyl)methyl- (octylsulfonimidoyl)amino]nonadecanedioate (25c, 610 mg, 629.81 μmol, 85.6% yield, 99.8% purity) as yellow oil. ¹H NMR (400 MHz, CDCl₃) δ = 4.19 (br d, J = 12.0 Hz, 2H), 3.98 (d, J = 6.0 Hz, 4H), 3.74 (s 3H), 3.20 - 3.05 (m, 3H), 2.99 - 2.76 (m, 3H), 2.58 - 2.50 (m, 1H), 2.31 (dt, J = 3.2, 7.2 Hz, 4H), 2.18 - 2.05 (m, 4H), 1.90 - 1.82 (m, 4H), 1.66 - 1.62 (m, 6H), 1.54 - 1.40 (m, 6H), 1.35 - 1.25 (m, 60H), 0.96 - 0.85 (m, 15H). [00462] Step 3: bis(2-butyloctyl) 10-[(N-ethyl-S-octyl-sulfonimidoyl)-[(1-methyl-4- piperidyl)methyl]amino]nonadecanedioate (25)

[00463] To a solution of bis(2-butyloctyl) 10-[(1-methyl-4-piperidyl)methyl- (octylsulfonimidoyl)amino]nonadecanedioate (25c, 500 mg, 517.27 μmol, 1 eq) in THF (10 mL) was added NaH (62.07 mg, 1.55 mmol, 60% purity, 3 eq) at 0 °C under N2. After addition, the mixture was stirred at 20 °C for 0.5 h, and then EtI (403.38 mg, 2.59 mmol, 206.86 μL, 5 eq) in THF (3 mL) was added dropwise at 20 °C. The resulting mixture was stirred at 20 °C for 23.5 h. The reaction mixture was quenched by NH4Cl (20 mL) at 0 °C and extracted with DCM (30 mL × 3). The combined organic phase was washed with brine (30 mL), dried with anhydrous Na₂SO₄, filtered and concentrated in vacuum to give residue. The residuewas purified by flash silica gel chromatography (12 g SepaFlash® Silica Flash Column, DCM : MeOH: 0~7%) and purified by prep-HPLC (column: CD12-ACCHROM phenyl-Hexyl 150 × 25 × 10um; mobile phase: [water (TFA) - (ACN-THF 2/1)]; gradient:54% - 84% B over 15 min) to give (25) (88 mg, 88.46 μmol, 59.5% yield, 99.99% purity) as yellow oil. LCMS: [M+H] ⁺: 995.2¹H NMR (400 MHz, CDCl3) δ = 4.25 (br d, J = 11.6 Hz, 1H), 4.09 (br d, J = 12.4 Hz, 1H), 3.96 (d, J = 5.6 Hz, 4H), 3.54 (s, 3H), 3.21 - 3.02 (m, 3H), 2.99 - 2.81 (m, 4H), 2.80 - 2.72 (m, 1H), 2.57 - 2.47 (m, 1H), 2.30 (dt, J = 3.2, 7.2 Hz, 4H), 2.02 - 1.89 (m, 3H), 1.85 -1.76 (m, 3H), 1.68 - 1.58 (m, 8H), 1.54 - 1.38 (m, 6H), 1.34 - 1.22 (m, 62H), 0.92 - 0.84 (m, 15H). Example 26: Synthesis of bis(2-butyloctyl) 10-[(1-methyl-4-piperidyl)methyl-nonanoyl- amino]nonadecanedioate (26)

[00464] To a solution of bis(2-butyloctyl) 10-[(1-methyl-4- piperidyl)methylamino]nonadecanedioate (5f, 500 mg, 631.86 μmol, 1 eq) and TEA (130.18 mg, 1.29 mmol, 179.07 μL, 2 eq) in DCM (10 mL) was added dropwise nonanoyl chloride (170.48 mg, 964.89 μmol, 181.36 μL, 1.5 eq) in DCM (3 mL) at 0 °C under N₂. The resulting mixture was stirred at 20 °C for 2 h. The reaction mixture was concentrated under reduced pressure to residue. The residue was purified by prep-HPLC (column: CD12-ACCHROM phenyl-Hexyl 150 × 25 × 10 μm; phase: [water (FA) - ACN]; gradient: 54% - 84% B over 10 min) to afford (26) (405 mg, 434.72 μmol, 67.5% yield, 99.99% purity) as yellow oil. LCMS: [M+H]⁺: 931.9 ¹H NMR (400 MHz, CDCl3) δ = 3.98 (d, J = 6.0 Hz, 4H), 3.68 - 3.53 (m, 1H), 3.05 (br d, J = 7.2 Hz, 1H), 3.00 - 2.96 (m, 1H), 2.94 - 2.85 (m, 2H), 2.35 - 2.23 (m, 9H), 1.99 - 1.82 (m, 2H), 1.75 - 1.60 (m, 11H), 1.47 - 1.40 (m, 4H), 1.29 (br s, 64H), 0.95 - 0.83 (m, 15H). Example 27: Synthesis of 6-(((heptylthio)carbonyl)((1-methylpiperidin-4- yl)methyl)amino)undecane-1,11-diyl bis(4,4-bis(hexyloxy)butanoate) (27) [00465] Step 1: 4,4-bis(hexyloxy)butanenitrile (27a):

[00466] A mixture of 4,4-dimethoxybutanenitrile (10 g, 77.43 mmol, 1.0 eq), PPTS (9.73 g, 38.71 mmol, 0.5 eq) and inhexan-1-ol (39.55 g, 387.13 mmol, 48.24 mL, 5.0 eq) was degassed and purged with N₂3 times, then the mixture was stirred at 110 °C for 16 h under N₂. The reaction was concentrated under reduced pressure to give a residue. The residue was purified by flash silica gel chromatography (ISCO®; 120 g SepaFlash® Silica Flash Column, Eluent of 0 ~ 3% EtOAc / PE gradient @ 100 mL / min) to give 4,4-dihexoxybutanenitrile (27a) (15.9 g, 59.02 mmol, 79.5% yield) as a yellow oil. ¹H NMR (400 MHz, CDCl₃) δ = 4.56 (t, J = 5.2 Hz, 1H), 3.66 - 3.54 (m, 2H), 3.49 - 3.37 (m, 2H), 2.42 (t, J = 7.2 Hz, 2H), 2.01 - 1.87 (m, 2H), 1.61 - 1.52 (m, 4H), 1.37 - 1.25 (m, 12H), 0.89 (t, J = 6.8 Hz, 6H). [00467] Step 2: 4,4-bis(hexyloxy)butanoic acid (27b)

[00468] A mixture of 4,4-dihexoxybutanenitrile (27a) (5 g, 18.56 mmol, 1.0 eq) and KOH (3.12 g, 55.67 mmol, 3.0 eq) in H2O (30 mL) and EtOH (30 mL) was degassed and purged with N23 times, and then the mixture was stirred at 110 °C for 16 h under N2. The reaction mixture was cooled down to 20 °C and concentrated to remove sovent, pH adjust to pH = 5 by addition 1N HCl, and extracted with EtOAc 200 mL (100 mL × 2). The combined organic layers were washed with brine (200 mL), dried over anhydrous sodium sulfate, filtered and concentrated under reduced pressure to give a residue. The residue was purified by flash silica gel chromatography (ISCO®; 80 g SepaFlash® Silica Flash Column, Eluent of 0 ~ 30% EtOAc / PE gradient @ 100mL / min) to give 4,4-dihexoxybutanoic acid (27b) (3.92 g, 13.59 mmol, 73.2% yield) as a yellow oil. ¹H NMR (400 MHz, CDCl3) δ = 4.45 (t, J = 5.6 Hz, 1H), 3.55 - 3.47 (m, 2H), 3.39-3.29 (m, 2H), 2.38 (t, J = 7.2 Hz, 2H), 1.92 - 1.83 (m, 2H), 1.55 - 1.44 (m, 4H), 1.30 - 1.18 (m, 12H), 0.82 (t, J = 6.8 Hz, 6H). [00469] Step 3: dimethyl 6-(((1-methylpiperidin-4-yl)methyl)amino)undecanedioate (27c)

[00470] A mixture of (1-methyl-4-piperidyl)methanamine (2.5 g, 19.50 mmol, 1.0 eq), NaBH(OAc)₃ (8.27 g, 39.00 mmol, 2.0 eq) and HOAc (2.34 g, 39.00 mmol, 2.23 mL, 2.0 eq) in DCM (45 mL) was degassed and purged with N23 times, then dimethyl 6-oxoundecanedioate (6.04 g, 23.40 mmol, 1.2 eq) in DCM (15 mL) was added and then the mixture was stirred at 20 °C for 10 h under N2. The mixture pH was adjusted to 9 - 10 was with 1 N NaOH (~30 mL), the aqueous phase was extracted with DCM 60 mL (20 mL × 3). The combined organic phase was washed with brine 80 mL (40 mL×2), dried with anhydrous sodium sulfate, filtered and concentrated in vacuum to give residue. The residue was purified by flash silica gel chromatography (ISCO®; 120 g SepaFlash® Silica Flash Column, Eluent of 0 ~ 14% MeOH / DCM gradient @100 mL / min) to give dimethyl 6-[(1-methyl-4- piperidyl)methylamino]undecanedioate (27c) (5.67 g, 14.54 mmol, 74.6% yield, 95.0% purity) as a yellow oil. ¹H NMR (400 MHz, CDCl3) δ = 3.63 (s, 6H), 2.90 - 2.79 (m, 2H), 2.44 - 2.36 (m, 3H), 2.31 - 2.26 (m, 4H), 2.24 (s, 3H), 1.95 - 1.83 (m, 2H), 1.74 - 1.66 (m, 2H), 1.64 - 1.54 (m, 4H), 1.40 - 1.14 (m, 12H). [00471] Step 4: dimethyl 6-(((benzyloxy)carbonyl)((1-methylpiperidin-4- yl)methyl)amino)undecanedioate (27d)

[00472] A mixture of dimethyl 6-[(1-methyl-4-piperidyl)methylamino]undecanedioate (27c) (5.17 g, 13.95 mmol, 1.0 eq) and DIEA (3.61 g, 27.91 mmol, 4.86 mL, 2.0 eq) in DCM (90 mL) was degassed and purged with N23 times, then benzyl (2,5-dioxopyrrolidin-1-yl) carbonate (4.17 g, 16.74 mmol, 1.2 eq) was added at 0 °C in portions, and the reaction was stirred at 25 °C for 3 h. The reaction mixture was concentrated under reduced pressure to give a residue. The residue was purified by flash silica gel chromatography (ISCO®; 80 g SepaFlash® Silica Flash Column, Eluent of 0 ~ 5% MeOH / DCM gradient @ 100 mL / min) to give dimethyl 6- [benzyloxycarbonyl-[(1-methyl-4-piperidyl)methyl]amino]undecanedioate (27d) (3.47 g, 6.19 mmol, 44.4% yield, 90% purity) as a yellow oil. ¹H NMR (400 MHz, CDCl₃) δ =7.43 - 7.27 (m, 5H), 5.10 (s, 2H), 3.83 - 3.68 (m, 1H), 3.64 (s, 6H), 3.33 - 3.11 (m, 2H), 3.00 (d, J = 6.0 Hz, 2H), 2.43 (s, 3H), 2.31 - 2.18 (m, 5H), 1.92 - 1.71 (m, 2H), 1.62 - 1.44 (m, 10H), 1.42 - 1.08 (m, 6H). [00473] Step 5: benzyl (1,11-dihydroxyundecan-6-yl)((1-methylpiperidin-4- yl)methyl)carbamate (27e)

[00474] A mixture of dimethyl 6-[benzyloxycarbonyl-[(1-methyl-4- piperidyl)methyl]amino]undecanedioate (27d) (3.27 g, 6.48 mmol, 1.0 eq) in THF (50 mL) was degassed and purged with N23 times and cooled down to 0 °C, then LiAlH4 (2.5 M, 5.44 mL, 2.1 eq) was added via injection at 0 °C, and then the mixture was stirred at 25 °C for 1hr under N₂. After completion, the reaction mixture was diluted with THF (20 mL), then successively was added H₂O (0.5 mL), aq. NaOH (0.5 mL, 15%), H₂O (1.5 mL) and dried over anhydrous sodium sulfate at 0 °C under N2. The reaction mixture was filtered and the filtrate was concentrated in vacuum to give residue. The residue was purified by flash silica gel chromatography (ISCO®; 80 g SepaFlash® Silica Flash Column, Eluent of 0 ~ 14% MeOH / DCM gradient @ 50 mL / min) to give benzyl N-[6-hydroxy-1-(5-hydroxypentyl)hexyl]-N-[(1-methyl-4- piperidyl)methyl]carbamate (27e) (3.01 g, 6.37 mmol, 49.2% yield, 95.0% purity) as a colorless oil. ¹H NMR (400 MHz, CDCl₃) δ = 7.41 - 7.23 (m, 5H), 5.12 (s, 2H), 3.97 - 3.69 (m, 1H), 3.68 - 3.47 (m, 4H), 3.06 - 2.93 (m, 2H), 2.90 - 2.75 (m, 2H), 2.30 - 2.18 (m, 3H), 1.97 - 1.73 (m, 4H), 1.70 - 1.65 (m, 1H), 1.64 - 1.42 (m, 10H), 1.39 - 1.11 (m, 10H). [00475] Step 6: 6-(((benzyloxy)carbonyl)((1-methylpiperidin-4-yl)methyl)amino)undecane- 1,11-diyl bis(4,4-bis(hexyloxy)butanoate) (27f)

[00476] To a solution of benzyl N-[6-hydroxy-1-(5-hydroxypentyl)hexyl]-N-[(1-methyl-4- piperidyl)methyl]carbamate (27e) (1.2 g, 2.67 mmol, 1.0 eq), 4,4-dihexoxybutanoic acid (27b) (1.93 g, 6.69 mmol, 2.5 eq), TEA (1.35 g, 13.37 mmol, 1.86 mL, 5.0 eq) and DMAP (980.31 mg, 8.02 mmol, 3.0 eq) in DCM (30 mL) was added EDCI (1.28 g, 6.69 mmol, 2.5 eq). The mixture was stirred at 25 °C for 12 h. The reaction mixture was diluted with H2O (20 mL), exacted with DCM 30 mL (15 mL × 2), and washed with brine 20 mL (10 mL × 2), dried over anhydrous sodium sulfate, filtered and concentrated under reduced pressure to give a residue. The residue was purified by flash silica gel chromatography (ISCO®; 40 g SepaFlash® Silica Flash Column, Eluent of 0 ~ 5% MeOH / DCM gradient @ 60 mL / min) to give [6- [benzyloxycarbonyl-[(1-methyl-4-piperidyl)methyl]amino]-11-(4,4- dihexoxybutanoyloxy)undecyl] 4,4-dihexoxybutanoate (27f) (1.88 g, 1.81 mmol, 67.5% yield, 95.0% purity) as a yellow oil. ¹H NMR (400 MHz, CDCl3) δ = 7.43 - 7.22 (m, 5H), 5.11 (s, 2H), 4.50 (t, J = 5.6 Hz, 2H), 4.15 - 3.95 (m, 4H), 3.61 - 3.52 (m, 4H), 3.47 - 3.37 (m, 4H), 3.03 - 2.94 (m, 2H), 2.88 - 2.76 (m, 2H), 2.38 (t, J = 7.6 Hz, 4H), 2.29 - 2.19 (m, 3H), 1.96 - 1.89 (m, 4H), 1.85 - 1.74 (m, 2H), 1.73 - 1.41 (m, 19H), 1.39 - 1.22 (m, 34H), 0.98 - 0.82 (m, 12H). [00477] Step 7: 6-(((1-methylpiperidin-4-yl)methyl)amino)undecane-1,11-diyl bis(4,4- bis(hexyloxy)butanoate) (27g)

27g [00478] To a solution of [6-[benzyloxycarbonyl-[(1-methyl-4-piperidyl)methyl]amino]-11- (4,4-dihexoxybutanoyloxy)undecyl] 4,4-dihexoxybutanoate (27f) (1.88 g, 1.90 mmol, 1.0 eq) in MeOH (15 mL) and THF (15 mL) were added Pd/C (10%, 100 mg) and Pd(OH)2/C (10%, 100 mg) under N₂. The suspension was degassed and purged with H₂3 times. The mixture was stirred under H2 (20 PSI ) at 40 °C for 3 h. The reaction mixture was filtered with DCM (50 mL) and MeOH (30 mL) and the filtrate concentrated under reduced pressure to give a residue. The residue was purified by flash silica gel chromatography (ISCO®; 20 g SepaFlash® Silica Flash Column, Eluent of 0 ~ 12% MeOH / DCM gradient @ 100 mL / min) to give [11-(4,4- dihexoxybutanoyloxy)-6-[(1-methyl-4-piperidyl)methylamino]undecyl] 4,4-dihexoxybutanoate (27g) (1.37 g, 1.52 mmol, 80.1% yield, 95.0% purity) as a yellow oil. ¹H NMR (400 MHz, CDCl3) δ = 4.53 - 4.44 (m, 2H), 4.05 (t, J = 6.8 Hz, 4H), 3.61 - 3.52 (m, 4H), 3.45 - 3.35 (m, 4H), 2.90 - 2.81 (m, 2H), 2.45 - 2.40 (m, 3H), 2.39 - 2.33 (m, 4H), 2.29 - 2.23 (m, 3H), 1.97 - 1.86 (m, 6H), 1.75 - 1.68 (m, 2H), 1.65 - 1.51 (m, 12H), 1.40 - 1.23 (m, 40H), 0.96 - 0.82 (m, 12H). [00479] Step 8: 6-((chlorocarbonyl)((1-methylpiperidin-4-yl)methyl)amino)undecane-1,11- diyl bis(4,4-bis(hexyloxy)butanoate) (27h)

[00480] To a solution of [11-(4,4-dihexoxybutanoyloxy)-6-[(1-methyl-4- piperidyl)methylamino]undecyl] 4,4-dihexoxybutanoate (27g) (500 mg, 584.58 μmol, 1.0 eq) and TEA (177.46 mg, 1.75 mmol, 244.10 μL, 3.0 eq) dissolved in dry DCM (10 mL) was added slowly bis(trichloromethyl) carbonate (104.08 mg, 350.75 μmol, 0.6 eq) at 0 °C under N₂. The resulting solution was stirred at 25 °C for 1 h (checked by TLC) and concentrated under reduced pressure and kept under N2. The reaction mixture was concentrated in vacuum to give [6- [chlorocarbonyl-[(1-methyl-4-piperidyl)methyl]amino]-11-(4,4-dihexoxybutanoyloxy)undecyl] 4,4-dihexoxybutanoate (27h) (536 mg, crude) as a yellow solid. [00481] Step 9: 6-(((heptylthio)carbonyl)((1-methylpiperidin-4-yl)methyl)amino)undecane- 1,11-diyl bis(4,4-bis(hexyloxy)butanoate) (27)

[00482] To a solution of heptane-1-thiol (231.74 mg, 1.75 mmol, 274.57 μL, 3.0 eq) dissolved in dry THF (10 mL) was added NaOH (70.08 mg, 1.75 mmol, 3.0 eq) at 0 °C under N₂. Then [6- [chlorocarbonyl-[(1-methyl-4-piperidyl)methyl]amino]-11-(4,4- dihexoxybutanoyloxy)undecyl]4,4-dihexoxybutanoate (27h) (536 mg, 584.02 μmol, 1.0 eq) dissolved in THF (5 mL) was added via syringe slowly under N₂ at 0 °C. The resulting solution was stirred at 25 °C for 4 h. The reaction mixture was quenched by NH₄Cl (10 mL) at 0 °C and then diluted with EtOAc (5 mL). The aqueous phase was extracted with EtOAc (10 mL × 2). The combined organic phase was washed with brine (10 mL), dried with anhydrous sodium sulfate, filtered and concentrated in vacuum to give residue. The residue was purified by prep- HPLC (FA condition; column: CD12-ACCHROM phenyl-Hexyl 150 × 25 × 10 μm; mobile phase: [water (FA) - ACN]; gradient: 54% - 84% B over 10 min) to give [11-(4,4- dihexoxybutanoyloxy)-6-[heptylsulfanylcarbonyl-[(1-methyl-4- piperidyl)methyl]amino]undecyl] 4,4-dihexoxybutanoate (27) (169.67 mg, 167.38 μmol, 28.66% yield, 99.99% purity) as a colorless oil. LCMS: [M+H]⁺: 1013.9¹H NMR (400 MHz, CDCl3) δ = 4.42 (t, J = 5.6 Hz, 2H), 3.98 (t, J = 6.8 Hz, 4H), 3.54 - 3.45 (m, 4H), 3.37 - 3.30 (m, 4H), 3.08 - 2.92 (m, 4H), 2.79 (t, J = 7.2 Hz, 2H), 2.38 - 2.24 (m, 7H), 1.88 - 1.83 (m, 4H), 1.80 - 1.62 (m, 4H), 1.58 - 1.45 (m, 16H), 1.42 - 1.34 (m, 4H), 1.32 - 1.16 (m, 42H), 0.85 - 0.77 (m, 15H). Example 28: Synthesis of [11-(4,4-dihexoxybutanoyloxy)-6-[(1-methyl-4-piperidyl)methyl- octylsulfinyl-amino]undecyl] 4,4-dihexoxybutanoate (28)

[00483] To a solution of [11-(4,4-dihexoxybutanoyloxy)-6-[(1-methyl-4- piperidyl)methylamino]undecyl] 4,4-dihexoxybutanoate (27g, 500 mg, 584.58 μmol, 1 eq) and TEA (177.46 mg, 1.75 mmol, 244.10 μL, 3 eq) in DCM (10 mL) was added dropwise octane-1- sulfinyl chloride (7b, 575.04 mg, 2.92 mmol, 5 eq) in DCM (5 mL) at 0 °C under N₂. The resulting mixture was stirred at 25 °C for 16 h. The reaction mixture was concentrated under reduced pressure to residue. The residue was purified by flash silica gel chromatography (20 g SepaFlash® Silica Flash Column, DCM : MeOH: 0~8%) to afford 28 (310 mg, 301.15 μmol, 51.8% yield, 98.66% purity) as yellow oil. LCMS: [M+H]⁺: 1015.9¹H NMR (400 MHz, CDCl3) δ = 4.50 (t, J = 5.6 Hz, 2H), 4.05 (dt, J = 2.4, 6.8 Hz, 4H), 3.62 - 3.54 (m, 4H), 3.46 - 3.37 (m, 5H), 3.17 - 3.05 (m, 1H), 3.01 - 2.94 (m, 1H), 2.92 - 2.85 (m, 1H), 2.84 - 2.72 (m, 2H), 2.67 (br s, 3H), 2.56 - 2.48 (m, 2H), 2.39 (dt, J = 3.2, 7.6 Hz, 4H), 1.98 - 1.90 (m, 6H), 1.88 - 1.80 (m, 2H), 1.68 - 1.59 (m, 8H), 1.59 - 1.53 (m, 8H), 1.44 - 1.25 (m, 46H), 0.93 - 0.87 (m, 15H). Example 29: Synthesis of 6-(((1-methylpiperidin-4-yl)methyl)(octylsulfinyl)amino)undecane- 1,11-diyl bis(6,6-bis(pentyloxy)hexanoate) (29) [00484] Step 1: 2-methoxycyclohexan-1-one oxime (29a)

[00485] To a solution of 2-methoxycyclohexanone (30 g, 234.07 mmol, 29.41 mL, 1.0 eq) in MeOH (200 mL) and H2O (300 mL) was added NaOAc (38.40 g, 468.13 mmol, 2.0 eq) followed by the addition of hydroxylamine hydrochloride (32.53 g, 468.13 mmol, 2.0 eq). The resulting mixture was heated at 70 °C and stirred for 16 h. The reaction mixture was concentrated under reduced pressure to remove MeOH. The residue was diluted with H2O (50 mL) and extracted with EtOAc (50 mL × 3). The combined organic layers washed with brine (60 mL), dried over anhydrous sodium sulfate, filtered and concentrated under reduced pressure to give 2-methoxycyclohexanone oxime (29a) (32.9 g, 229.78 mmol, 98.2% yield) as yellow oil. ¹H NMR (400 MHz, CDCl₃) δ = 3.72 (t, J = 3.6 Hz, 1H), 3.25 (s, 3H), 3.05 - 2.97 (m, 1H), 2.09 - 1.99 (m, 2H), 1.85 - 1.77 (m, 2H), 1.69 - 1.63 (m, 1H), 1.55 - 1.51 (m, 1H), 1.47 - 1.38 (m, 1H). [00486] Step 2: 6,6-dimethoxyhexanenitrile (29b)

[00487] To a solution of 2-methoxycyclohexanone oxime (29a) (10 g, 69.84 mmol, 1.0 eq) in DCM (120 mL) was added dropwise SOCl2 (9.97 g, 83.81 mmol, 6.09 mL, 1.2 eq) at 0 °C under N2. After addition, the mixture was stirred at 0 °C for 0.5 h, and then MeOH (29.70 g, 926.79 mmol, 37.50 mL, 13.3 eq) was added dropwise at 0 °C. The resulting mixture was stirred at 20 °C for 15.5 h. The reaction mixture was concentrated in vacuum to give residue, The residue was purified by flash silica gel chromatography (20 g SepaFlash® Silica Flash Column, PE : EtOAc = 0 ~ 30%) to give 6,6-dimethoxyhexanenitrile (29b) (2.91 g, 18.51 mmol, 26.5% yield) as colorless oil. ¹H NMR (400 MHz, CDCl3) δ = 4.33 (t, J = 5.6 Hz, 1H), 3.29 (s, 6H), 2.30 (t, J = 7.6 Hz, 2H), 1.65 - 1.57 (m, 4H), 1.42 - 1.30 (m, 2H). [00488] Step 3: 6,6-bis(pentyloxy)hexanenitrile (29c)

[00489] A mixture of 6,6-dimethoxyhexanenitrile (29b) (4.0 g, 25.44 mmol, 1.0 eq), PPTS (3.20 g, 12.72 mmol, 0.5 eq) and pentan-1-ol (11.21 g, 127.22 mmol, 13.83 mL, 5.0 eq) was degassed and purged with N₂3 times, and then the mixture was stirred at 110 °C for 16 h under N2. The reaction was concentrated to afforded residue. The residue was purified by flash silica gel chromatography (ISCO®; 120 g SepaFlash® Silica Flash Column, Eluent of 0 ~ 2% EtOAc / PE gradient @ 50 mL / min) to give 6,6-dipentoxyhexanenitrile (29c) (2.92 g, 9.75 mmol, 38.3% yield, 90% purity) as a yellow oil. ¹H NMR (400 MHz, CDCl3) δ = 4.45 (t, J = 5.6 Hz, 1H), 3.60 - 3.50 (m, 2H), 3.43 - 3.35 (m, 2H), 2.34 - 2.26 (m, 2H), 1.67 - 1.59 (m, 6H), 1.39 - 1.30 (m, 12H), 0.92 - 0.88 (m, 6H). [00490] Step 4: 6,6-bis(pentyloxy)hexanoic acid (29d)

[00491] A mixture of 6,6-dipentoxyhexanenitrile (29c) (3.70 g, 13.73 mmol, 1.0 eq) and KOH (2.31 g, 41.20 mmol, 3.0 eq) in EtOH (15 mL) and H₂O (15 mL) was degassed and purged with N23 times, and then the mixture was stirred at 110 °C for 10 h under N2. The reaction mixture cooled down to room temperature and concentrated, pH adjust to pH = 4 by addition 1N HCl, then the mixture was extracted with EtOAc 60 mL (30 mL × 2). The combined organic layers washed with brine (50 mL), dried over anhydrous sodium sulfate, filtered and concentrated under reduced pressure to give a residue. The residue was purified by flash silica gel chromatography (ISCO®; 120 g SepaFlash® Silica Flash Column, Eluent of 0 ~ 10% EtOAc / PE gradient @ 100 mL / min) to give 6,6-dipentoxyhexanoic acid (29d) (2.31 g, 7.21 mmol, 52.5% yield, 90% purity) as a yellow oil.¹H NMR (400 MHz, CDCl3) δ = 4.44 (t, J = 5.6 Hz, 1H), 3.57 - 3.49 (m, 2H), 3.42 - 3.33 (m, 2H), 2.33 (t, J = 7.6 Hz, 2H), 1.65 - 1.51 (m, 8H), 1.44 - 1.37 (m, 2H), 1.33 - 1.27 (m, 8H), 0.91 - 0.84 (m, 6H). [00492] Step 5: 6-(((benzyloxy)carbonyl)((1-methylpiperidin-4-yl)methyl)amino)undecane-

[00493] To a solution of benzyl N-[6-hydroxy-1-(5-hydroxypentyl)hexyl]-N-[(1-methyl-4- piperidyl)methyl]carbamate (27e) (1.4 g, 3.12 mmol, 1.0 eq), 6,6-dipentoxyhexanoic acid (29d) (2.25 g, 7.80 mmol, 2.5 eq), TEA (1.58 g, 15.60 mmol, 2.17 mL, 5.0 eq) and DMAP (1.14 g, 9.36 mmol, 3.0 eq) in DCM (35 mL) was added EDCI (1.50 g, 7.80 mmol, 2.5 eq), and the mixture was stirred at 25 °C for 12 h. The reaction mixture was diluted with H2O (20 mL). exacted with DCM 30 mL (10 mL × 3) washed with brine 20 mL (10 mL × 2), dried over anhydrous sodium sulfate, filtered and concentrated under reduced pressure to give a residue. The residue was purified by flash silica gel chromatography (ISCO®; 40 g SepaFlash® Silica Flash Column, Eluent of 0 ~ 5% MeOH / DCM gradient @ 60 mL / min) to give [6- [benzyloxycarbonyl-[(1-methyl-4-piperidyl)methyl]amino]-11-(6,6- dipentoxyhexanoyloxy)undecyl] 6,6-dipentoxyhexanoate (29e) (1.72 g, 1.65 mmol, 52.9% yield, 95% purity) as a yellow oil. ¹H NMR (400 MHz, CDCl3) δ = 7.48 - 7.14 (m, 4H), 5.11 (s, 2H), 4.46 (t, J = 5.6 Hz, 2H), 4.11 - 3.94 (m, 4H), 3.59 - 3.51 (m, 4H), 3.46 - 3.36 (m, 4H), 3.06 - 2.95 (m, 2H), 2.92 - 2.76 (m, 2H), 2.34 - 2.17 (m, 7H), 1.94 - 1.75 (m, 4H), 1.71 - 1.49 (m, 28H), 1.40 - 1.24 (m, 28H), 0.95 - 0.83 (m, 12H). [00494] Step 6: 6-(((1-methylpiperidin-4-yl)methyl)amino)undecane-1,11-diyl bis(6,6- bis(pentyloxy)hexanoate) (29f)

[00495] To a solution of [6-[benzyloxycarbonyl-[(1-methyl-4-piperidyl)methyl]amino]-11- (6,6-dipentoxyhexanoyloxy)undecyl] 6,6-dipentoxyhexanoate (29e) (1.72 g, 1.74 mmol, 1.0 eq) in THF (15 mL) and MeOH (15 mL) were added Pd/C (10%, 150 mg) and Pd(OH)₂/C (10%, 150 mg) under N2. The suspension was degassed and purged with H23 times. The mixture was stirred under H₂ (20 PSI) at 40 °C for 2 h. The reaction mixture was filtered with DCM (50 mL) and MeOH (30 mL) and the filtrate concentrated under reduced pressure to give a residue. The residue was purified by flash silica gel chromatography (ISCO®; 20 g SepaFlash® Silica Flash Column, Eluent of 0 ~ 15% EtOAc / PE gradient @ 100 mL / min) to give [11-(6,6- dipentoxyhexanoyloxy)-6-[(1-methyl-4-piperidyl)methylamino]undecyl] 6,6- dipentoxyhexanoate (29f) (1.18 g, 1.31 mmol, 75.4% yield, 95% purity) as a yellow oil. ¹H NMR (400 MHz, CDCl₃) δ = 4.46 (t, J = 5.6 Hz, 2H), 4.06 (t, J = 6.8 Hz, 4H), 3.62 - 3.52 (m, 4H), 3.45 - 3.35 (m, 4H), 2.95 - 2.82 (m, 2H), 2.47 - 2.39 (m, 3H), 2.35 - 2.26 (m, 7H), 2.01 - 1.88 (m, 2H), 1.78 - 1.71 (m, 2H), 1.68 - 1.54 (m, 20H), 1.44 - 1.25 (m, 36H), 0.98 - 0.85 (m, 12H). [00496] Step 7: 6-(((1-methylpiperidin-4-yl)methyl)(octylsulfinyl)amino)undecane-1,11-diyl bis(6,6-bis(pentyloxy)hexanoate) (29)

[00497] To a solution of [11-(6,6-dipentoxyhexanoyloxy)-6-[(1-methyl-4- piperidyl)methylamino]undecyl] 6,6-dipentoxyhexanoate (29f) (300 mg, 350.75 μmol, 1.0 eq) and TEA (177.46 mg, 1.75 mmol, 244.10 μL, 5.0 eq) in DCM (5 mL) was added octane-1- sulfinyl chloride (7b) (345.02 mg, 1.75 mmol, 5.0 eq) in DCM (0.3 mL) at 0 °C. The mixture was stirred at 25 °C for 5 h. The reaction was concentrated and purified by flash silica gel chromatography (ISCO®; 12 g SepaFlash® Silica Flash Column, Eluent of 0 ~ 5% MeOH / DCM gradient @ 20 mL / min) to give [11-(6,6-dipentoxyhexanoyloxy)-6-[(1-methyl-4- piperidyl)methyl-octylsulfinyl-amino]undecyl] 6,6-dipentoxyhexanoate (29) (106.41 mg, 103.40 μmol, 29.5% yield, 98.69% purity) as a yellow oil. LCMS: [M+H]⁺: 1016.2¹H NMR (400 MHz, CDCl3) δ = 4.46 (t, J = 5.6 Hz, 2H), 4.09 - 4.01 (m, 4H), 3.60 - 3.51 (m, 4H), 3.45 - 3.36 (m, 4H), 3.11 - 3.01 (m, 2H), 2.99 - 2.92 (m, 1H), 2.77 - 2.65 (m, 2H), 2.58 - 2.49 (m, 1H), 2.39 (s, 3H), 2.34 - 2.27 (m, 4H), 2.17 - 2.01 (m, 2H), 1.86 - 1.71 (m, 4H), 1.65 - 1.50 (m, 24H), 1.45 - 1.22 (m, 42H), 0.97 - 0.83 (m, 15H). Example 30: Synthesis of bis(2-butyloctyl) 10-[heptoxycarbonyl-[(1-methyl-4- piperidyl)methyl]amino]nonadecanedioate (30) [00498] Step 1: heptyl (4-nitrophenyl) carbonate (30a)

[00499] To a solution of heptan-1-ol (2 g, 17.21 mmol, 2.43 mL, 1 eq) and pyridine (1.36 g, 17.21 mmol, 1.39 mL, 1 eq) in DCM (30 mL) were added (4-nitrophenyl) carbonochloridate (5.20 g, 25.82 mmol, 1.5 eq) and DMAP (2.10 g, 17.21 mmol, 1 eq) at 0 °C under N2. After addition, the mixture was stirred at 25 °C for 16 h. The reaction mixture was concentrated under reduced pressure to give residue. The residue was purified by flash silica gel chromatography (40 g SepaFlash® Silica Flash Column, PE : EtOAc: 0~10%) to give heptyl (4-nitrophenyl) carbonate (30a, 2.8 g, 9.95 mmol, 57.8% yield) as colorless oil. ¹H NMR (400 MHz, CDCl₃) δ = 8.31 - 8.26 (m, 2H), 7.42 - 7.36 (m, 2H), 4.30 (t, J = 6.8 Hz, 2H), 1.83 - 1.73 (m, 2H), 1.46 - 1.30 (m, 8H), 0.91 (t, J = 6.4 Hz, 3H). [00500] Step 2: bis(2-butyloctyl) 10-[heptoxycarbonyl-[(1-methyl-4- piperidyl)methyl]amino]nonadecanedioate (30)

[00501] To a solution of bis(2-butyloctyl) 10-[(1-methyl-4- piperidyl)methylamino]nonadecanedioate (5f, 500 mg, 631.86 μmol, 1 eq) and DMAP (77.19 mg, 631.86 μmol, 1 eq) in DCM (10 mL) were added pyridine (99.96 mg, 1.26 mmol, 102.00 μL, 2 eq) and heptyl (4-nitrophenyl) carbonate (30a, 444.36 mg, 1.58 mmol, 2.5 eq). The mixture was stirred at 45 °C for 48 h. The reaction mixture was concentrated under reduced pressure to give residue. The residue was purified by prep-HPLC (column: PHS-Phenyl-Hexyl 150 × 25mm × 7 μm; mobile phase: [water (FA) - ACN]; gradient: 58% - 88% B over 10 min) to give (30) (378 mg, 397.96 μmol, 64.1% yield, 98.28% purity) as yellow oil. LCMS: [M+H] ⁺: 933.9¹H NMR (400 MHz, CD₃OD-d₄) δ = 4.10 (t, J = 6.4 Hz, 4H), 4.06 (t, J = 6.4 Hz, 2H), 4.00 (d, J = 5.6 Hz, 4H), 3.07 - 3.03 (m, 2H), 2.96 - 2.90 (m, 2H), 2.34 - 2.28 (m, 7H), 2.08 - 1.98 (m, 2H), 1.68 - 1.60 (m, 12H), 1.36 - 1.28 (m, 62H), 0.92 - 0.89 (m, 15H). Example 31: Synthesis of [11-(6,6-dipentoxyhexanoyloxy)-6-[(1-methyl-4-piperidyl)methyl- nonanoyl-amino]undecyl] 6,6-dipentoxyhexanoate (31)

[00502] To a solution of [11-(6,6-dipentoxyhexanoyloxy)-6-[(1-methyl-4- piperidyl)methylamino]undecyl] 6,6-dipentoxyhexanoate (29f, 300 mg, 350.75 μmol, 1 eq) and TEA (70.98 mg, 701.49 μmol, 97.64 μL, 2 eq) in DCM (5 mL) was added dropwise nonanoyl chloride (92.96 mg, 526.12 μmol, 98.89 μL, 1.5 eq) in DCM (2 mL) at 0 °C under N2. The resulting mixture was stirred at 20 °C for 2 hr. The reaction mixture was concentrated under reduced pressure to residue. The residue was purified by prep-HPLC (column: CD12- ACCHROM phenyl-Hexyl 150 × 25 × 10 um; mobile phase: [water (formic acid) - ACN]; gradient: 54% - 84% B over 10 min) and then purified by flash silica gel chromatography (12 g SepaFlash® Silica Flash Column, DCM : MeOH: 0~7%) to afford [11-(6,6- dipentoxyhexanoyloxy)-6-[(1-methyl-4-piperidyl)methyl-nonanoyl-amino]undecyl] 6,6- dipentoxyhexanoate (31) (198 mg, 198.87 μmol, 56.7% yield, 99.99% purity) as yellow oil. LCMS: [M+H]⁺: 996.1¹H NMR (400 MHz, CDCl3) δ = 4.38 (t, J = 5.6 Hz, 2H), 4.03 - 3.93 (m, 4H), 3.61 - 3.53 (m, 1H), 3.53 - 3.45 (m, 4H), 3.37 - 3.29 (m, 4H), 3.24 - 3.13 (m, 1H), 3.04 - 2.88 (m, 2H), 2.51 (br s, 3H), 2.37 - 2.30 (m, 1H), 2.27 - 2.20 (m, 6H), 1.62 - 1.45 (m, 27H), 1.41 - 1.13 (m, 44H), 0.89 - 0.75 (m, 15H). Example 32: Synthesis of bis(2-butyloctyl) 10-(2,2-difluoro-N-((1-methylpiperidin-4- yl)methyl)nonanamido)nonadecanedioate (32) [00503] Step 1: 2,2-difluorononanal (32a)

[00504] To a round-bottom flask equipped with a magnetic stir bar and charged with pyrrolidine-2-carboxylic acid (971.30 mg, 8.44 mmol, 0.3 eq) and NFSI (44.34 g, 140.61 mmol, 5.0 eq) in THF (120 mL). The mixture was stirred at 20 °C until all solids were dissolved and the nonanal (4 g, 28.12 mmol, 1.0 eq) was then slowly added to the reaction mixture dissolved in THF (20 mL) and the mixture was stirred for 16 h. The reaction diluted with MTBE (100 mL) and cooled down to -78 °C, filtered through a pad of silica gel, eluting with cold MTBE (100 mL × 6), The resultant organic layer was washed with saturated NaHCO₃ (300), brine (100 mL), dried over anhydrous sodium sulfate, filtered and concentrated. The residue was purified by flash silica gel chromatography (ISCO®; 120 g SepaFlash® Silica Flash Column, Eluent of 0 ~ 15% EtOAc / PE gradient @ 50 mL / min) to give compound 2,2-difluorononane-1,1-diol (32a) (3.8 g, 19.36 mmol, 68.86% yield) as a white solid. ¹H NMR (400 MHz, DMSO-d6) δ = 7.18 - 7.03 (m, 1H), 6.44 (d, J = 6.4 Hz, 1H), 4.97 - 4.67 (m, 1H), 1.94 - 1.72 (m, 2H), 1.48 - 1.38 (m, 2H), 1.28 - 1.22 (m, 8H), 0.87 - 0.84 (m, 3H). [00505] Step 2: 2,2-difluorononanoic acid (32b):

[00506] To a solution of 2,2-difluorononane-1,1-diol (32a) (1 g, 5.61 mmol, 1.0 eq) in t-BuOH (30 mL) was added 2-methylbut-2-ene (2.75 g, 39.28 mmol, 4.16 mL, 7.0 eq), a solution of NaClO₂ (1.01 g, 11.22 mmol, 2.0 eq) and NaH₂PO₄ (2.02 g, 16.83 mmol, 3.0 eq) in H₂O (15 mL). The mixture was stirred at 25 °C for 2 h. the mixture was quenched with 1 N HCl adjust pH to 3 - 4 and extracted with EtOAc (15 mL × 3). The organic phase was washed with brine (20 mL × 3) and dried over anhydrous sodium sulfate. filtered and concentrated. The residue was purified by flash silica gel chromatography (ISCO®; 20 g SepaFlash® Silica Flash Column, Eluent of 0 ~ 8% MeOH / DCM gradient @100 mL / min) to give compound 2,2- difluorononanoic acid (32b) (866 mg, 4.46 mmol, 79.5% yield) as a colorless oil.¹H NMR (400 MHz, DMSO-d6) δ = 14.60 - 11.58 (m, 1H), 2.11 - 1.98 (m, 2H), 1.44 - 1.22 (m, 10H), 0.85 (t, J = 6.8 Hz, 3H). [00507] Step 3: 2,2-difluorononanoyl chloride (32c): [00508] A mixture of 2,2-difluorononanoic acid (32b) (620 mg, 3.19 mmol, 1.0 eq) in DCM (6 mL) was degassed and purged with N23 times, then SOCl2 (1.14 g, 9.58 mmol, 695.58 μL, 3.0 eq) was added slowly, and one drop DMF was added followed at 25 °C. the mixture was stirred at 40 °C for 2 h under N₂. The reaction was concentrated to give compound 2,2- difluorononanoyl chloride (32c) (655 mg, crude) as a colorless oil. [00509] Step 4: bis(2-butyloctyl) 10-(2,2-difluoro-N-((1-methylpiperidin-4- yl)methyl)nonanamido)nonadecanedioate (32)

[00510] To a solution of bis(2-butyloctyl) 10-[(1-methyl-4- piperidyl)methylamino]nonadecanedioate (5f) (600 mg, 758.23 μmol, 1.0 eq), 2,2- difluorononanoyl chloride (32c) (644.99 mg, 3.03 mmol, 4.0 eq) in DCM (8 mL) was added TEA (383.62 mg, 3.79 mmol, 527.68 μL, 5.0 eq). The mixture was stirred at 25 °C for 15 min. The reaction mixture was partitioned between DCM (10 mL) and H2O (20 mL). The organic phase was separated, washed with brine 20 mL (10 mL × 2), dried over anhydrous sodium sulfate, filtered and concentrated under reduced pressure to give a residue. The residue was purified by flash silica gel chromatography (ISCO®; 40 g SepaFlash® Silica Flash Column, Eluent of 0 ~ 5% MeOH / DCM gradient @ 60 mL / min).then was further purified by prep- HPLC (FA condition; column: CD27-PHS XP phenyl-Hexyl 150 × 25 × 7 um; mobile phase: [water (FA) - ACN]; gradient: 60% - 90% B over 10 min) to give compound bis(2-butyloctyl) 10-[2,2-difluorononanoyl-[(1-methyl-4-piperidyl)methyl]amino]nonadecanedioate (32) (118.94 mg, 122.92 μmol, 39.6% yield, 99.99% purity) as a yellow gum. LCMS: [M+H]⁺: 968.0¹H NMR (400 MHz, CDCl3) δ = 4.11 - 4.01 (m, 1H), 3.97 (d, J = 6.0 Hz, 4H), 3.24 - 3.12 (m, 2H), 3.11 - 3.02 (m, 2H), 2.51 (s, 3H), 2.44 - 2.35 (m, 1H), 2.34 - 2.27 (m, 5H), 2.18 - 2.06 (m, 2H), 2.01 - 1.90 (m, 1H), 1.83 - 1.73 (m, 2H), 1.67 - 1.55 (m, 8H), 1.51 - 1.43 (m, 4H), 1.38 - 1.16 (m, 62H), 0.97 - 0.82 (m, 15H). Example 33: 8-(((1-methylpiperidin-4-yl)methyl)(octylsulfinyl)amino)pentadecane-1,15-diyl bis(4-butyldecanoate) (33) [00511] Step 1: 2-butyloctanal (33a)

[00512] A mixture of 2-butyloctan-1-ol (15 g, 80.50 mmol, 1.0 eq), PCC (34.70 g, 161.00 mmol, 2.0 eq) and silica gel (34.71 g, 577.69 mmol, 7.18 eq) in DCM (300 mL) was degassed and purged with N23 times, and then the mixture was stirred at 25 °C for 2 h under N2. The mixture was filtered through a short silica gel column and washed with hexane / MBTE = 20 / 1 to give compound 2-butyloctanal (33a) (15.43 g, crude) as a green oil. ¹H NMR (400 MHz, DMSO-d6) δ = 9.61 - 9.38 (m, 1H), 2.30 - 2.10 (m, 1H), 1.57 - 1.32 (m, 4H), 1.29 - 1.14 (m, 12H), 0.89 - 0.81 (m, 6H). [00513] Step 2: methyl (E)-4-butyldec-2-enoate (33b)

[00514] A mixture of methyl 2-(triphenyl-phosphanylidene)acetate (19.95 g, 59.68 mmol, 1.0 eq) and 2-butyloctanal (33a) (11 g, 59.68 mmol, 1.0 eq) in THF (120 mL) was degassed and purged with N₂3 times, and then the mixture was stirred at 70 °C for 12 h under N₂. the reaction mixture concentrated under reduced pressure and then taken up in DCM (20 mL). MTBE was slowly added until precipitation appeared. The white suspension was then filtered through a pad of Celite. The filtrate was concentrated under reduced pressure. The crude product was purified by flash chromatography on silica gel (PE : EtOAc = 1 / 0 gradient to 100 / 1) to give compound methyl (E)-4-butyldec-2-enoate (33b) (7.94 g, 24.77 mmol, 41.5% yield, 75% purity) as a colorless oil. ¹H NMR (400 MHz, CDCl₃) δ = 6.82 - 6.69 (m, 1H), 5.82 - 5.71 (m, 1H), 3.73 (s, 3H), 2.29 - 2.17 (m, 1H), 1.47 - 1.40 (m, 4H), 1.29 - 1.27 (m, 8H), 1.26 - 1.21 (m, 10H), 0.91 - 0.88 (m, 6H). [00515] Step 3: methyl 4-butyldecanoate (33c):

[00516] To a solution of methyl (E)-4-butyldec-2-enoate (33b) (7.94 g, 33.03 mmol, 1.0 eq) in THF (40 mL) and MeOH (40 mL) was added Pd/C (10%, 1.0 g) under N₂. The suspension was degassed and purged with H₂3 times. The mixture was stirred under H₂ (15 Psi) at 20 °C for 10 h. The reaction mixture was filtered with DCM (50 mL) and MeOH (30 mL) and the filtrate concentrated under reduced pressure to give compound methyl 4-butyldecanoate (33c) (7.18 g, 14.81 mmol, 44.8% yield, 50% purity) as a yellow oil. [00517] Step 4: 4-butyldecanoic acid (33d):

[00518] A mixture of methyl 4-butyldecanoate (33c) (7.18 g, 29.62 mmol, 1.0 eq) and LiOH•H₂O (2.49 g, 59.24 mmol, 2.0 eq) in THF (80 mL) and H₂O (16 mL) was degassed and purged with N23 times, and then the mixture was stirred at 25 °C for 10 h under N2. The reaction mixture was cooled down to room temperature and concentrated, the mixture was diluted with EtOAc (60 mL), pH adjusted to pH = 4 by addition 1N HCl, and extracted with EtOAc 60 mL (30 mL × 2). The combined organic layers were washed with brine (50 mL), dried over anhydrous sodium sulfate, filtered and concentrated under reduced pressure to give a residue. The residue was purified by flash silica gel chromatography (ISCO®; 120 g SepaFlash® Silica Flash Column, Eluent of 0 ~ 10% EtOAc / PE gradient @ 100 mL / min) to give compound 4-butyldecanoic acid (33d) (3.75 g, 14.78 mmol, 49.9% yield, 90% purity) as a yellow oil. ¹H NMR (400 MHz, CDCl₃) δ = 2.41 - 2.27 (m, 2H), 1.71 - 1.54 (m, 2H), 1.40 - 1.18 (m, 17H), 0.95 - 0.84 (m, 6H). [00519] Step 5: dimethyl 8-(((1-methylpiperidin-4-yl)methyl)amino)pentadecanedioate (33e)

, 5 , 0

33e [00520] A mixture of (1-methyl-4-piperidyl)methanamine (2.5 g, 19.50 mmol, 1.0 eq), NaBH(OAc)3 (6.20 g, 29.25 mmol, 1.5 eq), HOAc (1.76 g, 29.25 mmol, 1.67 mL, 1.5 eq) in DCM (40 mL) was degassed and purged with N₂3 times, dimethyl 8-oxopentadecanedioate (6.74 g, 21.45 mmol, 1.1 eq) in DCM (30 mL) was added and then the mixture was stirred at 25 °C for 10 h under N2. The mixture pH was adjusted to 9 - 10 was with 1 N NaOH (20 mL), the aqueous phase was extracted with EtOAc (35 mL × 3). The combined organic phase was washed with brine (40 mL), dried with anhydrous sodium sulfate, filtered and concentrated in vacuum to give crude product. The residue was purified by flash silica gel chromatography (ISCO®; 80 g SepaFlash® Silica Flash Column, Eluent of 0 ~ 5% MeOH / DCM @ 100 mL / min) to give compound dimethyl 8-[(1-methyl-4-piperidyl)methylamino]pentadecanedioate (33e) (5.22 g, 11.62 mmol, 59.6% yield, 95% purity) was obtained as a yellow oil. ¹H NMR (400 MHz, CDCl3) δ = 3.64 (s, 6H), 2.82 (d, J = 11.6 Hz, 2H), 2.43 - 2.34 (m, 3H), 2.28 (t, J = 7.6 Hz, 4H), 2.23 (s, 3H), 1.93 - 1.83 (m, 2H), 1.72 - 1.65 (m, 2H), 1.64-1.54 (m, 4H), 1.36 - 1.18 (m, 20H). [00521] Step 6: dimethyl 8-(((benzyloxy)carbonyl)((1-methylpiperidin-4- yl)methyl)amino)pentadecanedioate (33f)

[00522] A mixture of dimethyl 8-[(1-methyl-4-piperidyl)methylamino]pentadecanedioate (33e) (3 g, 7.03 mmol, 1.0 eq), DIEA (1.82 g, 14.06 mmol, 2.45 mL, 2.0 eq) in DCM (50 mL) was degassed and purged with N23 times, then benzyl (2,5-dioxopyrrolidin-1-yl) carbonate (2.10 g, 8.44 mmol, 1.2 eq) was added at 0 °C, and the reaction was stirred at 25 °C for 3 h. The reaction mixture was concentrated under reduced pressure to give a residue. The residue was purified by flash silica gel chromatography (ISCO®; 80 g SepaFlash® Silica Flash Column, Eluent of 0 ~ 5% MeOH / DCM gradient @ 100 mL / min) to give compound dimethyl 8- [benzyloxycarbonyl-[(1-methyl-4-piperidyl)methyl]amino]pentadecanedioate (33f) (4.08 g, 6.91 mmol, 98.3% yield, 95% purity) as a yellow oil. ¹H NMR (400 MHz, CDCl3) δ = 7.39 - 7.27 (m, 5H), 5.09 (s, 2H), 3.81 - 3.57 (m, 7H), 3.41 - 3.19 (m, 2H), 3.08 - 2.94 (m, 2H), 2.66 - 2.56 (m, 7H), 2.54 - 2.41 (m, 2H), 2.33 - 2.20 (m, 5H), 1.89 - 1.70 (m, 2H), 1.62-1.52 (m, 6H), 1.29 - 1.14 (m, 12H). [00523] Step 7: benzyl (1,15-dihydroxypentadecan-8-yl)((1-methylpiperidin-4- yl)methyl)carbamate (33g)

[00524] A mixture of dimethyl 8-[benzyloxycarbonyl-[(1-methyl-4- piperidyl)methyl]amino]pentadecanedioate (33f) (3.88 g, 6.92 mmol, 1.0 eq) in THF (60 mL) was degassed and purged with N₂3 times and cooled down to 0 °C, then LiAlH₄ (2.5 M, 5.81 mL, 2.1 eq) was added via injection at 0 °C, and then the mixture was stirred at 25 °C for 1 h under N2. After completion, the reaction mixture was diluted with THF (2 mL), then successively was added H₂O (0.55 mL), aq. NaOH (0.55 mL, 15%), H₂O (1.65 mL) and dried over Na2SO4 at 0 °C under N2. The reaction mixture was filtered and the filtrate was concentrated in vacuum to give residue. The residue was purified by flash silica gel chromatography (ISCO®; 40 g SepaFlash® Silica Flash Column, Eluent of 0 ~ 5% MeOH / DCM gradient @ 50 mL / min) to give compound benzyl N-[8-hydroxy-1-(7- hydroxyheptyl)octyl]-N-[(1-methyl-4-piperidyl)methyl]carbamate (33g) (2.32 g, 4.14 mmol, 59.9% yield, 90% purity) as a colorless oil. ¹H NMR (400 MHz, CDCl₃) δ = 7.42 - 7.25 (m, 5H), 5.11 (s, 2H), 3.67 - 3.58 (m, 4H), 3.07 - 2.93 (m, 2H), 2.90 - 2.76 (m, 2H), 2.28 - 2.19 (m, 3H), 1.97 - 1.73 (m, 6H), 1.59 - 1.44 (m, 8H), 1.36 - 1.15 (m, 18H). [00525] Step 8: 8-(((benzyloxy)carbonyl)((1-methylpiperidin-4-yl)methyl)amino)pentadecane- 1,15-diyl bis(4-butyldecanoate) (33h)

[00526] To a solution of benzyl N-[8-hydroxy-1-(7-hydroxyheptyl)octyl]-N-[(1-methyl-4- piperidyl)methyl]carbamate (33g) (2.29 g, 4.54 mmol, 1.0 eq), 4-butyldecanoic acid (33d) (2.59 g, 11.34 mmol, 2.5 eq), TEA (2.30 g, 22.68 mmol, 3.16 mL, 5.0 eq) and DMAP (1.66 g, 13.61 mmol, 3.0 eq) in DCM (50 mL) was added EDCI (2.17 g, 11.34 mmol, 2.5 eq). The mixture was stirred at 25 °C for 16 h. The reaction mixture was diluted with H2O (60 mL) and extracted with DCM 80 mL (40 mL × 2). The combined organic layers were washed with brine 60 mL (30 mL × 2), dried over anhydrous sodium sulfate, filtered and concentrated under reduced pressure to give a residue. The residue was purified by flash silica gel chromatography (ISCO®; 80 g SepaFlash® Silica Flash Column, Eluent of 0 ~ 5% MeOH / DCM gradient @ 80 mL / min) to give compound [8-[benzyloxycarbonyl-[(1-methyl-4-piperidyl)methyl]amino]-15-(4- butyldecanoyloxy)pentadecyl] 4-butyldecanoate (33h) (1.74 g, 1.69 mmol, 37.3% yield, 90% purity) as a yellow oil.¹H NMR (400 MHz, CDCl3) δ = 7.39 - 7.27 (m, 5H), 5.11 (s, 2H), 4.04 (t, J = 6.8 Hz, 4H), 3.86 - 3.64 (m, 1H), 2.90 - 2.74 (m, 2H), 2.31 - 2.20 (m, 7H), 1.93 - 1.84 (m, 3H), 1.84 - 1.75 (m, 1H), 1.72 - 1.63 (m, 2H), 1.62 - 1.56 (m, 8H), 1.54 - 1.40 (m, 4H), 1.34 - 1.17 (m, 52H), 0.98 - 0.80 (m, 12H). [00527] Step 9: 8-(((1-methylpiperidin-4-yl)methyl)amino)pentadecane-1,15-diyl bis(4- butyldecanoate) (33i)

[00528] To a solution of [8-[benzyloxycarbonyl-[(1-methyl-4-piperidyl)methyl]amino]-15-(4- butyldecanoyloxy)pentadecyl] 4-butyldecanoate (33h) (1.74 g, 1.88 mmol, 1.0 eq) in MeOH (20 mL) and THF (20 mL) were added Pd/C (10%, 300 mg) and Pd(OH)2/C (20%, 300 mg) under N2. The suspension was degassed and purged with H23 times. The mixture was stirred under H2 (20 Psi) at 40 °C for 2 h. The reaction mixture was filtered with DCM (50 mL) and MeOH (30 mL) and the filtrate concentrated under reduced pressure to give a residue. The residue was purified by flash silica gel chromatography (ISCO®; 40 g SepaFlash® Silica Flash Column, Eluent of 0 ~ 5% MeOH / DCM gradient @ 100 mL / min) to give compound [15-(4- butyldecanoyloxy)-8-[(1-methyl-4-piperidyl)methylamino]pentadecyl] 4-butyldecanoate (33i) (1.53 g, 1.84 mmol, 97.7% yield, 95% purity) as a yellow oil. ¹H NMR (400 MHz, CDCl3) δ = 4.05 (t, J = 6.8 Hz, 4H), 2.90 - 2.78 (m, 2H), 2.47 - 2.35 (m, 3H), 2.31 - 2.23 (m, 7H), 1.96 - 1.85 (m, 2H), 1.76 - 1.69 (m, 2H), 1.63 - 1.55 (m, 8H), 1.40 - 1.14 (m, 58H), 0.95 - 0.79 (m, 12H). [00529] Step 10: 8-(((1-methylpiperidin-4-yl)methyl)(octylsulfinyl)amino)pentadecane-1,15- diyl bis(4-butyldecanoate) (33)

[00530] To a solution of [15-(4-butyldecanoyloxy)-8-[(1-methyl-4- piperidyl)methylamino]pentadecyl] 4-butyldecanoate (33i) (600 mg, 758.23 μmol, 1.0 eq), DMAP (27.79 mg, 227.47 μmol, 0.3 eq) and TEA (230.17 mg, 2.27 mmol, 316.61 μL, 3.0 eq) in DCM (10 mL) was added octane-1-sulfinyl chloride (745.86 mg, 3.79 mmol, 5.0 eq) in DCM (2 mL) at 0 °C. The mixture was stirred at 25 °C for 2 h. The reaction was concentrated. The residue was purified by flash silica gel chromatography (ISCO®; 20 g SepaFlash® Silica Flash Column, Eluent of 0 ~ 4% MeOH / DCM gradient @ 35 mL / min) to give compound [15-(4- butyldecanoyloxy)-8-[(1-methyl-4-piperidyl)methyl-octylsulfinyl-amino]pentadecyl] 4- butyldecanoate (33) (323.6 mg, 340.03 μmol, 44.8% yield, 99.99% purity) as a yellow gum. LCMS: [M+H]⁺: 951.9¹H NMR (400 MHz, CDCl3) δ = 4.10 - 3.97 (m, 4H), 3.52 - 3.33 (m, 2H), 3.13 - 3.03 (m, 1H), 3.02 -2.92(m, 1H), 2.90 - 2.80 (m, 1H), 2.79 - 2.73 (m, 1H), 2.72 - 2.67 (m, 3H), 2.66 - 2.55 (m, 2H), 2.54 - 2.47 (m, 1H), 2.30 - 2.21 (m, 4H), 2.03 - 1.87 (m, 2H), 1.86 - 1.67 (m, 3H), 1.64 - 1.53 (m, 10H), 1.50 - 1.40 (m, 4H), 1.36 - 1.13 (m, 60H), 0.97 - 0.80 (m, 15H). Example 34: bis(2-butyloctyl) 10-[(2-hexoxy-2-oxo-ethyl)-[(1-methyl-4- piperidyl)methyl]amino]nonadecanedioate (34) [00531] Step 1: hexyl 2-bromoacetate (34a)

[00532] To a solution of hexan-1-ol (4 g, 39.15 mmol, 4.88 mL, 1 eq) and Py (3.41 g, 43.06 mmol, 3.48 mL, 1.1 eq), DMAP (478.27 mg, 3.91 mmol, 0.1 eq) in DCM (80 mL) was added dropwise 2-bromoacetyl bromide (7.90 g, 39.15 mmol, 3.41 mL, 1 eq) in DCM (20 mL) at 0 °C under N2. After addition, the resulting mixture was stirred at 25 °C for 3 h. The reaction mixture was concentrated under reduced pressure to residue. The residue was purified by flash silica gel chromatography (40 g SepaFlash® Silica Flash Column, PE : EtOAc: 0~10%) to give compound hexyl 2-bromoacetate (34a, 4.2 g, 18.83 mmol, 48.1% yield) as yellow oil. ¹H NMR (400 MHz, CDCl3) δ = 4.18 (t, J = 6.8 Hz, 2H), 3.84 (s, 2H), 1.72 - 1.63 (m, 2H), 1.43 - 1.26 (m, 6H), 0.91 (t, J = 6.8 Hz, 3H). [00533] Step 2: bis(2-butyloctyl) 10-[(1-tert-butoxycarbonyl-4- piperidyl)methylamino]nonadecanedioate (34b)

[00534] A mixture of tert-butyl 4-(aminomethyl)piperidine-1-carboxylate (867.81 mg, 4.05 mmol, 1.1 eq) and NaBH(OAc)3 (1.17 g, 5.52 mmol, 1.5 eq), HOAc (331.59 mg, 5.52 mmol, 316.11 μL, 1.5 eq) in DCM (30 mL) was stirred at 25 °C for 1 h, and then bis(2-butyloctyl) 10- oxononadecanedioate (2.5 g, 3.68 mmol, 1 eq) was added. The resulting mixture was stirred at 25 °C for 15 h. The residue was diluted with DCM (50 mL) and the pH was adjusted to 11-12 with 4 M aqueous NaOH. The mixture was extracted with DCM (40 mL × 3) and the combined organic layers were washed with saturated brine (50 mL), dried over Na₂SO₄, filtered and concentrated under reduced pressure to give residue. The residue was purified by flash silica gel chromatography (20 g SepaFlash® Silica Flash Column, DCM : MeOH: 0~10%) to give compound bis(2-butyloctyl) 10-[(1-tert-butoxycarbonyl-4- piperidyl)methylamino]nonadecanedioate (34b, 2.95g, 3.36 mmol, 91.3% yield) as yellow oil. [00535] Step 3: bis(2-butyloctyl) 10-[(1-tert-butoxycarbonyl-4-piperidyl)methyl-(2-hexoxy-2- oxo-ethyl)amino]nonadecanedioate (34c)

[00536] To a solution of bis(2-butyloctyl) 10-[(1-tert-butoxycarbonyl-4- piperidyl)methylamino]nonadecanedioate (34b, 1.6 g, 1.82 mmol, 1 eq) and hexyl 2- bromoacetate (488.22 mg, 2.19 mmol, 1.2 eq) in MeCN (20 mL) were added K2CO3 (504.07 mg, 3.65 mmol, 2 eq) and NaI (27.33 mg, 182.36 μmol, 0.1 eq). The mixture was stirred at 60 °C for 16 h. The reaction mixture was filter and the filtrate was concentrated under reduced pressure to give compound bis(2-butyloctyl) 10-[(1-tert-butoxycarbonyl-4-piperidyl)methyl-(2- hexoxy-2-oxo-ethyl)amino]nonadecanedioate (34c, 1.85 g, crude) as yellow oil. The crude product was used for next step further purification. LCMS: [M+H]⁺: 1020.5 [00537] Step 4: bis(2-butyloctyl) 10-[(2-hexoxy-2-oxo-ethyl)-(4- piperidylmethyl)amino]nonadecanedioate (34d)

[00538] To a solution of bis(2-butyloctyl) 10-[(1-tert-butoxycarbonyl-4-piperidyl)methyl-(2- hexoxy-2-oxo-ethyl)amino]nonadecanedioate (34c, 1.8 g, 1.77 mmol, 1 eq) in DCM (15 mL) was added HCl/dioxane (2 M, 10.00 mL, 11.33 eq). The mixture was stirred at 25 °C for 2 h. The reaction mixture was concentrated under reduced pressure to give compound bis(2- butyloctyl) 10-[(2-hexoxy-2-oxo-ethyl)-(4-piperidylmethyl)amino]nonadecanedioate (34d, 1.7 g, crude, HCl) as yellow oil. LCMS: [M+H]⁺: 919.9 [00539] Step 5: bis(2-butyloctyl) 10-[(2-hexoxy-2-oxo-ethyl)-[(1-methyl-4- piperidyl)methyl]amino]nonadecanedioate (34)

[00540] A mixture of bis(2-butyloctyl) 10-[(2-hexoxy-2-oxo-ethyl)-(4- piperidylmethyl)amino]nonadecanedioate (1 g, 1.05 mmol, 1 eq, HCl), (HCHO)n (157.05 mg, 5.23 mmol, 144.08 μL, 5 eq), NaBH3CN (131.48 mg, 2.09 mmol, 2 eq), NaOAc (257.44 mg, 3.14 mmol, 3 eq) in MeOH (20 mL) was degassed and purged with N23 times, and then the mixture was stirred at 25 °C for 16 h under N₂ atmosphere. The reaction mixture was filtered and the filtrate was concentrated under reduced pressure to give residue. The residue was purified by prep-HPLC (column: CD27-PHS XP phenyl-Hexyl 150 × 25 × 7um; mobile phase: [water (FA) - ACN]; gradient: 60%-90% B over 15 min), and then purified by flash silica gel chromatography (12 g SepaFlash® Silica Flash Column, DCM : MeOH: 0~5%) to give bis(2- butyloctyl) 10-[(2-hexoxy-2-oxo-ethyl)-[(1-methyl-4-piperidyl)methyl]amino]nonadecanedioate (34) (235 mg, 251.71 μmol, 33.6% yield, 99.99% purity) as colorless gum. LCMS: [M+H]⁺: 933.8¹H NMR (400 MHz, CDCl3) δ = 4.06 (t, J = 6.8 Hz, 2H), 3.97 (d, J = 5.6 Hz, 4H), 3.32 - 3.25 (m, 2H), 3.19 (s, 2H), 2.62 (s, 3H), 2.48 - 2.44 (m, 3H), 2.42 - 2.37 (m, 1H), 2.30 (t, J = 7.6 Hz, 4H), 2.06 - 2.02 (m, 2H), 1.66 - 1.59 (m, 8H), 1.53 - 1.49 (m, 2H), 1.36 - 1.21 (m, 64H), 0.94 - 0.86 (m, 15H). Example 35: bis(2-butyloctyl) 10-(((1-(2-hydroxyethyl)piperidin-4- yl)methyl)(octylsulfinyl)amino)nonadecanedioate (35) [00541] Step 1: bis(2-butyloctyl) 10-(((1-(2-((tert-butyldimethylsilyl)oxy)ethyl)piperidin-4- yl)methyl)(octylsulfinyl)amino)nonadecanedioate (35a)

[00542] To a solution of bis(2-butyloctyl) 10-[[1-[2-[tert-butyl(dimethyl)silyl]oxyethyl]-4- piperidyl]methylamino]nonadecanedioate (15b) (1 g, 1.07 mmol, 1.0 eq) and TEA (757.07 mg, 7.48 mmol, 1.04 mL, 7.0 eq) in DCM (10 mL) was added octane-1-sulfinyl chloride (11a) (1.26 g, 6.41 mmol, 6.0 eq) in DCM (5 mL) at 0 °C. The mixture was stirred at 25 °C for 10 h. The reaction was concentrated and was purified by flash silica gel chromatography (ISCO®; 20 g SepaFlash® Silica Flash Column, Eluent of 0 ~ 5% MeOH / DCM gradient @ 35 mL / min) to give compound bis(2-butyloctyl) 10-[[1-[2-[tert-butyl(dimethyl)silyl]oxyethyl]-4- piperidyl]methyl-octylsulfinyl-amino]nonadecanedioate (35a) (1.15 g, 944.44 μmol, 88.4% yield, 90% purity) as a yellow oil. ¹H NMR (400 MHz, CDCl₃) δ = 4.11 - 4.00 (m, 2H), 3.98 - 3.93 (m, 4H), 3.74 - 3.57 (m, 2H), 3.25 - 3.02 (m, 4H), 3.01 - 2.90 (m, 2H), 2.84 - 2.71 (m, 3H), 2.58 - 2.48 (m, 1H), 2.32 - 2.26 (m, 4H), 2.00 - 1.90 (m, 2H), 1.66 - 1.57 (m, 8H), 1.46 - 1.38 (m, 4H), 1.33 - 1.22 (m, 64H), 0.91 - 0.85 (m, 24H), 0.16 - 0.03 (m, 6H). [00543] Step 2: bis(2-butyloctyl) 10-(((1-(2-hydroxyethyl)piperidin-4- yl)methyl)(octylsulfinyl)amino)nonadecanedioate (35)

[00544] A mixture of bis(2-butyloctyl) 10-[[1-[2-[tert-butyl(dimethyl)silyl]oxyethyl]-4- piperidyl]methyl-octylsulfinyl-amino]nonadecanedioate (35a) (600 mg, 547.50 μmol, 1.0 eq) in THF (10 mL), then tetrabutylammonium;fluoride;trihydrate (1.0 M, 821.26 μL, 1.5 eq) was added slowly at 0 °C, the mixture was degassed and purged with N₂3 times, and then the mixture was stirred at 25 °C for 10 h under N2. The reaction was concentrated. The residue was purified by flash silica gel chromatography (ISCO®; 12 g SepaFlash® Silica Flash Column, Eluent of 0 ~ 5% MeOH / DCM gradient @ 25 mL / min) to give compound bis(2-butyloctyl) 10-[[1-(2-hydroxyethyl)-4-piperidyl]methyl-octylsulfinyl-amino]nonadecanedioate (35) (130 mg, 130.37 μmol, 23.8% yield, 98.44% purity) as a yellow oil. LCMS: [M+H]⁺: 981.8¹H NMR (400 MHz, CD₃OD-d4) δ = 4.08 - 3.92 (m, 4H), 3.83 - 3.71 (m, 2H), 3.28 - 3.18 (m, 2H), 3.16 - 3.05 (m, 2H), 2.87 - 2.63 (m, 5H), 2.52 - 2.35 (m, 2H), 2.32 (t, J = 7.2 Hz, 4H), 1.94 - 1.75 (m, 3H), 1.67 - 1.59 (m, 8H), 1.58 - 1.53 (m, 2H), 1.52 - 1.43 (m, 4H), 1.40 - 1.26 (m, 62H), 0.96 - 0.87 (m, 15H). Example 36: 9-(((1-methylpiperidin-4-yl)methyl)(octylsulfinyl)amino)heptadecane-1,17-diyl bis(3-butylnonanoate) (36) [00545] Step 1: methyl 3-butylnonanoate (36a):

[00546] To a solution of CuBr (968.98 mg, 6.75 mmol, 205.73 μL, 0.1 eq) in THF (130 mL) was added LiCl (572.68 mg, 13.51 mmol, 276.92 μL, 0.2 eq). The solution was stirred at 0 °C for 10 min under N2. compound methyl (E)-non-2-enoate (11.5 g, 67.55 mmol, 1.0 eq) was added into the solution. TMSCl (8.07 g, 74.30 mmol, 9.43 mL, 1.1 eq) was dropwise into the solution. The solution was stirred at 0 °C for 15 min. Compound bromo(butyl)magnesium (1.0 M, 81.06 mL, 1.2 eq) was dropwise into the solution. The solution was stirred at 0 °C for 2 h under N2. The reaction mixture was poured into the NH4Cl (aq, sat, 80 mL), and the solution was extracted with EtOAc (150 mL). The organic layer was washed with brine (100 mL).The organic layer was dried over sodium sulfate, filtered and concentrated under reduced pressure to give a residue. The residue was purified by flash silica gel chromatography (ISCO®; 220 g SepaFlash® Silica Flash Column, Eluent of 0 ~ 1% EtOAc / PE gradient @ 100 mL / min) to give compound methyl 3-butylnonanoate (36a) (11.18 g, 44.06 mmol, 65.2% yield, 90% purity) as colorless oil. ¹H NMR (400 MHz, CDCl3) δ = 3.74 - 3.58 (m, 3H), 2.23 (d, J = 6.8 Hz, 2H), 1.88 - 1.78 (m, 1H), 1.32 - 1.21 (m, 16H), 0.93 - 0.85 (m, 6H). [00547] Step 2: 3-butylnonanoic acid (36b)

[00548] A mixture of methyl 3-butylnonanoate (36a) (6 g, 26.27 mmol, 1.0 eq) and NaOH (2.10 g, 52.55 mmol, 2.0 eq) in THF (60 mL) and H2O (10 mL) was degassed and purged with N₂3 times, and then the mixture was stirred at 60 °C for 5 h under N₂. The reaction mixture cooled down to room temperature and concentrated , the diluted with EtOAc (60 mL), pH adjust to pH = 4 by addition 1N HCl, and extracted with EtOAc 60 mL (30 mL × 2). The combined organic layers were washed with brine (50 mL), dried over anhydrous sodium sulfate, filtered and concentrated under reduced pressure to give a residue. The residue was purified by flash silica gel chromatography (ISCO®; 120 g SepaFlash® Silica Flash Column, Eluent of 0~10% EtOAc / PE gradient @ 100 mL / min) to give compound 3-butylnonanoic acid (36b) (3.02 g, 14.09 mmol, 53.6% yield) as a yellow oil.¹H NMR (400 MHz, CDCl₃) δ = 2.29 (d, J = 6.8 Hz, 2H), 1.94 - 1.79 (m, 1H), 1.40 - 1.24 (m, 16H), 1.00 - 0.83 (m, 6H). [00549] Step 3: diethyl 9-isocyano-9-tosylheptadecanedioate (36c)

[00550] To a 500 mL three neck round bottom flask under N2 was added DMSO (200 mL), followed by the addition of NaH (18.95 g, 473.78 mmol, 60% purity, 2.5 eq) in portions over a period of 0.5 h. The mixture was allowed to stir for 0.5 h at 25 °C following the addition, then 1- (isocyanomethylsulfonyl)-4-methyl-benzene (37 g, 189.51 mmol, 1.0 eq) was added in portions over 0.5 h, followed by the addition of TBAI (7.00 g, 18.95 mmol, 0.1 eq) in one portion. The resulting mixture was stirred for 0.25 h at 25 °C, then ethyl 8-bromooctanoat (99.96 g, 397.98 mmol, 2.1 eq) was added over a period of 0.5 h. The mixture was stirred at 25 °C for 1 h. The reaction mixture was quenched by addition of water (200 mL) at 0 °C, and then extracted with PE (150 mL × 2). The combined organic layers were washed with brine (200 mL), dried over anhydrous sodium sulfate, filtered and concentrated under reduced pressure to give compound diethyl 9-cyano-9-(p-tolylsulfonyl)heptadecanedioate (36c) (77 g, crude) as a brown oil. [00551] Step 4: diethyl 9-oxoheptadecanedioate (36d)

[00552] To a 250 mL three neck round bottom flask was added DCM (500 mL), followed by the addition of diethyl 9-isocyano-9-(p-tolylsulfonyl)heptadecanedioate (36c) (77 g, 143.73 mmol, 1.0 eq) under N₂, HCl (12 M, 94.77 mL, 7.91 eq) was added in portions over a period of 1 h. The two-phase mixture was stirred for 12 h at 25 °C. The reaction mixture was quenched by addition water (150 mL) at 0 °C and then extracted with PE (150 mL × 2). The combined organic layers were washed with brine (200 mL), dried over anhydrous sodium sulfate, filtered and concentrated under reduced pressure to give a residue. The residue was purified by column chromatography (SiO2, PE / EtOAc = 1 / 0 to 20 / 1) to give compound diethyl 9- oxoheptadecanedioate (36d) (23 g, 55.87 mmol, 38.9% yield, 90% purity) as a yellow oil. ¹H NMR (400 MHz, CDCl₃) δ = 4.14 - 4.08 (m, 4H), 2.39 - 2.34 (m, 4H), 2.29 - 2.25 (m, 4H), 1.66 - 1.48 (m, 10H), 1.31 - 1.22 (m, 20H).

[00554] To a 250 mL three neck round bottom flask was added EtOH (180 mL) and H2O (60 mL), followed by the addition of diethyl 9-oxoheptadecanedioate (22 g, 59.38 mmol, 1.0 eq) under N₂, NaOH (7.12 g, 178.13 mmol, 3.0 eq) was added in portions over a period of 1 h. The two-phase mixture was stirred for 12 h at 20 °C. The reaction mixture was concentrated and diluted with H2O (100 mL) and then extracted with EtOAc (150 mL × 2). the aqueous phase with 1N HCl adjust to pH = 3, then filtered and the filtered cake was concentrated under reduced pressure to give compound 9-oxoheptadecanedioic acid (36e) (18.2 g, 57.89 mmol, 97.49% yield) as a white solid.¹H NMR (400 MHz, DMSO-d6) δ = 2.44 - 2.32 (m, 4H), 2.23 - 2.09 (m, 4H), 1.61 - 1.35 (m, 8H), 1.35 - 1.01 (m, 12H). [00555] Step 6: dimethyl 9-oxoheptadecanedioate (36f)

[00556] A mixture of 9-oxoheptadecanedioic acid (36e) (10 g, 31.81 mmol, 1.0 eq) in MeOH (120 mL) was degassed and purged with N23 times, then H2SO4 (15.60 g, 159.03 mmol, 8.48 mL, 5.0 eq) was added at 0 °C and then the mixture was stirred at 70 °C for 10 h under N2. The reaction mixture was quenched by addition H₂O (100 mL) at 0 °C, and then diluted with H₂O (150 mL) and extracted with EtOAc 200 mL (100 mL × 2). The combined organic layers were washed with brine 200 mL (100 mL × 2), dried over anhydrous sodium sulfate, filtered and concentrated under reduced pressure to give a residue. The residue was purified by flash silica gel chromatography (ISCO®; 120g SepaFlash® Silica Flash Column, Eluent of 0 ~ 15% EtOAc / PE gradient @ 80 mL / min) to give compound dimethyl 9-oxoheptadecanedioate (36f) (7.67 g, 21.28 mmol, 66.9% yield, 95% purity) as a white solid. ¹H NMR (400 MHz, CDCl₃) δ = 3.66 (s, 6H), 2.37 (t, J = 7.2 Hz, 4H), 2.30 (t, J = 7.6 Hz, 4H), 1.65 - 1.53 (m, 8H), 1.33 - 1.24 (m, 12H). [00557] Step 7: dimethyl 9-(((1-methylpiperidin-4-yl)methyl)amino)heptadecanedioate (36g)

[00558] A mixture of (1-methyl-4-piperidyl)methanamine (2.8 g, 21.84 mmol, 1.0 eq), NaBH(OAc)3 (6.94 g, 32.76 mmol, 1.5 eq), HOAc (1.97 g, 32.76 mmol, 1.88 mL, 1.5 eq) in DCM (50 mL) was degassed and purged with N23 times, dimethyl 9-oxoheptadecanedioate (36f) (8.23 g, 24.02 mmol, 1.1 eq) in DCM (30 mL) was added and then the mixture was stirred at 25 °C for 10 h under N2. The mixture pH was adjusted to 9 - 10 was with 1 N NaOH (20 mL), the aqueous phase was extracted with EtOAc (35 mL × 3). The combined organic phase was washed with brine (40 mL), dried with anhydrous sodium sulfate, filtered and concentrated in vacuum to give crude product. The residue was purified by flash silica gel chromatography (ISCO®; 120 g SepaFlash® Silica Flash Column, Eluent of 0 ~ 5% MeOH / DCM @ 80 mL / min) to give compound dimethyl 9-[(1-methyl-4-piperidyl)methylamino]heptadecanedioate (36g) (8.19 g, 18.01 mmol, 82.5% yield) as a yellow oil.¹H NMR (400 MHz, CDCl3) δ = 3.66 (s, 6H), 2.88 - 2.78 (m, 2H), 2.44 - 2.35 (m, 3H), 2.33 - 2.26 (m, 4H), 2.25 (s, 3H), 1.94 - 1.86 (m, 2H), 1.73 - 1.68 (m, 2H), 1.64 - 1.54 (m, 4H), 1.40 - 1.17 (m, 24H). [00559] Step 8: dimethyl 9-(((benzyloxy)carbonyl)((1-methylpiperidin-4- yl)methyl)amino)heptadecanedioate (36h)

[00560] A mixture of dimethyl 9-[(1-methyl-4-piperidyl)methylamino]heptadecanedioate (36g) (2.5 g, 5.50 mmol, 1.0 eq), DIEA (1.42 g, 11.00 mmol, 1.92 mL, 2.0 eq) in DCM (25 mL) was degassed and purged with N₂3 times, then benzyl (2,5-dioxopyrrolidin-1-yl) carbonate (1.64 g, 6.60 mmol, 1.2 eq) was added at 0 °C, and the reaction was stirred at 25 °C for 3 h. The reaction mixture was concentrated under reduced pressure to give a residue. The residue was purified by flash silica gel chromatography (ISCO®; 80 g SepaFlash® Silica Flash Column, Eluent of 0 ~ 5% MeOH / DCM gradient @ 100 mL / min) to give compound dimethyl 9- [benzyloxycarbonyl-[(1-methyl-4-piperidyl)methyl]amino]heptadecanedioate (36h) (3.9 g, 5.30 mmol, 96.4% yield, 80% purity) as a yellow oil. ¹H NMR (400 MHz, CDCl₃) δ = 7.43 - 7.20 (m, 5H), 5.25 - 4.98 (m, 2H), 3.65 (s, 6H), 3.33 - 3.07 (m, 2H), 3.07 - 2.86 (m, 2H), 2.62 - 2.57 (m, 4H), 2.56 - 2.52 (m, 1H), 2.49 - 2.36 (m, 2H), 2.31 - 2.22 (m, 4H), 1.84 - 1.64 (m, 2H), 1.64- 1.50 (m, 6H), 1.50 - 1.34 (m, 4H), 1.31 - 1.09 (m, 16H). [00561] Step 9: benzyl (1,17-dihydroxyheptadecan-9-yl)((1-methylpiperidin-4- yl)methyl)carbamate (36i)

[00562] A mixture of dimethyl 9-[benzyloxycarbonyl-[(1-methyl-4- piperidyl)methyl]amino]heptadecanedioate (36h) (3.9 g, 6.62 mmol, 1.0 eq) in THF (80 mL) was degassed and purged with N23 times and cooled down to 0 °C, then LiAlH4 (2.5 M, 5.56 mL, 2.1 eq) was added via injection at 0 °C, and then the mixture was stirred at 25 °C for 1 h under N₂. After completion, the reaction mixture was diluted with THF (20 mL), then successively was added H₂O (0.53 mL), aq.NaOH (0.53 mL, 15%), H₂O (1.6 mL) and dried over anhydrous sodium sulfate at 0 °C under N2. The reaction mixture was filtered and the filtrate was concentrated in vacuum to give residue. The residue was purified by flash silica gel chromatography (ISCO®; 40g SepaFlash® Silica Flash Column, Eluent of 0 ~ 5% MeOH / DCM gradient @ 50 mL / min) to give compound benzyl N-[9-hydroxy-1-(8- hydroxyoctyl)nonyl]-N-[(1-methyl-4-piperidyl)methyl]carbamate (36i) (2.49 g, 4.44 mmol, 67.0% yield, 95% purity) as a colorless oil. ¹H NMR (400 MHz, CDCl₃) δ = 7.44 - 7.20 (m, 5H), 5.11 (s, 2H), 3.88 - 3.69 (m, 1H), 3.62 (t, J = 6.8 Hz, 4H), 3.09 - 2.94 (m, 2H), 2.93 - 2.72 (m, 2H), 2.23 (s, 3H), 1.97 - 1.72 (m, 5H), 1.68 - 1.43 (m, 10H), 1.37 - 1.11 (m, 22H). [00563] Step 10: 9-(((benzyloxy)carbonyl)((1-methylpiperidin-4- yl)methyl)amino)heptadecane-1,17-diyl bis(3-butylnonanoate) (36j)

[00564] To a solution of benzyl N-[9-hydroxy-1-(8-hydroxyoctyl)nonyl]-N-[(1-methyl-4- piperidyl)methyl]carbamate (36i) (1.2 g, 2.25 mmol, 1.0 eq), 3-butylnonanoic acid (36b) (1.21 g, 5.63 mmol, 2.5 eq), TEA (1.14 g, 11.26 mmol, 1.57 mL, 5.0 eq) and DMAP (825.46 mg, 6.76 mmol, 3.0 eq) in DCM (30 mL) was added EDCI (1.08 g, 5.63 mmol, 2.5 eq). The mixture was stirred at 25 °C for 10 h. The reaction was concentrated. The residue was purified by flash silica gel chromatography (ISCO®; 20 g SepaFlash® Silica Flash Column, Eluent of 0 ~ 5% MeOH / DCM gradient @ 50 mL / min) to give compound [9-[benzyloxycarbonyl-[(1-methyl-4- piperidyl)methyl]amino]-17-(3-butylnonanoyloxy)heptadecyl] 3-butylnonanoate (36j) (1.53 g, 1.52 mmol, 67.5% yield, 92% purity) as a yellow oil. ¹H NMR (400 MHz, CDCl₃) δ = 7.42 - 7.26 (m, 5H), 5.11 (s, 2H), 4.16 - 3.95 (m, 4H), 3.07 - 2.93 (m, 2H), 2.92 - 2.77 (m, 2H), 2.33 - 2.16 (m, 7H), 1.96 - 1.79 (m, 4H), 1.67 - 1.56 (m, 6H), 1.55 - 1.41 (m, 4H), 1.37 - 1.12(m, 56H), 1.02 - 0.78 (m, 12H). [00565] Step 11: 9-(((1-methylpiperidin-4-yl)methyl)amino)heptadecane-1,17-diyl bis(3- butylnonanoate) (36k)

[00566] To a solution of [9-[benzyloxycarbonyl-[(1-methyl-4-piperidyl)methyl]amino]-17-(3- butylnonanoyloxy)heptadecyl] 3-butylnonanoate (36j) (1.53 g, 1.65 mmol, 1.0 eq) in MeOH (15 mL) and THF (15 mL) was added Pd/C (10%, 200 mg), Pd(OH)₂/C (20%, 200 mg) under N₂. The suspension was degassed and purged with H23 times. The mixture was stirred under H2 (20 Psi ) at 40 °C for 3 h. The reaction mixture was filtered with DCM (50 mL) and MeOH (30 mL) and the filtrate concentrated under reduced pressure to give a residue. The residue was purified by flash silica gel chromatography (ISCO®; 40 g SepaFlash® Silica Flash Column, Eluent of 0 ~ 5% MeOH / DCM gradient @ 50 mL / min) to give compound [17-(3-butylnonanoyloxy)-9- [(1-methyl-4-piperidyl)methylamino]heptadecyl]3-butylnonanoate (36k) (755 mg, 906.40 μmol, 54.8% yield, 95% purity) as a yellow oil.¹H NMR (400 MHz, CDCl₃) δ = 4.06 (t, J = 6.8 Hz, 4H), 2.91 - 2.79 (m, 2H), 2.48 - 2.37 (m, 3H), 2.27 (s, 3H), 2.23 (d, J = 6.8 Hz, 4H), 1.97 - 1.87 (m, 2H), 1.87 - 1.80 (m, 2H), 1.76 - 1.71 (m, 2H), 1.66 - 1.57 (m, 6H), 1.37 - 1.19 (m, 58H), 1.01 - 0.79 (m, 12H). [00567] Step 12: 9-(((1-methylpiperidin-4-yl)methyl)(octylsulfinyl)amino)heptadecane-1,17- diyl bis(3-butylnonanoate) (36)

[00568] To a solution of [17-(3-butylnonanoyloxy)-9-[(1-methyl-4- piperidyl)methylamino]heptadecyl] 3-butylnonanoate (36k) (710 mg, 897.23 μmol, 1.0 eq), DMAP (21.92 mg, 179.45 μmol, 0.2 eq) and TEA (272.37 mg, 2.69 mmol, 374.65 μL, 3.0 eq) in DCM (10 mL) was added octane-1-sulfinyl chloride (1.06 g, 5.38 mmol, 6.0 eq) in DCM (2 mL) at 0 °C. The mixture was stirred at 25 °C for 2 h. The reaction was concentrated. The residue was purified by flash silica gel chromatography (ISCO®; 20 g SepaFlash® Silica Flash Column, Eluent of 0 ~ 5% MeOH / DCM gradient @ 25 mL / min) then was further purified by prep- TLC (SiO2, DCM : MeOH = 10 : 1, 0.5%NH3•H2O) to give compound [17-(3- butylnonanoyloxy)-9-[(1-methyl-4-piperidyl)methyl-octylsulfinyl-amino]heptadecyl] 3- butylnonanoate (36) (70.1 mg, 73.25 μmol, 25.5% yield, 99.44% purity) was obtained as a yellow oil. LCMS: [M+H]⁺: 951.4¹H NMR (400 MHz, CDCl3) δ = 4.13 - 4.00 (m, 4H), 3.14 - 3.00 (m, 1H), 3.00 - 2.79 (m, 3H), 2.78 - 2.61 (m, 2H), 2.60 - 2.48 (m, 1H), 2.37 - 2.15 (m, 7H), 1.98 - 1.86 (m, 2H), 1.81 - 1.68 (m, 5H), 1.65 - 1.58 (m, 6H), 1.49 - 1.39 (m, 4H), 1.37 - 1.20 (m, 64H), 0.92 - 0.85 (m, 15H). Example 37: bis(2-butyloctyl) 10-(((1-methylpiperidin-4- yl)methyl)(octyloxy)amino)nonadecanedioate (37) [00569] Step 1: 2-(octyloxy)isoindoline-1,3-dione (37a)

[00570] To a solution of 1-bromooctane (10 g, 51.78 mmol, 9.01 mL, 1.0 eq) and 2- hydroxyisoindoline-1,3-dione (10.14 g, 62.14 mmol, 1.2 eq) in DMF (200 mL), K2CO3 (8.59 g, 62.14 mmol, 1.2 eq) was added in two portions. The reaction mixture was allowed to 12 h at 25 °C. The solvent was evaporated under reduced pressure. The resulting solid was dissolved in EtOAc (100 mL) and washed with H2O (100 mL). After separation, the organic layer was washed with H₂O (2 × 30 mL) and with brine (2 × 30 mL). The organic layer was dried over anhydrous sodium sulfate and concentrated under pressure to give compound 2- octoxyisoindoline-1,3-dione (37a) (15 g, crude) as a yellow oil. ¹H NMR (400 MHz, CDCl3) δ = 7.87 - 7.77 (m, 2H), 7.77 - 7.67 (m, 2H), 4.26 - 4.12 (m, 2H), 1.81 - 1.71 (m, 2H), 1.53 - 1.39 (m, 2H), 1.34 - 1.24 (m, 8H), 0.89 - 0.83 (m, 3H). [00571] Step 2: O-octylhydroxylamine (37b)

[00572] To a solution of 2-octoxyisoindoline-1,3-dione (37a) (15 g, 54.48 mmol, 1.0 eq) in EtOH (400 mL) and DCM (400 mL), NH2NH2•H2O (8.18 g, 163.43 mmol, 7.93 mL, 100% purity, 3.0 eq) was added drop wise at 25 °C. The reaction was stirred at 25 °C for 48 h. The reaction was concentrated, then the residue was dissolved in DCM (300 mL) and was quenched with 1 N NaOH (30 mL). The organic layer was washed with H2O 100 mL (2 × 50 mL), brine 100 mL (2 × 50 mL), dried over anhydrous sodium sulfate and concentrated under reduced pressure. The residue was purified by column chromatography (SiO₂, PE / EtOAc = 1 / 0 to 10 / 1) to give compound O-octylhydroxylamine (37b) (2.4 g, 14.05 mmol, 25.8% yield, 85% purity) as a colorless oil. ¹H NMR (400 MHz, CDCl3) δ = 5.80 - 4.59 (m, 2H), 3.71 - 3.58 (m, 2H), 1.62-1.49 (m, 2H), 1.33 - 1.19 (m, 10H), 0.86 (t, J = 6.8 Hz, 3H). [00573] Step 3: bis(2-butyloctyl) 10-((octyloxy)imino)nonadecanedioate (37c)

[00574] A mixture of O-octylhydroxylamine (37b) (962.43 mg, 6.63 mmol, 1.5 eq), bis(2- butyloctyl)10-oxononadecanedioate (1f) (3 g, 4.42 mmol, 1.0 eq), HOAc (530.57 mg, 8.84 mmol, 505.78 μL, 2.0 eq) and NaBH3CN (832.83 mg, 13.25 mmol, 3.0 eq) in MeOH (40 mL)and DCE (8 mL) was degassed and purged with N₂3 times, the reaction was stirred 50 °C for 16 h under N₂. The mixture was concentrated, and pH was adjusted to 9 - 10 was with 1 N NaOH (10 mL), the aqueous phase was extracted with DCM 50 mL (25 mL × 2). The combined organic phase was washed with brine (60 mL), dried with anhydrous sodium sulfate, filtered and concentrated in vacuum to give crude product. The residue was purified by flash silica gel chromatography (ISCO®; 40g SepaFlash® Silica Flash Column, Eluent of 0 ~ 5% MeOH / DCM @ 60 mL / min) to give compound bis(2-butyloctyl) 10-octoxyiminononadecanedioate (37c) (860 mg, 959.90 μmol, 21.7% yield, 90% purity) as a yellow oil. ¹H NMR (400 MHz, CDCl3) δ = 4.03 - 3.92 (m, 6H), 2.34 - 2.22 (m, 6H), 2.16 - 2.08 (m, 2H), 1.64 -1.60 (m 8H), 1.52 - 1.43 (m, 4H), 1.34 - 1.21 (m, 58H), 0.95 - 0.83 (m, 15H). [00575] p 4: 1-methylpiperidine-4-carbaldehyde (37d)

[00576] e oxalylchloride (9.82 g, 77.40 mmol, 6.78 mL, 2.0 eq) was dissolved in anhydrous DCM (60 mL) under N₂ and cooled to -70 °C, DMSO (6.65 g, 85.14 mmol, 6.65 mL, 2.2 eq) in DCM (10 mL) was added dropwise to the reaction solution and stirred at -70 °C for 15 min, then (1-methyl-4-piperidyl)methanol (5 g, 38.70 mmol, 1.0 eq) in DCM (15 mL) was added dropwise to the reaction solution, and the reaction solution was stirred at -70 °C for 1 h. TEA (19.58 g, 193.50 mmol, 26.93 mL, 5.0 eq) was added to the reaction solution, and the reaction solution was slowly warmed up to 25 °C and stirred for 15 minutes. Add saturated Na2CO3 aqueous solution (30 mL) to quench, extract with DCM (70 mL × 2), wash with brine (70 mL × 2), dried with anhydrous sodium sulfate, filtered and concentrated to give a residue. The residue was purified by flash silica gel chromatography (ISCO®; 120 g SepaFlash® Silica Flash Column, Eluent of 0 ~ 20% MeOH / DCM gradient @ 100 mL / min) to give compound 1- methylpiperidine-4-carbaldehyde (37d) (1.77 g, 13.92 mmol, 36.0% yield) as a brown oil. ¹H NMR (400 MHz, CDCl₃) δ = 9.62 (s, 1H), 2.82 - 2.72 (m, 2H), 2.27 (s, 3H), 2.25 - 2.19 (m, 1H), 2.14 - 2.04 (m, 2H), 1.96 - 1.88 (m, 2H), 1.76 - 1.67 (m, 2H). [00577] Step 5: bis(2-butyloctyl) 10-(((1-methylpiperidin-4- yl)methyl)(octyloxy)amino)nonadecanedioate (37)

[00578] To a clear solution of bis(2-butyloctyl) 10-(octoxyamino)nonadecanedioate (37c) (500 mg, 618.55 μmol, 1.0 eq) in HOAc (8 mL) was added NaBH₃CN (38.87 mg, 618.55 μmol, 1.0 eq) in single portion and stirred at 25 °C for 1 h. 1-methylpiperidine-4-carbaldehyde (786.69 mg, 6.19 mmol, 10.0 eq) was added to the reaction mixture and stirred further for 1 h at 25 °C. Finally, second portion of NaBH₃CN (38.87 mg, 618.55 μmol, 1.0 eq) was added to the resultant turbid mixture and stirred for 2 h. The mixture pH was adjusted to 9 - 10 was with 1 N NaOH (10 mL), the aqueous phase was extracted with DCM (15 mL × 3). The combined organic phase was washed with brine (10 mL), dried with anhydrous sodium sulfate, filtered and concentrated in vacuum to give crude product. The residue was purified by flash silica gel chromatography (ISCO®; 20 g SepaFlash® Silica Flash Column, Eluent of 0 ~ 4% MeOH / DCM @ 30 mL / min) to give compound bis(2-butyloctyl) 10-[(1-methyl-4-piperidyl)methyl-octoxy- amino]nonadecanedioate (37) (207.31 mg, 225.43 μmol, 36.5% yield, 99.99% purity) was obtained as a yellow oil. LCMS: [M+H]⁺: 919.7¹H NMR (400 MHz, CDCl3) δ = 3.98 (d, J = 6.0 Hz, 4H), 3.59 (t, J = 6.8 Hz, 2H), 2.92 - 2.81 (m, 2H), 2.70 - 2.58 (m, 1H), 2.48 - 2.39 (m, 2H), 2.36 - 2.21 (m, 7H), 1.99 - 1.90 (m, 2H), 1.89 - 1.79 (m, 3H), 1.66 - 1.57 (m, 6H), 1.52 - 1.45 (m, 4H), 1.37-1.20 (m, 66H), 0.96 - 0.83 (m, 15H). Example 38: [17-(3-butylnonanoyloxy)-9-[(1-methyl-4-piperidyl)methyl- (octylsulfonimidoyl)amino]heptadecyl] 3-butylnonanoate (38) [00579] Step 1: [17-(3-butylnonanoyloxy)-9-[(1-methyl-4-piperidyl)methyl- (octylsulfonimidoyl)amino]heptadecyl] 3-butylnonanoate (38)

[00580] A mixture of [17-(3-butylnonanoyloxy)-9-[(1-methyl-4-piperidyl)methyl- octylsulfinyl-amino]heptadecyl] 3-butylnonanoate (36) (160 mg, 156.37 μmol, 1.0 eq), PhI(OAc)2 (151.10 mg, 469.11 μmol, 3.0 eq) and ammonium carbamic acid (48.83 mg, 625.48 μmol, 4.0 eq) in MeOH (3 mL) was degassed and purged with N₂ for 3 times, and then the mixture was stirred at 20 °C for 2 h under N₂. The reaction was concentrated and was purified by prep-HPLC (FA condition; column: CD12-WePure Biotech Phenyl-Hexyl 150 × 25 mm × 7 um; mobile phase: [H₂O (0.225% FA) - ACN]; gradient: 50% - 80% B over 15.0 min) to give compound [17-(3-butylnonanoyloxy)-9-[(1-methyl-4-piperidyl)methyl- (octylsulfonimidoyl)amino]heptadecyl] 3-butylnonanoate (38) (70.11 mg, 46.4% yield, 99.99% purity) as a yellow oil. LCMS: [M+H]⁺: 966.8. ¹H NMR (400 MHz, CDCl3) δ = 4.19 - 4.14 (m, 1H), 4.11 - 4.01 (m, 4H), 3.69 (s, 3H), 3.15 - 3.05 (m, 2H), 2.99 - 2.90 (m, 1H), 2.89 - 2.73 (m, 2H), 2.59 - 2.47 (m, 1H), 2.22 (d, J = 6.8 Hz, 4H), 2.16 - 1.99 (m, 2H), 1.88 - 1.73 (m, 5H), 1.66 - 1.57 (m, 6H), 1.50 - 1.41 (m, 4H), 1.38 - 1.17 (m, 64H), 0.91 - 0.85 (m, 15H). Example 39: 9-(N-((1-methylpiperidin-4-yl)methyl)octylsulfonamido)heptadecane-1,17-diyl bis(3-butylnonanoate) (39)

[00581] To a solution of [17-(3-butylnonanoyloxy)-9-[(1-methyl-4-piperidyl)methyl- octylsulfinyl-amino]heptadecyl] 3-butylnonanoate (36) (130 mg, 136.61 μmol, 1.0 eq) in DCM (0.5 mL), MeCN (0.5 mL) and H2O (0.8 mL), NaIO4 (35.06 mg, 163.94 μmol, 9.08 μL, 1.2 eq) and RuCl₃ (5.67 mg, 27.32 μmol, 1.82 μL, 0.2 eq) was added slowly at 0 °C and stirred for 30 min under N2. after completely, DCM (5 mL) was added and separated water phase, and exacted with DCM (8 mL × 3), washed with brine (15 mL), dried over anhydrous sodium sulfate, filtered and concentrated under reduced pressure to give a residue. The residue was purified by prep-HPLC (FA condition; column: CD12-WePure Biotech Phenyl-Hexyl 150 × 25 mm × 7 um; mobile phase: [H2O (0.225% FA) - ACN]; gradient: 55% - 85% B over 10.0 min)to give compound [17-(3-butylnonanoyloxy)-9-[(1-methyl-4-piperidyl)methyl-octylsulfonyl- amino]heptadecyl] 3-butylnonanoate (39) (61.29 mg, 63.34 μmol, 46.4% yield, 99.99% purity) as a yellow oil. LCMS: [M+H]⁺: 968.3. ¹H NMR (400 MHz, CDCl3) δ = 4.06 (t, J = 6.8 Hz, 4H), 3.53 - 3.27 (m, 3H), 3.05 - 2.92 (m, 2H), 2.90 - 2.81 (m, 2H), 2.68 (s, 3H), 2.64 - 2.55 (m, 1H), 2.23 (d, J = 6.8 Hz, 4H), 2.03 - 1.95 (m, 3H), 1.88 - 1.76 (m, 7H), 1.65 - 1.59 (m, 4H), 1.53 - 1.46 (m, 2H), 1.45 - 1.38 (m, 4H), 1.36 - 1.23 (m, 60H), 0.96 - 0.81 (m, 15H). Example 40: 9-((2,2-difluorononyl)((1-methylpiperidin-4-yl)methyl)amino)heptadecane-1,17- diyl bis(3-butylnonanoate) (40) [00582] Step 1: 2,2-difluorononane-1,1-diol (40a)

[00583] To a round-bottom flask equipped with a magnetic stir bar and charged with pyrrolidine-2-carboxylic acid (1.21 g, 10.55 mmol, 0.3 eq) and NFSI (55.42 g, 175.76 mmol, 5.0 eq) was added THF (130 mL). The mixture was stirred at 20 °C until all solids were dissolved and the nonanal (5 g, 35.15 mmol, 1.0 eq) dissolved in THF (30 mL) was then slowly added to the reaction mixture and the mixture was stirred for 16 h. The reaction diluted with MTBE (100 mL) and cooled down to -78 °C, filtered through a pad of silica gel, eluting with cold MTBE (100 mL × 6), the resultant organic layer was washed with saturated NaHCO3 (30 mL), brine (50 mL), dried over anhydrous sodium sulfate, filtered and concentrated. The residue was purified by flash silica gel chromatography (ISCO®; 120 g SepaFlash® Silica Flash Column, Eluent of 0 ~ 15% EtOAc / PE gradient @ 50 mL / min) to give compound 2,2-difluorononane- 1,1-diol (40a) (3.55 g, 18.09 mmol, 51.5% yield) as a yellow oil. ¹H NMR (400 MHz, DMSO- d6) δ = 4.84 - 4.48 (m, 1H), 1.96 - 1.73 (m, 2H), 1.49 - 1.36 (m, 2H), 1.33 - 1.17 (m, 8H), 0.84 (t, J = 6.4 Hz, 3H). [00584] Step 2: 2,2-difluorononanoic acid (40b) 2-methylbut-2-ene (7.0 eq) NaClO (20 e )

40a 40b [00585] To a solution of 2,2-difluorononane-1,1-diol (40a) (2.8 g, 14.27 mmol, 1.0 eq) in t- BuOH (80 mL) were added 2-methylbut-2-ene (7.00 g, 99.88 mmol, 10.58 mL, 7.0 eq), a solution of NaClO₂ (2.58 g, 28.54 mmol, 2.0 eq) and NaH₂PO₄ (5.14 g, 42.81 mmol, 3.0 eq) in H2O (40 mL). The latter solution is added to the former solution and the mixture was stirred at 25 °C for 2 h. the mixture was quenched with 1 N HCl adjust pH to 3 - 4 and extracted with EtOAc (15 mL× 3). The organic phase was washed with brine (20 mL× 3) and dried over anhydrous sodium sulfate. filtered and concentrated. The residue was purified by flash silica gel chromatography (ISCO®; 40 g SepaFlash® Silica Flash Column, Eluent of 0 ~ 8% MeOH / DCM gradient @60 mL/min) to give compound 2,2-difluorononanoic acid (40b) (1.5 g, 6.18 mmol, 43.3% yield, 80% purity) as a colorless oil. ¹H NMR (400 MHz, DMSO-d₆) δ = 2.21 - 2.11 (m, 2H), 1.50 - 1.32 (m, 10H), 0.96 (t, J = 6.8 Hz, 3H). [00586] Step 3: 2,2-difluorononanoyl chloride (40c)

[00587] A mixture of 2,2-difluorononanoic acid (40b) (1.5 g, 7.72 mmol, 1.0 eq) in DCM (20 mL) was degassed and purged with N₂ for 3 times, then SOCl₂ (1.38 g, 11.58 mmol, 841.43 μL, 1.5 eq) and DMF (56.45 mg, 772.33 μmol, 59.42 μL, 0.1 eq) were added slowly and then the mixture was stirred at 40 °C for 1h under N2. The reaction was concentrated under (35 °C) reduce pressure to give compound 2,2-difluorononanoyl chloride (40c) (1.6 g, crude) as a colorless oil. [00588] Step 4: dimethyl 9-(2,2-difluoro-N-((1-methylpiperidin-4- yl)methyl)nonanamido)heptadecanedioate (40d)

[00589] To a solution of dimethyl 9-[(1-methyl-4-piperidyl)methylamino]heptadecanedioate (1.35 g, 2.97 mmol, 1.0 eq), TEA (1.50 g, 14.85 mmol, 2.07 mL, 5.0 eq) in DCM (30 mL) was added 2,2-difluorononanoyl chloride (40c) (1.58 g, 7.42 mmol, 2.5 eq). The mixture was stirred at 25 °C for 15 min. The reaction mixture was partitioned between DCM (10 mL × 3) and H2O (20 mL). The organic phase was separated, washed with brine (10 mL × 2), dried over anhydrous sodium sulfate, filtered and concentrated under reduced pressure to give a residue. The residue was purified by flash silica gel chromatography (ISCO®; 40 g SepaFlash® Silica Flash Column, Eluent of 0 ~ 5% MeOH / DCM gradient @ 60 mL / min) to give compound dimethyl 9-[2,2-difluorononanoyl-[(1-methyl-4-piperidyl)methyl]amino]heptadecanedioate (40d) (1.69 g, 2.28 mmol, 76.7% yield, 85% purity) as a yellow oil. ¹H NMR (400 MHz, CDCl3) δ = 4.14 - 3.96 (m, 1H), 3.66 (s, 6H), 3.08 - 2.99 (m, 2H), 2.94 - 2.88 (m, 2H), 2.33 - 2.26 (m, 7H), 2.14 - 2.01 (m, 4H), 1.98 - 1.89 (m, 2H), 1.83-1.74 (m, 1H), 1.67 - 1.55 (m, 6H), 1.51 - 1.42 (m, 4H), 1.35 - 1.20 (m, 26H), 0.93 - 0.83 (m, 3H). [00590] Step 5: 9-((2,2-difluorononyl)((1-methylpiperidin-4-yl)methyl)amino)heptadecane- 1,17-diol (40e)

[00591] A mixture of dimethyl 9-[2,2-difluorononanoyl-[(1-methyl-4- piperidyl)methyl]amino]heptadecanedioate (40d) (700 mg, 1.11 mmol, 1.0 eq) in THF (5 mL) was degassed and purged with N2 for 3 times, then BH3-Me2S (10 M, 554.77 μL, 5.0 eq) was added at 0 °C via injection and then the mixture was stirred at 50 °C for 10 h under N2. The reaction mixture was quenched by addition HCl (20 mL) at 40 °C and stirred 3 h, The residue was diluted with H2O (10 mL) and extracted with EtOAc (10 mL × 3). The combined organic layers were washed with brine (15 mL × 2), dried over anhydrous sodium sulfate, filtered and concentrated under reduced pressure to give a residue. The residue was purified by flash silica gel chromatography (ISCO®; 12 g SepaFlash® Silica Flash Column, Eluent of 0 ~ 10% MeOH / DCM gradient @ 25 mL / min) to give compound 9-[2,2-difluorononyl-[(1-methyl-4- piperidyl)methyl]amino]heptadecane-1,17-diol (40e) (150 mg, 240.69 μmol, 21.7% yield, 90% purity) as a yellow oil.¹H NMR (400 MHz, CDCl₃) δ = 3.71 - 3.57 (m, 4H), 3.14 - 2.99 (m, 2H), 2.75 - 2.62 (m, 5H), 2.53 -2.32 (m, 4H), 1.91 - 1.68 (m, 6H), 1.66 - 1.38 (m, 12H), 1.37 - 1.23 (m, 28H), 0.97 - 0.79 (m, 3H). [00592] Step 6: 9-((2,2-difluorononyl)((1-methylpiperidin-4-yl)methyl)amino)heptadecane- 1,17-diyl bis(3-butylnonanoate) (40)

40e 40 [00593] To a solution of 9-[2,2-difluorononyl-[(1-methyl-4- piperidyl)methyl]amino]heptadecane-1,17-diol (40e) (120 mg, 213.95 μmol, 1.0 eq), 3- butylnonanoic acid (160.50 mg, 748.82 μmol, 3.5 eq), TEA (108.25 mg, 1.07 mmol, 148.89 μL, 5.0 eq) and DMAP (78.41 mg, 641.84 μmol, 3.0 eq) in DCM (2 mL) was added EDCI (102.54 mg, 534.87 μmol, 2.5 eq) . The mixture was stirred at 25 °C for 10 h. The reaction was concentrated. The residue was purified by prep-HPLC (FA condition; column: CD12-WePure Biotech Phenyl-Hexyl 150 × 25 mm × 7 um; mobile phase: [H₂O (0.225% FA) - THF : CAN = 1 : 3]; gradient: 80% - 100% B over 15.0 min) to give compound [17-(3-butylnonanoyloxy)-9- [2,2-difluorononyl-[(1-methyl-4-piperidyl)methyl]amino]heptadecyl] 3-butylnonanoate (40) (63.45 mg, 66.40 μmol, 31.0% yield, 99.78% purity) as a yellow oil. LCMS: [M+H]⁺: 953.5. ¹H NMR (400 MHz, (CD₃)₂CO-d₆) δ = 4.04 (t, J = 6.4 Hz, 4H), 3.40 - 3.26 (m, 1H), 3.05 - 3.00 (m, 2H), 2.85 - 2.76 (m, 5H), 2.73 - 2.63 (m, 4H), 2.53 - 2.45 (m, 3H), 2.23 (d, J = 6.8 Hz, 4H), 1.88 - 1.77 (m, 4H), 1.64 - 1.58 (m, 4H), 1.53 - 1.44 (m, 6H), 1.39 - 1.24 (m, 62H), 0.95 - 0.84 (m, 15H). LNP Formulation and Characterization: Example A1: General preparation of mRNA lipid formulations [00594] Lipid-encapsulated mRNA particles were prepared by mixing lipids (ionizable cationic lipid: DSPC: cholesterol: PEG-DMG in a 50:10:38.5:1.5 molar ratio) in ethanol (20 mM) with firefly luciferase mRNA (Cynbio, Catalog No. C-2069) dissolved in acetate buffer (pH5, 10 mM) using a microfluidic device (Precision Nanosystems Inc) with a flow ratio of 3:1 (aqueous to organic phase) and a total flow rate of 12 mL/min. The ionizable cationic lipids used in the formulation were selected lipids of Formula I described herein. Ethanol was removed by dialysis against 25 mM tris buffer (pH 7.4, 250 mM sucrose, 63 mM NaCl) overnight at 2-8 °C using a regenerated cellulose membrane (20 kD MWCO). The final formulation was then filtered through a 0.2-μm filter, filled into vials, and stored at −70±5 °C. [00595] The compositions of formulations F1 to F7, made in accordance with this procedure, are listed in Table 2. The N:P ratio of the resulting LNPs was about 6. Example A2: LNP characterization [00596] The frozen formulations from Example A1 were characterized for their mRNA content and percent encapsulation by a RiboGreen assay, and for their particle size and polydispersity index (PDI) by dynamic light scattering on a Wyatt DynaPro Plate Reader. [00597] Particle Size Analysis by Dynamic Light scattering. LNPs were diluted to 10 μg/mL total mRNA in saline, pH 7.4, and transferred into a polystyrene cuvette to measure particle size and polydispersity by dynamic light scattering (Wyatt DynaPro Plate Reader). [00598] Apparent pKa Measurement by TNS Assay. The pKa of each LNP was determined using the TNS binding assay described in J. Heyes, L. Palmer, K. Bremner, I. MacLachlan, J. Controlled Release 2005, 107, 276, the contents of which are incorporated herein by reference. [00599] TNS (6-(p-toluidino)-2-napthalenesulfonic acid) was dissolved in DMSO at a concentration of 0.6 mM. LNPs were diluted with formulation buffer to a total lipid concentration of 0.5 mM. A solution of 20 mM citrate and 150 mM NaCl was made at pH 3.5, 4, 4.5, 5 and 5.5. For pH 6, 6.5, 7, 7.5 and 8, a solution of 20 mM sodium phosphate and 150 mM NaCl was made. A solution of 20 mM Tris HCl and 150 mM NaCl was made for pH 8, 8.5, 9. For pH 9.5, 10, and 10.7, 20 mM carbonate bicarbonate buffer was made. Stock solutions were diluted to reach final well concentrations of 75 μM total lipid and 6 μM TNS at each pH point. Once plated, the plate was covered and incubated for 5 minutes, followed by a 10-minute shake at 37°C in a Synergy H1 plate reader. Fluorescence was measured at an excitation of 321 nm and an emission of 447 nm. Apparent pKa was calculated by taking the relative IC50 value of the pH/RFU data using Graphpad Prism (version 10). [00600] Percent mRNA encapsulation and mRNA concentration. Encapsulation efficiency was measured using a modified Quant-iT RiboGreen assay (Life Technologies). To determine the encapsulation efficiency, samples were diluted in 1X Tris-EDTA (TE, pH 7.5) buffer to a concentration of about 2-10 μg/mL. Additional samples were diluted with 1% Triton X-100 to a concentration of 0.5 μg/mL. RiboGreen dye was diluted 200-fold in 1X TE and added 1:1 (v/v) with sample. Fluorescent signal was quantified using a Synergy H1 plate reader set at 485 nm excitation and 528 nm emission. The standard curve was calculated by linear regression analysis of the fluorescence intensity plotted against the concentration of the standard. RNA encapsulation of LNP samples was determined by comparing the signal of the RNA-binding fluorescent dye RiboGreen in the absence and presence of a detergent (1% Triton X-100). [00601] In the absence of detergent, the signal was generated by accessible (unencapsulated) RNA. In the presence of detergent, the LNP was disrupted, and the measured signal was generated by the total RNA (both encapsulated and nonencapsulated). The encapsulation percentage was calculated using the following equation: Encapsulation efficiency (%) = ((Fluorescence)total − (Fluorescence)unencapsulated)/(Fluorescence)total × 100%. [00602] The results of the analyses of particle size, PDI, and encapsulation efficiency for formulations F1 to F7 are reported in Table 2. Table 2

In Vivo Experiments: Example B1: Administration of mRNA LNPs via intravenous injection [00603] LNP formulations encapsulating Fluc mRNA were injected intravenously via tail vein at 0.5 mg kg⁻¹ to female BALB/c mice (7–8 weeks old). At 6 hours post-dose, animals (with their fur trimmed from the regions of interest for imaging) were dosed with D-Luciferin via intraperitoneal (IP) injection. Mice were anesthetized and imaged using the Caliper IVIS Lumina II System. Tissues were collected for the analysis of luciferase activity post in vivo imaging. The results of the imaging are reported in Table 3. Example B2: Administration of mRNA LNPs via intratracheal instillation (ITr) [00604] LNP formulations encapsulating Fluc mRNA were instilled via intratracheal route at 5 μg (0.1 mg/mL, 50 μL) per mouse to female BALB/c mice (7–8 weeks old). At 6 hours post- dose, animals (with their fur trimmed from the regions of interest for imaging) were dosed with D-Luciferin via intraperitoneal (IP) injection. Mice were anesthetized and imaged using the Caliper IVIS Lumina II System. Tissues were collected for the analysis of luciferase activity post in vivo imaging. The results of the imaging are reported in Table 3. Example B3: Administration of mRNA LNPs via subcutaneous injection (Sc) LNP formulations encapsulating Fluc mRNA were instilled via subcutaneous injection at 6 μg (0.06 mg/mL, 100 μL) per mouse to female BALB/c mice (7–8 weeks old). At 24 hours post- dose, animals (with their fur trimmed from the regions of interest for imaging) were dosed with D-Luciferin via intraperitoneal (IP) injection. Mice were anesthetized and imaged using the Caliper IVIS Lumina II System. Tissues were collected for the analysis of luciferase activity post in vivo imaging. The results of the imaging are reported in Table 3. Table 3

[00605] The teachings of all patents, published applications, and references cited herein are incorporated by reference in their entirety. [00606] While example embodiments have been particularly shown and described, it will be understood by those skilled in the art that various changes in form and details may be made therein without departing from the scope of the embodiments encompassed by the appended claims.

Claims

We claim: 1. A compound of Formula I:

, —O(C═O)—, —(C═O)O—, —C(=O)—, —O—, —S(O)_x—, —S—S—,—NR^aC(═O)—, —C(═O)NR^a—, —NR^aC(═O)NR^a—, —OC(═O)NR^a—, —NR^aC(═O)O—, or a direct bond; each R^a is independently H or C1—C12 aliphatic; each R^b is independently C₁—C₈ aliphatic; each R^c is independently H or C1—C12 aliphatic; each x is independently 0, 1 or 2; M¹ and M² are each independently C₁—C₁₂ aliphatic or a direct bond; G¹ is C₁—C₂ alkylene or a direct bond; G² is —C(═O)S—, —C(═O)O—, —C(═O)—, —C(═O)N(H)—, —C(═O)N(C1—C12 aliphatic)—, —S(=O)2—, —S(=O)(=NH)—, —S(=O)(=N(C1—C12 aliphatic))—, —S(=O)—, —O—, —NH—, —CH₂C(=O)O—, —CH₂CF₂—, —C(=O)CF₂— , or a direct bond; R¹ and R² are each independently H or C1—C16 aliphatic; R³ is C₄—C₂₀ aliphatic optionally substituted with —L³⁰—R³⁰; L³⁰ is —O(C═O)—, —(C═O)O—, —C(=O)—, —O—, —S(O)x3—, —S—S—, —NR^3aC(═O)—, —C(═O)NR^3a—, —NR^3aC(═O)NR^3a—, —OC(═O)NR^3a—, or —NR^3aC(═O)O—; each R^3a is independently H or C₁—C₁₂ aliphatic; R³⁰ is H or C1—C12 aliphatic; x3 is 0, 1, or 2; R⁴ is or , wherein indicates the point of attachment of R⁴ to the nitrogen atom of Formula I; G³ is a direct bond or C1—C12 alkylene; G⁴ is C₂—C₁₂ alkylene; R⁵ is a 4-, 5-, 6-, or 7-membered, carbon-linked, nitrogen-containing heterocycle, wherein each nitrogen atom of the heterocycle is independently substituted with R^d, and the heterocycle is optionally substituted on a carbon atom with R^e; R⁶ and R⁷ are each independently C₁—C₆ aliphatic, or R⁶ and R⁷, taken together with the nitrogen atom to which they are attached, form a 4-, 5-, 6-, or 7-membered, nitrogen- containing heterocycle, wherein each nitrogen atom of the heterocycle other than the nitrogen atom attached to G⁴, R⁶, and R⁷ is independently substituted with R^d, and the heterocycle is optionally substituted on a carbon atom with R^e; each R^d is independently -(C1—C6 aliphatic) or –(C1—C6 aliphatic)OH; each R^e is independently -OH, -(C₁—C₆ aliphatic)OH, or -O(C₁—C₆ aliphatic); and provided: (i) when G² is —S(=O)₂—, either L¹ and L² are both a direct bond or neither L¹ nor L² is a direct bond; and (ii) when G² is a direct bond or —C(═O)—, R⁴ is

.

2. The compound of claim 1, which is a compound of Formula I-a:

, or a pharmaceutically acceptable salt, solvate, or hydrate thereof.

3. The compound of claim 1 or 2, wherein G² is —C(═O)S—, —C(═O)O—, —C(═O)—, —C(═O)N(H)—, —C(═O)N(C₁—C₁₂ aliphatic)—, —S(=O)₂—, —S(=O)—, or a direct bond.

4. The compound of any one of claims 1-3, wherein G² is —C(═O)S—, —C(═O)O—, —C(═O)N(H)—, —C(═O)N(C1—C12 aliphatic)—, —S(=O)2—, or —S(=O)—.

5. The compound of any one of claims 1-4, wherein G² is —C(═O)S— or —S(=O)—.

6. The compound of any one of claims 1-5, wherein R⁵ or the heterocycle formed by R⁶ and R⁷, taken together with the nitrogen atom to which they are attached, is azetidinyl, pyrrolidinyl, piperdinyl, azepanyl, morpholinyl, thiomorpholinyl, or piperazinyl.

7. The compound of any one of claims 1-6, wherein R⁵ is 2-morpholinyl, 3-piperdinyl, 2-pyrrolidinyl or 4-piperdinyl.

8. The compound of any one of claims 1-7, wherein L¹ and L² are each independently

9. The compound of any one of claims 1-8, wherein L¹ and L² are the same.

10. The compound of any one of claims 1-9, which is a compound of formula I-b

or a pharmaceutically acceptable salt, solvate, or hydrate thereof, wherein m and n are each independently an integer from 0 to 5, and the sum of m and n is 2 to 5.

11. The compound of claim 10, wherein the sum of m and n is 4.

12. The compound of claim 10 or 11, wherein m and n are both 2.

13. The compound of any one of claims 1-12, wherein G³ is C1—C5 alkylene.

14. The compound of any one of claims 1-13, wherein R^b is C₁-C₄ alkylene.

15. The compound of any one of claims 1-14, wherein R^c is C1—C12 alkyl.

16. The compound of any one of claims 1-15, wherein G¹ is a direct bond.

17. The compound of any one of claims 1-16, wherein M¹ and M² are the same, and are both C6—C12 aliphatic.

18. The compound of any one of claims 1-17, wherein R¹ and R² are the same, and are C₁—C₁₆ alkyl.

19. The compound of any one of claims 1-18, wherein each R^c, R¹, and R² is the same.

20. The compound of any one of claims 1-19, wherein

21. The compound of any one of claims 1-20, wherein R³ is C₄—C₁₀ alkyl. 22. The compound of any one of claims 1-21, wherein R³ is

,

23. The compound of any one of claims 1, 3-6, 8, 9, and 14-22, wherein R⁴ is . 24. The compound of any one of claims 1, 3-6, 8, 9, and 14-23, wherein G⁴ is C₂—C₅ alkylene; and R⁶ and R⁷ are each C₁—C₃ alkyl and are the same; or R⁶ and R⁷ taken together with the nitrogen atom to which they are attached, form a 4-, 5-, 6-, or 7-membered, nitrogen- containing heterocycle, wherein each nitrogen atom of the heterocycle other than the nitrogen atom attached to G⁴, R⁶, and R⁷ is independently substituted with R^d, and the heterocycle is optionally substituted on a carbon atom with R^e. 25. The compound of any one of claims 1, 3-6, 8, 9, and 14-22, wherein R⁴ is

. 26. A compound of Table 1, or a pharmaceutically acceptable salt, solvate, or hydrate thereof. 27. A lipid nanoparticle comprising the compound of any one of claims 1-26, or a pharmaceutically acceptable salt, solvate, or hydrate thereof, and a therapeutic agent or one or more nucleic acids. 28. A lipid nanoparticle comprising the compound of any one of claims 1-26, or a pharmaceutically acceptable salt, solvate, or hydrate thereof, a steroid, a PEG lipid, and a phospholipid, optionally further comprising a therapeutic agent or one or more nucleic acids. 29. A pharmaceutical composition comprising the compound of any one of claims 1-26, or a pharmaceutically acceptable salt, solvate, or hydrate thereof, or lipid nanoparticle of claim 27 or 28, and a pharmaceutically acceptable carrier. 30. A method of administering a therapeutic agent or nucleic acid to a subject, comprising administering to the subject a lipid nanoparticle of claim 27 or 28.