Novel lipids, lipid compositions containing thereof, and methods of using thereof

Abstract

This disclosure relates to novel lipid compounds containing a head group containing an aromatic or heteroaromatic group and a monoamine, a linker group, and a tertiary amine branched tail group, wherein the chain length counting the total number of atoms forming the chain connecting the head group and the tail group is no more than nine (e.g., is nine). The disclosure also provides a lipid composition comprising these novel lipid compounds and pharmaceutical compositions comprising these lipid compositions. The disclosure additionally provides methods for delivering a therapeutic agent to a cell or a subject and methods of treating a disease or disorder (e.g., a kidney disease or disorder) in a subject employing the lipid composition or pharmaceutical compositions comprising these novel lipid compounds and the therapeutic agent.

Description

NOVEL LIPIDS, LIPID COMPOSITIONS CONTAINING THEREOF, AND METHODS OF USING THEREOF FIELD OF THE INVENTION

[0001] The present disclosure generally relates to novel ionizable lipids, lipid compositions, pharmaceutical compositions, and methods of using thereof for delivery and treatment.BACKGROUND

[0002] Lipid composition such as liposomes and lipid nanoparticles formed from ionizable monoamine-containing lipids can serve as carriers for delivery of therapeutic agents. One of the main challenges is achieving efficient and targeted delivery of the lipid nanoparticles to specific cells or tissues. Delivery of lipid carriers such as lipid nanoparticles face multiple barriers, including clearance by the liver, spleen, and kidneys, which can limit circulation time and reduce therapeutic efficacy, as well as the potential for toxicity and unwanted immune responses arising from their foreign nature and inherent adjuvant properties. There thus continues to be a need in the art for improved lipid carriers employing novel lipid compounds for delivering therapeutic agents, such as nucleic acids, proteins, and small molecule drugs, into targeted cells or tissues, such as kidney, liver, splenic, lymphatic, or marrow cells or tissues.SUMMARY OF THE INVENTION

[0003] One aspect of the invention relates to a compound having the formula of:Ri R / k2R2Ra L 1 o~ X-] L-] -]— RR-]N - LL — NRxr\|_| ' • r\ D l_20— X2— 1_21_ RR2(GI),ORRi Ri ^2 ^2L-i Q — X-j — L — RRM21ML2. B1RH^ ^i\r LL — I NZ>a I-20- ^2 — 1-21 RR2(GII),wherein:RH is arylalkyl, aryl, heteroaryl, or heterocyclyl;each of Ri, Ri', R2, and R2', in each occurrence, is independently H, halo, C1-C3 alkyl, C3-C7 cycloalkyl, or -N(Ra)C(O)Rc; or Ri and Ri', or R2 and R2', together with their adjacent carbon atom, form a ring;nl and n2 are each independently 0, 1, 2, 3, 4, or 5;LEGAL\113833648\6xA is absent, -0-, -S-, -S-S-, -C(Rb)(Rb')-, -C(0)-, -C(0)0-, -0C(0)-, -S(0)->0 0 o oV^N— t V Q— i- 1? 1?Ra,, -P(0)(0Ra)-, -N(Ra)-, -N(Ra)C(0)-, or -C(0)N(Ra)-;0 0xB is -C(0)-, -C(0)0-, -0C(0)-, -S(0)-> or;each of Ra and Ra', in each occurrence, is independently H, C1-C3 alkyl, or C3-C7 cycloalkyl;each of Rb and Rb', in each occurrence, is independently H, OH, C1-C3 alkyl, or C3-C7 cycloalkyl;Rc in each occurrence is independently H, alkyl, or -N(Ra)(Ra');Rq is H, alkyl, or aryl; optionally substituted with one or more of alkyl, halo, or -ORa; LL is C4 alkylene, C4 alkenylene, C4 alkynylene, or-(C(R3)(R3'))mi-Q-(C(R3)(R3'))m2-; or LL, together with Ra and the N atom connected to both B and Ra, forms a linker of the formulaeach of R3 and R3', in each occurrence, is independently H, C1-C3 alkyl, or C2-C3 alkenyl; Q is a divalent C3-C7 cycloalkyl, a divalent bridged cycle, or a divalent heterocycle; ml and m2 are each independently 0, 1, or 2;m3 is 0 or 1;m4 is 1 to 3;Xi and X2are each independently -C(O)-, -C(O)O-, -OC(O)-, -C(O)S-, -SC(O)-, -N(Ra)C(0)-, or -C(0)N(Ra)-;Lio and L20 are each independently a C1-C12 alkylene, optionally substituted by C1-C3 alkyl;Ln and L21 are each independently absent or a Ci-Ce alkylene;c / R-105E— XRR1 is -E-R30 or *Rl1;sI — E -RR2 is -E-R40 orRz1;E is absent, -CH(Ra)-, -CH(Ci-C4alkylene-CO(NH2))-, O, S, arylene, or heteroarylene; R30 and R40 each are independently H, branched or unbranched alkyl, branched or unbranched alkenyl, branched or unbranched alkynyl, bridged cyclyl, aryl, aralkyl, arylalkenyl,- 2 - LEGAL\113833648\6arylalkynyl, aralkoxy, heteroaryl, heteroarylalkyl, heteroarylalkenyl, or heteroarylalkynyl; optionally substituted with one or more of alkyl, halo, or -ORa;Rio and R20 each are independently H or a branched or unbranched, saturated or unsaturated Ci -C12 monovalent hydrocarbon chain; andR11 and R21 each are independently a branched or unbranched, saturated or unsaturated Ci -C12 monovalent hydrocarbon chain.

[0004] Another aspect of the invention relates to a compound having the formula of(PE-I) orwherein:RH is alkyl, arylalkyl, aryl, heteroaryl, or heterocyclyl;each of Ri, Ri', R2, and R2', in each occurrence, is independently H, halo, C1-C3 alkyl, C3- C7 cycloalkyl, or -N(Ra)C(O)Rc; or Ri and Ri', or R2 and R2', together with their adjacent carbon atom, form a ring;nl and n2 are each independently 0, 1, 2, 3, 4, or 5;0 0 xA is absent, -O-, -S-, -S-S-, -C(Rb)(Rb')-, -C(O)-, -C(O)O-, -OC(O)-, -S(O)-,0 0 c> oV V', -P(O)(ORa)-, -N(Ra)-, -N(Ra)C(O)-, or -C(O)N(Ra)-;0 0xB is -C(O)-, -C(O)O-, -OC(O)-, -S(O)-, or;each of Ra and Ra', in each occurrence, is independently H, C1-C3 alkyl, or C3-C7 cycloalkyl;each of Rb and Rb', in each occurrence, is independently H, OH, C1-C3 alkyl, or C3-C7 cycloalkyl;Rc in each occurrence is independently H, alkyl, or -N(Ra)(Ra');Rq is H, alkyl, or aryl; optionally substituted with one or more of alkyl, halo, or -ORa;LEGAL\113833648\6LL is C4 alkylene, C4 alkenylene, C4 alkynylene, or-(C(R3)(R3'))mi-Q-(C(R3)(R3'))m2-; or LL, together with Ra and the N atom connected to both B and Ra, forms a linker of the formulaeach of R3 and R3', in each occurrence, is independently H, C1-C3 alkyl, or C2-C3 alkenyl; Q is a divalent C3-C7 cycloalkyl, a divalent bridged cycle, or a divalent heterocycle; ml and m2 are each independently 0, 1, or 2;m3 is 0 or 1;m4 is 1 to 3;X10 and X20 are each independently -C(O)O-, or -OC(O)-;Xno, X120, X210, and X220 are each independently absent, -C(O)O-, or -OC(O)-, provided that at least one of Xno, Xi2o, X2io, and X220 is not absent;Lio and L20 are each independently a C1-C12 alkylene, optionally substituted by C1-C3 alkyl;Ln, L21, Lno, L120, L210, and L220 are each independently absent or a Ci-Ce alkylene; and Rm, R121, R211, and R221 each are independently a branched or unbranched, saturated or unsaturated Ci -C12 monovalent hydrocarbon chain;with the proviso that, when RH is alkyl, A is not absent and not CH2.

[0005] Also provided herein is a lipid composition comprising a lipid component and the compounds as described herein in the above aspects relating to the lipid compounds. Suitable compounds are all the lipid compounds described herein, including all the compounds of formulas (GI), (GII), (GL), (GIL), (PE -I), and / or (PE-II) and any of the compounds belonging to any subgenus formulas or species under formula (GI), (GII), (GL), (GIL), (PE-I), or (PE-II), described herein. The lipid composition may be in the form of a liposome or lipid nanoparticle. The lipid composition (e.g., liposome or LNP) described herein can be used for the delivery of a therapeutic agent to a living organism, such as to a human subject. The therapeutic agent may be loaded to the lipid composition, e.g., encapsulated within the lipid composition (e.g., liposome or LNP). In some embodiments, the lipid composition further comprises a therapeutic agent.

[0006] Another aspect of the invention relates to a pharmaceutical composition comprising the lipid composition (e.g., liposome or LNP), which contains the lipid compound described herein (including one or more compounds of formula (GI), (GII), (GT), (GIT), (PE-I), or (PE-II); or any of the compounds belonging to any subgenus formulas or species under formula (GI), (GII), (GT), (GIT), (PE-I), or (PE-II) disclosed herein) and which is loaded with the therapeutic agent described herein, and a pharmaceutically acceptable carrier.

[0007] Also provided herein are methods for delivering a therapeutic agent to a cell or a subject- 4 - LEGAL\113833648\6(e.g., a patient) in need thereof, comprising contacting the cell with, or administering to the subject, the lipid composition according to any of the presently disclosed embodiments, or the pharmaceutical composition according to any of the presently disclosed embodiments, wherein the lipid composition comprises a therapeutic agent.

[0008] Another aspect of the invention relates to a method of treating a disease or disorder in a subject in need of such treatment, the method comprising: administering to the subject lipid composition according to any of the presently disclosed embodiments, or the pharmaceutical composition according to any of the presently disclosed embodiments, wherein the lipid composition comprises a therapeutic agent.

[0009] Also provided herein is a method for treating a kidney disease or kidney disorder, the method comprising: administering to the subject the lipid composition according to any of the presently disclosed embodiments, or the pharmaceutical composition according to any of the presently disclosed embodiments, wherein the lipid composition comprises a therapeutic agent.BRIEF DESCRIPTION OF THE DRAWINGS

[0010] FIGs. 1A-1C illustrate the flow cytometry results for a high-throughput, barcoding based, screen in mouse using the LNP composition containing exemplary ionizable lipids described herein, with the LNP encapsulating aVHH mRNA at 2.0 mg / kg, in the liver (FIG.1 A), spleen (FIG. IB), and bone marrow (FIG. 1C). FIGs. 1D-1E depict the diameter (nm) (FIG. ID) and polydispersity index (PDI) (FIG. IE) of the LNP composition used in the screen containing exemplary ionizable lipids described herein, with the LNP encapsulating aVHH mRNA at 2.0 mg / kg.

[0011] FIGs. 2A-2C show the flow cytometry results for individual LNP formulations selected from the screen discussed in FIGs. 1 A-1C in mouse using the LNP formulations containing exemplary ionizable lipids described herein, with the LNP encapsulating aVHH mRNA at 0.2 mg / kg, in the liver (FIG. 2A), spleen (FIG. 2B), and bone marrow (FIG. 2C).

[0012] FIG. 3 depict the parameters of the individual LNP formulations discussed in FIG. 2, selected from the screen discussed in FIGs. 1A-1C, showing the diameter (nm) and the encapsulation efficiency (%) for respective LNPs tested.

[0013] FIGs. 4A-4B depict the parameters of the LNP composition containing exemplary ionizable lipids described herein, with the LNP encapsulating aVHH mRNA at 0.25 mg / kg. FIG.4A shows the diameter (nm) and polydispersity index (PDI) for respective LNPs tested. FIG. 4B shows the encapsulation efficiency (%) for respective LNPs tested.

[0014] FIGs. 5 A-5F show the flow cytometry results indicating the in vivo potency of the LNP composition containing exemplary ionizable lipids encapsulating aVHH mRNA at 0.25 mg / kg in- 5 - LEGAL\113833648\6various cells in mice, after a single intravenous administration of the LNP composition to mice. FIG. 5A shows % aVHH+ in splenic CD8 T cells. FIG. 5B shows the aVHH mean fluorescence intensity (MFI) in splenic CD8 T cells. FIG. 5C shows % aVHH+ in bone marrow LSKs. FIG.5D shows aVHH MFI in bone marrow LSKs. FIG. 5E shows % aVHH+ in hepatic CD31 ECs. FIG. 5F shows aVHH MFI in hepatic CD31.

[0015] FIGS. 6A-6B depict the parameters of the LNP composition containing exemplary ionizable lipids described herein, with the LNP encapsulating aVHH mRNA at 0.1 mg / kg. FIG.6A shows the diameter (nm) and PDI for respective LNPs tested. FIG. 6B shows the encapsulation efficiency (%) for respective LNPs tested.

[0016] FIGs. 7A-7D show the flow cytometry results indicating the in vivo potency of the LNP composition containing exemplary ionizable lipids encapsulating aVHH mRNA at 0.1 mg / kg in various cells in mice, after a single intravenous administration of the LNP composition to mice. FIG. 7A shows % aVHH+ in splenic CD8 T cells. FIG. 7B shows the aVHH mean fluorescence intensity (MFI) in splenic CD8 T cells. FIG. 7C shows % aVHH+ in bone marrow LSKs. FIG.7D shows aVHH MFI in bone marrow LSKs.

[0017] FIGs. 8A-8B depict the parameters of the CD5-targeted LNP composition containing exemplary ionizable lipids described herein, with the CD5-targeted LNPs encapsulating mCherry mRNA at 0.5 mg / kg. FIG. 8A shows the diameter (nm) and PDI for respective CD5-targeted LNPs tested. FIG. 8B shows the encapsulation efficiency (%) for respective CD5-targeted LNPs tested.

[0018] FIGs. 9A-9H show the flow cytometry results indicating the in vivo potency of the CD5-targeted LNP composition containing exemplary ionizable lipids encapsulating mCherry mRNA at 0.5 mg / kg in various cells in mice, after a single intravenous administration of the CD5-targeted LNP composition to mice. FIG. 9A shows % mCherry+ in splenic CD8 T cells. FIG.9B shows the mCherry mean fluorescence intensity (MFI) in splenic CD8 T cells. FIG. 9C shows % mCherry+ in splenic CD19 B cells. FIG. 9D shows mCherry MFI in CD19 B cells. FIG. 9E shows % mCherry+ in bone marrow CD8 T cells. FIG. 9F shows mCherry MFI in bone marrow CD8 T cells. FIG. 9G shows % mCherry+ in PBMC CD8 T cells. FIG. 9H shows mCherry MFI in PBMC CD8 T cells.

[0019] FIGs. 10A-10B depict the parameters of the CD5-targeted LNP composition containing exemplary ionizable lipids described herein, with the CD5-targeted LNP encapsulating mCherry mRNA at 0.5 mg / kg. FIG. 10A shows the diameter (nm) and PDI for respective CD5-targeted LNPs tested. FIG. 10B shows the encapsulation efficiency (%) for respective CD5-targeted LNPs tested.

[0020] FIGs. 11 A-l IF show the flow cytometry results indicating the in vivo potency of the- 6 - LEGAL\113833648\6CD5-targeted LNP composition containing exemplary ionizable lipids encapsulating mCherry mRNA at 0.5 mg / kg in various cells in mice, after a single intravenous administration of the CD5-targeted LNP composition to mice. FIG. 11 A shows % mCherry+ in splenic CD8 T cells. FIG. 1 IB shows the mCherry mean fluorescence intensity (MFI) in splenic CD8 T cells. FIG.11C shows % mCherry+ in bone marrow CD8 T cells. FIG. 1 ID shows mCherry MFI in bone marrow CD8 T cells. FIG. 1 IE shows % mCherry+ in PBMC CD8 T cells. FIG. 1 IF shows mCherry MFI in PBMC CD8 T cells.

[0021] FIGs. 12A-12B depict the parameters of the LNP composition containing exemplary ionizable lipids described herein, with the LNP encapsulating hEPO mRNA at 0.2 mg / kg. FIG.12A shows the diameter (nm) and PDI for respective LNPs tested. FIG. 12B shows the encapsulation efficiency (%) for respective LNPs tested.

[0022] FIG. 13 shows the hEPO quantification results indicating the in vivo potency of the LNP composition containing exemplary ionizable lipids encapsulating hEPO mRNA at 0.2 mg / kg in serum of mice, after a single intravenous administration of the LNP composition to mice.

[0023] FIGs. 14A-14B depict the parameters of the LNP composition containing exemplary ionizable lipids described herein, with the LNP encapsulating aVHH mRNA at 1 mg / kg. FIG.14A shows the diameter (nm) and PDI for respective LNPs tested. FIG. 14B shows the encapsulation efficiency (%) for respective LNPs tested.

[0024] FIG. 15 shows the in vivo potency of the LNP composition containing exemplary ionizable lipids encapsulating aVHH mRNA at 1 mg / kg in kidney of mice, after a single intravenous administration of the LNP composition to mice. The in vivo potency of the LNP composition containing exemplary ionizable lipids encapsulating aVHH mRNA at 1 mg / kg in kidney was assessed by quantifying immunofluorescence staining (IF) of the LNP composition in kidney renal cortex against aVHH-HA protein using a labeled HA-tag.

[0025] FIGs. 16A-16B depict the parameters of the LNP composition containing exemplary ionizable lipids described herein, with the LNP encapsulating Cas9 mRNA and sgB2M at 2 x 2 mg / kg. FIG. 16A shows the diameter (nm) and PDI for respective LNPs tested at different dose levels. FIG. 16B shows the encapsulation efficiency (%) for respective LNPs tested at different dose levels.

[0026] FIGs. 17A-17B show the in vivo potency of the LNP composition containing exemplary ionizable lipids encapsulating Cas9 mRNA and B2M sgRNA in kidney and liver of mice, after two intravenous administrations of the LNP composition at each dose of 2 mg / kg Cas9 mRNA and B2M sgRNA, with two doses three days apart. FIG. 17A shows the study timeline and dosing regimen of the LNP composition. FIG. 17B shows the % editing of whole kidney and whole liver tissues after background subtraction of editing in PBS mice.- 7 - LEGAL\113833648\6

[0027] FIG. 18 shows the in vivo biodistribution of the LNP compositions containing exemplary ionizable lipids carrying fLuc mRNA at 0.5 mg / kg in various organs of mice, after a single intravenous administration of the LNP composition to mice. The LNP compositions compared include untargeted LNPs as well as CD5-targeted LNPs, as labeled.DETAILED DESCRIPTION OF THE INVENTION

[0028] Disclosed herein are novel ionizable lipids containing a head group containing an aromatic or heteroaromatic group and a monoamine, a linker group, and a tertiary amine branched tail group, wherein the chain length counting the total number of atoms forming the chain connecting the head group and the tail group is no more than nine (e.g., is nine). These novel ionizable lipid compounds are useful for formation of lipid composition (such as a lipid nanoparticle (LNP) or a liposome). These lipid compositions employing the novel ionizable lipids may have properties advantageous for delivering a therapeutic agent (such as a nucleic acid molecule) to cells. For instance, the lipid compositions employing the novel ionizable lipid compounds may preferentially deliver a therapeutic agent (such as a nucleic acid, e.g., mRNA) to immune cells or stem cells in various tissues in vivo with or without targeting ligands. The examples show that exemplary lipid compositions employing exemplary ionizable lipid compounds may preferentially deliver a therapeutic agent (such as a nucleic acid, e.g., mRNA) to liver or non-liver cells (kidney, splenic, lymphatic, or marrow cells) at clinically relevant doses in vivo with or without targeting ligands.

[0029] The disclosure thus provides the lipid compositions comprising these novel lipids. The disclosure also provides pharmaceutical compositions comprising these lipid compositions. Additionally, the disclosure provides methods for delivering a therapeutic agent to a cell or a subject by contacting the call or administering to the subject the lipid compositions or the pharmaceutical compositions comprising these lipid compositions, which contain the therapeutic agent.Novel Ionizable Lipids

[0030] One aspect of the invention relates to a compound having the formula of:Ri RiK2R2Ra L10-X1-L11-RR1R^ | / N-LL-<HL20X2L21_ RR2(GI),ORLEGAL\113833648\6Ri ^1 ^2 ^2Acj11BLw~xi—Ln-RR1RH'' ^LL — NZL20-X2— L21- RR(GII),wherein:RH is arylalkyl, aryl, heteroaryl, or heterocyclyl;each of Ri, Ri', R2, and R2', in each occurrence, is independently H, halo, C1-C3 alkyl, C3-C7 cycloalkyl, or -N(Ra)C(O)Rc; or Ri and Ri', or R2 and R2', together with their adjacent carbon atom, form a ring;nl and n2 are each independently 0, 1, 2, 3, 4, or 5;0 0 x A is absent, -O-, -S-, -S-S-, -C(Rb)(Rb')-, -C(O)-, -C(O)O-, -OC(O)-, -S(O)-, * o o o oV■A, -P(O)(ORa)-, -N(Ra)-, -N(Ra)C(O)-, or -C(O)N(Ra)-;0 0xB is -C(O)-, -C(O)O-, -OC(O)-, -S(O)-, or;each of Ra and Ra', in each occurrence, is independently H, C1-C3 alkyl, or C3-C7 cycloalkyl;each of Rb and Rb', in each occurrence, is independently H, OH, C1-C3 alkyl, or C3-C7 cycloalkyl;Rc in each occurrence is independently H, alkyl, or -N(Ra)(Ra');Rq is H, alkyl, or aryl; optionally substituted with one or more of alkyl, halo, or -ORa; LL is C4 alkylene, C4 alkenylene, C4 alkynylene, or-(C(R3)(R3'))mi-Q-(C(R3)(R3'))m2-; or LL, together with Ra and the N atom connected to both B and Ra, forms a linker of the formulaeach of R3 and R3', in each occurrence, is independently H, C1-C3 alkyl, or C2-C3 alkenyl; Q is a divalent C3-C7 cycloalkyl, a divalent bridged cycle, or a divalent heterocycle; ml and m2 are each independently 0, 1, or 2;m3 is 0 or 1;m4 is 1 to 3;Xi and X2are each independently -C(O)-, -C(O)O-, -OC(O)-, -C(O)S-, -SC(O)-, -N(Ra)C(O)- or -C(O)N(Ra)-;- 9 - LEGAL\113833648\6Lio and L20 are each independently a C1-C12 alkylene, optionally substituted by C1-C3 alkyl;Ln and L21 are each independently absent or a Ci-Ce alkylene;R10RR1 is -E-R30 or R11.R2ORR2 is -E-R40 or R2I.E is absent, -CH(Ra)-, -CH(Ci-C4alkylene-CO(NH2))-, O, S, arylene, or heteroarylene; R30 and R40 each are independently H, branched or unbranched alkyl, branched or unbranched alkenyl, branched or unbranched alkynyl, bridged cyclyl, aryl, aralkyl, arylalkenyl, arylalkynyl, aralkoxy, heteroaryl, heteroarylalkyl, heteroarylalkenyl, or heteroarylalkynyl; optionally substituted with one or more of alkyl, halo, or -ORa;Rio and R20 each are independently H or a branched or unbranched, saturated or unsaturated Ci -C12 monovalent hydrocarbon chain; andR11 and R21 each are independently a branched or unbranched, saturated or unsaturated Ci -C12 monovalent hydrocarbon chain.^Rw

[0031] In formula (GI) or (GII), RR1 is -E-R30 orRl1, and RR2 is -E-R40 or R10R20 _ £.Rz1. In some embodiments, RR1 is ’11. In some embodiments, RR1 is S ^R20-E-R30. In some embodiments, RR2 is21. In some embodiments, RR1 is -E-R40.

[0032] In some embodiments, RR1 is -E-R30, and RR2 is -E-R40. In some embodiments, RR1 R20 R10 is -E-R30, and RR2 is R21 In some embodiments, RR1 isRl1, and RR2 is,R20 <— E. —E— R40. R2I In some embodiments, RR1 IS ^11, and RR2 is

[0033] E is absent, -CH(Ra)-, -CH(CI-C4 alkylene-CO(NH2))-, O, S, arylene, or heteroarylene, wherein Ra is H, C1-C3 alkyl, or C3-C7 cycloalkyl. In some embodiments, E is absent, -CH(Ra)-, - CH(Ci-C4alkylene-CO(NH2))-, O, S, arylene, or heteroarylene. In some embodiments, Ra is H or C1-C3 alkyl (e.g., methyl). E for RR1 and E for RR2 may be the same or different.

[0034] R30 and R40 each are independently H, branched or unbranched alkyl, branched or unbranched alkenyl, branched or unbranched alkynyl, bridged cyclyl, aryl, aralkyl, arylalkenyl, arylalkynyl, aralkoxy, heteroaryl, heteroarylalkyl, heteroarylalkenyl, or heteroarylalkynyl; optionally substituted with one or more of alkyl, halo, or -ORa, In some embodiments, R30 and - 10 - LEGAL\113833648\6Rio each are independently H, branched or unbranched alkyl, branched or unbranched alkenyl, branched or unbranched alkynyl, bridged cyclyl, phenyl, phenylalkyl, phenylalkynyl, phenyloxy, heteroaryl; optionally substituted with one or more of C1-C3 alkyl. R30 and R40 may be the same or different.

[0035] In some embodiments, the compound has the formula ofp p R p Ra^*Rio\ / 1\ / 2I / L10-X1— Ln—AX n1AX n2 Jj —LL—NZ R_HR / \Y| / AoL20-A2 — l-2-l -R21(GT), orRj ^1 2 ^2 RAZn 1 BLio-X1 —Ln —AA'J'>" N'Z\[_L - ITRnI \ ^^201XL20-X2-L21— <5a21(GIF), wherein the definitions for the variables are the same as those described herein for formulas (GI) and / or (GII).

[0036] Another aspect of the invention relates to a compound having the formula ofwherein:RH is alkyl, arylalkyl, aryl, heteroaryl, or heterocyclyl;each of Ri, Ri', R2, and R2', in each occurrence, is independently H, halo, C1-C3 alkyl, C3-C7 cycloalkyl, or -N(Ra)C(O)Rc; or Ri and Ri', or R2 and R2', together with their adjacent carbon atom, form a ring;nl and n2 are each independently 0, 1, 2, 3, 4, or 5;0 0 xA is absent, -O-, -S-, -S-S-, -C(Rb)(Rb')-, -C(O)-, -C(O)O-, -OC(O)-, -S(O)-,0 0 0 0VY "" N— l ^Q—1 s 1 §, -P(O)(ORa)-, -N(Ra)-, -N(Ra)C(O)-, or -C(O)N(Ra)-;0 0xB is -C(O)-, -C(O)O-, -OC(O)-, -S(O)-, or *;- 11 - LEGAL\113833648\6each of Ra and Ra', in each occurrence, is independently H, C1-C3 alkyl, or C3-C7 cycloalkyl;each of Rb and Rb', in each occurrence, is independently H, OH, C1-C3 alkyl, or C3-C7 cycloalkyl;Rc in each occurrence is independently H, alkyl, or -N(Ra)(Ra');Rq is H, alkyl, or aryl; optionally substituted with one or more of alkyl, halo, or -ORa; LL is C4 alkylene, C4 alkenylene, C4 alkynylene, or-(C(R3)(R3'))mi-Q-(C(R3)(R3'))m2-; or LL, together with Ra and the N atom connected to both B and Ra, forms a linker of the formulav'm3each of R3 and R3', in each occurrence, is independently H, C1-C3 alkyl, or C2-C3 alkenyl;Q is a divalent C3-C7 cycloalkyl, a divalent bridged cycle, or a divalent heterocycle; ml and m2 are each independently 0, 1, or 2;m3 is 0 or 1;m4 is 1 to 3;X10 and X20 are each independently -C(O)O-, or -OC(O)-;Xno, X120, X210, and X220 are each independently absent, -C(O)O-, or -OC(O)-, provided that at least one of Xno, Xi2o, X2io, and X220 is not absent;Lio and L20 are each independently a C1-C12 alkylene, optionally substituted by C1-C3 alkyl;Ln, L21, Lno, L120, L210, and L220 are each independently absent or a Ci-Ce alkylene; and Rm, R121, R211, and R221 each are independently a branched or unbranched, saturated or unsaturated Ci -C12 monovalent hydrocarbon chain;with the proviso that, when RH is alkyl, A is not absent and not CH2.

[0037] In some embodiments, for the compounds of formula (GI), (GII), (GL), (GIL), (PE -I), or (PE-II), or the compounds of the subgenus formulas under formula (GI), (GII), (GL), (GIL), (PE-L1o — — \ I), or (PE-II), the total number of atoms forming the chain connecting RH- toL20—is no more than nine. That is to say, the chain length counting the number of the atoms on the chain between RH and N, not including RH and N, is no more than 9. In some embodiments, for the compounds of formula (GI), (GII), (GI'), (GII'), (PE-I), or (PE-II), or the compounds of the subgenus formulas under formula (GI), (GII), (GI'), (GII'), (PE-I), or (PE-II), the total number of- 12 - LEGAL\113833648\6,Lio —- NZatoms forming the chain connecting RH- toL20—is nine.

[0038] For (GI), the chain length would consider all the atoms constituting the chain of, i.e., nl+NA+n2+ NB +1 (i. e., N atom) +NLL (wherein NA, NB, and NLL each are the number of atoms for A, B, and LL, respectively, constituting the chain). For iHN; FTinstance, for a compound having a structure of, the chain length would H•^S^. Nconsider all the atoms constituting the chain of °, wherein nl is 0, NA is 1 (i.e., A is S atom), n2 is 2 (i.e., 2 C atoms), NB is 1 (i.e., B is -C(O)-, but only C atom constitutes the chain), NLL is 4 (i.e., LL is C4 alkylene, 4 C atoms). Thus, the chain length is nl+NA+n2+ NB +1 +NLL = 9.

[0039] In some embodiments, in formula (GI), (GII), (GI'), (GIF), (PE -I), or (PE -II), or the subgenus formulas under formula (GI), (GII), (GI'), (GII'), (PE -I), or (PE -II), LL is C4 alkylene, C4 alkenylene, C4 alkynylene, or -(C(R3)(R3'))mi-Q-(C(R3)(R3'))m2-. In some embodiments, LL, together with Ra and the N atom connected to both B and Ra, forms a linker of the formula

[0040] In some embodiments, LL is a C4 alkylene, C4 alkenylene, C4 alkynylene, or -(CH2)mi- Q-CH2)m2-, wherein:Q is a divalent C4-C6 cycloalkyl or a divalent bridged cycle; andml and m2 are each independently 0 or 1.4^^

[0041] In some embodiments, LL is a C4 alkylene. In some embodiments, LL is.

[0042] In some embodiments, LL is a C4 alkenylene. In some embodiments, LL is

[0043] In some embodiments, LL is a C4 alkynylene. In some embodiments, LL is

[0044] In some embodiments, LL is -(CH2)mi-Q-CH2)m2- In some embodiments, LL is- 13 - LEGAL\113833648\6

[0045] In some embodiments,

[0046] In some embodiments, for the compounds falling under formula (GI) or (GII), the compound may have the formula of:(GII-LII).

[0047] In these formulas, Q’ is -CH2-CH2-, -CH=CH-, -C=C-, a divalent C3-C7 cycloalkyl or a divalent bridged cycle; ml and m2 are each independently 0 or 1; m3 is 0 or 1; and / or m4 is 1 to 3. The definitions for the other variables are the same as those described herein for formulas (GI), (GII), (GP), and / or (GIF).

[0048] In some embodiments, for the compounds falling under formula (GI) or (GII), compound may have the formula of:(GII-LIIa). The definitions for the variables are the same as those described herein for formulas (GI), (GII), (GP), and / or (GIF).

[0049] In some embodiments, for the compounds falling under formula (PE-I) or (PE-II), the compound may have the formula of:^LII0-X11Q-R111R1 Ri'R2 R2' Ra L10-X10-L11— <... / L120-A120-K121Q'r\|_ | ' • r\ D ml V)m2 \20_X20_L21^L210“X210“R211L220-X220-R221 (PE-LI-1)- 14 - LEGAL\113833648\6' R ^Lno-Xiio-RiiiL10-X10—L11 — <^|Lv D120~A120-r<121.v. ^L21Q—X210“R211L20-A20— L21 -L22O-X22O- R221 (PE-LI-2), orRl R-!R2 ^2 I Y I ^^-110-^110-^111 / XT1jf)4n2BQ.N / 10 10 11 L120-X120-Rl21Rh A n / X L ^^_ / L20-A20-L21 —L210-X210-R211aL220-X220-R221 (PE-LII). In these formulas, Q’ is -CH2-CH2-, -CH=CH-, -C=C-, a divalent C3-C7 cycloalkyl or a divalent bridged cycle; ml and m2 are each independently 0 or 1; m3 is 0 or 1; and / or m4 is 1 to 3. The definitions for the other variables are the same as those described herein for formulas (PE-I) and / or (PE-II).

[0050] In some embodiments, for the compounds falling under formula (PE-I) or (PE-II), compound may have the formula of:R1 R1'r2R2' Ra ^. L110-X110-R11 1M1Mn2HO-xw-L11 —vPR^ Nr ^B-L120— A120'-f'121N..v. ^^-210-^210-^211L20~A20~L21 —L220-X220-R221 (PE-LIa), orL110-X11Q-R111L-IO- xio~ L-11 —L120— AY120-DK121.. Y I ^'-21O-X21O-R211Ra'L20-A20-L21 -L220-X220-R221 (PE-LIIa).

[0051] In some embodiments, for the compounds of formula (GI), (GII), (GT), (GIT), (PE-I), or (PE-II), or the compounds of the subgenus formulas under formula (GI), (GII), (GT), (GIT), (PE-I), or (PE-II), each of Ri, Ri', R2, and R2', in each occurrence, is independently H, halo, C1-C3 alkyl, C3-C7 cycloalkyl, or -N(Ra)C(O)Rc. Rc in each occurrence is independently H, alkyl, or -N(Ra)(Ra'). In some embodiments, Rc in each occurrence is independently H or CH3. In some embodiments, each of Ri, Ri', R2, and R2', in each occurrence, is independently H, halo, CH3, -NHC(O)CH3. In one embodiment, each of Ri, Ri', R2, and R2' is H.

[0052] In some embodiments, Ri and Ri', or R2 and R2', together with their adjacent carbon atom, form a ring. In some embodiments, R2 and R2', together with their adjacent carbon atom, form a ring.

[0053] In some embodiments, for the compounds of formula (GI), (GII), (GI'), (GII'), (PE-I), or (PE-II), or the compounds of the subgenus formulas under formula (GI), (GII), (GI'), (GII'), (PE-I), or (PE-II), nl and n2 are each independently 0, 1, 2, 3, 4, or 5. For instance, nl and n2 are each independently 0, 1, 2, or 3. In some embodiments, nl is 0 or 1. In some embodiments, n2 is- 15 - LEGAL\113833648\60, 1, 2, or 3.

[0054] In some embodiments, for the compounds of formula (GI), (GII), (GI'), (GIF), (PE -I), or (PE -II), or the compounds of the subgenus formulas under formula (GI), (GII), (GI'), (GIF), (PE- 0 0 x I), or (PE -II), A is absent, -O-, -S-, -S-S-, -C(Rb)(Rb')-, -C(O)-, -C(O)O-, -OC(O)-, -S(O)-, o o o oV■A, -P(O)(ORa)-, -N(Ra)-, -N(Ra)C(O)-, or -C(O)N(Ra)-. In someembodiments, A is absent, -O-, -S-, -S-S-, -CH2-, -C(O)-, -C(O)O-, -OC(O)-, -S(O)-, o o hr X M, -P(O)(ORa)-, -NH-, -N(CH3)-, -NHC(O)-, -CH(OH)-, or -C(O)N(Ra)-. Each of Ra and Ra', in each occurrence, is independently H, C1-C3 alkyl, or C3-C7 cycloalkyl. Each of Rb and Rb', in each occurrence, is independently H, OH, C1-C3 alkyl, or C3-C7 cycloalkyl. Rq is H, alkyl, or aryl; optionally substituted with one or more of alkyl, halo, or -ORa. In some embodiments, Ra is H, C1-C3 alkyl (e.g., methyl). In some embodiments, Rqis Ci-C3 alkyl or phenyl optionally substituted with one or more alkyl or halo.

[0055] In some embodiments, for the compounds of formula (GI), (GII), (GE), (GIF), (PE -I), or (PE -II), or the compounds of the subgenus formulas under formula (GI), (GII), (GE), (GIF), (PE- 0 0xI), or (PE -II), B is -C(O)-, -C(O)O-, -OC(O)-, -S(O)-, or In some embodiments, B is O OC(O). In some embodiments,B is.

[0056] In some embodiments, in the Head group, each of Ri, Ri', R2, and R2', in each occurrence, is independently H, halo, CH3, -NHC(O) CH,; or R2and R2', together with their adjacent carbon atom, form a ring; nl and n2 are each independently 0, 1, 2, or 3; A is absent, O O C) O 0 0 vr V. Xa -0-, -S-, -S-S-, -CH2-, -C(0)-, -C(0)0-, -0C(0)-, -S(0)->, -P(O)(ORa)-, -NH-, -N(CH3)-, -NHC(O)-, -CH(OH)-, or -C(O)N(Ra)-; wherein Ra is H, C1-C3 alkyl, and Rqis C1-C3 alkyl or phenyl optionally substituted with one or more alkyl O Oor halo; and / or B is -C(O)-, -OC(O)-, or- 16 - LEGAL\113833648\6

[0057] In some embodiments, in the Head group, each of Ri, Ri', R2, and R2' is H; nl is 0 or 1; and n2 is 0, 1, 2, or 3.

[0058] In some embodiments, for the compounds of formula (GI), (GII), (GT), or (GIT), or the compounds of the subgenus formulas under formula (GI), (GII), (GP), or (GIP), RH is arylalkyl, aryl, heteroaryl, or heterocyclyl.

[0059] In some embodiments, for the compounds of formula (PE -I) or (PE-II), or the compounds of the subgenus formulas under formula (PE -I) or (PE-II), RH is alkyl, arylalkyl, aryl, heteroaryl, or heterocyclyl. In some embodiments, RH is alkyl.

[0060] In some embodiments, RH is arylalkyl, phenyl, biphenyl, pyrimidinyl or dioxohexahydropyrimidinyl, pyridyl, pyridinyl, pyrrolyl, imidazolyl, pyrazolyl, triazolyl, tetrazolyl, triazolyl, thiophenyl, indolyl, dioxoisoindolinyl, dihydrobenzodioxinyl, dihydrobenzodioxepanyl, dihydrobenzofuranyl, thienopyrrolyl, benzimidazolyl, dihydroindenyl, naphthalenyl, or oxathianyl or sulfoxidized oxathianyl. These above listed groups for RH may be optionally substituted with one or more of alkyl, halo, haloalkyl, phenyl, heteroaryl, -ORa, -C(Rb)(Rb')ORa, -C(O)N(Ra)(Ra'), -SCERc, -NO2, -B(ORa)2, -ON, wherein two or more substituents, when present, may form a fused ring with the main ring of RH. RC in each occurrence is independently H, alkyl, or -N(Ra)(Ra'). In some embodiments, Rc in each occurrence is independently H or CH3. Rd is H, alkyl, or phenyl.HR. R4 is H or C1-C3 alkyl. Each of R5 and R5', in each occurrence, is independently aryl or heteroaryl, optionally substituted with one or more of alkyl, halo, haloalkyl, or -ORe. Re is, in each occurrence, independently H, alkyl, halo, haloalkyl, phenyl, heteroaryl, -OR7, -(C(R4)(R4'))SOR7, -C(O)N(R4)(R4'), -NO2, -ON, -SO2R4, -B(OR7)2, or =0. R7is H, C1-C3 alkyl,or phenyl.is a 5-, 6-, or 7- member aromatic or non-aromatic ring fused to the phenyl ring,HRoptionally containing 1 to 2 of O, S, or N on the ring.is a 5- or 6- member heteroaromatic or heterocycle containing 1 to 4 of S or N on the ring, s and t are each independently 1 to 3.(Re)t

[0062] In some embodiments, RH is. In some embodiments, Re is H, alkyl, halo, haloalkyl, -OH, -OCH3, -C(O)NH2, -NO2, -ON, -SO2CH3 or -B(OH)2.- 17 - LEGAL\113833648\6LEGAL\113833648\6H '- 19 - LEGAL\113833648\6N< H 'Cl 9- 20 - LEGAL\113833648\6LEGAL\113833648\6&wherein I represents the connection to LL.

[0065] In some embodiments, for the compounds of formula (GI), (GII), (GP), or (GIF), or the compounds of the subgenus formulas under formula (GI), (GII), (GP), or (GIP), Xi and X2 are each independently -C(O)-, -C(O)O-, -OC(O)-, -C(O)S-, -SC(O)-, -N(Ra)C(O)-, or -C(O)N(Ra)-, wherein Ra is H, C1-C3 alkyl, or C3-C7 cycloalkyl. In some embodiments, Ra is H or C1-C3 alkyl (e.g., methyl). In some embodiments, Xi and X2 are each independently -C(O)O-, -OC(O)-, -NHC(O)-, or -C(O)NH-. Xi and X2 may be the same or different.

[0066] In some embodiments, for the compounds of formula (GI), (GII), (GP), or (GII'), or the compounds of the subgenus formulas under formula (GI), (GII), (GP), or (GIP), Lio and L20 are each independently a C1-C12 alkylene, optionally substituted by C1-C3 alkyl. In some embodiments, Lio and L20 are each independently a C4-C8 alkylene, optionally substituted by methyl. In some embodiments, Lio and L20 are each independently a linear C5-C7 alkylene. In some embodiments, Lio and L20 are each independently a linear C5 or C7 alkylene. Lio and L20 may be the same or different.

[0067] In some embodiments, for the compounds of formula (GI), (GII), (GP), or (GII'), or the compounds of the subgenus formulas under formula (GI), (GII), (GP), or (GIP), Ln and L21 are each independently absent or a Ci-Ce alkylene. In some embodiments, Ln and L21 are each independently absent or a C1-C3 alkylene. In some embodiments, Ln and L21 are each independently absent or a C2 alkylene. Ln and L21 may be the same or different.

[0068] For the compounds of formula (GI) or (GII), the variables RR1, RR2, and E, and various embodiments relating to these variables have been described herein. Those embodiments when RR1 is -E-R30 and when RR2 is -E-R40 have been described herein. In some embodiments, for the compounds of formula (GI), (GII), (GP), or (GIP), or the compounds of the subgenus- 22 - LEGAL\113833648\6< / R-ioi-E^<formulas under formula (GI), (GII), (GT), or (GIT), where RR1 isRl1, or where RR2 isRz1, Rio and R20 each are independently H or a branched or unbranched, saturated or unsaturated Ci -C12 monovalent hydrocarbon chain, and Rn and R21 each are independently a branched or unbranched, saturated or unsaturated Ci -C12 monovalent hydrocarbon chain. In some embodiments, Rio and R20 are each independently H or a branched or unbranched, saturated or unsaturated C3 -C10 monovalent hydrocarbon chain; and Rn and R21 are each independently a branched or unbranched, saturated or unsaturated C3 -C10 monovalent hydrocarbon chain. In some embodiments, Rio and Rn are each independently a linear C4-C8 alkyl; or Rio is H, and Rn is a linear C6-C9 alkyl or C6-C9 alkenyl. In some embodiments, R20 and R21 are each independently a linear C4-C8 alkyl; or R20 is H, and R21 is a linear C6-C9 alkyl or C6-C9 alkenyl. In some embodiments, Rio and Rn are each independently a linear C4-C8 alkyl; or Rio is H, and Rn is a linear C6-C9 alkyl or C6-C9 alkenyl; and R20 and R21 are each independently a linear C4-C8 alkyl; or R20 is H, and R21 is a linear C6-C9 alkyl or C6-C9 alkenyl. In some embodiments, Rio and Rn are each independently a linear C4, Ce, or Cs alkyl; or Rio is H, and Rn is a linear Cs alkyl or Cs alkenyl. In some embodiments, R20 and R21 are each independently a linear C4, Ce, or Cs alkyl; or R20 is H, and R21 is a linear Cs alkyl or Cs alkenyl. In some embodiments, Rio and Rn are each independently a linear C4, Ce, or Cs alkyl; or Rio is H, and Rn is a linear Cs alkyl or Cs alkenyl; and R20 and R21 are each independently a linear C4, Ce, or Cs alkyl; or R20 is H, and R21 is a linear Cs alkyl or Cs alkenyl. Rio and Rn may be the same or different. R20 and R21 may be the same or different. Rio and R20 may be the same or different. Rn and R21 may be the same ordifferent. The group ’Rl1may be the same or different than?Rz1.

[0069] In some embodiments, for the compounds of formula (GI), (GII), (GT), or (GII'), or the compounds of the subgenus formulas under formula (GI), (GII), (GT), or (GIT):Xi and X2 are each independently -C(O)O-, -OC(O)-, -NHC(O)-, or -C(O)NH-;Lio and L20 are each independently a C4-C8 alkylene, optionally substituted by methyl; Ln and L21 are each independently absent or a C1-C3 alkylene;Rio and R20 are each independently H or a branched or unbranched, saturated or unsaturated C3 -C10 monovalent hydrocarbon chain; and / orRn and R21 are each independently a branched or unbranched, saturated or unsaturated C3 -C10 monovalent hydrocarbon chain.In some embodiments, one or more of the following conditions are met:Lio and L20 are each independently a linear C5-C7 alkylene;Ln and L21 are each independently absent or a C2 alkylene;- 23 - LEGAL\113833648\6Rio and Rn are each independently a linear C4-C8 alkyl; or Rio is H, and Rn is a linear C6-C9 alkyl or C6-C9 alkenyl; and / orR20 and R21 are each independently a linear C4-C8 alkyl; or R20 is H, and R21 is a linear C6-C9 alkyl or C6-C9 alkenyl. In some embodiments, one or more of the following conditions are met:Lio and L20 are each independently a linear C5 or C7 alkylene;Ln and L21 are each independently absent or a C2 alkylene;Rio and Rn are each independently a linear C4, Ce, or Cs alkyl; or Rio is H, and Rn is a linear Cs alkyl or Cs alkenyl; and / orR20 and R21 are each independently a linear C4, Ce, or Cs alkyl; or R20 is H, and R21 is a linear Cs alkyl or Cs alkenyl.

[0070] In some embodiments, for the compounds of formula (GI), (GII), (GL), or (GET), or the compounds of the subgenus formulas under formula (GI), (GII), (GL), or (GIL), one or more of the following conditions are met:Xi and X2 are different;Ln and L21 are different;Lio and L20 are different;Rio and Rn are different;R20 and R21 are different; and’Rl1is different than *Rz1.

[0071] In some embodiments, for the compounds of formula (GI), (GII), (GL), or (GII'), or the compounds of the subgenus formulas under formula (GI), (GII), (GL), or (GIL), one of Ln and L21 is absent, and the other one of Ln and L21 is a Ci-Ce alkylene.

[0072] In some embodiments, for the compounds of formula (GI) or (GII), or the compounds of- 24 - LEGAL\113833648\6. Here,_i_represents the connection to Xi or X2.

[0073] In some embodiments, for the compounds of formula (GI) or (GII) or the compounds of L10-X1L-] -]— RR-] NZL20-X2-L21—RR2the subgenus formulas under formula (GI) or (GII), the tail group- 25 - LEGAL\113833648\6LEGAL\113833648\6

[0074] In some embodiments, for the compounds of formula (GI), (GII), (GF), or (GII'), or the compounds of the subgenus formulas under formula (GI), (GII), (GF), or (GIF) (e.g., where RR1and RR2 is for formula (GI) or (GII) or subgenus formulas under- 27 - LEGAL\113833648\6

[0075] In some embodiments, a compound of formula (GI), (GII), (GF), or (GIF), or a compound of the subgenus formulas under formula (GI), (GII), (GF), or (GIF) has the formula of:

[0076] In these formulas, ulO and u20 are each independently 4 to 8; and ul 1 and u21 are each independently 0 to 3. The definitions for the other variables are the same as those described herein for formulas (GI), (GII), (GF), and / or (GIF). In some embodiments, one of ul 1 and u21 is 0, and other one of ul 1 and u21 is 1 to 3. In some embodiments, Rio and Rn are each independently a linear C4-C8 alkyl; or Rio is H or a linear C1-C4 alkyl, and Rn is a linear C6-C9 alkyl or C6-C9 alkenyl. In some embodiments, R20 and R21 are each independently a linear C4-C8 alkyl; or R20 is H or a linear C1-C4 alkyl, and R21 is a linear C6-C9 alkyl or C6-C9 alkenyl.In some embodiments, Rio and Rn are each independently a linear C4-C8 alkyl; or Rio is H or a linear C1-C4 alkyl, and Rn is a linear C6-C9 alkyl or C6-C9 alkenyl; and R20 and R21 are each independently a linear C4-C8 alkyl; or R20 is H or a linear C1-C4 alkyl, and R21 is a linear C6-C9 alkyl or C6-C9 alkenyl. In some embodiments, Rio and Rn are the same. In some embodiments, R20 is H or C1-C4 alkyl, and R21 is a linear C6-C9 alkyl. In some embodiments, Rio and Rn are the- 28 - LEGAL\113833648\6same; and R20 is H or an alkyl that has a shorter chain than R21. In some embodiments, Rio and R11 are the same; and R20 is H or C1-C4 alkyl, and R21 is a linear C6-C9 alkyl.

[0077] In some embodiments, the compound has the formula of:definitions for all the variables are the same as those described herein for formulas (GI), (GII), (GI'), and / or (GII').

[0078] In some embodiments, the compound has the formula of:1). The definitions for all the variables are the same as those described herein for formulas (GI), (GII), (GT), and / or (GIT).

[0079] Non-limiting exemplary compounds for formula (GI), (GII), (GT), or (GIT), or the subgenus formulas under formula (GI), (GII), (GT), or (GIT) are listed in Table 1, Table 2, Table 3, and Table 4 below.Table 1. Exemplary lipid compounds.Lipid LipidStructure StructureNo. No- 29 - LEGAL\113833648\6LEGAL\113833648\6H O25 •™ b0 — o \izO'H H> O) o 27H Hl t z''z- 29ZIQ - ZIH <ZTZTQ / <, X O _ o c00^b0 GO o0 \J / = 31zo o - N)H H33H / ^^yv^^N^bs-Jb.O J OO 0b^F35 ' >oHo J oo IL^ 37^z_ O ZI ^n H N. ZZE _,.„N K o ^O9 o T *N 3ur ^nZCD b b O CD 4143H H45 ^-b O 0 ° bb-ci- 31 - LEGAL\113833648\6LEGAL\113833648\6H69-'A'H HS^S>71xgft^x ft^ XHz ^z73 o > ° ° xAcr / ^ Q2=EQ -? 2 co°? °?75dH O X X X ^ Xo l o o ft 77 ° 1 O O cAO o col X79 ~ )o° / —81^z zo <H / H 0 0 XX XA ^XxX / xxxXx X ~-xx / XXA flN H=cZI Q - 83 zxj O J O Zco< O°?a1 C oo co (f) H 0 0 H O O A b o 85H 0 0xJ 0 ° CI^Zx 87H O O H 0 0 O ° AAF89 / yyX°-yA- 33 - LEGAL\113833648\6LEGAL\113833648\6LEGAL\113833648\6> O ^- <z= D CF-O X OFXH X X-S=O IZ_ _ O _ _ N X J VXJ 1376xyyyx°yxxJ C’H O 0 0X-XX-XXX^XX^O^XXXX^XX^N^ 1,5) \ H [I d 145x~yxy2y^^°'gx^^o oZ^X^XXYXXXXX-N^W VX. 1 (J) \ H M152xyxyzx°yxxxX,CN o o XX N-xx'-'CT xJ, ^, 0 jH154 ^ 1 6 J H x\^^y-'^°y-\xx^0 0 0 0-^X J ouX J H n XX Jxi J o J H n J 156 ZX^u> XXxr X^s^^ X^yxy^’^^0'^^H HX\h O X1O 179 x^x1o > o / XXXyxOyXxJ xyxyxyx^ 2 °o° / ^^'^x'OHHN^186 ^ \zz- xx^^xx / T^x^'xx^ZI°HIZN- NyyIZ188 x^ o X1 0 0CD 5° 0 O O XS o o Xh ^ 1 O JH206 ^ 1 6 J H XXXXXXyX^O^yXxXxlO O o o ^ ^ / ^XXXO-X^X^N^W ^ J AuJ o J H 0 X J H [ V1 J 1^Br 209ux XX^F xxyxyx°yxU0 HHHN^XxXxXQXyx / Xx^^XXX O x1O bb^Br 226xyxyxyx^O^^'^x'x^x\xxyx^°y-xxxx^- 36 - LEGAL\113833648\6o ZI § b0 H O O Ho pO C 00 O O 242 243ZZZ 0 IZ $ 0o=. ° I I IO) Ohrv H) o _^ z249 O J O Br 25060 ° ciAA^^zJ J O Jo( oA A25 II o <3 254 ° °ZI ZI0 0 ZIoo255 0 00 O \ N) 256O Z IZ.0 > O=>§ °6 9 / 257 258 / 00 O 0 o 00-'N-10y'0H IZ247 / ^0 ° 0 ° 0^NH2259^ zH ) O H O 260 / -\0 O 0 ° Cl-"00'F 261 / -^0 o 0 o 00"CF3( OH262 263ZI0 o 264 3230z - \ Table 2, Exemplary lipid compounds.Lipid Structure Lipid StructureNo. No.H285 / U <-> ^ 8 0 287H 0 0 288 289- 37 - LEGAL\113833648\6Hf^il 291 o 6 ooHHo^sAJ 293 JIJ & J & O o ^x^^-x^X^OH H OHH2N^> N P23 / ~x^H OH H OH o o k / F2965s A VFyyyy^o-^x^xj / X^x^^H O—r- > YYoF- ^0 _Brs^H / J N 5 iGN300 — YT^N^V4-O H^COA —;N^^LMCI302 O > O ^ACF3H O O^03°P^7^^°OjO 304 o J O k^FH 1 oo H O O —306^X^X / XJ / ^O. J / ^'^J^H O307 / ~x^H O^ ^^ ^ ^ ^ ^ ^ 308- 38 - LEGAL\113833648\6o c p z — XTable 3. Exemplary lipid compounds (with linker variation).Lipid Structure LipidNo. o / No.265 266J X J. ~ A.. f 1 5 T z ~ ' v J v_ °267 268.,-..> ory -. r ' "" zi.x y -- T"" PJi > - X o269 1 270 H5 co P p271 HN. X p 272 H O O xx^S TQ x.^0 X273 274 H275 v_ v_0° 276 H O O277 278yPZo^^P °279 280 - - J o J I JL H N ■' ' < ■ ', J281 ^ ^ ^ 282 / -J 0 J PxNV^ \J>0283 284xx J o J P / NP _ / yy^X^O^x^^J S" VPTable 4. Exemplary lipid compounds (with tail variation).- 39 - LEGAL\113833648\6LEGAL\113833648\6xpbAX^ QQ j162 AA-XA\ OJL NNXAAA-NA JL AS -AXAXAAAAAAAAN-XAA^' N AAS J pAA^^A 164'A^AA\AOAXAA'AA XX A XX ^ XX A^XXAAA'X^XQXAA'AAXAA^NASA N AAS ACL^AAMAX / X 166AAA,AAAXAAQAAAAA A AASAA AK p ^ A^ l^168 ^d^ XAAXAAAAAAXAXNXXA'XA'' AOkH O O LVpAA AA p L Q 170172A XAAAXATAAAAXXNX^XXA'NAXA^S^AAOh j 175A A A O AA AbXAA ”XAJ''J''A''A'JH 0 0^^^ALLADH 0 0oH^IJXA< AAA-\177AAAAXAGAAAAA XAA N AASA AAXAAAAAAAXAX-HX^'XA"NAASA182^ AVAAAV / X / AAA / A ^ AAS XAXAAA'CAAAAXXNXAAA' N AASCL184- 41 - LEGAL\113833648\6LEGAL\113833648\6p ^0= 0 Hp IZ221 IZ^ zH2230 ( 0H HNP- 228wH H& P O PAgr 230pH O / pwO0IZ 232pIZ0 ^ z H HNP0 / 234( ClH O O P z-ZIrb 3»o

[0080] In some embodiments, the compound has the formula of 0 o(GI-SI). In this formula, the definitions for the variables may be the same as those described herein for formulas (GI), (GII), (GP), (GIF), and / or (GI-TI).

[0081] In some embodiments, in formula (GI-SI), one or more of the variables are defined as follows:RH is aryl or heteroaryl, optionally substituted with one or more of alkyl, hydroxyalkyl, alkoxyalkyl, halo, haloalkyl, -NO2, -C≡N, -ORa, -C(0)NH2, -SO2Ra, or -B(ORa)2, or aryl- 43 - LEGAL\113833648\6optionally fused with a non-aromatic ring;n2 is 0, 1, or 2,Ra and Rd are each independently H or C1-C3 alkyl;A is -CH2-, -CH(OH)-, -C(O)-, -O-, -S-, -NH-, or -NHC(O)-;Bis -C(O)-, oreach of R2and R2', in each occurrence, is independently H, CH3, -NHC(O) CH3; or R2and R2', together with their adjacent carbon atom, form a C3 ring;ulO and u20 are each independently 4 to 8;ul 1 and u21 are each independently 0 to 3;Rio and RH are each independently a linear C4-C8 alkyl; or Rio is H or a linear C1-C4 alkyl, and RH is a linear C6-C9 alkyl or C6-C9 alkenyl; and / orR20 is H or C1-C4 alkyl, and R21 is a linear C6-C9 alkyl.

[0082] In some embodiments, in formula (GI-SI), one of ul 1 and u21 is 0, and other one of ul 1 and u21 is 1 to 3. In some embodiments, the total number of atoms forming the chain connectingRH- to

[0083] In some embodiments, in formula (GI-SI), one or more of the variables are defined as follows:RH is phenyl optionally substituted with one or more of C1-C3 alkyl, aryl, hydroxymethyl, halo, haloalkyl, -NO2, -C≡N, -OCH3, -C(O)NH2, -SO2CH3, or -B(OH)2; dihydrobenzodioxinyl; or tetrazolyl, thiophenyl, pyridinyl, pyrrolyl, pyrazolyl, pyrazinyl, or pyrimidinyl, each optionally substituted with one or more methyl;Ra is H;Rd is H or methyl;nl is 0; and / orn2 is 0 or 2.

[0084] In some embodiments, the head groupn NH- 44 - LEGAL\113833648\6

[0085] The novel lipid compounds disclosed herein, when incorporated into a lipid composition, can confer enhanced delivery to a cell, tissue, or organ, such as kidney, liver, splenic, lymphatic, or marrow - 45 - LEGAL\113833648\6of a subject. In some embodiments, the novel lipid compounds disclosed herein, when incorporated into a lipid composition, can confer preferentially targeting to kidney, liver, splenic, lymphatic, and / or marrow cells, and can thereby confer preferential delivery of a therapeutic agent (e.g., a protein or a nucleic acid) to such cells, tissues, or organs. In some embodiments, the compound is Lipid No. 4, 18, 26, 38, 40, 45, 47, 48, 53, 55, 57, 60, 62, 64, 65, 84, 88, 95, 97, 101, 106, 111, 114, 115, 116, 125, 130, 133, 181, 242, or 243. In some embodiments, the compound is Lipid No. 4, 18, 26, 38, 45, 47, 48, 55, 57, 60, 62, 64, 65, 84, 88, 95, 97, 101, 106, 111, 114, 115, 116, 130, 133, 181, 242, or 243. In some embodiments, the compound is Lipid No. 40, 53, or 125.

[0086] In some embodiments, the compound is Lipid No. 1, 4, 5, 8, 10, 13, 14, 17, 18, 20, 24, 25, 26, 27, 30, 31, 32, 38, 40, 45, 46, 47, 50, 57, 60, 62, 88, 104, 106, 107, 123, 127, 128, 134, 242, or 243. In some embodiments, the compound is Lipid No. 1, 4, 5, 8, 10, 13, 14, 17, 18, 24, 25, 26, 27, 30, 31, 32, 38, 40, 45, 46, 47, 50, 57, 60, 62, 88, 104, 106, 107, 123, 127, 128, 134, 242, or 243. In some embodiments, the compound is Lipid No. 20.

[0087] In some embodiments, the novel lipid compounds disclosed herein, when incorporated into a lipid composition, can confer enhanced delivery to a cell, tissue, or organ, such as kidney.In some embodiments, the novel lipid compounds disclosed herein, when incorporated into a lipid composition, can confer preferentially targeting to a kidney cell or tissue, and can thereby confer preferential delivery of a therapeutic agent (e.g., a protein or a nucleic acid) to a kidney cell or tissue. In some embodiments, the compound is Lipid No. 1, 8, 13, 17, 18, 26, 27, 32, 40, 45, 46, 50, 60, 88, 104, 106, 107, 123, 127, or 134. In some embodiments, the compound is Lipid No.40, 26, 18, 45, 60, or 38.

[0088] In some embodiments, for the compounds of formula (PE-I) or (PE-II), or the compounds of the subgenus formulas under formula (PE-I) or (PE-II), Xio and X20 are each independently -C(O)O- or -OC(O)-; and Xno, Xi2o, X2io, and X220 are each independently absent, -C(O)O-, or -OC(O)-. In some embodiments, Xio or X20 is -C(O)O- (e.g., -C(O)- is connected to Lio or L20, and -O- is connected to Ln or L21). In some embodiments, both Xio and X20 are -C(O)O-. In some embodiments, Xio or X20 are -OC(O)- (e.g., -O- is connected to Lio or L20, and -C(O)- is connected to Ln or L21). In some embodiments, both Xio and X20 are -OC(O)-. In some embodiments, one of Xio and X20 is -C(O)O- and the other is -OC(O)-.

[0089] In the formula (PE-I) or (PE-II) or the subgenus formulas under formula (PE-I) or (PE-II), at least one of Xno, Xi2o, X2io, and X220 is not absent. That is to say, there are at least three ^Lio-Xiio-RinL10-X10-L1 1— <2.v DsNZ? \. V I ^L210-X210-R211L20-A20-L21 —ester groups (-C(O)O- or -OC(O)-) in the tail groupL22o-x22o-R22i [nsome embodiments, Xio and X20 are -C(O)O-; and each of Xno, X120, X210, and X220, if present, is - - 46 - LEGAL\113833648\60C(0)-. In some embodiments, Xio and X20 are -C(O)O-; and each of Xno, Xi2o, X2io, and X220, if present, is -C(O)O-.L110-X-i io-R111Ll20~X120-R121L210-X210-R211L20“A20~L21 —

[0090] In some embodiments, the tail groupL220~x220-R22i contains three ester groups (-C(O)O- or -OC(O)-), wherein Xio and X20 are those described above. In some embodiments, Xio and X20 are -C(O)O-; and (i) Xno and X120 are absent, and (a) one of X210 and X220 is -C(O)O- and the other is absent. In some embodiments, Xio and X20 are -C(O)O-; and (i) Xno and X120 are absent, and (b) one of X210 and X220 is -OC(O)- and the other is absent. In some embodiments, Xio and X20 are -OC(O)-; and (i) Xno and X120 are absent, and (a) one of X210 and X220 is -C(O)O- and the other is absent. In some embodiments, Xio and X20 are -OC(O)-; and (i) Xno and X120 are absent, and (b) one of X210 and X220 is -OC(O)- and the other is absent.L-i 10-X1 io-R111Ll20~X120-R121L21 O-X21 O-R21 1L20“A20~L21 —

[0091] In some embodiments, the tail group1-220 X22°R221contains four ester groups (-C(O)O- or -OC(O)-). Two of Xno, Xi2o, X2io, and X220 are each independently -C(O)O-or -OC(O)-, and the other two are absent. In some embodiments, Xno and X120 are absent. In some embodiments, Xno and X210 are absent. In some embodiments, Xio and X20 are -C(O)O-; and (ii) Xno and X120 are absent, and (a) X210 and X220 are -C(O)O-. In some embodiments, Xio and X20 are -C(O)O-; and (ii) Xno and X120 are absent, and (b) X210 and X220 are -OC(O)-. In some embodiments, Xio and X20 are -OC(O)-; and (ii) Xno and X120 are absent, and (a) X210 and X220 are -C(O)O-. In some embodiments, Xio and X20 are -OC(O)-; and (ii) Xno and X120 are absent, and (b) X210 and X220 are -OC(O)-. In some embodiments, Xio and X20 are -C(O)O-; and Xno and X210 are absent, and X120 and X220 are -C(O)O-. In some embodiments, Xio and X20 are -C(O)O-; and Xno and X210 are absent, and X120 and X220 are -OC(O)-. In some embodiments, Xio and X20 are -OC(O)-; and Xno and X210 are absent, and X120 and X220 are -C(O)O-. In some embodiments, Xio and X20 are -OC(O)-; and Xno and X210 are absent, and X120 and X220 are -OC(O)-.^L1 10-X1 10-R.|.|.|L10-X10-L1 1—S_NZL120-A120-K121? \ v i ^L210-X210-R211L20-A20-L21 - \

[0092] In some embodiments, the tail group1-220 X22°R221contains five ester groups (-C(O)O- or -OC(O)-). Three of Xno, Xi2o, X2io, and X220 are each independently -C(O)O-or -OC(O)-, and the other one is absent. In some embodiments, Xio and X20 are -C(O)O-; and (iii) one of Xno and X120 is absent, and (a) the other is -C(O)O-; and X210 and X220 are -C(O)O-.- 47 - LEGAL\113833648\6In some embodiments, Xio and X20 are -C(O)O-; and (iii) one of Xno and X120 is absent, and (a) the other is -OC(O)-; and X210 and X220 are -OC(O)-. In some embodiments, Xio and X20 are -OC(O)-; and (iii) one of Xno and X120 is absent, and (a) the other is -C(O)O-; and X210 and X220 are -C(O)O-. In some embodiments, Xio and X20 are -OC(O)-; and (iii) one of Xno and X120 is absent, and (a) the other is -OC(O)-; and X210 and X220 are -OC(O)-.^L1 10-X1 10-R.|.|.|L10-X10-L1 1— tNZ 20-? \ v i ^L210-X210-R211L20-A20-L21 - \

[0093] In some embodiments, the tail group1-220 X22°R221contains six ester groups (-C(O)O- or -OC(O)-). All four of Xno, Xi2o, X2io, and X220 are each independently -C(O)O- or -OC(O)-. In some embodiments, Xio and X20 are -C(O)O-; and Xno, Xi2o, X2io, and X220 are -C(O)O-. In some embodiments, Xio and X20 are -C(O)O-; and Xno, Xi2o, X2io, and X220 are -OC(O)-. In some embodiments, Xio and X20 are -OC(O)-; and Xno, Xi2o, X2io, and X220 are -C(O)O-. In some embodiments, Xio and X20 are -OC(O)-; and Xno, Xi2o, X2io, and X220 are -OC(O)-.

[0094] In some embodiments, for the compounds of formula (PE-I) or (PE-II), or the compounds of the subgenus formulas under formula (PE-I) or (PE-II), Lio and L20 are each independently a C1-C12 alkylene, optionally substituted by C1-C3 alkyl. In some embodiments, Lio and L20 are each independently a C4-C8 alkylene, optionally substituted by methyl. In some embodiments, Lio and L20 are each independently a linear C5-C7 alkylene. In some embodiments, Lio and L20 are each independently a linear C5 or C7 alkylene. Lio and L20 may be the same or different.

[0095] In some embodiments, for the compounds of formula (PE-I) or (PE-II), or the compounds of the subgenus formulas under formula (PE-I) or (PE-II), Ln, L21, Luo, L120, L210, and L220 are each independently absent or a Ci-Ce alkylene. In some embodiments, Ln, L21, Lno, L120, L210, and L220 are each independently absent or a C1-C3 alkylene. In some embodiments, Ln, L21, Lno, L120, L210, and L220 are each independently absent or a C1-C2 alkylene. Ln and L21 may be the same or different. Lno and L120 may be the same or different. L210, and L220 may be the same or different. Lno and L210 may be the same or different. L120 and L220 may be the same or different.

[0096] In some embodiments, for the compounds of formula (PE-I) or (PE-II), or the compounds of the subgenus formulas under formula (PE-I) or (PE-II), Rm, R121, R211, and R221 each are independently a branched or unbranched, saturated or unsaturated Ci -C12 monovalent hydrocarbon chain. In some embodiments, Rm, R121, R211, and R221 are each independently H or branched or unbranched, saturated or unsaturated Ci -C10 monovalent hydrocarbon chain.

[0097] In some embodiments, for the compounds of formula (PE-I) or (PE-II), or the- 48 - LEGAL\113833648\6compounds of the subgenus formulas under formula (PE -I) or (PE-II):Lio and L20 are each independently a C4-C8 alkylene, optionally substituted by methyl; Ln, L21, Luo, L120, L210, and L220 are each independently absent or a C1-C3 alkylene; and / orR111, R121, R211, and R221 are each independently H or branched or unbranched, saturated or unsaturated Ci -C10 monovalent hydrocarbon chain.

[0098] In some embodiments, for the compounds of formula (PE-I) or (PE-II), or the compounds of the subgenus formulas under formula (PE -I) or (PE-II):Lio and L20 are each independently a linear C5-C7 alkylene; and / orLn, L21, Luo, L120, L210, and L220 are each independently absent or a C1-C2 alkylene.

[0099] In some embodiments, for the compounds of formula (PE-I) or (PE-II), or the compounds of the subgenus formulas under formula (PE -I) or (PE-II), in each of the tail groups,

[0100] In some embodiments, for the compounds of formula (PE -I) or (PE-II), or the compounds of the subgenus formulas under formula (PE -I) or (PE-II), the tail groupL10-X10— L11 - < L| v D_S_NZ? \. v i ^L21Q—X210_R211L220-X220-R221maybe:- 49 - LEGAL\113833648\6connection to LL.

[0101] Non-limiting exemplary compounds for formula (PE -I) or (PE -II), or the subgenus formulas under formula (PE-I) or (PE-II) are listed in Table 5.Table 5. Exemplary lipid compounds (with polyester tail).Lipid LipidStructure StructureNo. No159 241i n? r? ° L^ IiLi J °L N r °00 HOO H— 174 322 O A Oo ' ° r'''"''""'00 H- 50 - LEGAL\113833648\6LEGAL\113833648\6

[0102] In some embodiments, a compound of formula (PE-I) or (PE -II), or a compound of the subgenus formulas under formula (PE-I) or (PE -II) has the formula of:R2R2Ra1 v 1 ^'*-110-X110-R111\ / n9 I. MO-X10-L11 x p N _NZL120-A120-K121M\ \| Y I ^ LL21O_ AX210- RK211 I]L20-A20_ L21 o L220— X220-R221(PEJ-S!)IN THLSformula, the definitions for the variables may be the same as those described herein for formulas (PE-I) and / or (PE-II).

[0103] In some embodiments, in (PE -LSI), one or more of the variables are defined as follows:RH is alkyl, aryl, or heteroaryl, optionally substituted with one or more of alkyl, hydroxyalkyl, alkoxyalkyl, halo, haloalkyl, -NO2, -C≡N, -ORa, -C(0)NH2, -SChRd, or -B(ORa)2, or aryl optionally fused with a non-aromatic ring;n2 is 0, 1, or 2,Ra and Rd are each independently H or C1-C3 alkyl;O OAis -CH2-, -CH(OH)-, -C(O)-, -O-, -S-,, -NH-, or -NHC(O)-; and / or each of R2 and R2', in each occurrence, is independently H, CH3, -NHC(O)CH3; or R2 and R2', together with their adjacent carbon atom, form a C3 ring.

[0104] In some embodiments, the total number of atoms forming the chain connecting RH- toL1o —— NzL2°—is nine.

[0105] In some embodiments, in (PE-I-SI), one or more of the variables are defined as- 52 - LEGAL\113833648\6follows:RH is methyl or phenyl optionally substituted with one or more of C1-C3 alkyl, aryl, hydroxymethyl, halo, haloalkyl, -NO2, -C≡N, -OCH3, -C(0)NH2, -SO2CH3, or -B(0H)2; dihydrobenzodioxinyl; or tetrazolyl, thiophenyl, pyridinyl, pyrrolyl, pyrazolyl, pyrazinyl, or pyrimidinyl, each optionally substituted with one or more methyl groups;Ra is H;Rd is H or methyl;nl is 0; and / orn2 is 0 or 2.

[0106] In some embodiments, the head groupowherein I represents the connection to LL.

[0107] The novel lipid compounds disclosed herein, when incorporated into a lipid composition, can confer enhanced delivery to a cell, tissue, or organ, such as kidney, liver, splenic, lymphatic, or marrow of a subject. In some embodiments, the novel lipid compounds disclosed herein, when incorporated into a lipid composition, can confer preferentially targeting to kidney, liver, splenic, lymphatic, and / or marrow cells, and can thereby confer preferential delivery of a therapeutic agent (e.g., a protein or a nucleic acid) to such cells, tissues, or organs. In some embodiments, the compound is Lipid No. 159, 224, 240, 237, 225, 238, 239, 208, 215, 217, or 174. In some embodiments, the compound is Lipid No. 159, 240, 237, 225, 238, 239, 208, 215, 217, or 174.Definition

[0108] The disclosures of each patent, patent application, and publication cited or described in this document are hereby incorporated herein by reference, in their entirety.

[0109] Unless defined otherwise, all technical and scientific terms used herein have the same meaning as is commonly understood by one of skill in the art to which this disclosure belongs.

[0110] In the present disclosure the singular forms “a”, “an”, and “the” include the plural reference, and reference to a particular numerical value includes at least that particular value,- 53 - LEGAL\113833648\6unless the context clearly indicates otherwise. Thus, for example, a reference to “a compound” is a reference to one or more of such compounds and equivalents thereof known to those skilled in the art, and so forth. Furthermore, when indicating that a certain chemical moiety “may be” X, Y, or Z, it is not necessarily intended by such usage to exclude other choices for the moiety; for example, a statement to the effect that Ri “may be alkyl, aryl, or amino” does not necessarily exclude other choices for Ri, such as halo, aralkyl, and the like.

[0111] When values are expressed as approximations, by use of the antecedent “about,” it will be understood that the particular value forms another embodiment. As used herein, “about X” (where X is a numerical value) preferably refers to ±10% of the recited value, inclusive. For example, the phrase “about 8” may refer to a value of 7.2 to 8.8, inclusive; as another example, the phrase “about 8%” may refer to a value of 7.2% to 8.8%, inclusive. Also, when the term “about” precedes a range, it is understood that the term modifies both recited endpoints and all points embraced within the range. For example, the phrase “about 1-10” is understood to mean “about 1 to about 10”, as well as “about x”, wherein x refers to any value between 1 and 10. Where present, all ranges are inclusive and combinable. For example, when a range of “1 to 5” is recited, the recited range should be construed as including ranges “1 to 4”, “1 to 3”, “1-2”, “1-2 & 4-5”, “1-3 & 5”, and the like.

[0112] When a list is presented, unless stated otherwise, it is to be understood that each individual element and every combination is to be interpreted as separate embodiments. For example, a list of embodiments presented as “A, B, or C” is to be interpreted as including the embodiments, “A,” “B,” “C,” “A or B,” “A or C,” “B or C,” or “A, B, or C.”

[0113] Protective groups are abbreviated according to the system disclosed in Greene, T. W. and Wuts, P. G. M., Protective Groups in Organic Synthesis 2d. Ed., Wiley & Sons, 1991, which is incorporated in its entirety herein. For example, “CBZ” or “Cbz” or “Z” stands for carbobenzyloxy or benzyloxycarbonyl, “Boc” or “BOC” represents / -butoxycarbonyl, “Alloc” denotes allyloxycarbonyl, Bz means benzoyl, and “Fmoc” stands for 9-fluorenylmethoxycarbonyl.

[0114] As used herein, the term “compound” is meant to include all the isomers and isotopes of the structure depicted, all the pharmaceutically acceptable salts, solvates, or hydrates thereof, and all crystal forms (e.g., crystal polymorphs), crystal form mixtures, or anhydrides or hydrates thereof.

[0115] The term “isotope” refers to atoms having the same atomic number but different mass numbers resulting from a different number of neutrons in the nuclei. For example, isotopes of hydrogen include tritium (3H) and deuterium (2H).

[0116] A "stereoisomer" refers to a compound made up of the same atoms bonded by the same- 54 - LEGAL\113833648\6bonds but having different three-dimensional structures, which are not interchangeable. The present disclosure contemplates various stereoisomers and mixtures thereof and includes "enantiomers", which refers to two stereoisomers whose molecules are non-superimposable mirror images of one another.

[0117] “ Isomers.” The compounds described herein or their pharmaceutically acceptable salts may include all isomers, such as geometrical isomers, optical isomers based on an asymmetrical carbon, stereoisomers, tautomers, and the like. For instance, the compounds can contain one or more stereocenters and may thus give rise to geometic isomers (e.g., double bond causing geometric E / Z isomers), enantiomers, diastereomers (e.g., enantiomers (i.e., (+) or (-)) or cis / trans isomers), and other stereoisomeric configurations that may be defined, in terms of absolute stereochemistry, as (R)- or (S)- such as for sugar anomers, or as (D)- or (L)- such as for amino acids. The present disclosure is meant to include all such possible isomers, as well as their racemic and optically pure forms. Optically active (+) and (-), (R)- and (S)-, or (D)- and (L)-isomers may be prepared using chiral synthons or chiral reagents, or resolved using conventional techniques, for example, chromatography and fractional crystallization. Conventional techniques for the preparation / isolation of individual enantiomers include chiral synthesis from a suitable optically pure precursor or resolution of the racemate (or the racemate of a salt or derivative) using, for example, chiral high pressure liquid chromatography (HPLC). Enantiomeric and stereomeric mixtures of compounds and means of resolving them into their component enantiomers or stereoisomers are well-known. When the compounds described herein contain ol efini c double bonds or other centers of geometric asymmetry, and unless specified otherwise, it is intended that the compounds include both E and Z geometric isomers. Likewise, all tautomeric forms are also intended to be included.

[0118] The term “crystal polymorphs”, “polymorphs” or “crystal forms” means crystal structures in which a compound (or a salt or solvate thereof) can crystallize in different crystal packing arrangements, all of which have the same elemental composition. Different crystal forms usually have different X-ray diffraction patterns, infrared spectral, melting points, density hardness, crystal shape, optical and electrical properties, stability and solubility. Recrystallization solvent, rate of crystallization, storage temperature, and other factors may cause one crystal form to dominate. Crystal polymorphs of the compounds can be prepared by crystallization under different conditions.

[0119] Crystallization of the compounds disclosed herein may produce a solvate. As used herein, the term “solvate” refers to an aggregate that compri ses one or more molecules of an ionizable lipid of the disclosure with one or more molecules of solvent. The solvent may be water, m which case the solvate may be a hydrate, including a monohydrate, dihydrate,- 55 - LEGAL\113833648\6hemihydrate, sesquihydrate, trihydrate, tetrahydrate and the like. Alternatively, the solvent may¬ be an organic solvent. Solvates of the compound may be true solvates, while in other cases, the compound may merely retain adventitious water or be a mixture of water plus some adventitious solvent.

[0120] As used herein, the term “pharmaceutically acceptable carrier” preferably refers to a material that can be incorporated into a composition and administered to a patient without causing unacceptable biological effects or interacting in an unacceptable manner with other components of the composition. Such pharmaceutically acceptable materials typically have met the required standards of toxicological and manufacturing testing, and include those materials identified as suitable inactive ingredients by the U. S. Food and Drug Administration.

[0121] The term “pharmaceutically acceptable salt” include the salts derived from suitable inorganic and organic acids and bases. Examples of pharmaceutically acceptable, nontoxic acid addition salts are 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 include adipate, alginate, ascorbate, aspartate, benzenesulfonate, benzoate, bisulfate, borate, butyrate, camphorate, camphorsulfonate, citrate, cyclopentanepropionate, digluconate, dodecylsulfate, ethanesulfonate, formate, fumarate, glucoheptonate, glycerophosphate, gluconate, hemisulfate, heptanoate, hexanoate, hydroiodide, 2-hydroxy-ethanesulfonate, lactobionate, lactate, laurate, lauryl sulfate, malate, maleate, malonate, methanesulfonate, 2-naphthalenesulfonate, nicotinate, nitrate, oleate, oxalate, palmitate, pamoate, pectinate, persulfate, 3- phenylpropionate, phosphate, picrate, pivalate, propionate, stearate, succinate, sulfate, tartrate, thiocyanate, p-toluenesulfonate, undecanoate, valerate salts, and the like. Salts derived from appropriate bases include alkali metal, alkaline earth metal, ammonium and N+(CI alkylAsalts. Representative alkali or alkaline earth metal salts include sodium, lithium, potassium, calcium, magnesium, and the like. Further pharmaceutically acceptable salts include, when appropriate, nontoxic ammonium, quaternary ammonium, and amine cations formed using counterions such as halide, hydroxide, carboxylate, sulfate, phosphate, nitrate, sulfonate and aryl sulfonate. Further pharmaceutically acceptable salts include salts formed from the quartemization of an amine using an appropriate electrophile, e.g., an alkyl halide, to form a quartemized alkylated amino salt.

[0122] An “effective amount” or “therapeutically effective amount” of a therapeutic agent such as a therapeutic nucleic acid is an amount sufficient to produce the desired effect, e.g., an increase or inhibition of expression of a target sequence in comparison to the normal expression level detected in the absence of the nucleic acid. Suitable assays for measuring expression of a- 56 - LEGAL\113833648\6target gene or target sequence include, e.g., examination of protein or RNA levels using techniques known to those of skill in the art such as dot blots, northern blots, in situ hybridization, ELISA, immunoprecipitation, enzyme function, fluorescence or luminescence of suitable reporter proteins, as well as phenotypic assays known to those of skill in the art.

[0123] “ Treating” or “treatment” as used herein covers the treatment of the disease or disorder in a mammal, such as a human, having the disease or disorder, and includes:(i) preventing the disease or disorder from occurring in a mammal, in particular, when such mammal is predisposed to the condition but has not yet been diagnosed as having it;(ii) inhibiting the disease or disorder, i.e., arresting its development;(lii) relieving the disease or disorder, i.e., causing regression of the disease or disorder; or (iv) relieving the symptoms resulting from the disease or disorder, i.e., relieving pain without addressing the underlying disease or condition. As used herein, the terms “disease” and “disorder” may be used interchangeably or may be different in that the particular malady or or disorder may not have a known causative agent (so that etiology has not yet been worked out) and it is therefore not yet recognized as a disease but only as an undesirable condition or syndrome, wherein a more or less specific set of symptoms have been identified by clinicians.

[0124] As used herein, “ionizable lipid” refers to a lipid that can be ionized, e.g., dissociated to produce one or more electrically charged species, under a given condition (e.g., pH).

[0125] The term “neutral lipid” refers to any of a lipid that exists either in an uncharged or neutral zwitterionic form at a selected pH.

[0126] The term “liposome” as used herein refers to a composition comprising an outer lipid layer membrane (e.g., a single lipid bi-layer known as unilamellar liposomes or multiple lipid bilayers known as multilamellar liposomes) surrounding an internal aqueous space which may contain a cargo. See, e.g., Cullis et ah, Biochim. Biophys Acta, 559: 399-420 (1987), which is incorporated herein by reference in its entirety. A unilamellar liposome generally has a diameter in the range of about 20 to about 400 nanometers (nm), about 50 to about 300 nm, about 100 to about 200 nm, or about 300 to about 400 nm. A multilamellar liposome usually has a diameter in the range of about 1 to about 10 pm and may comprise anywhere from 2 to hundreds of concentric lipid bilayers alternating with layers of an aqueous phase.

[0127] The term “lipid nanoparticle” refers to a particle having at least one dimension on the order of nanometers (e.g., 1-1,000 nm) and comprising one or more lipids (e.g., the lipid compounds described herein, and a second lipid (e.g., cationic, anionic, ionizable, or zwitterionic lipid)) for encapsulation of a therapeutic agent. LNPs may also include neutral lipids such as phospholipid molecules belonging to the phosphatidylcholine (PC) class; sterols, such as cholesterol; and polyethylene glycol (PEG). LNPs may be taken up by cells \ia endocytosis and- 57 - LEGAL\113833648\6the ionizability of the lipids at low pH enables endosomal escape, which can allow release of cargo into the cytoplasm. LNPs are liposome-like structures. Exemplary lipid nanoparticle composition are formulations of ionizable lipids, sterols (or hydrophobic molecules), structural lipids such as phospholipids, polyethyleneglycol (PEG) lipids, and potentially additional components (see Nature Nanotechnology 15:313-320 (2020), which is incorporated herein by reference in its entirety'), or single molecules containing combinations of ionizable lipid, sterol, structural phospholipid, and shielding groups (see Nature Materials 20:701-710 (2021), which is incorporated herein by reference in its entirety). These components may be mixed with a therapeutic agent (e.g., nucleic acid molecules such as mRNA or circRNA) to be formulated into LNP composition.

[0128] As used herein, the term “size” refers to the hydrodynamic diameter of a lipid nanoparticle population. The measurement of the size of a lipid composition may be used to indicate the size and population distribution (polydispersity index, PDI) of the composition.

[0129] As used herein, the “polydispersity index” is a ratio between weight-average molar mass and Mn is the number- verage molar mass that describes the homogeneity' of the particle size distribution of a system. A small value, e.g., less than 0.3, indicates a narrow particle size distribution.

[0130] As used herein, the term “apparent pKa” refers to the pH at which 50% of the lipid composition (e.g., LNP) is protonated. This can be used as an indicator of the pH range that the lipid composition (e.g., LNP) will be protonated, and thus initiate the endosomal escape process in a nucleotide delivery'.

[0131] As used herein, the term “zeta potential” refers to the electrokinetic potential of lipid, e.g., in a lipid composition (e.g., a LNP composition). The zeta potential may describe the surface charge of a LNP composition. Zeta potential is useful in predicting organ tropism and potential interaction with serum proteins.

[0132] As used herein, “encapsulated” by a lipid refers a therapeutic agent, such as a nucleic acid (e.g., mRNA), that is fully or partially encapsulated to by a lipid composition (e.g., liposome or LNP). In some embodiments, nucleic acid (e.g., mRNA) is fully encapsulated in a lipid composition (e.g., liposome or LNP).

[0133] As used herein, “encapsulation efficiency” or “entrapment efficiency” refers to the percentage of an encapsulated cargo (e.g., a therapeutic agent) that is successfully incorporated into (e.g., encapsulated or otherwise associated with) the lipid composition (e.g., a LNP or liposome), relative to the ini tial total amount of therapeutic agent provided. For example, if 97 mg of a therapeutic agent are encapsulated in a lipid composition out of a total 100 mg of the therapeutic agent initially provided, the encapsulation efficiency may be given as 97%.- 58 - LEGAL\113833648\6Encapsulation efficiency can be used to indicate the efficiency of an encapsulated cargo (e.g., a nucleic acid molecule) loading into the lipid composition using a particular formulation method and formulation recipe.

[0134] The term “alkyl” refers to a hydrocarbon chain that may be a straight chain or branched chain, containing the indicated number of carbon atoms. For example, C1-C12 alkyl indicates that the group may have from 1 to 12 (inclusive) carbon atoms in it. Unless otherwise indicated, “alkyl” generally refers to C1-C24 alkyl (e.g., C1-C15 alkyl, C1-C12 alkyl, Ci-Cs alkyl, Ci-Ce alkyl, C1-C4 alkyl, or C1-C3 alkyl). Unless stated otherwise specifically in the specification, an alkyl group is optionally substituted.

[0135] The term "alkenyl" refers to a straight or branched hydrocarbon chain containing 2-8 carbon atoms and characterized in having one or more double bonds. Unless otherwise indicated, “alkenyl” generally refers to C2-C8 alkenyl (e.g., C2-C6 alkenyl, C2-C4 alkenyl, or C2-C3 alkenyl). Examples of a typical alkenyl include, but not limited to, allyl, propenyl, 2-butenyl, 3 -hexenyl and 3 -octenyl groups.

[0136] The term "alkynyl" refers to a straight or branched hydrocarbon chain and characterized in having one or more triple bonds. Unless otherwise indicated, “alkynyl” generally refers to C2-C8 alkynyl (e.g., C2-C6 alkynyl, C2-C4 alkynyl, or C2-C3 alkynyl). Some examples of typical alkynyl groups are ethynyl, 2-propynyl, and 3-methylbutynyl, and propargyl. The sp2and sp3carbons may optionally serve as the point of attachment of the alkenyl and alkynyl groups, respectively.

[0137] The term “alkoxy” refers to an -O-alkyl radical.

[0138] The term “thioalkoxy” refers to an -S-alkyl radical.

[0139] The term “alkylene” refers to a bivalent form of an alkyl group. An “alkylene chain” is a polymethylene group, i.e., — (CH2)n—, wherein n is a positive integer, preferably from 1 to 6, from 1 to 4, from 1 to 3, from 1 to 2, or from 2 to 3. A substituted alkylene chain is a polymethylene group in which one or more methylene hydrogen atoms are replaced with a substituent. Suitable substituents include those described below.

[0140] The term “alkenylene” refers to a bivalent alkenyl group. A substituted alkenylene chain is a polymethylene group containing at least one double bond in which one or more hydrogen atoms are replaced with a substituent. Suitable substituents include those described below.

[0141] The term “alkynylene” refers to a divalent form of an alkynyl group. The sp2and sp3carbons may optionally serve as the point of attachment of the alkynylene groups. Examples of alkynlene groups include ethynylene, propynylene, n-butynylene, and the like.

[0142] The term “aryl” refers to a 6-carbon monocyclic or 10-carbon bicyclic aromatic ring- 59 - LEGAL\113833648\6system wherein 0, 1, 2, 3, or 4 atoms of each ring may be substituted by a substituent. The term “aryl” may be used interchangeably with the term “aryl ring.” Examples of aryl groups include phenyl, biphenyl, naphthyl, anthracyl, and the like, which may bear one or more substituents. Also included within the scope of the term “aryl,” as it is used herein, is a group in which an aromatic ring is fused to one or more non-aromatic rings, such as indanyl, phthalimidyl, naphthimidyl, phenanthridinyl, or tetrahydronaphthyl, and the like.

[0143] The term “arylalkyl” or the term “aralkyl” refers to alkyl substituted with an aryl. The term “arylalkenyl” refers to alkenyl substituted with an aryl. The term “arylalkynyl” refers to alkynyl substituted with an aryl.

[0144] The term “heteroarylalkyl” refers to alkyl substituted with a heteroaryl. The term “heteroarylalkenyl” refers to alkenyl substituted with a heteroaryl. The term “heteroarylalkynyl” refers to alkynyl substituted with a heteroaryl.

[0145] The term “arylalkoxy” or “aralkoxy” refers to an alkoxy substituted with aryl.

[0146] The term “cycloalkyl” or “cyclyl” as employed herein includes saturated and partially unsaturated, but not aromatic, cyclic hydrocarbon groups having 3 to 12 carbons, for example, 3 to 8 carbons, and, for example, 3 to 6 carbons, wherein the cycloalkyl group additionally may be optionally substituted. Cycloalkyl groups include, without limitation, cyclopropyl, cyclobutyl, cyclopentyl, cyclopentenyl, cyclohexyl, cyclohexenyl, cycloheptyl, and cyclooctyl.

[0147] The cyclic group includes monocyclic and bicyclic group (e.g., bridged cycle, fused cycle, or spiro). A fused cycle, e.g., a fused bicyclic is a cyclic group having two rings sharing two adjacent carbon atoms and one bond. An exemplary fused bicyclic is decalin (bicyclodecane). A bridged cycle is a cyclic group containing two or more rings sharing nonadj acent “bridgehead” carbon atoms, that is to say, the two bridgehead carbons are separated by “bridges” containing at least one carbon. An exemplary bridged bicyclic is norbomane (bicycloheptane). A spiro cycle is a cyclic group containing two rings connecting at a single quaternary carbon atom. The two rings do not share a bond, only a carbon atom. An exemplary spiro bicyclic is spiro[4.5]decane.

[0148] The term “halo” or “halogen” refers to any radical of fluorine, chlorine, bromine or iodine.

[0149] The term “heteroaryl” or “heteroar-” refers to an aromatic 5-8 membered monocyclic, 8-12 membered bicyclic, or 11-14 membered tricyclic ring system having 1-3 heteroatoms if monocyclic, 1-6 heteroatoms if bicyclic, or 1-9 heteroatoms if tricyclic, said heteroatoms selected from O, N, or S (e.g., carbon atoms and 1-3, 1-6, or 1-9 heteroatoms of N, O, or S if monocyclic, bicyclic, or tricyclic, respectively), wherein 0, 1, 2, 3, or 4 atoms of each ring may be substituted by a substituent. The term also include groups in which a heteroaromatic ring is fused to one or- 60 - LEGAL\113833648\6more aryl, cycloalkyl, or heterocyclyl rings, where the radical or point of attachment is on the heteroaromatic ring. Examples of heteroaryl groups include pyrrolyl, pyridyl, pyridazinyl, pyrazolyl, triazolyl, tetrazolyl, oxazolyl, isoxazolyl, oxadiazolyl, furanyl, imidazolyl, benzimidazolyl, pyrimidinyl, pyrazinyl, indolizinyl, thiophenyl or thienyl, quinolinyl, indolyl, thiazolyl, isothiazolyl, thiadiazolyl, purinyl, naphthyridinyl, pteridinyl, isoindolyl, benzothienyl, benzofuranyl, dibenzofuranyl, indazolyl, benzthiazolyl, quinolyl, isoquinolyl, cinnolinyl, phthalazinyl, quinazolinyl, quinoxalinyl, 4H-quinolizinyl, carbazolyl, acridinyl, phenazinyl, phenothiazinyl, phenoxazinyl, tetrahydroquinolinyl, tetrahydroisoquinolinyl, and pyrido[2,3-b]-l,4-oxazin-3(4H)-one and the like.

[0150] The term “heterocyclyl,” “heterocycle,” “heterocyclic radical,” or “heterocyclic ring” refers to a nonaromatic 5-8 membered monocyclic, 8-12 membered bicyclic, or 11-14 membered tricyclic ring system having 1-3 heteroatoms if monocyclic, 1-6 heteroatoms if bicyclic, or 1-9 heteroatoms if tricyclic, said heteroatoms selected from O, N, or S (e.g., carbon atoms and 1-3, 1-6, or 1-9 heteroatoms of N, O, or S if monocyclic, bicyclic, or tricyclic, respectively), wherein 0, 1, 2 or 3 atoms of each ring may be substituted by a substituent. When used in reference to a ring atom of a heterocycle, the term “nitrogen” includes a substituted nitrogen. As an example, in a saturated or partially unsaturated ring having 0-3 heteroatoms selected from oxygen, sulfur or nitrogen, the nitrogen may be N (as in 3,4-dihydro-2H-pyrrolyl), NH (as in pyrrolidinyl), or +NR (as in N-substituted pyrrolidinyl). Examples of heterocyclyl groups include trizolyl, tetrazolyl, piperazinyl, pyrrolidinyl, dioxanyl, dioxolanyl, diazepinyl, oxazepinyl, thiazepinyl, morpholinyl, tetrahydrofuranyl, tetrahydrothiophenyl pyrrolidinyl, piperidinyl, pyrrolinyl, tetrahydroquinolinyl, tetrahydroisoquinolinyl, decahydroquinolinyl, oxazolidinyl, quinuclidinyl, and the like.

[0151] The term “heterocyclylalkyl” refers to an alkyl group substituted by a heterocyclyl, wherein the alkyl and heterocyclyl portions independently are optionally substituted.

[0152] A divalent radical of an alkyl, alkenyl, aryl, heteroaryl, cycloalkyl, heterocyclyl, or bridged cyclyl is formed by removal of a hydrogen atom from an alkyl, alkenyl, aryl, heteroaryl, cycloalkyl, heterocyclyl, and bridged cyclyl radical, respectively (or by removal of two hydrogen atoms from an alkane, alkene, arene, heteroarene, cycloalkane, heterocycle, or bridged cycle respectively). A divalent radical of an aryl is also referred to herein as “arylene,” and a divalent radical of a heteroaryl is also referred to herein as “heteroarylene.”

[0153] The term “alkoxyalkyl” refers to an alkyl substituted with an alkoxy group.

[0154] The term “hydroxyalkyl” refers to an alkyl substituted with a hydroxy group.

[0155] The term “alkylthio” refers to an S-alkyl radical.

[0156] The term “arylthio” refers to an S-aryl radical.- 61 - LEGAL\113833648\6

[0157] The term “aryloxy” refers to an O-aryl radical.

[0158] The term “arylamino” refers to an amino-aryl radical.

[0159] The term “alkylsulfonyl” refers to a -SO₂-alkyl radical.

[0160] The term “haloalkyl” refers to an alkyl group in which one or more hydrogens are replaced by a halogen.

[0161] The term “oxo” refers to an oxygen atom, which forms a carbonyl when attached to carbon, an N-oxide when attached to nitrogen, and a sulfoxide or sulfone when attached to sulfur.

[0162] The term “acyl” refers to an alkylcarbonyl, cycloalkylcarbonyl, arylcarbonyl, heterocyclylcarbonyl, or heteroarylcarbonyl substituent, any of which may be further substituted by substituents.

[0163] The term “aminoalkyl” refers to an alkyl substituted with an amino.

[0164] The term “alkylamino” refers to an amino substituted with an alkyl.

[0165] The term “aminocarbonyl” refers to an -C(O)-amino radical.

[0166] The term "substituted" refers to the replacement of one or more hydrogen radicals in a given structure with the radical of a specified substituent including, but not limited to: halo, alkyl, alkenyl, alkynyl, aryl, heterocyclyl, thiol, alkylthio, arylthio, alkylthioalkyl, arylthioalkyl, alkylsulfonyl, alkylsulfonylalkyl, arylsulfonylalkyl, alkoxy, aryloxy, aralkoxy, aminocarbonyl, alkylaminocarbonyl, arylaminocarbonyl, alkoxycarbonyl, aryloxycarbonyl, haloalkyl, amino, trifluoromethyl, cyano, nitro, alkylamino, arylamino, alkylaminoalkyl, arylaminoalkyl, aminoalkylamino, hydroxy, alkoxyalkyl, carboxyalkyl, alkoxycarbonylalkyl, aminocarbonylalkyl, acyl, aralkoxycarbonyl, carboxylic acid, sulfonic acid, sulfonyl, phosphonic acid, aryl, heteroaryl, heterocyclic, and aliphatic. It is understood that the substituent can be further substituted.Lipid Compositions and Preparation

[0167] The lipid compounds disclosed herein may be used to form a lipid composition (e.g., liposome or lipid nanoparticle). Thus, another aspect of the invention relates to a lipid composition comprising a lipid component and the compounds as described herein in the above aspects relating to the lipid compounds. Suitable compounds are all the lipid compounds described herein, including all the compounds of formulas (GI), (GII), (GT), (GIT), (PE-I), and / or (PE -II) and any of the compounds belonging to any subgenus formulas or species under formula (GI), (GII), (GT), (GIT), (PE-I), or (PE-II), described herein.

[0168] As described herein, suitable compounds to be used in the lipid composition include all the isomers and isotopes of the lipid compounds described above, as well as all the pharmaceutically acceptable salts, solvates, or hydrates thereof, and all crystal forms, crystal form mixtures, and anhydrides or hydrates.- 62 - LEGAL\113833648\6

[0169] All above descriptions and all embodiments discussed in the above aspects relating to the aspects of the lipid compounds, including the compounds covered by formulas (GI), (GII), (GI'), (GIF), (PE -I), and (PE-II), and the subgenus formulas and species under formula (GI), (GII), (GP), (GIF), (PE-I), and (PE-II), including various variables in these formulas, are all applicable to these aspects of the invention relating to the lipid composition (e.g., liposome or lipid nanoparticle).

[0170] In some embodiments, the lipid composition is in the form of a liposome. In some embodiments, the lipid composition is in the form of a lipid nanoparticle (LNP).

[0171] As described above, the lipid composition comprises one or more other lipid components, besides the novel lipid compound disclosed herein. In some embodiments, the other lipid component(s) may be cationic, non-cationic (e.g., neutral, anionic, or zwitterionic), or ionizable. In some embodiments, the other lipid component(s) in the lipid composition may be a cationic lipid, anionic lipid, another ionizable lipid, or zwitterionic lipid.

[0172] In some embodiments, the other lipid component(s) in the lipid composition may comprise a sterol, a PEG-modified lipid, and / or a helper lipid. The lipid composition thus comprises the novel ionizable lipid described herein, a sterol, a PEG-modified lipid, and / or a helper lipid.

[0173] The role and identity of exemplary helper lipids for lipid nanoparticles are known to one skilled in the art and may be any lipid compound that contributes to the stability and delivery efficiency of the lipid composition (e.g., LNP), or to the stable encapsulation of a therapeutic agent within the lipid composition (e.g., LNP).

[0174] In certain embodiments, the helper lipid is a negatively charged (anionic) lipid.Examples of suitable anionic lipids include, but are not limited to dimyrystoyl-, dipalmitoyl-, and distearoyl-phasphatidylglycerol; dimyrystoyl-, dipalmitoyl-, and dipalmitoyl-phosphatidic acid; dimyrystoyl-, dipalmitoyl-, and dipalmitoyl-phosphatidylethanolamine; and their unsaturated diacyl and mixed acyl chain counterparts as well as cardiolipin.

[0175] In certain embodiments, the helper lipid is a positively charged (cationic) lipid.Examples of cationic lipids include, but are not limited to, N, N'-dimethyl-N, N'-dioctacyl ammonium bromide (DDAB, i.e., didodecyldimethylammonium bromide) and chloride DDAC), N-(l-(2, 3 -dioleyl oxy )propyl)-N, N, N-trimethylammonium chloride (DOTMA, i.e., 1,2-di-O-octadecenyl-3 -trimethylammonium propane), N-methyldioctadecylamine (MDOA), 3 -[N-(N', N'-dimethylaminoethyl)carbamoyl) cholesterol (DC-chol), l,2-dioleoyloxy-3-[trimethylammonio]-propane (DOTAP), l,2-dioctadecyloxy-3-[trimethylammonio]-propane (DSTAP), and l,2-dioleoyloxypropyl-3-dimethyl-hydroxy ethyl ammonium chloride (DORI).

[0176] In certain embodiments, the helper lipid is a neutral charged lipid. Examples of suitable- 63 - LEGAL\113833648\6neutral lipids include, but are not limited to DLPC (1,2-dilauroyl- sn-glycero-3 -phosphocholine), DMPC (l,2-dimyristoyl-sn-glycero-3 -phosphocholine), DPPC (l,2-dipalmitoyl-sn-glycero-3-phosphocholine), DSPC (l,2-distearoyl-sn-glycero-3- phosphocholine), DOPC (1,2-dioleoyl-sn-glycero-3 -phosphocholine), DMPA (sodium 1,2- di tetradecanoyl-sn-glycero-3 -phosphate), DPPE (l,2-dipalmitoyl-sn-glycero-3- phosphoethanolamine), and DOPE (l,2-dioleoyl-sn-glycero-3-phosphoethanolamine).

[0177] In some embodiments, the helper lipid is a phospholipid. Phospholipids may assemble into one or more lipid bilayers. In general, phospholipids comprise a phospholipid moiety and one or more fatty acid moieties. A phospholipid moiety can be selected, for example, from the non-limiting group consisting of phosphatidyl choline, phosphatidyl ethanolamine, phosphatidyl glycerol, phosphatidyl serine, phosphatidic acid, 2-lysophosphatidyl choline, and a sphingomyelin. A fatty acid moiety can be selected, for example, from the non-limiting group consisting of lauric acid, myristic acid, myristoleic acid, palmitic acid, palmitoleic acid, stearic acid, oleic acid, linoleic acid, alpha-linolenic acid, erucic acid, phytanoic acid, arachidic acid, arachidonic acid, eicosapentaenoic acid, behenic acid, docosapentaenoic acid, and docosahexaenoic acid.

[0178] Non-natural phospholipid species including natural species with modifications and substitutions including branching, oxidation, cyclization, and alkynes are also contemplated. In some embodiments, a phospholipid can be functionalized with or cross-linked to one or more alkynes (e.g., an alkenyl group in which one or more double bonds is replaced with a triple bond). Under appropriate reaction conditions, an alkyne group can undergo a copper-catalyzed cycloaddition upon exposure to an azide. Such reactions can be useful in functionalizing a lipid bilayer of a nanoparticle composition to facilitate membrane permeation or cellular recognition or in conjugating a nanoparticle composition to a useful component such as a targeting or imaging moiety (e.g., a dye).

[0179] Suitable phospholipids include, but are not limited to, glycerophospholipids such as phosphatidylcholines, phosphatidylethanolamines, phosphatidylserines, phosphatidylinositols, phosphatidy glycerols, and phosphatidic acids. Suitable phospholipids also include phosphosphingolipid, such as sphingomyelin.

[0180] Exemplary phospholipids are lecithin, phosphatidylethanolamine, lysolecithin, lysophosphatidylethanolamine(LPE), phosphatidylserine, phosphatidylinositol, sphingomyelin, egg sphingomyelin (ESM), phosphatidylethanolamine (cephalin), cardiolipin, phosphatidic acid, phosphatidylcholine, cerebrosides, dicetylphosphate, l,2-dioleoyl-sn-glycero-3 -phosphate (DOPA; 18:1 PA), 1, 2-distearoyl-sn-glycero-3 -phosphate (DSPA; 18:0 PA), distearoylphosphatidylcholine (DSPC), l,2-dilinoleoyl-sn-glycero-3 -phosphocholine (DLPC),- 64 - LEGAL\113833648\61.2-dimyristoyl-sn-glycero-phosphocholine (DMPC), dioleoylphosphatidylcholine (DOPC), dipalmitoylphosphatidylcholine (DPPC), dioleoylphosphatidylglycerol (DOPG), dipalmitoylphosphatidylglycerol (DPPG), dioleoylphosphatidyl ethanolamine (DOPE), 1,2-diundecanoyl-sn-glycero-phosphocholine (DUPC), palmitoyloleoyl-phosphatidylcholine (POPC), palmitoyloleoyl-phosphatidylethanolamine (POPE), palmitoyloleyol-phosphatidylglycerol (POPG), dioleoylphosphatidylethanolamine 4-(N-maleimidomethyl)-cyclohexane- 1 -carboxylate (DOPE-mal), dipalmitoyl-phosphatidylethanolamine (DPPE), dimyristoylphosphatidylethanolamine (DMPE), distearoyl-phosphatidylethanolamine (DSPE), monomethylphosphatidylethanolamine, dimethyl-phosphatidylethanolamine, dielaidoylphosphatidylethanolamine (DEPE), stearoyloleoyl -phosphatidylethanolamine (SOPE), 1 -stearoyl -2-oleoyl- phosphatidylcholine (SOPC), lysophosphatidylcholine, dilinoleoylphosphatidylcholine, 1.2-di-O-octadecenyl-sn-glycero-3 -phosphocholine (18:0 diether PC), l-oleoyl-2-cholesterylhemisuccinoyl-sn-glycero-3 -phosphocholine (OChemsPC), l-hexadecyl-sn-glycero-3-phosphocholine (C16Lyso PC), l,2-dilinolenoyl-sn-glycero-3 -phosphocholine, 1,2-diarachidonoyl-sn-glycero-3 -phosphocholine, l,2-didocosahexaenoyl-sn-glycero-3-phosphocholine, 1,2-diphytanoyl-sn-glycero-3 -phosphoethanolamine (ME 16.0 PE), 1,2-distearoyl-sn-glycero-3 -phosphoethanolamine, l,2-dilinoleoyl-sn-glycero-3-phosphoethanolamine, l,2-dilinolenoyl-sn-glycero-3 -phosphoethanolamine, 1,2-diarachidonoyl-sn-glycero-3 -phosphoethanolamine, l,2-didocosahexaenoyl-sn-glycero-3 -phosphoethanolamine, N, N-dicotadecylaniline, sn-(3-myristoyl-2-hydroxy)-glycerol-l-phospho-sn-3'-(l',2'-dimyristoyl)-glycerol (14:0 Hemi BMP), sphingomyelin, and mixtures thereof. Other diacylphosphatidylcholine and diacylphosphatidylethanolamine phospholipids can also be used. The acyl groups in these lipids may be acyl groups derived from fatty acids having C10-C24 carbon chains, e.g., lauroyl, myristoyl, palmitoyl, stearoyl, or oleoyl.

[0181] In some embodiments, the helper lipid is selected from the group consisting of DOPE, DOPC, DSPC, DSPE, DDAB, DOTMA, DOTAP, DOPA (18: 1 PA), MDOA, 14:0 Hemi BMP, DMPA (14:0 PA), DSPA (18:0 PA), and a combination thereof.

[0182] In some embodiments, the helper lipid is selected from the group consisting of DOPE, DDAB, DOTAP, DSPC, DSPE, DOTMA, and a combination thereof.

[0183] The lipid composition may also include a sterol or derivative thereof. In some embodiments, the sterol or derivative thereof is cholesterol, a cholesterol derivative, or a combination thereof. In some embodiments, a combination of two or more cholesterols or derivatives thereof are used. The inclusion of a cholesterol in nanoparticle formulations can improve efficacy, potentially due to enhanced membrane fusion. Exemplary cholesterol or derivatives thereof include cholesterol (C27H46O), 20a-OH cholesterol, 20a-hydroxycholesterol- 65 - LEGAL\113833648\6(5-cholestene-3p,20a-diol), and DC-cholesterol (N, N-dimethyl-N-ethylcarboxamidocholesterol). Any natural sterols may also be used for the cholesterol component. Examples of natural sterols include, for example, cholesterol sulfate, desmosterol, stigmasterol, lanosterol, 7-dehydrocholesterol, dihydrolanosterol, zymosterol, lathosterol, 14-demethyl-lanosterol, 8(9)-dehydrocholesterol, 8(14)-dehydrocholesterol, FF-MAS, diosgenin, DHEA sulfate, DHEA, sitosterol, lanosterol-95, cholesterol (plant), dihydro FF-MAS-d6, dihydro T-MAS-d6, zymostenol, sitostanol, campestanol, campesterol, 7-dehydrodesmosterol, pregnenolone, dihydro T-MAS, delta 5-avenasterol, brassicasterol, dihydro FF-MAS, and 24-methylene cholesterol. A large diversity of structural analogs of cholesterol exist as natural products (e.g., phytosterols that are plant-based sterols, which provide stability to the plant cell wall). Exemplary cholesterol analogs include, for example, Vitamin D derivatives (such as 9,10-secosteroids, Vitamin D2, Vitamin D3, Calcipotriol), alkyl-substituted steroids (such as C-24 alkyl steroids), and cholesterol analogs wherein the tail is modified into a fifth ring (such as pentacyclic steroids).

[0184] The lipid composition may also include a PEG-modified lipid. As used herein, the term “PEG-lipid” or “PEG-modified lipid” refers to polyethylene glycol (PEG)-modified lipids. Non-limiting examples of PEG lipids include PEG-modified phosphatidylethanolamine and phosphatidic acid, PEG-ceramide conjugates (e.g., PEG-CerC14 or PEG-CerC20), PEG-modified dialkylamines and PEG-modified 1,2-diacyloxypropan-3 -amines. Such lipids are also referred to as PEGylated lipids.

[0185] In some embodiment, the PEG-modified lipid is selected from the group consisting of a PEG-modified phosphatidylethanolamine, a PEG-modified phosphatidic acid, a PEG-modified ceramide, a PEG-modified dialkylamine, a PEG-modified diacylglycerol, a PEG-modified dialkylglycerol, and mixtures thereof.

[0186] In some embodiments, the lipid moiety of the PEG-modified lipid includes those having lengths of from about C to about C22. In some embodiments, the lipid moiety of the PEG lipids includes those having lengths of from about Cuto about Cis.

[0187] In some embodiments, the PEG lipid includes, but are not limited to, 1,2-dimyristoyl-sn-glycerol methoxypolyethylene glycol (PEG-DMG), 1,2-distearoyl-sn-gly cero-3 -phosphoethanolamine-N-[amino(polyethylene glycol)] (PEG-DSPE), PEG-disteryl glycerol (PEG-DSG), PEG-dipalmetoleyl, PEG-dioleyl, PEG-distearyl, PEG-diacylglycamide (PEGDAG), PEG-dipalmitoyl phosphatidylethanolamine (PEG-DPPE), PEG-l,2-dimyristoyl-sn-gly cero-3 -phosphoethanolamine (PEG-DMPE), PEG- 1,2-dilauroyl-sn-gly cero-3 -phosphoethanolamine (PEG-DLPE), PEG-dipalmitoylphosphatidylcholine (PEG-DPPC), PEG-l,2-dimyristyloxlpropyl-3 -amine (PEG-c-DMA), (l,2-dimyristoyl-3-propanoxy)-carboxamide-propyl-methoxypolyoxyethylene (PEG-c-DOMG).- 66 - LEGAL\113833648\6

[0188] The PEG moiety of the PEG-modified lipids described herein may comprise an average molecular weight ranging from 550 Daltons to 20,000 Daltons. In some embodiments, the PEG moiety has an average molecular weight of from 750 Daltons to 5,000 Daltons (e.g., from 1,000 Daltons to 5,000 Daltons, from 1,500 Daltons to 3,000 Daltons, from 750 Daltons to 3,000 Daltons, from 750 Daltons to 2,000 Daltons). For instance, a PEG moiety, such as an mPEG-NEE, may have an average molecular weight of about 1000, 2000, 5000, 10,000, 15,000 or 20,000 daltons. In some embodiments, the PEG moiety has an average molecular weight of 2,000 Daltons or 750 Daltons. In one embodiment, the PEG-modified lipid has PEG2K.

[0189] In one embodiment, the lipid composition described herein can comprise a PEG-modified lipid which is a non-diffusible PEG. Non-limiting examples of non-diffusible PEGs include PEG-DSG and PEG-DSPE.

[0190] In some embodiments, the PEG-modified lipid is PEG-DMG, PEG-DSG, PEG-PE, or PEG-DSPE.

[0191] In some embodiments, the PEG-modified lipid is DMPE-PEG2K (14:0 PEG2K PE), DSPE-PEG2K (18:0PEG2K PE), DMG-PEG2K, 2- [(polyethylene glycol)-2000]-N, N-ditetradecyl acetamide (ALC-0159), DSG-PEG2K, or a combination thereof.

[0192] In some embodiments, the lipid composition (e.g., LNP) comprises the compound described herein (e.g., including any compound of formulas (GI), (GII), (GT), (GIT), (PE-I), and / or (PE -II) and any of the compounds belonging to any subgenus formulas or species under formula (GI), (GII), (GT), (GIT), (PE-I), or (PE-II)), a helper lipid (e.g., a phospholipid such as DOPE, DDAB, DOTAP, DSPC, or a mixture thereof), a sterol (e.g., cholesterol, 20a-OH cholesterol, DC-cholesterol, or a mixture thereof), and a PEG-modified lipid (e.g., 14:0 PEG2K PE such as DMPE-PEG2K, 18:0PEG2K PE such as DSPE-PEG2K, DMG-PEG2K, DSG-PEG 2K, or a mixture thereof).

[0193] Molar ratios of the lipid compound described herein, the helper lipid, the sterol, and the PEG-modified lipid can be varied as needed. For example, the lipid composition can include, by mol% or wt% of the total lipid components, about 20-70% of an ionizable lipid (including the lipid compound as described herein, e.g., any compound of formulas (GI), (GII), (GI'), (GIF), (PE-I), and / or (PE-II) and any of the compounds belonging to any subgenus formulas or species under formula (GI), (GII), (GI'), (GIF), (PE-I), or (PE-II)), about 0-50% of the helper lipid, about 0-70% of the sterol, and about 0.5-10% of the PEG-modified lipid.

[0194] In some embodiments, in the lipid composition, the ionizable lipid including the lipid compounds described herein is present in an amount from about 20 mol% to about 100 mol% (e.g., 20-90 mol%, 20-80 mol%, 20-70 mol%, 20-60 mol%, 25-100 mol%, 30-70 mol%, 30-60 mol%, 30-40 mol%, 40-50 mol%, or 50-90 mol%) of the total lipid components; the helper lipid- 67 - LEGAL\113833648\6is present in an amount from about 0 mol% to about 50 mol% (e.g., 0-40 mol%, 0-30 mol%, 5-50 mol%, 5-40 mol%, 5-30 mol%, 5-20 mol%, 5-10 mol%, 7-50 mol%, 7-40 mol%, 7-30 mol%, 7-20 mol%, or 7-10 mol%) of the total lipid components; the sterol is present in an amount from about 0 mol % to about 70 mol% (e.g., 0-60 mol%, 0-50 mol%, 0-40 mol%, 0-35 mol%, 5-70 mol%, 5-60 mol%, 5-50 mol%, 5-40 mol%, 5-35 mol%, 10-70 mol%, 10-60 mol%, 10-50 mol%, 10-40 mol%, 10-35 mol%, 15-70 mol%, 15-60 mol%, 15-50 mol%, 15-40 mol%, 15-35 mol%, 20-70 mol%, 20-60 mol%, 20-50 mol%, 20-40 mol%, or 20-35 mol%) of the total lipid components; and the PEG-modified lipid is present in an amount from about 0.5 mol% to about 20 mol% (e.g., 0.5-15 mol%, 0.5-10 mol%, 0.5-5 mol%, 0.5-3 mol%, 0.5-2 mol%, 0.5-1.5 mol%, 1-15 mol%, 1-10 mol%, 1-5 mol%, or 1-3 mol%) of the total lipid components; provided that the total mol% of the lipid components does not exceed 100%.

[0195] In some embodiments, the lipid composition comprises: about 20-60 mol% of the lipid compound (e.g., any compound of formulas (GI), (GII), (GI'), (GIF), (PE-I), and / or (PE-II) and any of the compounds belonging to any subgenus formulas or species under formula (GI), (GII), (GP), (GIF), (PE-I), or (PE-II)); about 7-50 mol% of the helper lipid (e.g., a phospholipid such as DOPE, DDAB, DOTAP, DSPC, or a mixture thereof); about 5-70 mol% of the sterol (e.g., cholesterol, 20a-OH cholesterol, DC-cholesterol, or a mixture thereof); and about 0.5-3 mol% of the PEG-modified lipid (e.g., 14:0PEG2KPE such as DMPE-PEG2K, 18:0PEG2KPE such as DSPE-PEG2K, DMG-PEG 2K, DSG-PEG 2K, or a mixture thereof). In some embodiments, the lipid composition comprises: about 30-50 mol% of the lipid compound (e.g., any compound of formulas (GI), (GII), (GI'), (GIF), (PE-I), and / or (PE-II) and any of the compounds belonging to any subgenus formulas or species under formula (GI), (GII), (GF), (GIF), (PE-I), or (PE-II)); about 10-30 mol% of the helper lipid (e.g., a phospholipid such as DOPE, DDAB, DOTAP, DSPC, or a mixture thereof); about 35-50 mol% of the sterol (e.g., cholesterol, 20a-OH cholesterol, DC-cholesterol, or a mixture thereof); and about 1-3 mol% of the PEG-modified lipid (e.g., 14:0PEG2KPE such as DMPE-PEG2K, 18:0PEG2KPE such as DSPE-PEG2K, DMG-PEG 2K, DSG-PEG 2K, or a mixture thereof).

[0196] In some embodiments, in the lipid composition, the molar ratio of the lipid compound (e.g., any compound of formulas (GI), (GII), (GI'), (GIF), (PE-I), and / or (PE-II) and any of the compounds belonging to any subgenus formulas or species under formula (GI), (GII), (GI'), (GIF), (PE-I), or (PE-II)): the helper lipid (e.g., a phospholipid such as DOPE, DDAB, DOTAP, DSPC, or a mixture thereof): the sterol (e.g., cholesterol, 20a-OH cholesterol, DC-cholesterol, or a mixture thereof): the PEG-modified lipid (e.g., 14:0PEG2K PE such as DMPE-PEG2K, 18:0 PEG2K PE such as DSPE-PEG2K, DMG-PEG 2K, DSG-PEG 2K, or a mixture thereof) may be about 20-60:7-50:5-70:0.5-3. In certain embodiments, the molar ratio of the lipid compound: the- 68 - LEGAL\113833648\6helper lipid: the sterol: the PEG-modified lipid is about 25-55:10-50:15-50:0.5-3. In certain embodiments, the molar ratio of the lipid compound: the helper lipid: the sterol: the PEG-modified lipid is about 30-55:10-20:30-50:1-3. In some embodiments, the molar ratio of the lipid compound: the helper lipid: the sterol: the PEG-modified lipid is about 32-50:10-18:32-48:1-2.5. In certain embodiments, the molar ratio of the lipid compound: the helper lipid: the sterol: the PEG-modified lipid is about 35-50: 10-16:35-48: 1-2.5. In some embodiments, the molar ratio of the lipid compound: the helper lipid: the sterol: the PEG-modified lipid is about 30-50:10-30:35-50: 1-3.

[0197] In some embodiments, the molar ratio of the lipid compound: the helper lipid: the sterol: the PEG-modified lipid is about 35:15:47.5:2.5, about 35:16:46.5:2.5, about 45:9:44:2, about 45:13:39.5:2.5, about 50:17.5:30:2.5, about 50:12.5:35:2.5, or about 50:10:38.5:1.5.

[0198] The molar concentration of the lipid compound in the lipid composition (e.g., LNP) may be about 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, 50, 51, 52, 53, 54, 55, 56, 57, 58, 59, or 60 percent of the total lipids in the lipid composition (e.g., LNP). The molar concentration of the helper lipid in the lipid composition (e.g., LNP) may be about 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, or 50 percent of the total lipids in the lipid composition (e.g., LNP). The molar concentration of the sterol in the lipid composition (e.g., LNP) may be about 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, 50, 51, 52, 53, 54, 55, 56, 57, 58, 59, 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, or 70 percent of the total lipids in the lipid composition (e.g., LNP). The molar concentration of the PEG-modified lipid may be about 0.5, 0.75, 1, 1.25, 1.5, 1.75, 2, 2.25, 2.5, 2.75, or 3 percent of the total lipids in the lipid composition (e.g., LNP).

[0199] The lipid composition (e.g., liposome or LNP) may have a particle size (e.g., an average particle diameter) of about 20-250, 20-225, 20-200, 30-250, 30-225, 20-300, 40-250, 40-225, 40-200, 45-250, 45-225, 45-200, 50-200, 50-180, or 75-170 nm, e.g., measured by dynamic light scattering (DLS). For example, the lipid composition (e.g., liposome or LNP) may have a particle size (e.g., an average particle diameter) of about 20, 30, 40, 50, 60, 70, 80, 90, 100, 110, 120, 130, 140, 150, 160, 170, 180, 190, 200, 210, 220, 230, 240, or 250 nm. In a given population of particles of the lipid composition, the population may include individual members of respectively different sizes. The particle size distribution of a given population of the particles of the lipid composition (e.g., liposome or LNP) may be characterized by a D90 of about 250, 240, 230, 220, 210, 200, 190, 180, 170, 160, 150, 140, 130, 120, 110, 100, 95, 90, 85, 80, 75, 70, 65, or 60 nm, and / or a D10 of about 20, 30, 40, 50, 60, 70, 80, 90, 100, 110, 120, 130, 140, 150,- 69 - LEGAL\113833648\6160, 170, 180, 190, or 200 nm.

[0200] The lipid composition (e.g., liposome or LNP) may, in some instances, be relatively homogenous. A polydispersity index (PDI) may be used to indicate the homogeneity of a lipid composition (e.g., liposome or LNP), e.g., the particle size distribution of the LNP. A small (e.g., less than 0.3) polydispersity index generally indicates a narrow particle size distribution. A lipid composition (e.g., liposome or LNP) may have a polydispersity index of about 0.3 or less. For instance, the lipid composition (e.g., liposome or LNP) may have a PDI from about 0 to about 0.3, such as 0.01, 0.02, 0.03, 0.04, 0.05, 0.06, 0.07, 0.08, 0.09, 0.10, 0.11, 0.12, 0.13, 0.14, 0.15, 0.16, 0.17, 0.18, 0.19, 0.20, 0.21, 0.22, 0.23, 0.24, 0.25, 0.26, 0.27, 0.28, 0.29, or 0.3. In some embodiments, the PDI of the lipid composition (e.g., LNP) may be from about 0.10 to about 0.30.

[0201] The zeta potential of a lipid composition (e.g., liposome or LNP) may be used to indicate the electrokinetic potential of the composition. In some embodiments, the zeta potential may describe the surface charge of a liposome or LNP. Lipid compositions (e.g., liposome or LNP) with relatively low charges, positive or negative, are generally desirable, as more highly charged species may interact undesirably with cells, tissues, and other elements in the body. In some embodiments, the zeta potential of a liposome or LNP may be from about -10 mV to about +20 mV, from about -10 mV to about +15 mV, from about -10 mV to about +10 mV, from about -10 mV to about +5 mV, from about -10 mV to about 0 mV, from about -10 mV to about -5 mV, from about -5 mV to about +20 mV, from about -5 mV to about +15 mV, from about -5 mV to about +10 mV, from about -5 mV to about +5 mV, from about -5 mV to about 0 mV, from about 0 mV to about +20 mV, from about 0 mV to about +15 mV, from about 0 mV to about +10 mV, from about 0 mV to about +5 mV, from about +5 mV to about +20 mV, from about +5 mV to about +15 mV, or from about +5 mV to about +10 mV.

[0202] The lipid composition may include one or more components in addition to those lipid components described above. For example, the lipid composition may include one or more small hydrophobic molecules such as a vitamin (e.g., vitamin A. or vitamin E). The lipid composition may also include one or more permeability enhancer molecules, carbohydrates, polymers, surface altering agents, or other components. Suitable permeability enhancer molecules, carbohydrates, polymers, and surface altering agents include those well known to one skilled in the art for use in preparation of liposomes or LNPs.

[0203] The lipid composition may optionally include one or more coatings. In some embodiments, the lipid composition (e.g., liposome or LNP) may be formulated in a capsule, film, or tablet having a coating. A capsule, film, or tablet including a composition described herein may have any useful size, tensile strength, hardness, or density.

[0204] Additional exemplary lipids, formulations, methods, and characterization of a lipid- 70 - LEGAL\113833648\6composition (e.g., liposome or LNP) may be found in WO 2020 / 061457 and WO 2021 / 113777, which are incorporated herein by reference in their entirety. In some embodiments, the lipid compositions (e.g., liposome or LNP) include those that are formulated specifically for the delivery of CRISPR-Cas systems, e.g., Cas9-gRNA RNP, gRNA, Cas9 mRNA, details of which are described in WO 2019067992 and WO 2019067910, which are incorporated by reference in their entirety. Additional specific lipid compositions (e.g., liposome or LNP) useful for delivery of nucleic acids are described in U. S. Patent Nos. 8,158,601 and 8,168,775, which are incorporated by reference in their entirety.

[0205] Suitable methods for preparing the lipid compositions (e.g., liposomes or LNPs) include those known in the art. For example, suitable methods can include employing mixing processes such as microfluidics, including herringbone micromixing, and T-junction mixing of two fluid streams, one of which contains an RNA, e.g., mRNA, typically in an aqueous solution, and the other of which has the various required lipid components, typically in ethanol. Lipid particles (e.g., lipid nanoparticles) can form spontaneously upon mixing of the nucleic acid, e.g., mRNA, and the lipid(s). Depending on the desired particle size distribution, the resultant nanoparticle mixture can be extruded through a membrane (e.g., 100 nm cut-off) using, for example, a thermobarrel extruder. In some embodiments, the extrusion step can be omitted. Ethanol removal and simultaneous buffer exchange can be accomplished by, for example, dialysis or tangential flow filtration.

[0206] Additional suitable methods for preparing the lipid compositions (e.g., liposomes or LNPs) may be found, e.g., in WO 2022 / 261101; WO 2019051289; US 2013 / 0037977; US 2010 / 0015218; US 2013 / 0156845; US 2013 / 0164400; US 2012 / 0225129; US 2010 / 0130588; US 2007 / 0042031; US 2004 / 0142025; Lichtenberg andBarenholz in Methods of Biochemical Analysis, 33:337-462 (1988); Szoka et al., Ann. Rev. Biophys. Bioeng. 9:467 (1980); U. S. Patent Nos. 4,235,871; 4,501,728; and 4,837,028; Liposomes, Marc J. Ostro, ed., Marcel Dekker, Inc., New York, 1983, Chapter 1; and Hope, et al., Chem. Phys. Lip. 40:89 (1986), all of which are incorporated herein by reference in their entirety. In some embodiments, the lipid compositions (e.g., liposomes or LNPs) can be prepared using a continuous mixing method, a direct dilution process, or an in-line dilution process. The processes and apparatuses for apparatuses for preparing lipid nanoparticles using direct dilution and in-line dilution processes are described, e.g., in US 2007 / 0042031. The processes and apparatuses for preparing the lipid compositions (e.g., liposomes or LNPs) using stepwise dilution processes are described in US 2004 / 0142025.

[0207] The method for preparing the lipid compositions (e.g., LNPs) employing the novel ionizable lipids are further exemplified in Example 2 below (see, e.g., Exemplary preparation of LNP without conjugation to a targeting moiety (Untargeted LNPs)).- 71 - LEGAL\113833648\6Targeting Moiety

[0208] The lipid composition (e.g., liposomes or LNPs) may further comprise a targeting moiety conjugated to one or more lipid components of the lipid composition.

[0209] In some embodiments, the targeting moiety is a peptide, polypeptide, protein; a carbohydrate or polysaccharide; or a small molecule targeting ligand.

[0210] In some embodiments, the targeting moiety is an antibody or fragment thereof, having a binding affinity to a cell. In some embodiments, the targeting moiety is a monoclonal antibody, single-chain variable fragment (scFv), cytokine, growth factor, or receptor-binding domain.

[0211] In some embodiments, the targeting moiety is an antibody or fragment thereof that binds CD markers involved in activation, maturation, and / or subset differentiation for T cells (e.g., CD3, CD5, CD7, CD8+, CD57, etc.). In some embodiments, the targeting moiety is an antibody or fragment thereof that binds hepatocyte-specific surface receptors and expression proteins (e.g, ASGPR, LDLR, TfR, Fc receptor, CD206, CD163, VEGF / VEGFR, etc ).

[0212] In some embodiments, the targeting moiety is covalently attached to the one or more lipid components of the lipid composition.

[0213] Conjugation of the targeting moiety (e.g., an antibody or fragment thereof) to the lipid composition may be carried out by functionalizing the targeting moiety (e.g., an antibody or fragment thereof) with a first functional group, and functionalizing a lipid component of the lipid composition with a second functional group, wherein the first and second functional groups are reactive or interactive to each other to form the conjugation.

[0214] For example, the conjugation of the targeting moiety (e.g., an antibody or fragment thereof) to a lipid component of the lipid composition may be carried out via one or more of the following reactions or interactions:a Sortase A-mediated site-specific functionalization of C-terminus of the antibody or fragment thereof with an azide group,a click reaction between an azide and an alkyne (e.g., a terminal alkyne or a cycloalkyne such as bicyclononyne (BCN) or dibenzocyclooctyne (DBCO)),a click reaction between an alkene (e.g., a trans cyclooctene) and a tetrazine (such as a tetrazine-trans-cyclooctene ligation),a click reaction between an alkyne (e.g., a terminal alkyne or a cycloalkyne such as BCN or DBCO) and a tetrazine,a thiol reaction between the thiol group of a cysteine and a thiol-reactive group (such as a thiol-maleimide reaction, a thiol-disulfide interchange reaction, a thiol-thiol ester reaction, a- 72 - LEGAL\113833648\6thiol-haloacetyl reaction, a thiol-alkene reaction, a thiol-alkyne reaction, a thiol-vinyl sulfone reaction, a thiol-acrylamide reaction, etc.),a NHS ester reaction between an NHS ester and a primary amine, andanon-covalent ligand-receptor coupling (e.g., biotin-streptavidin).

[0215] More details on various reactions and mechanisms for conjugation of a targeting moiety (e.g., an antibody or fragment thereof) to a lipid component of a lipid composition (e.g., LNP) may be found in e.g., WO 2023 / 248125A1, US 2025 / 0161481, and Chen et al., Journal of Controlled Release 388: 114365 (2025), all of which are incorporated herein by reference in their entirety.

[0216] In some embodiments, the lipid composition contains a lipid component that comprises one or more PEG-modified lipids, and functionalizing the lipid component is carried out by functionalizing the PEG-modified lipid.

[0217] In some embodiments, the lipid composition contains two or more PEG-modified lipids: at least one PEG-modified lipid is not functionalized and at least one PEG-modified lipid is functionalized. The PEG-modified lipid that is not functionalized can be any PEG-modified lipid discussed herein. The two or more PEG-modified lipids may be based on the same or different PEG-modified lipid. The functionalized PEG-modified lipid can be any PEG-modified lipid discussed herein with the functionalization discussed herein. For instance, a lipid composition may contain two PEG-modified lipids, one is DSG-PEG2000, and the other one is a functionalized DSPE-PEG2000 (e.g., DSPE-PEG2000-TCO, trans-cyclooctene) for conjugation to a targeting moiety that is functionalized with a TCO reactive group (e.g., a tetrazine functionalized antibody). As another example, a lipid composition may contain two PEG-modified lipids, one is DMG-PEG2000, and the other one is a functionalized DSPE-PEG2000 (e.g., DSPE-PEG2000-TCO) for conjugation to a targeting moiety that is functionalized with a TCO reactive group (e.g., a tetrazine functionalized antibody).

[0218] In some embodiments, in the PEG-modified lipid component, the molar ratio of the PEG-modified lipid that is not functionalized to the functionalized PEG-modified lipid is about 1:1 or greater, about 2:1 or greater (e.g., 1:0.5), about 5:1 or greater, about 10:1 or greater, about 15:1 or greater (e.g., 1.4:0.1), about 20:1 or greater, about 30:1 or greater (e.g., 1.45:0.05), about 40:1 or greater, about 50:1 or greater, about 60:1 or greater, about 70:1 or greater, about 80:1 or greater, about 90:1 or greater, about 100:1 or greater, about 150:1 or greater (e.g., 1.49:0.01), about 200: 1 or greater, or about 300 or greater. Non-limiting exemplary molar ratios of the PEG-modified lipid that is not functionalized to the functionalized PEG-modified lipid include 1.0:0.5, 1.4:0.1, 1.45:0.05, and 1.49:0.01.

[0219] In the embodiments where the lipid composition contains two or more PEG-modified- 73 - LEGAL\113833648\6lipids: at least one PEG-modified lipid is not functionalized and at least one PEG-modified lipid is functionalized, all PEG-modified lipids (functionalized or not functionalized) may be considered as constituting the lipid component of PEG-modified lipid. Thus, all above descriptions and all embodiments discussed in the above aspects relating to the lipid composition, including various lipid components including the PEG-modified lipid and the ratios of various lipid components in the lipid composition are applicable herein to the conjugated lipid composition. As an example, in an embodiment above describing the molar ratio of the lipid compound: the helper lipid: the sterol: the PEG-modified lipid being about 50:10:38.5:1.5, the same molar ratio for the four lipid components would still hold for the targeting-moiety conjugated lipid composition. When the targeting moiety is conjugated to the functionalized PEG-modified lipid, the total amount of functionalized PEG-modified lipid and the unfunctionalized PEG-modified lipid constitutes the 1.5 mol% for the PEG-modified lipid component. For instance, for the above lipid composition, when the molar ratio of the unfunctionalized PEG-modified lipid to functionalized PEG-modified lipid is 2: 1, the molar ratio of the lipid compound: the helper lipid: the sterol: the unfunctionalized PEG-modified lipid: functionalized PEG-modified lipid would be about 50:10:38.5:1.0:0.5.

[0220] The method for preparing the lipid compositions (e.g., LNPs) that are functionalized with a targeting moiety is further exemplified in Example 2 below (see, e.g., CD5-Targeting Ligand Preparation and Exemplary preparation of LNP conjugated to a targeting moiety (Targeted LNPs)).Therapeutic Agent

[0221] The lipid composition (e.g., liposome or LNP) described herein can be used for the delivery of a therapeutic agent to a living organism, such as to a human subject. The therapeutic agent may be loaded to the lipid composition, e.g., encapsulated within the lipid composition (e.g., liposome or LNP). In some embodiments, the lipid composition further comprises a therapeutic agent.

[0222] Thus, in some embodiments, provided herein is a therapeutic agent-loaded lipid composition, comprising:a therapeutic agent;one or more compounds of formula (GI), (GII), (GF), (GIF), (PE -I), or (PE -II); or any of the compounds belonging to any subgenus formulas or species under formula (GI), (GII), (GI'), (GIF), (PE -I), or ( PE - 11 ) disclosed herein;a helper lipid;a sterol; and- 74 - LEGAL\113833648\6a PEG-modified lipid.

[0223] In some embodiments, the therapeutic agent may be a nucleic acid molecule (e.g., an oligonucleotide), a protein or peptide, a carbohydrate or glycoprotein, lipid, a small molecule, or any combination thereof. The lipid composition can contain two or more different therapeutic agents from the nucleic acid molecule, peptide or protein, carbohydrate or glycoprotein, and small molecule drug.

[0224] In some embodiments, the therapeutic agent is a small molecule drug, for instance, a small molecule drug approved for use in humans by an appropriate regulatory authority. In some embodiments, the lipid composition can include a plurality of small molecule drugs, which may be the same or different types.

[0225] In some embodiments, the therapeutic agent is a carbohydrate or glycoprotein. In some embodiments, the therapeutic agent may be a simple sugar (e g., glucose) or a polysaccharide (e.g., glycogen and derivatives and analogs thereof). In some embodiments, the therapeutic agent is a glycoprotein. In some embodiments, the lipid composition may include a plurality of carbohydrates or glycoproteins, which may be the same or different types.

[0226] In some embodiments, the therapeutic agent is a protein or peptide. In some embodiments, the protein may be a peptide or polypeptide, e.g., a transcription factor; a chromatin remodeling factor; an antigen; a hormone; an enzyme (such as a nuclease, e.g., an endonuclease, e.g., a nuclease element of a CRISPR system, e.g., a Cas9, dCas9, aCas9-nickase, Cpf / Casl2a); a Crispr-linked enzyme, e.g., a base editor or prime editor; a mobile genetic element protein (e.g., a transposase, a retrotransposase, a recombinase, an integrase); a gene writer; a polymerase; a methylase; a demethylase; an acetylase; a deacetylase; a kinase; a phosphatase; a ligase; a deubiquitinase; an integrase; a recombinase; a topoisomerase; a gyrase; a helicase; a lysosomal acid hydrolase); an antibody; a receptor ligand; a receptor; a clotting factor; a membrane protein; a mitochondrial protein; a nuclear protein; an antibody or other protein scaffold binder such as a centyrin, darpin, or adnectin. In some embodiments, the lipid composition may include a plurality of proteins or peptides, which may be the same or different types.

[0227] In some embodiment, the therapeutic agent is a nucleic acid molecule (e.g., DNA or RNA). In some embodiments, the nucleic acid molecule is a plasmid, an immunostimulatory oligonucleotide, an antisense oligonucleotide, an antagomir, an aptamer, a deoxyribozyme (DNAzyme), or a ribozyme.

[0228] In some embodiments, the therapeutic agent is a DNA, e.g., any DNA known to one skilled in the art. In some embodiments, the DNA is antisense DNA, plasmid DNA, parts of a plasmid DNA, pre-condensed DNA, a product of a polymerase chain reaction (PCR), vectors- 75 - LEGAL\113833648\6(e.g., PI, PAC, BAC, YAC, artificial chromosomes), expression cassettes, chimeric sequences, chromosomal DNA, or derivatives of these groups. In some embodiments, the DNA is linear DNA, circular DNA, single stranded DNA, or double stranded DNA.

[0229] In some embodiments, the therapeutic agent is an RNA, e.g., any RNA known to one skilled in the art. In some embodiments, the DNA is messenger RNA (mRNA), ribosomal RNA (rRNA), signal recognition particle RNA (7 SL RNA or SRP RNA), transfer RNA (tRNA), transfer-messenger RNA (tmRNA), small nuclear RNA (snRNA), small nucleolar RNA (snoRNA), SmY RNA, small Cajal body-specific RNA (scaRNA), guide RNA (gRNA), ribonuclease P (RNase P), Y RNA, telomerase RNA component (TERC), spliced leader RNA (SL RNA), antisense RNA (aRNA or asRNA), cis-natural antisense transcript (cis-NAT), CRISPR RNA (crRNA), long noncoding RNA (IncRNA), microRNA (miRNA), piwi-interacting RNA (piRNA), small interfering RNA (siRNA), transacting siRNA (tasiRNA), repeat associated siRNA (rasiRNA), 73K RNA, retrotransposons, a viral genome, a viroid, satellite RNA, or derivatives of these groups.

[0230] In some embodiments, the therapeutic agent is an mRNA.

[0231] In some embodiments, the therapeutic agent is an enzymatic nucleic acid molecule. The term “enzymatic nucleic acid molecule” refers to a nucleic acid molecule which has complementarity in a substrate binding region to a specified gene target, and also has an enzymatic activity which is active to specifically cleave target RNA. The term “enzymatic nucleic acid” is used interchangeably with phrases such as ribozymes, catalytic RNA, enzymatic RNA, catalytic DNA, aptazyme or aptamer-binding ribozyme, regulatable ribozyme, catalytic oligonucleotides, nucleozyme, DNAzyme, RNA enzyme, endoribonuclease, endonuclease, minizyme, leadzyme, oligozyme or DNA enzyme. All of these terminologies describe nucleic acid molecules with enzymatic activity.

[0232] In some embodiments, the therapeutic agent is an antisense nucleic acid. The term “antisense nucleic acid” refers to a non-enzymatic nucleic acid molecule that binds to target RNA by means of RNA-RNA or RNA-DNA or RNA-PNA (protein nucleic acid) interactions and alters the activity of the target RNA.

[0233] In some embodiments, the nucleic acid molecule may be a 2-5A antisense chimera. The term “2-5 A antisense chimera” refers to an antisense oligonucleotide containing a 5'-phosphorylated 2'-5 '-linked adenylate residue.

[0234] In some embodiments, the nucleic acid molecule may be a triplex forming oligonucleotide. The term “triplex forming oligonucleotide” refers to an oligonucleotide that can bind to a double-stranded DNA in a sequence-specific manner to form a triple-strand helix.

[0235] In some embodiments, the nucleic acid molecule may be a decoy RNA. The term- 76 - LEGAL\113833648\6“decoy RNA” refers to a RNA molecule or aptamer that is designed to preferentially bind to a predetermined ligand. Such binding can result in the inhibition or activation of a target molecule.

[0236] In some embodiments, the nucleic acid molecule (e.g., RNA or DNA) encodes a therapeutic peptide or polypeptide, operably linked to a promoter for a DNA. The therapeutic peptide or polypeptide may be, e.g., a transcription factor; a chromatin remodeling factor; an antigen; a hormone; an enzyme (such as a nuclease, e.g., an endonuclease, e.g., a nuclease element of a CRISPR system, e.g., a Cas9, dCas9, aCas9-nickase, Cpf / Casl2a); a Crispr-linked enzyme, e.g., a base editor or prime editor; a mobile genetic element protein (e.g., a transposase, a retrotransposase, a recombinase, an integrase); a gene writer; a polymerase; a methylase; a demethylase; an acetylase; a deacetylase; a kinase; a phosphatase; a ligase; a deubiquitinase; an integrase; a recombinase; a topoisomerase; a gyrase; a helicase; a lysosomal acid hydrolase); an antibody; a receptor ligand; a receptor; a clotting factor; a membrane protein; a mitochondrial protein; a nuclear protein; an antibody or other protein scaffold binder such as a centyrin, darpin, or adnectin.

[0237] In some embodiments, the nucleic acid molecule comprises a Class 2 Cas nuclease mRNA and a gRNA. In some embodiments, the gRNA nucleic acid is or encodes a dual-guide RNA (dgRNA). In some embodiments, the gRNA nucleic acid is or encodes a single-guide RNA (sgRNA). In some embodiments, the gRNA is a modified gRNA. In some embodiments, the modified gRNA comprises a modification at one or more of the first five nucleotides at a 5’ end. In some embodiments, the modified gRNA comprises a modification at one or more of the last five nucleotides at a 3’ end.

[0238] In some embodiments, the nucleic acid molecule comprises an RNA-guided DNA-binding agent, for example a Cas nuclease mRNA (such as a Class 2 Cas nuclease mRNA) or a Cas9 nuclease mRNA.

[0239] In some embodiments, the nucleic acid molecule can include a plurality of sequences. The plurality may be the same or different types. The plurality of sequences may be the same or different sequences of the same type.

[0240] All the nucleic acid molecules described herein can be chemically modified. The various modification strategy to the nucleic acid molecules are well known to one skilled in the art. In some embodiments, the nucleic acid molecule comprises one or more modifications selected from the group consisting of pseudouridine, 5 -bromouracil, 5-methylcytosine, peptide nucleic acid, xeno nucleic acid, morpholinos, locked nucleic acids, glycol nucleic acids, threose nucleic acids, dideoxynucleotides, cordycepin, 7-deaza-GTP, florophores (e.g. rhodamine or fluorescein linked to the sugar), thiol containing nucleotides, biotin linked nucleotides, fluorescent base analogs, CpG islands, methyl-7-guanosine, methylated nucleotides, inosine,- 77 - LEGAL\113833648\6thiouridine, pseudourdine, dihydrouridine, queuosine, and wyosine. In some embodiments, the antisense oligonucleotide may be a locked nucleic acid oligonucleotide (LNA). The term “locked nucleic acid (LNA)” refers to oligonucleotides that contain one or more nucleotide building blocks in which an extra methylene bridge fixes the ribose moiety either in the C3'-endo (beta-D-LNA) or C2'-endo (alpha-L-LNA) conformation (Grunweller A, Hartmann R K, BioDrugs, 21(4): 235-243 (2007), which is herein incorporated by reference in its entirety).

[0241] Additional examples of the nucleic acid molecules (including tumor suppressor genes, antisense oligonucleotides, siRNA, miRNA, or shRNA) may be found in US 2007 / 0065499 and U. S. Patent No. 7,780,882, which are incorporated by reference herein in their entireties.

[0242] In the lipid composition containing the therapeutic agent, the ratio of total lipid components to the therapeutic agent (e.g., an encapsulated therapeutic agent such as a nucleic acid molecule) can be varied as desired. For example, the total lipid components to the therapeutic agent (mass or weight) ratio can be from about 10: 1 to about 30: 1. In some embodiments, the total lipid components to the therapeutic agent ratio (mass / mass ratio; w / w ratio) can be in the range of from about 1: 1 to about 25: 1, from about 10:1 to about 14: 1, from about 3:1 to about 15:1, from about 4:1 to about 10:1, from about 5:1 to about 9:1, or about 6:1 to about 9:1. The amounts of total lipid components and the therapeutic agent can be adjusted to provide a desired N / P ratio. Generally, the lipid composition’s overall lipid content can range from about 5 mg / ml to about 30 mg / mL.

[0243] In some embodiments, the N / P ratio of the nucleic acid molecule-encapsulated lipid composition can range from about 1:1 to 30:1, for instance, from 3:1 to 20:1, from 3:1 to 15:1, from 3: 1 to 10: 1, or from 3: 1 to 6: 1. An N / P ratio refers to the molar ratio of the amines present in the lipid composition (e.g., the amines in the ionizable lipid compound) to the phosphates present in the nucleic acid molecule. It is a factor for efficient packaging and potency. In one embodiment, the N / P ratio of the nucleic acid molecule - encapsulated lipid composition ranges from 3: 1 to 15: 1.

[0244] The efficiency of encapsulation of the therapeutic agent (e.g., a protein or nucleic acid molecule) describes the amount of the therapeutic agent that is encapsulated or otherwise associated with the lipid composition (e.g., liposome or LNP) after preparation, relative to the initial amount provided. The encapsulation efficiency is desirably high (e.g., at least 70%. 80%.90%. 95%, close to 100%). The encapsulation efficiency may be measured, for example, by comparing the amount of the therapeutic agent in a solution containing the liposome or LNP before and after breaking up the liposome or LNP with one or more organic solvents or detergents. An anion exchange resin may be used to measure the amount of free protein or nucleic acid (e.g., RNA) in a solution. Fluorescence may be used to measure the amount of free- 78 - LEGAL\113833648\6protein and / or nucleic acid (e.g., RNA) in a solution. For the liposome or LNP described herein, the encapsulation efficiency of a therapeutic agent may be at least 50%, for example 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100%. In some embodiments, the encapsulation efficiency may be at least 80%. In some embodiments, the encapsulation efficiency may be at least 90%. In some embodiments, the encapsulation efficiency may be at least 95%.Pharmaceutical Composition and Formulations

[0245] Aspects of the invention also relate to a pharmaceutical composition comprising the lipid composition (e.g., liposome or LNP), which contains the lipid compound described herein (including one or more compounds of formula (GI), (GII), (GF), (GIF), (PE -I), or (PE -II); or any of the compounds belonging to any subgenus formulas or species under formula (GI), (GII), (GI'), (GIF), (PE -I), or (PE-II) disclosed herein) and which is loaded with the therapeutic agent described herein, and a pharmaceutically acceptable carrier.

[0246] In some embodiments, the pharmaceutical composition comprises a pharmaceutically acceptable excipient, and a lipid composition comprising:a therapeutic agent;one or more compounds of formula (GI), (GII), (GI'), (GIF), (PE -I), or (PE-II); or any of the compounds belonging to any subgenus formulas or species under formula (GI), (GII), (GI'), (GIF), (PE -I), or (PE-II) disclosed herein;a helper lipid;a sterol; anda PEG-modified lipid.

[0247] All above descriptions and all embodiments discussed in the above aspects relating to the novel lipid compounds, including the various genus and subgenus structural formulas, variable definitions and exemplary variables, and exemplary compounds, are all applicable to this aspect of the invention relating to the pharmaceutical composition.

[0248] All above descriptions and all embodiments discussed in the above aspects relating to the lipid composition, including various other lipid components and the therapeutic agents, are applicable to these aspects of the invention relating to the pharmaceutical composition.

[0249] The pharmaceutical compositions may contain one or more pharmaceutically acceptable excipients. The pharmaceutically acceptable excipient is selected on the basis of the mode and route of administration. Suitable pharmaceutical carriers or excipients for use in pharmaceutical formulations are described in Remington: The Science and Practice of Pharmacy, 21stEd., Gennaro, Ed., Lippencott Williams & Wilkins (2005); Handbook of Pharmaceutical- 79 - LEGAL\113833648\6Excipients, 6thEdition, Rowe et al., Eds., Pharmaceutical Press (2009); and the USP / NF (United States Pharmacopeia and the National Formulary), which are herein incorporated by reference in their entirety.

[0250] In some embodiments, the pharmaceutically acceptable excipient includes one or more of an antioxidant, binder, antiadherent, buffer, coloring agent, diluent (e.g., solid or liquid), disintegrant (e.g., coatings disintegrate), dispersing agent, dyestuff, filler, emulsifier, flavoring agent, lubricant, pH adjuster, pigment, preservative, stabilizer, solubilizing agent, solvent, suspending agent, sweetener, or wetting agent, or combination thereof.

[0251] Examples of suitable excipients include acacia, alginate, calcium phosphate, calcium carbonate, calcium silicate, carbopol gel, carboxymethyl cellulose, carnauba wax, cellulose, crospovidone, dextrose, diacetylated monoglycerides, ethylcellulose, gelatin, glyceryl monostearate 40-50, gum acacia, gum arabic, hydroxyethyl cellulose, hydroxypropyl cellulose, hydroxypropylmethyl cellulose, hypromellose phthalate, hypromellose, lactose, lecithin, magnesium stearate, kaolin, methacrylic acid copolymer type C, mannitol, methyl cellulose, methylhydroxybenzoate, microcrystalline cellulose, povidone, polyethylene glycol, polysorbate 80, polyvinylpyrrolidone, propylhydroxybenzoate, sodium carboxymethyl cellulose sodium hydroxide, sodium stearyl fumarate, sodium starch glycolate, starch, sorbitan monooleate sorbitol, sorbic acid, sucrose, talc, tragacanth, talc, triethyl citrate, titanium dioxide, yellow ferric oxide, talc, oil medium (e.g., peanut oil, liquid paraffin, mineral oil, olive oil, almond oil, glycerin, propylene glycol), or water,

[0252] When the excipient serves as a diluent, it can be a solid, semisolid, or liquid material (e.g., normal saline), which acts as a vehicle, carrier or medium for the active ingredient. As is known in the art, the type of diluent can vary depending upon the intended route of administration.

[0253] The pharmaceutical compositions can comprise pharmaceutically acceptable carriers, excipients, or stabilizers in the form of lyophilized formulations or aqueous solutions. Acceptable carriers, excipients, or stabilizers are nontoxic to recipients at the dosages and concentrations used, and may comprise buffers such as phosphate, citrate, and other organic acids; antioxidants including ascorbic acid and methionine; preservatives (such as octadecyldimethylbenzyl chloride; hexamethonium chloride, benzalkonium chloride, benzethonium chloride; phenol, butyl or benzyl alcohol; alkyl parabens such as methyl or propyl paraben; catechol; resorcinol; cyclohexanol; 3- pentanol; and m-cresol); low molecular weight (less than about 10 residues) polypeptides; proteins, such as serum albumin, gelatin, or immunoglobulins; hydrophilic polymers such as polyvinylpyrrolidone; amino acids such as glycine, glutamine, asparagine, histidine, arginine, or lysine; monosaccharides, disaccharides, and other carbohydrates including glucose, mannose, or- 80 - LEGAL\113833648\6dextrans; chelating agents such as EDTA; sugars such as sucrose, mannitol, trehalose or sorbitol; salt- forming counter-ions such as sodium; metal complexes (e.g., Zn- protein complexes); and / or non-ionic surfactants such as TWEEN™, PLURONICS™ or polyethylene glycol (PEG).

[0254] Suitable earners or excipients for the pharmaceutical compositions may also include a substance that enhances the ability of the body of an individual to absorb the LNP or liposome. Suitable carriers and / or excipients also include any substance that can be used to bulk up formulations with a LNP or liposome, to allow for convenient and accurate dosage. In addition, carriers and / or excipients may be used in the manufacturing process to aid in the handling of a LNP or liposome. Depending on the route of administration, and form of medication, different carriers and / or excipients may be used.

[0255] C arners and / or excipients may also include vehicles and / or diluents. “ Vehicles” indicates any of various media acting usually as solvents or carriers; “diluent” indicates a diluting agent which is issued to dilute an active ingredient of a composition; suitable diluent include any substance that can decrease the viscosity of a medicine. The type and amounts of carriers and / or excipi en ts are chosen in function of the chosen pharmaceutical form; suitable pharmaceutical forms are liquid systems like solutions, infusions, suspensions; semisolid systems like colloids, gels, pastes or creams; solid systems like powders, granulates, tablets, capsules, pellets, microgranulates, mini tablets, microcapsules, micropellets, suppositories, etc.

[0256] Each of the above systems can be suitably formulated for normal, delayed or accelerated release, using techniques well-known in the art.Formulations and Routes of administration

[0257] The lipid composition or the pharmaceutical composition containing the lipid composition described herein may be formulated according to standard techniques for any type of administration. For instance, the pharmaceutical compositions can be manufactured in a conventional manner, e.g., by conventional mixing, dissolving, granulating, dragee-making, levigating, emulsifying, encapsulating, entrapping, or lyophilizing processes. Methods well known in the art for making formulations are known in the art. See, e.g., Remington: The Science and Practice of Pharmacy, 21stEd., Gennaro, Ed., Lippencott Williams & Wilkins (2005), and Encyclopedia of Pharmaceutical Technology, eds. J. Swarbrick and J. C. Boylan, 1988-1999, Marcel Dekker, New York, which is incorporated herein by reference in its entirety.

[0258] For example, the pharmaceutical compositions may be formulated for administration orally, topically, parenterally, enterally, or by inhalation (e.g., intranasally). The active agent may be formulated for neat administration, or in combination with conventional pharmaceutical carriers, diluents, or excipients, which may be liquid or solid. The applicable solid carrier,- 81 - LEGAL\113833648\6diluent, or excipient may function as, among other things, a binder, disintegrant, filler, lubricant, glidant, compression aid, processing aid, color, sweetener, preservative, suspensing / dispersing agent, tablet-disintegrating agent, encapsulating material, film former or coating, flavoring agent, or printing ink. Any material used in preparing any dosage unit form is preferably pharmaceutically pure and substantially non-toxic in the amounts employed. In addition, the lipid composition (e.g., liposome or LNP) may be incorporated into sustained-release preparations and formulations.

[0259] In some embodiments, the pharmaceutical compositions may be formulated for parenteral administration, including intracanalicular administration, intravenous administration, subcutaneous administration, or intramuscular administration. As used herein, the term “parenteral” refers to routes of administration aside from enteral administration. Examples of parenteral administration include, e.g., buccal, epicutaneous, epidural, extra-amniotic, intraarterial, intra-articular, intracardiac, intracavemous, intracerebral, intracerebroventricular, intradermal, intralesional, intramuscular, intraocular, intraosseous infusion, intraperitoneal, intrapulmonary, intrathecal, intrauterine, intravaginal, intravenous, intravesical, intravitreal, nasal, perivascular, subcutaneous, sublingual, transdermal, topical, transepithelial, or transmucosal. Parenteral administration may be by continuous infusion over a selected period of time.

[0260] In some embodiments, the pharmaceutical compositions are administered intravenously by a bolus injection or infusion.

[0261] In some embodiments, the pharmaceutical composition is formulated for injection, such as intravenous infusion. The pharmaceutical forms suitable for injectable use include, for example, sterile aqueous solutions or dispersions and sterile powders for the extemporaneous preparation of sterile injectable solutions or dispersions. In all cases, the form is preferably sterile and fluid to provide easy syringability. It is preferably stable under the conditions of manufacture and storage and is preferably preserved against the contaminating action of microorganisms such as bacteria and fungi. The carrier, diluent, or excipient may be a solvent or dispersion medium containing, for example, water, ethanol, polyol (for example, glycerol, propylene glycol, liquid polyethylene glycol and the like), suitable mixtures thereof, and vegetable oils. The proper fluidity can be maintained, for example, by the use of a coating, such as lecithin, by the maintenance of the required particle size in the case of a dispersion, and by the use of surfactants. The prevention of the action of microorganisms may be achieved by various antibacterial and antifungal agents, for example, parabens, chlorobutanol, phenol, sorbic acid, thimerosal, and the like. In some embodiments, isotonic agents, for example, sugars or sodium chloride, may be included. Prolonged absorption of the injectable compositions may be achieved by the use of agents delaying absorption, for example, aluminum monostearate and gelatin.- 82 - LEGAL\113833648\6

[0262] Sterile injectable solutions may be prepared by incorporating the lipid composition in the pharmaceutically appropriate amounts, in the appropriate solvent, with various of the other ingredients enumerated above, as required, followed by filtered sterilization. Generally, dispersions may be prepared by incorporating the lipid composition into a sterile vehicle which contains the basic dispersion medium and the required other ingredients from those enumerated above. In the case of sterile powders for the preparation of sterile injectable solutions, the preferred methods of preparation may include vacuum drying and freeze drying techniques that yield a powder of the lipid composition or ingredients, plus any additional desired ingredient from the previously sterile-filtered solution thereof.

[0263] Any of the pharmaceutical compositions described herein can be used for delivering a therapeutic agent (such as a nucleic acid molecule) encapsulated in the lipid composition to a desired target To practice this use, an effective amount of a pharmaceutical composition as described herein can be administered to a subject in need of the treatment (e.g., a human subject) via a suitable route, such as those described herein.

[0264] The disclosure also provides dosage units containing the lipid composition or pharmaceutical composition disclosed herein. One skilled in the art would be able to select a dosage form for use herein. For example, the dosage unit may be a solid dosage form, liquid dosage form, or solid / liquid dosage form. In some embodiments, the dosage unit is a solid dosage form. In some embodiments, the dosage form is a liquid dosage form.

[0265] The dosage unit may be formulated for the delivery that is most useful to the subject. In some embodiments, the dosage unit is for enteral or parenteral administration. Examples of enteral administration include, but are not limited to, oral, rectal, sublingual, or buccal.

[0266] In some embodiments, the dosage unit is administered intravenously, intraperitoneally, intramuscularly, or subcutaneously. In some embodiments, the dosage unit is administered orally, intravenously, intraperitoneally, intramuscularly, or subcutaneously. In some embodiments, the dosage unit is administered orally.

[0267] In some embodiments, the dosage unit is for parenteral administration, i.e., a parenteral dosage unit. Parenteral dosage units are known in the art and include, but are not limited to, injectable solutions, inhalants, infusions, patches, and suppositories. In some embodiments, the parenteral dosage unit is an injectable solution. In some embodiments, the dosage unit is formulated for oral delivery, i.e., an oral dosage unit. In certain aspects, the oral dosage unit is a pill (e.g., tablet, caplet, capsule (e.g., soft gelatin, hard gelatin, gel capsule)), effervescent dosage form, elixir, fdm, liquid / solution (e.g., suspension, emulsion), lollipop, lozenge, paste, powder, sachet, or syrup. In some embodiments, the oral dosage unit is a pill, tablet, capsule, syrup, liquid solution, powder, paste, patch, pump, or fdm. In some embodiments, the oral dosage unit is a dry- 83 - LEGAL\113833648\6product for reconstitution with water or other suitable vehicle before use.

[0268] When the dosage form is a solid dosage form, an enteric coating can be applied or the solid dosage form may be scored. An enteric coating can be stable at low pH (e.g., in the stomach) and can dissolve at higher pH (e.g., in the small intestine).

[0269] Regardless of the type of dosage unit, it contains a therapeutically effective amount of one or more lipid compounds or the lipid composition described herein. One skilled in the art can determine a suitable amount of the compound to incorporate into the lipid composition, or pharmaceutical composition, in the dosage units.

[0270] In some embodiments, the lipid composition, pharmaceutical composition, or dosage unit contains about 0.01 to about 1000 mg of one or more lipid compounds described herein. In some embodiments, the lipid composition, pharmaceutical composition, or dosage unit contains about 0.01, about 0.1, about 0.5, about 1, about 5, about 10, about 25, about 50, about 75, about 100, about 125, about 150, about 175, about 200, about 225, 250, about 275, about 300, about 350, about 400, about 450, about 500, about 550, about 600, about 650, about 700, about 750, about 800, about 850, about 900, about 950, or about 1000 mg of one or more lipid compounds described herein. In some embodiments, the lipid composition, pharmaceutical composition, or dosage unit contains about 0.01 to about 750 mg, about 0.01 to about 500 mg, about 0.01 to about 250 mg, about 0.01 to about 100 mg, about 0.01 to about 50 mg, about 0.01 to about 25 mg, about 0.01 to about 10 mg, about 0.01 to about 5 mg, about 0.01 to about 0.1 mg, about 0.1 to about 1000 mg, about 0.1 to about 750 mg, about 0.1 to about 500 mg, about 0.1 to about 250 mg, about 0.1 to about 100 mg, about 0.1 to about 50 mg, about 0.1 to about 25, about 0.1 to about 10 mg, about 0.1 to about 5 mg, about 0.1 to about 1 mg, about 1 to about 1000 mg, about 1 to about 750 mg, about 1 to about 500 mg, about 1 to about 250 mg, about 1 to about 100 mg, about 1 to about 50 mg, about 1 to about 25 mg, about 1 to about 10 mg, about 1 to about 5 mg, about 5 to about 1000 mg, about 5 to about 750 mg, about 5 to about 500 mg, about 5 to about 250 mg, about 5 to about 100 mg, about 5 to about 50 mg, about 5 to about 25 mg, about 5 to about 10 mg, about 10 to about 1000 mg, about 10 to about 750 mg, about 10 to about 500, about 10 to about 250 mg, about 10 to about 100 mg, about 10 to about 50 mg, about 10 to about 25 mg, about 25 to about 1000 mg, about 25 to about 750 mg, about 25 to about 500 mg, about 25 to about 250 mg, about 25 to about 100 mg, about 25 to about 50 mg, about 50 to about 1000, mg about 50 to about 750 mg, about 50 to about 500 mg, about 50 to about 250 mg, about 50 to about 100 mg, about 100 to about 1000 mg, about 100 to about 750 mg, about 100 to about 500 mg, about 100 to about 250 mg, about 250 to about 1000 mg, about 250 to about 750 mg, about 250 to about 500 mg, about 500 to about 1000 mg, about 500 to about 750 mg, or about 750 to about 1000 mg of one or more lipid compounds described herein.- 84 - LEGAL\113833648\6Methods of Using the Lipid Composition

[0271] Also provided herein are methods for delivering a therapeutic agent to a cell or a subject (e.g., a patient) in need thereof, comprising contacting the cell with, or administering to the subject, the lipid composition according to any of the presently disclosed embodiments, or the pharmaceutical composition according to any of the presently disclosed embodiments, wherein the lipid composition comprises a therapeutic agent.

[0272] All above descriptions and all embodiments discussed in the above aspects relating to the novel lipid compounds, including the various genus and subgenus structural formulas, variable definitions and exemplary variables, and exemplary compounds, are all applicable to this aspect of the invention relating to the methods of delivering a therapeutic agent to a subject.

[0273] All above descriptions and all embodiments discussed in the above aspects relating to the lipid composition and pharmaceutical composition and formulation, including various other lipid components, the therapeutic agents, the pharmaceutically acceptable excipient, routes of administration and dosage forms and units, are applicable to these aspects of the invention relating to the methods of delivering a therapeutic agent to a subject.

[0274] In some embodiments, provided herein is a method of delivering the therapeutic agent to at least one cell, tissue, or organ, such as kidney, liver, splenic, lymphatic, or marrow of a subject in need thereof with a minimum amount delivered elsewhere in body of the subject. In some embodiments, the method delivers the therapeutic agent to the kidney or spleen of a subject in need thereof with a minimum amount delivered elsewhere in body of the subject.

[0275] It has surprisingly been discovered that the inventive lipid composition incorporating the lipid compound ((including one or more compounds of formula (GI), (GII), (GF), (GIF), (PE-I), or (PE -II); or any of the compounds belonging to any subgenus formulas or species under formula (GI), (GII), (GP), (GIF), (PE-I), or (PE-II) disclosed herein) preferentially targets kidney, liver, splenic, lymphatic, and / or marrow cells, and can thereby preferentially deliver the therapeutic agent (e.g., a protein or a nucleic acid) to such cells, tissues, or organs. In some embodiments, the therapeutic agent is delivered to kidney, liver, splenic, lymphatic, or marrow cells (including stem cells) of the subject. Such preferential delivery can occurs with or without the use of a specific targeting ligand.

[0276] In some embodiments, the lipid composition delivers therapeutic agent to kidney cells. In some embodiments, the delivery can preferentially target kidney cells. In some embodiments, the therapeutic agent is delivered to (e.g., by preferentially targeting) one or more of kidney endothelial cells, kidney epithelial cells, kidney dendritic cells, kidney B cells, kidney T cells, kidney natural killer (NK) cells, kidney macrophages, or other immune cells within the kidney,- 85 - LEGAL\113833648\6kidney collecting duct principal cells, kidney distal convoluted tubule cells, kidney glomerular endothelial cells, kidney intercalated cells, kidney interstitial fibroblasts, kidney mesangial cells, kidney parietal epithelial cells, kidney pericytes, kidney podocytes, kidney proximal tubule cells, kidney thick ascending limb cells, or kidney vascular endothelial cells.

[0277] In some embodiments, the lipid composition delivers therapeutic agent to liver cells. In some embodiments, the delivery can preferentially target liver cells. In some embodiments, the therapeutic agent is delivered to (e.g., by preferentially targeting) one or more of liver endothelial cells, hepatocytes, liver macrophages, liver dendritic cells, liver Kupffer cells, liver B cells, liver T cells, or other immune cells within the liver.

[0278] In some embodiments, the lipid composition delivers therapeutic agent to splenic cells. In some embodiments, the delivery can preferentially target splenic cells. In some embodiments, the therapeutic agent is delivered to (e.g., by preferentially targeting) one or more of spleen dendritic cells, spleen neutrophils, spleen macrophages, spleen B cells, spleen T cells, spleen natural killer (NK) cells, or other immune cells within the spleen.

[0279] In some embodiments, the lipid composition delivers therapeutic agent to lymphatic cells. In some embodiments, the delivery can preferentially target lymphatic cells. In some embodiments, the therapeutic agent is delivered to (e.g., by preferentially targeting) one or more of lymphatic dendritic cells, lymphatic neutrophils, lymphatic macrophages, lymphatic B cells, lymphatic T cells, or lymphatic natural killer (NK) cells of the subject.

[0280] In some embodiments, the lipid composition delivers therapeutic agent to marrow cells (including stem cells). In some embodiments, the delivery can preferentially target marrow cells (e.g., marrow stem cells). In some embodiments, the therapeutic agent is delivered to stem-like cells (e.g., bone marrow stem-like cells), and / or hematopoietic stem cells (HSCs) (e.g., bone marrow HSCs). In some embodiments, the therapeutic agent is delivered to (e.g., by preferentially targeting) one or more of marrow dendritic cells, marrow macrophages, marrow B cells, marrow T cells, other immune cells within marrow, or marrow stem-like cells, or marrow hematopoietic stem cells (HSCs).

[0281] In some embodiments, more than 99%, 95%, 90%, 85%, 80%, 75%, 70%, 65%, 60%, 55%, 50%, 45%, 40%, 35%, 30%, 25%, 20%, 15%, or 10% of the total therapeutic agent administered to the subject is delivered to the kidney, liver, splenic, lymphatic, or marrow cells of the subject. In some embodiments, more than 99%, 95%, 90%, 85%, 80%, 75%, 70%, 65%, 60%, 55%, 50%, 45%, 40%, 35%, 30%, 25%, 20%, 15%, or 10% of the total therapeutic agent administered to the subject is delivered to the kidney of the subject. In some embodiments, more than 99%, 95%, 90%, 85%, 80%, 75%, 70%, 65%, 60%, 55%, 50%, 45%, 40%, 35%, 30%, 25%, 20%, 15%, or 10% of the total therapeutic agent administered to the subject is delivered to the- 86 - LEGAL\113833648\6liver of the subject. In some embodiments, more than 99%, 95%, 90%, 85%, 80%, 75%, 70%, 65%, 60%, 55%, 50%, 45%, 40%, 35%, 30%, 25%, 20%, 15%, or 10% of the total therapeutic agent administered to the subject is delivered to the spleen of the subject. In some embodiments, more than 99%, 95%, 90%, 85%, 80%, 75%, 70%, 65%, 60%, 55%, 50%, 45%, 40%, 35%, 30%, 25%, 20%, 15%, or 10% of the total therapeutic agent administered to the subject is delivered to the marrow cells of the subject. In some embodiments, more than 99%, 95%, 90%, 85%, 80%, 75%, 70%, 65%, 60%, 55%, 50%, 45%, 40%, 35%, 30%, 25%, 20%, 15%, or 10% of the total therapeutic agent administered to the subject is delivered to the lymphatic cells of the subject.

[0282] As used herein, preferential delivery to a particular class of cells or cell type refers to delivery at a higher rate than to non-targeted cells. For example, the preferential delivery can mean delivery at or above a rate that is at least 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 20, 30, 40, 50, 60, 70, 80, 90, or 100 times greater than to non-targeted cells, i.e., cells not within the particular class of cells or of the particular cell type. In some embodiments, the delivery is to a particular targeted class of cells or cell type, and there is no delivery or only minimal delivery to non-targeted cells. As a result, the preferential delivery can be at a rate that is hundreds of times, thousands of times, or, theoretically infinitely greater than to the non-targeted cells.

[0283] Beneficially, the lipid composition can deliver the therapeutic agent to the subject at clinically relevant doses. In some embodiments, that dose at which the lipid composition delivers the therapeutic agent is about 0.005 to about 5.0 mg / kg. For example the dose at which the therapeutic agent is delivered may be about 0.005, 0.001, 0.002, 0.003, 0.004, 0.005, 0.006, 0.007, 0.008, 0.009, 0.01, 0.02, 0.03, 0.04, 0.05, 0.06, 0.07, 0.08, 0.09, 0.1, 0.11, 0.12, 0.13, 0.14, 0.15, 0.16, 0.17, 0.18, 0.19, 0.2, 0.21, 0.22, 0.23, 0.24, 0.25, 0.26, 0.27, 0.28, 0.29, 0.3, 0.35, 0.4, 0.45, 0.5, 0.55, 0.6, 0.65, 0.7, 0.75, 0.8, 0.85, 0.9, 0.95, 1.0, 1.1, 1.2, 1.3, 1.4, 1.5, 1.6, 1.7, 1.8, 1.9, 2.0, 2.1, 2.2, 2.3, 2.4, 2.5, 2.6, 2.7, 2.8, 2.9, 3.0, 3.1, 3.2, 3.3, 3.4, 3.5, 3.6, 3.7, 3.8, 3.9, 4.0, 4.1, 4.2., 4.3, 4.4, 4.5, 4.6, 4.7, 4.8, 4.9, or 5.0 mg / kg.

[0284] Accordingly, the disclosure also provides methods for delivering a therapeutic agent to a subject in need with specificity to kidney, liver, splenic, lymphatic, or marrow cells, tissues, or organs of a subject, the method comprising administering to the subject the lipid composition according to any of the presently disclosed embodiments, or the pharmaceutical composition according to any of the presently disclosed embodiments, wherein the lipid composition comprises a therapeutic agent.

[0285] Thus, in one aspect, provided is a method for delivering of a therapeutic agent to a subject with specificity to kidney, comprising: administering to the subject the lipid composition according to any of the presently disclosed embodiments, or the pharmaceutical composition according to any of the presently disclosed embodiments, wherein the lipid composition- 87 - LEGAL\113833648\6comprises a therapeutic agent. In some embodiments, the lipid compound in the lipid composition or the pharmaceutical composition is lipid No. 1, 8, 13, 17, 18, 26, 27, 32, 40, 45, 46, 50, 60, 88, 104, 106, 107, 123, 127, or 134. In some embodiments, the lipid compound in the lipid composition or the pharmaceutical composition is lipid No. 40, 26, 18, 45, 60, or 38.

[0286] In some embodiments, the therapeutic agent is preferentially delivered to kidney cells. In some embodiments, the therapeutic agent is delivered to (e.g., by preferentially targeting) one or more of kidney endothelial cells, kidney epithelial cells, kidney dendritic cells, kidney B cells, kidney T cells, kidney natural killer (NK) cells, kidney macrophages, or other immune cells within the kidney, kidney collecting duct principal cells, kidney distal convoluted tubule cells, kidney glomerular endothelial cells, kidney intercalated cells, kidney interstitial fibroblasts, kidney mesangial cells, kidney parietal epithelial cells, kidney pericytes, kidney podocytes, kidney proximal tubule cells, kidney thick ascending limb cells, or kidney vascular endothelial cells.

[0287] Also provided are methods of treating or preventing diseases or disorders in a subject in need thereof comprising administering to the subject the lipid composition according to any of the presently disclosed embodiments, or the pharmaceutical composition according to any of the presently disclosed embodiments. In some embodiments, the disease or disorder to be treated associated with expression or overexpression of a gene that can be reduced, decreased, downregulated, or silenced by the therapeutic agent delivered by the lipid composition or pharmaceutical composition.

[0288] Thus, in one aspect, provided is a method of treating a disease or disorder in a subject in need of such treatment, the method comprising: administering to the subject the lipid composition according to any of the presently disclosed embodiments, or the pharmaceutical composition according to any of the presently disclosed embodiments, wherein the lipid composition comprises a therapeutic agent.

[0289] In some embodiments, the disease or disorder is an infectious disease, cancer, a proliferative disease, a genetic disease, an autoimmune disease, diabetes, a neurodegenerative disease, a cardiovascular disease, a reno-vascular disease, or a metabolic disease.

[0290] In some embodiments, the disease or disorder may be cystic fibrosis, hemophilia A, hemophilia B, thalassemia, anemia and other blood disorders, AIDS, Alzheimer's disease, Parkinson's disease, Huntington's disease, amyotrophic lateral sclerosis, epilepsy, and other neurological disorders, cancer, diabetes mellitus, muscular dystrophies (e.g., Duchenne, Becker), Hurler's disease, adenosine deaminase deficiency, metabolic defects, retinal degenerative diseases (and other diseases of the eye), mitochondriopathies (e.g., Leber's hereditary optic neuropathy (LHON), Leigh syndrome, and subacute sclerosing encephalopathy), myopathies (e.g.,- 88 - LEGAL\113833648\6facioscapulohumeral myopathy (FSHD) and cardiomyopathies), diseases of solid organs (e.g., brain, liver, kidney, heart), and the like.

[0291] In some embodiments, the disease or disorder may be caused by mutation in a gene or gene product (i.e., a genetic disorder), such as metabolic diseases or disorders (e.g., Fabry disease, Gaucher disease, phenylketonuria (PKU), glycogen storage disease); urea cycle diseases or disorders (e.g., ornithine transcarbamoylase (OTC) deficiency); lysosomal storage diseases or disorders (e.g., metachromatic leukodystrophy (MLD), mucopolysaccharidosis Type II (MPSII; Hunter syndrome)); liver diseases or disorders (e.g., progressive familial intrahepatic cholestasis (PFIC); blood diseases or disorders (e.g., hemophilia (A and B), thalassemia, and anemia); cancers and tumors, and genetic diseases or disorders (e.g., cystic fibrosis).

[0292] In some embodiments, the genetic disorder is hemophilia A, hemophilia B, phenylketonuria (PKU, Gaucher disease Types I, II and III, Stargardt macular dystrophy, Leber congenital amaurosis (LCA), Usher syndrome, wet AMD.

[0293] Another aspect of the invention relates to a method for treating a kidney disease or kidney disorder, the method comprising: administering to the subject the lipid composition according to any of the presently disclosed embodiments, or the pharmaceutical composition according to any of the presently disclosed embodiments, wherein the lipid composition comprises a therapeutic agent. In some embodiments, the lipid compound in the lipid composition or the pharmaceutical composition is lipid No. 1, 8, 13, 17, 18, 26, 27, 32, 40, 45, 46, 50, 60, 88, 104, 106, 107, 123, 127, or 134. In some embodiments, the lipid compound in the lipid composition or the pharmaceutical composition is lipid No. 40, 26, 18, 45, 60, or 38.

[0294] In some embodiments, the kidney disease or kidney disorder is Alport syndrome, Bartter syndrome types 1-4, congenital nephrotic syndrome, cystinuria, cystinosis, Dent disease types 1 and 2, distal renal tubular acidosis, Fabry disease, familial amyloidosis, Gitelman syndrome, Liddle syndrome, Lowe syndrome, nephronophthisis (NPHP), autosomal dominant polycystic kidney disease, autosomal recessive polycystic kidney disease, tubulointerstitial kidney disease, proximal renal tubular acidosis, thin basement membrane disease, tubulointerstitial kidney disease, tuberous sclerosis complex (TSC), or APOL1 mediated kidney disease.EXAMPLES

[0295] The following examples are for illustrative purposes only and are not intended to limit, in any way, the scope of the present invention. To the extent that specific materials are mentioned, it is merely for illustrative purpose and is not intended to limit the invention. One- 89 - LEGAL\113833648\6skilled in the art may develop equivalent methods or reactants without the exercise of inventive capacity and without departing from the scope of the invention.Example 1 - General Reactions for the Synthesis of Ionizable Lipid CompoundsA. INDEX OF GENERAL PROCEDURES:

[0296] General procedure for ester synthesis - preparation of 3 -hexylnonyl 8-bromooctanoate (Intermediate I)EDC-HCIDMAP DIEA'OH DCMC23H45BrO23-hexylnonanol MW: 433.528-Br-octanoic acid

[0297] General procedure for secondary alcohol synthesis by flow - preparation of tridecan-5-ol (Intermediate II)nBuLi, hexanes ^..THF f-15 °C C13H28Ononanal MW: 200.37

[0298] General procedure for ester synthesis via acyl chlorides - preparation of 1 -butylnonyl 8-bromooctanoate (Intermediate III, alternate synthesis)C"’”'''-' o DIEA o HO '" „AC|..DCM C21H41BrO211 0 °C - RT MW: 405.46

[0299] General procedure for core amine monoalkylation - preparation of 1 -butylnonyl 8-[4-( / c / 7-butoxycarbonylamino) butylamino]octanoate (Intermediate IV)KI K2CO3MeCN A 1, HH3- RTC32H64N2O4 MW: 540.87IV

[0300] General procedure for second alkylation of core amine - preparation of 3 -hexylnonyl 8- [4-(tert-butoxycarbonylamino)butyl-[8-(l-butylnonoxy)-8-oxo-octyl]amino]octanoate (Intermediate V; Compound 3)- 90 - LEGAL\113833648\6KiKSGO3acetonitriteIll + iV80 °CCsaH-j 04N2O6MW: 865.42

[0301] General procedure for Boc cleavage followed by amide formation - preparation of 3-hexylnonyl 8-((8-oxo-8-(tridecan-5-yloxy)octyl)(4-(3- (phenylthio)propanamido)butyl)amino)octanoate (Compound 1)y1. HCI / dioxaneDCM2. EDC-HClDMAP DEA DCMC5ZHW4N2O5SMW: 929.53Compound 1

[0302] General procedure for benzyl ester hydrogenolysis by flow - preparation of 3-hexylnonyl 8-[[8-(l-butylnonoxy)-8-oxo-octyl]amino]octanoate (Intermediate VIII)Pd(OH)2 / AI2O3o o MeOH / THF / MeCN (flow reactor)C44H87NO4 VII MW: 694.18VHI

[0303] General procedure for Boc cleavage - preparation of 3-hexylnonyl 8-[4-aminobutyl-(8-benzyloxy-8-oxo-octyl)amino]octanoate, HC1 salt (Intermediate XXIII)0HC! 4M in EtOAc RTC^H?eN2O4. HCI MW: 673.08 (709 54)XXII XXiil

[0304] General procedure for benzyl ester hydrogenolysis over palladium on carbon -preparation of 8-[4-[3-(benzenesulfonyl)propanoylamino]butyl-[8-(l-butylnonoxy)-8-oxo-octyl]amino]octanoic acid, HCI salt (Intermediate XXIX)- 91 - LEGAL\113833648\6C44H7SN:, O?S. HO XXIV MW: 779.18 (815.64)XXIX

[0305] General procedure for headgroup amide coupling - preparation of 3-hexylnonyl 8-[4- [3-(benzenesulfonyl)propanoylamino]butyl-(8-benzyloxy-8-oxo-octyl)amino]octanoate (Intermediate XXIV; Compound 141)"bnxxut MW: 869.30XXIV

[0306] General procedure for sulfonyl chloride formation followed by sulfonamide formation - preparation of 3-hexylnonyl 8-((8-oxo-8-(tridecan-5-yloxy)octyl)(4-(N-(3- phenylpropyl)sulfamoyl)butyl)amino)octanoate (Compound 145)- HC57H1OSN2O0SMW; 947.54Compound 145- 92 - LEGAL\113833648\6

[0307] General procedure for Boc cleavage followed by sulfonamide formation - Preparation of 3 -hexylnonyl 8-((4-((2-(4-methoxyphenoxy)ethyl)sulfonamido)butyl)(8-oxo-8-(tridecan-5- yloxy)octyl)amino)octanoate (Compound 22)O 1. TFAA DC M0J02. NaHCOgX i0-Ji. N " -■ -N - - O - ' -0oc_RT**Et-NDCM0 °C - RTCompound 22

[0308] General procedure for sulfanyl ethanesulfonamide formation via Michael additionpreparation of 3-hexylnonyl 8-((4-((2-(benzylthio)ethyl)sulfonamido)butyl)(8-oxo-8-(tridecan-5- yloxy)octyl)amino)octanoate (Compound 649)O s'"'"', _ 6 „ i „. DSUS ' C>" - - _ ~OwO f o THF / DCM;o °c - RTHC57H106N2O6S2MW: 979.60Compound S9B. SYNTHESES OF INTERMEDIATES AND FINAL COMPOUNDS

[0309] General reaction schemes for synthesis of exemplary ionizable lipid compounds, derived from various intermediates, are shown in Schemes 1-5 below.- 93 - LEGAL\113833648\6B-l. Monoamine synthesis - Scheme 1: Synthesis of 3 -hexylnonyl 8-[4-( / c / 7-butoxycarbonyl amino)butyl-[8-(l -butylnonoxy)-8-oxo-octyl]amino]octanoate (Intermediate V)Scheme 1.EDC-HCIDMAP01 EA DCMnBuLi / hexanes THF-15 °C nonanal (flow reactor)EDC-HCI DMAP DIEA DCMKi KZCO3MeCN RTiy o Kl A K?CO3f '' acetonitrile i p p Uf + JV - *80 °CHC53HIO4N2O0 MW: 865.42Step 1: GENERAL PROCEDURE FORESTER SYNTHESIS - preparation of 3 -hexylnonyl 8-bromooctanoate (Intermediate I)EDC-HCI DMAP DIEA O DCMCjst'UgBrOj 3-hexylnonanol 8-Br-octanoic acid MW: 433.52

[0310] To a solution of 3-hexylnonan-l-ol (25.0 g, 109.4 mmol, 1.1 eq) and 8-bromooctanoic- 94 - LEGAL\113833648\6acid (22.2 g, 99.5 mmol, 1 eq) in DCM (220 mL) was added / V. / V-di isopropyl ethylamine (DIEA; 38.6 g, 298.5 mmol, 52.0 mL, 3 eq), 4-(dimethylamino)pyridine (DMAP; 1.22 g, 9.95 mmol, 0.1 eq) and 1 -ethyl-3 -(3 -dimethylaminopropyl)carbodiimi de hydrochloride (EDC-HC1; 28.6 g, 149.3 mmol, 1.5 eq) with stirring under a dry N2 atmosphere. The reaction was stirred at 20 °C for 12 hr, the reaction mixture poured into water (500 mL) and extracted with ethyl acetate (EtOAc; 3x 500 mL). The combined organic phase was washed with brine (2x 200 mL), dried over anhydrous Na2SC>4, fdtered and the filtrate concentrated in vacuo. The residue was purified by silica gel chromatography, eluting with 100% hexanes going to 90% hexanes / 10% EtOAc to give 30 g (69.20 mmol, 69.6% yield) of the title compound as a colorless oil. 'H-NMR (400MHZ, MeOD, d): 4.11 (t, J = 6.6 Hz, 2H), 3.44 (t, J= 6.6 Hz, 2H), 2.31 (t, J= 7.4 Hz, 2H), 1.85 (m, 2H), 1.65-1.55 (m, 4H), 1.52-1.40 (m, 3H), 1.39-1.25 (m, 24H), 0.91 (m, 6H).Step 2: GENERAL PROCEDURE FOR SECONDARY ALCOHOL SYNTHESIS BY FLOW -preparation of tridecan-5-ol (Intermediate II)nBuli / hexanesTHF (-15 X C13H28O. (Sow reactor) MW: 200.37nonanalII

[0311] Flow chemistry: Solution 1: nonanal (120 g, 843.7 mmol, 1 eq) in 1200 mL THF. Solution 2: n-BuLi (1.75 eq, 1.6 M in hexanes, 925 mL). Pump 1 (SI, Pl, 70.5 mL / min) introduced solution 1 to flow reactor 1 (FLR1, PFA, Coils reactor, 3.175(1 / 8") mm, 29.957 mL, -15 °C). Pump 2 (S2, P2, 49.3 mL / min) introduced solution 2 to flow reactor 1 (FLR1). Pumps 1 and 2 were started at the same time, the residence time of flow reactor 1 was 0.25 min. and the product mixture was collected in a flask as it exited the reactor. After 90 minutes the obtained product mixture was poured into water (500 mL) and extracted with ethyl acetate (3x 500 mL). The combined organic phase was washed with brine (2x 500 mL), dried with anhydrous Na2SC>4, filtered and the filtrate was concentrated in vacuo to give a residue. The residue was purified by silica gel chromatography, eluting with petroleum ether 100% going to 99% pet. ether / 1% EtOAc to give 80 g (399.3 mmol, 50% yield) of the title compound as a colorless oil. 'H-NMR (400MHz, MeOD, d): 3.50 (br. t, J= 4.0 Hz, 1H), 1.45-1.38 (m, 6H), 1.37-1.25 (m, 15H), 0.95 0.85 (m, 6H).- 95 - LEGAL\113833648\6Step 3: Preparation of 1 -butylnonyl 8-bromooctanoate (Intermediate III)EDC-HCI X-.. „ DMAP- MW: 405.46 IB

[0312] Following the general procedure for ester synthesis using tridecan-5-ol (Intermediate II; 25 g, 124.8 mmol, 1.5 eq) in place of 3-hexylnonan-l-ol and 8-bromooctanoic acid (18.56 g, 83.2 mmol, 1 eq) in DCM (250 mL) were added EDC-HCI (17.54 g, 91.5 mmol, 1.1 eq), DIEA (32.25 g, 249.5 mmol, 43.5 mL, 3 eq) and DMAP (1.02 g, 8.32 mmol, 0.1 eq) with stirring under a dry N2 atmosphere. The reaction was stirred at 20 °C for 10 hr, the reaction mixture reduced in vacuo, poured into water (200 mL) and extracted with ethyl acetate (EtOAc; 3x 500 mL). The combined organic phase was washed with brine (2x 200 mL), dried over anhydrous Na2SC>4, filtered and the filtrate concentrated in vacuo. The residue was purified by silica gel chromatography, eluting with 100% petroleum ether going to 99% pet. ether / 1% EtOAc to give 11 g (27.1 mmol, 32.6% yield) of the title compound as a colorless oil.1H-NMR (400MHz, CDCI3, d): 4.88 (m, 1H), 3.41 (t, J= 7.0 Hz, 2H), 2.31 (t, J= 7.2 Hz, 2H), 1.92-1.86 (m, 2H), 1.52 (m, 2H), 1.50-1.40 (m, 6H), 1.39-1.22 (m, 20H), 0.89 (m, 6H).Step 4: GENERAL PROCEDURE FOR CORE AMINE MONO ALKYLATION - preparation of 1 -butylnonyl 8-[4-( / c / 7-butoxy carbonylamino) butylamino]octanoate (Intermediate IV)KICMH64N2O4i MW: 540.87iy

[0313] To a mixture of 1 -butylnonyl 8-bromooctanoate (Intermediate III; 10 g, 24.7 mmol, 1 eq) and tert-butyl A-(4-aminobutyl (carbamate (9.29 g, 49.3 mmol, 2 eq) in acetonitrile (MeCN; 100 mL) was added K2CO3 (10.23 g, 74.0 mmol, 3 eq) and KI (409.4 mg, 2.47 mmol, 0.1 eq) with stirring under a dry N2 atmosphere. The reaction was stirred at 20 °C for 12 hr, the reaction mixture reduced in vacuo, poured into water (500 mL) and extracted with EtOAc (3x 500 mL). The combined organic phase was washed with brine (2x 200 mL), dried over anhydrous Na2SO4, filtered and the filtrate concentrated in vacuo. The residue was purified by silica gel chromatography, eluting with 100% DCM going to 90% DCM / 10% MeOH to give 30 g (58.50 mmol, 47.44% yield) of the title compound as a yellow oil. 'H-NMR (400MHz, CDCI3, d): 4.92- 4.84 (m, 2H), 3.12 (d, J= 4.8 Hz, 2H), 2.64-2.55 (m, 3H), 2.29 (t, J= 7.2 Hz, 2H), 1.69-1.56 (m,- 96 - LEGAL\113833648\62H), 1.53-1.40 (m, 21H), 1.39-1.20 (m, 22H), 0.89 (m, 6H).Step 5: GENERAL PROCEDURE FOR SECOND ALKYLATION OF CORE AMINE -preparation of 3-hexylnonyl 8-[4-( / c / 7-butoxycarbonylamino)butyl-[8-( I -butylnonoxy)-8-oxo-octyl]amino]octanoate (Intermediate V; Compound 3)KI A AK2CO3I" "0' - ''acetonitrile < o otil + iv - *■ 4 A N. A A „-x.- 80l!CH^53Ff04N2O6MW: 865.42V

[0314] To a solution of 3-hexylnonyl 8-bromooctanoate (Intermediate I, 10.99 g, 25.35 mmol, 1.3 eq) and 1-butylnonyl 8-[4-( / c / 7-butoxycarbonylamino)butylamino]octanoate (Intermediate IV; 10 g, 19.50 mmol, 1 eq) in MeCN (100 mL) was added K2CO3 (8.09 g, 58.50 mmol, 3 eq) and KI (323.71 mg, 1.95 mmol, 0.1 eq) with stirring under a dry N2 atmosphere. The reaction was stirred at 80 °C for 12 hr, the reaction mixture reduced in vacuo, poured into water (500 mL) and extracted with EtOAc (3x 500 mL). The combined organic phase was washed with brine (2x 200 mL), dried over anhydrous Na2SC>4, fdtered and the fdtrate concentrated in vacuo. The residue was purified by silica gel chromatography, eluting with 100% petroleum ether going to 75% pet. ether / 25% EtOAc to give 26 g (30.0 mmol, 51.4% yield) of the title compound as a pale yellow oil. UPLC-MS: m / z (ESI) for C53H104N2O6 [M+H]+expected:865.80; observed: 865.9; 'H-NMR (400MHz, CDCh, d): 4.88 (s, 1H), 4.11 (t, J= 6.8 Hz, 2H), 3.05 (t, J= 6.4 Hz, 2H), 2.45 (m, 6H), 2.31 (t, J= 7.2 Hz, 4H), 1.68-1.56 (m, 3H), 1.55-1.50 (m, 6H), 1.49-1.40 (m, 18H), 1.39-1.23 (m, 50H), 0.91 (m, 12H).- 97 - LEGAL\113833648\6B-2. Monoamine synthesis - Scheme 2: Synthesis of 3 -hexylnonyl 8-[[8-(l-butylnonoxy)-8-oxo-octyl]amino]octanoate (Intermediate VIII)Scheme 2.8nNH2KIK2CO3MeCNHRTli:Intermediate IKIK?CO:iacetonitrile80 °CH, Pd(OH)j / AI^O3MeOH / THF / MeCN(flow reactor)VIIIStep 1: Preparation of 1 -butylnonyl 8-(benzylamino)octanoate (Intermediate VI)SnNH>:KI-x K; CO,RTC28H49NO2W MW: 431.71

[0315] Following the general procedure for core amine monoalkylation, benzylamine (43.61 g, 406.9 mmol, 44.4 mL, 5 eq) was reacted with 1-butylnonyl 8-bromooctanoate (Intermediate III; 33 g, 81.4 mmol, 1 eq) followed by an aqueous workup. The residue was purified by silica gel chromatography, eluting with 100% hexanes going to 75% hexanes / 25% EtOAc to give 20 g (46.3 mmol, 56.9% yield) of the title compound as a colorless oil.1H-NMR (400MHz, MeOD, d): 7.34-7.22 (m, 5H), 4.88 (m, 1H; partially obscured by HOD peak), 3.73 (s, 2H), 3.56 (t, J= 7.4 Hz, 2H), 2.29 (t, J= 7.2 Hz, 2H), 1.67- 1.49 (m, 8H), 1.41-1.25 (m, 22H), 0.90 (m, 6H); amine proton not observed.- 98 - LEGAL\113833648\6Step 2: Preparation of 3 -hexylnonyl 8-[benzyl-[8-(l-butylnonoxy)-8-oxo-octyl]amino]octanoate (Intermediate VII)Intermediate IKIK?:CO3acetonitrile 80 °CMW: 784.31

[0316] Following the general procedure for second alkylation of core amine, 1 -butylnonyl 8-(benzylamino)octanoate (Intermediate VI; 15 g, 34.75 mmol, 1 eq) was reacted with 3-hexylnonyl 8-bromooctanoate (Intermediate I; 18.1 g, 41.7 mmol, 1.2 eq) followed by an aqueous workup. The residue was purified by silica gel chromatography, eluting with 100% hexanes going to 90% hexanes / 10% EtOAc to give 16 g, (20.4 mmol, 58.7% yield) of the title compound as a colorless oil. 'H-NMR (400MHz, CDCL, d): 7.35-7.20 (m, 5H; partially obscured by solvent peak), 4.87 (m, 1H), 4.08 (t, J= 7.2 Hz, 2H), 3.53 (s, 2H), 2.38 (t, J= 7.4 Hz, 2H), 2.27 (t, J= 7.6 Hz, 2H), 1.65- 1.36 (br. m, 18H), 1.34-1.20 (m, 47H), 0.89 (m, 12H)Step 3: GENERAL PROCEDURE FOR BENZYLAMINE HYDROGENOLYSIS BY FLOW -Preparation of 3 -hexylnonyl 8-[[8-(l-butylnonoxy)-8-oxo-octyl]amino]octanoate (Intermediate VIII) H2Pd{OH)2 / AI203MeOH / THFTMeCN (flow reactor) Vil

[0317] Flow chemistry: Solution 1: 3-hexylnonyl 8-[benzyl-[8-(l-butylnonoxy)-8-oxo-octyl]amino]octanoate (Intermediate VII; 15 g, 19.13 mmol, 1 eq) in 330 mL of a THF / MeOH / MeCN 5:5:1 mixture. The fixed bed (named FLR1, volume 5 mL) was completely packed with granular catalyst 5% Pd(OH)2 / AI2O3 (WXC1055, 4 g). The H2 back pressure regulator was adjusted to 3 MPa, and the flow rate of H2 was 60 mL / min. Then the solution SI was pumped by Pump 1 (SI, Pl, 0.3 mL / min) to fixed bed FLR1 (SS, fixed bed, 6.350 (1 / 4”) mm, 1 mL, 100 °C) with a 3.3 min residence time. The product mixture was collected in a flask as it exited the reactor and concentrated in vacuo to give a colorless oil. This was partitioned between EtOAc (200 mL) and sat. Na2COs (200 mL). The organic phase was separated and the aqueous phase extracted with ethyl acetate (2x 100 mL). The combined organic phase was washed with brine (2x 100 mL), dried over anhydrous Na2SO4, filtered and the filtrate- 99 - LEGAL\113833648\6concentrated. The residue was purified by silica gel chromatography, eluting with 100% hexanes going to 80% hexanes / 20% EtOAc to give 10 g (14.4 mmol, 75.3% yield) of the title compound as yellow oil. UPLC-MS: m / z (ESI) for C44H87NO4 [M+H]+expected: 694.67; observed: 694.8; 'H-NMR (400MHz, CDCh, d): 4.87 (m, 1H), 4.08 (t, J= 7.2 Hz, 2H), 2.74 (m, 2H), 2.28 (t, J = 7.4 Hz, 2H), 1.70- 1.51 (br. m, 18H), 1.36-1.20 (m, 49H), 0.89 (m, 12H)B-3. Monoamine synthesis - Scheme 3: Synthesis of 4-[[8-(l-butylnonoxy)-8-oxo-octyl]-[8-(3-hexylnonoxy)-8-oxo- octyl]amino]butane-l-sulfonic acid (Intermediate IX)Scheme 3.MW: 830.35 IX

[0318] A mixture of 3 -hexylnonyl 8-[[8-(l-butylnonoxy)-8-oxo-octyl]amino]octanoate (Intermediate VIII; 9 g, 12.97 mmol, 1 eq) and 1,4-butane sultone (8.83 g, 64.8 mmol, 6.63 mL, 5 eq) in acetone (90 mL) was stirred at 80 °C for 48 h under dry N2. The reaction mixture was concentrated and the residue purified by silica gel chromatography, eluting with 100% dichloromethane going to 90% DCM / 10% MeOH to give 5.1 g (6.14 mmol, 47.4% yield) of the title compound as a colorless oil. UPLC-MS: m / z (ESI) for C48H95NO7S [M+H]+expected:830.69; observed: 830.8. 'H-NMR (400MHz, CDCh, d): 10.55 (s, 1H), 4.87 (m, 1H), 4.08 (t, J = 7.2 Hz, 2H), 3.14-2.94 (m, 8H), 2.29 (m, 4H), 2.08-1.89 (m, 4H), 1.84- 1.65 (br. m, 4H), 1.63-1.45 (m, 11H), 1.15 (m, 48), 0.89 (m, 12H)B-4. Monoamine synthesis - Scheme 4R: Synthesis of (5 / ?)-tridecan-5-ol (Intermediate XII) Scheme 4R.cuiTHFCuiTHF- 100 - LEGAL\113833648\6Step 1: Preparation of (2 )-l-chlorodecan-2-ol (Intermediate X)ocm,.. MgBi _ _, HTHF L:(PORT CWH21CIOMW: 192.73X

[0319] To a solution of (6)-epichlorohydrin (46.3 g, 500.0 mmol, 39.2 mL, 1 eq) in THF (380 mL) was added Cui (9.52 g, 50.0 mmol, 0.1 eq) with stirring to give a suspension. The mixture was cooled to 0 °C under a dry N2 atmosphere, then w-heptylmagnesium bromide (1 M, 600 mL, 1.2 eq) was added slowly with stirring, maintaining a temperature between 0-15°C. After addition, the mixture was allowed to warm to 25 °C and stirred for 12 h. The reaction was quenched with sat. aqueous NH4CI (500 mL) and the mixture stirred for 30 min, then diluted with EtOAc (500 mL). The organic phase was separated and the aqueous phase extracted with EtOAc (3x 200 mL). The organic layers were combined, washed with brine (2x 100 mL), dried over anhydrous NazSCL, fdtered and the filtrate concentrated in vacuo. The residue was purified by silica gel chromatography, eluting with 100% hexanes going to 90% hexanes / 10% EtOAc to give 40 g (207.5 mmol, 49.8% yield) of the title compound as a white solid. 'H-NMR (400MHZ, CDCh, d): 3.81 (br s, 1H), 3.68-3.62 (m, 1H), 3.51-3.46 (m, 1H), 2.12 (br. s, 1H), 1.59-1.44 (m, 4H), 1.43-1.21 (m, 10H), 0.89 (t, J = 7.2 Hz, 3H).Step 2: Preparation of (2X)-2-octy I oxirane (Intermediate XI)KI KICO3I 'PrOH80 °C C10H20OxMW: 156.27Xi

[0320] To a solution of (26)-l-chlorodecan-2-ol (Intermediate X, 40 g, 207.5 mmol, 1 eq) in 2-propanol (400 mL) was added K2CO3 (57.4 g, 415.1 mmol, 2 eq) and KI (3.45 g, 20.75 mmol, 0.1 eq) with stirring under dry N2. The mixture was heated to 80 °C and stirred for 12 h, then allowed to cool to room temp. The reaction was quenched with sat. aqueous NH4CI (1 L) and stirred for 30min, then diluted with EtOAc (1 L). The organic phase was separated and the aqueous phase extracted with EtOAc (3x 500 mL). The organic layers were combined, washed with brine (2x 200 mL), dried over anhydrous Na2SO4, filtered, and the filtrate concentrated. The residue was purified by silica gel chromatography, eluting with 100% hexanes going to 98%- 101 - LEGAL\113833648\6hexanes / 2% EtOAc to give 18 g (115.2 mmol, 55.5% yield) of the title compound as a pale yellow oil. 'H-NMR (400MHz, CDCh, d): 2.91 (m, 1H), 2.75 (t, J= 4.4 Hz, 1H), 2.47 (m, 1H), 1.52 (m, 2H; partially obscured by water peak), 1.49-1.39 (m, 2H), 1.38-1.20 (m, 10H), 0.90 (m, 3H).Step 3: Preparation of / 5 / / >-tridecan-5-ol (Intermediate XII)vTHF0 °C - RT „uY< CI3H28O™ MW: 200.37XII

[0321] To a solution of (2 )-2-octyloxirane (Intermediate XI; 18 g, 115.2 mmol, 1 eq) in THF (180 mL) was added Cui (2.19 g, 11.5 mmol, 0.1 eq) with stirring to give a suspension. The mixture was cooled to 0 °C under dry N2 and w-propylmagnesium bromide (2 M, 69.1 mL, 1.2 eq) was added slowly with stirring, maintaining a temperature between 0-15°C. After addition, the mixture was allowed to warm to 25 °C and stirred for 12 h. The reaction was quenched with sat. aqueous NH4CI (500 mL), the mixture stirred for 30 min, then diluted with EtOAc (500 mL). The organic phase was separated and the aqueous phase extracted with EtOAc (3x 300 mL). The organic layers were combined, washed with brine (2x 100 mL), dried over anhydrous Na2SO4, fdtered, and the filtrate concentrated in vacuo. The residue was purified by silica gel chromatography, eluting with 100% hexanes going to 97% hexanes / 3% EtOAc to give 10 g (49.9 mmol, 43.3% yield) of the title compound as a white solid. 'H-NMR (400MHZ, CDCL, d): 3.59 (m, 1H), 1.59-1.38 (m, 8H), 1.37-1.20 (m, 12H), 0.90 (m, 6H). Enantiomeric excess (%ee) by chiral GC = 93.7% [Column: Agilent J& W CP-Chirasil Dex CB 25 m*0.25 mm ID*0.25 pm film thickness (P / N: CP7502); Injector Temperature: 240 °C; Split Ratio: 30:1; Control Mode: pressure; Flow: 1.0 mL / min.; carrier gas: N2; Detector: Flame Ionization Detector (FID); Detector Temperature: 240 °C; Makeup (He) Flow: 15 mL / min.; H2 Flow: 40 mL / min.; Air Flow: 400 mL / min.; Program: Rate: 30 °C / min; 110 °C, hold time 90 min., 200 °C, hold time 2 min.,; GC Run Time: 95 min.]- 102 - LEGAL\113833648\6B-5. Monoamine synthesis - Scheme 4S: Synthesis of (5 )-tridecan-5-ol (Intermediate XV) Scheme 4S.,, MgBrMgBrK^CO3, KIi-PrOHStep 1: Preparation of (2 / ?)-l-chlorodecan-2-ol (Intermediate XIII)MgBrTHF0 °C - RT C10Hz1CiOMW: 192.73

[0322] Reaction was performed as in Step 1 in Scheme 4Rbut using (R)-epichlorohydrin (23.1 g, 250 mmol, 19.6 mL, 1 eq), Cui (4.76 g, 25.0 mmol, 0.1 eq), and w-heptylmagnesium bromide (1 M, 300 mL, 1.2 eq). Using the same aqueous workup and silica gel chromatography conditions gave 35 g (145.3 mmol, 58.1% yield) of the title compound as a yellow oil. 'H-NMR (400MHZ, CDCL, d): 3.81 (br s, 1H), 3.68-3.61 (m, 1H), 3.52-3.46 (m, 1H), 2.14 (br. s, 1H), 1.61-1.41 (m, 4H), 1.39-1.21 (m, 10H), 0.89 (t, J= 6.8 Hz, 3H).Step 2: Preparation of (2 / ?)-2-octyloxirane (Intermediate XIV)'PrOH80cCMW: 156.27XiV

[0323] Reaction was performed as in Step 2 in Scheme 4Rbut using U7J-l-chlorodecan-2-ol (Intermediate XIII; 35 g, 145.3 mmol, 1 eq), K2CO3 (40.2 g, 290.6 mmol, 2 eq) and KI (2.41 g, 14.5 mmol, 0.1 eq) in 350 mL 2-propanol. Following an aqueous workup the residue was purified with silica gel chromatography, eluting with 100% DCM going to 99% DCM / 1% MeOH, to- 103 - LEGAL\113833648\6give 14 g (89.6 mmol, 61.7% yield) of the title compound as a yellow oil. 'H-NMR (400MHZ, CDCh, d): 2.91 (m, 1H), 2.75 (t, J= 4.6 Hz, 1H), 2.47 (m, 1H), 1.57-1.42 (m, 4H), 1.37-1.20 (m, 10H), 0.89 (m, 3H).Step 3: Preparation of 5 )-tridecan-5-ol (Intermediate XV)CuiV-o. OHTHF0UC - RTC13H28OXIV MW: 200.37XV

[0324] Reaction was performed as in Step 3 in Scheme 4R but using (27^-2-octy 1 oxirane (Intermediate XIV; 14 g, 89.6 mmol, 1 eq), Cui (1.71 g, 8.96 mmol, 0.1 eq), and n- propylmagnesium bromide (2 M, 53.8 mL, 1.2 eq). Following an aqueous workup the residue was purified by silica gel chromatography, eluting with 100% hexanes going to 98% hexanes / 2% EtOAc to give 7 g (34.9 mmol, 39.0% yield) of the title compound as a white solid. 'H-NMR (400MHz, CDCh, d): 3.59 (m, 1H), 1.59-1.38 (m, 8H), 1.37-1.20 (m, 12H), 0.90 (m, 6H).Enantiomeric excess (%ee) by chiral GC = 93.4% [conditions - see Intermediate XII],B-6. Monoamine synthesis - Scheme 5R: Synthesis of 3 -hexylnonyl (K -8-((4-((tert- butoxycarbonyl)amino)butyl)(8-oxo-8-(tridecan-5-yloxy)octyl)amino)octanoate (Intermediate XVII)Scheme 5R.EDC-HCIo DMAP6r,.. 11 01 EA' 'OHDCM XII KIK3CO3acetonitrileBr- ‘O’ IV >80nCXVI- 104 - LEGAL\113833648\6XVHStep 1: Preparation of (lR)-\ -butylnonyl 8-bromooctanoate (Intermediate XVI)EDC-HCInDMAP y niFABr0 '.A'_xDCM °,k:C2-|H4iBrO2— MW: 405.46XVI

[0325] Reaction was performed as for Intermediate III but using (5 / ?)-tridecan-5-ol (Intermediate XII; 6 g, 29.9 mmol) in place of racemic tri decan- 5 -ol. Following aqueous workup, the material was purified by silica gel chromatography, eluting with 100% hexanes going to 98% hexanes / 2% EtOAc to give 8.5 g (21.0 mmol, 70% yield) of the title compound as a pale yellow oil. 'H-NMR (400MHz, CDC13, d): 4.87 (m, 1H), 3.41 (t, J= 7.0 Hz, 2H), 2.30 (t, J= 7.2 Hz, 2H), 1.92-1.86 (m, 2H), 1.52 (m, 2H), 1.50-1.40 (m, 6H), 1.39-1.22 (m, 20H), 0.89 (m, 6H).Step 2: Preparation of 3 -hexylnonyl <7 -8-((4-(( / c / 7-butoxycarbonyl)amino)butyl)(8-oxo-8-(tridecan-5-yloxy)octyl)amino)octanoate (Intermediate XVII)Ki,x - A. 1K. CG; [ " ""0" "acetonitrile: O R o V''--''XVI + IV - - A80 °C 'MC53H104N2O6MW: 865.42XVH

[0326] Reaction was performed as for Intermediate V using (lR)-\ -butylnonyl 8-bromooctanoate (Intermediate XVI; 8.28 g, 20.4 mmol) in place of racemic 1 -butylnonyl 8-bromooctanoate. Following aqueous workup, the material was purified by silica gel chromatography, eluting with 100% hexanes going to 80% hexanes / 20% EtOAc to give 5.1 g (5.89 mmol, 37.6% yield) of the title compound as a pale yellow oil. UPLC-MS: m / z (ESI) for C53H104N2O6 [M+H]+expected:865.80; observed: 866.0. 'H-NMR (400MHz, CDCh, d): 4.88 (s, 1H), 4.11 (t, J= 6.8 Hz, 2H), 3.05 (t, J= 6.4 Hz, 2H), 2.45 (m, 6H), 2.31 (t, J= 7.2 Hz, 4H), 1.68-1.56 (m, 3H), 1.55-1.50 (m, 6H), 1.49-1.40 (m, 18H), 1.39-1.23 (50H), 0.91 (m, 12H).- 105 - LEGAL\113833648\6B-7. Monoamine synthesis - Scheme 5S: Synthesis of 3 -hexylnonyl (S)-8- (4- (tert-butoxycarbonyl)amino)butyl)(8-oxo-8-(tridecan-5-yloxy)octyl)amino)octanoate (Intermediate XIX)Scheme 5S.EDC-HCIDMAP DIEA DCMKIK2CO:i9 acetonitrile+fv - »80l;CXVHIXIXStep 1: Preparation of (1S)-1 -butylnonyl 8-bromooctanoate (Intermediate XVIII)EDC-HCI DMAP DIEA. A X DCM O-;sA'C2iH41BrO XV2MW: 405.46XVIII

[0327] Reaction was performed as for Intermediate III using (5X)-tridecan-5-ol (Intermediate XV; 7 g, 34.9 mmol) in place of racemic tridecan-5-ol. Following aqueous workup, the material was purified by silica gel chromatography, eluting with 100% hexanes going to 98% hexanes / 2% EtOAc to give 8.5 g (21.0 mmol, 70% yield) of the title compound as a pale yellow oil.1H-NMR (400MHz, CDCh, d): 4.87 (m, 1H), 3.41 (t, J= 7.0 Hz, 2H), 2.30 (t, J= 7.2 Hz, 2H), 1.92-1.86 (m, 2H), 1.52 (m, 2H), 1.50-1.40 (m, 6H), 1.39-1.22 (m, 20H), 0.89 (m, 6H).- 106 - LEGAL\113833648\6Step 2: Preparation of 3 -hexylnonyl 4S)-8-((4-(( / c / 7-butoxycarbonyl)amino)butyl)(8-oxo-8-(tridecan-5-yloxy)octyl)amino)octanoate (Intermediate XIX)KI,.. E,,, KJCO3[0acetonitrile: o r' o »''XVIH + IV - ►80 °C 'HC53H1£MN2O6MW: 865.42X! X

[0328] Reaction was performed as for Intermediate V using (lS)- -butylnonyl 8-bromooctanoate (Intermediate XVIII; 9 g, 22.2 mmol) in place of racemic 1 -butylnonyl 8-bromooctanoate. Following aqueous workup, the material was purified by silica gel chromatography, eluting with 100% hexanes going to 80% hexanes / 20% EtOAc to give 5.3 g (6.12 mmol, 27.6% yield) of the title compound as a yellow oil. UPLC-MS: m / z (ESI) for C53H104N2O6 [M+H]+expected:865.80; observed: 866.0.XH-NMR (400MHz, CDCh, d): 4.88 (s, 1H), 4.11 (t, J= 6.8 Hz, 2H), 3.05 (t, J= 6.4 Hz, 2H), 2.45 (m, 6H), 2.31 (t, J= 7.2 Hz, 4H), 1.68-1.56 (m, 3H), 1.55-1.50 (m, 6H), 1.49-1.40 (m, 18H), 1.39-1.23 (50H), 0.91 (m, 12H).B-8. Monoamine synthesis - Scheme 6: Synthesis of benzyl 8-((8-((3-hexylnonyl)oxy)-8-oxooctyl)(4-(3 -(phenylsulfonyl)propanamido)butyl)amino)octanoate (Intermediate XXIV) Scheme 6.BnOHDCC OMAPDfEA o > oJI,+1.11,DCM OBn * q '0' N!HXXIntermediate IKt KIK2CO3K2CO3MeCN oJI acetonitrile OBr> >RT 80 °CHC!4M in EtOAcRTXXii- 107 - LEGAL\113833648\6or 9 f ' + fl _ tI.N- ^•••' X " -OH0 OXXII iEDC-HCI ODMAPEbN J......-: _ - 0 ( O A '■'•DCMO' '0HXX iVStep 1: Preparation of benzyl 8-bromooctanoate (Intermediate XX)BnOHEDC-HCIDMAP DIEA DCMMW: 313.23XX

[0329] To a mixture of 8-bromooctanoic acid (100 g, 448.2 mmol, 1 eq) and benzyl alcohol (48.5 g, 448.2 mmol, 46.4 mL, 1 eq) in DCM (1000 mL) was added DMAP (5.48 g, 44.82 mmol, 0.1 eq) in one portion with stirring at 20 °C under dry N2. A solution of dicyclohexylcarbodiimide (DCC; 101.7 g, 493.0 mmol, 99.7 mL, 1.1 eq) in DCM (500 mL) was added to the solution slowly. The mixture was stirred at 20 °C for 10 hours, poured into water (1000 mL) and extracted with DCM (3x 500 mL). The combined organic phase was washed with brine (2x 500 mL), dried over anhydrous Na2SO4, filtered and the filtrate concentrated in vacuo. The residue was purified by silica gel chromatography, eluting with 100% petroleum ether going to 98% pet. ether / 2% EtOAc to give 95 g, (303.3 mmol, 67.7% yield) of the title compound as a colorless oil. 'H-NMR (400MHz, MeOD, d): 7.36-7.30 (m, 5H), 5.12 (s, 2H), 3.41 (t, J= 6.8 Hz, 2H), 2.36 (t, J= 7.4 Hz, 2H), 1.81 (m, 2H), 1.63 (m, 4H), 1.42 (m, 2H), 1.32 (m, 4H).Step 2: Preparation of benzyl 8-[4-( / e / 7-butoxycarbonylamino)butylamino]octanoate (Intermediate XXI)KI K?CO.j MeCN >RTC24H40N2O4MW: 420.59XXI

[0330] To a mixture of benzyl 8-bromooctanoate (Intermediate XX; 45 g, 143.7 mmol, 1 eq) and tert-butyl A-(4-aminobutyl)carbamate (54.1 g, 287.3 mmol, 2.0 eq) in acetonitrile (500 mL)- 108 - LEGAL\113833648\6was added K2CO3 (59.6 g, 431.0 mmol, 3 eq) and KI (2.38 g, 14.4 mmol, 0.1 eq) in one portion and the reaction stirred at 20 °C under dry N2 for 10 hours. The mixture was fdtered and the filtrate concentrated in vacuo. The residue was purified by silica gel chromatography, eluting with 100% hexanes going to 100% EtOAc to give 42 g, (99.9 mmol, 34.7% yield) of the title compound as a yellow oil.XH-NMR (400MHz, CDCh, d): 7.37-7.32 (m, 5H), 5.11 (s, 2H), 4.93 (br s, 1H), 3.12 (br d, J = 4.8 Hz, 2H), 2.61-2.54 (m, 4H), 2.35 (t, J= 7.6 Hz, 2H), 1.51 (m, 2H), 1.47 (m, 4H), 1.44 (m, 12H), 1.31 (m, 6H).Step 3: Preparation of 1-3 -hexylnonyl 8-[(8-benzyloxy-8-oxo-octyl)-[4-(tert-butoxycarbonylamino)butyl]amino]octanoate (Intermediate XXII)Intermediate I KI K2CO3acetonitrile 80 °C C^7HB4N2OHMW: 773.20

[0331] Following the general procedure for second alkylation of core amine benzyl 8-[4-(tert-butoxycarbonylamino)butylamino]octanoate (Intermediate XXI; 20 g, 47.6 mmol, 1 eq) was reacted with 3-hexylnonyl 8-bromooctanoate (Intermediate I; 24.7 g, 57.1 mmol, 1.2 eq) followed by an aqueous workup. The residue was purified by silica gel chromatography, eluting with 100% hexanes going to 80% hexanes / 20% EtOAc to give 48 g (62.1 mmol, 65.3% yield) of the title compound as a pale yellow oil.1H-NMR (400MHz, MeOD, d): 7.36-7.29 (m, 5H), 5.11 (s, 2H), 4.10 (t, J= 6.8 Hz, 2H), 3.05 (t, J= 6.4 Hz, 2H), 2.44 (m, 6H), 2.37 (m, 2H), 2.30 (m, 2H), 1.65- 1.55 (m, 6H), 1.46 (m, 7H), 1.43 (m, 9H), 1.33 (m, 12H), 1.29 (m, 22H), 0.91 (t, J= 6.8 Hz, 6H); carbamate proton not observed.Step 4: General procedure for Boc cleavage - preparation of 3-hexylnonyl 8-[4-aminobutyl-(8-benzyloxy-8-oxo-octyl)amino]octanoate, HCI salt (Intermediate XXIII)o... li r 'Ofin HCI 4M in EtOAc RTC42H7SN2O4. HCI MW: 673.08 (709.54)XXII XXIII

[0332] A mixture of 3-hexylnonyl 8-[(8-benzyloxy-8-oxo-octyl)-[4-(tert-butoxycarbonylamino) butyl]amino]octanoate (Intermediate XXII; 32 g, 41.4 mmol, 1 eq) in HCl / EtOAc (4 M, 160.0 mL, 15.5 eq) was degassed and purged with N2 three times, then stirred at 20 °C for 1 h under dry N2. The mixture was concentrated in vacuo to give 34 g (crude, HC1- 109 - LEGAL\113833648\6salt) of the title compound as a red oil, which was carried through to the next step without further purification. 'H-NMR (400MHz, MeOD, d): 7.36-7.28 (m, 5H), 5.11 (s, 2H), 4.10 (t, J= 6.8 Hz, 2H), 3.20-3.12 (m, 6H), 3.01 (t, J= 7.6 Hz, 2H), 2.37 (m, 2H), 2.32 (m, 2H), 1.90-1.82 (m, 2H), 1.74 (m, 6H), 1.66-1.56 (m, 6H), 1.45-1.36 (m, 12H), 1.29 (m, 21H), 0.91 (t, J = 6.6 Hz, 6H); amine salt protons not observed.Step 5: General procedure for headgroup amide coupling - Preparation of 3-hexylnonyl 8-[4-[3- (benzenesulfonyl)propanoylamino]butyl-(8-benzyloxy-8-oxo-octyl)amino]octanoate (Intermediate XXIV; lipid 141)0 Li EDC-HOS DMAP< o El3N..... N A HOL HjNx xO DCM XXti! C51 H84N2O7S MW: 869.30

[0333] To a solution of 3 -hexylnonyl 8-[4-aminobutyl-(8-benzyloxy-8-oxo- octyl)amino]octanoate (Intermediate XXIII; 16 g, 22.6 mmol, 1 eq. as HC1 salt) and 3- (benzenesulfonyl)propanoic acid (4.83 g, 22.6 mmol, 1 eq) in DCM (160 mL) was added EDC- HC1 (8.65 g, 45.1 mmol, 2 eq), triethylamine (EtsN; 6.85 g, 67.7 mmol, 9.42 mL, 3 eq) and DMAP (275 mg, 2.26 mmol, 0.1 eq) and the reaction stirred at 20 °C for 12 h under a dry N2 atmosphere. The mixture was concentrated in vacuo and the residue purified by silica gel chromatography, eluting with 100% hexanes going to 100% EtOAc to give 16 g (18.4 mmol, 81.6% yield) of the title compound as a pale yellow oil. UPLC-MS: m / z (ESI) for C51H84N2O7S [M+H]+expected:869.61; observed: 869.7; 'H-NMR (400MHz, CDCh, d): 7.91 (m, 2H), 7.65 (m, 1H), 7.57 (m, 2H), 7.37 (m, 5H), 6.77 (br s, 1H), 5.12 (s, 2H), 4.09 (t, J= 7.2 Hz, 2H), 3.46 (m, 2H), 3.20 (m, 2H), 2.62 (m, 2H), 2.37 (m, 8H), 2.29 (m, 2H), 1.68-1.56 (m, 6H), 1.55-1.51 (m, 4H), 1.41 (m, 4H), 1.35-1.24 (m, 33H), 0.89 (t, J= 6.8 Hz, 6H).- 110 - LEGAL\113833648\6B-9. Monoamine synthesis - Scheme 7: Synthesis of benzyl 8-((8-((3-hexylnonyl)oxy)-8- oxooctyl)(4-(3 -(phenylsulfonyl)propanamido)butyl)amino)octanoate (Intermediate XXVII) Scheme 7.intermediate IIIKI O K2CO3K acetonitrileOBn80 °CHCI4M in EtOAcRT XXVEDC-HClDMAPEt3NDCMStep 1: Preparation of benzyl 8-[4-(tert-butoxycarbonylamino)butyl-[8-(l-butylnonoxy)-8-oxo- octyl]amino]octanoate (Intermediate XXV)Intermediate IIIKI K2CO3acetonitrile 80 °CC45H80N2O5MW: 745.14XXV

[0334] Following the general procedure for second alkylation of core amine benzyl 8-[4-(tert- butoxycarbonylamino)butylamino]octanoate (Intermediate XXI; 35 g, 83.2 mmol, 1 eq) was reacted with 1-butylnonyl 8-bromooctanoate (Intermediate III; 42 g, 103.6 mmol, 1.24 eq) followed by an aqueous workup. The residue was purified by silica gel chromatography, eluting with 100% petroleum ether going to 100% EtOAc to give 48 g (64.4 mmol, 77.4% yield) of the title compound as a yellow oil. 'H-NMR. (400MHz, CDCh, d): 7.36-7.29 (m, 5H), 5.12 (s, 2H), 4.87 (m, 1H), 3.12 (br d, J= 5.6 Hz, 2H), 2.43-2.31 (m, 7H), 2.30-2.24 (m, 2H), 1.68-1.56 (m, - Ill - LEGAL\113833648165H), 1.55-1.36 (m, 16H), 1.35-1.21 (m, 34H), 0.89 (m, 6H)Step 2: Preparation of benzyl 8-[4-aminobutyl-[8-(l-butylnonoxy)-8-oxo-octyl]amino]octanoate (Intermediate XXVI)o OBn;■..., A OSn0HCI Jo£ 4M in EtOAc > 'cr ' HCI. HjN ■'Z- -Q'RTC40H72N2O4. HCl XXV MW: 645.03 (681.49)XXVI

[0335] A mixture of benzyl 8-[4-(tert-butoxycarbonylamino)butyl-[8-(l-butylnonoxy)-8-oxo- octyl]amino]octanoate (Intermediate XXV; 27 g, 36.2 mmol, 1 eq) in HCl / EtOAc (4 M, 200.00 mL, 22.1 eq) was degassed and purged with N2 three times, and the reaction was stirred at 25 °C for 1 h. The mixture was concentrated in vacuo to give 28 g (crude, HCI) of the title compound as a red oil which was carried through to the next step without further purification. 'H-NMR (400MHz, MeOD, d): 7.37-7.31 (m, 5H), 5.11 (s, 2H), 4.88 (m, 1H; partially obscured by water peak), 3.18-3.14 (m, 6H), 3.01 (m, 2H), 2.38 (t, J= 7.2 Hz, 2H), 2.32 (m, 2H), 1.87-1.81 (m, 2H), 1.78-1.70 (m, 6H), 1.68-1.61 (m, 4H), 1.57-1.52 (m, 4H), 1.49-1.35 (m, 12H), 1.34-1.23 (m, 16H), 0.91 (m, 6H); amine salt protons not observed.Step 3: Preparation of benzyl 8-[4-[3-(benzenesulfonyl)propanoylamino]butyl-[8-(l- butylnonoxy)-8-oxo-octyl]amino]octanoate (Intermediate XXVII; Compound 140)EDC-HCl DMAP Et3N HCI. H;. MDCMXXVI C49HS0N2O7S MW: 841.25XXVII

[0336] To a solution of benzyl 8-[4-aminobutyl-[8-(l-butylnonoxy)-8-oxo- octyl]amino]octanoate (Intermediate XXVI; 13 g, 19.1 mmol, 1 eq, HCI salt) and 3- (benzenesulfonyl)propanoic acid (4.09 g, 19.1 mmol, 1 eq) in DCM (130 mL) was added EDC- HCI (7.31 g, 38.1 mmol, 2 eq), EtsN (5.79 g, 57.2 mmol, 7.97 mL, 3 eq) and DMAP (233.0 mg, 1.91 mmol, 0.1 eq) ) and the reaction stirred at 20 °C for 12 h under a dry N2 atmosphere. The mixture was concentrated in vacuo and the residue purified by silica gel chromatography, eluting with 100% hexanes going to 10% hexanes / 90% EtOAc to give 13 g (15.4 mmol, 81.0% yield) of the title compound as a colorless oil. UPLC-MS: m / z (ESI) for C49H80N2O7S [M+H]+expected:- 112 - LEGAL\113833648\6841.58; observed: 841.7; 'H-NMR (400MHz, CDCh, d): 7.91 (m, 2H), 7.65 (m, 1H), 7.58 (m, 2H), 7.38 (m, 5H), 6.76 (br s, 1H), 5.12 (s, 2H), 4.88 (m, 1H), 3.46 (t, J = 7.8 Hz, 2H), 3.18 (m, 2H), 2.61 (t, J= 7.8 Hz, 2H), 2.48-2.33 (m, 8H), 2.29 (m, 2H), 1.70-1.58 (m, 4H), 1.56-1.50 (m, 8H), 1.41 (m, 4H), 1.36-1.20 (m, 28H), 0.90 (t, J= 6.8 Hz, 6H).B-10. Monoamine synthesis - Scheme 8a: Synthesis of 8-[4-( / -butoxycarbonylamino) butyl-[8- (3-hexylnonoxy)-8-oxo-octyl]amino]octanoic acid (Intermediate XXVIII)Scheme 8a.H2Pd(OH)2 / Al2O3THF (flow reactor) XXII C40H78N2O5MW: 683.07 XXVIII

[0337] Following the general procedure for benzyl ester hydrogenolysis by flow, 3 -hexylnonyl 8-[(8-benzyloxy-8-oxo-octyl)-[4-( / c / 7-butoxycarbonylamino)butyl]amino]octanoate (Intermediate XXII; 16 g, 20.7 mmol, 1 eq) in THF (320 mL) was hydrogenated in a flow reactor, the product collected from the reactor output and concentrated in vacuo to give 13 g (19.0 mmol, 92.0% yield) of the title compound as a colorless oil, which was carried through without further purification. 'H-NMR (400MHz, MeOD, d): 4.10 (t, J= 6.8 Hz, 2H), 3.18-3.08 (m, 8H), 2.32 (t, J = 7.2 Hz, 2H), 2.17 (m, 2H), 1.74- 1.52 (br. m, 14H), 1.44 (m, 9H), 1.39 (m, 12H), 1.30 (m, 22H), 0.89 (m, 6H), carbamate proton not observed.B-l 1. Monoamine synthesis - Scheme 8b: General procedure for benzyl ester hydrogenolysis over palladium on carbon - synthesis of 8-[4-[3-(benzenesulfonyl)propanoylamino]butyl-[8-(l- butylnonoxy)-8-oxo-octyl]amino]octanoic acid, HC1 salt (Intermediate XXIX)Scheme 8b.H210% Pd / C THFC44H78N2O7S . HCl MW: 779.18 (815.64)XXIX

[0338] To a solution of benzyl 8-[4-[3-(benzenesulfonyl)propanoylamino]butyl-[8-(l- butylnonoxy)-8-oxo-octyl]amino]octanoate (Intermediate XXIV; 11.7 g, 13.9 mmol, 1 eq) in THF (35 mL) was added 10% Pd / C (5.92 g, 5.56 mmol, 0.4 eq) with stirring under dry N2 to give a suspension. The reaction vessel was evacuated and back-filled with H2 several times, and the reaction stirred under H2 (40 psi) at 20 °C for 16 hours. The reaction vessel was purged with nitrogen, the mixture filtered through diatomaceous earth and the filtrate concentrated in vacuo.- 113 - LEGAL\113833648\6The residue was purified by silica gel chromatography, eluting with 100% DCM going to 50%DCM / 50% MeOH, then further purified by prep-HPLC (column: FLM Titank Cl 8 Bulk 250x70mm lOu; mobile phase: [H2O (0.05% HCl)-MeCN: THF=l:l]; gradient: 40%-80% B over 20.0 min) to give 5.5 g (6.98 mmol, 52.5% yield, HC1) of the title compound as a colorless oil. Compound is used as is or may be converted to the free base (partition between DCM and a saturated aqueous NaHCO3solution) prior to use. UPLC-MS: m / z (ESI) for C44H78N2O7S [M+H]+expected: 779.56; observed: 779.7; 'H-NMR (400MHz, DMSO-d6, d): 11.99 (br s, 1H), 9.87 (br s, 1H), 8.09 (m, 1H), 7.89 (m, 2H), 7.75 (m, 1H), 7.67 (m, 2H), 4.02 (t, J= 6.8 Hz, 2H), 3.59 (t, J = 7.6 Hz, 2H), 3.02-2.96 (m, 8H), 2.42 (m, 2H), 2.26 (m, 2H), 2.19 (m, 2H), 1.60 (m, 6H), 1.49 (m, 6H), 1.26 (m, 4H), 1.22 (m, 32H), 0.85 (m, 6H).B-12. Monoamine synthesis - Scheme 8c: Synthesis of 8-[4-( / -butoxycarbonylamino) butyl-[8- (l-butylnonoxy)-8-oxo-octyl]amino]octanoic acid (Intermediate XXX)Scheme 8c.o H, <... It OH Pd(OH)2 / Al2O3THF (flow reactor) MW: 655.02

[0339] Following the general procedure for benzyl ester hydrogenolysis by flow, benzyl 8-[4- (tert-butoxycarbonylamino)butyl-[8-(l-butylnonoxy)-8-oxo-octyl]amino]octanoate (Intermediate XXV; 16 g, 21.5 mmol, 1 eq) in THF (320 mL) was hydrogenated in a flow reactor, the product collected from the reactor output and concentrated in vacuo to give 13 g (19.8 mmol, 92.4% yield) of the title compound as a white solid that was carried through without further purification. 'H-NMR (400MHz, CDCh, d): 4.85 (m, 1H), 3.14 (br s, 2H), 2.73-2.69 (m, 6H), 2.30-2.24 (m, 4H), 1.61 (m, 6H), 1.51 (m, 10H), 1.44 (m, 9H), 1.32-1.15 (m, 28H), 0.88 (m, 6H), carboxylic acid and carbamate protons not observed.B-13. Monoamine synthesis - Scheme 8d: Synthesis of 8-[4-[3-(benzenesulfonyl) propanoylamino]butyl-[8-(l-butylnonoxy)-8-oxo-octyl]amino]octanoic acid, HCI salt (Intermediate XXXI)Scheme 8d.XXVII C42H74N2O7S. HCl MW: 751.12 (787.56)XXX!- 114 - LEGAL\113833648\6

[0340] Following the general procedure for benzyl ester hydrogenolysis over palladium on carbon, benzyl 8-[4-[3-(benzenesulfonyl)propanoylamino]butyl-[8-(l-butylnonoxy)-8-oxo-octyl]amino]octanoate (Intermediate XXVII; 11.7 g, 13.9 mmol, 1 eq) in THF (35 mL) was hydrogenated, the product purified by silica gel chromatography (100% DCM going to 50% DCM / 50% MeOH) followed by prep HPLC (column: FLM Titank C18 Bulk 250x70mm lOu; mobile phase: [H20(0.05% HCl)-MeCN: THF=l:l];gradient:40%-80% B over 20.0 min) to give 5.5 g (6.98 mmol, 52.4% yield, HC1) of the title compound as a colorless oil. Compound is used as is or may be converted to the free base (partition between DCM and a saturated aqueous NaHCO3solution) prior to use. UPLC-MS: m / z (ESI) for C44H79N2O5[M+H]+expected: 751.53; observed: 751.7; 'H-NMR (400MHz, DMSO-d6d): 11.99 (br s, 1H), 9.56 (br s, 1H), 8.06 (m, 1H), 7.89 (m, 2H), 7.75 (m, 1H), 7.67 (m, 2H), 4.78 (m, 1H), 3.50 (t, J= 7.4 Hz, 2H), 3.02-2.97 (m, 8H), 2.43 (m, 2H), 2.19 (m, 4H), 1.59 (m, 6H), 1.49 (m, 8H), 1.26 (m, 2H), 1.23 (m, 28H), 0.85 (m, 6H).B-14. Monoamine synthesis - Scheme 9: Synthesis of 3 -hexylnonyl 8-[[8-(l-butylnonoxy)-8-oxo-octyl]amino]octanoate, HC1 salt (Intermediate XXXIII)Scheme 9.EDC-HClDMAPEt3NHO DCMXXiHXXXIII- 115 - LEGAL\113833648\6Step 1: Preparation of 3-hexylnonyl 8-[(8-benzyloxy-8-oxo-octyl)-[4-(3- phenoxypropanoylamino) butyl]amino]octanoate (Intermediate XXXII; Compound 142)HOEDC-HCl DMAPEt3N DCMC51H84N2O6XXIIIMW: 821.24XXXI I

[0341] Following the general procedure for headgroup amide coupling, 3-hexylnonyl 8-[4- aminobutyl-(8-benzyloxy-8-oxo-octyl)amino]octanoate (Intermediate XXIII; 16 g, 22.6 mmol, 1 eq, HC1) was reacted with 3-phenoxypropanoic acid (3.75 g, 22.6 mmol, 1 eq) followed by an aqueous workup. Purification by silica gel chromatography eluting with 100% hexanes going to 100% EtOAc gave 14 g (17.0 mmol, 75.6% yield) of the title compound as a light yellow oil.UPLC-MS: m / z (ESI) for C51H84N2O6[M+H]+expected: 821.64; observed: 821.8; 'H-NMR (400MHz, MeOD, d): 7.38-7.23 (m, 7H), 6.92 (m, 3H), 5.11 (s, 2H), 4.23 (t, J= 6.0 Hz, 2H), 4.10 (t, J= 6.8 Hz, 2H), 3.26 (m, 2H; partially obscured by water peak), 2.90-2.71 (m, 6H), 2.63 (t, J= 6.0Hz, 2H), 2.35 (t, J= 7.2 Hz, 2H), 2.30 (t, J = 7.4 Hz, 2H), 1.68-1.52 (m, 14H), 1.35- 1.27 (m, 33H), 0.90 (t, J= 6.8 Hz, 6H)Step 2: Preparation of 8-[[8-(3-hexylnonoxy)-8-oxo-octyl]-[4-(3- phenoxypropanoylamino)butyl]amino]octanoic acid, HC1 salt (Intermediate XXXIII)o H210% Pd / C JI THF XXXII C44H78N2O5. HCl MW: 731.12 (767.58)XXXIII

[0342] Following the general procedure for benzyl ester hydrogenolysis over palladium on carbon, 3-hexylnonyl 8-[(8-benzyloxy-8-oxo-octyl)-[4-(3-phenoxypropanoylamino)butyl]amino]- octanoate (Intermediate XXXII; 11 g, 13.39 mmol, 1 eq) in THF (220 mL) was hydrogenated under H2 (20 psi) at 20 °C for 16 hours. The mixture was fdtered and the filtrate was concentrated in vacuo to give a yellow oil. This was purified by prep-HPLC (column: WePure Biotech Phenyl- Hexyl 250x707u; mobile phase: [H20(0.05% HCl)-MeCN: THF=l:l]; gradient: 40%-70% B over 20.0 min) to give 6.8 g (9.30 mmol, 69.4% yield, HC1) of the title compound as a colorless oil.Compound is used as is or may be converted to the free base (partition between DCM and a saturated aqueous NaHCO3solution) prior to use. UPLC-MS: m / z (ESI) for C44H78N2O5[M+H]+expected: 731.59; observed: 731.7. 'H-NMR (400MHz, DMSO-d6d): 11.98 (br s, 1H), 9.59 (br- 116 - LEGAL\113833648\6s, 1H), 8.06 (t, J= 5.8 Hz, 1H), 7.89 (m, 2H), 7.76 (m, 1H), 7.67 (t, J= 7.6 Hz, 2H), 4.03 (t, J = 6.8 Hz, 2H), 3.50 (m, 2H), 3.03-2.98 (m, 8H), 2.42 (m, 2H), 2.26 (m, 2H), 2.18 (m, 2H), 1.59 (m, 6H), 1.50 (m, 6H), 1.26 (m, 3H), 1.22 (m, 32H), 0.85 (t, J= 6.8, 6H).B-15. Monoamine synthesis - Scheme 10: Synthesis of 8-[4-[4-(4-bromophenyl) butanoyl amino]butyl-[8-(l-butylnonoxy)-8-oxo-octyl]amino] octanoic acid (Intermediate XXXVI) Scheme 10.HO4M in EtOAcRTXXVIIIIntermediate XXXVDMAP DIEA2 HCI HjN ''DCM XXXIVXXXVIStep 1: Preparation of 8-[4-aminobutyl-[8-(3-hexylnonoxy)-8-oxo-octyl]amino]octanoic acid (Intermediate XXXIV)o o, J, A <- • OH s' ' ' OH J HCI J9 f 9 7 ‘ ' 4M in EtOAc s' 9 f N..-x..x w... A.x 1 O’ N’ ' 'O' - 'O'H RT XXVIII C35H70N2O4· 2HClMW: 582.95 (655.87)XXXIV

[0343] Following the general procedure for Boc cleavage, 8-[4-(tert- butoxycarbonylamino)butyl-[8-(3-hexylnonoxy)-8-oxo-octyl]amino]octanoic acid (Intermediate XXVIII; 13 g, 19.0 mmol, 1 eq) was deprotected with HCl / EtOAc (4 M, 95.2 mL, 20 eq) and the- 117 - LEGAL\113833648\6mixture concentrated in vacuum to give 12.5 g (crude, HC1) of the title compound as a yellow oil which was carried through without further purification. 'H-NMR (400MHZ, MeOD, d): 4.09 (t, J = 6.8 Hz, 2H), 3.18-3.12 (m, 6H), 3.01 (m, 2H), 2.31 (m, 4H), 1.88 (m, 2H), 1.75 (m, 6H), 1.64- 1.55 (m, 6H), 1.48-1.35 (m, 12H), 1.33-1.21 (m, 21H), 0.91 (t, J= 6.8 Hz, 6H); amine salt protons not observed.Step 2: Preparation of (2,5-dioxopyrrolidin-l-yl) 4-(4-bromophenyl) butanoate (Intermediate XXXV)C14H14BrNO4MW: 340.17

[0344] To a solution of 4-(4-bromophenyl)butanoic acid (20 g, 82.3 mmol, 1 eq) and N- hydroxysuccinimide (10.42 g, 90.5 mmol, 1.1 eq) in EtOAc (200 mL) was added DCC (18.7 g, 90.5 mmol, 18.3 mL, 1.1 eq) in portions with stirring under dry N2 and the reaction stirred at 25 °C for 16 h. The mixture was filtered, the filtrate diluted with water (500 mL), the organic phase separated and the aqueous phase extracted with ethyl acetate (2x 200 mL). The combined organic phase was washed with brine (2x 100 mL), dried over anhydrous Na2SC>4, filtered and filtrate concentrated in vacuo to give 25 g, (crude) of the title compound as a light yellow solid which was used without further purification. 'H-NMR (400MHz, MeOD, d): 7.41 (d, J= 8.4 Hz, 2H), 7.15 (d, J= 8.4 Hz, 2H), 2.70 (s, 4H), 2.69 (t, J = Hz, 2H), 1.35-1.24 (m, 2H), 0.89 (t, J= 6.8 Hz, 2H).Step 3: Preparation of 8-[4-[4-(4-bromophenyl) butanoylamino] butyl-[8-(l-butylnonoxy)-8-oxo- octyl]amino] octanoic acid (Intermediate XXXVI)intermediate XXXV DMAP DIEA DCM XXXiVMW: 808.04XXXVI

[0345] To a mixture of 8-[4-aminobutyl-[8-(l-butylnonoxy)-8-oxo-octyl]amino]octanoic acid (Intermediate XXXIV; 9.5 g, 16.1 mmol, 1 eq, HC1) and (2,5-dioxopyrrolidin-l-yl) 4-(4- bromophenyl)butanoate (Intermediate XXXV; 5.46 g, 16.1 mmol, 1 eq) in DCM (95 mL) was added DMAP (196.27 mg, 1.61 mmol, 0.1 eq) and DIEA (6.23 g, 48.2 mmol, 8.39 mL, 3 eq) in one portion at 20 °C under N2 atmosphere The mixture was stirred at 20 °C for 2 hours. The- 118 - LEGAL\113833648\6mixture was concentrated in vacuum to get a residue. The residue was purified by silica gel chromatography (SiCL, Dichloromethane: Methanol=l / 0, 9 / 1) to give 3.8 g (4.87 mmol, 30.3% yield) of the title compound as a yellow oil. UPLC-MS: m / z (ESI) for C45H79BrN2O7[M+H]+expected:807.52 / 809.52; observed: 807.5 / 809.5; 'H-NMR (400MHz, DMSO-d6, d): 9.75 (br s, 1H), 7.86 (t, J= 5.6 Hz, 1H), 7.44 (d, J= 8.4 Hz, 2H), 7.15 (d, J = 8.4 Hz, 2H), 4.04 (t, J= 6.8 Hz, 2H), 2.96-3.04 (m, 8H), 2.51 (m, 2H; partially obscured by solvent peak), 2.29-2.21 (m, 4H), 2.06 (m, 2H), 1.91-1.86 (m, 2H), 1.71-1.54 (m, 6H), 1.52-1.36 (m, 9H), 1.35-1.27 (m, 32H), 0.87 (m, 6H).B-16. Synthesis of / V-(4-aminobutyl)-3-(phenylsulfonyl)propenamide, HCI salt (Intermediate LI)DCC DIEA DCM0 °C - RTC19H29N2O5SMW: 384.49HCI / dioxaneC13H20N2O3S (HCl)MW: 284.37 (320.83)

[0346] To a solution of 3-(benzenesulfonyl)propanoic acid (10 g, 46.7 mmol, 1 equiv.) in DCM at 0 °C was added DCC (12 g, 58.3 mmol, 1.25 equiv.) with stirring, and after 5 minutes N-Boc-l,4-diaminobutane (9.7 g, 51.3 mmol, 1.1 equiv.) was added. The resulting mixture was stirred overnight at room temperature, fdtered, and the filtrate washed with 0.5M HCI, a saturated NaHCO3solution, and brine, dried over anhydrous MgSO4, filtered, and the filtrate cone, in vacuo. Purification of the residue with silica gel chromatography, eluting with 0%-10% MeOH in DCM provided 15 g (39 mmol) of the Boc-protected intermediate L. This was dissolved in a 4M solution of HCI in 1,4-dioxane and the mixture stirred at room temperature for two hours. The reaction mixture was then concentrated in vacuo to give 12.5 g (39 mmol, 83.5% yield over two steps) of the title product as an HCI salt. UPLC-MS: m / z (ESI) for C13H20N2O3S [M+H]+ expected: 285.37; observed: 285.22; 'H NMR (400 MHz, MeOD) 87.64 - 7.55 (m, 2H), 7.45 -7.36 (m, 1H), 7.35 - 7.26 (m, 2H), 3.18 (t, J= 7.3 Hz, 2H), 2.82 (t, J = 6.7 Hz, 2H), 2.59 (t, J = 7.5 Hz, 2H), 2.25 (t, J = 7.4 Hz, 2H), 1.39 - 1.15 (m, 4H).- 119 - LEGAL\113833648\6B-17. General procedure for Boc cleavage followed by amide formation - preparation of 3-hexylnonyl 8-((8-oxo-8-(tridecan-5-yloxy)octyl)(4-(3-(phenylthio)propanamido)butyl)amino) octanoate (Compound 1)V1. HCI / dioxaneDCM o2. EDC-HC! J 1..DMAP fDIEA. <0- o;.k jj;DCM '*-■ S' 'N- -O'C57H104N2O5SMW: 929.53Compound 1

[0347] To solution of 3 -hexylnonyl 8-[4-(tert-butoxycarbonylamino)butyl-[8-(l-butylnonoxy)-8-oxo-octyl]amino]octanoate (Intermediate V; 165 mg, 0.19 mmol, 1 eq) in 5 ml DCM was added 0.5 mL (4M in 1,4-di oxane; excess) and the solution stirred at RT for 1.5 hours, concentrated, and the residue redissolved in DCM. To the resulting solution were added 3-(phenylthio)propanoic acid (42 mg, 0.23 mmol, 1.2 eq), EDC-HC1 (44 mg, 0.23 mmol, 1.2 eq), and DIEA (0.2 mL, 1.14 mmol, 6 eq) and the reaction stirred overnight at room temp. The resulting mixture was diluted with DCM, washed with saturated aqueous sodium bicarbonate solution, the organics dried over Na2SO4, filtered, and the filtrate concentrated in vacuo. The residue was purified by silica gel chromatography, eluting with 100% DCM going to 87% DCM / 12% MeOH / 1% NH4OH to give 54.5 mg (0.059 mmol, 31.0% yield) of the title compound. UPLC-MS: m / z (ESI) for C57H104N2O5S: [M+H]+expected: 931.54; observed: 932.11. 'HNMR (400 MHz, CDCh) 87.32 (dd, J= 22.2, 7.4 Hz, 4H), 7.25 - 7.12 (m, 1H), 6.38 (s, 1H), 4.95 -4.76 (m, 1H), 4.07 (t, J= 7.1 Hz, 2H), 3.22 (dd, J= 9.1, 6.2 Hz, 4H), 2.46 (d, J= 7.4 Hz, 2H), 2.44 - 2.30 (m, 7H), 2.27 (t, J = 7.5 Hz, 4H), 1.55 (ddt, J= 28.6, 12.7, 6.8 Hz, 17H), 1.27 (d, J = 14.7 Hz, 50H), 0.88 (td, J= 6.9, 2.9 Hz, 12H).

[0348] Following the general procedure for Boc cleavage followed by amide formation, the following compounds were prepared from 3-hexylnonyl 8-[4-( / c / 7-butoxy carbonyl amino)butyl-[8-(l-butylnonoxy)-8-oxo-octyl]amino]octanoate (Intermediate V) and the corresponding amines.- 120 - LEGAL\113833648\6CggH-i 06^20?MW: 943.49Compound 2

[0349] 3-hexylnonyl 8-((4-(3-(4-methoxyphenoxy)propanamido)butyl)(8-oxo-8-(tridecan-5-yloxy)octyl)amino)octanoate (Compound 2). UPLC-MS: m / z (ESI) for C58H106N2O7: [M+H]+expected: 944.49; observed: 946.26. ' H NMR (400 MHz, CDCh) 57.09 (s, 1H), 6.92 - 6.75 (m, 4H), 4.84 (p, J = 6.3 Hz, 1H), 4.20 (t, J= 6.1 Hz, 2H), 4.06 (t, J= 7.1 Hz, 2H), 3.75 (d, J = 7.5 Hz, 4H), 3.69 - 3.59 (m, 2H), 3.33 (q, J= 6.2 Hz, 2H), 3.08 - 2.80 (m, 6H), 2.67 (t, J= 6.1 Hz, 2H), 2.36 - 2.21 (m, 4H), 1.96 - 1.80 (m, 1H), 1.73 (dq, J= 12.0, 6.3 Hz, 2H), 1.69 - 1.45 (m, 11H), 1.42 - 1.15 (m, 50H), 0.87 (tt, J= 7.1, 1.8 Hz, 12H).C57H1Q4N2O7SMW: 961.53Compound 4

[0350] 3-hexylnonyl 8-((8-oxo-8-(tridecan-5-yloxy)octyl)(4-(3-(phenylsulfonyl)propanamido) butyl)amino)octanoate (Compound 4). UPLC-MS: m / z (ESI) for C57H104N2O7S: [M+H]+expected: 962.53; observed: 962.22. 'H NMR (400 MHz, CDCh) 58.00 - 7.88 (m, 2H), 7.73 - 7.63 (m, 1H), 7.57 (t, J= 7.7 Hz, 2H), 6.76 (s, 1H), 4.86 (s, 1H), 4.08 (t, J= 7.1 Hz, 2H), 3.58 - 3.37 (m, 2H), 3.18 (q, J = 6.1 Hz, 2H), 2.69 - 2.55 (m, 2H), 2.40 (s, 6H), 2.28 (t, J= 7.5 Hz, 4H), 1.72 - 1.55 (m, 6H), 1.51 (dt, J = 9.3, 6.1 Hz, 5H), 1.41 (d, J= 7.5 Hz, 6H), 1.37- 1.17 (m, 50H), 0.88 (td, J=6.1, 3.6 Hz, 12H).CsaH 105^3^7MW.' 956.49Compound 5

[0351] 3-hexylnonyl 8-((4-(3-(4-carbamoylphenoxy)propanamido)butyl)(8-oxo-8-(tridecan-5-yloxy)octyl)amino)octanoate (Compound 5). UPLC-MS: m / z (ESI) for C58H105N3O7: [M+H]+expected: 957.49; observed: 957.32. 'H NMR (500 MHz, CDCh) 87.86 - 7.71 (m, 2H), 7.02 - 6.88 (m, 2H), 6.83 (s, 1H), 4.97 - 4.71 (m, 1H), 4.31 (t, J= 6.1 Hz, 2H), 4.07 (t, J= 7.1 Hz, 2H), 3.29 (q, J= 6.0 Hz, 2H), 2.66 (t, J= 6.2 Hz, 2H), 2.27 (t, J= 7.5 Hz, 4H), 1.81 - 1.54 (m, 8H),- 121 - LEGAL\113833648\61.54 - 1.46 (m, 8H), 1.44 - 1.13 (m, 50H), 0.87 (td, J= 6.9, 3.2 Hz, 12H).MW: 992.36Compound 64

[0352] 3-hexylnonyl 8-((4-(3-(4-bromophenoxy)propanamido)butyl)(8-oxo-8-(tridecan-5-yloxy)octyl)amino)octanoate (Compound 64). UPLC-MS: m / z (ESI) for CsrHiosBr^Ch: [M+H]+expected 993.36; observed: 993.03.XH NMR (500 MHz, CDC13) 57.45 - 7.31 (m, 2H), 6.87-6.73 (m, 2H), 6.55 (s, 1H), 4.86 (t, J= 6.3 Hz, 1H), 4.23 (t, J= 6.2 Hz, 2H), 4.07 (t, J= 7.1 Hz, 2H), 3.85 - 3.60 (m, 2H), 3.27 (q, J= 6.3 Hz, 2H), 2.61 (t, J= 6.2 Hz, 2H), 2.39 (d, J= 12.5 Hz, 5H), 2.27 (t, J = 7.5 Hz, 4H), 1.70 - 1.46 (m, 12H), 1.42 (dt, J= 15.3, 5.5 Hz, 5H), 1.35 - 1.13 (m, 49H), 0.88 (td, J= 7.0, 3.0 Hz, 12H).C58HWN3OSMW: 926.51Compound 7

[0353] 3-hexylnonyl 8-((4-(3-(methyl(phenyl)amino)propanamido)butyl)(8-oxo-8-(tridecan-5-yloxy)octyl)amino)octanoate (Compound 7). UPLC-MS: m / z (ESI) for C58H107N3O5: [M+H]+expected: 927.51; observed: 928.16. 'H NMR (400 MHz, CDCh) 57.26 (s, 3H), 5.77 - 4.95 (m, 9H), 4.49 (s, 2H), 4.02 (q, J= 10.4 Hz, 3H), 3.22 (s, 4H), 2.49 (d, J = 42.9 Hz, 11H), 2.01 (q, J = 6.9 Hz, 14H), 1.83 (s, 4H), 1.48 (s, 8H), 1.40 - 0.99 (m, 29H), 0.89 (t, J= 6.7 Hz, 12H).C57H1Q3CIN2O5MW: 947.91Compound 8

[0354] 3-hexylnonyl 8-((4-(3-(4-chlorophenoxy)propanamido)butyl)(8-oxo-8-(tridecan-5-yloxy)octyl)amino)octanoate (Compound 8). UPLC-MS: m / z (ESI) for C57H103CIN2O6: [M+H]+expected: 948.91; observed: 950.46. 'H NMR (400 MHz, CDCh) 57.22 (d, J= 8.5 Hz, 2H), 6.83 (d, J= 8.4 Hz, 2H), 4.86 (s, 1H), 4.23 (t, J= 6.2 Hz, 2H), 4.07 (t, J= 7.1 Hz, 2H), 3.28 (d, J= 6.1 Hz, 2H), 2.62 (t, J= 6.2 Hz, 2H), 2.39 (s, 6H), 2.27 (t, J= 7.6 Hz, 4H), 1.55 (ddt, J= 28.9, 14.1, 6.8 Hz, 10H), 1.48 - 1.39 (m, 4H), 1.28 (d, J= 20.7 Hz, 53H), 0.95 - 0.72 (m, 12H).- 122 - LEGAL\113833648\6*^58^1 ogNjOgMW: 927.49Compound 9

[0355] 3-hexylnonyl 8-((8-oxo-8-(tridecan-5-yloxy)octyl)(4-(3-(p-tolyloxy)propanamido) butyl)amino)octanoate (Compound 9). UPLC-MS: m / z (ESI) for C58H106N2O6: [M+H]+expected: 928.49; observed: 929.61. 'H NMR (400 MHz, CDCh) 87.07 (d, J= 8.0 Hz, 2H), 6.80 (d, J= 8.0 Hz, 2H), 6.51 (s, 1H), 4.86 (s, OH), 4.22 (t, J= 6.0 Hz, 2H), 4.07 (t, J= 7.1 Hz, 2H), 3.27 (q, J= 6.2 Hz, 2H), 2.62 (t, J= 6.1 Hz, 2H), 2.39 (s, 6H), 2.27 (q, J= 5.8 Hz, 7H), 1.57 (ddt,.7=29.1, 15.2, 6.6 Hz, 14H), 1.47- 1.33 (m, 4H), 1.27 (d, J= 14.3 Hz, 53H), 0.88 (q, J= 4.4 Hz, 12H).C57H104N2O6MW: 913.47Compound 10

[0356] 3-hexylnonyl 8-((8-oxo-8-(tridecan-5-yloxy)octyl)(4-(3-phenoxypropanamido)butyl) amino)octanoate (Compound 10). UPLC-MS: m / z (ESI) for C57H104N2O6: [M+H]+expected: 914.47; observed: 915.60. 'H NMR (400 MHz, CDCh) 87.26 (s, 2H), 7.04 - 6.86 (m, 3H), 4.26 (t, J= 6.1 Hz, 2H), 4.07 (t, J= 7.1 Hz, 2H), 3.29 (d, J= 6.8 Hz, 2H), 2.65 (t, J= 6.0 Hz, 2H), 2.41 (s, 6H), 2.35 -2.20 (m, 4H), 1.77- 1.35 (m, 22H), 1.28 (d, J= 19.7 Hz, 50H), 0.88 (td, J = 6.9, 2.6 Hz, 12H).CsfiHioghhOgMW: 911.49Compound 11

[0357] 3-hexylnonyl 8-((8-oxo-8-(tridecan-5-yloxy)octyl)(4-(4-phenylbutanamido)butyl) amino)octanoate (Compound 11). UPLC-MS: m / z (ESI) for C58H106N2O5: [M+H]+expected: 912.49; observed: 913.75. 'H NMR (400 MHz, CDCh) 87.36 - 7.23 (m, 2H), 7.23 - 7.14 (m, 3H), 6.08 (s, 1H), 4.94 - 4.76 (m, 1H), 4.07 (t, J= 7.1 Hz, 2H), 3.23 (q, J= 6.3 Hz, 2H), 2.65 (t, J = 7.5 Hz, 2H), 2.37 (dt, J= 14.9, 7.3 Hz, 6H), 2.27 (t, J= 7.5 Hz, 4H), 2.15 (t, J= 7.5 Hz, 2H), 2.04 - 1.91 (m, 2H), 1.74 - 1.45 (m, 16H), 1.39 (s, 1H), 1.28 (d,.7= 20.4 Hz, 50H), 0.88 (td, J =- 123 - LEGAL\113833648\66.9, 2.7 Hz, 12H).itC56H-102M2O7MW: 915.44Compound 12

[0358] 3-hexylnonyl 8-((4-(2-(3,4-dihydroxyphenyl)acetamido)butyl)(8-oxo-8-(tridecan-5-yloxy)octyl)amino)octanoate (Compound 12). UPLC-MS: m / z (ESI) for C56H102N2O7: [M+H]+expected: 916.44; observed: 917.55. 'H NMR (400 MHz, CDCh) 86.80 (dd, J= 7.9, 3.9 Hz, 1H), 6.73 - 6.52 (m, 2H), 5.67 (d, J= 25.4 Hz, 1H), 4.87 (t, J= 6.2 Hz, 1H), 4.08 (td, J= 7.1, 2.6 Hz, 2H), 3.46 (d, J= 3.2 Hz, 2H), 3.24 (d, J= 6.0 Hz, 2H), 2.47 (t, J= 8.2 Hz, 6H), 2.29 (t, J= 7.5 Hz, 4H), 1.71 - 1.36 (m, 14H), 1.36 - 1.11 (m, 55H), 0.88 (tt, J= 7.1, 1.8 Hz, 12H).oC58H105BrN2O5MW: 990.39Compound 13

[0359] 3-hexylnonyl 8-((4-(4-(4-bromophenyl)butanamido)butyl)(8-oxo-8-(tridecan-5-yloxy) octyl)amino)octanoate. (Compound 13). UPLC-MS: m / z (ESI) for CssHiosBr^CE: [M+H]+expected: 991.39; observed: 990.23. 'H NMR (400 MHz, CDCh) 57.47 - 7.26 (m, 2H), 7.18 -6.99 (m, 2H), 6.20 (s, 1H), 4.86 (t, J= 6.3 Hz, 1H), 4.07 (t, J= 7.1 Hz, 2H), 3.23 (q, J= 6.2 Hz, 2H), 2.60 (t, J= 7.5 Hz, 2H), 2.53 - 2.36 (m, 6H), 2.27 (t, J= 7.5 Hz, 4H), 2.13 (t, J= 7.4 Hz, 2H), 2.02 - 1.86 (m, 2H), 1.74 - 1.37 (m, 17H), 1.27 (d, J= 21.3 Hz, 50H), 0.87 (td, J= 6.9, 2.7 Hz, 12H).I N' JL 'N- V A -N- Avv,. A0A v.,v.H HC55H103N5O5MW: 914.46Compound 14

[0360] 3-hexylnonyl 8-((8-oxo-8-(tridecan-5-yloxy)octyl)(4-(3-(pyrimidin-2-ylamino) propanamido)butyl)amino)octanoate (Compound 14). UPLC-MS: m / z (ESI) for C55H103N5O5: [M+H]+expected: 915.46; observed: 915.20.XH NMR (400 MHz, CDCh) 58.26 (dt, J= 4.8, 1.4 Hz, 2H), 6.51 (s, 1H), 5.61 (s, 1H), 4.86 (p, J= 6.1 Hz, 1H), 4.07 (t, J= 7.0 Hz, 2H), 3.73 (q, J = 6.3 Hz, 2H), 3.23 (q, J= 6.2 Hz, 2H), 2.47 (t, J= 6.1 Hz, 2H), 2.37 (q, J= 7.6 Hz, 6H), 2.27 (t, J- 124 - LEGAL\113833648\6= 7.5 Hz, 4H), 1.69 - 1.43 (m, 14H), 1.39 (d, J= 6.7 Hz, 2H), 1.27 (d, J= 19.2 Hz, 52H), 0.87 (dq, J= 7.2, 3.7 Hz, 12H).^53^103^305MW: 922.48Compound 15

[0361] 3-hexylnonyl 8-((4-(2-(l / f-indol-3-yl)acetamido)butyl)(8-oxo-8-(tridecan-5-yloxy) octyl)amino)octanoate (Compound 15). UPLC-MS: m / z (ESI) for C58H103N3O5: [M+H]+expected: 923.48; observed: 924.55. 'H NMR (400 MHz, CDCh) 88.49 (s, 1H), 7.56 (d, J= 7.9 Hz, 1H), 7.40 (d, J = 8.1 Hz, 1H), 7.26 (s, 1H), 7.17- 7.08 (m, 2H), 5.87 (s, 1H), 4.87 (p, J = 6.2 Hz, 1H), 4.08 (t, J= 7.1 Hz, 2H), 3.19 (q, J= 6.5 Hz, 2H), 2.28 (t, J= 7.5 Hz, 8H), 1.73 - 1.45 (m, 16H), 1.27 (d, J= 16.3 Hz, 53H), 0.88 (td, J= 6.0, 3.3 Hz, 12H).C56H104H4O5MW: 913.47Compound 16

[0362] 3-hexylnonyl 8-((8-oxo-8-(tridecan-5-yloxy)octyl)(4-(3-(pyridin-2-ylamino) propanamido)butyl)amino)octanoate (Compound 16). UPLC-MS: m / z (ESI) for C56H104N4O5:[M+H]+expected: 914.47; observed: 914.00. 'H NMR (400 MHz, CDCh) 88.07 (d, J= 5.0 Hz, 1H), 7.37 (t, J= 7.9 Hz, 1H), 6.61 (s, 1H), 6.54 (t, J= 6.2 Hz, 1H), 6.40 (d, J= 8.4 Hz, 1H), 4.99 (s, 1H), 4.94 - 4.79 (m, 1H), 4.08 (t, J= 7.1 Hz, 2H), 3.67 (q, J= 6.2 Hz, 2H), 3.24 (q, J= 6.2 Hz, 2H), 2.48 (t, J= 6.2 Hz, 2H), 2.37 (s, 8H), 2.28 (t, J= 7.7 Hz, 4H), 1.81 - 1.55 (m,9H), 11.40 (m, 7H), 1.27 (d, J= 14.4 Hz, 48H), 0.87 (d, J= 6.9 Hz, 12H).MW: 991.55Compound 17

[0363] 3-hexylnonyl 8-((4-(3-(4-(methylsulfonyl)phenoxy)propanamido)butyl)(8-oxo-8- (tridecan-5-yloxy)octyl)amino)octanoate (Compound 17). UPLC-MS: m / z (ESI) for C58H106N2O8S: [M+H]+expected: 992.55; observed: 993.98. 'H NMR (400 MHz, CDCh) 87.86 (d, J= 8.6 Hz, 2H), 7.03 (d, J= 8.7 Hz, 2H), 4.85 (s, 2H), 4.36 (t, J= 6.6 Hz, 2H), 4.07 (t, J= 7.2- 125 - LEGAL\113833648\6Hz, 2H), 3.28 (s, 3H), 3.02 (s, 2H), 2.65 (s, 2H), 2.40 (s, 6H), 2.27 (t, J= 7.6 Hz, 4H), 1.55 (d, J = 13.0 Hz, 23H), 1.25 (s, 56H), 0.93 - 0.65 (m, 12H).CgrH-igjNjOsMW: 958.46Compound 18

[0364] 3-hexylnonyl 8-((4-(3-(4-nitrophenoxy)propanamido)butyl)(8-oxo-8-(tridecan-5-yloxy)octyl)amino)octanoate (Compound 18). UPLC-MS: m / z (ESI) for C57H103N3O8: [M+H]+expected: 959.46; observed: 961.22. 'H NMR (400 MHz, CDCh) 58.19 (d, J= 8.7 Hz, 2H), 6.96 (d, J= 8.7 Hz, 2H), 6.68 (s, 1H), 4.86 (s, 1H), 4.38 (t, J= 6.3 Hz, 2H), 4.07 (t, J= 7.1 Hz, 2H), 3.28 (t, J= 6.3 Hz, 2H), 2.66 (t, J= 6.3 Hz, 2H), 2.55 - 2.34 (m, 6H), 2.27 (t, J= 7.5 Hz, 4H), 1.55 (ddt, J= 28.8, 13.2, 6.6 Hz, 14H), 1.48 - 1.37 (m, 1H), 1.27 (d, J= 19.4 Hz, 52H), 0.87 (d, J = 7.3 Hz, 12H).o p y - I o C -■ CC■ k A,-xA - A XHCg / HiogNjOgSMW: 992.54Compound 19

[0365] 3-hexylnonyl 8-((8-oxo-8-(tridecan-5-yloxy)octyl)(4-(3-(4-sulfamoylphenoxy) propanamido)utyl)amino)octanoate (Compound 19). UPLC-MS: m / z (ESI) for C57H105N3O8S:[M+H]+expected: 993.54; observed: 994.23. 'H NMR (400 MHz, CDCh) 87.83 (d, J= 8.3 Hz, 2H), 6.94 (d, J= 8.3 Hz, 2H), 6.82 (d, J= 5.6 Hz, 1H), 4.85 (d, J= 6.3 Hz, 1H), 4.31 (t, J= 6.3 Hz, 2H), 4.07 (t, J= 7.1 Hz, 2H), 3.27 (q, J= 6.1 Hz, 2H), 2.63 (t, J= 6.3 Hz, 2H), 2.40 (q, J = 7.1 Hz, 6H), 2.26 (t, J = 7.5 Hz, 4H), 1.55 (ddt, J= 28.1, 13.5, 6.3 Hz, 12H), 1.41 (t, J= 7.5 Hz,6H), 1.27 (d, J= 15.8 Hz, 51H), 0.87 (d, J= 7.3 Hz, 12H).MW: 927.26Compound 20

[0366] (4-(2-((4-((8-((3-hexylnonyl)oxy)-8-oxooctyl)(8-oxo-8-(tridecan-5-yloxy)octyl)amino) butyl)amino)-2-oxoethyl)phenyl)boronic acid (Compound 20). UPLC-MS: m / z (ESI) for C56H103BN2O7: [M+H]+expected: 928.26; observed: 930.11. *HNMR (400 MHz, CDCh) 87.86- 126 - LEGAL\113833648\6(d, J= 71.9 Hz, 2H), 7.26 (s, 2H), 5.92 (d, J= 41.4 Hz, 1H), 4.86 (q, J= 6.4 Hz, 1H), 4.07 (t, J = 7.0 Hz, 2H), 3.56 (s, 1H), 3.28 - 2.92 (m, 2H), 2.62 - 2.03 (m, 8H), 1.87 - 1.46 (m, 7H), 1.27 (d, J= 15.9 Hz, 65H), 0.87 (d, J= 7.4 Hz, 12H).o oH6C57HW5BN2O7MW: 941.28Compound 21

[0367] (4-(3-((4-((8-((3-hexylnonyl)oxy)-8-oxooctyl)(8-oxo-8-(tridecan-5-yloxy)octyl) amino)butyl)amino)-3-oxopropyl)phenyl)boronic acid (Compound 21). UPLC-MS: m / z (ESI) for C57H105BN2O7: [M+H]+expected: 942.28; observed: 943.36. *HNMR (400 MHz, CDCh) 5 7.96 - 7.81 (m, 1H), 7.66 (d, J= 7.5 Hz, 1H), 7.15 (d, J= 10.2 Hz, 2H), 4.85 (s, 1H), 4.08 (d, J = 7.6 Hz, 2H), 3.51 (s, 4H), 3.17 (d, J = 6.5 Hz, 2H), 2.95 (d, J= 7.1 Hz, 2H), 2.61 -2.31 (m, 8H), 2.26 (q, J = 7.5 Hz, 4H), 1.72 - 1.36 (m, 12H), 1.26 (d, J = 12.0 Hz, 54H), 0.87 (t, J= 6.5 Hz, 12H).C58H104N2O5MW: 925.48Compound 24

[0368] 3-hexylnonyl 8-((4-(4-oxo-4-phenylbutanamido)butyl)(8-oxo-8-(tridecan-5-yloxy)octyl)amino)octanoate (Compound 24). UPLC-MS: m / z (ESI) for C58H104N2O6: [M+H]+expected: 926.48; observed: 928.16. 'H NMR (400 MHz, CDCh) 57.98 (d, J= 7.7 Hz, 1H), 7.69 - 7.28 (m, 3H), 7.26 (s, 1H), 6.37 (s, 1H), 4.86 (t, J= 6.4 Hz, 1H), 4.07 (t, J= 7.1 Hz, 2H), 3.36 (t, J= 6.7 Hz, 1H), 3.25 (q, J= 6.4 Hz, 1H), 2.60 (t, J= 6.7 Hz, 1H), 2.52 - 2.33 (m, 6H), 2.27 (t, J= 7.6 Hz, 4H), 1.78 - 1.38 (m, 24H), 1.28 (d, J= 19.1 Hz, 47H), 0.96 - 0.77 (m, 12H).CgrH^jBrNjOgMW: 992.36Compound 25

[0369] 3-hexylnonyl 8-((4-(3-(2 -bromophenoxy )propanamido)butyl)(8-oxo-8-(tridecan-5-yloxy)octyl)amino)octanoate (Compound 25). UPLC-MS: m / z (ESI) for CsrHiosBr^Oe:- 127 - LEGAL\113833648\6[M+H]+expected: 993.36; observed: 992.18. 'H NMR (400 MHz, CDCh) 87.53 (d, J= 7.9 Hz, 1H), 7.35 - 7.21 (m, 1H), 6.93 (d, J = 8.2 Hz, 1H), 6.86 (t, J = 7.7 Hz, 1H), 4.86 (t, J = 6.3 Hz, 1H), 4.30 (t, J= 5.9 Hz, 2H), 4.07 (t, J= 7.1 Hz, 2H), 3.30 (d, J= 6.3 Hz, 2H), 2.72 (t, J= 5.9 Hz, 2H), 2.39 (s, 6H), 2.27 (t, J= 7.6 Hz, 4H), 1.72 - 1.37 (m, 17H), 1.28 (d, J= 20.9 Hz, 51H), 0.87 (d, J = 7.2 Hz, 12H).CS7H103FN2O6MW: 931.46Compound 26

[0370] 3-hexylnonyl 8-((4-(3-(4-fluorophenoxy)propanamido)butyl)(8-oxo-8-(tridecan-5-yloxy)octyl)amino)octanoate (Compound 26). UPLC-MS: m / z (ESI) for C57H103FN2O6: [M+H]+expected: 932.46; observed: 933.51. 'H NMR (400 MHz, CDCh) 86.96 (t, J= 8.5 Hz, 2H), 6.84 (dd, J= 9.2, 4.3 Hz, 2H), 4.85 (d, J= 6.3 Hz, 1H), 4.22 (t, J= 6.2 Hz, 2H), 4.07 (t, J= 7.1 Hz, 2H), 3.29 (q, J= 6.2 Hz, 2H), 2.63 (t, J= 6.2 Hz, 2H), 2.49 (s, 6H), 2.27 (t, J= 7.6 Hz, 4H), 1.55 (ddt, J=28.8, 13.3, 6.8 Hz, 20H), 1.27 (d, J= 18.1 Hz, 47H), 0.98 - 0.75 (m, 12H).CgyHiojINjOgMW: 1039.36Compound 27

[0371] 3-hexylnonyl 8-((4-(3-(4-iodophenoxy)propanamido)butyl)(8-oxo-8-(tridecan-5-yloxy)octyl)amino)octanoate (Compound 27). UPLC-MS: m / z (ESI) for C57H103IN2O6: [M+H]+expected: 1040.36; observed: 1041.55. 'H NMR (400 MHz, CDCh) 87.63 - 7.49 (m, 2H), 6.77 -6.58 (m, 2H), 4.86 (t, J= 6.2 Hz, 1H), 4.23 (t, J= 6.2 Hz, 2H), 4.07 (t, J= 7.1 Hz, 2H), 3.28 (d, J = 6.0 Hz, 2H), 2.63 (t, J= 6.2 Hz, 2H), 2.47 (s, 6H), 2.28 (t, J= 7.5 Hz, 4H), 1.55 (tt, J= 21.8, 6.7 Hz, 18H), 1.36 - 1.12 (m, 57H), 0.88 (td, J= 7.0, 2.6 Hz, 12H).C37H1Q5N3O5MW: 912.48Compound 28

[0372] 3-hexylnonyl 8-((8-oxo-8-(tridecan-5-yloxy)octyl)(4-(3-(phenylamino)propanamido) butyl)amino)octanoate (Compound 28). UPLC-MS: m / z (ESI) for C57H105N3O5: [M+H]+- 128 - LEGAL\113833648\6expected: 913.48; observed: 913.20. 'H NMR (400 MHz, CDCh) 87.18 - 7.07 (m, 2H), 6.70 (t, J= 7.3 Hz, 1H), 6.67 - 6.58 (m, 2H), 4.86 (p, J= 6.3 Hz, 1H), 4.07 (t, J= 7.1 Hz, 2H), 3.45 (t, J = 6.1 Hz, 2H), 3.25 (q, J= 6.3 Hz, 2H), 2.59 - 2.34 (m, 6H), 2.35 - 2.17 (m, 4H), 1.69 - 1.37 (m, 17H), 1.28 (d, J= 20.4 Hz, 52H), 0.88 (td, J= 6.9, 2.7 Hz, 12H).C571bo4N20SSMW: 945.53Compound 29

[0373] 3-hexylnonyl 8-((8-oxo-8-(tridecan-5-yloxy)octyl)(4-(((2-(phenylthio)ethoxy) carbonyl)amino)butyl)amino)octanoate (Compound 29). Prepared using 2-(phenylthio)ethyl chloroformate (CAS 65935-47-9). UPLC-MS: m / z (ESI) for C57H104N2O6S: [M+H]+expected: 946.53; observed: 947.56. 'H NMR (400 MHz, CDCh) 88.17 - 8.02 (m, 1H), 7.40 - 7.32 (m, 1H), 7.30 - 7.22 (m, 1H), 7.20 - 7.12 (m, 1H), 6.75 (dt, J= 9.1, 2.8 Hz, 2H), 4.84 (p, J= 6.3 Hz, 1H), 4.19 (t, J = 6.9 Hz, 2H), 4.05 (t, J= 7.1 Hz, 2H), 3.12 (dt, J= 13.7, 6.5 Hz, 4H), 2.56 -2.42 (m, 6H), 2.25 (t, J= 7.5 Hz, 4H), 1.70 - 1.32 (m, 16H), 1.25 (d, J= 20.0 Hz, 51H), 0.85 (td, J = 6.9, 2.7 Hz, 12H).C57H toaBfNjOgMW: 992.36Compound 30

[0374] 3-hexylnonyl 8-((4-(3-(3-bromophenoxy)propanamido)butyl)(8-oxo-8-(tridecan-5- yloxy)octyl)amino)octanoate (Compound 30). UPLC-MS: m / z (ESI) for CsrHiosBrNfeOe:[M+H]+expected: 993.36; observed: 995.18. 'H NMR (400 MHz, CDCh) 87.16 - 7.02 (m, 3H), 6.89 - 6.77 (m, 1H), 4.92 -4.71 (m, 1H), 4.26 (t, J= 6.1 Hz, 2H), 4.07 (t, J= 7.1 Hz, 2H), 3.33 (d, J= 6.6 Hz, 2H), 2.68 (m, 8H), 2.28 (td, J= 7.5, 1.2 Hz, 4H), 1.55 (ddt, J= 28.9, 13.1, 6.8 Hz, 24H), 1.29 (d, J= 27.9 Hz, 51H), 0.88 (td, J= 6.1, 3.4 Hz, 12H).C56H104N2O5SMW; 917.52Compound 31

[0375] 3-hexylnonyl 8-((8-oxo-8-(tridecan-5-yloxy)octyl)(4-(4-(thiophen-3-yl)butanamido)- 129 - LEGAL\113833648\6butyl)amino)octanoate (Compound 31). UPLC-MS: m / z (ESI) for C56H104N2O5S: [M+H]+expected: 918.52; observed: 919.50. 'H NMR (400 MHz, CDCL) 87.32 - 7.19 (m, 1H), 7.00 - 6.87 (m, 2H), 6.11 (s, 1H), 4.96 - 4.77 (m, 1H), 4.07 (t, J= 7.1 Hz, 2H), 3.24 (q, J= 6.3 Hz, 2H), 2.67 (t, J= 7.5 Hz, 2H), 2.39 (s, 6H), 2.33 -2.21 (m, 4H), 2.16 (t, J= 7.5 Hz, 2H), 2.05 - 1.93 (m, 2H), 1.74 - 1.36 (m, 19H), 1.35 - 1.11 (m, 48H), 0.88 (td, J= 6.9, 2.6 Hz, 12H).C58H103H3OSMW: 938.48Compound 32

[0376] 3-hexylnonyl 8-((4-(3-(4-cyanophenoxy)propanamido)butyl)(8-oxo-8-(tridecan-5- yloxy)octyl)amino)octanoate (Compound 32). UPLC-MS: m / z (ESI) for C C58H103N3O6:[M+H]+expected: 939.48; observed: 940.11.XH NMR (400 MHz, CDCI3) 87.67 - 7.50 (m, 2H), 7.01 - 6.85 (m, 2H), 6.64 (s, 1H), 4.86 (s, OH), 4.32 (t, J= 6.2 Hz, 2H), 4.07 (t, J= 7.1 Hz, 2H), 3.28 (d, J= 5.9 Hz, 2H), 2.64 (t, J= 6.2 Hz, 2H), 2.40 (s, 6H), 2.27 (t, J= 7.5 Hz, 4H), 1.73 - 1.35 (m, 17H), 1.37 - 1.06 (m, 50H), 0.88 (td, J= 6.9, 2.6 Hz, 12H).MW: 917.52Compound 33

[0377] 3-hexylnonyl 8-((8-oxo-8-(tridecan-5-yloxy)octyl)(4-(4-(thiophen-2-yl)butanamido) butyl)amino)octanoate (Compound 33). UPLC-MS: m / z (ESI) for C56H104N2O5S: [M+H]+expected: 918.52; observed: 920.10. 'H NMR (400 MHz, CDC13) 87.11 (dd, J= 5.1, 1.2 Hz, 1H), 6.91 (dd, J = 5.1, 3.4 Hz, 1H), 6.79 (d, J= 3.7Hz, 1H), 6.13 (s, 1H), 4.86 (p, J = 6.2 Hz, 1H), 4.07 (t, J= 7.1 Hz, 2H), 3.24 (q, J = 6.3 Hz, 2H), 2.87 (t, J = 7.3 Hz, 2H), 2.38 (dt, J= 14.9, 7.3 Hz, 6H), 2.32 - 2.23 (m, 4H), 2.19 (t, J= 7.4 Hz, 2H), 2.03 (q, J= 7.5 Hz, 2H), 1.76 - 1.37 (m, 19H), 1.36 - 1.14 (m, 48H), 0.88 (td, J= 6.8, 2.6 Hz, 12H).Cgg H 10oBrNjOgSMW: 996.41Compound 35

[0378] 3-hexylnonyl 8-((4-(4-(5-bromothiophen-2-yl)butanamido)butyl)(8-oxo-8-(tridecan-5-- 130 - LEGAL\113833648\6yloxy)octyl)amino)octanoate (Compound 35). UPLC-MS: m / z (ESI) for CseHiosBr^OsS: [M+H]+expected: 997.41; observed: 999.34. 'H NMR (400 MHz, CDC13) 88.19 (d, J= 1.7 Hz, 1H), 7.00 (d, J = 6.1 Hz, 1H), 4.85 (p, J = 6.3 Hz, 1H), 4.07 (t, J = 7.1 Hz, 2H), 3.33 (dq, J = 22.3, 6.4 Hz, 2H), 2.88 (s, 6H), 2.28 (td, J= 7.4, 1.5 Hz, 4H), 1.86 (s, 4H), 1.79 - 1.42 (m, 18H), 1.30 (d, J= 38.9 Hz, 54H), 0.88 (td, J= 5.9, 3.4 Hz, 12H).o s'" '<... -■ J 'O'... '-xA,N-NzO S' OK a.. Ji.. N....... ji i....,,..HC53H102N5O5SMW: 935.50Compound 38

[0379] 3-hexylnonyl 8-((4-(3-((l-methyl-l / f-tetrazol-5-yl)thio)propanamido)butyl)(8-oxo-8-(tridecan-5-yloxy)octyl)amino)octanoate (Compound 38). UPLC-MS: m / z (ESI) for C53H102N6O5S: [M+H]+expected: 936.50; observed: 938.11. 'H NMR (400 MHz, CDC13) 86.57 (s, 1H), 4.86 (p, J= 6.3 Hz, 1H), 4.07 (t, J= 7.1 Hz, 2H), 3.91 (s, 3H), 3.58 (t, J= 6.6 Hz, 2H), 3.26 (q, J= 6.2 Hz, 2H), 2.78 (d, J= 6.6 Hz, 2H), 2.38 (q, J= 8.1 Hz, 6H), 2.27 (t, J= 7.5 Hz, 4H), 1.77- 1.36 (m, 23H), 1.27 (d, J = 18.4 Hz, 54H), 0.99 - 0.70 (m, 12H).C53H102N6O5MW: 903.44Compound 39

[0380] 3-hexylnonyl 8-((4-(4-(l / f-tetrazol-l-yl)butanamido)butyl)(8-oxo-8-(tridecan-5-yloxy)octyl)amino)octanoate (Compound 39). PLC-MS: m / z (ESI) for C53H102N6O5: [M+H]+expected: 904.44; observed: 905.94. 'H NMR (400 MHz, CDCh) 88.68 (s, 1H), 6.51 (s, 1H), 4.98 - 4.76 (m, 1H), 4.58 (t, J= 6.7 Hz, 2H), 4.07 (t, J= 7.1 Hz, 2H), 3.24 (q, J= 6.3 Hz, 2H), 2.38 (dt, J= 11.5, 7.2 Hz, 6H), 2.32 -2.22 (m, 6H), 2.16 (t, 7 = 6.7 Hz, 2H), 1.73 - 1.45 (m, 18H), 1.36 - 1.10 (m, 50H), 0.88 (td, 7= 6.9, 2.6 Hz, 12H).C60H110N2O8MW: 987.55Compound 40

[0381] 3-hexylnonyl 8-((8-oxo-8-(tridecan-5-yloxy)octyl)(4-(3-(3,4,5-trimethoxyphenyl) propanamido)butyl)amino)octanoate (Compound 40). UPLC-MS: m / z (ESI) for C60H110N2O8:- 131 - LEGAL\113833648\6[M+H]+expected: 988.55; observed: 990.18. 'H NMR (400 MHz, CDC13) 86.42 (s, 2H), 6.00 (s, 1H), 4.86 (p, J= 6.2 Hz, 1H), 4.07 (t, J= 7.1 Hz, 2H), 3.82 (d, J = 8.3 Hz, 8H), 3.23 (q, J= 6.3 Hz, 2H), 2.90 (dd, J= 8.6, 6.8 Hz, 2H), 2.43 (dd, J= 8.6, 6.9 Hz, 2H), 2.41 - 2.32 (m, 7H), 2.27 (t, J= 7.3 Hz, 4H), 1.76 - 1.35 (m, 21H), 1.34 - 1.09 (m, 47H), 0.88 (td, J= 6.9, 2.5 Hz, 12H).0C57H104N2O7MW: 929.47Compound 41

[0382] 3-hexylnonyl 8-((8-oxo-8-(tridecan-5-yloxy)octyl)(4-(((2-phenoxyethoxy)carbonyl) amino)butyl)amino)octanoate (Compound 41). Prepared using 2-phenoxyethyl chloroformate (CAS 34743-87-8). UPLC-MS: m / z (ESI) for C57H104N2O7: [M+H]+expected: 930.47; observed: 931.81.XH NMR (400 MHz, CDCh) 87.39 - 7.18 (m, 2H), 7.04 - 6.86 (m, 3H), 5.46 (s, 1H), 4.86 (p, J= 6.3 Hz, 1H), 4.41 (t, J= 4.7 Hz, 2H), 4.15 (t, J = 4.7 Hz, 2H), 4.07 (t, J= 7.1 Hz, 2H), 3.18 (d, J= 6.2 Hz, 2H), 2.45 - 2.33 (m, 6H), 2.27 (t, J= 7.5 Hz, 4H), 1.71 - 1.36 (m, 29H), 1.27 (d, J= 18.5 Hz, 48H), 0.88 (td, J= 6.9, 2.6 Hz, 12H).MW: 925.52Compound 44

[0383] 3-hexylnonyl 8-((8-oxo-8-(tridecan-5-yloxy)octyl)(4-(4-(p-tolyl)butanamido)butyl) amino)octanoate (Compound 44). UPLC-MS: m / z (ESI) for C59H108N2O5: [M+H]+expected: 926.52; observed: 928.16.XH NMR (400 MHz, CDCI3) 87.15 - 7.02 (m, 4H), 6.05 (s, 1H), 4.86 (p, J= 6.3 Hz, 1H), 4.07 (t, J= 7.1 Hz, 2H), 3.23 (q, J= 6.2 Hz, 2H), 2.61 (t, J= 7.5 Hz, 2H), 2.37 (s, 6H), 2.33 -2.23 (m, 7H), 2.14 (t, J= 7.5 Hz, 2H), 1.95 (q, J= 7.5 Hz, 2H), 1.74 - 1.37 (m, 18H), 1.28 (d, J= 20.0 Hz, 50H), 0.88 (td, J= 6.8, 2.9 Hz, 12H).C58H105ClN2O5MW: 945.94Compound 45

[0384] 3-hexylnonyl 8-((4-(4-(4-chlorophenyl)butanamido)butyl)(8-oxo-8-(tridecan-5-- 132 - LEGAL\113833648\6yloxy)octyl)amino)octanoate (Compound 45). UPLC-MS: m / z (ESI) for C58H105CIN2O5:[M+H]+expected: 946.94; observed: 945.31. 'H NMR (400 MHz, CDCh) 87.24 (d, J= 13.6 Hz, 2H), 7.10 (d, J= 8.2 Hz, 2H), 6.13 (s, 1H), 4.86 (p, J= 6.3 Hz, 1H), 4.07 (t, J= 7.1 Hz, 2H), 3.23 (d, J= 6.0 Hz, 2H), 2.62 (t, J= 7.6 Hz, 2H), 2.37 (dt, J= 15.1, 7.2 Hz, 6H), 2.28 (t, J= 7.5 Hz, 4H), 2.13 (t, J = 7.4 Hz, 2H), 1.95 (q, J = 7.5 Hz, 2H), 1.81 - 1.36 (m, 19H), 1.28 (d, J = 19.8 Hz, 48H), 0.88 (td, J= 6.8, 2.9 Hz, 12H).C57H ’ 030^20^3MW: 995.97Compound 46

[0385] 3 -hexylnonyl 8-((4-(3-((4-chlorophenyl)sulfonyl)propanamido)butyl)(8-oxo-8- (tridecan-5-yloxy)octyl)amino)octanoate (Compound 46). UPLC-MS: m / z (ESI) for C57H103CIN2O7S: [M+H]+expected: 996.97; observed: 995.18. 'H NMR (400 MHz, CDCh) 8 7.85 (d, J= 8.5 Hz, 2H), 7.65 - 7.45 (m, 2H), 6.78 (s, 1H), 4.86 (q, J= 6.3 Hz, 1H), 4.08 (t, J = 7.1 Hz, 2H), 3.46 (t, J= 7.7Hz, 2H), 3.18 (q, J= 6.0 Hz, 2H), 2.61 (t, J= 7.7Hz, 2H), 2.39 (s, 6H), 2.28 (t, J= 7.5 Hz, 4H), 1.80 - 1.37 (m, 18H), 1.27 (d, J= 15.3 Hz, 49H), 0.88 (td, J= 6.7, 2.8 Hz, 12H).C58H106N2O7SMW: 975.55Compound 47

[0386] 3-hexylnonyl 8-((8-oxo-8-(tridecan-5-yloxy)octyl)(4-(3-tosylpropanamido)butyl) amino)octanoate (Compound 47). UPLC-MS: m / z (ESI) for C58H106N2O7S: [M+H]+expected: 976.55; observed: 978.68. 'H NMR (400 MHz, CDCh) 87.78 (d, J= 7.9 Hz, 2H), 7.35 (d, J= 7.9 Hz, 2H), 6.68 (s, 1H), 4.87 (t, J = 6.2 Hz, 1H), 4.08 (t, J= 7.1 Hz, 2H), 3.57-3.40 (m, 2H), 3.18 (q, J= 6.1 Hz, 2H), 2.59 (t, J= 7.8 Hz, 2H), 2.45 (s, 3H), 2.38 (t, J= 7.5 Hz, 6H), 2.28 (t, J= 7.5 Hz, 4H), 1.77- 1.37 (m, 18H), 1.28 (d, J= 24.4 Hz, 49H), 0.87 (q, J= 4.5 Hz, 12H).- 133 - LEGAL\113833648\6C57H103BrN2O7SMW: 1040.42Compound 48

[0387] 3 -hexylnonyl 8-((4-(3-((4-bromophenyl)sulfonyl)propanamido)butyl)(8-oxo-8-(tridecan-5-yloxy)octyl)amino)octanoate (Compound 48). UPLC-MS: m / z (ESI) for C57Hio3BrN207S: [M+H]+expected: 1041.42; observed: 1043.70. 'HNMR (400 MHz, CDC13) 8 7.85 - 7.62 (m, 4H), 6.77 (s, 1H), 4.87 (t, J= 6.2 Hz, 1H), 4.08 (t, J= 7.1 Hz, 2H), 3.46 (t, J= 7.7 Hz, 2H), 3.17 (d, J= 5.9 Hz, 2H), 2.60 (t, J= 7.7 Hz, 2H), 2.38 (s, 6H), 2.28 (t, J= 7.5 Hz, 4H), 1.71 - 1.37 (m, 17H), 1.27 (d, J = 15.2 Hz, 50H), 0.97- 0.72 (m, 12H).C57H103BrN2O5SMW; 1008.42Compound 49

[0388] 3-hexylnonyl 8-((4-(3-((4-bromophenyl)thio)propanamido)butyl)(8-oxo-8-(tridecan-5-yloxy)octyl)amino)octanoate (Compound 49). UPLC-MS: m / z (ESI) for CsrHiosBrlSbOsS: [M+H]+expected: 1009.42; observed: 1011.94. 'HNMR (400 MHz, CDC13) 87.50 - 7.33 (m, 2H), 7.25 - 7.12 (m, 2H), 6.46 (s, 1H), 4.98 - 4.68 (m, 1H), 4.07 (t, J= 7.1 Hz, 2H), 3.22 (p, J = 6.7 Hz, 4H), 2.61 - 2.33 (m, 8H), 2.28 (t, J= 7.5 Hz, 4H), 1.80 - 1.35 (m, 15H), 1.28 (d, J= 20.2 Hz, 52H), 0.88 (dq, J= 7.0, 2.9 Hz, 12H).C57H103CIN2O5SMW: 963.97Compound 50

[0389] 3-hexylnonyl 8-((4-(3-((4-chlorophenyl)thio)propanamido)butyl)(8-oxo-8-(tridecan-5-yloxy)octyl)amino)octanoate (Compound 50). UPLC-MS: m / z (ESI) for C57H103CIN2O5S:[M+H]+expected: 964.97; observed: 965.42. 'H NMR (400 MHz, CDCh) 87.26 (d, J= 3.4 Hz, 4H), 6.45 (s, 1H), 4.86 (p, J= 6.3 Hz, 1H), 4.07 (t, J= 7.1 Hz, 2H), 3.22 (dt, J= 15.0, 6.8 Hz, 4H), 2.40 (dt, J = 28.1, 7.4 Hz, 8H), 2.28 (t, J= 7.5 Hz, 4H), 1.79 - 1.35 (m, 17H), 1.28 (d, J = 20.1 Hz, 50H), 0.88 (td, J= 6.9, 2.9 Hz, 12H).- 134 - LEGAL\113833648\6C58H106N2O5SMW: 943.55Compound 51

[0390] 3-hexylnonyl 8-((8-oxo-8-(tridecan-5-yloxy)octyl)(4-(3-?-tolylthio)propanamido) butyl)amino)octanoate (Compound 51). UPLC-MS: m / z (ESI) for C58H106N2O5S: [M+H]+expected: 944.55; observed: 946.56. 'H NMR (400 MHz, CDCh) 57.35 - 7.23 (m, 2H), 7.10 (d, J = 7.8 Hz, 2H), 6.33 (d, J = 6.9 Hz, 1H), 4.86 (p, J= 6.3 Hz, 1H), 4.07 (t, J = 7.1 Hz, 2H), 3.19 (dt, J= 23.6, 6.7 Hz, 4H), 2.39 (dt, J= 19.9, 5.4 Hz, 9H), 2.32 (s, 3H), 2.27 (t, J= 7.5 Hz, 4H), 1.77 - 1.34 (m, 22H), 1.33 (m, 55H), 0.88 (td, J= 6.7, 2.9 Hz, 12H).C56H103N3O5MW; 900.47Compound 55

[0391] 3-hexylnonyl 8-((4-(4-(U7-pyrrol-l-yl)butanamido)butyl)(8-oxo-8-(tridecan-5- yloxy)octyl)amino)octanoate (Compound 55). UPLC-MS: m / z (ESI) for C56H105N3O5: [M+H]+expected: 901.47; observed: 903.09. 'H NMR (400 MHz, CDCh) 86.63 (t, J= 2.1 Hz, 2H), 6.13 (t, J= 2.1 Hz, 2H), 4.86 (p, J= 6.3 Hz, 1H), 4.08 (t, J= 7.1 Hz, 2H), 4.02 - 3.88 (m, 2H), 3.23 (q, J= 6.2 Hz, 2H), 2.38 (s, 6H), 2.33 - 2.20 (m, 4H), 2.09 (dd, J= 9.2, 5.2 Hz, 4H), 1.76 - 1.37 (m, 19H), 1.37- 1.13 (m, 50H), 0.88 (td, J= 6.7, 2.8 Hz, 12H).C55H104N4O5MW: 901.46Compound 56

[0392] 3-hexylnonyl 8-((4-(4-(l / f-imidazol-l-yl)butanamido)butyl)(8-oxo-8-(tridecan-5- yloxy)octyl)amino)octanoate (Compound 56). UPLC-MS: m / z (ESI) for C55H104N4O5: [M+H]+expected: 902.46; observed: 903.14. 'H NMR (400 MHz, CDCh) 87.46 (s, 1H), 7.06 (s, 1H), 6.91 (s, 1H), 6.25 (s, 1H), 4.94 -4.75 (m, 1H), 4.19 - 3.93 (m, 4H), 3.24 (q, J = 6.2 Hz, 2H), 2.38 (dt, J= 14.9, 7.3 Hz, 6H), 2.28 (t, J= 7.5 Hz, 4H), 2.18 -2.03 (m, 4H), 1.83 - 1.37 (m, 18H), 1.27 (d, J = 15.0 Hz, 49H), 0.88 (td, J = 6.7, 2.9 Hz, 12H).- 135 - LEGAL\113833648\6HC55H104N4O5MW: 901,46Compound 57

[0393] 3-hexylnonyl 8-((4-(4-(l / f-pyrazol-l-yl)butanamido)butyl)(8-oxo-8-(tridecan-5-yloxy)octyl)amino)octanoate (Compound 57). UPLC-MS: m / z (ESI) for C55H104N4O5: [M+H]+expected: 902.46; observed: 904.09. 'H NMR (400 MHz, CDCh) 87.49 (d, J= 1.9 Hz, 1H), 7.39 (d, J = 2.3 Hz, 1H), 6.52 (s, 1H), 6.24 (t, J = 2.1 Hz, 1H), 4.86 (p, J = 6.3 Hz, 1H), 4.21 (t, J = 6.2 Hz, 2H), 4.07 (t, J= 7.1 Hz, 2H), 3.25 (q, J= 6.2 Hz, 2H), 2.37 (s, 5H), 2.28 (t, J= 7.5 Hz, 4H), 2.14 (dt, J= 16.9, 6.2 Hz, 4H), 1.78 - 1.36 (m, 20H), 1.27 (d, J= 17.4 Hz, 47H), 0.88 (td, J= 6.8, 2.8 Hz, 12H).C54H103N5O5MW: 902.45Compound 58

[0394] 3-hexylnonyl 8-((4-(4-(U7-l,2,4-triazol-l-yl)butanamido)butyl)(8-oxo-8-(tridecan-5-yloxy)octyl)amino)octanoate (Compound 58). UPLC-MS: m / z (ESI) for C54H103N5O5: [M+H]+expected: 903.45; observed: 904.79.XH NMR (400 MHz, CDCh) 88.09 (s, 1H), 7.94 (s, 1H), 6.39 (s, 1H), 4.95 - 4.70 (m, 1H), 4.29 (t, J= 6.4 Hz, 2H), 4.07 (t, J= 7.1 Hz, 2H), 3.24 (q, J = 6.3 Hz, 2H), 2.38 (dt, J= 14.7, 7.3 Hz, 6H), 2.28 (t, J= 7.5 Hz, 4H), 2.23 -2.07 (m, 4H), 1.75 -1.36 (m, 17H), 1.27 (d, J= 15.4 Hz, 50H), 0.88 (td, J= 6.8, 2.8 Hz, 12H).OHC65H112N2O7MW: 1033.62Compound 59

[0395] 3-hexylnonyl 8-((4-(4,4-bis(4-hydroxyphenyl)pentanamido)butyl)(8-oxo-8-(tridecan-5-yloxy)octyl)amino)octanoate (Compound 59). UPLC-MS: m / z (ESI) for C65H112N2O7: [M+H]+expected: 1034.62; observed: 1033.85.XH NMR (400 MHz, CDCh) 86.97 (d, J = 8.2 Hz, 4H), 6.72 (d, J = 8.2 Hz, 4H), 6.20 (d, J = 5.6 Hz, 1H), 4.87 (s, 1H), 4.08 (t, J = 7.1 Hz, 2H), 3.12 (q, J- 136 - LEGAL\113833648\6= 5.9 Hz, 2H), 2.50 (s, 6H), 2.45 - 2.31 (m, 2H), 2.28 (t, J= 7.5 Hz, 5H), 2.03 - 1.85 (m, 2H), 1.72 - 1.36 (m, 19H), 1.26 (d, J= 9.8 Hz, 53H), 0.87 (dq, J= 7.0, 2.9 Hz, 12H).MW: 943.49Compound 640

[0396] 3 -hexylnonyl 8-((4-(3-(4-(hydroxymethyl)phenoxy)propanamido)butyl)(8-oxo-8-(tridecan-5-yloxy)octyl)amino)octanoate (Compound 640). UPLC-MS: m / z (ESI) for C58H106N2O7: [M+H]+expected: 944.49; observed: 944.96. ' H NMR (400 MHz, CDCh) 57.39 -7.22 (m, 2H), 7.03 - 6.81 (m, 3H), 4.86 (p, J= 6.2 Hz, 1H), 4.61 (s, 2H), 4.28 (t, J= 6.1 Hz, 2H), 4.07 (t, J= 7.1 Hz, 2H), 3.35 (d, J= 6.3 Hz, 2H), 2.95 (s, 6H), 2.70 (t, J= 6.1 Hz, 2H), 2.33 -2.17 (m, 4H), 1.74 (m, 4H), 1.56 (ddt, J= 28.2, 12.1, 6.7 Hz, 12H), 1.46 - 1.10 (m, 52H), 0.88 (td, J= 5.8, 3.4 Hz, 12H).C58H106N2O8MW: 971.50Compound 641

[0397] 3-hexylnonyl 8-((4-(3-((2,3-dihydrobenzo[ / >][l,4]dioxin-6-yl)oxy)propanamido)butyl) (8-oxo-8-(tridecan-5-yloxy)octyl)amino)octanoate (Compound 641). UPLC-MS: m / z (ESI) for C59H106N2O8: [M+H]+expected: 972.50; observed: 974.28.XH NMR (400 MHz, CDCh) 86.76 (d, J= 8.8 Hz, 1H), 6.53 - 6.32 (m, 3H), 4.86 (s,lH), 4.34 - 4.13 (m, 6H), 4.07 (t, J= 7.1 Hz, 2H), 3.27 (q, J= 6.2 Hz, 2H), 2.59 (t, J= 6.1 Hz, 2H), 2.38 (dt, J= 15.0, 7.3 Hz, 6H), 2.27 (t, J = 7.5 Hz, 4H), 1.77- 1.36 (m, 18H), 1.27 (d, J= 18.1 Hz, 49H), 0.88 (td, J= 6.9, 2.7 Hz, 12H).0" oC57H103N3O9SMW: 1006.52Compound 642

[0398] 3-hexylnonyl 8-((4-(3-((4-nitrophenyl)sulfonyl)propanamido)butyl)(8-oxo-8-(tridecan-5-yloxy)octyl)amino)octanoate (Compound 642). UPLC-MS: m / z (ESI) for C57H103N3O9S:[M+H]+expected: 1007.52; observed: 1008.84.XH NMR (400 MHz, CDCh) 88.41 (d, J= 8.7 Hz, 2H), 8.12 (d, J= 8.6 Hz, 2H), 6.89 (s, 1H), 4.87 (t, J= 6.2 Hz, 1H), 4.08 (t, J= 7.1 Hz, 2H),- 137 - LEGAL\113833648\63.53 (t, J = 7.5 Hz, 2H), 3.16 (q, J = 6.1 Hz, 2H), 2.67 (d, J = 7.6 Hz, 2H), 2.38 (t, J = 7.3 Hz, 6H), 2.29 (t, J= 7.5 Hz, 4H), 1.77- 1.38 (m, 15H), 1.36 - 1.10 (m, 52H), 0.88 (td, J= 5.9, 3.4 Hz, 12H).C59H104FN3O5MW: 970.49Compound 643

[0399] 3 -hexylnonyl 8-((4-(l-((4-fluorophenyl)carbamoyl)cyclopropane-l-carboxamido) butyl)(8-oxo-8-(tridecan-5-yloxy)octyl)amino)octanoate (Compound 643). UPLC-MS: m / z (ESI) for C59H104FN3O6: [M+H]+expected: 971.49; observed: 973.12. 'H NMR (400 MHz, CDCh) 8 7.53 (dd, J= 8.9, 4.8 Hz, 2H), 6.99 (t, J = 8.7 Hz, 2H), 6.51 (s, 1H), 4.86 (p, J = 6.3 Hz, 1H), 4.08 (t, J= 7.1 Hz, 2H), 3.27 (q, J= 6.1 Hz, 2H), 2.39 (d, J= 9.4 Hz, 6H), 2.28 (t, J= 7.5 Hz, 4H), 1.70 (q, J= 4.6 Hz, 2H), 1.67 - 1.36 (m, 16H), 1.27 (d, J= 17.5 Hz, 52H), 0.88 (td, J= 6.8, 2.8 Hz, 12H).MW: 986.58Compound 64

[0400] 3 -hexylnonyl 8-((4-(2-acetamido-3-(phenylthio)propanamido)butyl)(8-oxo-8-(tridecan-5-yloxy)octyl)amino)octanoate (Compound 644). UPLC-MS: m / z (ESI) for C59H107N3O6S:[M+H]+expected: 987.58; observed: 988.58. 'H NMR (400 MHz, CDCh) 87.50 - 7.40 (m, 2H), 7.31 (t, J = 7.6 Hz, 2H), 7.24 - 7.07 (m, 1H), 4.86 (s, 1H), 4.07 (t, J = 7.1 Hz, 2H), 3.38 (dd, J = 13.8, 5.4 Hz, 1H), 3.20 (s, 3H), 2.37 (m, 6H), 2.27 (t, J= 7.5 Hz, 4H), 1.97 (s, 3H), 1.55 (ddd, J = 28.9, 11.9, 6.7 Hz, 17H), 1.40 (s, 3H), 1.27 (d, J= 14.1 Hz, 50H), 0.88 (td, J= 6.9, 2.6 Hz, 12H).C63H108N2O8MW: 989.57Compound 645

[0401] 3-hexylnonyl 8-((4-(3-([l,l'-biphenyl]-4-yloxy)propanamido)butyl)(8-oxo-8-(tridecan- 5-yloxy)octyl)amino)octanoate (Compound 645). UPLC-MS: m / z (ESI) for C64H110N2O5: [M+H]+- 138 - LEGAL\113833648\6expected: 989.59; observed: 989.63. 'H NMR (400 MHz, CDC13) 87.63 - 7.55 (m, 2H), 7.54 - 7.47 (m, 2H), 7.44 - 7.38 (m, 2H), 7.37 - 7.29 (m, 1H), 7.25 (d, J= 8.3 Hz, 2H), 6.12 (s, 1H), 4.97 - 4.76 (m, 1H), 4.07 (t, J= 7.1 Hz, 2H), 3.25 (q, J= 6.2 Hz, 2H), 2.69 (t, J= 7.5 Hz, 2H), 2.38 (s, 5H), 2.27 (t, J = 7.5 Hz, 4H), 2.19 (t, J = 7.5 Hz, 2H), 2.09 - 1.98 (m, 2H), 1.70 - 1.36 (m, 16H), 1.27 (d, J= 19.3 Hz, 51H), 0.88 (td, J= 6.9, 2.7 Hz, 12H).HC57H104N2O5SMW: 929.53Compound 646

[0402] 3-hexylnonyl 8-((4-(2-(benzylthio)acetamido)butyl)(8-oxo-8-(tridecan-5-yloxy)octyl)amino)octanoate (Compound 646). UPLC-MS: m / z (ESI) for C57H104N2O5S: [M+H]+expected: 930.53; observed: 930.17. 'H NMR (400 MHz, CDCh) 87.44 - 7.09 (m, 5H), 4.86 (p, J= 6.3 Hz, 1H), 4.08 (t, J= 7.1 Hz, 2H), 3.74 (s, 2H), 3.20 (s, 2H), 3.13 (s, 2H), 2.95 (s, 2H), 2.37 (s, 6H), 2.28 (td, J =7.5, 1.3 Hz, 4H), 1.56 (tt, J= 21.9, 6.9 Hz, 17H), 1.28 (d, J= 24.3 Hz, 50H), 0.88 (td, J= 6.9, 2.5 Hz, 12H).C57H102FN3O6MW: 944.46Compound 647

[0403] 3-hexylnonyl 8-((4-(3-((4-fluorophenyl)amino)-3-oxopropanamido)butyl)(8-oxo-8- (tridecan-5-yloxy)octyl)amino)octanoate (Compound 647). UPLC-MS: m / z (ESI) for C57H102FN3O6: [M+H]+expected: 945.46; observed: 945.20. 'H NMR (400 MHz, CDCI3) 89.86 (s, 1H), 7.83 (s, 1H), 7.59 - 7.46 (m, 2H), 7.08 - 6.89 (m, 2H), 4.87 (p, J= 6.3 Hz, 1H), 4.08 (t, J = 7.1 Hz, 2H), 3.28 (d, J = 4.0 Hz, 4H), 2.42 (q, J= 6.6 Hz, 6H), 2.28 (t, J= 7.5 Hz, 4H), 1.70 - 1.37 (m, 15H), 1.28 (d, J= 19.6 Hz, 52H), 0.88 (td, J= 6.8, 2.9 Hz, 12H).C61H110N2O7MW: 983.56Compound 648

[0404] 3-hexylnonyl 8-((4-(3-((2,2-dimethyl-2,3-dihydrobenzofuran-7-yl)oxy)propanamido)- 139 - LEGAL\113833648\6butyl)(8-oxo-8-(tridecan-5-yloxy)octyl)amino)octanoate (Compound 648). UPLC-MS: m / z (ESI) for C61H110N2O7: [M+H]+expected: 984.56; observed: 986.20. 'H NMR (400 MHz, CDCh) 5 6.90 - 6.72 (m, 3H), 6.68 (s, 1H), 4.97 - 4.76 (m, 1H), 4.29 (t, J= 6.0 Hz, 2H), 4.07 (t, J= 7.1 Hz, 2H), 3.27 (d, J= 6.3 Hz, 2H), 3.03 (s, 2H), 2.65 (t, J= 6.0 Hz, 2H), 2.37 (q, J= 9.2 Hz, 6H), 2.30 - 2.17 (m, 4H), 1.67- 1.51 (m, 7H), 1.49 (s, 8H), 1.39 (s, 6H), 1.28 (d, J= 19.5 Hz, 52H), 0.88 (td, J= 6.9, 2.6 Hz, 12H).C57H102Cl2N2O6SMW: 1014.41Compound 79

[0405] 3-hexylnonyl 8-((4-(3-((2,5-dichlorophenyl)sulfinyl)propanamido)butyl)(8-oxo-8-(tridecan-5-yloxy)octyl)amino)octanoate (Compound 79). UPLC-MS: m / z (ESI) for C57H102CI2N2O6S: [M+H]+expected: 1015.41; observed: 1015.97. 'H NMR (400 MHz, CDCh) 8 7.80 (s, 1H), 7.40 (dd, J = 8.5, 2.5 Hz, 1H), 7.33 (d, J= 8.3 Hz, 1H), 6.79 (s, 1H), 4.86 (t, J= 6.2 Hz, 1H), 4.08 (t, J= 7.1 Hz, 2H), 3.45 (s, OH), 3.28 - 3.05 (m, 2H), 2.55 - 2.35 (m, 7H), 2.28 (t, J = 7.5 Hz, 4H), 1.59 (dt, J= 14.1, 7.0 Hz, 5H), 1.51 (q, J= 7.4 Hz, 6H), 1.48 - 1.36 (m, 6H), 1.28 (d, J= 20.8 Hz, 49H), 0.88 (td, J= 6.9, 2.5 Hz, 12H).C57H102Cl2N2O7SMW: 1030.41Compound 80

[0406] 3-hexylnonyl 8-((4-(3-((2,5-dichlorophenyl)sulfonyl)propanamido)butyl)(8-oxo-8-(tridecan-5-yloxy)octyl)amino)octanoate (Compound 80). UPLC-MS: m / z (ESI) for C57H102CI2N2O7S: [M+H]+expected: 1031.41; observed: 1031.97. 'H NMR (400 MHz, CDCh) 8 7.85 (s, 1H), 7.42 (dd, J = 8.5, 2.5 Hz, 1H), 7.31 (d, J= 8.3 Hz, 1H), 6.79 (s, 1H), 4.86 (t, J= 6.2 Hz, 1H), 4.08 (t, J= 7.1 Hz, 2H), 3.45 (s, OH), 3.28 - 3.05 (m, 2H), 2.55 - 2.35 (m, 7H), 2.28 (t, J = 7.5 Hz, 4H), 1.59 (dt, J= 14.1, 7.0 Hz, 5H), 1.51 (q, J= 7.4 Hz, 6H), 1.48 - 1.36 (m, 6H), 1.28 (d, J= 20.8 Hz, 49H), 0.88 (td, J= 6.9, 2.5 Hz, 12H).- 140 - LEGAL\113833648\6C55H104N4O5SMW: 933.52Compound 81

[0407] 3-hexylnonyl 8-((4-(3-((l-methyl-l / f-imidazol-2-yl)thio)propanamido)butyl)(8-oxo-8- (tridecan-5-yloxy)octyl)amino)octanoate (Compound 81). UPLC-MS: m / z (ESI) for C55H104N4O5S: [M+H]+expected: 934.52; observed: 934.05. 'H NMR (400 MHz, CDCI3) 86.85 (d, J= 2.4 Hz, 2H), 6.65 (d, J= 2.3 Hz, 1H), 4.85 (s, OH), 4.34 (t, J= 6.4 Hz, 2H), 4.07 (t, J= 7.1 Hz, 2H), 3.68 - 3.52 (m, 3H), 3.26 (d, J= 6.5 Hz, 2H), 2.91 (s, 6H), 2.77 (t, J= 6.4 Hz, 2H), 2.28 (t, J= 7.5 Hz, 4H), 1.78 (d, J= 37.6 Hz, 3H), 1.56 (dt, J= 21.5, 12.4 Hz, 13H), 1.30 (d, J= 39.4 Hz, 50H), 0.88 (tq, J= 6.8, 2.4 Hz, 12H).C55H105N5O5MW: 916.47Compound 82

[0408] 3-hexylnonyl 8-((4-(4-(l-methyl-l / f-l,2,4-triazol-5-yl)butanamido)butyl)(8-oxo-8- (tridecan-5-yloxy)octyl)amino)octanoate (Compound 82). UPLC-MS: m / z (ESI) for C55H105N5O5: [M+H]+expected: 917.47; observed: 917.09.XH NMR (400 MHz, CDCh) 88.04 (s, 1H), 6.41 (s, 1H), 4.96 - 4.73 (m, 1H), 4.07 (t, J= 7.1 Hz, 2H), 3.64 (s, 3H), 3.23 (q, J= 6.3 Hz, 2H), 2.84 (t, J= 7.1 Hz, 2H), 2.37 (q, J= 8.5 Hz, 6H), 2.28 (q, J= 7.0 Hz, 6H), 2.10 (q, J= 6.9 Hz, 2H), 1.81 - 1.37 (m, 18H), 1.36 - 1.10 (m, 49H), 0.88 (td, J= 6.9, 2.6 Hz, 12H).C57H103FN2O7SMW: 979.52Comnound 83

[0409] 3-hexylnonyl 8-((4-(3-((2-fluorophenyl)sulfonyl)propanamido)butyl)(8-oxo-8- (tridecan-5-yloxy)octyl)amino)octanoate (Compound 83). UPLC-MS: m / z (ESI) for C57H103FN2O7S: [M+H]+expected: 980.52; observed: 980.01. 'H NMR (400 MHz, CDCh) 87.92 - 7.76 (m, 1H), 7.58 - 7.37 (m, 3H), 7.02 (d, J= 6.0 Hz, H), 4.85 (d, J= 6.2 Hz, 1H), 4.07 (dd, J = 7.9, 6.0 Hz, 4H), 3.30 (q, J= 6.3 Hz, 2H), 2.39 (q, J= 7.3 Hz, 6H), 2.28 (t, J= 7.5 Hz, 4H), 1.73 - 1.47 (m, 20H), 1.27 (d, J= 19.8 Hz, 50H), 0.88 (td, J= 6.9, 2.7 Hz, 12H).- 141 - LEGAL\113833648\6C59H106N2O9SMW: 1019.56Compound 84

[0410] 3-hexylnonyl 8-((4-(3-((2,3-dihydrobenzo[ / >][l,4]dioxin-5-yl)sulfonyl)propanamido) butyl)(8-oxo-8-(tridecan-5-yloxy)octyl)amino)octanoate (Compound 84). UPLC-MS: m / z (ESI) for C59H106N2O9S: [M+H]+expected: 1020.56; observed: 1020.12. 'H NMR (400 MHz, CDCh) 8 7.54 - 7.31 (m, 2H), 6.99 (d, J= 8.3 Hz, 1H), 4.86 (s, OH), 4.44 - 4.24 (m, 4H), 4.08 (t, J= 7.1 Hz, 2H), 3.49 (s, 4H), 3.47 - 3.35 (m, 2H), 3.21 (s, 2H), 2.60 (t, J= 7.7 Hz, 2H), 2.43 (s, 6H), 2.31 -2.23 (m, 4H), 1.56 (ddt, J= 28.8, 13.6, 6.1 Hz, 12H), 1.28 (d, J= 23.2 Hz, 52H), 0.88 (tt, J = 7.1, 1.8 Hz, 12H).C60H108N2O9SMW: 1033.59Compound 85

[0411] 3-hexylnonyl 8-((4-(3-((3,4-dihydro-2 / f-benzo[ / >][l,4]dioxepin-7-yl)sulfonyl) propanamido)butyl)(8-oxo-8-(tridecan-5-yloxy)octyl)amino)octanoate (Compound 85). UPLC- MS: m / z (ESI) for C60H108N2O9S: [M+H]+expected: 1034.59; observed: 1034.06. 'H NMR (400 MHz, CDCh) 87.50 (dq, J= 4.7, 2.3 Hz, 2H), 7.42 (s, 1H), 7.15 - 6.99 (m, 1H), 4.85 (p, J= 6.3 Hz, 1H), 4.32 (dt, J= 16.2, 5.8 Hz, 4H), 4.07 (t, J= 7.1 Hz, 2H), 3.49 (d, J= 2.3 Hz, 5H), 3.41 - 3.23 (m, 2H), 2.97 (d, J= 18.6 Hz, 7H), 2.70 (t, J= 7.2 Hz, 2H), 2.43 - 2.23 (m, 6H), 1.93 (t, J = 7.7 Hz, 2H), 1.87 - 1.44 (m, 12H), 1.30 (d, J= 35.5 Hz, 52H), 0.98 - 0.74 (m, 12H).C57H103ClN2O7SMW: 995.97Compound 86

[0412] 3-hexylnonyl 8-((4-(3-((2-chlorophenyl)sulfonyl)propanamido)butyl)(8-oxo-8- (tridecan-5-yloxy)octyl)amino)octanoate (Compound 86). UPLC-MS: m / z (ESI) for C57H103CIN2O7S: [M+H]+expected: 996.97; observed: 996.08. ' H NMR (400 MHz, CDCh) 8 8.13 (d, J= 7.8 Hz, 1H), 7.67- 7.54 (m, 2H), 7.54 - 7.43 (m, 1H), 4.85 (p, J = 6.2 Hz, 1H), 4.07 (t, J = 7.1 Hz, 2H), 3.80 (t, J = 7.1 Hz, 2H), 3.30 (q, J = 6.0 Hz, 2H), 2.98 (s, 6H), 2.68 (t, J= 7.1- 142 - LEGAL\113833648\6Hz, 2H), 2.27 (t, J= 7.4 Hz, 4H), 2.05 - 1.86 (m, 2H), 1.75 (s, 4H), 1.56 (ddt, J= 28.9, 13.6, 6.8 Hz, 13H), 1.30 (d, J= 35.6 Hz, 49H), 0.88 (td, J= 6.9, 2.4 Hz, 12H).C60H108N2O8SMW: 1017.59Compound 87

[0413] 3-hexylnonyl 8-((4-(3-((3,4-dihydro-2 / f-benzo[ / >][l,4]dioxepin-7-yl)sulfinyl) propanamido)butyl)(8-oxo-8-(tridecan-5-yloxy)octyl)amino)octanoate (Compound 87). UPLC- MS: m / z (ESI) for CeoHios^OsS: [M+H]+expected: 1018.59; observed: 1018.14. 'H NMR (400 MHz, CDC13) 87.26 (s, 1H), 7.15 (dd, J= 8.3, 2.2 Hz, 1H), 7.08 (d, J= 8.3 Hz, 1H), 6.57 (s, 1H), 4.86 (p, J= 6.3 Hz, 1H), 4.37 - 4.18 (m, 4H), 4.08 (t, J= 7.1 Hz, 2H), 3.22 (dt, J= 14.8, 7.6 Hz, 3H), 2.93 (dt, J= 13.5, 6.7 Hz, 1H), 2.66 (dt, J= 15.3, 7.5 Hz, 1H), 2.42 (d, J= 29.6 Hz, 7H), 2.33 -2.16 (m, 6H), 1.72- 1.37 (m, 33H), 1.25 (s, 34H), 0.88 (td, J=6.1, 3.4 Hz, 12H).C57H103FN2O7SMW: 979.52Compound 88

[0414] 3-hexylnonyl 8-((4-(3-((4-fluorophenyl)sulfonyl)propanamido)butyl)(8-oxo-8- (tridecan-5-yloxy)octyl)amino)octanoate (Compound 88). UPLC-MS: m / z (ESI) for C57H103FN2O7S: [M+H]+expected: 980.52; observed: 980.01. 'H NMR (400 MHz, CDCI3) 88.04 - 7.78 (m, 2H), 7.35 - 7.08 (m, 2H), 6.76 (s, 1H), 4.98 - 4.70 (m, 1H), 4.08 (t, J= 7.1 Hz, 2H), 3.59 - 3.39 (m, 2H), 3.18 (q, J= 6.2 Hz, 2H), 2.78 - 2.56 (m, 2H), 2.39 (t, J= 7.6 Hz, 6H), 2.35 - 2.20 (m, 4H), 1.60 (dt, J= 16.5, 7.0 Hz, 7H), 1.56 - 1.46 (m, 6H), 1.41 (m, 3H), 1.36 - 1.07 (m, 51H), 0.88 (tt, J= 7.1, 1.9 Hz, 12H).oC60H108N2O7SMW: 1001.59Compound 89

[0415] 3-hexylnonyl 8-((4-(3-((2,3-dihydro-177-inden-5-yl)sulfonyl)propanamido)butyl)(8- oxo-8-(tridecan-5-yloxy)octyl)amino)octanoate (Compound 89). UPLC-MS: m / z (ESI) for C60H108N2O7S: [M+H]+expected: 1002.59; observed: 1002.08. 'H NMR (400 MHz, CDCI3) 8- 143 - LEGAL\113833648\67.72 (d, J= 1.7Hz, 1H), 7.66 (dd, J= 7.8, 1.8 Hz, 1H), 7.37 (d, J = 7.8 Hz, 1H), 6.63 (s, 1H), 4.87 (p, J= 6.3 Hz, 1H), 4.08 (t, J = 7.1 Hz, 2H), 3.53 - 3.35 (m, 2H), 3.26 - 3.10 (m, 2H), 2.98 (t, J= 7.5 Hz, 4H), 2.72 - 2.57 (m, 2H), 2.37 (t, J= 7.8 Hz, 6H), 2.34 - 2.24 (m, 4H), 2.15 (p, J = 7.5 Hz, 2H), 1.76 - 1.38 (m, 23H), 1.26 (d, J= 8.2Hz, 43H), 0.88 (tt, J= 7.1, 1.9 Hz, 12H).6 '6CsyHiojNjOgSMW: 975.51Compound 90

[0416] 3-hexylnonyl 8-((4-(4,4-dioxido-2,3-dihydrobenzo[ / >][l,4]oxathiine-2-carboxamido) butyl)(8-oxo-8-(tridecan-5-yloxy)octyl)amino)octanoate (Compound 90). UPLC-MS: m / z (ESI) for C57H102N2O8S: [M+H]+expected: 976.51; observed: 976.14. 'H NMR (400 MHz, CDCh) 5 8.04 (s, 1H), 7.86 - 7.70 (m, 1H), 7.58 - 7.41 (m, 2H), 7.17 (ddd, J = 8.1, 5.2, 3.0 Hz, 1H), 5.41 -5.16 (m, 1H), 4.86 (p, J= 6.3 Hz, 1H), 4.07 (t, J= 7.1 Hz, 2H), 3.99 (dd, J= 14.1, 2.0 Hz, 1H), 3.49 (s, 2H), 2.99 (s, 7H), 2.28 (td, J= 7.4, 1.3 Hz, 4H), 1.99 (s, 2H), 1.77 (s, 6H), 1.65 - 1.42 (m, 13H), 1.30 (d, J = 39.5 Hz, 45H), 0.98 - 0.70 (m, 12H).C59H108N2O5MW: 925.52Compound 93

[0417] 3-hexylnonyl 8-((8-oxo-8-(tridecan-5-yloxy)octyl)(4-(5-phenylpentanamido)butyl) amino)octanoate (Compound 93). UPLC-MS: m / z (ESI) for C59H108N2O5: [M+H]+expected: 926.52; observed: 926.01. 'H NMR (400 MHz, CDCI3) 57.25 (d, J= 6.2 Hz, 2H), 7.23 - 7.08 (m, 3H), 4.07 (t, J= 7.1 Hz, 2H), 3.24 (d, J= 5.9 Hz, 2H), 2.62 (t, J= 7.1 Hz, 2H), 2.42 (s, 6H), 2.28 (t, J= 7.5 Hz, 4H), 2.17 (t, J = 7.0 Hz, 2H), 1.59 (ddq, J = 42.5, 21.5, 6.3 Hz, 20H), 1.28 (d, J = 20.2 Hz, 51H), 0.88 (td, J= 6.9, 2.4 Hz, 12H)C58H105N2O5SMW: 943.55Compound 94

[0418] 3-hexylnonyl 8-((8-oxo-8-(tridecan-5-yloxy)octyl)(4-(4-(phenylthio)butanamido)butyl)- 144 - LEGAL\113833648\6amino)octanoate (Compound 94). UPLC-MS: m / z (ESI) for C58H106N2O5S: [M+H]+expected: 944.55; observed: 944.01. *HNMR (400 MHz, CDCh) 87.33 (s, 1H), 7.32 - 7.23 (m, 3H), 7.15 (t, J= 7.3 Hz, 1H), 6.85 (s, 1H), 4.86 (p, J= 6.3 Hz, 1H), 4.07 (t, J= 7.1 Hz, 2H), 3.31 (q, J= 6.2 Hz, 2H), 2.97 (t, J= 7.2 Hz, 4H), 2.39 (t, J= 7.2 Hz, 2H), 2.28 (t, J= 7.4 Hz, 4H), 2.10 - 1.84 (m, 4H), 1.75 (s, 4H), 1.56 (ddt, J= 28.8, 13.7, 6.8 Hz, 17H), 1.30 (d, J= 38.4 Hz, 48H), 0.88 (td, J = 6.8, 2.5 Hz, 12H).C58H105BrN2O6MW: 1006.39Compound 95

[0419] 3-hexylnonyl 8-((4-(3-(2-bromo-4-methylphenoxy)propanamido)butyl)(8-oxo-8- (tridecan-5-yloxy)octyl)amino)octanoate (Compound 95). UPLC-MS: m / z (ESI) for C58Hio5BrN206: [M+H]+expected: 1007.39; observed: 1007.98. 'HNMR (400 MHz, CDCh) 8 7.35 (d, J= 2.1 Hz, 1H), 7.05 (dd, J= 8.4, 2.1 Hz, 1H), 6.81 (d, J= 8.2 Hz, 1H), 6.68 (s, 1H), 4.86 (t, J= 6.3 Hz, 1H), 4.25 (t, J= 5.8 Hz, 2H), 4.07 (t, J= 7.1 Hz, 2H), 3.28 (q, J= 6.4 Hz, 2H), 2.69 (t, J= 5.9 Hz, 2H), 2.37 (s, 6H), 2.32 - 2.15 (m, 7H), 1.54 (ddd, J = 29.0, 13.9, 6.8 Hz, 10H), 1.39 (s, 7H), 1.27 (d, J= 19.9 Hz, 50H), 0.88 (td, J= 6.9, 2.5 Hz, 12H).oC58H103N3O5SMW: 954.54Compound 96

[0420] 3-hexylnonyl 8-((4-(3-((4-cyanophenyl)thio)propanamido)butyl)(8-oxo-8-(tridecan-5- yloxy)octyl)amino)octanoate (Compound 96). UPLC-MS: m / z (ESI) for C58H103N3O5S: [M+H]+expected: 955.54; observed: 955.07. 'H NMR (400 MHz, CDCh) 87.62 - 7.48 (m, 2H), 7.39 - 7.30 (m, 2H), 6.57 (s, 1H), 4.86 (p, J= 6.2 Hz, 1H), 4.07 (t, J= 7.1 Hz, 2H), 3.27 (dt, J= 27.4, 6.8 Hz, 4H), 2.50 (t, J= 7.3 Hz, 2H), 2.37 (t, J= 7.8 Hz, 6H), 2.28 (t, J= 7.5 Hz, 4H), 1.55 (ddt, J= 29.0, 12.6, 6.9 Hz, 11H), 1.40 (t, J = 7.1 Hz, 5H), 1.35 - 1.12 (m, 51H), 0.88 (td, J= 7.0, 2.5 Hz, 12H).- 145 - LEGAL\113833648\6C59H108N2O9SMW: 1021.58Compound 97

[0421] 3-hexylnonyl 8-((4-(3-((3,4-dimethoxyphenyl)sulfonyl)propanamido)butyl)(8-oxo-8- (tridecan-5-yloxy)octyl)amino)octanoate (Compound 97). UPLC-MS: m / z (ESI) for C59H108N2O9S: [M+H]+expected: 1022.58; observed: 1022.01. 'H NMR (400 MHz, CDCI3) 8 7.51 (dd, J= 8.4, 2.1 Hz, 1H), 7.34 (d, J = 2.1 Hz, 1H), 6.98 (d, J= 8.4 Hz, 1H), 6.62 (s, 1H), 4.86 (p, J= 6.3 Hz, 1H), 4.08 (t, J= 7.1 Hz, 2H), 3.95 (d, J= 4.7 Hz, 6H), 3.43 (dd, J= 9.2, 6.3 Hz, 2H), 3.20 (q, J= 6.1 Hz, 2H), 2.71 - 2.52 (m, 2H), 2.39 (s, 6H), 2.28 (t, J= 7.5 Hz, 4H), 1.56 (ddt, J=28.7, 12.8, 6.7 Hz, 14H), 1.40 (d, J= 7.2 Hz, 4H), 1.39 - 1.18 (m, 49H), 0.88 (td, J = 6.8, 2.6 Hz, 12H).C58H106N2O7SMW: 975.55Compound 98

[0422] 3-hexylnonyl 8-((8-oxo-8-(tridecan-5-yloxy)octyl)(4-(3-(o-tolylsulfonyl)propanamido) butyl)amino)octanoate (Compound 98). UPLC-MS: m / z (ESI) for C58H106N2O7S: [M+H]+expected: 976.55; observed: 976.05. 'H NMR (400 MHz, CDCh) 87.97 (d, J= 7.9 Hz, 1H), 7.52 (td, J= 7.5, 1.4 Hz, 1H), 7.35 (t, J = 8.3 Hz, 2H), 6.69 (s, 1H), 4.97-4.72 (m, 1H), 4.08 (t, J = 7.1 Hz, 2H), 3.60 - 3.37 (m, 2H), 3.15 (q, J = 6.1 Hz, 2H), 2.71 (s, 3H), 2.60 (t, J = 7.6 Hz, 2H), 2.37 (t, J= 7.4 Hz, 6H), 2.28 (t, J= 7.5 Hz, 4H), 1.72 - 1.45 (m, 13H), 1.40 (t, J= 6.8 Hz, 3H), 1.28 (d, J= 24.5 Hz, 51H), 0.88 (td, J= 6.8, 2.5 Hz, 12H).C57H102BrClN2O6MW: 1026.81Compound 99

[0423] 3-hexylnonyl 8-((4-(3-(2-bromo-4-chlorophenoxy)propanamido)butyl)(8-oxo-8- (tridecan-5-yloxy)octyl)amino)octanoate (Compound 99). UPLC-MS: m / z (ESI) for C57Hio2BrClN206: [M+H]+expected: 1027.81; observed: 1027.97. 'H NMR (400 MHz, CDCh) 8 7.52 (d, J= 2.5 Hz, 1H), 7.33 - 7.22 (m, 1H), 6.85 (d, J= 8.8 Hz, 1H), 6.63 (s, 1H), 4.86 (t, J =- 146 - LEGAL\113833648\66.2 Hz, 1H), 4.28 (t, J= 5.9 Hz, 2H), 4.07 (t, J= 7.1 Hz, 2H), 3.28 (q, J= 6.3 Hz, 2H), 2.69 (t, J = 5.9 Hz, 2H), 2.48 - 2.34 (m, 6H), 2.27 (t, J= 7.6 Hz, 4H), 1.56 (ddd, J= 29.2, 15.0, 6.6 Hz, 13H), 1.39 (d, J= 7.5 Hz, 3H), 1.27 (d, J= 15.1 Hz, 51H), 0.88 (td, J= 6.9, 2.5 Hz, 12H).C58H103ClN2O6MW: 959.92Compound 100

[0424] 3-hexylnonyl 8-((4-(4-(4-chlorophenyl)-4-oxobutanamido)butyl)(8-oxo-8-(tridecan-5-yloxy)octyl)amino)octanoate (Compound 100). UPLC-MS: m / z (ESI) for C58H103CIN2O6:[M+H]+expected: 960.92; observed: 960.12. 'H NMR (400 MHz, CDCh) 58.01 - 7.80 (m, 2H), 7.43 - 7.30 (m, 2H), 6.30 (s, 1H), 4.78 (t, J= 6.2 Hz, 1H), 3.99 (t, J= 7.1 Hz, 2H), 3.33 - 3.09 (m, 4H), 2.51 (t, J= 6.6 Hz, 2H), 2.40 - 2.24 (m, 6H), 2.20 (t, J= 7.5 Hz, 4H), 1.47 (ddq, J = 27.7, 12.7, 6.3 Hz, 18H)1.29 - 1.08 (m, 50H), 0.80 (td, J= 6.9, 2.5 Hz, 15H).C58H103BrN2O6MW: 1004.37Compound 101

[0425] 3-hexylnonyl 8-((4-(4-(4-bromophenyl)-4-oxobutanamido)butyl)(8-oxo-8-(tridecan-5-yloxy)octyl)amino)octanoate (Compound 101). UPLC-MS: m / z (ESI) for CssHiosBr^CL:[M+H]+expected: 1005.37; observed: 1006.19.XH NMR (400 MHz, CDCh) 57.95 - 7.81 (m, 1H), 7.67 - 7.55 (m, 1H), 7.49 (d, J= 8.6 Hz, 1H), 7.26 (s, 2H), 6.39 (s, 1H), 4.86 (s, OH), 4.07 (t, J= 7.1 Hz, 2H), 3.41 - 3.06 (m, 3H), 2.67 (m, 2H), 2.51 - 2.33 (m, 6H), 2.28 (td, J= 7.6, 4.2 Hz, 5H), 1.81 - 1.46 (m, 14H), 1.40 (d, J= 7.1 Hz, 3H), 1.27 (d, J= 15.9 Hz, 50H), 0.88 (td, J= 7.0, 2.5 Hz, 12H).C60H110N2O7MW: 971.55Compound 102

[0426] 3-hexylnonyl 8-((4-(4-(3,4-dimethoxyphenyl)butanamido)butyl)(8-oxo-8-(tridecan-5-yloxy)octyl)amino)octanoate (Compound 102). UPLC-MS: m / z (ESI) for C60H110N2O7: [M+H]+expected: 972.55; observed: 972.17. 'H NMR (400 MHz, CDCh) 86.78 (d, J= 8.2 Hz, 1H), 6.71- 147 - LEGAL\113833648\6(d, J= 6.8 Hz, 2H), 6.01 (s, 1H), 4.86 (t, J= 6.2 Hz, 1H), 4.07 (t, J= 7.1 Hz, 2H), 3.86 (d, J= 5.1 Hz, 6H), 3.25 (q, J= 6.2 Hz, 2H), 2.59 (t, J= 7.6 Hz, 2H), 2.38 (s, 6H), 2.27 (t, J= 7.5 Hz, 4H), 2.16 (t, J= 7.5 Hz, 2H), 1.95 (d, J= 7.6 Hz, 1H), 1.55 (ddt, J= 28.7, 13.1, 6.9 Hz, 12H), 1.40 (s, 6H), 1.36 - 1.10 (m, 49H), 0.88 (td, J= 6.9, 2.5 Hz, 12H).MW: 938.48Compound 103

[0427] 3-hexylnonyl 8-((4-(3-(3-cyanophenoxy)propanamido)butyl)(8-oxo-8-(tridecan-5-yloxy)octyl)amino)octanoate (Compound 103). UPLC-MS: m / z (ESI) for C58H103N3O6: [M+H]+expected: 939.48; observed: 939.01. 'H NMR (400 MHz, CDCh) 87.36 (t, J= 7.8 Hz, 1H), 7.26 (d, J = 3.8 Hz, 1H), 7.14 (d, J= 8.5 Hz, 2H), 6.60 (s, 1H), 4.95 - 4.76 (m, 1H), 4.29 (t, J= 6.1 Hz, 2H), 4.07 (t, J= 7.1 Hz, 2H), 3.29 (q, J= 6.2 Hz, 2H), 2.64 (t, J= 6.2 Hz, 2H), 2.40 (s, 6H), 2.27 (t, J = 7.5 Hz, 4H), 1.56 (tq, J= 21.6, 6.2 Hz, 11H), 1.41 (s, 6H), 1.28 (d, J= 20.8 Hz, 49H), 0.88 (td, J= 6.9, 2.4 Hz, 12H).C61H110N2O9MW: 1015.56Compound 104

[0428] 3-hexylnonyl 8-((4-(4-oxo-4-(3,4,5-trimethoxyphenyl)butanamido)butyl)(8-oxo-8-(tridecan-5-yloxy)octyl)amino)octanoate (Compound 104). UPLC-MS: m / z (ESI) for C61H110N2O9: [M+H]+expected: 1016.56; observed: 1016.06.XH NMR (400 MHz, CDCh) 87.25 (d, J= 5.2 Hz, 2H), 6.38 (s, 1H), 5.00 - 4.69 (m, 1H), 4.07 (t, J= 7.0 Hz, 2H), 3.91 (s, 9H), 3.33 (t, J= 6.8 Hz, 2H), 3.26 (q, J= 6.3 Hz, 2H), 2.59 (t, J= 6.7 Hz, 2H), 2.39 (s, 6H), 2.27 (t, J= 7.5 Hz, 4H), 1.80 - 1.38 (m, 18H), 1.27 (d, J = 17.9 Hz, 5 OH), 0.88 (td, J= 6.9, 2.5 Hz, 12H).C56H102N2O5S2MW: 947.56Compound 105

[0429] 3-hexylnonyl 8-((8-oxo-8-(tridecan-5-yloxy)octyl)(4-(2-(phenyldisulfaneyl)acetamido) butyl)amino)octanoate (Compound 105). UPLC-MS: m / z (ESI) for C56H102N2O5S2: [M+H]+- 148 - LEGAL\113833648\6expected: 948.56; observed: 948.38. 'H NMR (400 MHz, CDCh) 7.26 (m 5H), 4.86 (t, J= 6.3 Hz, 1H), 4.07 (t, J= 7.1 Hz, 2H), 3.49 (s, 2H), 3.23 (d, J= 6.4 Hz, 1H), 2.96 (s, 6H), 2.28 (t, J = 7.2 Hz, 4H), 5 1.81 (d, J= 36.4 Hz, 8H), 1.54 (s, 18H), 1.30 (d, J= 36.8 Hz, 43H), 0.94 - 0.81 (m, 12H).C61H112N2O9MW: 1017.57Compound 106

[0430] 3-hexylnonyl 8-((4-(4-hydroxy-4-(3,4,5-trimethoxyphenyl)butanamido)butyl)(8-oxo-8-(tridecan-5-yloxy)octyl)amino)octanoate (Compound 106). UPLC-MS: m / z (ESI) for C61H112N2O9: [M+H]+expected: 1018.57; observed: 1018.23.XH NMR (400 MHz, CDCh) 87.40 (s, 1H), 6.62 (s, 2H), 4.85 (t, J= 6.2 Hz, 1H), 4.72 (dd, J= 8.5, 3.6 Hz, 1H), 4.07 (t, J= 7.1 Hz, 2H), 3.84 (dd, J= 16.3, 1.3 Hz, 9H), 3.48 - 3.23 (m, 1H), 2.94 (s, 6H), 2.5 (m, 2H), 2.28 (t, J = 7.4 Hz, 4H), 2.2-1.75(m, 4H) 1.73 (s, 5H), 1.69 - 1.41 (m, 12H), 1.30 (d, J= 37.7 Hz, 50H), 0.88 (ddd, J= 7.4, 4.8, 2.8 Hz, 12H).MW: 1001.57Compound 107

[0431] 3-hexylnonyl 8-((8-oxo-8-(tridecan-5-yloxy)octyl)(4-(4-(3,4,5-trimethoxyphenyl) butanamido)butyl)amino)octanoate (Compound 107). UPLC-MS: m / z (ESI) for C61H112N2O8:[M+H]+expected: 1002.57; observed: 1002.17.XH NMR (400 MHz, CDCh) 86.40 (s, 2H), 6.01 (s, 1H), 4.99 - 4.79 (m, 1H), 4.07 (t, J= 7.1 Hz, 2H), 3.83 (d, J= 9.2 Hz, 9H), 3.25 (q, J= 6.2 Hz, 2H), 2.67 - 2.53 (m, 2H), 2.38 (s, 6H), 2.28 (t, J= 7.5 Hz, 4H), 2.18 (t, J= 7.5 Hz, 2H), 2.04 - 1.82 (m, 2H), 1.74 - 1.47 (m, 16H), 1.42 (d, J= 14.0 Hz, 1H), 1.27 (d, J= 18.6 Hz, 49H), 0.88 (td, J = 6.9, 2.7 Hz, 12H).MW: 956.49Compound 108

[0432] 3-hexylnonyl 8-((4-(4-(4-nitrophenyl)butanamido)butyl)(8-oxo-8-(tridecan-5-- 149 - LEGAL\113833648\6yloxy)octyl)amino)octanoate (Compound 108). UPLC-MS: m / z (ESI) for C58H105N3O7 [M+H]+ expected: 957.49; observed: 957.19. 'H NMR (400 MHz, CDCh) 88.17 - 8.09 (m, 2H), 7.34 (d, J= 8.3 Hz, 2H), 4.86 (p, J= 6.4 Hz, 1H), 4.07 (t, J= 7.1 Hz, 2H), 3.25 (q, J= 6.1 Hz, 2H), 2.75 (t, J= 7.7 Hz, 2H), 2.45 (s, 6H), 2.27 (t, J= 7.5 Hz, 4H), 2.18 (t, J= 7.3 Hz, 2H), 1.98 (tt, J = 14.1, 7.6 Hz, 2H), 1.56 (ddq, J= 28.7, 13.3, 6.7 Hz, 12H), 1.45 - 1.37 (m, 2H), 1.27 (d, J= 16.3 Hz, 53H), 0.88 (td, J= 6.9, 2.5 Hz, 12H).C57H103BrN2O5MW: 976.36Compound 109

[0433] 3-hexylnonyl 8-((4-(3-(4-bromophenyl)propanamido)butyl)(8-oxo-8-(tridecan-5-yloxy)octyl)amino)octanoate (Compound 109). UPLC-MS: m / z (ESI) for CsrHiosBr^Ch [M+H]+ expected: 977.36; observed: 978.03. 'H NMR (400 MHz, CDCh) 87.41 - 7.34 (m, 2H), 7.10 - 7.01 (m, 2H), 6.28 (s, 1H), 4.86 (p, J = 6.3 Hz, 1H), 4.07 (t, J = 7.1 Hz, 2H), 3.21 (q, J = 6.2 Hz, 2H), 2.92 (t, J= 7.6 Hz, 2H), 2.41 (t, J= 7.5 Hz, 6H), 2.27 (t, J= 7.5 Hz, 4H), 1.68 - 1.37 (m, 19H), 1.28 (d, J= 20.4 Hz, 50H), 0.88 (td, J= 6.9, 2.4 Hz, 12H).C57H102BrFN2O6MW: 1010.35Comoound 110

[0434] 3-hexylnonyl 8-((4-(3-(4-bromo-2-fluorophenoxy)propanamido)butyl)(8-oxo-8-(tridecan-5-yloxy)octyl)amino)octanoate (Compound 110). UPLC-MS: m / z (ESI) for C57Hio2BrFN206 [M+H]+ expected: 1011.35; observed: 1010.11. 'HNMR (400 MHz, CDCh) 8 7.26 - 7.14 (m, 2H), 6.89 (t, J = 8.7 Hz, 1H), 6.66 (s, 1H), 4.86 (p, J = 6.3 Hz, 1H), 4.30 (t, J = 6.1 Hz, 2H), 4.07 (t, J= 7.1 Hz, 2H), 3.27 (q, J= 6.2 Hz, 2H), 2.65 (t, J= 6.1 Hz, 2H), 2.40 (s, 6H), 2.27 (t, J = 7.5 Hz, 4H), 1.56 (ddd, J= 29.0, 14.5, 7.3 Hz, 14H), 1.41 (s, 4H), 1.28 (d, J = 19.9 Hz, 49H), 0.88 (td, J= 6.9, 2.4 Hz, 12H).- 150 - LEGAL\113833648\6C56H101N3O9SMW: 99250Comoound 111

[0435] 3-hexylnonyl 8-((4-(2-((4-nitrophenyl)sulfonyl)acetamido)butyl)(8-oxo-8-(tridecan-5- yloxy)octyl)amino)octanoate (Compound 111). UPLC-MS: m / z (ESI) for C56H101N3O9S [M+H]+ expected: 993.50; observed: 993.01. 'H NMR (400 MHz, CDCh) 88.44 - 8.37 (m, 2H), 8.17- 8.10 (m, 2H), 7.62 (s, 1H), 4.87 (p, J = 6.2 Hz, 1H), 4.07 (dd, J= 14.0, 6.9 Hz, 4H), 3.25 (q, J= 6.1 Hz, 2H), 2.45 (s, 6H), 2.28 (t, J= 7.5 Hz, 4H), 1.54 (dtt, J= 28.7, 22.3, 7.1 Hz, 15H), 1.28 (d, J= 26.4 Hz, 52H), 0.88 (td, J= 6.8, 2.3 Hz, 12H).C56H101N5O5MW: 924.45Comoound 112

[0436] 3-hexylnonyl 8-((4-(2-((5-cyanopyridin-2-yl)amino)acetamido)butyl)(8-oxo-8- (tridecan-5-yloxy)octyl)amino)octanoate (Compound 112). UPLC-MS: m / z (ESI) for C56H101N5O5 [M+H]+ expected: 925.45; observed: 925.07. ' H NMR (400 MHz, CDCh) 88.37 (d, J = 2.1 Hz, 1H), 7.57 (dd, J= 8.7, 2.2 Hz, 1H), 6.86 (s, 1H), 6.50 (s, 1H), 5.77 (s, 1H), 4.86 (p, J= 6.3 Hz, 1H), 4.08 (t, J= 7.2 Hz, 4H), 3.33 - 3.26 (m, 2H), 2.38 (s, 6H), 2.28 (t, J= 7.5 Hz, 4H), 1.67 - 1.38 (m, 20H), 1.28 (d, J= 19.2 Hz, 47H), 0.88 (td, J= 6.9, 2.5 Hz, 12H).C57H102Br2N2O5MW: 1055.26Comoound 113

[0437] 3-hexylnonyl 8-((4-(3-(2,4-dibromophenyl)propanamido)butyl)(8-oxo-8-(tridecan-5- yloxy)octyl)amino)octanoate (Compound 113). UPLC-MS: m / z (ESI) for C57Hio2Br2N20s [M+H]+ expected: 1056.26; observed: 1055.93.XH NMR (400 MHz, CDCh) 87.66 (2, 1H), 7.37 - 7.27 (m, 1H), 7.16 (d, J = 8.2 Hz, 1H), 6.47 (s, 1H), 4.86 (p, J = 6.3 Hz, 1H), 4.07 (t, J = 7.1 Hz, 2H), 3.82 - 3.59 (m, 1H), 3.22 (q, J= 6.1 Hz, 2H), 3.04 (t, J= 7.6 Hz, 2H), 2.46 (dt, J= 15.2, 7.6 Hz, 6H), 2.32 - 2.24 (m, 4H), 1.67 - 1.39 (m, 20H), 1.27 (d, J= 18.7 Hz, 47H), 0.88 (td, J =- 151 - LEGAL\113833648\66.9, 2.6 Hz, 12H).oC58H106N2O8SMW: 991.55Compound 114

[0438] 3 -hexylnonyl 8-((4-(3-((4-methoxyphenyl)sulfonyl)propanamido)butyl)(8-oxo-8- (tridecan-5-yloxy)octyl)amino)octanoate (Compound 114). UPLC-MS: m / z (ESI) for C58H106N2O8S [M+H]+ expected: 992.55; observed: 992.11. 'H NMR (400 MHz, CDCh) 87.83 (d, J= 8.9 Hz, 2H), 7.01 (d, J= 8.9 Hz, 2H), 6.68 (s, 1H), 4.86 (p, J= 6.3 Hz, 1H), 4.08 (t, J = 7.1 Hz, 2H), 3.88 (s, 3H), 3.46 - 3.38 (m, 2H), 3.19 (d, J= 5.8 Hz, 2H), 2.60 (dd, J= 8.8, 6.7 Hz, 2H), 2.40 (s, 6H), 2.28 (t, J= 7.3 Hz, 4H), 1.71 - 1.38 (m, 16H), 1.36 - 1.18 (m, 51H), 0.88 (ddd, J= 7.0, 5.1, 1.8 Hz, 12H).C57H106N4O5MW: 927.50Compound 115

[0439] 3-hexylnonyl 8-((4-(3-((5-methylpyridin-2-yl)amino)propanamido)butyl)(8-oxo-8- (tridecan-5-yloxy)octyl)amino)octanoate (Compound 115). UPLC-MS: m / z (ESI) for C57H106N4O5 [M+H]+ expected: 928.50; observed: 928.33. 'H NMR (400 MHz, CDCh) 87.89 (d, J= 2.3 Hz, 1H), 7.21 (dd, J = 8.5, 2.4 Hz, 1H), 6.61 (s, 1H), 6.35 (d, J= 8.4 Hz, 1H), 4.86 (p, J= 7.4 Hz, 2H), 4.07 (t, J= 7.1 Hz, 2H), 3.63 (q, J= 6.2 Hz, 2H), 3.24 (q, J= 6.2 Hz, 2H), 2.51 -2.38 (m, 8H), 2.27 (t, J= 7.5 Hz, 4H), 2.16 (s, 3H), 1.67- 1.38 (m, 14H), 1.28 (d, J= 19.6 Hz, 53H), 0.88 (td, J= 6.9, 2.7 Hz, 12H).C57H104BrN3O5MW: 991.38Compound 116

[0440] 3-hexylnonyl 8-((4-(4-(5-bromopyridin-2-yl)butanamido)butyl)(8-oxo-8-(tridecan-5-yloxy)octyl)amino)octanoate (Compound 116). UPLC-MS: m / z (ESI) for CsrHuMBrNsOs [M+H]+ expected: 992.38; observed: 992.91. 'H NMR (400 MHz, CDC13) 88.56 (d, J= 2.4 Hz, 1H), 7.71 (dd, J = 8.3, 2.4 Hz, 1H), 7.08 (d, J= 8.3 Hz, 1H), 6.44 (s, 1H), 4.86 (p, J = 6.3 Hz,- 152 - LEGAL\113833648\61H), 4.07 (t, J= 7.1 Hz, 2H), 3.24 (p, J= 6.5 Hz, 2H), 2.79 (t, J= 7.4 Hz, 2H), 2.43 (s, 6H), 2.27 (t, J= 7.5 Hz, 4H), 2.18 (t, J = 7.2 Hz, 2H), 2.04 (p, J = 7.3 Hz, 2H), 1.55 (ddt, J = 29.0, 14.1, 6.9 Hz, 14H), 1.43 - 1.37 (m, 8H), 1.27 (d, J= 18.4 Hz, 45H), 0.87 (td, J= 6.8, 2.7 Hz, 12H).C58H102BrN3O5MW: 977.35Comnound 120

[0441] 3-hexylnonyl 8-((4-(3-(5-bromopyridin-2-yl)propanamido)butyl)(8-oxo-8-(tridecan-5-yloxy)octyl)amino)octanoate (Compound 120). UPLC-MS: m / z (ESI) for CseHn^Br sOs: [M+H]+expected: 978.35; observed: 979.26. 'H NMR (400 MHz, CDC13) 58.55 (d, J= 2.4 Hz, 1H), 7.70 (dd, J= 8.3, 2.4 Hz, 1H), 7.12 (d, J= 8.3 Hz, 1H), 6.44 (s, 1H), 4.86 (t, J = 6.2 Hz, 1H), 4.07 (t, J= 7.1 Hz, 2H), 3.21 (d, J= 6.0 Hz, 2H), 3.08 (t, J= 7.2 Hz, 2H), 2.62 (t, J= 7.2 Hz, 2H), 2.41 (s, 6H), 2.32 -2.23 (m, 4H), 1.90-1.70 (m, 4H), 1.68 - 1.37 (m, 13H), 1.27 (d, J = 19.2 Hz, 50H), 0.88 (td, J= 6.9, 2.6 Hz, 12H).oC57H102Br2N2O5MW: 1055.26Compound 121

[0442] 3-hexylnonyl 8-((4-(3-(3,4-dibromophenyl)propanamido)butyl)(8-oxo-8-(tridecan-5-yloxy)octyl)amino)octanoate (Compound 121). UPLC-MS: m / z (ESI) for C57Hio2Br2N20s: [M+H]+expected: 1056.26; observed: 1056.22.XH NMR (400 MHz, CDC13) 57.60 - 7.42 (m, 2H), 7.02 (dd, J= 8.2, 2.1 Hz, 1H), 4.86 (t, J= 6.2 Hz, 1H), 4.07 (t, J= 7.1 Hz, 2H), 3.22 (q, J = 6.3 Hz, 2H), 2.90 (t, J= 7.5 Hz, 2H), 2.40 (q, J= 9.3 Hz, 6H), 2.35 -2.16 (m, 4H), 1.72 - 1.35 (m, 22H), 1.37 - 1.19 (m, 48H), 0.88 (td, J = 6.9, 2.6 Hz, 12H).C57H103N3O6MW: 926.47Compound 122

[0443] 3-hexylnonyl 8-((4-(3-oxo-3-(phenylamino)propanamido)butyl)(8-oxo-8-(tridecan-5-yloxy)octyl)amino)octanoate (Compound 122). UPLC-MS: m / z (ESI) for C57H103N3O6: [M+H]+- 153 - LEGAL\113833648\6expected: 927.47; observed: 927.01. 'H NMR (400 MHz, CDC13) 89.69 (s, 1H), 7.74 (d, J= 5.4 Hz, 1H), 7.56 (d, J= 7.9 Hz, 2H), 7.32 (d, J= 8.1 Hz, 2H), 7.09 (t, J= 7.4 Hz, 1H), 4.86 (t, J = 6.2 Hz, 1H), 4.08 (t, J= 7.1 Hz, 2H), 3.28 (d, J= 7.8 Hz, 4H), 2.41 (q, J= 6.5 Hz, 6H), 2.28 (t, J = 7.5 Hz, 4H), 1.71 - 1.36 (m, 16H), 1.27 (d, J= 19.7 Hz, 51H), 0.88 (td, J= 6.9, 2.6 Hz, 12H).C60H110N2O8SMW: 1019.61Compound 123

[0444] 3-hexylnonyl 8-((8-oxo-8-(tridecan-5-yloxy)octyl)(4-(3-((3,4,5-trimethoxyphenyl)thio) propanamido)butyl)amino)octanoate (Compound 123). UPLC-MS: m / z (ESI) for C60H110N2O8S:[M+H]+expected: 1020.61; observed: 1020.17. 'HNMR (400 MHz, CDCh) 87.77 (s, 1H), 7.26 (s, 1H), 6.79 (s, 1H), 5.45 - 5.30 (m, 1H), 4.58 (t, J= 7.1 Hz, 2H), 4.43 -4.28 (m, 13H), 3.88 -3.67 (m, 4H), 2.97 (d, J= 7.4 Hz, 2H), 2.96 - 2.84 (m, 6H), 2.79 (t, J= 7.5 Hz, 4H), 2.27 - 1.85 (m, 16H), 1.79 (d, J= 21.8 Hz, 53H), 1.39 (td, J= 6.8, 2.7 Hz, 12H).C58H103FN2O6MW: 943.47Compound 124

[0445] 3-hexylnonyl 8-((4-(4-(4-fluorophenyl)-4-oxobutanamido)butyl)(8-oxo-8-(tridecan-5-yloxy)octyl)amino)octanoate (Compound 124). UPLC-MS: m / z (ESI) for C58H103FN2O6 [M+H]+ expected: 943.47; observed: 943.98. 1H NMR (400 MHz, CDCI3) 88.05 - 7.97 (m, 2H), 7.13 (t, J = 8.6 Hz, 2H), 4.86 (p, J = 6.3 Hz, 1H), 4.07 (t, J= 7.1 Hz, 2H), 3.32 (t, J= 6.6 Hz, 2H), 3.29 - 3.21 (m, 2H), 2.60 (t, J= 6.6 Hz, 2H), 2.40 (s, 6H), 2.27 (t, J= 7.5 Hz, 4H), 1.63 -1.46 (m, 19H), 1.41 (s, 1H), 1.27 (d, J= 17.4 Hz, 47H), 0.88 (td, J= 6.9, 2.6 Hz, 12H).C56H102N2O7SMW: 947.50Compound 125

[0446] 3-hexylnonyl 8-((8-oxo-8-(tridecan-5-yloxy)octyl)(4-(2-(phenylsulfonyl)acetamido) butyl)amino)octanoate (Compound 125). UPLC-MS: m / z (ESI) for C56H102N2O7S [M+H]+ expected: 947.5; observed: 947.98. 'H NMR (400 MHz, CDC13) 87.92 (d, J= 7.7 Hz, 2H), 7.69- 154 - LEGAL\113833648\6(t, J= 7.4 Hz, 1H), 7.58 (t, J= 7.7 Hz, 2H), 4.87 (p, J= 6.3 Hz, 1H), 4.08 (t, J= 7.1 Hz, 2H), 3.99 (s, 2H), 3.28 (s, 2H), 2.40 (s, 6H), 2.32 - 2.22 (m, 4H), 1.65 - 1.46 (m, 17H), 1.41 (s, 1H), 1.36 - 1.16 (m, 49H), 0.88 (td, J = 6.8, 2.6 Hz, 12H).o< -.... X..11... Q......1........611C59H109N2O7PMW: 989.50Compound 126

[0447] 3 -hexylnonyl 8-((4-(3-(ethoxy(phenyl)phosphoryl)propanamido)butyl)(8-oxo-8- (tridecan-5-yloxy)octyl)amino)octanoate (Compound 126). UPLC-MS: m / z (ESI) for C59H109N2O7P: [M+H]+expected: 990.50; observed: 990.12. 'H NMR (400 MHz, CDCh) 87.78 (ddt, J= 11.7, 6.8, 1.5 Hz, 2H), 7.62 - 7.39 (m, 3H), 7.00 (s, 1H), 4.86 (p, J = 6.3 Hz, 1H), 4.07 (td, J= 7.2, 2.2 Hz, 3H), 3.87 (dt, J= 10.1, 7.3 Hz, 1H), 3.25 (q, J= 6.3 Hz, 2H), 2.94 (s, 6H), 2.58 - 2.34 (m, 1H), 2.28 (td, J= 7.5, 1.4 Hz, 5H), 1.98 - 1.41 (m, 23H), 1.44 - 1.09 (m, 49H), 0.88 (tt, J= 7.1, 1.9 Hz, 12H).C57H102BrFN2O6MW: 1010.35Compound 127

[0448] 3 -hexylnonyl 8-((4-(3-(2-bromo-4-fluorophenoxy)propanamido)butyl)(8-oxo-8- (tridecan-5-yloxy)octyl)amino)octanoate (Compound 127). UPLC-MS: m / z (ESI) for C57Hio2BrFN206: [M+H]+expected: 1011.35; observed: 1010.11. 'HNMR (400 MHz, CDCh) 8 7.32 - 7.22 (m, 1H), 6.99 (ddd, J= 9.1, 7.7, 3.0 Hz, 1H), 6.88 (dd, J= 9.1, 4.7 Hz, 1H), 6.65 (s, 1H), 4.93 - 4.79 (m, 1H), 4.26 (t, J= 5.9 Hz, 2H), 4.07 (t, J= 7.1 Hz, 2H), 3.28 (q, J= 6.3 Hz, 2H), 2.68 (t, J= 5.9 Hz, 2H), 2.38 (s, 6H), 2.27 (t, J= 7.5 Hz, 4H), 1.72 - 1.36 (m, 18H), 1.36 - 1.13 (m, 49H), 0.88 (td, J= 6.9, 2.7 Hz, 12H).C60H110N2O10SMW: 1051.60Compound 128

[0449] 3-hexylnonyl 8-((8-oxo-8-(tridecan-5-yloxy)octyl)(4-(3-((3,4,5-trimethoxyphenyl) sulfonyl)propanamido)butyl)amino)octanoate (Compound 128). UPLC-MS: m / z (ESI) for- 155 - LEGAL\113833648\6C60H110N2O10S: [M+H]+expected: 1052.60; observed: 1052.01. 'H NMR (400 MHz, CDC13) 8 7.13 (d, J= 1.9 Hz, 2H), 4.84 (s, 1H), 4.06 (d, J= 1.8 Hz, 2H), 3.95 - 3.71 (m, 10H), 3.50 (t, J = 7.4 Hz, 2H), 3.28 (d, J= 6.3 Hz, 2H), 2.99 (s, 6H), 2.71 (t, J= 7.3 Hz, 1H), 2.26 (t, J= 7.4 Hz, 4H), 1.90 (s, 3H), 1.83 - 1.44 (m, 13H), 1.29 (d, J= 36.9 Hz, 51H), 0.99 - 0.70 (m, 12H).C57H102N2O7MW: 927.45Comoound 129

[0450] 3-hexylnonyl 8-((4-(3-oxo-3-phenoxypropanamido)butyl)(8-oxo-8-(tridecan-5-yloxy)octyl)amino)octanoate (Compound 129). UPLC-MS: m / z (ESI) for C C57H102N2O7:[M+H]+expected: 928.45; observed: 928.09. ' H NMR (400 MHz, CDCI3) 87.94 - 7.71 (m, 1H), 7.44 - 7.31 (m, 2H), 7.24 (d, J= 13.2 Hz, 1H), 7.12 (dd, J= 7.5, 1.7 Hz, 2H), 4.97-4.71 (m, 1H), 4.07 (t, J= 7.1 Hz, 2H), 3.62 (s, 2H), 3.36 (dq, J= 6.4, 3.4 Hz, 2H), 2.87 (d, J= 24.4 Hz, 6H), 2.27 (tt, J= 7.6, 1.5 Hz, 4H), 1.83 (s, 2H), 1.72 - 1.35 (m, 16H), 1.27 (d, J= 18.0 Hz, 52H), 0.88 (td, 7 = 6.2, 3.6 Hz, 12H).6C59H106BrN3O6SMW: 1065.48Compound 130

[0451] 3-hexylnonyl 8-((4-(2-acetamido-3-((4-bromophenyl)thio)propanamido)butyl)(8-oxo-8-(tridecan-5-yloxy)octyl)amino)octanoate (Compound 130). UPLC-MS: m / z (ESI) for C59Hio6BrN306S: [M+H]+expected: 1066.48; observed: 1065.48.XH NMR (400 MHz, CDCh) 8 7.42 (d, J= 8.5 Hz, 2H), 7.37 - 7.28 (m, 2H), 6.78 (s, 1H), 6.43 (s, 1H), 4.86 (p, J= 6.3 Hz, 1H), 4.46 (d, J= 6.8 Hz, 1H), 4.07 (t, J= 7.1 Hz, 2H), 3.40 - 3.06 (m, 4H), 2.39 (s, 6H), 2.28 (t, J = 7.5 Hz, 4H), 1.98 (s, 2H), 1.77- 1.37 (m, 16H), 1.28 (d, J= 19.8 Hz, 51H), 0.88 (td, J= 6.9, 2.6 Hz, 12H).- 156 - LEGAL\113833648\6MW: 997.57Comnound 131

[0452] 3-hexylnonyl 8-((8-oxo-8-(tridecan-5-yloxy)octyl)(4-(3-((l-phenyl-l / f-tetrazol-5-yl)thio)propanamido)butyl)amino)octanoate. (Compound 131). UPLC-MS: m / z (ESI) for C58H104N6O5S: [M+H]+expected: 998.57; observed: 998.00. 'H NMR (400 MHz, CDCh) 87.69 - 7.46 (m, 5H), 6.57 (s, 1H), 4.86 (p, J= 6.3 Hz, 1H), 4.07 (t, J= 7.1 Hz, 2H), 3.64 (t, J= 6.7 Hz, 2H), 3.26 (q, J= 6.2 Hz, 2H), 2.81 (t, J= 6.6 Hz, 2H), 2.38 (s, 5H), 2.27 (t, J= 7.5 Hz, 4H), 1.73 - 1.36 (m, 18H), 1.26 (d, J= 11.4 Hz, 51H), 0.88 (td, J= 6.9, 2.5 Hz, 12H).C57H103N3O5MW: 926.47Comoound 132

[0453] 3-hexylnonyl 8-((4-(4-oxo-4-(pyridin-2-yl)butanamido)butyl)(8-oxo-8-(tridecan-5-yloxy)octyl)amino)octanoate (Compound 132). UPLC-MS: m / z (ESI) for C57H103N3O6 [M+H]+ expected: 926.47; observed: 927.09. 'H NMR (400 MHz, CDCh) 88.56 (d, J= 4.9 Hz, 1H), 7.80 (t, J= 7.9 Hz, 1H), 7.43 -7.29 (m, 2H), 4.86 (p, J= 6.3 Hz, 1H), 4.07 (t, J= 7.1 Hz, 2H), 3.21 (dt, J = 25.9, 8.1 Hz, 2H), 2.73 (ddd, J= 24.7, 12.2, 7.4 Hz, 2H), 2.56 (ddd, J= 17.2, 9.1, 5.5 Hz, 2H), 2.29 (dt, J = 15.3, 6.0 Hz, 10H), 1.55 (ddt, J= 28.8, 12.7, 6.7 Hz, 13H), 1.27 (d, J= 18.4 Hz, 54H), 0.88 (h, J= 2.6 Hz, 12H).C57H105N3O5MW: 912.48Compound 133

[0454] 3-hexylnonyl 8-((8-oxo-8-(tridecan-5-yloxy)octyl)(4-(4-(pyridin-2-yl)butanamido) butyl)amino)octanoate (Compound 133). UPLC-MS: m / z (ESI) for C57H105N3O5 [M+H]+ expected: 912.48; observed: 913.15. 'H NMR (400 MHz, CDCh) 88.50 (d, J= 4.9 Hz, 1H), 7.60 (t, J= 7.7 Hz, 1H), 7.20 - 7.08 (m, 2H), 6.60 (s, 1H), 4.86 (t, J= 6.3 Hz, 1H), 4.07 (t, J= 7.1 Hz, 2H), 3.26 (q, J= 6.1 Hz, 2H), 2.83 (t, J= 7.3 Hz, 2H), 2.39 (s, 6H), 2.27 (t, J= 7.5 Hz, 4H), 2.19 (t, J= 7.3 Hz, 2H), 2.06 (p, J = 7.2 Hz, 2H), 1.68 - 1.47 (m, 12H), 1.40 (s, 8H), 1.27 (d, J= 19.6- 157 - LEGAL\113833648\6Hz, 47H), 0.87 (h, J = 2.7 Hz, 12H).MW; 929.49Compound 134

[0455] 3-hexylnonyl 8-((4-(4-(4-fluorophenyl)butanamido)butyl)(8-oxo-8-(tridecan-5-yloxy)octyl)amino)octanoate (Compound 134). UPLC-MS: m / z (ESI) for C58H105FN2O5 [M+H]+ expected: 929.49; observed: 930.06. 'H NMR (400 MHz, CDCh) 87.12 (dd, J= 8.4, 5.3 Hz, 2H), 6.95 (t, J= 8.7 Hz, 2H), 6.16 (s, 1H), 4.86 (t, J= 6.3 Hz, 1H), 4.07 (t, J= 7.1 Hz, 2H), 3.24 (q, J= 6.1 Hz, 2H), 2.62 (t, J= 7.6 Hz, 2H), 2.39 (q, J= 8.4 Hz, 6H), 2.27 (t, J= 7.6 Hz, 4H), 2.14 (t, J= 7.5 Hz, 2H), 1.94 (p, J= 7.6 Hz, 2H), 1.78 (s, 2H), 1.67- 1.46 (m, 11H), 1.39 (d, J= 7.5 Hz, 1H), 1.28 (d, J= 21.6 Hz, 53H), 0.92 - 0.83 (m, 12H).C56H103N3O7SMW: 962.51Compound 135

[0456] 3-hexylnonyl 8-((8-oxo-8-(tridecan-5-yloxy)octyl)(4-(2-(phenylsulfonamido) acetamido)butyl)amino)octanoate (Compound 135). UPLC-MS: m / z (ESI) for C56H103N3O7S: [M+H]+expected: 963.51; observed: 963.09. 'H NMR (400 MHz, CDCh) 87.97 - 7.76 (m, 2H), 7.68 - 7.43 (m, 3H), 6.99 (d, J= 5.6 Hz, 1H), 4.86 (p, J= 6.3 Hz, 1H), 4.07 (t, J= 7.1 Hz, 2H), 3.55 (s, 2H), 3.21 (d, J= 6.1 Hz, 1H), 2.36 (t, J= 7.6 Hz, 6H), 2.28 (t, J= 7.5 Hz, 4H), 1.85 - 1.36 (m, 20H), 1.35 - 1.13 (m, 48H), 0.88 (td, J= 6.9, 2.7 Hz, 12H).oC69H115ClF2N2O7SMW; 1190.19Compound 136

[0457] 3-hexylnonyl 8-((4-(3-((ls,4r)-4-((4-chlorophenyl)sulfonyl)-4-(2,5-difluorophenyl) cyclohexyl)propanamido)butyl)(8-oxo-8-(tridecan-5-yloxy)octyl)amino)octanoate (Compound 136). UPLC-MS: m / z (ESI) for C69H115CIF2N2O7S: [M+H]+expected: 1191.19; observed:1190.38.XH NMR (400 MHz, CDCh) 87.33 (q, J= 8.7 Hz, 4H), 7.04 (dddd, J= 12.5, 9.1, 6.4, 3.3 Hz, 2H), 6.82 (ddd, J= 12.1, 8.9, 4.7 Hz, 1H), 4.86 (t, J = 6.2 Hz, 1H), 4.07 (t, J= 7.1 Hz, - 158 - LEGAL\113833648\62H), 3.25 (d, J= 6.0 Hz, 2H), 2.51 - 2.31 (m, 9H), 2.28 (t, J= 7.5 Hz, 4H), 2.17 (t, J= 7.7 Hz, 2H), 1.81 (d, J= 7.8 Hz, 2H), 1.64 (m, 14H), 1.55 - 1.37 (m, 6H), 1.28 (d, J= 23.4 Hz, 54H), 0.88 (td, J= 6.9, 2.7 Hz, 12H).oOC57H103N3O6MW: 926.47Compound 137

[0458] 3-hexylnonyl 8-((4-(2-benzamidoacetamido)butyl)(8-oxo-8-(tridecan-5-yloxy)octyl) amino)octanoate (Compound 137). UPLC-MS: m / z (ESI) for C57H103N3O6: [M+H]+expected: 927.47; observed: 927.23. 'H NMR (400 MHz, CDCh) 87.83 (dd, J= 7.0, 1.8 Hz, 2H), 7.58 -7.37 (m, 3H), 7.02 (d, J = 6.0 Hz, 2H), 5.00 -4.78 (m, 1H), 4.07 (dd, J = 7.9, 6.0 Hz, 4H), 3.30 (q, J= 6.3 Hz, 2H), 2.39 (q, J= 7.3 Hz, 6H), 2.28 (t, J= 7.5 Hz, 4H), 1.74 - 1.46 (m, 14H), 1.41 (t, J = 7.2 Hz, 3H), 1.36 - 1.12 (m, 49H), 0.88 (td, J = 6.9, 2.7 Hz, 12H).! 9oo HC58H105N3O6MW: 940.49Compound 138

[0459] 3-hexylnonyl 8-((4-(2-(JV-methylbenzamido)acetamido)butyl)(8-oxo-8-(tridecan-5-yloxy)octyl)amino)octanoate (Compound 138). UPLC-MS: m / z (ESI) for CssHiosNsOe: [M+H]+expected: 941.49; observed: 941.31. 'H NMR (400 MHz, CDCh) 87.60 - 7.32 (m, 5H), 4.86 (t, J = 6.2 Hz, 1H), 4.26 - 3.97 (m, 3H), 3.29 (d, J= 6.4 Hz, 2H), 3.09 (s, 3H), 2.40 (s, 6H), 2.32 -2.16 (m, 4H), 1.72 - 1.38 (m, 19H), 1.26 (d, J= 9.9 Hz, 50H), 0.88 (td, J= 6.9, 2.7 Hz, 12H).-O'HO.C58H10SN2O6MW: 927.49Comnound 178

[0460] 3-hexylnonyl 8-((4-(4-(4-hydroxyphenyl)butanamido)butyl)(8-oxo-8-(tridecan-5-yloxy)octyl)amino)octanoate (Compound 178). UPLC-MS: m / z (ESI) for C58H106N2O6 [M+H]+ expected: 927.49; observed: 928.46. 'H NMR (400 MHz, CDCh) 87.00 (d, J= 8.2 Hz, 2H), 6.78 - 6.72 (m, 2H), 6.21 (t, J= 5.5 Hz, 1H), 4.87 (p, J= 6.2 Hz, 1H), 4.08 (t, J= 7.1 Hz, 2H), 3.20 (q, J= 6.0 Hz, 2H), 2.57 (t, J= 7.2 Hz, 2H), 2.41 (s, 6H), 2.28 (t, J= 7.5 Hz, 4H), 2.11 (t, J= 7.4 Hz,- 159 - LEGAL\113833648\62H), 1.93 (p, J = 7.3 Hz, 2H), 1.67- 1.36 (m, 20H), 1.28 (d, J = 19.1 Hz, 47H), 0.88 (tt, J= 7.1, 1.9 Hz, 12H).HC56H103N3O5SMW: 930.52Compound 179

[0461] 3-hexylnonyl 8-((8-oxo-8-(tridecan-5-yloxy)octyl)(4-(2-((pyridin-4-ylmethyl)thio) acetamido)butyl)amino)octanoate (Compound 179). UPLC-MS: m / z (ESI) for C56H103N3O5S [M+H]+ expected: 930.52; observed: 931.36. 'H NMR (400 MHz, CDCh) 58.59 - 8.53 (m, 2H), 7.26 - 7.22 (m, 2H), 6.91 (s, 1H), 4.86 (p, J= 6.3 Hz, 1H), 4.07 (t, J= 7.1 Hz, 2H), 3.71 (s, 2H), 3.22 (q, J= 6.3 Hz, 2H), 3.07 (s, 2H), 2.43 (s, 6H), 2.27 (t, J= 7.5 Hz, 4H), 1.67 - 1.37 (m, 18H), 1.27 (d, J = 20.0 Hz, 49H), 0.87 (td, J = 6.9, 2.7 Hz, 12H).C60H109N3O6MW: 968.55Compound 180

[0462] 3-hexylnonyl 8-((4-(2-acetamido-4-phenylbutanamido)butyl)(8-oxo-8-(tridecan-5- yloxy)octyl)amino)octanoate (Compound 180). UPLC-MS: m / z (ESI) for C60H109N3O6 [M+H]+ expected: 968.55; observed: 969.51. 'H NMR (400 MHz, CDCh) 57.27 (s, 2H), 7.20 - 7.14 (m, 3H), 4.86 (p, J= 6.3 Hz, 1H), 4.39 (q, J= 7.2 Hz, 1H), 4.07 (t, J= 7.1 Hz, 2H), 3.25 (s, 2H), 2.66 (pd, J= 14.1, 6.3 Hz, 2H), 2.40 (s, 6H), 2.27 (t, J= 7.5 Hz, 4H), 2.15 (ddt, J= 12.8, 9.3, 6.3 Hz, 1H), 1.98 (s, 4H), 1.66 - 1.46 (m, 14H), 1.41 (s, 4H), 1.27 (d, J= 12.8 Hz, 49H), 0.88 (td, J= 6.9, 2.6 Hz, 12H).C58H103N5O5SMW: 982.55Compound 186

[0463] 3-hexylnonyl 8-((8-oxo-8-(tridecan-5-yloxy)octyl)(4-(2-((4-phenyl-4H-l,2,4-triazol-3- yl)thio)acetamido)butyl)amino)octanoate (Compound 186). UPLC-MS: m / z (ESI) for C58H103N5O5S [M+H]+ expected: 982.55; observed: 983.69. 'H NMR (400 MHz, CDCh) 88.29- 160 - LEGAL\113833648\6(s, 1H), 7.82 (s, 1H), 7.59 - 7.49 (m, 3H), 7.40 - 7.33 (m, 2H), 4.86 (p, J = 6.3 Hz, 1H), 4.07 (t, J = 7.1 Hz, 2H), 3.82 (s, 2H), 3.26 (q, J= 6.5 Hz, 2H), 2.38 (s, 6H), 2.27 (t, J= 7.5 Hz, 4H), 1.71 - 1.46 (m, 12H), 1.42 - 1.37 (m, 7H), 1.28 (d, J= 20.5 Hz, 48H), 0.88 (td, J= 7.0, 2.7 Hz, 12H).HMW: 873.41Compound 187

[0464] 3-hexylnonyl 8-((4-(2-(lH-imidazol-l-yl)acetamido)butyl)(8-oxo-8-(tridecan-5-yloxy) octyl)amino)octanoate (Compound 187). UPLC-MS: m / z (ESI) for C53H100N4O5 [M+H]+ expected: 873.41; observed: 874.27. 'H NMR (400 MHz, CDCh) 87.53 (s, 1H), 7.14 (s, 1H), 6.97 (d, J= 1.4 Hz, 1H), 4.86 (p, J= 6.3 Hz, 1H), 4.63 (s, 2H), 4.07 (t, J= 7.1 Hz, 2H), 3.25 (q, J = 6.4 Hz, 2H), 2.41 (s, 6H), 2.28 (t, J = 7.5 Hz, 4H), 1.67- 1.35 (m, 18H), 1.35 - 1.17 (m, 49H), 0.88 (td, J= 7.1, 2.7 Hz, 12H).CggHio / BrNzOgMW: 1020.42Compound 218

[0465] 3-hexylnonyl 8-((4-(4-(4-bromo-3-methoxyphenyl)butanamido)butyl)(8-oxo-8- (tridecan-5-yloxy)octyl)amino)octanoate (Compound 218). UPLC-MS: m / z (ESI) for C59H107BrN2O6[M+H]+ expected: 1020.42; observed: 1022.24.XH NMR (400 MHz, CDCh) 8 7.40 (d, J= 8.0 Hz, 1H), 6.73 (d, J= 1.9 Hz, 1H), 6.65 (dd, J= 8.0, 1.9 Hz, 1H), 6.24 (s, 1H), 4.86 (p, J= 6.3 Hz, 1H), 4.07 (t, J= 7.1 Hz, 2H), 3.87 (s, 3H), 3.24 (q, J= 6.3 Hz, 2H), 2.61 (t, J = 7.6 Hz, 2H), 2.44 (dt, J= 15.8, 7.0 Hz, 6H), 2.27 (t, J= 7.5 Hz, 4H), 2.16 (t, J= 7.4 Hz, 2H), 2.01 - 1.89 (m, 2H), 1.67 - 1.36 (m, 18H), 1.27 (d, J= 20.2 Hz, 49H), 0.87 (td, J= 6.9, 2.7 Hz, 12H).CgyH-iosBrNjOgMW: 1024.36Compound 249

[0466] 3-hexylnonyl 8-((4-(2-(2-bromo-3-hydroxy-4-methoxyphenoxy)acetamido)butyl)(8- oxo-8-(tridecan-5-yloxy)octyl)amino)octanoate (Compound 249). UPLC-MS: m / z (ESI) for- 161 - LEGAL\113833648\6C57Hio3BrN208 [M+H]+ expected: 1025.36; observed: 1025.9. 'H NMR (400 MHz, CDC13) 8 6.98 (t, J= 5.5 Hz, 1H), 6.77 (d, J= 8.9 Hz, 1H), 6.38 (d, J= 8.9 Hz, 1H), 4.86 (p, J= 6.3 Hz, 1H), 4.47 (d, J= 6.0 Hz, 2H), 4.07 (t, J= 7.1 Hz, 2H), 3.88 (d, J = 3.4 Hz, 3H), 3.38 (q, J = 6.5 Hz, 2H), 2.46 - 2.35 (m, 6H), 2.27 (t, J= 7.5 Hz, 4H), 1.66 - 1.46 (m, 11H), 1.40 (s, 7H), 1.27 (d, J = 20.5 Hz, 49H), 0.88 (td, J= 6.9, 2.6 Hz, 12H).CseH-iipCINgOgMW: 972.92Comnound 250

[0467] 3 -hexylnonyl 8-((4-(3-(2-chloro-4-cyanophenoxy)propanamido)butyl)(8-oxo-8- (tridecan-5-yloxy)octyl)amino)octanoate (Compound 250). UPLC-MS: m / z (ESI) for C58H102CIN3O6 [M+H]+ expected: 973.92; observed: 973.03. 'H NMR (400 MHz, CDCI3) 87.64 (d, J = 2.1 Hz, 1H), 7.54 (dd, J = 8.6, 2.1 Hz, 1H), 7.02 (d, J = 8.6 Hz, 1H), 6.69 (d, J = 5.5 Hz, 1H), 4.86 (p, J= 6.3 Hz, 1H), 4.39 (t, J= 6.1 Hz, 2H), 4.07 (t, J= 7.1 Hz, 2H), 3.28 (q, J= 6.2 Hz, 2H), 2.70 (t, J= 6.0 Hz, 2H), 2.39 (q, J= 7.7 Hz, 6H), 2.27 (t, J= 7.5 Hz, 4H), 1.71-1.39 (m, 18H), 1.27 (d, J= 16.0 Hz, 49H), 0.87 (td, J= 6.9, 2.5 Hz, 12H).B-18. Monoamine synthesis - Scheme 11:Scheme 11.o- J;XW9 r.. v M ~ J*LHC!. H.: N OXXV!EDC-HC!DMAPEt;iNDCMC49H8ONZO0MW: 793.19XXXVHStep 1: Preparation of benzyl 8-[[8-(l-butylnonoxy)-8-oxo-octyl]-[4-(3-phenoxypropanoyl amino)butyl]amino]octanoate (Intermediate XXXVII; Compound 139)

[0468] Following the general procedure for headgroup amide coupling, benzyl 8-[4-- 162 - LEGAL\113833648\6aminobutyl-[8-(l-butylnonoxy)-8-oxo-octyl]amino]octanoate (Intermediate XXVI; 13 g, 19.1 mmol, 1 eq, HC1 salt) was reacted with 3-phenoxypropanoic acid (3.17 g, 19.1 mmol, 1 eq) followed by an aqueous workup. Purification by silica gel chromatography eluting with 100% hexanes going to 10% hexanes / 90% EtOAc gave 11.3 g (14.25 mmol, 74.7% yield) of the title compound as a light yellow oil. UPLC-MS: m / z (ESI) for C51H84N2O7S [M+H]+expected:793.61; observed: 793.7; 'H-NMR (400MHz, CDCh, d): 7.38-7.26 (m, 7H), 6.90 (m, 3H), 6.57 (br s, 1H); 5.12 (s, 2H), 4.89 (m, 1H), 4.26 (m, 2H), 3.29 (m, 2H), 2.65 (m, 2H), 2.42-2.34 (m, 8H), 2.28 (m, 2H), 1.53 (m, 4H), 1.36 (m, 8H), 1.29 (m, 4H), 1.26 (m, 28H), 0.88 (m, 6H).Preparation of 3 -hexylnonyl 8-((8-oxo-8-(pentadecan-6-yloxy)octyl)(4-(3 -(phenylsulfonyl) propanamido)butyl)amino)octanoate (Compound 210)DXXiXDMAP „ J<:' i: o r o rDCM o A o N11■' ° ■'C58H10SN2O7SMW: 989.58Compound 210

[0469] To a solution of 8-((8-((3-hexylnonyl)oxy)-8-oxooctyl)(4-(3 -(phenylsulfonyl) propanamido)butyl)amino)octanoic acid (Compound XXIX, 110 mg, 0.14 mmol, 1 equiv.) in DCM (5 mL) at 0 °C was added DCC (43.7 mg, 0.21 mmol, 1.5 equiv.), DMAP (4.3 mg, 0.035 mmol, 0.25 equiv. ), and pentadecan-6-ol (48.4 mg, 0.21 mmol, 1.5 equiv.). The resulting mixture was then allowed to warm room temperature over 16 hours. The mixture was diluted with DCM and washed with a saturated sodium bicarbonate solution, dried over anhydrous MgSCE, fdtered, and the filtrate concentrated in vacuo. The crude material was purified by silica gel chromatography, eluting with 0%-l 5% MeOH in DCM to give 97.3 mg (0.1 mmol, 70% yield) of the title compound. UPLC-MS: m / z (ESI) for C59H108N2O7S [M+H]+ expected: 990.58; observed: 990.49; 'H NMR (400 MHz, CDCh) 87.94 - 7.88 (m, 2H), 7.70 - 7.62 (m, 1H), 7.57 (t, J= 7.6 Hz, 2H), 6.76 (s, 1H), 4.86 (p, J = 6.3 Hz, 1H), 4.08 (t, J = 7.1 Hz, 2H), 3.51 - 3.39 (m, 2H), 3.18 (q, J = 6.1 Hz, 2H), 2.61 (dd, J = 9.1, 6.5 Hz, 2H), 2.49 -2.36 (m, 6H), 2.28 (td, J = 7.5, 1.8 Hz, 4H), 1.76 - 1.38 (m, 16H), 1.36 - 1.20 (m, 55H), 0.87 (dt, J = 7.0, 3.3 Hz, 12H).

[0470] Following the same procedure as for Compound 210 the following compounds were prepared from 8-((8-((3-hexylnonyl)oxy)-8-oxooctyl)(4-(3 -(phenylsulfonyl)- 163 - LEGAL\113833648\6propanamido)butyl)amino)octanoic acid (Compound XXIX) and the corresponding alcohol starting materials.oC6OH1WN20-, SMW: 1003.61Compound 143

[0471] 2-hexyldecyl -((8-((3-hexylnonyl)oxy)-8-oxooctyl)(4-(3 (phenylsulfonyl)propanamido) butyl)amino)octanoate (Compound 143). UPLC-MS: m / z (ESI) for C60H110N2O7S: [M+H]+expected: 1004.61; observed: 1004.33 'H-NMR (400MHz, CDCh, d): 88.01 - 7.81 (m, 2H), 7.66 (t, J = 7.4 Hz, 1H), 7.57 (t, J = 7.6 Hz, 2H), 6.77 (s, 1H), 4.08 (t, J = 7.1 Hz, 2H), 3.96 (d, J = 5.8 Hz, 2H), 3.56 - 3.37 (m, 2H), 3.19 (d, J = 5.9 Hz, 2H), 2.73 - 2.50 (m, 2H), 2.42 (s, 6H), 2.29 (td, J = 7.5, 5.7 Hz, 4H), 1.72 - 1.46-1.42 b(m, 24H)), 1.36 - 1.10 (m, 48H), 0.88 (t, J = 6.7 Hz, 12H).o'* bMW: 1031.66Comnound 211

[0472] 3 -hexylnonyl 8-((8-(octadecan-9-yloxy)-8-oxooctyl)(4-(3 -(phenylsulfonyl) propan amido)butyl)amino)octanoate (Compound 211). UPLC-MS: m / z (ESI) for C62H114N2O7S [M+H]+ expected: 1032.66; observed: 1032.68; 'H NMR (400 MHz, CDCh) 87.95 - 7.88 (m, 2H), 7.71 - 7.62 (m, 1H), 7.57 (t, J= 7.7 Hz, 2H), 6.76 (s, 1H), 4.86 (p, J= 6.3 Hz, 1H), 4.08 (t, J = 7.1 Hz, 2H), 3.52 - 3.40 (m, 2H), 3.18 (q, J= 6.1 Hz, 2H), 2.61 (dd, J= 8.9, 6.7 Hz, 2H), 2.40 (t, J= 7.2 Hz, 6H), 2.28 (td, J= 7.5, 2.2 Hz, 4H), 1.75 - 1.38 (m, 16H), 1.36 - 1.18 (m, 61H), 0.88 (td, J= 6.9, 2.2 Hz, 12H).C55H100M2O7SMW: 933.47Compound 212

[0473] 3 -hexylnonyl 8-((8-oxo-8-(undecan-4-yloxy)octyl)(4-(3-(phenylsulfonyl)propanamido) butyl)amino)octanoate (Compound 212). UPLC-MS: m / z (ESI) for C55H100N2O7S [M+H]+ expected: 934.47; observed: 934.32; 'H NMR (400 MHz, CDCh) 87.91 (dd, J= 7.3, 1.8 Hz, 2H), 7.66 (t, J= 7.5 Hz, 1H), 7.57 (t, J= 7.6 Hz, 2H), 6.77 (t, J= 5.4 Hz, 1H), 4.88 (p, J= 6.2 - 164 - LEGAL\113833648\6Hz, 1H), 4.07 (t, J= 7.1 Hz, 2H), 3.45 (dd, J= 9.1, 6.5 Hz, 2H), 3.17 (q, J = 6.1 Hz, 2H), 2.61 (dd, J= 9.0, 6.6 Hz, 2H), 2.43 - 2.34 (m, 6H), 2.28 (td, J= 7.6, 1.7 Hz, 4H), 1.68 - 1.37 (m, 18H), 1.36 - 1.17 (m, 45H), 0.94 - 0.83 (m, 12H).CggHtooN^OgSMW: 977.48Comoound 213

[0474] ethyl 3-((8-((8-((3-hexylnonyl)oxy)-8-oxooctyl)(4-(3-(phenylsulfonyl)propanamido) butyl)amino)octanoyl)oxy)decanoate (Compound 213). UPLC-MS: m / z (ESI) for C56H100N2O9S [M+H]+ expected: 978.48; observed: 978.3; 'H NMR (400 MHz, CDCh) 87.94 - 7.88 (m, 2H), 7.71 - 7.62 (m, 1H), 7.57 (dd, J= 8.5, 7.0 Hz, 2H), 6.76 (s, 1H), 5.28 - 5.17 (m, 1H), 4.09 (dq, J = 23.0, 8.0 Hz, 4H), 3.51 - 3.41 (m, 2H), 3.18 (q, J = 6.1 Hz, 2H), 2.67- 2.57 (m, 2H), 2.54 (t, J = 6.6 Hz, 2H), 2.38 (t, J = 7.7 Hz, 6H), 2.28 (td, J = 7.5, 5.0 Hz, 4H), 1.93 (dt, J = 12.7, 4.1 Hz, 2H), 1.76 - 1.46 (m, 8H), 1.45 - 1.37 (m, 2H), 1.35 - 1.03 (m, 50H), 0.88 (td, J= 6.8, 2.9 Hz, 9H).O.%Nil?C55H<,9N3O8SMW: 962.47Comnound 214

[0475] 1 -amino- 1 -oxoundecan-4-yl 8-((8-((3-hexylnonyl)oxy)-8-oxooctyl)(4-(3- (phenylsulfonyl)propanamido)butyl)amino)octanoate (Compound 214). UPLC-MS: m / z (ESI) for C55H99N3O8S [M+H]+ expected: 963.47; observed: 963.47; 'H NMR (400 MHz, CDCh) 8 7.94 - 7.87 (m, 2H), 7.71 - 7.62 (m, 1H), 7.62 - 7.53 (m, 2H), 6.92 (t, J= 5.4 Hz, 1H), 5.86 (s, 1H), 5.36 (s, 1H), 4.97-4.85 (m, 1H), 4.07 (t, J= 7.1 Hz, 2H), 3.51 - 3.41 (m, 2H), 3.18 (q, J = 6.2 Hz, 2H), 2.66 - 2.58 (m, 2H), 2.42 - 2.14 (m, 12H), 1.98 - 1.78 (m, 2H), 1.73-1.39 (m, 16H), 1.36 - 1.18 (m, 43H), 0.87 (td, J= 6.8, 2.9 Hz, 9H).

[0476] Following the same procedure as for Compound 210 the following compounds were prepared from 8-((8-((3-hexylnonyl)oxy)-8-oxooctyl)(4-(3-phenoxypropanamido)butyl) amino)octanoic acid (Compound XXXIII) and the corresponding alcohol starting materials.- 165 - LEGAL\113833648\6C8;3HwNgQ?SMW: W1.5SCompound 146

[0477] (Z)-3-ethyltetradec-8-en-5-yl 8-((8-((3-hexylnonyl)oxy)-8-oxooctyl)(4-(3-(phenylsulfonyl)propanamido)butyl)amino)octanoate (Compound 146). UPLC-MS: m / z (ESI) for C60H108N2O7S: [M+H]+expected: 1002.59; observed: 1002.30 'H NMR (400 MHz, CDCh) 8 7.97 (d, J = 7.7 Hz, 2H), 7.66 (t, J = 7.5 Hz, 1H), 7.63 - 7.51 (m, 2H), 5.45 - 5.21 (m, 4H), 4.07 (t, J = 7.1 Hz, 2H), 3.50 (t, J = 7.0 Hz, 2H), 3.31 (s, 2H), 3.09 (d, J = 7.1 Hz, 1H), 2.97 (s, 5H), 2.67 (q, J = 8.0 Hz, 3H), 2.49 (t, J = 7.4 Hz, 1H), 2.36 (q, J = 7.5 Hz, 1H), 2.27 (t, J = 7.5 Hz, 3H), 2.22 -2.13 (m, 1H), 2.09 - 1.97 (m, 2H), 1.92 (s, 1H), 1.57 (m, 12H), 1.29 (d, J = 30.3 Hz, 50H), 0.96 - 0.76 (m, 15H).CggH / iggNjOySMW: 989.58Comuound 147

[0478] 3-ethyltridecan-5-yl 8-((8-((3-hexylnonyl)oxy)-8-oxooctyl)(4-(3 -(phenylsulfonyl) propanamido)butyl)amino)octanoate. (Compound 147). UPLC-MS: m / z (ESI) for C59H108N2O7S:[M+H]+expected: 990.58; observed: 990.35. 'H NMR (400 MHz, CDCh) 88.10 - 7.87 (m, 2H), 7.80 - 7.61 (m, 1H), 7.58 (dd, J= 8.4, 6.8 Hz, 2H), 4.95 (t, J = 6.9 Hz, 1H), 4.07 (t, J = 7.1 Hz, 2H), 3.50 (q, J = 7.5 Hz, 2H), 3.31 (d, J = 5.9 Hz, 2H), 2.97 (s, 6H), 2.68 (t, J = 7.2 Hz, 2H), 2.26 (td, J = 7.5, 4.8 Hz, 2H), 2.02 - 1.86 (m, 3H), 1.70 (dt, J = 13.5, 3.9 Hz, 1H), 1.8 - 1.45 (m, 16H), 1.46 - 1.03 (m, 56H), 0.96 - 0.71 (m, 15H).C54H93N3O6SMW: 912.41Comnound 148

[0479] 3-hexylnonyl 8-((8-(((lR,3S,5r,7r)-adamantan-2-yl)amino)-8-oxooctyl)(4-(3-(phenyl sulfonyl)propanamido)butyl)amino)octanoate. (Compound 148). UPLC-MS: m / z (ESI) for C54H93N3O6S: [M+H]+expected: 913.41; observed: 913.25. 'H NMR (400 MHz, CDCh) 88.03 - - 166 - LEGAL\113833648\67.82 (m, 2H), 7.73 - 7.61 (m, 1H), 7.61 - 7.51 (m, 2H), 7.06 (s, 1H), 5.82 (d, J= 8.1 Hz, 1H), 4.06 (q, J = 8.4 Hz, 3H), 3.51 - 3.33 (m, 2H), 3.18 (q, J = 6.1 Hz, 2H), 2.74 -2.58 (m, 2H), 2.41 (s, 6H), 2.28 (t, J = 7.5 Hz, 2H), 2.19 (t, J = 7.5 Hz, 2H), 2.00 - 1.68 (m, 14H), 1.67 - 1.35 (m, 20H), 1.35 (m, 37H), 0.98 - 0.79 (m, 6H).C55H95N3O6SMW: 926.44Compound 149

[0480] 3-hexylnonyl 8-((8-((((3r,5r,7r)-adamantan-l-yl)methyl)amino)-8-oxooctyl)(4-(3-(phenylsulfonyl)propanamido)butyl)amino)octanoate (Compound 149)UPLC-MS: m / z (ESI) for C55H95N3O6S: [M+H]+expected: 927.44; observed: 927.13. 'H NMR (400 MHz, CDCh) 88.00 - 7.80 (m, 2H), 7.74 - 7.60 (m, 1H), 7.62 - 7.52 (m, 2H), 7.05 (s, 1H), 5.55 (s, 1H), 4.08 (t, J = 7.1 Hz, 2H), 3.59 - 3.35 (m, 2H), 3.18 (t, J = 6.0 Hz, 2H), 2.94 (d, J = 6.3 Hz, 2H), 2.68 - 2.51 (m, 2H), 2.37 (t, J = 5.3 Hz, 6H), 2.29 (t, J = 7.5 Hz, 2H), 2.20 (t, J = 7.6 Hz, 2H), 1.96 (d, J = 5.0 Hz, 3H), 1.78 - 1.37 (m, 32H), 1.37 - 1.10 (m, 27H), 0.99 - 0.69 (m, 6H)C50HS6N2O7SMW: 941.45Compound 150

[0481] 2-((3r,5r,7r)-adamantan-l-yl)ethyl 8-((8-((3-hexylnonyl)oxy)-8-oxooctyl)(4-(3-(phenylsulfonyl)propanamido)butyl)amino)octanoate (Compound 150)UPLC-MS: m / z (ESI) for C56H96N2O7S: [M+H]+expected: 942.45; observed: 942.25. ' H NMR (400 MHz, CDCh) 88.01 - 7.83 (m, 2H), 7.75 - 7.62 (m, 1H), 7.61 - 7.50 (m, 2H), 6.76 (s, 1H), 4.10 (dt, J = 16.2, 7.2 Hz, 4H), 3.55 - 3.40 (m, 2H), 3.18 (q, J = 6.1 Hz, 2H), 2.69 - 2.57 (m, 2H), 2.39 (s, 6H), 2.28 (td, J = 7.5, 3.0 Hz, 4H), 2.03 - 1.85 (m, 3H), 1.70 (d, J = 12.3 Hz, 4H), 1.67 -1.37 (m, 25H), 1.38 - 1.21 (m, 32H), 0.94 - 0.80 (m, 6H).- 167 - LEGAL\113833648\6C52H90N2O7SMW: 887.36Compound 157

[0482] bicyclo[2...

Claims

1. What is claimed is:

1. A compound having the formula of:Ri R?k2R2Ra i L-] o~ X-| L-| -]— RR-]N - LL — Nxr\|_| ' • r\ D l_20— X2— 1_21_ RR2(GI),ORR1R1 ^2R2L1Q-X-] L1— R RM;1M;2B1RH^T ^i\r ^LL _ i NZI-20- ^2 — h21- RR2(GII), wherein:RH is arylalkyl, aryl, heteroaryl, or heterocyclyl;each of Ri, Ri', R2, and R2', in each occurrence, is independently H, halo, C1-C3 alkyl, C3-C7 cycloalkyl, or -N(Ra)C(0)Rc; or Ri and Ri', or R2 and R2', together with their adjacent carbon atom, form a ring;nl and n2 are each independently 0, 1, 2, 3, 4, or 5;°A is absent, -O-, -S-, -S-S-, -C(Rb)(Rb')-, -C(O)-, -C(O)O-, -OC(O)-, -S(O)->0 0 o oV V '~'N— V >□— I- I s 1 3, -P(0)(0Ra)-, -N(Ra)-, -N(Ra)C(0)-, or -C(0)N(Ra)-;0 0yB is -C(O)-, -C(O)O-, -OC(O)-, -S(O)-> or;each of Ra and Ra', in each occurrence, is independently H, C1-C3 alkyl, or C3-C7 cycloalkyl;each of Rb and Rb', in each occurrence, is independently H, OH, C1-C3 alkyl, or C3-C7 cycloalkyl;Rc in each occurrence is independently H, alkyl, or -N(Ra)(Ra');Rq is H, alkyl, or aryl; optionally substituted with one or more of alkyl, halo, or -ORa; LL is C4 alkylene, C4 alkenylene, C4 alkynylene, or-(C(R3)(R3'))mi-Q-(C(R3)(R3'))m2-; or LL, together with Ra and the N atom connected to both B and Ra, forms a linker of the formulaeach of R3 and R3', in each occurrence, is independently H, C1-C3 alkyl, or C2-C3 alkenyl; Q is a divalent C3-C7 cycloalkyl, a divalent bridged cycle, or a divalent heterocycle; ml and m2 are each independently 0, 1, or 2;m3 is 0 or 1;m4 is 1 to 3;Xi and X2are each independently -C(O)-, -C(O)O-, -OC(O)-, -C(O)S-, -SC(O)-,- 244 - LEGAL\113833648\6-N(Ra)C(O)- or -C(O)N(Ra)-;Lio and L20 are each independently a C1-C12 alkylene, optionally substituted by C1-C3 alkyl;Ln and L21 are each independently absent or a Ci-Ce alkylene;t / Rm5E— \RR1 is -E-R30 or *Rl1;S / R2°| — E —RR2 is -E-R40 orRz1;E is absent, -CH(Ra)-, -CH(Ci-C4alkylene-CO(NH2))-, O, S, arylene, or heteroarylene; R30 and R40 each are independently H, branched or unbranched alkyl, branched or unbranched alkenyl, branched or unbranched alkynyl, bridged cyclyl, aryl, aralkyl, arylalkenyl, arylalkynyl, aralkoxy, heteroaryl, heteroarylalkyl, heteroarylalkenyl, or heteroarylalkynyl; optionally substituted with one or more of alkyl, halo, or -ORa;Rio and R20 each are independently H or a branched or unbranched, saturated or unsaturated Ci -C12 monovalent hydrocarbon chain; andR11 and R21 each are independently a branched or unbranched, saturated or unsaturated Ci -C12 monovalent hydrocarbon chain.

2. The compound of claim 1, whereinE is absent, -CH(CI-C3 alkyl), -CH(Ci-C4alkylene-CO(NH2))-, O, S, or phenylene; and R30 and R40 each are independently H, branched or unbranched alkyl, branched or unbranched alkenyl, branched or unbranched alkynyl, bridged cyclyl, phenyl, phenylalkyl, phenylalkynyl, phenyloxy, heteroaryl; optionally substituted with one or more of C1-C3 alkyl.

3. The compound of claim 1 or 2, wherein the compound has the formula ofA compound having formula of:R1 R / R2R2' Ra i x I.>1 1 O-X11°-R111 Mm AXn2 Jj_LL-N / L120-X120-R121RH^ \ ^L2I0— X2i0— R2nL20-A20_ L21 - << L220—^220“ R221 (PE-I) or - 245 - LEGAL\113833648\6Ri R?R2R2' L X L ^ / LI I°-XHO-RIII / Vn1 AX n2B__LL_N / L120-X120-R121RHX' ' '" A \ y i ^L21Q— X210— R211 / L20-A20— L21 Ra'L220-X220“R221Ty.(PE-II), wherein:RH is alkyl, arylalkyl, aryl, heteroaryl, or heterocyclyl;each of Ri, Ri', R2, and R2', in each occurrence, is independently H, halo, C1-C3 alkyl, C3-C7 cycloalkyl, or -N(Ra)C(O)Rc; or Ri and Ri', or R2 and R2', together with their adjacent carbon atom, form a ring;nl and n2 are each independently 0, 1, 2, 3, 4, or 5;0 0cA is absent, -O-, -S-, -S-S-, -C(Rb)(Rb')-, -C(O)-, -C(O)O-, -OC(O)-, -S(O)-,o o o oV■A, -P(O)(ORa)-, -N(Ra)-, -N(Ra)C(O)-, or -C(O)N(Ra)-;0 0yB is -C(O)-, -C(O)O-, -OC(O)-, -S(O)-, or;each of Ra and Ra', in each occurrence, is independently H, C1-C3 alkyl, or C3-C7 cycloalkyl;each of Rb and Rb', in each occurrence, is independently H, OH, C1-C3 alkyl, or C3-C7 cycloalkyl;Rc in each occurrence is independently H, alkyl, or -N(Ra)(Ra');Rq is H, alkyl, or aryl; optionally substituted with one or more of alkyl, halo, or -ORa; LL is C4 alkylene, C4 alkenylene, C4 alkynylene, or-(C(R3)(R3'))mi-Q-(C(R3)(R3'))m2-; or LL, together with Ra and the N atom connected to both B and Ra, forms a linker of the formulaeach of R3 and R3', in each occurrence, is independently H, C1-C3 alkyl, or C2-C3 alkenyl; Q is a divalent C3-C7 cycloalkyl, a divalent bridged cycle, or a divalent heterocycle; ml and m2 are each independently 0, 1, or 2;m3 is 0 or 1;m4 is 1 to 3;X10 and X20 are each independently -C(O)O-, or -OC(O)-;Xno, X120, X210, and X220 are each independently absent, -C(O)O-, or -OC(O)-, provided that at least one of Xno, Xi2o, X2io, and X220 is not absent;Lio and L20 are each independently a C1-C12 alkylene, optionally substituted by C1-C3 alkyl;- 246 - LEGAL\113833648\6Ln, L21, Luo, L120, L210, and L220 are each independently absent or a Ci-Ce alkylene; and Rm, R121, R211, and R221 each are independently H or a branched or unbranched, saturated or unsaturated Ci -C12 monovalent hydrocarbon chain;with the proviso that, when RH is alkyl, A is not absent and not CH2.

5. The compound of any one of claims 1-4, wherein the total number of atoms forming theL1o —— \chain connecting RH- toL20—is nine.

6. The compound of any one of claims 1-5, wherein LL is a C4 alkylene, C4 alkenylene, C4 alkynylene, or -(CH2)mi-Q-CH2)m2-,wherein:Q is a divalent C4-C6 cycloalkyl or a divalent bridged cycle; andml and m2 are each independently 0 or 1.

7. The compound of claim 6, wherein LL isThe compound of claim 3, wherein the compound has the formula of:wherein:Q’ is -CH2-CH2-, -CH=CH-, -C=C-, a divalent C3-C7 cycloalkyl or a divalent bridged cycle;- 247 - LEGAL\113833648\6ml and m2 are each independently 0 or 1;m3 is 0 or 1; and / orm4 is 1 to 3.The compound of claim 8, having the formula of:(GII-LIIa).

10. The compound of claim 1, wherein the compound has the formula of:^Liio-X1 1o-Riii RI <R2R2' Ra L10-X10-L1 1—L120~A120_ K121 >1YI ^*-210-^210-^211L20-A20-L21 - *-220— X220-R221 (PE-LI-1) ' R -L110-X110-R111L1O_XIO—L11 —L Av D120— 120-K121 RH-.L2O— Av2O. — L2^L210—X210—R211 I - L220-X220-R221 (PE-LI-2), or Ri R;R2R2^ / L110-X110-R11 1*-10—^10— *-11 - C. I V DL120~A120~ •'121, Q', N'..v| ^*-210-^210-^211L20“A20“L21 — Ra'L220-X220-R221 (PE-T IT) wherein:Q’ is -CH2-CH2-, -CH=CH-, -C=C-, a divalent C3-C7 cycloalkyl or a divalent bridged cycle;ml and m2 are each independently 0 or 1;m3 is 0 or 1; and / orm4 is 1 to 3.

11. The compound of claim 10, having the formula:..v| ^*-210-^210-^211L20- A20“L21 —*-220~ X220-R22I(PE-LIa), or- 248 - LEGAL\113833648\6(PE-LIIa).

12. The compound of any one of claims 1-11, wherein:each of Ri, Ri', R2, and R2', in each occurrence, is independently H, halo, CH3, -NHC(O) CH3; or R2 and R2', together with their adjacent carbon atom, form a ring;nl and n2 are each independently 0, 1, 2, or 3;A is absent, -O-, -S-, -S-S-, -CH2-, -C(O)-, -C(O)O-, -OC(O)-, -S(O)-,, -P(O)(ORa)-, -NH-, -N(CH3)-, -NHC(O)-, -CH(OH)-, or -C(O)N(Ra)-; wherein Ra is H or C1-C3 alkyl, and Rqis C1-C3 alkyl or phenyl optionally substituted with one or more alkyl or halo; and / orO OBis -C(O)-, -OC(O)-, or13. The compound of claim 12, wherein:each of Ri, Ri', R2, and R2' is H;nl is 0 or 1; andn2 is 0, 1, 2, or 3.

14. The compound of any one of claims 1-13, wherein B is C(O).O O_s15. The compound of any one of claims 1-13, wherein B is.

16. The compound of any one of claims 1-15, wherein:RH is arylalkyl, phenyl, biphenyl, pyrimidinyl or dioxohexahydropyrimidinyl, pyridyl, pyridinyl, pyrrolyl, imidazolyl, pyrazolyl, triazolyl, tetrazolyl, triazolyl, thiophenyl, indolyl, dioxoisoindolinyl, dihydrobenzodioxinyl, dihydrobenzodioxepanyl, dihydrobenzofuranyl, thienopyrrolyl, benzimidazolyl, dihydroindenyl, naphthalenyl, or oxathianyl or sulfoxidized oxathianyl;- 249 - LEGAL\113833648\6optionally substituted with one or more of alkyl, halo, haloalkyl, phenyl, heteroaryl, -ORa, -C(Rb)(Rb')ORa, -C(O)N(Ra)(Ra'), -SChRc, -NO2, -B(ORa)2, or -ON, wherein two or more substituents, when present, may form a fused ring with the main ring of RH;wherein:Rc is H, alkyl, or -N(Ra)(Ra'), andRd is H, alkyl, or phenyl.(R6)t(R6)tR517. The compound of claim 16, wherein RH is(R6)t(R6)t(R6)t(R6)t, or; wherein:R4 is H or C1-C3 alkyl;each of R5and Rs', in each occurrence, is independently aryl or heteroaryl, optionally substituted with one or more of alkyl, halo, haloalkyl, or -ORe, andRe is, in each occurrence, independently H, alkyl, halo, haloalkyl, phenyl, heteroaryl, -OR7, -(C(R4)(R4'))SOR7, -C(O)N(R4)(R4')> -NO2, -ON, -SO2R4, -B(OR7)2, or =0;R7is H, C1-C3 alkyl, or phenyl;I FR)' —zis a 5-, 6-, or 7- member aromatic or non-aromatic ring fused to the phenyl ring, optionally containing 1 to 2 of O, S, or N on the ring;( HR )is a 5- or 6- member heteroaromatic or heterocycle containing 1 to 4 of S or N on the ring; ands and t are each independently 1 to 3.(R6)t18. The compound of claim 16, wherein RH is; wherein:Re IS H, alkyl, halo, haloalkyl, -OH, -0CH3, -C(0)NH2, -NO2, -ON, -SO2CH3 or -B(0H)2.- 250 - LEGAL\113833648\6LEGAL\113833648\6OHHFri?;H '- 252 - LEGAL\113833648\6H- 253 - LEGAL\113833648\6wherein I represents the connection to LL.

21. The compound of any one of claims 1-3, 5-9, and 12-20, wherein:Xi and X2 are each independently -C(O)O-, -OC(O)-, -NHC(O)-, or -C(O)NH-;Lio and L20 are each independently a C4-C8 alkylene, optionally substituted by methyl; Ln and L21 are each independently absent or a C1-C3 alkylene;- 254 - LEGAL\113833648\6Rio and R20 are each independently H or a branched or unbranched, saturated or unsaturated C3 -C10 monovalent hydrocarbon chain; and / orR11 and R21 are each independently a branched or unbranched, saturated or unsaturated C3 -C10 monovalent hydrocarbon chain.

22. The compound of claim 21, wherein:Lio and L20 are each independently a linear C5-C7 alkylene;Ln and L21 are each independently absent or a C2 alkylene;Rio and Rn are each independently a linear C4-C8 alkyl; or Rio is H, and Rn is a linear C6-C9 alkyl or C6-C9 alkenyl; and / orR20 and R21 are each independently a linear C4-C8 alkyl; or R20 is H, and R21 is a linear C6-C9 alkyl or C6-C9 alkenyl.

23. The compound of claim 22, wherein:Lio and L20 are each independently a linear C5 or C7 alkylene;Ln and L21 are each independently absent or a C2 alkylene;Rio and Rn are each independently a linear C4, Ce, or Cs alkyl; or Rio is H, and Rn is a linear Cs alkyl or Cs alkenyl; and / orR20 and R21 are each independently a linear C4, Ce, or Cs alkyl; or R20 is H, and R21 is a linear Cs alkyl or Cs alkenyl.

24. The compound of any one of claims 21-23, wherein one or more of the following conditions are met:Xi and X2 are different;Ln and L21 are different;Lio and L20 are different;Rio and Rn are different;R20 and R21 are different; andc ^xR10 _ ^R201is different than^21.

25. The compound of any one of claims 21-23, wherein one of Ln and L21 is absent, and the other one of Ln and L21 is a Ci-Ce alkylene.—; — 1-11 — RR1 26. The compound of any one of claims 21-25, wherein each of the tail groups!— L-21 — RR2and l is independently:- 255 - LEGAL\113833648\6the connection to Xi or X2.

27. The compound of any one of claims 1-2, 5-7, and 12-20, wherein the tail- 256 - LEGAL\113833648\6LEGAL\113833648\6LEGAL\113833648\6, wherein represents the connection to LL.

29. The compound of any one of claims 1-3, 5-9, and 12-20, wherein the compound has the formula of:o o(GII-TI);orwherein:ulO and u20 are each independently 4 to 8;ul 1 and u21 are each independently 0 to 3;Rio and Rn are each independently a linear C4-C8 alkyl; or Rio is H or a linear C1-C4 alkyl, and Rn is a linear C6-C9 alkyl or C6-C9 alkenyl; and / orR20 and R21 are each independently a linear C4-C8 alkyl; or R20 is H or a linear C1-C4- 259 - LEGAL\113833648\6alkyl, and R21 is a linear C6-C9 alkyl or C6-C9 alkenyl;optionally wherein one of ul 1 and u21 is 0, and other one of ul 1 and u21 is 1 to 3.

30. The compound of claim 29, whereinRio and Ru are the same; and / orR20 is H or C1-C4 alkyl, and R21 is a linear C6-C9 alkyl.

31. The compound of claim 29 or 30, having the formula of:

33. The compound of claim 1, wherein the compound is a compound in Table 1, Table 2, Table 3, or Table 4.

34. The compound of any one of claims 1-3, 5-9, and 21-32, wherein the compound has the OR2 R2Ru — A'^20formula of '21(GI-SI), wherein:- 260 - LEGAL\113833648\6RH is aryl or heteroaryl, optionally substituted with one or more of alkyl, hydroxyalkyl, alkoxyalkyl, halo, haloalkyl, -NO2, -C≡N, -ORa, -C(0)NH2, -SChRd, or -B(ORa)2, or aryl optionally fused with a non-aromatic ring;n2 is 0, 1, or 2;Ra and Rd are each independently H or C1-C3 alkyl;O O. VA is -CH2-, -CH(OH)-, -C(O)-, -O-, -S-,, -NH-, or -NHC(O)-;o oBis -C(O)-, or;each of R2 and R2', in each occurrence, is independently H, CH3, -NHC(O) CH3; or R2 and R2', together with their adjacent carbon atom, form a C3 ring;ulO and u20 are each independently 4 to 8;ul 1 and u21 are each independently 0 to 3;Rio and Rn are each independently a linear C4-C8 alkyl; or Rio is H or a linear C1-C4 alkyl, and Rn is a linear C6-C9 alkyl or C6-C9 alkenyl; and / orR20 is H or C1-C4 alkyl, and R21 is a linear C6-C9 alkyl;optionally wherein one of ul 1 and u21 is 0, and other one of ul 1 and u21 is 1 to 3.

35. The compound of claim 34, wherein:RH is phenyl optionally substituted with one or more of C1-C3 alkyl, aryl, hydroxymethyl, halo, haloalkyl, -NO2, -C≡N, -OCH3, -C(O)NH2, -SO2CH3, or -B(OH)2; dihydrobenzodioxinyl; or tetrazolyl, thiophenyl, pyridinyl, pyrrolyl, pyrazolyl, pyrazinyl, or pyrimidinyl, each optionally substituted with one or more methyl;Ra is H;Rd is H or methyl;nl is 0; and n2 is 0 or 2.- 261 - LEGAL\113833648\600 H0 0 H37. The compound of claim 1 or 34, wherein the compound is Lipid No. 4, 18, 26, 38, 40, 45, 47, 48, 53, 55, 57, 60, 62, 64, 65, 84, 88, 95, 97, 101, 106, 111, 114, 115, 116, 125, 130, 133, 181, 242, or 243.

38. The compound of claim 1 or 34, wherein the compound is Lipid No. 1, 4, 5, 8, 10, 13, 14, 17, 18, 20, 24, 25, 26, 27, 30, 31, 32, 38, 40, 45, 46, 47, 50, 57, 60, 62, 88, 104, 106, 107, 123, 127, 128, 134, 242, or 243.- 262 - LEGAL\113833648\639. The compound of claim 1 or 34, wherein the compound is lipid No. 1, 8, 13, 17, 18, 26, 27, 32, 40, 45, 46, 50, 60, 88, 104, 106, 107, 123, 127, or 134.

40. The compound of claim 39, wherein the compound is 40, 26, 18, 45, 60, or 38.

41. The compound of any one of claims 4-7 and 10-20, wherein:Lio and L20 are each independently a C4-C8 alkylene, optionally substituted by methyl; Ln, L21, Luo, L120, L210, and L220 are each independently absent or a C1-C3 alkylene; and / orRm, R121, R211, and R221 are each independently H or branched or unbranched, saturated or unsaturated Ci -C10 monovalent hydrocarbon chain.

42. The compound of claim 41, wherein:Lio and L20 are each independently a linear C5-C7 alkylene; and / orLn, L21, Luo, L120, L210, and L220 are each independently absent or a C1-C2 alkylene.

43. The compound of claim 41 or 42, wherein:X10 and X20 are -C(O)O-; and / or(i) one of Xno, Xi2o, X2io, and X220 is -C(O)O- or -OC(O)-, and the other three are absent; (ii) two of Xno, X120, X210, and X220 are each independently -C(O)O- or -OC(O)-, and the other two are absent;(iii) three of Xno, Xi2o, X2io, and X220 are each independently -C(O)O- or -OC(O)-, and the other one is absent; or(iv) all four of Xno, Xi2o, X2io, and X220 are each independently -C(O)O- or -OC(O)-.

44. The compound of claim 43, wherein:X10 and X20 are -C(O)O-; and(i) Xno and X120 are absent, and one of X210 and X220 is -C(O)O- or -OC(O)-, and the other is absent; or(ii) Xno and X120 are absent, and X210 and X220 are each independently -C(O)O- or -OC(O)-; or(iii) one of Xno and X120 is absent, and the other is -C(O)O- or -OC(O)-; and X210 and X220 are each independently -C(O)O- or -OC(O)-.

45. The compound of any one of claims 41-44, wherein:X10 and X20 are -C(O)O-; and each of Xno, Xi2o, X2io, and X220, if present, is -OC(O)-.

46. The compound of any one of claims 41-44, wherein:X10 and X20 are -C(O)O-; and each of Xno, Xi2o, X2io, and X220, if present, is -C(O)O-.- 263 - LEGAL\113833648\647. The compound of any one of claims 41-46, wherein in each of the tail groups,- 264 - LEGAL\113833648\6connection to LL.

49. The compound of any one of claims 4-5, 10-11, and 41-46, wherein the compound has theformula of wherein:RH is alkyl, aryl, or heteroaryl, optionally substituted with one or more of alkyl, hydroxyalkyl, alkoxyalkyl, halo, haloalkyl, -NO2, -C≡N, -ORa, -C(O)NH2, -SChRd, or -B(ORa)2, or aryl optionally fused with a non-aromatic ring;n2 is 0, 1, or 2,Ra and Rd are each independently H or C1-C3 alkyl;O OAis -CH2-, -CH(OH)-, -C(O)-, -O-, -S-,, -NH-, or -NHC(O)-;each of R2 and R2', in each occurrence, is independently H, CH3, -NHC(O)CH3; or R2 and R2', together with their adjacent carbon atom, form a C3 ring.

50. The compound of claim 49, wherein:RH is methyl or phenyl optionally substituted with one or more of C1-C3 alkyl, aryl, hydroxymethyl, halo, haloalkyl, -NO2, -C≡N, -OCH3, -C(O)NH2, -SO2CH3, or -B(OH)2; dihydrobenzodioxinyl; or tetrazolyl, thiophenyl, pyridinyl, pyrrolyl, pyrazolyl, pyrazinyl, or pyrimidinyl, each optionally substituted with one or more methyl groups;Ra is H;Rd is H or methyl;nl is 0; andn2 is 0 or 2.- 265 - LEGAL\113833648\651. The compound of claim 4 or 49, wherein the head group owherein I represents the connection to LL.

52. The compound of claim 4 or 49, wherein the compound is a compound in Table 5.

53. The compound of claim 4 or 49, wherein the compound is Lipid No. 159, 224, 240, 237, 225, 238, 239, 208, 215, 217, or 174.

54. A lipid composition comprising the compound according to any one of claims 1-53, and a lipid component; optionally the lipid composition is a lipid nanoparticle.

55. The lipid composition of claim 54, wherein the lipid component comprises a sterol, a PEG-modified lipid, and / or a helper lipid.

56. The lipid composition of claim 55, wherein the helper lipid is l,2-dioeoyl-sn-glycero-3-phosphoethanolamine (DOPE), l,2-dioleoyl-sn-glycero-3-phosphocholine (DOPC),2-distearoyl-sn-glycero-3-phosphocholine (DSPC), dimethyldioctadecylammonium (DDAB), di dodecyldimethylammonium bromide (DDAB), l,2-di-O-octadecenyl-3 -trimethylammonium propane (DOTMA), l,2-dioleoyl-3-trimethylammonium-propane (DOTAP), 18:1 PA, N-methyldioctadecylamine (MDOA), N, N-dicotadecylaniline, sn-(3-myristoyl-2-hydroxy)-glycerol-l-phospho-sn-3'-(l',2'-dimyristoyl)-glycerol (14:0 Hemi BMP), l,2-dimyristoyl-sn-glycero-3-phosphate (DMPA; 14:0 PA), l,2-dioleoyl-sn-glycero-3 -phosphate (DOPA; 18:1 PA), 1,2-distearoyl-sn-glycero-3 -phosphate (DSPA; 18:0 PA), or a combination thereof.

57. The lipid composition of claim 55, wherein the sterol is cholesterol, a cholesterol derivative, or a combination thereof.

58. The lipid composition of claim 55, wherein the PEG-modified lipid is PEG-DMG, PEG-DSG, PEG-PE, PEG-DSPE; optionally the PEG-modified lipid is l,2-dimyristoyl-sn-glycero-3-phosphoethanolamine-N-[methoxy(polyethyleneglycol)-2000(DMPE-PEG2K; 14:0 PEG2K PE), l,2-distearoyl-sn-glycero-3-phosphoethanolamine-N- [methoxy (polyethyl eneglycol)-2000 (DSPE-PEG2K; 18:0 PEG2KPE), 1,2-dimyristoyl-sn-glycero-3 -methoxypoly ethylene glycol- 266 - LEGAL\113833648\6(DMG-PEG 2K), 2- [(polyethylene glycol)-2000]-N, N-ditetradecylacetamide (ALC-0159), distearoyl-rac-glycerol-PEG2K (DSG-PEG 2K), or a combination thereof.

59. The lipid composition of any one of claims 55-58, wherein the lipid composition comprises:about 20-60 mol% of the compound,about 7-50 mol% of the helper lipid,about 5-70 mol% of the sterol, andabout 0.5-3 mol% of the PEG-modified lipid.

60. The lipid composition of any one of claims 54-59, having a diameter of about 20-200 nm, and / or a poly dispersity index of about 0.3 or less.

61. The lipid composition of any one of claims 54-60, further comprising a targeting moiety, conjugated to one or more lipid components of the lipid composition.

62. The lipid composition of claim 61, wherein the targeting moiety is a peptide, polypeptide, protein; a carbohydrate or polysaccharide; or a small molecule targeting ligand, optionally wherein the targeting moiety is covalently attached to the one or more lipid components of the lipid composition.

63. The lipid composition of claim 61 or 62, wherein the conjugation of the targeting moiety to the lipid composition is carried out by functionalizing the targeting moiety with a first functional group, and functionalizing a lipid component of the lipid composition with a second functional group, wherein the first and second functional groups are reactive or interactive to each other to form the conjugation.

64. The lipid composition of claim 61 or 62, wherein the lipid composition contains a PEG-modified lipid, and wherein functionalizing the lipid component is carried out by functionalizing the PEG-modified lipid.

65. The lipid composition of claim 64, wherein the lipid composition contains two or more PEG-modified lipids, and wherein at least one PEG-modified lipid is not functionalized and at least one PEG-modified lipid is functionalized.

66. The lipid composition of claim 65, wherein the molar ratio of the PEG-modified lipid that is not functionalized to the functionalized PEG-modified lipid is 1: 1 or greater, such as about 2: 1- 267 - LEGAL\113833648\6or greater, about 5:1 or greater, about 10:1 or greater, about 15:1 or greater, about 20:1 or greater, about 30: 1 or greater, about 40: 1 or greater, about 50: 1 or greater, about 60: 1 or greater, about 70: 1 or greater, about 80: 1 or greater, about 90: 1 or greater, about 100: 1 or greater, about 150:1 or greater, about 200: 1 or greater, or about 300 or greater.

67. The lipid composition of any one of claims 54-66, further comprising a therapeutic agent; optionally wherein the therapeutic agent is a nucleic acid molecule, protein or peptide, carbohydrate or glycoprotein, small molecule, or any combination thereof.

68. The lipid composition of claim 67, wherein the therapeutic agent is DNA or RNA.

69. The lipid composition of claim 68, wherein the therapeutic agent is an mRNA.

70. A pharmaceutical composition comprising the lipid composition of any one of claims 67- 69, and a pharmaceutically acceptable excipient.

71. A method for delivering a therapeutic agent to a cell or a subject, comprising: contacting the cell, or administering to the subject, the lipid composition of any one of claims 54-69, or the pharmaceutical composition of claim 70.

72. The method of claim 71, wherein the therapeutic agent is delivered to kidney, liver, splenic, lymphatic, or marrow cells of the subject.

73. The method of claim 72, wherein the therapeutic agent is delivered to one or more of kidney endothelial cells, kidney epithelial cells, kidney dendritic cells, kidney B cells, kidney T cells, kidney natural killer (NK) cells, kidney macrophages, or other immune cells within the kidney, kidney collecting duct principal cells, kidney distal convoluted tubule cells, kidney glomerular endothelial cells, kidney intercalated cells, kidney interstitial fibroblasts,kidney mesangial cells, kidney parietal epithelial cells, kidney pericytes, kidney podocytes, kidney proximal tubule cells, kidney thick ascending limb cells, or kidney vascular endothelial cells; liver endothelial cells, hepatocytes, liver macrophages, liver dendritic cells, liver Kupffer cells, liver B cells, liver T cells, or other immune cells within the liver; spleen dendritic cells, spleen neutrophils, spleen macrophages, spleen B cells, spleen T cells, spleen natural killer (NK) cells, or other immune cells within the spleen; lymphatic dendritic cells, lymphatic neutrophils, lymphatic macrophages, lymphatic B cells, lymphatic T cells, or lymphatic natural killer (NK) cells of the subject.- 268 - LEGAL\113833648\674. The method of claim 72, wherein the therapeutic agent is delivered to stem-like cells (e.g., bone marrow stem-like cells), and / or hematopoietic stem cells (HSCs) (e.g., bone marrow HSCs).

75. A method for delivering of a therapeutic agent to a subject with specificity to kidney, comprising:administering to the subject the lipid composition of any one of claims 54-69, or the pharmaceutical composition of claim 70, wherein the compound is lipid No. 1, 8, 13, 17, 18, 26, 27, 32, 40, 45, 46, 50, 60, 88, 104, 106, 107, 123, 127, or 134;preferably, the compound is lipid No. 40, 26, 18, 45, 60, or 38.

76. The method of claim 75, wherein the therapeutic agent is delivered to one or more of kidney endothelial cells, kidney epithelial cells, kidney dendritic cells, kidney B cells, kidney T cells, kidney natural killer (NK) cells, kidney macrophages, or other immune cells within the kidney, kidney collecting duct principal cells, kidney distal convoluted tubule cells, kidney glomerular endothelial cells, kidney intercalated cells, kidney interstitial fibroblasts,kidney mesangial cells, kidney parietal epithelial cells, kidney pericytes, kidney podocytes kidney proximal tubule cells, kidney thick ascending limb cells, or kidney vascular endothelial cells.

77. A method of treating a disease or disorder in a subject in need, the method comprising: administering to the subject the lipid composition of any one of claims 54-63, or the pharmaceutical composition of claim 64, optionally wherein the disease or disorder is an infectious disease, a cancer, a proliferative disease, a genetic disease, an autoimmune disease, diabetes, a neurodegenerative disease, a cardiovascular disease, a reno-vascular disease, or a metabolic disease.

78. The method of claim 77, wherein the disease or disorder is cystic fibrosis, hemophilia A, hemophilia B, thalassemia, anemia or other blood disorders, AIDS, Alzheimer's disease, Parkinson's disease, Huntington's disease, amyotrophic lateral sclerosis, epilepsy, and other neurological disorders, cancer, diabetes mellitus, muscular dystrophies (e.g., Duchenne, Becker), Hurler's disease, adenosine deaminase deficiency, metabolic defects or disorders, retinal degenerative diseases or other diseases of the eye), mitochondriopathies (e.g., Leber's hereditary optic neuropathy (LHON), Leigh syndrome, and subacute sclerosing encephalopathy), myopathies (e.g., facioscapulohumeral myopathy (FSHD) and cardiomyopathies), diseases of solid organs (e.g., brain, liver, kidney, heart), metabolic diseases or disorders (e.g., Fabry disease, Gaucher disease, phenylketonuria (PKU), glycogen storage disease); urea cycle diseases or disorders (e.g., ornithine transcarbamoylase (OTC) deficiency); lysosomal storage diseases or disorders (e.g., metachromatic leukodystrophy (MLD), mucopolysaccharidosis Type II (MPSII;- 269 - LEGAL\113833648\6Hunter syndrome)); liver diseases or disorders (e.g., progressive familial intrahepatic cholestasis (PFIC); cancers, tumors, or genetic diseases or disorders (e.g., cystic fibrosis).

79. A method of treating a subject with a kidney disease or kidney disorder, the method comprising: administering to the subject the lipid composition of any one of claims 54-69, or the pharmaceutical composition of claim 70, wherein the compound is lipid No. 1, 8, 13, 17, 18, 26, 27, 32, 40, 45, 46, 50, 60, 88, 104, 106, 107, 123, 127, or 134;preferably, the compound is lipid No. 40, 26, 18, 45, 60, or 38.

80. The method of claim 79, wherein the kidney disease or kidney disorder is Alport syndrome, Bartter syndrome types 1-4, congenital nephrotic syndrome, cystinuria, cystinosis, Dent disease types 1 and 2, distal renal tubular acidosis, Fabry disease, familial amyloidosis, Gitelman syndrome, Liddle syndrome, Lowe syndrome, nephronophthisis (NPHP), autosomal dominant polycystic kidney disease, autosomal recessive polycystic kidney disease, tubulointerstitial kidney disease, proximal renal tubular acidosis, thin basement membrane disease, tubulointerstitial kidney disease, tuberous sclerosis complex (TSC), or APOL1 mediated kidney disease.- 270 - LEGAL\113833648\6

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