FURAN-DERIVED LIPID NANOPARTICLES FOR TRANSPORTING mRNA TO THE CENTRAL NERVOUS SYSTEM

Abstract

A new class of multi-functional lipid nanoparticles comprising an ionizable lipid including furan-derived lipid is disclosed. The disclosed nanoparticles bypass the blood-brain barrier via a lymphatic pathway, and travel from the deep cervical lymph nodes (dCLNs) to the meninges to deliver messenger RNA (mRNA) payload to the central nervous system (CNS). Compared to currently utilized nanoparticles for drug delivery, the disclosed nanoparticles leverage the meningeal lymphatic vessel (MLVs) route to achieve efficient mRNA delivery to the brain.

Description

Docket No. 084284.00356FURAN-DERIVED LIPID NANOPARTICLES FOR TRANSPORTING mRNA TO THE CENTRAL NERVOUS SYSTEMCROSS REFERENCE TO RELATED APPLICATION

[0001] The application claims the benefit of priority under 35 U. S. C. § 119(e) to U. S.Provisional Application No. 63 / 776,373, filed March 24, 2025, the entire disclosure of which is incorporated by reference herein.FIELD

[0002] The present disclosure relates in general to formulations of lipid nanoparticles. The disclosure relates in particular to intracellular delivery of messenger RNA to the central nervous system.BACKGROUND

[0003] Messenger RNA (mRNA) has been applied to many therapeutic fields such as vaccines, cancer immunotherapies, and protein replacement therapies. However, effective delivery of mRNA to the brain continues to be a major challenge. Previously, different routes of administration have been studied for drug delivery to the brain, such as intraparenchymal, intrathecal, intranasal, and systemic administrations. Intraparenchymal injection is currently employed in clinical trials because it directly delivers therapeutics to diseased sites. But this approach is highly invasive. Less invasive than intraparenchymal injection, intrathecal administration provides a direct route for transporting therapeutic agents to the cerebrospinal fluid (CSF) and then to the central nervous system (CNS).

[0004] However, repeated dosing via the intrathecal route often reduces patient compliance. The intranasal route presents a non-invasive alternative to bypass the blood-brainDocket No. 084284.00356barrier (BBB). For example, mRNA-loaded cationic liposomes have been delivered into some regions of the brain by intranasal administration, such as the cortex, striatum, and midbrain. However, this method is challenged by delivery efficiency due to nasal passage conditions and potential drug entrapment within the olfactory mucosa. Lastly, intravenous injection is currently the most clinically used administration route and is suitable for repeated dosing. However, the limited permeability of the BBB may result in suboptimal CNS delivery efficiency.

[0005] Given the foregoing, there is still an urgent need to develop innovative strategies to achieve potent delivery to the brain. This document describes compositions and methods that address some or all of the problems described above.SUMMARY

[0006] The present disclosure provides novel compounds and compositions involving the same.

[0007] A first aspect of the disclosure relates to compounds of Formula (I);, R2" / L\R1' A BXR2'(I),

[0008] wherein A is a furan or tetrahydrofuran; B is a bond, furan or tetrahydrofuran; L1and L2are independently ester (-O-(C=O)- or -(C=O)-O-), ether, amide (-(C=O)-N- or -N-(C=O)-), amino-Ci-5 alkylene, or oxygen-Ci-5 alkylene, wherein amino-Ci-5 alkylene and oxygen-Ci-5 alkylene may be optionally substituted with one or more selected from the group consisting of Ci-5 alkyl, amino, and hydroxide, and wherein amino-Ci-5 alkylene, or oxygen-Ci-5 alkylene are attached to A or B at alkylene end;Docket No. 084284.00356

[0009] R1, R1, R2and R2are independently hydrogen, a void or a linear or branched Ci-36 alkyl, optionally inserted with one or more ester, carbonate(-O-(C=O)-O-), amino, amide, or oxygen (-O-), and optionally substituted with one or more selected from the group consisting of halo-Ci-5 alkyl, C1-5 alkyl, amino, hydroxide, and halogen; V is a bond, a linear or branched C1-5 alkylene optionally substituted with one or more C1-5 alkyl, (-[C(L4)(L5)]-), C3-6 cycloalkyl, or Ci-5 alkylene-D, wherein D is E-L3(R3)(R3”); L4and L5are independently C1-5 alkyl or a hydrogen; E is a furan or tetrahydrofuran; L3is an ester, ether, amide, amino-Ci-5 alkylene, or oxygen-Ci-5 alkylene, wherein amino-Ci-5 alkylene and oxygen-Ci-5 alkylene may optionally be substituted with one or more selected from the group consisting of C1-5 alkyl, amino, and hydroxide, and wherein amino-Ci-5 alkylene, or oxygen-Ci-5 alkylene are attached to E at alkylene end; R3and R3are independently hydrogen, a void or a linear or branched Ci-36 alkyl, optionally inserted with one or more selected from the group consisting of ester, carbonate, amino, amide, and oxygen, and optionally substituted with one or more selected from the group consisting of halo-Ci-5 alkyl, C1-5 alkyl, amino, hydroxide, and halogen; wherein the void is represented by a dotted line, wherein when R1or R2is not void, it is connected via a single bond;

[0010] wherein when L1is amino-Ci-s alkylene or an amide, L1is bonded via nitrogen to both R1and R1”, wherein one or both R1or R1can be hydrogen, when L1is ester, ether, or an oxygen-Ci-5 alkylene, only R1is bonded via oxygen to L1and R1is void, when L2is amino-Ci-5 alkylene or amide, L2is bonded via nitrogen to both R2and R2, wherein one or both R2or R2can be hydrogen, when L2is ester, ether, or an oxygen-Ci-5 alkylene, only R2is bonded via oxygen to L2and R2is void, when L3is amino-Ci-5 alkylene or amide, L3 is bonded via nitrogen to both R3and R3, wherein one or both R3or R3can be hydrogen, and when L3is ester, ether, or an oxygen-Ci-5 alkylene, only R3is bonded via oxygen to L3and R3is void.Docket No. 084284.00356

[0011] In some embodiments, when B is a furan or a tetrahydrofuran, A is connected to L1at ring position 2 of A; A is connected to V at ring position 5 of A;

[0012] A, and

[0013] B is connected to L1at ring position 5 of B; and B is connected to V at ring position 2 ofB;12

[0014] wherein F1and F2are independently selected from the group consisting of a linear or branched Ci-is alkyl, Ci-i8 alkoxyl-alkylene, hydrogen, halo-Ci-5 alkyl, C1-5 alkyl, Ci-6 cycloalkyl-Ci-i8 alkylene, amino, hydroxide, and halogen.

[0015] In some embodiments, V is -[C(L4)(I?)]-; L4and I? are independently C1-5 alkyl or a hydrogen; L1and L2are independently amino-Ci-5 alkylene optionally substituted with one or more selected from the group consisting of alkyl, amino, and hydroxide; and R1, R1”, R2, and R2are independently hydrogen, a linear or branched Ci-36 alkyl, optionally inserted with one or more selected from the group consisting of ester, carbonate, amino, amide, and oxygen, and optionally substituted with one or more selected from the group consisting of halo-Ci-5 alkyl, C1-5 alkyl, amino, hydroxide, and halogen.Docket No. 084284.00356Qi CH ^ / \ QI z ^ / ai

[0016] In some embodiments, V is a bond; L1and L2are independently amino-Ci-5 y c> alkylene optionally substituted with one or more selected from the group consisting of alkyl, amino, and hydroxide; and R1, R1”, R2and R2are independently hydroge)nx o, a linear or branched Ci- 6 l y, p i n l y n ^z z"3 a k l o t o a l i serted with one or more selected from the gr7oup consisting of ester, / =\_ or orcarbonate, amino, amide, and oxygen, and optionally substituted with one or more selected from the group consisting of halo-Ci-5 alkyl, C1-5 alkyl, amino, hydroxide, and halogen, wherein A is connected to B at ring position 5 of A and ring position 2 of B, A is connected to L1at ring position 2 of A, and B is connected to L2at ring position 5 of B.1 1L1^2 / °X5 2 / OS5,L2 / / V 73W4 3W4F1F2F1F2 / A B

[0017]

[0018] wherein F1and F2are independently selected from the group consisting of a linear j cCor bran^c 0hed Ci-18 alkyl, Ci-18 alkoxyl-alkylene, hydrogen, halo-Ci-5 alkyl, C1-5 alkyl, C1-6 0cyclo / alkyl-Ci-18 alkylene, amino, hydroxide, and halogen.

[0019] In some embodiments, the compound of Formula (I) may have a formula:R1\ I Xu l / Docket No. 084284.00356M CM C M C CM Qi Qi CMM C QivQiM C CM / Q » / QixC / Di 7 z °1 o LA\2L °^ ^zzoQ oF £ (£7 / TJO iO |VM CV / Qi“■I z ZM CQiM C° ^ \ / QiO Cj A z- oiQo > oO oo ^x / -°JL - ° *rjoZ^^\Z / -r 7J *>z Nz^tr Qi R3\ZR3" Qi •^WNAz ' Rl'\RRI"\ oiR2R1"\ RM C2’ 2' 0i z CMM C. N J / / \ / Qxoi N / Qi QiM C CMRi \ / OVZ>^C\^ R:1yj X zX y# tXR / °- W-I-IO — ^1-101-101-10 (9^10no / X Xx°_x / z^ NPXR2"k!2.zi X o / -- a: ZE / - Qi ' * *1-10 ^*1-10tr \Qi QiR1"\ R2R1"\ R2R \ _ ^Ov / Nx,Ri y-< R2R1 V“< R2' *1-10 ' *1-10 ' '1-10wherein R1, R1, R2, R2, R3, or R3are independently selected from:OH / \^g'O / X'Ox^ o> AoArDocket No. 084284.00356, wherein f, g, and h are independently between 1-36 and the sum of g and h is between 2- 36, and one or more carbon atom on –(CH2)f–, –(CH2)g– or –(CH2)h– is optionally substituted with one or more selected from the group consisting of halo-Ci-5 alkyl, C1-5 alkyl, amino, hydroxide, and halogen.

[0020] In another aspect, another subset of compounds of Formula (I) includes the Formula (IA):RI;, R2"R1 R(IA),

[0021] wherein V and B are each a bond to form a single bond between A and L2, A is a furan or tetrahydrofuran; L1and L2are independently ester, ether, amide, amino-Ci-5 alkylene, or oxygen-Ci-5 alkylene, wherein amino-Ci-5 alkylene and oxygen-Ci-5 alkylene may be optionally substituted with one or more selected from the group consisting of C1-5 alkyl, amino, and hydroxide, and wherein amino-Ci-5 alkylene, or oxygen-Ci-5 alkylene are attached to A at alkylene end; R1, R1”, R2and R2are independently hydrogen, a void or a linear or branched Ci-36 alkyl, optionally inserted with one or more selected from the group consisting of ester, carbonate, amino, amide, and oxygen, and optionally substituted with one or more selected from the group consisting of halo-Ci-5 alkyl, C1-5 alkyl, amino, hydroxide, and halogen, wherein when L1is amino-Ci-5 alkylene or amide, L1is bonded via nitrogen to both R1and R1, wherein one or both of R1or R1can be hydrogen, when L1is ester, ether, or an oxygen-Ci-5 alkylene, only R1is bonded via oxygen to L1and R1is void, when L2is amino-Ci-5 alkylene or amide, L2is bonded via nitrogen to both R2and R2”, wherein one or both of R2or R2can be hydrogen, when L2is ester, ether, or anDocket No. 084284.00356oxygen-Ci-5 alkylene, only R2is bonded via oxygen to L2and R2is void, A is connected to L1at ring position 2 of A, and A is connected to L2at ring position 5 of A.12V7F1

[0022]

[0023] wherein F1and F2are independently selected from the group consisting of a linear or branched Ci-is alkyl, Ci-is alkoxyl-alkylene, hydrogen, halo-Ci-5 alkyl, C1-5 alkyl, Ci-6 cycloalkyl-Ci-i8 alkylene, amino, hydroxide, and halogen.

[0024] In some embodiments, L1is amino-Ci-5 alkylene optionally substituted with one or more selected from the group consisting of C1-5 alkyl, amino, and hydroxide; L2is oxygen-Ci-5 alkylene optionally substituted with one or more selected from the group consisting of C1-5 alkyl, amino, and hydroxide; and R1, R1and R2are independently hydrogen, a linear or branched Ci-36 alkyl, optionally inserted with one or more selected from the group consisting of ester, carbonate, amino, amide, and oxygen, and optionally substituted with one or more selected from the group consisting of halo-Ci-5 alkyl, C1-5 alkyl, amino, hydroxide, and halogen.

[0025] In some embodiments, L1and L2are independently amino-Ci-5 alkylene optionally substituted with one or more selected from the group consisting of alkyl, amino, and hydroxide; and R1, R1, R2, and R2are independently hydrogen, a linear or branched Ci-36 alkyl, optionally inserted with one or more selected from the group consisting of ester, carbonate, amino, amide, and oxygen, and optionally substituted with one or more selected from the group consisting of halo-Ci-5 alkyl, C1-5 alkyl, amino, hydroxide, and halogen.Docket No. 084284.00356

[0026] In another aspect, another subset of compounds of Formula (I) includes the Formula (IH):R3K ZR3"z3R1 / fZR2R1'A BR2' (IH),

[0027] wherein A, B and E are independently a furan or tetrahydrofuran; L1, L2, and L3are independently ester, ether, amide, amino-Ci-5 alkylene, or oxygen-Ci-5 alkylene, wherein amino-C1-5 alkylene and oxygen-Ci-5 alkylene may optionally be substituted with one or more selected from the group consisting of C1-5 alkyl, amino, and hydroxide, and wherein amino-Ci-5 alkylene, or oxygen-Ci-5 alkylene are attached to A, B, and E at alkylene end; R1, R1, R2, R2R3, and R3" are independently hydrogen, a void or linear or branched Ci-36 alkyl, optionally inserted with one or more selected from the group consisting of ester, carbonate, amino, amide, and oxygen, and optionally substituted with one or more selected from the group consisting of halo-Ci-5 alkyl, C1-5 alkyl, amino, hydroxide, and halogen, wherein the void is represented by a dotted line, wherein when R1, R2, or R3is not void, it is connected via a single bond; wherein when L1is amino-Ci-5 alkylene or an amide, L1is bonded via nitrogen to both R1and R1”, wherein one or both of R1or R1can be hydrogen, when L2is amino-Ci-5 alkylene or amide, L2is bonded via nitrogen to both R2and R2”, wherein one or both of R2or R2can be hydrogen, when L3is amino-Ci-5 alkylene or amide, L3is bonded via nitrogen to both R3and R3, wherein one or both of R3or R3can be hydrogen, when L1is ester, ether, or an oxygen-Ci-5 alkylene, only R1is bonded via oxygen to L1and R1is void, when L2is ester, ether, or an oxygen-Ci-5 alkylene, only R2is bonded via oxygen to L2and R2is void, and when L3is ester, ether, or an oxygen-Ci-5 alkylene, only R3is bonded via oxygen to L3and R3is void.Docket No. 084284.00356

[0028] In some embodiments, L1, L2, and L3are independently an amino-Ci-5 alkylene optionally substituted with one or more selected from the group consisting of C1-5 alkyl, amino, and hydroxide; and R1, R1”, R2, R2, R3, and R3are independently hydrogen, a void or a linear or branched Ci-36 alkyl, optionally inserted with one or more selected from the group consisting of ester, carbonate, amino, amide, and oxygen, and optionally substituted with one or more selected from the group consisting of halo-Ci-5 alkyl, C1-5 alkyl, amino, hydroxide, and halogen.

[0029] In another aspect, the present disclosure relates to a lipid nanoparticle composition comprising the lipid compound and a helper lipid, wherein the helper lipid is selected from the group consisting of phospholipid, a structural lipid, pegylated-lipid, and a combination thereof, and the lipid nanoparticle encloses a therapeutic agent.

[0030] In some embodiments, the helper lipid comprises a phospholipid, a structural lipid, pegylated-lipid, or a combination thereof. In some embodiments, the phospholipid is selected from the group consisting of Dilauroyl lecithin (DLPC), Dimyristoyl phosphatidylcholine (DMPC), Dioleoyl lecithin (DOPC), Dipalmitoyl phosphatidylcholine (DPPC), Distearoyl phosphatidylcholine (DSPC), Dioleoyl phosphatidylcholine (DUPC), Palmitoyl oleoyl phosphatidylcholine (POPC), l,2-Di-O-octadecyl-sn-glycero-3-phosphocholine, l-Oleoyl-2-cholesteryldimethylsuccinoyl-sn-glycero-3-phosphocholine, 1-Hexadecyl-sn-gly cero-3 -phosphocholine, 1,2-Divinyl-sn-gly cero-3 -phosphocholine, 1,2-Diarylacyl-sn-glycero-3 -phosphocholine, l,2-Dioleoyl-sn-glycero-3 phosphoryl ethanolamine (DOPE), 1,2-Di-phytanoyl-sn-glycero-3-phosphoethanolamine, 1,2-Distearoyl-sn-glycero-3-phosphoethanolamine, 1,2-Diethenol-sn-gly cero-3 -phosphoethanolamine, 1,2-Divinyl-sn-glycero-3 -phosphoethanolamine, 1,2-Diaryl-sn-gly cero-3 -phosphoethanolamine, 1,2-Dithiohexaenoate-sn-gly cero-3 -Docket No. 084284.00356phosphoethanolamine, l,2-dioleoyl-sn-glycero-3-phosphate-(l -glycerol) sodium salt (DOPG), and combinations thereof.

[0031] In some embodiments, the structural lipid is selected from the group consisting of cholesterol, coprosterol, fecosterol, sitosterol, ergosterol, campesterol, stigmasterol, brassicasterol, tomatidine, tomatine, ursolic acid, alpha-tocopherol, and combinations thereof.

[0032] In some embodiments, the pegylated-lipid is selected from the group consisting of 1,2-dimyristoyl-sn-glycerol methoxypolyethylene glycol (DMG-PEG), 1,2-distearoyl-sn-glycero-3-phosphoethanolamine-N-[amino(polyethylene glycol)] (DSPE-PEG), PEG-disteryl glycerol (DSG-PEG), PEG-dipalmetoleyl, PEG-dioleyl, PEG-distearyl, PEG-diacylglycamide (DAGPEG), PEG-dipalmitoyl phosphatidylethanolamine (DPPE-PEG), or PEG-1,2-dimyristyloxlpropyl-3 -amine (DMA-c-PEG), and combinations thereof.

[0033] In some embodiments, the phospholipid is DOPE, the structural lipid is cholesterol, and the pegylated-lipid is DMG-PEG, wherein a molar ratio of the furan-derived lipid, the DOPE, the cholesterol, and the DMG-PEG is about 20 / 30 / 40 / 0.75. In some embodiments, the lipid nanoparticles further comprise an mRNA encapsulated in the lipid nanoparticle, wherein a mass ratio of the furan-derived lipid to the mRNA is from about 5: 1 to 30: 1. In some embodiments, the mRNA encodes for firefly luciferase (FLuc), green fluorescent protein (GFP), glial cell line-derived neurotrophic factor (GDNF), ere recombinase (Cre), brain-derived neurotrophic factor (BDNF), nerve growth factor (NGF), neurturin (NRTN), insulin-like growth factor-1 (IGF-1), vascular endothelial growth factor (VEGF), aducanumab, donanemab, gantenerumab, solanezumab, crenezumab, etanercept, trastuzumab, natalizumab, briakinumab, bevacizumab, or lecanemab.Docket No. 084284.00356

[0034] In another aspect, the present disclosure relates to a method of intracellular delivery of a therapeutic agent, comprising contacting a cell or administering to a subject a lipid nanoparticle disclosed herein. In some embodiments, the therapeutic agent is selected from the group consisting of mRNA encoding for firefly luciferase (FLuc), green fluorescent protein (GFP), glial cell line-derived neurotrophic factor (GDNF), ere recombinase (Cre), brain-derived neurotrophic factor (BDNF), nerve growth factor (NGF), neurturin (NRTN), insulin-like growth factor-1 (IGF-1), vascular endothelial growth factor (VEGF), aducanumab, donanemab, gantenerumab, solanezumab, crenezumab, etanercept, trastuzumab, natalizumab, briakinumab, bevacizumab, or lecanemab.

[0035] In another aspect, the present disclosure relates to a composition comprising the lipid nanoparticles of the present disclosure.

[0036] Any compositions or methods provided herein can be combined with one or more of any of the other compositions and methods provided herein.BRIEF DESCRIPTION OF THE DRAWINGS

[0037] FIG. 1 is a graph illustrating the luminescence intensity of the signals received from the injection of lipid nanoparticle head groups of the present disclosure.

[0038] FIG. 2 is a graph illustrating the luminescence intensity of the signals received from the injection of lipid tail groups of the present disclosure.

[0039] FIG. 3 is a graph illustrating the luminescence intensity of the signals received from various lipid nanoparticles of the present disclosure.

[0040] FIG. 4 is a graph illustrating the luminescence intensity of the signals received from lipid nanoparticles injected into various organs.Docket No. 084284.00356

[0041] FIG. 5 A is a graph showing the cellular uptake of LNPs encapsulated with Alexa-Fluor 647-labeled RNA in various embodiments of LNPs.

[0042] FIG. 5B is a graph showing the cellular uptake of LNPs encapsulated with Alexa-Fluor 647-labeled RNA in various embodiments of LNPs.

[0043] FIG. 6A is an immunofluorescence flow cytometry (IFCM) graph of GFP expression in different brain cell types from the brains of the mice subcutaneously injected with various embodiments of LNPs.

[0044] FIG. 6B is another immunofluorescence flow cytometry (IFCM) graph of GFP expression in different brain cell types from the brains of the mice subcutaneously injected with various embodiments of LNPs.

[0045] FIG. 7A is a graph showing the blood urea nitrogen (BUN) level evaluations of the mice after subcutaneous administration of F6T1, Fl 1T6, or SM102 LNPs. ns = not significant.

[0046] FIG. 7B is a graph showing the alanine transaminase (ALT) level evaluation of the mice after subcutaneous administration of F6T1, Fl 1T6, or SM102 LNPs. ns = not significant.

[0047] FIG. 7C is a graph showing the aspartate aminotransferase (AST) evaluation of the mice after subcutaneous administration of F6T1, F7T1, or SM102 LNPs. ns = not significant.

[0048] FIG. 8A is a graph showing the tdTomato expression of various LNPs in neurons.

[0049] FIG. 8B is a graph showing the tdTomato expression of various LNPs in microglia.

[0050] FIG. 8C is a graph showing the tdTomato expression of various LNPs in astrocytes.DETAILED DESCRIPTION

[0051] The following discussion omits or only briefly describes conventional features of the disclosed technology that are apparent to those skilled in the art. Reference to various embodiments does not limit the scope of the claims attached hereto. Additionally, any examplesDocket No. 084284.00356set forth in this specification are intended to be non-limiting and merely set forth some of the many possible embodiments for the appended claims. Further, particular features described herein can be used in combination with other described features in each of the various possible combinations and permutations. A person of ordinary skill in the art would know how to make and use the disclosed technology, in combination with routine experiments, to achieve other outcomes not specifically disclosed in the examples or the embodiments.

[0052] Unless defined otherwise, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which the disclosure pertains. Although any methods and materials similar or equivalent to those described herein can be used in the practice for testing of the present disclosure, the preferred materials and methods are described herein. In describing and claiming the present disclosure, the following terminology will be used. It is also to be understood that the terminology used herein is for the purpose of describing particular embodiments only, and is not intended to be limiting.

[0053] The articles “a” and “an” are used herein to refer to one or to more than one (i.e., to at least one) of the grammatical object of the article. By way of example, “an element” means one element or more than one element. Thus, recitation of “a cell”, for example, includes a plurality of cells of the same type. Furthermore, to the extent that the terms “including”, “includes”, “having”, “has”, “with”, or variants thereof are used in either the detailed description and / or the claims, such terms are intended to be inclusive in a manner similar to the term “comprising.”

[0054] ‘About” as used herein when referring to a measurable value such as an amount, a temporal duration, and the like, is meant to encompass variations of + / - 20%, + / - 10%, + / - 5%, + / -1%, or + / - 0.1% from the specified value, as such variations are appropriate to perform the disclosed methods. Alternatively, particularly with respect to biological systems or processes, theDocket No. 084284.00356term can mean within an order of magnitude within 5-fold, and also within 2-fold, of a value. Where particular values are described in the application and claims, unless otherwise stated the term “about” meaning within an acceptable error range for the particular value should be assumed.

[0055] As used herein, the terms “agent” or “therapeutic agent” are meant to encompass any molecule, chemical entity, composition, drug, therapeutic agent, chemotherapeutic agent, or biological agent capable of preventing, ameliorating, or treating a disease or other medical condition. The term includes small molecule compounds, antisense reagents, mRNA, siRNA reagents, antibodies, antibody fragments bearing epitope recognition sites, such as Fab, Fab’, F(ab’)2 fragments, Fv fragments, single chain antibodies, antibody mimetics (such as DARPins, affibody molecules, affilins, affitins, anticalins, avimers, fynomers, Kunitz domain peptides and monobodies), peptoids, aptamers; hormones, oligonucleotides, enzymes, peptides organic or inorganic molecules, natural or synthetic compounds and the like. An agent can be assayed in accordance with the methods of the disclosure at any stage during clinical trials, during pre-trial testing, or following FDA-approval.

[0056] As used herein, the terms “comprising,” “comprise” or “comprised,” and variations thereof, in reference to defined or described elements of an item, composition, apparatus, method, process, system, etc. are meant to be inclusive or open-ended, permitting additional elements, thereby indicating that the defined or described item, composition, apparatus, method, process, system, etc. include those specified elements— or, as appropriate, equivalents thereof— and that other elements can be included and still fall within the scope / definition of the defined item, composition, apparatus, method, process, system, etc.

[0057] An “effective amount” as used herein, means an amount that provides a therapeutic or prophylactic benefit.Docket No. 084284.00356

[0058] The term “encapsulation efficiency” as used herein refers to the percentage of nucleic acid in the lipid nanoparticles that is not degraded after exposure to serum or a nuclease assay that would significantly degrade free nucleic acids. As used herein, “encapsulation” may refer to complete, substantial, or partial enclosure, confinement, surrounding, or encasement.

[0059] The term “fully encapsulated” as used herein indicates that the mRNA in the nanoparticles is not significantly degraded after exposure to serum or a nuclease assay that would significantly degrade free nucleic acids. In a fully encapsulated system, preferably less than 25% of particle nucleic acid is degraded in a treatment that would normally degrade 100% of free nucleic acid, more preferably less than 10%, and most preferably less than 5% of the particle nucleic acid is degraded. Fully encapsulated also indicates that the particles are serum stable, that is, that they do not rapidly decompose into their component parts upon in vivo administration.

[0060] As used herein, the term “in vitro” refers to events that occur in an artificial environment, e.g., in a test tube or reaction vessel, in cell culture, in a Petri dish, etc., rather than within an organism (e.g., animal, plant, or microbe).

[0061] As used herein, the term “in vivo” refers to events that occur within an organism (e.g., animal, plant, or microbe or cell or tissue thereof).

[0062] The term “lipid nanoparticle” refers to a nanoparticle that includes lipids and that is stable and dispersible in aqueous media. As used herein, the term “nanoparticle” refers to a particle having one or a plurality of components, the particle having any one structural feature on a scale of less than about 1000 nm that exhibits novel properties as compared to a bulk sample of the same material or component materials. Routinely, nanoparticles have any one structural feature on a scale of less than about 500 nm, less than about 400 nm, less than about 300 nm, less than about 200 nm, or less than about 100 nm. In exemplary embodiments, a nanoparticle is a particleDocket No. 084284.00356having one or more dimensions of the order of about 10 nm to about 500 nm. In other exemplary embodiments, a nanoparticle is a particle having one or more dimensions of the order of about 10 nm to about 1000 nm. A spherical nanoparticle would have a diameter, for example, of between 10 nm to about 100 nm or 10 nm to about 1000 nm.

[0063] A nanoparticle most often behaves as a unit in terms of its physical or biophysical properties, e.g., transport. It is noted that novel properties that differentiate nanoparticles from the corresponding bulk material typically develop at a size scale of under 1000 nm, or at a size of under 500 nm, but nanoparticles can be of a larger size, for example, for particles that are oblong, tubular, and the like. The size at which materials display different properties as compared to the bulk material is material-dependent and can be seen for many materials much larger in size than 100 nm and even for some materials larger in size than 1000 nm. Nanoparticles can be employed in a variety of drug delivery technologies (e.g., mRNA delivery technologies) and can be employed for various purposes including, but not limited to, controlled drug delivery, protection of the drugs from degradation, and protection of the body from the toxic effects of the drugs.

[0064] As used in this specification and the appended claims, the term “or” is generally employed in its sense including “and / or” unless the content clearly dictates otherwise.

[0065] “Parenteral” administration of an immunogenic composition includes, e.g., subcutaneous, intravenous, intramuscular, intrathecal, or intrasternal injection, or infusion techniques.

[0066] The terms “patient” or “individual” or “subject” are used interchangeably herein, and refers to a mammalian subject to be treated, with human patients being preferred. In some cases, the methods of the disclosure find use in experimental animals, in veterinary application,Docket No. 084284.00356and in the development of animal models for disease, including, but not limited to, rodents including mice, rats, hamsters, and primates.

[0067] As used herein, a “pharmaceutically acceptable” component / carrier etc. is one that is suitable for use with humans and / or animals without undue adverse side effects (such as toxicity, irritation, and allergic response) commensurate with a reasonable benefit / risk ratio.

[0068] Treatment” is an intervention performed with the intention of preventing the development or altering of the pathology or symptoms of a disorder. Accordingly, “treatment” refers to both therapeutic treatment and prophylactic or preventative measures. “Treatment” may also be specified as palliative care. Those in need of treatment include those already with the disorder as well as those in which the disorder is to be prevented. As defined herein, a “therapeutically effective” amount of a compound or agent (i.e., an effective dosage) means an amount sufficient to produce a therapeutically (e.g., clinically) desirable result. The compositions can be administered from one or more times per day to one or more times per week; including once every other day. The skilled artisan will appreciate that certain factors can influence the dosage and timing required to effectively treat a subject, including but not limited to the severity of the disease or disorder, previous treatments, the general health and / or age of the subject, and other diseases present. Moreover, the treatment of a subject with a therapeutically effective amount of the compounds of the disclosure can include a single treatment or a series of treatments.

[0069] Ranges: throughout this disclosure, various aspects of the disclosure can be presented in a range format. It should be understood that the description in range format is merely for convenience and brevity and should not be construed as an inflexible limitation on the scope of the disclosure. Accordingly, the description of a range should be considered to have specifically disclosed all the possible subranges as well as individual numerical values within that range. ForDocket No. 084284.00356example, a description of a range such as from 1 to 6 should be considered to have specifically disclosed subranges such as from 1 to 3, from 1 to 4, from 1 to 5, from 2 to 4, from 2 to 6, from 3 to 6 etc., as well as individual numbers within that range, for example, 1, 2, 2.7, 3, 4, 5, 5.3, and 6. This applies regardless of the breadth of the range.

[0070] Delivery of messenger RNA (mRNA) to the central nervous system (CNS) remains a significant challenge. The present document provides a library of furan-derived ionizable lipid nanoparticles that leverage the meningeal lymphatic vessel (MLVs) route to achieve efficient mRNA delivery to the brain. These ionizable lipids are engineered with different ring structures, e.g., furan or tetrahydrofuran cores, functional groups, and tails. These furan and tetrahydrofuran-containing lipid nanoparticles (LNPs) generally displayed exceptional mRNA delivery. These LNPs demonstrated significantly higher mRNA delivery efficiency to the brain compared to FDA-approved SM102 LNPs. Confocal imaging revealed that these LNPs bypassed the blood-brain barrier (BBB) via the lymphatic pathway, traveling from the deep cervical lymph nodes (dCLNs) to the meninges and subsequently entering brain cells.

[0071] In contrast to liposomes and cationic liposomes, lipid nanoparticles have a structure that includes a single monolayer or bilayer of lipids that encapsulates a compound in a solid phase. Thus, unlike liposomes, lipid nanoparticles do not have an aqueous phase or other liquid phase in their interior, but rather the lipids from the bilayer or monolayer shell are directly complexed to the internal compound thereby encapsulating it in a solid core. Lipid nanoparticles are typically spherical vesicles having a relatively uniform dispersion of shape and size ranging from about 10 nm to 1000 nm.

[0072] In the disclosed lipid nanoparticle mRNA delivery systems, the lipid is formulated to include a furan-derived ionizable lipid which can complex to and associate with the negativelyDocket No. 084284.00356charged backbone of the mRNA. Furan-derived ionizable lipids with apparent pKa values below 7 have the benefit of providing a cationic environment for complexing with the negative charge on the mRNA acid and loading into the lipid nanoparticle at pH values below the pKa of the furanderived ionizable lipid where it is positively charged. Then, at physiological pH values, e.g., pH between about 7.35 to about 7.45, the lipid nanoparticle can adopt a relatively neutral exterior allowing for a significant increase in the lifetime of the lipid particles following administration. In the context of mRNA delivery, lipid nanoparticles offer many advantages over other lipid-based nucleic acid delivery systems including greater encapsulation efficiency, potent transfection, improved delivery efficiency, and low levels of cytotoxicity and immunogenicity.

[0073] In the context of the present disclosure, a lipid nanoparticle (LNP) delivery vehicle typically serves to transport a desired therapeutic agent to a target cell or tissue. In certain embodiments, the lipid formulation encapsulates the therapeutic agent.Ionizable Lipid

[0074] In one aspect, the lipid nanoparticle formulation comprises an ionizable lipid, wherein the ionizable lipid comprises a head group and a hydrophobic tail group. In some embodiments, the head group may comprise a ring structure including but not limited to a furan, tetrahydrofuran, or pyridine. In some embodiments, the ring structure may include any heterocyclic group or carbocyclic group including cycloalkyl or aryl groups. In some embodiments, the head group may comprise a core comprising one or more ring structures optionally inserted with one or more heteroatoms including N, O, or S. In some embodiments, any one of ring structure disclosed herein may be optionally substituted with a linear or branched Ci-is alkyl, Ci-isalkoxyl-alkylene, hydrogen, halo-Ci-5 alkyl, C1-5 alkyl, C1-6 cycloalkyl-Ci-is alkylene, amino, hydroxide, or halogen:Docket No. 084284.00356

[0075] In some embodiments, carbocycles may include cyclopropyl, cyclopentyl, cyclohexyl, phenyl, naphthyl, and 1,2-dihydronaphthyl groups.

[0076] In some embodiments, the ring structure may include heterocycloalkyl or heteroaryl groups including but not limited to include imidazolyl, imidazolidinyl, oxazolyl, oxazolidinyl, thiazolyl, thiazolidinyl, pyrazolidinyl, pyrazolyl, isoxazolidinyl, isoxazolyl, isothiazolidinyl, isothiazolyl, morpholinyl, pyrrolyl, pyrrolidinyl, furyl, tetrahydrofuryl, thiophenyl, pyridinyl, piperidinyl, quinolyl, and isoquinolyl groups.

[0077] As used herein, the term “carbocycle” or “carbocyclic group” means an optionally substituted mono- or multi-cyclic system including one or more rings of carbon atoms. Rings may be three, four, five, six, seven, eight, nine, ten, eleven, twelve, thirteen, fourteen, fifteen, sixteen, seventeen, eighteen, nineteen, or twenty membered rings. The notation “C3-6 carbocycle” means a carbocycle including a single ring having 3-6 carbon atoms. Carbocycles may include one or more carbon-carbon double or triple bonds and may be non-aromatic or aromatic (e.g., cycloalkyl or aryl groups). Examples of carbocycles include cyclopropyl, cyclopentyl, cyclohexyl, phenyl, naphthyl, and 1,2-dihydronaphthyl groups. The term “cycloalkyl” as used herein means a non-aromatic carbocycle and may or may not include any double or triple bond. Unless otherwise specified, carbocycles described herein refers to both unsubstituted and substituted carbocycle groups, i.e., optionally substituted carbocycles.

[0078] As used herein, the term “heterocycle” or “heterocyclic group” means an optionally substituted mono- or multi-cyclic system including one or more rings, where at least one ring includes at least one heteroatom. Heteroatoms may be, for example, nitrogen, oxygen, or sulfur atoms. Rings may be three, four, five, six, seven, eight, nine, ten, eleven, twelve, thirteen, or fourteen membered rings. Heterocycles may include one or more double or triple bonds and mayDocket No. 084284.00356be non-aromatic or aromatic (e.g., heterocycloalkyl or heteroaryl groups). Examples of heterocycles include imidazolyl, imidazolidinyl, oxazolyl, oxazolidinyl, thiazolyl, thiazolidinyl, pyrazolidinyl, pyrazolyl, isoxazolidinyl, isoxazolyl, isothiazolidinyl, isothiazolyl, morpholinyl, pyrrolyl, pyrrolidinyl, furyl, tetrahydrofuryl, thiophenyl, pyridinyl, piperidinyl, quinolyl, and isoquinolyl groups. The term “heterocycloalkyl” as used herein means a non-aromatic heterocycle and may or may not include any double or triple bond. Unless otherwise specified, heterocycles described herein refers to both unsubstituted and substituted heterocycle groups, i.e., optionally substituted heterocycles.

[0079] In some embodiments, the ring structure may comprise a furan or a tetrahydrofuran (THF). As used herein, the term “furan-derived” refers to a structure that contains a furan moiety, or a derivative of furan including but not limited to tetrahydrofuran, furfural, 5-hydroxymethylfurfural, furanocoumarins, furanones, or dibenzofuran. As used in this document, “furan-derived” means it contains a head group that contains one or more furan or tetrahydrofuran. In this document “furan-derived lipid” and “furan-derived ionizable lipids” are used interchangeably. The core furan or tetrahydrofuran moiety of a lipid according to Formula (I) may be referred to as cationic or ionizable lipids. Lipids may also be zwitterionic, i.e., neutral molecules having both a positive and a negative charge.

[0080] As used herein, the term “alkyl” refers to a group containing one or more carbon atoms (for example, one, two, three, four, five, six, seven, eight, nine, ten, eleven, twelve, thirteen, fourteen, fifteen, sixteen, seventeen, eighteen, nineteen, twenty or more carbon atoms), which may be optionally substituted. For instance, the term “Ci-36 alkyl” refers to an optionally substituted linear or branched saturated hydrocarbon containing from 1 to 36 carbon atoms, for example, 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30,Docket No. 084284.0035631, 32, 33, 34, 35, or 36. Unless otherwise explicitly indicated, alkyl groups described herein refer to both unsubstituted and substituted alkyl groups.

[0081] As used herein, the term “alkylene” refers to a straight, branched chain, or cyclic structure of chain divalent, trivalent, or tetraval ent hydrocarbon radical having from 1 to 36 carbon atoms, optionally substituted with substituents selected from the group consisting of lower alkyl, lower alkoxy, lower alkylsulfanyl, lower alkylsulfenyl, lower alkylsulfonyl, oxo, hydroxy, mercapto, amino optionally substituted by alkyl, carboxy, carbamoyl optionally substituted by alkyl, aminosulfonyl optionally substituted by alkyl, silyloxy optionally substituted by alkoxy, alkyl, or aryl, silyl optionally substituted by alkoxy, alkyl, or aryl, nitro, cyano, halogen, or lower perfluoroalkyl, multiple degrees of substitution being allowed. Such an “alkylene” group may contain one or more O, S, S(O), or S(O)2 atoms. Examples of “alkylene” as used herein include, but are not limited to, methylene, ethylene, propylene, ethan- 1,1 -diyl, ethan-l,2-diyl, propan- 1,1-diyl, propan- 1,3 -diyl, propan-1, 2-diyl, butan-l,4-diyl, butan- 1,3 -diyl, butan-l,2-diyl, 2-methyl-propan- 1,3 -diyl, and the like. Unless otherwise explicitly indicated, alkylene groups described herein refer to both unsubstituted and substituted alkylene groups.

[0082] As used herein, “cycloalkyl” refers to an alicyclic hydrocarbon group optionally possessing one or more degrees of unsaturation, having from three to twelve carbon atoms, optionally substituted with substituents selected from the group consisting of lower alkyl, lower alkoxy, lower alkylsulfanyl, lower alkylsulfenyl, lower alkylsulfonyl, oxo, hydroxy, mercapto, amino optionally substituted by alkyl, carboxy, carbamoyl optionally substituted by alkyl, aminosulfonyl optionally substituted by alkyl, nitro, cyano, halogen, or lower perfluoroalkyl, multiple degrees of substitution being allowed. “Cycloalkyl” includes by way of example cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, cycloheptyl, or cyclooctyl, and the like. UnlessDocket No. 084284.00356otherwise explicitly indicated, cycloalkyl groups described herein refer to both unsubstituted and substituted cycloalkyl groups.

[0083] As used herein, “halide,” “halo,” or “halogen” refers to a chloro, bromo, fluoro, or iodo atom radicals. Similarly, as used herein, “haloalkyl” means a hydrocarbon substituent, which is a linear or branched, alkyl, alkenyl, or alkynyl substituted with one or more chloro, bromo, fluoro, and / or iodo atom(s). Similarly, as used herein, “halo-Cn-malkyl” or “Cn-m haloalkyl” refers to an alkyl group having from one halogen atom to 2s+ 1 halogen atoms which may be the same or different, where “s” is the number of carbon atoms in the alkyl group, wherein the alkyl group has n to m carbon atoms. In some embodiments, the haloalkyl group is fluorinated only. In some embodiments, the alkyl group has 1, 2, 3, 4, or 5 carbon atoms.

[0084] As disclosed throughout this document, all stereoisomers of the compounds are encompassed by the foregoing formulas. When no particular stereoisomer or stereochemistry is indicated, it is understood to mean all possible stereoisomers that could be produced from a reaction are present. A person of ordinary skill in the art will recognize that the reactions can be optimized to give one isomer preferentially, or new schemes may be devised to produce a single isomer.

[0085] In one aspect, the Furan-derived ionizable lipid compounds are described herein having Formula (I):R1'L, R2"R1 A BR2' (I)

[0086] wherein A is a heteroaryl, aryl, cycloalkyl, heterocycloalkyl, a furan or tetrahydrofuran; B is a bond, a heteroaryl, aryl, cycloalkyl, heterocycloalkyl, a furan or tetrahydrofuran, wherein A and B are optionally and independently substituted at any of the carbonDocket No. 084284.00356positions with a linear or branched Ci-is alkyl, Cuis alkoxyl-alkylene, hydrogen, halo-Ci-5 alkyl, Ci-5 alkyl, amino, hydroxide, or halogen; L1and L2are independently ester, ether, amide, amino-C1-5 alkylene, or oxygen-Ci-5 alkylene, wherein amino-Ci-5 alkylene and oxygen-Ci-5 alkylene may be optionally substituted with one or more of C1-5 alkyl, amino, and hydroxide, and wherein amino-C1-5 alkylene or oxygen-Ci-5 alkylene are attached to A or B at alkylene end; R1, R1”, R2and R2are independently hydrogen, a void or a linear or branched Ci-36 alkyl, optionally inserted with one or more ester, carbonate (-O-(C=O)-O-), amino (-NH2), amide, or oxygen (-O-), and optionally substituted with one or more of halo-Ci-5 alkyl, C1-5 alkyl, amino, hydroxide, or halogen; wherein

[0087] when L1is amino-Ci-5 alkylene or an amide, L1is bonded via nitrogen to both R1and R1”, wherein one or both of R1or R1can be hydrogen, when L1is ester, ether, or an oxygen-C1-5 alkylene, only R1is bonded via oxygen to L1and R1is void, when L2is amino-Ci-5 alkylene or amide, L2is bonded via nitrogen to both R2and R2, wherein one or both of R2or R2can be hydrogen, when L2is ester, ether, or an oxygen-Ci-5 alkylene, only R2is bonded via oxygen to L2and R2is void,

[0088] V is a bond, a linear or branched C1-5 alkylene optionally substituted with one or more C1-5 alkyl, (-[C(L4)(L5)]-), C3-6 cycloalkyl, or C1-5 alkylene-D, wherein D is E-L3(R3)(R3); E is a heteroaryl, aryl, cycloalkyl, heterocycloalkyl, a furan or tetrahydrofuran optionally and independently substituted at any of the carbon positions with a linear or branched Ci-is alkyl, Ci-18 alkoxyl-alkylene, hydrogen, halo-Ci-5 alkyl, C1-5 alkyl, amino, hydroxide, or halogen; L3is an ester, ether, amide, amino-Ci-5 alkylene, or oxygen-Ci-5 alkylene, wherein amino-Ci-5 alkylene and oxygen-Ci-5 alkylene may optionally be substituted with one or more of Ci 5 alkyl, amino, or hydroxide, and wherein amino-Ci-5 alkylene or oxygen-Ci-5 alkylene are attached to E at the alkylene end; when L3is amino-Ci-5 alkylene or amide, L3 is bonded via nitrogen to both R3andDocket No. 084284.00356R3, wherein one or both of R3or R3can be hydrogen, when L3is ester, ether, or an oxygen-Ci-5 alkylene, only R3is bonded via oxygen to L3and R3is void; R3and R3are independently hydrogen, a void or a linear or branched Ci-36 alkyl, optionally inserted with one or more ester, carbonate (-O-(C=O)-O-), amino (-NH2), amide, or oxygen (-O-), and optionally substituted with one or more of halo-Ci-5 alkyl, C1-5 alkyl, amino, hydroxide, or halogen. In some embodiments, at least one of L1, L2, or L3is amino-Ci-5 alkylene.

[0089] As used herein, the term “inserted” refers to an element or a functional group that is positioned between any two carbon atoms or at a terminus of an alkyl or alkylene chain. In this configuration, the numbering of carbon atoms, (i.e., Cl, C2, C3... etc.) only refers to the carbon atom along the alkyl or alkylene chain and does not include any heteroatoms (i.e., oxygen, nitrogen, etc.) from the insertion group.

[0090] In some embodiments, A is connected to L1at ring position 2 of A, i.e., furan or tetrahydrofuran, and A is connected to V at ring position 5 of A, based on the ring numbering shown below, wherein the dotted line represents potential double bond together with an existing bond, or void;1\» / /

[0091] As seen in Formula (I) and used herein, the furan-derived ionizable lipid compounds include a “head group” defined as -[L'-A-V-B-L2]- and any subset of compounds described in this document. In some embodiments, the furan-derived ionizable lipid compounds include a “tail group” defined as any one of R1, R1, R2, R2, R3, and R3and any subset of compounds described in this document. In some embodiments, a tail group (i.e., R1, R1”, R2, R2,Docket No. 084284.00356R3, and R3) may be attached to the head group at L1, L2, or L3using methods commonly known in the art. For example, when L1, L2, or L3, of the head group contains an amino or an amide, a tail group may be installed via epoxide ring-opening reactions or reductive amination reactions. In some embodiments, when L1, L2, or L3, of the head group contains an ester, ether, or oxygen, a tail group may be installed through various alkylation reactions.

[0092] In some embodiments, a tail group may comprise a Ci-36 alkyl chain inserted with one or more carbonate moiety, an ester moiety, an amino moiety, an amide moiety, or oxygen along the Ci-36 alkyl chain. In some embodiments, a tail group may comprise a Ci-36 alkyl chain inserted with two oxygen atoms between one, two, or three carbon atoms. In some embodiments, the two oxygens are positioned between a single carbon atom forming an acetal ether linkage.

[0093] In certain embodiments, a subset of compounds of Formula (I) includes those of Formula (IA):

[0094] wherein A is a heteroaryl, aryl, cycloalkyl, heterocycloalkyl, a furan or tetrahydrofuran, optionally substituted at any of the carbon positions with a linear or branched Ci-18 alkyl, Cnisalkoxyl-alkylene, hydrogen, halo-Ci-s alkyl, C1-5 alkyl, amino, hydroxide, or halogen;

[0095] V and B of Formula (I) are both a bond, wherein when V and B are both a bond, there is one bond between A and L2; L1and L2are independently ester, ether, amide, amino-Ci-5 alkylene, or oxygen-Ci-s alkylene, wherein amino-Ci-5 alkylene and oxygen-Ci-5 alkylene may beDocket No. 084284.00356optionally substituted with one or more of C1-5 alkyl, amino, and hydroxide, and wherein amino-C1-5 alkylene or oxygen-Ci-5 alkylene are attached to A at the alkylene end;

[0096] R1, R1”, R2and R2are independently hydrogen, a void (represented by dotted line) or a linear or branched Ci-36 alkyl, optionally inserted with one or more ester, carbonate (-0-(C=O)-O-), amino (-NH2), amide, or oxygen (-O-), and optionally substituted with one or more of halo-Ci-5 alkyl, C1-5 alkyl, amino, hydroxide, or halogen; wherein when L1is amino-Ci-5 alkylene or amide, L1is bonded via nitrogen to both R1and R1, wherein one or both of R1or R1can be hydrogen, when L1is ester, ether, or an oxygen-Ci-5 alkylene, only R1is bonded via oxygen to L1and R1is void, when L2is amino-Ci-5 alkylene or amide, L2is bonded via nitrogen to both R2and R2”, wherein one or both of R2or R2can be hydrogen. In this configuration, A is connected to L1at ring position 2 of A, and A is connected to L2at ring position 5 of A. In some embodiments, at least one of L1or L2is amino-Ci-5 alkylene. In some embodiments, when the dotted line is void, the group it is attached to is also void. In some embodiments, the dotted line is a void or a single bond.

[0097] In certain embodiments, a subset of compounds of Formula (I) or Formula (IA) may include the following Formula (IB):R1"_ Ci-4 Ci-4K(IB)wherein L1from Formula (I) or Formula (IA) is amino-Ci-5 alkylene optionally substituted with one or more of C1-5 alkyl, amino, hydroxide; L2from Formula (I) or Formula (IA) is oxygen-Ci-5 alkylene optionally substituted with one or more of C1-5 alkyl, amino, or hydroxide.Docket No. 084284.00356

[0098] In some embodiments, L1may be amino-Ci alkylene, amino-C2 alkylene, amino-C3 alkylene, amino-C4 alkylene, or amino-Cs alkylene, optionally substituted with Ci 5 alkyl, including but not limited to Ci alkyl, C2 alkyl, C3 alkyl, C4 alkyl, or C5 alkyl. In some embodiments, L1may be substituted with two C1-5 alkyl, to form, for example, a gemdimethyl-alkyl. In some embodiments, L2may be oxygen-Ci alkylene, oxygen-C2 alkylene, oxygen-Cs alkylene, oxygen-C4 alkylene, or oxygen-Cs alkylene, optionally substituted with C1-5 alkyl, including but not limited to Ci alkyl, C2 alkyl, C3 alkyl, C4 alkyl, or Cs alkyl. In some embodiments, L2may be substituted with two C1-5 alkyl, to form, for example, a gemdimethyl-alkyl.

[0099] In some embodiments, R1, R1and R2are independently a linear or branched Ci-36 alkyl, optionally inserted with one or more ester, carbonate (-O-(C=O)-O-), amino (-NH2), amide, or oxygen (-O-), and optionally substituted with one or more of hydrogen, halo-Ci-5 alkyl, C1-5 alkyl, amino, hydroxide, or halogen.

[0100] In certain embodiments, a subset of compounds of Formula (I) or Formula (IA) includes those of Formula (IC):R1< A X'R2"CM-4 C1 4K(ic)

[0101] wherein L1and L2from Formula (I) or Formula (IA) are independently amino-Ci-5 alkylene optionally substituted with one or more of C1-5 alkyl, amino, or hydroxide; R1, R1, R2and R2are independently hydrogen, a linear or branched Ci-36 alkyl, optionally inserted with one or more ester, carbonate (-O-(C=O)-O-), amino (-NH2), amide, or oxygen (-O-), and optionally substituted with one or more of halo-Ci-5 alkyl, C1-5 alkyl, amino, hydroxide, or halogen.Docket No. 084284.00356

[0102] In certain embodiments, a subset of compounds of Formula (I) includes those of Formula (ID):

[0103] wherein A and B are independently a heteroaryl, aryl, cycloalkyl, heterocycloalkyl, a furan or tetrahydrofuran, wherein A and B are optionally and independently substituted at any of the carbon positions with a linear or branched Ci-is alkyl, Ci-is alkoxyl-alkylene, hydrogen, halo-C1-5 alkyl, Ci-5 alkyl, amino, hydroxide, or halogen;

[0104] V from Formula (I) is Ci alkylene optionally substituted with one or two C1-5 alkyl, (-[C(L4)(L5)]-); L1and L2are independently ester, ether, amide, amino-Ci-5 alkylene, or oxygen-C1-5 alkylene, wherein amino-Ci-5 alkylene, oxygen-Ci-5 alkylene may be optionally substituted with one or more of C1-5 alkyl, amino, or hydroxide; L4and L5are independently selected from a hydrogen or a C1-5 alkyl; and R1, R1, R2and R2are independently hydrogen, a void (represented by dotted line) or a linear or branched Ci-36 alkyl, optionally inserted with one or more ester, carbonate (-O-(C=O)-O-), amino (-NH2), mono- or di-alkylated amino, amide, or oxygen (-O-), and optionally substituted with one or more of halo-Ci-5 alkyl, C1-5 alkyl, amino, hydroxide, or halogen,

[0105] wherein when L1is amino-Ci-5 alkylene or an amide, L1is bonded via nitrogen to both R1and R1”, wherein one or both of R1or R1can be hydrogen, when L1is ester, ether, or an oxygen-Ci-5 alkylene, only R1is bonded via oxygen to L1and R1is void, when L2is amino-Ci-5 alkylene or amide, L2is bonded via nitrogen to both R2and R2, wherein one or both of R2or R2canbe hydrogen, when L2is ester, ether, or an oxygen-Ci-5 alkylene, only R2is bonded via oxygenDocket No. 084284.00356to L2and R2is void. In some embodiments, when the dotted line is void, the group it is attached to is also void. In some embodiments, the dotted line is a void or a single bond.

[0106] In certain embodiments, a subset of compounds of Formula (I) or Formula (ID) includes those of Formula (IE):

[0107] wherein L1and L2from Formula (I) and Formula (ID) are each amino-Ci-5 alkylene optionally substituted with one or more of C1-5 alkyl, amino, or hydroxide; L4and L5are independently selected from a hydrogen or a C1-5 alkyl; R1, R1, R2and R2are independently hydrogen, a linear or branched Ci-36 alkyl, optionally inserted with one or more ester, carbonate (-O-(C=O)-O-), amino (-NH2), amide, or oxygen (-O-), and optionally substituted with one or more of halo-Ci-5 alkyl, C1-5 alkyl, amino, hydroxide, or halogen.

[0108] In certain embodiments, a subset of compounds of Formula (I) includes those of Formula (IF):Rl", R2'R1' A— B R2' / .p.

[0109] wherein V from Formula (I) is a bond connecting A and B, and A and B are independently a heteroaryl, aryl, cycloalkyl, heterocycloalkyl, a furan or tetrahydrofuran, wherein A and B are optionally and independently substituted at any of the carbon positions with a linear or branched Ci-18 alkyl, Ci-18 alkoxyl-alkylene, hydrogen, halo-Ci-5 alkyl, C1-5 alkyl, amino, hydroxide, or halogen; L1and L2are independently ester, ether, amide, amino-Ci-5 alkylene, or oxygen-Ci-5 alkylene, wherein amino-Ci-5 alkylene and oxygen-Ci-5 alkylene may be optionallyDocket No. 084284.00356substituted with one or more of C1-5 alkyl, amino, or hydroxide; and R1, R1, R2and R2are independently hydrogen, a void (represented by dotted line) or a linear or branched Ci-36 alkyl, optionally inserted with one or more ester, carbonate (-O-(C=O)-O-), amino (-NH2), amide, or oxygen (-O-), and optionally substituted with one or more of halo-Ci-5 alkyl, C1-5 alkyl, amino, hydroxide, or halogen, wherein when L1is amino-Ci-5 alkylene or an amide, L1is bonded via nitrogen to both R1and R1, wherein one or both of R1or R1can be hydrogen, when L1is ester, ether, or an oxygen-Ci-5 alkylene, only R1is bonded via oxygen to L1and R1is void, when L2is amino-Ci-5 alkylene or amide, L2is bonded via nitrogen to both R2and R2, wherein one or both of R2or R2can be hydrogen, when L2is ester, ether, or an oxygen-Ci-5 alkylene, only R2is bonded via oxygen to L2and R2is void. In some embodiments, when the dotted line is void, the group it is attached to is also void. In some embodiments, the dotted line is a void or a single bond.

[0110] In this configuration, A is connected to B at ring position 5 of A and ring position 2 of B, A is connected to L1at ring position 2 of A, and B is connected to L2at ring position 5 of B.

[0111] In certain embodiments, a subset of compounds of Formula (I) and Formula (IF) includes those of Formula (IG):

[0112] wherein A and B are independently a heteroaryl, aryl, cycloalkyl, heterocycloalkyl, a furan or tetrahydrofuran, wherein A and B are optionally and independently substituted at any of the carbon positions with a linear or branched Ci-is alkyl, Ci-18 alkoxyl-alkylene, hydrogen, halo-C1-5 alkyl, C1-5 alkyl, amino, hydroxide, or halogen; L1and L2from Formula (I) and Formula (IF) are each amino-Ci-5 alkylene optionally substituted with one or more of Ci 5 alkyl, amino, orDocket No. 084284.00356hydroxide; R1, R1, R2and R2are independently hydrogen, a linear or branched Ci-36 alkyl, optionally inserted with one or more ester, carbonate (-O-(C=O)-O-), amino (-NH2), amide, or oxygen (-O-), and optionally substituted with one or more of halo-Ci-5 alkyl, C1-5 alkyl, amino, hydroxide, or halogen.

[0113] In certain embodiments, a subset of compounds of Formula (I) includes those of Formula (IH):A2B

[0114] wherein A, B, and E are independently a heteroaryl, aryl, cycloalkyl, heterocycloalkyl, a furan or tetrahydrofuran, wherein A, B, and E are optionally and independently substituted at any of the carbon positions with a linear or branched Ci-is alkyl, Ci-18 alkoxyl-alkylene, hydrogen, halo-Ci-5 alkyl, C1-5 alkyl, amino, hydroxide, or halogen; V from Formula (I) is Ci alkylene-D, and D is E-L3(R3)(R3”); L1, L2, and L3are independently ester, ether, amide, amino-Ci-5 alkylene, or oxygen-Ci-5 alkylene, wherein amino-Ci-5 alkylene and oxygen-Ci-5 alkylene may optionally be substituted with one or more of C1-5 alkyl, hydrogen, amino, or hydroxide, and wherein amino-Ci-5 alkylene or oxygen-Ci-5 alkylene are attached to A, B, and E at the alkylene end; R1, R1, R2, R2, R3and R3are independently hydrogen, a void (represented by dotted line) or linear or branched Ci-36 alkyl, optionally inserted with one or more ester, carbonate (-O-(C=O)-O-), amino (-NH2), amide, or oxygen (-O-), and optionally substituted with one or more of halo-Ci-5 alkyl, C1-5 alkyl, amino, hydroxide, or halogen; wherein when L1is amino-C1-5 alkylene or an amide, L1is bonded via nitrogen to both R1and R1”, wherein one or both ofDocket No. 084284.00356R1or R1can be hydrogen, when L2is amino-Ci-5 alkylene or amide, L2is bonded via nitrogen to both R2and R2, wherein one or both of R2or R2can be hydrogen, when L3is amino-Ci-5 alkylene or amide, L3is bonded via nitrogen to both R3and R3, wherein one or both of R3or R3can be hydrogen, when L1is ester, ether, or an oxygen-Ci-5 alkylene, only R1is bonded via oxygen to L1and R1is void, when L2is ester, ether, or an oxygen-Ci-5 alkylene, only R2is bonded via oxygen to L2and R2is void, and when L3is ester, ether, or an oxygen-Ci-5 alkylene, only R3is bonded via oxygen to L3and R3is void. In some embodiments, when the dotted line is void, the group it is attached to is also void. In some embodiments, the dotted lines is a void or a single bond.

[0115] In this configuration, A is connected to L1at ring position 2 of A, A is connected to V of Formula (I) at ring position 5 of A, B is connected to V at ring position 2 of B, and B is connected to L2at ring position 5 of B. In some embodiments, when E is connected to L3at ring position 2, E is connected to V at ring position 5 and when E is connected to L3at ring position 5, E is connected to V at ring position 2. In some embodiments, at least one of L1, L2, or L3is amino-Ci-5 alkylene.

[0116] In certain embodiments, a subset of compounds of Formula (I) and Formula (IH) includes those of Formula (li):E / C-i-4z\R3’r3(li)Docket No. 084284.00356

[0117] wherein A, B, and E are independently a heteroaryl, aryl, cycloalkyl, heterocycloalkyl, a furan or tetrahydrofuran, wherein A, B, and E are optionally and independently substituted at any of the carbon positions with a linear or branched Ci-is alkyl, Ci-is alkoxyl-alkylene, hydrogen, halo-Ci-5 alkyl, C1-5 alkyl, amino, hydroxide, or halogen; V from Formula (I) is Ci alkylene-D, and D is E-L3(R3)(R3”); L1, L2, and L3from Formula (I) and Formula (IH) are each amino-Ci-5 alkylene optionally substituted with one or more of C1-5 alkyl, amino, or hydroxide; R1, R1, R2, R2, R3, and R3are independently hydrogen, a linear or branched Ci-36 alkyl, optionally inserted with one or more ester, carbonate (-O-(C=O)-O-), amino (-NH2), amide, or oxygen (-O-), and optionally substituted with one or more of halo-Ci-5 alkyl, C1-5 alkyl, amino, hydroxide, or halogen.

[0118] In some embodiments, V from Formula (I) may independently be from any of the following groups:

[0119] wherein a, b, c, d, and e are independently selected from 1, 2, 3, 4, and 5. In some embodiments, C1-5 alkylene may be optionally substituted with one or more of halo-Ci-5 alkyl, Ci-5 alkyl, amino, hydroxide, or halogen, C3-6 cycloalkyl ring, along any carbon position on C1-5 alkylene chain. In some embodiments, alkyl groups -(CH2)a-, -(CH2)b-, -(CH2)c, -(CH2)d-, -(CH2)e-may optionally be substituted at any carbon position with one or more of hydrogen, halo-Ci-5 alkyl, C1-5 alkyl, amino, hydroxide, or halogen.Docket No. 084284.00356

[0120] In some embodiments, L1, L2, and L3may be independently selected from any of the following groups wherein W is A or B and Y is halogen:

[0121] wherein j and k are independently selected from 1, 2, 3, 4, and 5 wherein the sum of j and k is 2, 3, 4, or 5. In some embodiments, alkyl groups -(CH2)j- or -(CH2)k- may optionallyDocket No. 084284.00356be substituted at any one of carbon positions with one or more of hydrogen, halo-Ci-5 alkyl, amino, hydroxide, or halogen.

[0122] In some embodiments, tail groups, i.e., R1, R1, R2, R2”, R3, and R3may independently comprise hydrogen, a linear or branched Ci-36 alkyl optionally inserted with one or more of any one of the subgroups including an ester, carbonate (-O-(C=O)-O-), amino (-NH2), amide, oxygen (-O-), an acetal linker linkage (-O-CH2-O-). In some embodiments, tail groups, i.e., R1, R1, R2, R2, R3, and R3may further be substituted along any one or more carbon atoms along the Ci-36 alkyl with one or more of hydrogen, halo-Ci-5 alkyl, amino, hydroxide, or halogen.

[0123] In some embodiments, R1, R1, R2, R2”, R3, and R3may be independently selected from any of the following groups, wherein Z is L1, L2or L3, and Y is halogen:Docket No. 084284.00356

[0124] wherein f is selected from any value from 1 to 36, i is selected from 1, 2, 3, 4, and 5, and g and h are independently selected from any value from 1 to 36, wherein the sum of g and h is from any value from 2 to 36. In some embodiments, Ci-36 alkyl may be optionally inserted with one or more of any one of the subgroups including an ester, carbonate (-O-(C=O)-O-), amino (-NH2), amide, oxygen (-O-), an acetal linker linkage (-O-CH2-O-). In some embodiments, Ci-36 alkyl may be optionally substituted at any carbon position with one or more of hydrogen, halo-Ci-5 alkyl, C1-5 alkyl, amino, hydroxide, or halogen, along any carbon position on Ci-36 alkyl chain.

[0125] In some embodiments, alkyl groups -(CH2)f-, -(CH2)g- or -(CH2)h- may optionally be inserted with one or more of any one of the subgroups including an ester, carbonate (-O-(C=O)-O-), amino (-NH2), amide, oxygen (-O-), an acetal linker linkage (-O-CH2-O-) to provide a combination of subgroups on the alkyl chain. In some embodiments, alkyl groups -(CH2)i-, -(CH2)f-, -(CH2)g- or -(CH2)h- may optionally be substituted at any carbon position with one or more of hydrogen, halo-Ci-s alkyl, C1-5 alkyl, amino, hydroxide, or halogen.

[0126] In certain embodiments, a subset of compounds of Formula (I) includes those of Formula (IJ):Docket No. 084284.00356

[0127] L1and L2are independently ester, ether, amide, amino-Ci-5 alkylene, or oxygen-Ci-5 alkylene, wherein amino-Ci-5 alkylene and oxygen-Ci-5 alkylene may be optionally substituted with one or more of C1-5 alkyl, amino, and hydroxide, and wherein amino-Ci-5 alkylene or oxygen-C1-5 alkylene are attached to A or B at the alkylene end; R1, R1”, R2and R2are independently hydrogen, a void (represented by dotted line) or a linear or branched Ci-36 alkyl, optionally inserted with one or more ester, carbonate (-O-(C=O)-O-), amino (-NH2), amide, or oxygen (-O-), and optionally substituted with one or more of halo-Ci-5 alkyl, C1-5 alkyl, amino, hydroxide, or halogen; wherein F1and F2are independently selected from the group consisting of a linear or branched Ci 18 alkyl, Ci-isalkoxyl-alkylene, hydrogen, halo-Ci-5 alkyl, C1-5 alkyl, C1-6 cycloalkyl-Ci-is alkylene, amino, hydroxide, or halogen. In some embodiments, when the dotted line is void, the group it is attached to is also void. In some embodiments, the dotted line is a void or a single bond.

[0128] In certain embodiments, a subset of compounds of Formula (I) includes those of Formula (IK):

[0129] L1and L2are independently ester, ether, amide, amino-Ci-5 alkylene, or oxygen-Ci-5 alkylene, wherein amino-Ci-5 alkylene and oxygen-Ci-s alkylene may be optionally substituted with one or more of C1-5 alkyl, amino, and hydroxide, and wherein amino-Ci-s alkylene or oxygen-C1-5 alkylene are attached to A or B at the alkylene end; R1, R1”, R2and R2are independently hydrogen, a void (represented by dotted line) or a linear or branched Ci-36 alkyl, optionally inserted with one or more ester, carbonate (-O-(C=O)-O-), amino (-NH2), amide, or oxygen (-O-), and optionally substituted with one or more of halo-Ci-5 alkyl, C1-5 alkyl, amino, hydroxide, or halogen;Docket No. 084284.00356wherein F1, F2, F3, and F4are independently selected from the group consisting of a linear or branched Ci-18 alkyl, Ci-isalkoxyl-alkylene, hydrogen, halo-Ci-5 alkyl, C1-5 alkyl, C1-6 cycloalkyl-C1-18 alkylene, amino, hydroxide, or halogen. In some embodiments, when the dotted line is void, the group it is attached to is also void. In some embodiments, the dotted line is a void or a single bond.

[0130] In certain embodiments, a subset of compounds of Formula (I) includes those of Formula (IL):

[0131] L1, L2, and L3are independently ester, ether, amide, amino-Ci-5 alkylene, or oxygen-Ci-5 alkylene, wherein amino-Ci-5 alkylene and oxygen-Ci-5 alkylene may be optionally substituted with one or more of C1-5 alkyl, amino, and hydroxide, and wherein amino-Ci-5 alkylene or oxygen-Ci-5 alkylene are attached to A or B at the alkylene end; R1, R1”, R2, R2”, R3, and R3are independently hydrogen, a void (represented by dotted line) or a linear or branched Ci-36 alkyl, optionally inserted with one or more ester, carbonate (-O-(C=O)-O-), amino (-NH2), amide, or oxygen (-O-), and optionally substituted with one or more of halo-Ci-5 alkyl, C1-5 alkyl, amino, hydroxide, or halogen; wherein F1, F2, F3, F4, F5, and F6are independently selected from the group consisting of a linear or branched Ci-18 alkyl, Ci-isalkoxyl-alkylene, hydrogen, halo-Ci-5 alkyl, Ci-5 alkyl, C1-6 cycloalkyl-Ci-18 alkylene, amino, hydroxide, or halogen. In some embodiments, when the dotted line is void, the group it is attached to is also void. In some embodiments, the dotted line is a void or a single bond.Docket No. 084284.00356

[0132] In some embodiments, a subset of compounds of Formula (I) includes the following structures:

[0133] In some embodiments, a subset of compounds of Formula (I) includes the following structures Fl to Fll, wherein each of the furan or tetrahydrofuran may be optionally and independently substituted at any of the carbon position with a linear or branched Ci-18 alkyl, Ci-isDocket No. 084284.00356alkoxyl-alkylene, hydrogen, halo-Ci-5 alkyl, C1-5 alkyl, C1-6 cycloalkyl-Ci-is alkylene, amino,the following groups,^^C1-36OH OH^^^ _ OH Y00 Z}i° \Docket No. 084284.003560ZAACA OH NH, / VO^OVYOSA / XA 0 / )i O 0 OH OH [ O I 1. XA O Y YYO OHT000\wherein Y is halogen, f is selected from any value from 1 to 36, i is selected from 1, 2, 3, 4, and 5, and g and h are independently selected from any value from 1 to 36, wherein the sum of g and h is from any value from 2 to 36. In some embodiments, Ci-36 alkyl may be optionally inserted with one or more of any one of the subgroups including an ester, carbonate (-O-(C=O)-O-), amino (-NH2), amide, oxygen (-O-), an acetal linker linkage (-O-CH2-O-). In some embodiments, Ci-36 alkyl may be optionally substituted at any carbon position with one or more > oof hydrogen, halo-Ci-5 alkyl, C1-5 alkyl, amino, hydroxide, or halogen, along any carbon position 0on Ci -36 alkyl chain.

[0135] In some embodiments, alkyl groups -(CH2)f-, -(CH2)g- or -(CH2)h- may optionally be inserted with one or more of any one of subgroups including an ester, carbonate (-O-(C=O)-O-), amino (-NH2), amide, oxygen (-O-), an acetal linker linkage (-O-CH2-O-) to provide a combination of subgroups on alkyl chain. In some embodiments, alkyl groups -(CH2)i-, -(CH2)f-, -(CH2)g- or -(CH2)h- may optionally be substituted at any carbon position with one or more of hydrogen, halo-Ci-5 alkyl, C1-5 alkyl, amino, hydroxide, or halogen.

[0136] wherein R1, R1, R2, R2, R3, and R3are independently selected from the following structures T1-T6 below:Docket No. 084284.00356

[0137] In some embodiments, R1, R1, R2, R2”, R3, and R3are independently selectedDocket No. 084284.00356

[0138] In some embodiments, the compound of Formula (I) is selected from the following compounds:OH F1T2— ooF1T3F1T4F1T6Docket No. 084284.00356F2T1F2T2F2T3F2T4F2T6Docket No. 084284.00356F3T1F3T2F3T3Docket No. 084284.00356F3T4F3T5F3T6Docket No. 084284.00356F4T1F4T2F4T3Docket No. 084284.00356F4T4F4T5Docket No. 084284.00356F4T6F5T1F5T2Docket No. 084284.00356Docket No. 084284.00356F5T5F5T6F6T1Docket No. 084284.00356F6T2F6T3F6T4Docket No. 084284.00356F6T5F6T6F7T1Docket No. 084284.00356F7T2F7T3Docket No. 084284.00356Docket No. 084284.00356F7T6F8T1F8T2Docket No. 084284.00356Docket No. 084284.00356F8T5F8T6F9T1Docket No. 084284.00356F4T4F4T5F4T6F5T1Docket No. 084284.00356F5T2F5T3F5T4F5T5Docket No. 084284.00356F5T6F6T1F6T2Docket No. 084284.00356F6T3F6T4F6T5Docket No. 084284.00356F6T6F7T1Docket No. 084284.00356Docket No. 084284.00356F7T2F7T3Docket No. 084284.00356F7T4F7T5Docket No. 084284.00356Docket No. 084284.00356Docket No. 084284.00356Docket No. 084284.00356Docket No. 084284.00356

[0139] Tn some embodiments, the compound of Formula (T) is selected from the following compounds:A B V LI L2 Rl’ Rl” R2’ R2”1. o^X Bond Bond A^°XVoid / xfto^oA2 Bond Bond AA>XVoid3. O^X Bond Bond 0 Voidz-\^0'Z^'0'^h'4 Bond Bond O Void A'% / z^oA / ^5 Bond Bond 0 O ^0^ A^0^c / V> AOA< 6. O^X Bond Bond 0 o A^o^o^A*FOA *< *h * *h 7}S< X Bondx^io-^oAA^N£8 Bond^01^ AZX^O'^S'OX^7 A^o^o^ Afto^o^Docket No. 084284.003569, p^ Bond / ^0^0^'\JI Z^O^cA Z^O^cA10..o^X. 0ZAJ AXN AXN A^icAo^i AAT A^oAr 11. O^X 0 or^0^0^vv / ^> AftcA / (% >^Ar AAT 12^0^ <7r> xA / > A A^(ACA xA^ 13 j z°-^A..o^X 0> o <Nfto^oA7 AX / ^ry AAO4 M-AA 'Wi'IA 14^0^ <7 AX AXNzA'ZX0'<^z^^O'ZX‘Oz^‘ AAA ZA(A(A 15..o~^X J p-^- / ~\3 A-N£ / ^N£z^io / s-oAz^io'^-o'<^Z^AO4z>ftoAA 16 0 o ^0^ ^0^ / V'y AAo4 T^o-U 17. o^X / VyZ> ACAOAZAA(AZ$AO4 zX^Ao^frDocket No. 084284.0035618, / NA 0 019z^fV^< A20z.#^rAZ-^^O / Xoz^‘z-\^o^^o^721..o^X J z°'A 0 0x^io^o^22 0 0ry x^O^O^ x^io^o^b23x\(io^oA^01^ >^^PYzjftAo424. O^X. o^X ZAAAz-\^O^XOx^h'^^Y

[0140] wherein f, g, and h are independently between 1-36 and the sum of g and h is between 2-36, and wherein one or more carbon atoms on -(CH2)f- -(CH2)g- or -(CH2)h- are optionally substituted with one or more of halo-Ci-5 alkyl, C1-5 alkyl, amino, hydroxide, or halogen.Docket No. 084284.00356Lipid Nanoparticles (LNPs)

[0141] In another aspect, the present disclosure also features a lipid nanoparticle composition comprising a lipid component comprising a compound according to Formula (I), (IA), (IB), (IC), (ID), (IE), (IF), (IG), (IH) or (li) as described herein.

[0142] As used herein, a “lipid component” is that component of a nanoparticle composition that includes one or more lipids. For example, the lipid component may comprise an ionizable lipid comprising a compound according to Formula (I), (IA), (IB), (IC), (ID), (IE), (IF), (IG), (IH) or (li) and a helper lipid component including but not limited to one or more PEGylated-lipid, structural-lipid, or phospholipids.

[0143] In some embodiments, the lipid nanoparticle formulation may comprise a furanderived ionizable lipid and at least one helper lipid. As used herein, the term “Helper lipid” refers to a lipid component of a lipid nanoparticle formulation that is distinct from the cationic or ionizable lipid and that functions to stabilize the lipid nanoparticle structure and / or facilitate intracellular delivery of an associated therapeutic agent, including by modulating membrane properties, promoting lipid phase transitions, or enhancing endosomal escape, without being primarily responsible for electrostatic complexation of the therapeutic agent. In some embodiments, helper lipid is a neutral or zwitterionic lipid incorporated into a lipid nanoparticle to support particle assembly, structural integrity, and biological performance, including by influencing lipid packing, curvature, fluidity, or membrane fusion behavior. In certain embodiments, the helper lipid contributes to destabilization of the endosomal membrane following cellular uptake, thereby enhancing release of the therapeutic agent into the cytosol.

[0144] In some embodiments, the helper lipid may be a phospholipid, a structural lipid, or a pegylated (i.e., PEG-modified)-lipid. As used herein, the term “PEG lipid” or “pegylated lipid”Docket No. 084284.00356refers to a lipid comprising a polyethylene glycol component. As used herein, the term “phospholipid” is a lipid that includes, a phosphate moiety and one or more carbon chains, e.g., unsaturated fatty acid chains. The phospholipid may comprise one or more multiple (e.g. double or triple bond) bonds (e.g. one or more unsaturated bonds). Certain phospholipids may promote fusion with the membrane. For example, a cationic phospholipid may interact with one or more negatively charged phospholipids of a membrane (e.g., a cell membrane or an intracellular membrane). The fusion of the phospholipid to the membrane may allow one or more elements of the lipid-containing composition to pass through the membrane, thereby allowing, for example, the delivery of one or more elements to the cell.

[0145] In some embodiments, the phospholipid comprises Dilauroyl lecithin (DLPC), Dimyristoyl phosphatidylcholine (DMPC), Dioleoyl lecithin (DOPC), Dipalmitoyl phosphatidylcholine (DPPC), Distearoyl phosphatidylcholine (DSPC), Dioleoyl phosphatidylcholine (DUPC), Palmitoyl oleoyl phosphatidylcholine (POPC), 1,2-Di-O-octadecyl-sn-glycero-3 -phosphocholine, 1 -Oleoyl -2-cholesteryldimethylsuccinoyl-sn-glycero-3-phosphocholine, 1 -Hexadecyl-sn-glycero-3 -phosphocholine, 1,2-Divinyl-sn-glycero-3 -phosphocholine, l,2-Diarylacyl-sn-glycero-3-phosphocholine, 1,2-Dioleoyl-sn-glycero-3phosphorylethanolamine (DOPE), 1, 2-Di-phytanoyl-sn-glycero-3 -phosphoethanolamine, 1,2-Distearoyl-sn-glycero-3-phosphoethanolamine, l,2-Diethenol-sn-glycero-3-phosphoethanolamine, l,2-Divinyl-sn-glycero-3 -phosphoethanolamine, l,2-Diaryl-sn-glycero-3-phosphoethanolamine, l,2-Dithiohexaenoate-sn-glycero-3-phosphoethanolamine, 1,2-dioleoyl-sn-glycero-3-phosphate-(l -glycerol) sodium salt (DOPG), and combinations thereof.Docket No. 084284.00356

[0146] In some embodiments, the structural lipid comprises cholesterol, coprosterol, fecosterol, sitosterol, ergosterol, campesterol, stigmasterol, brassicasterol, tomatidine, tomatine, ursolic acid, alpha-tocopherol, and combinations thereof.

[0147] In some embodiments, the pegylated-lipid comprises 1,2-dimyristoyl-sn-glycerol methoxypolyethylene glycol (DMG-PEG), l,2-distearoyl-sn-glycero-3-phosphoethanolamine-N-[amino(polyethylene glycol)] (DSPE-PEG), PEG-disteryl glycerol (DSG-PEG), PEG-dipalmetoleyl, PEG-dioleyl, PEG-distearyl, PEG-diacylglycamide (DAG-PEG), PEG-dipalmitoyl phosphatidylethanolamine (DPPE-PEG), or PEG-1, 2-dimyristyloxlpropyl-3-amine (DMA-c-PEG), and combinations thereof. In some embodiments, the lipid moiety of the PEG lipids includes those having lengths from about C14 to about C22. In some embodiments, a PEG moiety has a size of about 1000, 2000, 5000, 10,000, 15,000 or 20,000 daltons, i.e., PEG1000, PEG2000, PEG5000, PEG10000, PEG15000, or PEG20000 respectively.

[0148] In some embodiments, the molar ratio between the furan-derived ionizable lipid and the helper lipid component (i.e., phospholipid + structural lipid + pegylated lipid) is from about 20 / 200 to about 20 / 40, or about 20 / 180, about 20 / 160, about 20 / 140, about 20 / 120, about 20 / 100, about 20 / 90, about 20 / 80, about 20 / 70, about 20 / 60, about 20 / 50, about 20 / 40, or between any two aforementioned values of furan-derived ionizable lipid / helper lipid component.

[0149] In some embodiments, the molar ratio between the furan-derived ionizable lipid and phospholipid is from about 20 / 80 to about 20 / 10, about 20 / 70, about 20 / 60, about 20 / 50, about 20 / 40, about 20 / 30, about 20 / 20, about 20 / 10, or between any two aforementioned values of furanderived ionizable lipid / phospholipid.

[0150] In some embodiments, the molar ratio between the furan-derived ionizable lipid and structural lipid is from about 20 / 80 to about 20 / 10, about 20 / 70, about 20 / 60, about 20 / 50,Docket No. 084284.00356about 20 / 40, about 20 / 30, about 20 / 20, about 20 / 10, or between any two aforementioned values of furan-derived ionizable lipid / structural lipid.

[0151] In some embodiments, the molar ratio between the furan-derived ionizable lipid and pegylated lipid is from about 20 / 2 to about 20 / 0.5, about 20 / 1.75, about 20 / 1.5, about 20 / 1.25, about 20 / 1, about 20 / 0.75, about 20 / 0.5, about 20 / 0.25, or between any two aforementioned values of furan-derived ionizable lipid / pegylated lipid.

[0152] In some embodiments, the molar ratio between the furan-derived ionizable lipid and each component of the helper lipid i.e., phospholipid, structural lipid, and pegylated lipid (furan-derived ionizable lipid / phospholipid / structural lipid / pegylated lipid) is from about 20 / 80 / 80 / 2 to about 20 / 10 / 10 / 0.5 of furan-derived ionizable lipid / pegylated lipid, or any values between therein (e.g., any values encompassed by the following ratio: 1-20 / 5-80 / 5-80 / 0.1-2). In some embodiments the molar ratio between the furan-derived ionizable lipid and each component of the helper lipid i.e., phospholipid, structural lipid, and pegylated lipid (furan-derived ionizable lipid / phospholipid / structural lipid / pegylated lipid) is from about 20 / 30 / 40 / 0.75 of furan-derived ionizable lipid / pegylated lipid.

[0153] In some embodiments, the disclosed LNP further comprises a therapeutic agent. In certain embodiments, the therapeutic agent is encapsulated by the LNP. In some embodiments, the therapeutic agent comprises carbohydrates, protein, lipids, nucleic acids, or combinations thereof. As used herein, the term “nucleic acid” refers to a molecule of two or more nucleotides or alternative nucleotides. The term, “nucleotide” refers to a nucleoside including a phosphate group. The term “nucleoside” refers to a compound containing a sugar molecule (e.g., a pentose or ribose) or derivative thereof in combination with an organic base (e.g., a purine or pyrimidine) or a derivative thereof (also referred to herein as a “nucleobase”). Examples of nucleic acids includeDocket No. 084284.00356but are not limited to DNA, RNA, tRNA (transfer RNA), mRNA (messenger RNA), siRNA (small interfering RNA), miRNA (micro-RNA), shRNA (short hairpin RNA), ncRNA (non-coding RNA), aptamers, ribozymes, and shorter oligonucleotide sequences of any of the foregoing. Alterations of the base, sugar, and phosphate moiety of a nucleotide are encompassed by this definition.

[0154] In some embodiments, the therapeutic agent is mRNA. An mRNA may encode any polypeptide of interest, including any naturally or non-naturally occurring or otherwise modified polypeptide. A polypeptide encoded by an mRNA may be of any size and may have any secondary structure or activity. In some embodiments, a polypeptide encoded by an mRNA may have a therapeutic effect when expressed in a cell. In some embodiments, mRNA comprises an mRNA encoding for firefly luciferase (FLuc), green fluorescent protein (GFP), glial cell line-derived neurotrophic factor (GDNF), ere recombinase (Cre), brain-derived neurotrophic factor (BDNF), nerve growth factor (NGF), neurturin (NRTN), insulin-like growth factor- 1 (IGF-1), vascular endothelial growth factor (VEGF), aducanumab, donanemab, gantenerumab, solanezumab, crenezumab, etanercept, trastuzumab, natalizumab, briakinumab, bevacizumab, or lecanemab. In some embodiments, the therapeutic agent may be delivered to prevent, ameliorate, or treat a disease or other medical condition associated with neurodegenerative diseases, infectious diseases, genetic disorders, stem cell disorders, and metabolic diseases, including but not limited to Parkinson’s disease, Amyotrophic lateral sclerosis (ALS), peripheral neuropathies, spinal cord injury, Huntington’s disease, and ischemic neural injury.

[0155] In certain embodiments, the LNP comprises a mass ratio of furan-derived ionizable lipid to therapeutic agent ranging from about 5:1 to about 30:1, about 10: 1, about 15:1, about 20: 1, about 25:1, about 30:1, or between any two aforementioned values. All or a portion of theDocket No. 084284.00356therapeutic agent nucleic acid may be encapsulated in the lipid nanoparticles. In some embodiments, the LNP may exhibit encapsulation efficiency of at least about 90% (e.g., at least about 85%, at least about 80%, at least about 75%, at least about 70%, at least about 65%, at least about 60%, at least about 55%, at least about 50%).

[0156] In some embodiments, the LNP may have a mean hydrodynamic diameter ranging from about 50 nm to about 1000 nm (e.g., from about 50 nm to about 100 nm, about 100 nm to about 200 nm, about 200 nm to about 300 nm, about 300 nm to about 400 nm, about 400 nm to about 500 nm, about 500 nm to about 600 nm, about 600 nm to about 700 nm, about 700 nm to about 800 nm, about 800 nm to about 900 nm, about 900 nm to about 1000 nm, or about 50 nm, about 100 nm, about 150 nm, about 200 nm, about 250 nm, about 300 nm, about 350 nm, about 400 nm, about 450 nm, about 500 nm, about 550 nm, about 600 nm, about 650 nm, about 700 nm, about 750 nm, about 800 nm, about 850 nm, about 900 nm, about 950 nm, or about 1000 nm). As used herein, the term “hydrodynamic diameter” refers to a measurement of a particle's size that takes into account the particle's movement through a liquid. “Hydrodynamic diameter” represents the size of a sphere that diffuses at the same rate as the particle under observation, taking into account any solvation layer or dynamic interaction with the solvent.

[0157] In some embodiments, the LNP may have a mean poly dispersity index (PDI) from about 0.1 to about 0.8 (e.g., about 0.1, about 0.2, about 0.3, about 0.4, about 0.5, about 0.6, about 0.7, about 0.8 and any range between aforementioned values). As used herein, the term “poly dispersity index” refers to a measurement of heterogeneity of particle sizes. If particles in the population are uniform, the resulting size distribution will be narrow and the PDI small, determining that the sample is monodisperse. For heterogeneous particle populations, the size distribution will be broader and the PDI increases, determining that the sample is polydisperse.Docket No. 084284.00356

[0158] In some embodiments, the disclosed LNP may be used for intracellular delivery of a therapeutic agent. In some embodiments, the delivery of the LNP may comprise contacting a cell or administering to a subject a lipid nanoparticle comprising a therapeutically effective amount of a therapeutic agent. As used herein, the term “subject” encompasses any animal, but preferably a mammal, e g., a human, non-human primate, a dog, a cat, or a mouse. More preferably, the subject is a human. As used herein, the term “delivery” refers to providing an entity to a target site. For example, delivering a therapeutic and / or prophylactic agent to a subject may include administering a nanoparticle composition containing a therapeutic and / or prophylactic agent to a subject (e.g., by intravenous, intramuscular, intradermal, intrathecal, or subcutaneous routes). Administering a nanoparticle composition to a mammal or mammalian cell may involve contacting one or more cells with the nanoparticle composition.

[0159] In certain embodiments, the lipid nanoparticle (LNP) comprises a furan-derived ionizable lipid, phospholipid, structural lipid, and pegylated-lipid as disclosed above. In some embodiments, the furan-derived ionizable lipid has a structure of formula (I) disclosed above. In some embodiments, the furan-derived ionizable lipid has a structure of formula (II) disclosed above. In some embodiments, the phospholipid is DOPE. In some embodiments, the structural lipid is cholesterol. In some embodiments, the pegylated-lipid is DMG-PEG2000. In some embodiments, the therapeutic agent is an mRNA encoding for firefly luciferase (FLuc).

[0160] In some embodiments, a pharmaceutical composition comprising the present LNP is disclosed herein, including a therapeutically effective amount of at least one therapeutic agent and / or one or more pharmaceutically acceptable excipient, carrier, or diluent. As used herein, the term “pharmaceutically acceptable excipient, carrier, or diluent” includes compounds, materials, compositions, and / or dosage forms that are, within the scope of sound medical judgment, suitableDocket No. 084284.00356for use in contact with the tissues of human beings and animals without excessive toxicity, irritation, allergic response, or other problem or complication, commensurate with a reasonable benefit / risk ratio. In some embodiments, the pharmaceutical compositions may be in a form suitable for parenteral administration. Depending upon the therapeutic application, for in vivo administration, the disclosed LNP herein may be administered to a subject in need thereof intravenously, intradermally, intramuscularly, intrathecally, or subcutaneously.

[0161] In some embodiments, the pharmaceutical compositions may be in the form of a sterile injectable aqueous or suspension, which may be formulated according to known procedures. A sterile injectable preparation may also be a sterile injectable suspension in a non-toxic parenterally-acceptable buffer. In other embodiments, the pharmaceutical composition may be lyophilized resulting in the form of a dry powder, wherein the dry powder can be later reconstituted for administration as needed. Dry powder compositions may further comprise bulking agents, for example, sucrose or trehalose.

[0162] Further provided herein is a pharmaceutical kit comprising a pharmaceutical composition comprising the disclosed LNP having a therapeutically effective amount of at least one therapeutic agent. Such kits may further comprise various conventional pharmaceutical kit components such as containers comprising pharmaceutically acceptable adjuvants, diluents, or carriers, and additional containers readily apparent to those skilled in the art. Instructions, either as inserts or as labels, indicating quantities of the components to be administered, guidelines for administration, and / or guidelines for mixing the components, can also be included in the kit.

[0163] It will be appreciated by persons skilled in the art that embodiments described herein are not limited to what has been particularly shown and described. Rather, the scope of the invention is defined by the claims which follow. It should further be understood that the aboveDocket No. 084284.00356description is only representative of illustrative examples of embodiments. The description has not attempted to exhaustively enumerate all possible variations. The alternate embodiments may not have been presented for a specific component of a nanocarrier, or a step of the method, and may result from a different combination of described constituents, or other un-described alternate embodiments may be available for a composition, kit or method, is not to be considered a disclaimer of those alternate embodiments. It will be appreciated that many of those un-described embodiments are within the literal scope of the following claims, and others are equivalent.EXAMPLES EXAMPLE 1: Chemical synthesis of tetrahydrofuran-derived ionizable lipids

[0164] In the reaction schemes described herein, multiple stereoisomers may be produced. When no particular stereoisomer is indicated, it is understood to mean all possible stereoisomers that could be produced from the reaction. A person of ordinary skill in the art will recognize that the reactions can be optimized to give one isomer preferentially, or new schemes may be devised to produce a single isomer. If mixtures are produced, techniques such as preparative thin layer chromatography, preparative HPLC, preparative chiral HPLC, or preparative SFC may be used to separate the isomers.

[0165] Compounds of the present disclosure can be prepared in a variety of ways using commercially available starting materials, compounds known in the literature, or from readily prepared intermediates, by employing standard synthetic methods and procedures either known to those skilled in the art, or which will be apparent to the skilled artisan in light of the teachings herein. Standard synthetic methods and procedures for the preparation of organic molecules and functional group transformations and manipulations can be obtained from the relevant scientific literature or from standard textbooks in the field. Although not limited to any one or severalDocket No. 084284.00356sources, classic texts such as Smith, M. B., March, J., Marchs Advanced Organic Chemistry. Reactions, Mechanisms, and Structure, 5thedition, John Wiley & Sons: New York, 2001; Greene, T. W., Wuts, P. G. M., Protective Groups in Organic Synthesis, 3rdedition, John Wiley & Sons: New York, 1999; R. Larock, Comprehensive Organic Transformations, VCH Publishers (1989); L. Fieser and M. Fieser, Fieser andFieser's Reagents for Organic Synthesis, John Wiley and Sons (1994); and L. Paquette, ed., Encyclopedia of Reagents for Organic Synthesis, John Wiley and Sons (1995), incorporated by reference herein, are useful and recognized reference textbooks of organic synthesis known to those in the art. The following descriptions of synthetic methods are designed to illustrate, but not to limit, general procedures for the preparation of compounds of the present disclosure.

[0166] Furan is a five-membered aromatic heterocycle with four carbon atoms and one oxygen atom, commonly used as a precursor in the synthesis of various pharmaceutical compounds. Tetrahydrofuran (THF) is the fully saturated version of furan, widely employed in pharmaceutical formulations for solubilizing drugs. Mono-, di-, and tri-furan cores were conjugated with various lipid chains. Similar reactions were conducted for mono- and ditetrahydrofuran-derived ionizable lipids. A representative synthetic route to tetrahydrofuranderived ionizable lipids is outlined in the following reaction mechanism.Docket No. 084284.00356OTMSB'''' i l. AD-mix β, MeSO2NH2HO OH NIS, DCM DCM / t-BuOH c

[0167] A reaction between trans-1,4-dibromobutene A and allyl magnesium bromide B was initiated to yield a triene, which underwent Sharpless asymmetric dihydroxylation to give diol C. Upon treatment with N-iodosuccinimide (NIS), cyclization of diol C gave compound E.Subsequently, compound E was subjected to azidation to get F, followed by reduction with LiAlH4, leading to the formation of diamine G. The hydrophobic tails were then installed to the core amine via epoxide ring-opening reactions or reductive amination reactions, affording the corresponding furan-derived ionizable lipids. 1H nuclear magnetic resonance (1H NMR) spectra confirmed the chemical structures of the lipids.

[0168] Synthesis of compound C. A solution of trans-1,4-dibromobutene A (6.68 g, 31.25 mmol, 1.0 equiv) in Et2O (15 mL) was added dropwise to a solution of allyl magnesium bromide B (1.0 M in Et2O, 100 mL, 100 mmol, 3.2 equiv) at 0°C. The resulting mixture was stirred overnight, allowing the temperature to increase to room temperature (25°C). The reaction was then quenched by the addition of aqueous acetic acid (AcOH) (10.5 M, 3.46 mL, 36.33 mmol, 2.18 g AcOH, 2.08 mL) at 0°C. The mixture was poured onto ice (ca. 156 g). The two layers were separated, and the aqueous one was extracted with diethyl ether (Et2O) (3 x 50 mL). The combinedDocket No. 084284.00356organic layers were washed with aqueous NaHCO3 (10% w / w, 2 x 50 mL) and brine, dried over MgSO4, fdtered and the solvent was removed in vacuo.

[0169] The crude product was purified by filtration through a short silica plug (elution with pentane) to afford the triene product (2.7 g, 88.6%) as a colorless oil. Following a reported procedure, triene product (1.0 eq., 19.8 mmol, 2.70 g) was added to a suspension of AD-mix (15.4 g) and methanesulfonamide (1.05 g, 11.1 mmol, 1.01 eq.) in a solution of tert-butanol and water (100 mL / 100 mL). The resulting mixture was stirred at 0°C for 7 h and then the reaction was quenched by the addition of Na2SO3 (ca. 15 g) and allowed to warm to room temperature. The aqueous layer was extracted with ethyl acetate (EtOAc) and the combined organic layers were washed with brine, dried over MgSO4, filtered and the solvent was removed in vacuo, after purification by column chromatography diol C was obtained as colorless oil (1.0 g, 29.7%).

[0170] Synthesis of compound E. Under a nitrogen atmosphere, compound C was added to a solution of D in dichloromethane (DCM) at -78 °C. The reaction mixture was stirred for 10 min at -78 °C and N-Iodosuccinimide (NIS) was added to the mixture at -78 °C and stirred for 2 h. After being stirred for 12 h at room temperature, the reaction was quenched with sat. Na2S2O3 aqueous solution. And the mixture was extracted with DCM. The organic layer was dried over Na2SO4 and evaporated in vacuo. The residue was purified by silica gel column chromatography to give compound E as a colorless oil. 1H NMR (400 MHz, Chloroform-d) δ4.13 - 4.00 (m, 2H), 3.97 - 3.84 (m, 2H), 3.33 - 3.25 (m, 2H), 3.23 - 3.11 (m, 2H), 2.14 - 2.01 (m, 2H), 1.96 - 1.85 (m, 2H), 1.85 - 1.68 (m, 4H).

[0171] Synthesis of azide F. Sodium azide (993 mg, 15.28 mmol) was added to a solution of E (1.61 g, 3.82 mmol) inDMF (35 mL). After being stirred for 4 h at 70 °C, the reaction mixture was quenched with water. The mixture was extracted with Et2O, and then the combined organicDocket No. 084284.00356was washed with water. The organic layer was dried over anhydrous Na SO4 and evaporated under reduced pressure. The residue was purified via silica gel chromatography to give the azide compound F (690 mg, 71.6%) as a colorless oil.1H NMR (300 MHz, Chloroform-d) 84.22 - 4.09 (m, 2H), 3.97 - 3.83 (m, 2H), 3.41 (dd, J= 12.8, 4.4 Hz, 2H), 3.30 (dd, J= 12.8, 5.6 Hz, 2H), 2.09 - 1.88 (m, 4H), 1.86 - 1.68 (m, 4H).

[0172] Synthesis of diamine G. A suspension of LiAlH4 (8.19 mmol, 311 mg) in 25 mL of dry tetrahydrofuran (THF) was stirred while a solution of azide compound F (0.69 g, 2.73 mmol) in dry THF was added dropwise at 0 °C over a period of 10 min. The reaction mixture was warmed to room temperature and stirred for 3 h then quenched by the sequential addition of water (0.4 mL), a 15 % solution of NaOH (0.4 mL), and water (1.2 mL), the mixture was stirred for 1 h. The white precipitate was filtered off and washed with ether or THF (20 mL). The filtrate was dried over Na2SO4, filtered, and concentrated in vacuo to give diamine G as a colorless oil.EXAMPLE 2: General synthetic procedure for the epoxide ring-opening reaction between diamines and epoxide.

[0173] To a solution of diamine (0.2 mmol) in 2 mL of dry (ethanol) EtOH, epoxide (1.0 mmol) was added. Then the mixture was warmed to 90 °C and kept stirring for 12 h. Thin Layer Chromatography showed total consumption of diamine, EtOH was removed under reduced pressure. The residue was purified via silica gel chromatography (0%-100% [mixture of 3% NH4OH, 22% MeOH (methanol) in DCM] in DCM) to give desire products.EXAMPLE 3: General synthetic procedure for reductive amination of diaminesDocket No. 084284.003566 F7T1

[0174] To a solution of diamine G (0.2 mmol) in THF (4 mL), aldehyde (1.0 mmol) was added, the mixture was kept stirring for 30 min at room temperature. Then NaBH(OAc)3 (254 mg, 1.2 mmol) was added to the above solution, and the resulting mixture was stirred for 12 h. Aq. NaHCO3 solution (15 mL) was added to quench the reaction. The aqueous solution was extracted with DCM (15 mL x 3 times), the organic phase was combined and dried over anhydrous Na2SC>4, fdtered, and concentrated under reduced pressure. The residue was purified via silica gel chromatography (0%-100% [mixture of 3% NH4OH, 22% MeOH in DCM] in DCM) to give the desired compounds. The synthetic procedure of F6T1 is similar to Fl 1T6.EXAMPLE 4: Preparation and characterization of LNPs.

[0175] LNPs were prepared using an established four-component formulation and were used to encapsulate mRNA agents: Synthesized furan-derived ionizable lipid, DOPE, cholesterol, and DMG-PEG2k. The lipid nanoparticles were formulated with DOPE, cholesterol, and DMG-PEG2k at a molar ratio of lipid / DOPE / cholesterol / DMG-PEG2k = 20 / 30 / 40 / 0.75. The initial mass ratio of furan-derived ionizable lipids to mRNA was 10:1. A control SMI 02 lipid was formulated with DSPC, cholesterol, and DMG-PEG2k at a molar ratio of SM102 / DSPC / cholesterol / DMG-PEG2k = 50 / 10 / 38.5 / 1.5, with an SM102 lipid-to-mRNA mass ratio of 11.03:1. Hydrodynamic diameter and poly dispersity index (PDI) of LNPs were measured using a NanoZS Zetasizer (Malvern), which also facilitated zeta potential measurements. LNP morphology was visualized by Cryo-TEM (Thermo Scientific Glacios), as previously described. mRNA encapsulation efficiency was measured by a modified Quant-iT RiboGreen RNA assay (Invitrogen). For in vivoDocket No. 084284.00356batch experiments and subsequent administration, furan-derived ionizable lipids were individually formulated with DOPE, cholesterol, and DMG-PEG2k at a molar ratio of lipid / DOPE / cholesterol / DMG-PEG2k = 20 / 30 / 40 / 0.75 and the mass ratio of furan-derived ionizable lipids to mRNA was 10:1.

[0176] The hydrodynamic diameter of all lipid nanoparticles comprising different furanderived ionizable lipids was measured using Zetasizer NanoZS (Malvern, UK) and determined to be 170 nm and ranged from 50 nm to 1,000 nm. The average poly dispersity index of all was measured using Zetasizer NanoZS (Malvern, UK) and determined to be 0.13 and ranged from 0.01 to 0.8. The average Zeta potential of the lipid nanoparticles was measured using Zetasizer NanoZS (Malvern, UK) and determined to be -0.25 mV and ranged from -12 mV to 5 mV. The average encapsulation efficiency of the lipid nanoparticles was measured using Quant-iT™ RiboGreen RNA Assay and determined to be 92 % and ranged from 45% to 95%.EXAMPLE 5: Batch-based experiments for in vivo evaluation.

[0177] The performance of furan-derived ionizable lipids for mRNA delivery into the brain through meningeal lymphatic vessels (MLVs), a special circulatory network between deep cervical lymph nodes (dCLNs) and the brain was evaluated. To assess the delivery efficacy of this lymphatic route to the brain, LNPs were administered subcutaneously near the dCLNs in the mouse neck. A batch-based approach was used to reduce the number of animals, time, and cost. Briefly, the LNPs were formulated to encapsulate mRNA encoding firefly luciferase (FLuc) and categorized into 11 groups based on head group structures. In each batch, prior to the subcutaneous injection, 6 LNPs with the same head group but different tails were mixed.

[0178] The mRNA delivery efficiency was appraised in vivo. The mixed LNPs were subcutaneously administered into the neck of mice, followed by In Vivo Imaging System (IVIS)Docket No. 084284.00356after 6 hours. The luminescent intensity as seen in FTG. 1 shows LNPs containing F2 group LNPs containing hydroxyl-modified mono-furan lipids exhibited superior mRNA delivery efficiency compared to Fl batch LNPs with methyl ester modifications. Nevertheless, LNPs formulated with di-furan-derived ionizable lipids did not outperform their mono-furan counterparts. Similarly, LNPs formulated with tri-furan-derived ionizable lipids did not improve the mRNA delivery performance. Interestingly, tetrahydrofuran lipids with four-tail conjugation in the F10 group demonstrated markedly higher mRNA delivery efficacy compared to their furan analogs. Additionally, replacing the hydroxyl group on tetrahydrofuran lipids with two additional lipid tails significantly enhanced mRNA delivery efficiency, as shown in LNP batches F9 and F10. Even though increasing the number of tetrahydrofuran cores to di-tetrahydrofuran resulted in a decrease in mRNA delivery efficacy, lipids in the Fl 1 group still maintained performance within the top three among various furan and tetrahydrofuran-based lipid formulations, superior to di-furan derived lipids. Therefore, the top three head groups, F2, F10, and Fl 1 were identified for further study. Particularly, F10 group LNPs displayed the most exceptional delivery efficiency.

[0179] Next, the effects of lipid tail structures on mRNA delivery were also assessed. Diverse functional groups were incorporated in the tail chains, such as ester, carbonate, and acetal. Following a similar approach to the head structure assessment, all LNPs were classified into 6 batches based on their tail structures, each batch comprising 11 LNPs. A mixture of 11 LNPs was injected into mice as described above, and then mRNA delivery efficiency was determined. As shown in FIG. 2, LNPs comprising tails with ester and acetal ether linkage were the most effective in mRNA delivery, which was over 3.3-fold higher than other LNPs as measured by luminescence intensity. Hence, two top-performing tails, T3 and T6, were identified for further characterization.Docket No. 084284.00356These orthogonal batch-based tests provided a quick method to select the head groups and lipid tails for further study.

[0180] Based on the above data, exemplary LNPs containing ionizable lipids with high-performing head groups (F2, F10, and Fl 1) and tails (T3, T6) were selected for further evaluation. SM102 LNPs, an FDA-approved LNP, was included as a control. In light of the remarkable delivery performance of the F 10 head group, furan-derived ionizable lipids containing F10 and each different tail group (T1-T6) were also tested in mice. As seen in luminescence intensity data in FIG. 3, results show that F6T1 and Fl 1T6 LNPs performed higher mRNA delivery efficiency to the brain than other LNPs. As outlined in Table 1, according to the predicted LogD (as determined through MarvinSketch), the LogD value of F2T6 lipid was 5.43, whereas the LogD values of other lipids were from 9.08 to 18.21, showing 6.9 to 42.85-fold higher luminescence intensity than F2T6 LNP in the brain. In particular, both F6T1 and Fl 1T6 lipids were near 11.0, whose luminescence intensities were up to ~40-fold higher (F6T1) than that of F2T6 lipid, indicating the LogD may be an important factor for LNP delivery to the brain via MLVs.Table 1. Predicted LogD values of different lipids.Lipid F2T3 F2T6 F5T3 F5T4 F5T5 F5T6 F6T1 F6T2 F6T5 F7T1 LogD 9.08 5.43 15.15 12.55 17.73 11.83 11.33 10.41 18.21 10.89 Luminescence6 90 12 23 19 27 16 68 16 50 42 85n36 19 n3961intensity*The luminescence intensity of F2T6 LNP for in vivo brain delivery of FLuc mRNA was defined as 1

[0181] As seen in FIG. 4, even though F6T1 LNPs showed similar luminescence intensity with Fl 1T6 LNPs in the brain, their distribution in other major organs such as the liver, spleen, and lung are lower than F7T1 LNPs. Subsequently, F6T1 and F7T1 LNPs were selected for additional delivery studies.EXAMPLE 6: Cellular trafficking of LNPsDocket No. 084284.00356

[0182] To study the cellular trafficking of these new LNPs, a series of experiments in Neuro-2a (N2a) cells, a mouse neuroblast cell line was performed. Mostly consistent with the results in mice, LNPs formulated with lipids containing F2, F10, and Fll head groups showed high mRNA delivery efficiency. Specifically, F6T1 and Fl 1T6 LNPs exhibited superior mRNA delivery to SMI 02 LNP. It was identified that F2T3 LNPs displayed high mRNA delivery efficiency at the cellular level but low efficiency for in vivo mRNA delivery, indicating the differences between in vitro and in vivo experiments. Afterwards, the cellular uptake of LNPs in N2a cells were evaluated. These LNPs were formulated with ionizable lipids containing different head groups (Fl-Fl 1) and Alexa-Fluor 647-labeled mRNA while maintaining the same tail structure (T6).

[0183] As shown in FIGS. 5A and 5B, F1T6 and F1T5 LNPs demonstrated the highest cellular uptake efficiency. Additionally, LNPs formulated with F2T6, F5T2, F2T5 and F5T1 also exhibited outstanding cellular internalization. Notably, these high-performing lipids all featured a two-tail structure, suggesting these two-tail structures were beneficial to cellular uptake. However, despite F7T1 and F6T2 LNPs achieving relatively low cellular uptake, they exhibited high expression of FLuc mRNA, implying that factors beyond internalization, such as enhanced endosomal escape capability likely accounted for the high mRNA delivery efficiency of F6T2 and Fl 1T6 LNP. the enhanced endosomal escape capability likely accounted for the high mRNA delivery efficiency of F6T2 and Fl 1T6 LNPs, despite their lower cellular uptake.

[0184] Additionally, cells incubated with Calcein alone exhibited punctuated fluorescence, indicating the endosomal sequestration of the internalized dye. In contrast, diffused green fluorescence was observed throughout the cytoplasm in cells treated withF6T2 and Fl 1T6 LNPs. The wide distribution of diffuse green fluorescence was not detected in cells treated with the otherDocket No. 084284.00356LNPs. The widespread fluorescence suggested LNP-mediated endosomal membrane escape, enabling the cytoplasmic release of mRNA. Additionally, Pearson’s correlation analysis revealed reduced colocalization of green fluorescence with the red signal of LNPs in cells treated with F6T2 and F7T1 LNPs, reconfirming the endosomal escape. Thus, the enhanced endosomal escape capability likely accounted for the high mRNA delivery efficiency of the F6T2 and Fl 1T6 LNPs.

[0185] Based on the delivery data, the mechanism of LNPs bypassing BBB via MLVs route in vivo was explored. The lymphatic delivery of Alexa-Fluor 647-labeled mRNA-loaded LNPs to the MLVs was investigated following their subcutaneous injection into the dCLNs at the mouse neck. The results indicated colocalization of LNPs and MLVs. This colocalization suggested a lymphatic pathway for LNP transport from the dCLNs to the meninges, potentially explaining their ability to bypass the BBB. Moreover, the GFP mRNA was also transported by the lymphatic delivery pathway. The GFP expression in the meninges and MLVs were monitored showing that LNPs could not only reach the meninges via lymphatic routes but, also deliver mRNA to brain cells via the MLVs.

[0186] To further examine mRNA delivery in different brain cell types, F6T1 and Fl 1T6 LNPs were loaded with GFP mRNA, and GFP expression was quantified by flow cytometry. As shown in FIG. 6A, F6T1 and F7T1 LNPs preferentially delivered mRNA to neurons. Particularly, Fl 1T6 LNP demonstrated superior neuron delivery efficiency (16.4% GFP+) and the neuron delivery efficiency of F6T1 LNP was 8.8% GFP+. Moreover, GFP mRNA was also delivered to microglia and astrocytes by F6T1 and F7T1 LNPs. In contrast, SM102 LNPs showed minimal GFP mRNA delivery efficiency to brain cells, not only in neurons (1.28% GFP+) but also in microglia (1.16% GFP+) and astrocytes (1.79% GFP+). Furthermore, as seen in FIG.Docket No. 084284.003566B, a small fraction of brain capillary endothelial cells (BCECs) also exhibited GFP expression when treated with F6T1, Fl 1T6, and SM102 LNPs.

[0187] To evaluate the potential toxicity of furan-derived LNPs, blood urea nitrogen (BUN), alanine aminotransferase (ALT), and aspartate aminotransferase (AST) levels were measured in mice treated with various LNP formulations. As seen in FIGS. 7A-7C, all these markers remained within normal ranges, indicating no adverse effects on hepatic or renal function following administration of F6T1 and Fl 1T6 LNPs at the tested dose.

[0188] The potential of LNPs for functional mRNA delivery in an Ail4 mouse model was also assessed. The Ail4 mouse line is genetically engineered with a LoxP-flanked stop cassette to suppress tdTomato expression at baseline but enables its induction upon exposure to Cre recombinase. In this experiment, Cre recombinase mRNA-loaded F6T1 and F7T1 LNPs were subcutaneously administered to the neck of Ail4 mice. In mice treated with PBS, tdTomato expression was negligible 7 days post-administration. As seen in FIGS. 8A-8C, following the delivery of Cre mRNA using F6T1 and Fl 1T6 LNPs, tdTomato signals were detected throughout most regions of the brain, including neurons, microglia, and astrocytes. In contrast, administration of SM102 LNPs resulted in a low tdTomato signal. Specifically, the tdTomato expression in brain cells was quantified at 8.93% and 13.0% in neurons, 7.0% and 12.1% in microglia, and 9.9% and 6.5% in astrocytes for F6T1 and Fl 1T6 LNPs. Notably, the tdTomato signals induced by F6T1 and Fl 1T6 LNPs were significantly higher than those elicited by SM102 LNPs. Collectively, these results highlighted the potential of F6T1 and Fl 1T6 LNPs as promising carriers to deliver various mRNA to important brain cell types via MLVs routes, especially neurons, which facilitated the expression of functional proteins in brain cells.Docket No. 084284.00356

[0189] The foregoing merely illustrates the principles of the disclosure. Any examples set forth in this specification are not intended to be limiting and merely set forth some of the many possible embodiments for the appended claims. Those skilled in the art will readily recognize various modifications and changes that may be made without following the example embodiments and applications illustrated and described herein, and without departing from the true spirit and scope of the following claims.

[0190] All references cited and / or discussed in this specification are incorporated herein by reference in their entirety and to the same extent as if each reference was individually incorporated by reference.

Claims

Docket No. 084284.00356What is claimed is:

1. A lipid compound comprising a compound of Formula (I):, R2"R1'A B^R2' (I),wherein A is a furan or tetrahydrofuran;B is a bond, furan or tetrahydrofuran;L1and L2are independently ester (-O-(C=O)- or -(C=O)-O-), ether, amide (-(C=O)-N- or -N-(C=O)-), amino-Ci-5 alkylene, or oxygen-Ci-5 alkylene, wherein amino-Ci-5 alkylene and oxygen-Ci-5 alkylene may be optionally substituted with one or more selected from the group consisting of C1-5 alkyl, amino, and hydroxide, and wherein amino-Ci-5 alkylene, or oxygen-Ci-5 alkylene are attached to A or B at alkylene end;R1, R1, R2and R2are independently hydrogen, void or a linear or branched Ci-36 alkyl, optionally inserted with one or more ester, carbonate (-O-(C=O)-O-), amino (-NH2), amide, or oxygen (-O-), and optionally substituted with one or more selected from the group consisting of halo-Ci-5 alkyl, C1-5 alkyl, amino, hydroxide, and halogen, wherein the void is represented by a dotted line, wherein when R1or R2is not void, it is connected via a single bond;V is a bond, a linear or branched C1-5 alkylene optionally substituted with one or more selected from the group consisting of C1-5 alkyl, (-[C(L4)(L5)]-), C3-6 cycloalkyl, and C1-5 alkylene-D, wherein D is E-L3(R3)(R3);L4and I? are independently C1-5 alkyl or a hydrogen;E is a furan or tetrahydrofuran;L3is an ester, ether, amide, amino-Ci-s alkylene, or oxygen-Ci-5 alkylene, wherein amino-C1-5 alkylene and oxygen-Ci-5 alkylene may optionally be substituted with one or more of C1-5 alkyl, amino, and hydroxide, and wherein amino-Ci-5 alkylene, or oxygen-Ci-5 alkylene are attached to E at alkylene end;R3andR3are independently hydrogen, void or a linear or branched Ci-36 alkyl, optionally inserted with one or more selected from the group consisting of ester, carbonate, amino (-NEE), amide, and oxygen (-O-), and optionally substituted with one or more selected from the group consisting of halo-Ci-5 alkyl, C1-5 alkyl, amino, hydroxide, and halogen;Docket No. 084284.00356wherein when L1is amino-Ci-5 alkylene or an amide, L1is bonded via nitrogen to both R1and R1”, wherein one or both of R1or R1is hydrogen,when L1is ester, ether, or an oxygen-Ci-5 alkylene, only R1is bonded via oxygen to L1and R1is void,when L2is amino-Ci-5 alkylene or amide, L2is bonded via nitrogen to both R2and R2”, wherein one or both of R2or R2is hydrogen,when L2is ester, ether, or an oxygen-Ci-5 alkylene, only R2is bonded via oxygen to L2and R2is void,when L3is amino-Ci-5 alkylene or amide, L3is bonded via nitrogen to both R3and R3, wherein one or both of R3or R3is hydrogen, andwhen L3is ester, ether, or an oxygen-Ci-5 alkylene, only R3is bonded via oxygen to L3and R3is void.

2. The lipid compound of claim 1, when B is a furan or a tetrahydrofuran,A is connected to L1at ring position 2 and connected to V at ring position 5 of A;1 L1^2 / °x5^V / / 3 / \4F1F2A, and.B is connected to L2at ring position 5 and connected to V at ring position 2 of B;1 V — 2 / °x§^L2\\ vF1F2B wherein F1and F2are independently selected from the group consisting of a linear or branched Ci-is alkyl, Ci-isalkoxyl-alkylene, hydrogen, halo-Ci-5 alkyl, C1-5 alkyl, Ci-6 cycloalkyl-Ci-i8 alkylene, amino, hydroxide, and halogen.Docket No. 084284.003563. The lipid compound of claim 1 or claim 2, whereinV is -[C(L4)(L5)]-;L4and L5are independently Ci s alkyl or a hydrogen;L1and L2are independently amino-Ci-5 alkylene optionally substituted with one or more selected from the group consisting of alkyl, amino, and hydroxide; andR1, R1, R2, and R2are independently hydrogen, a linear or branched Ci-36 alkyl, optionally inserted with one or more selected from the group consisting of ester, carbonate, amino, amide, and oxygen, and optionally substituted with one or more selected from the group consisting of halo-Ci-5 alkyl, C1-5 alkyl, amino, hydroxide, and halogen.

4. The lipid compound of any one of claims 1-3, whereinV is a bond;L1and L2are independently amino-Ci-5 alkylene optionally substituted with one or more selected from the group consisting of alkyl, amino, and hydroxide; andR1, R1”, R2and R2are independently hydrogen, a linear or branched Ci-36 alkyl, optionally inserted with one or more selected from the group consisting of ester, carbonate, amino, amide, and oxygen, and optionally substituted with one or more selected from the group consisting of halo-Ci-5 alkyl, C1-5 alkyl, amino, hydroxide, and halogen,wherein A is connected to B at ring position 5 of A and ring position 2 of B, andA is connected to L1at ring position 2 of A, and B is connected to L2at ring position 5 of B,1 1 L1^2 / °X 5 2 / OSSL2' / V 'I3^-44 3^-44F1F2F3F4A Bwherein F1, F2, F3, and F4are independently selected from the group consisting of a linear or branched Ci-is alkyl, Ci-18 alkoxyl-alkylene, hydrogen, halo-Ci-5 alkyl, C1-5 alkyl, C1-6 cycloalkyl-Ci-18 alkylene, amino, hydroxide, and halogen.Docket No. 084284.003565. The lipid compound of any one of claims 1-4, wherein the compound of Formula (I) is selected from:Docket No. 084284.003566. The lipid compound of claim 5, wherein R1, R1, R2, R2, R3, or R3are independently selected from:OH0* 'h0, wherein f, g, and h are independently between 1-36 and sum of g and h is between 2-36,one or more carbon atom on –(CH2)f–, –(CH2)g– or –(CH2)h– is optionally substituted with one or more selected from the group consisting of halo-Ci-5 alkyl, C1-5 alkyl, amino, hydroxide, and halogen.Docket No. 084284.003567. The lipid compound of any one of claims 1-6, comprising a compound of Formula (TA):R1X" R2 / L" A'“ ^R2'R1(IA),wherein V and B are each bond to form a single bond between A and L2,A is a furan or tetrahydrofuran;L1and L2are independently ester, ether, amide, amino-Ci-5 alkylene, or oxygen-Ci-5 alkylene, wherein amino-Ci-5 alkylene and oxygen-Ci-5 alkylene may be optionally substituted with one or more selected from the group consisting of C1-5 alkyl, amino, and hydroxide, and wherein amino-Ci-5 alkylene, or oxygen-Ci-5 alkylene are attached to A at alkylene end;R1, R1”, R2and R2are independently hydrogen, a void or a linear or branched Ci-36 alkyl, optionally inserted with one or more selected from the group consisting of ester, carbonate, amino, amide, and oxygen, and optionally substituted with one or more selected from the group consisting of halo-Ci-5 alkyl, C1-5 alkyl, amino, hydroxide, and halogen,wherein when L1is amino-Ci-5 alkylene or amide, L1is bonded via nitrogen to both R1and R1”, wherein one or both of R1or R1is hydrogen,when L1is ester, ether, or an oxygen-Ci-5 alkylene, only R1is bonded via oxygen to L1and R1is void,when L2is amino-Ci-5 alkylene or amide, L2is bonded via nitrogen to both R2and R2”, wherein one or both of R2or R2is hydrogen,when L2is ester, ether, or an oxygen-Ci-5 alkylene, only R2is bonded via oxygen to L2and R2is void,A is connected to L1at ring position 2 of A, andA is connected to L2at ring position 5 of A,Docket No. 084284.00356wherein F1and F2are independently selected from the group consisting of a linear or branched Ci-is alkyl, Ci-isalkoxyl-alkylene, hydrogen, halo-Ci-5 alkyl, C1-5 alkyl, C1-6 cycloalkyl-C1-18 alkylene, amino, hydroxide, and halogen.

8. The lipid compound of claim 7, wherein,L1is amino-Ci-5 alkylene optionally substituted with one or more selected from the group consisting of C1-5 alkyl, amino, and hydroxide;L2is oxygen-Ci-5 alkylene optionally substituted with one or more selected from the group consisting of C1-5 alkyl, amino, and hydroxide; andR1, R1”, R2, and R2are independently hydrogen, a linear or branched Ci-36 alkyl, optionally inserted with one or more selected from the group consisting of ester, carbonate, amino, amide, and oxygen, and optionally substituted with one or more selected from the group consisting of halo-Ci-5 alkyl, C1-5 alkyl, amino, hydroxide, and halogen.

9. The lipid compound of claim 7 or claim 8, whereinL1and L2are independently amino-Ci-5 alkylene optionally substituted with one or more selected from the group consisting of alkyl, amino, and hydroxide; andR1, R1, R2, and R2are independently hydrogen, a linear or branched Ci-36 alkyl, optionally inserted with one or more selected from the group consisting of ester, carbonate, amino, amide, and oxygen, and optionally substituted with one or more selected from the group consisting of halo-Ci-5 alkyl, C1-5 alkyl, amino, hydroxide, and halogen.

10. The lipid compound of any one of claims 1-9, comprising a compound of Formula (IH):wherein A, B and E are independently a furan or tetrahydrofuran;L1, L2, and L3are independently ester, ether, amide, amino-Ci-5 alkylene, or oxygen-Ci-5 alkylene, wherein amino-Ci-5 alkylene and oxygen-Ci-5 alkylene may optionally be substitutedDocket No. 084284.00356with one or more selected from the group consisting of C1-5 alkyl, amino, and hydroxide, and wherein amino-Ci-5 alkylene, or oxygen-Ci-5 alkylene are attached to A, B, and E at alkylene end;R1, R1, R2, R2, R3and R3are independently hydrogen, a void or linear or branched Ci-36 alkyl, optionally inserted with one or more selected from the group consisting of ester, carbonate, amino, amide, and oxygen, and optionally substituted with one or more selected from the group consisting of halo-Ci-5 alkyl, C1-5 alkyl, amino, hydroxide, and halogen, wherein the void is represented by a dotted line, wherein when R1’, R2, or R3is not void, it is connected via a single bond,wherein when L1is amino-Ci-s alkylene or an amide, L1is bonded via nitrogen to both R1and R1”, wherein one or both of R1or R1is hydrogen,when L2is amino-Ci-5 alkylene or amide, L2is bonded via nitrogen to both R2and R2”, wherein one or both of R2or R2is hydrogen,when L3is amino-Ci-5 alkylene or amide, L3is bonded via nitrogen to both R3and R3”, wherein one or both of R3or R3is hydrogen,when L1is ester, ether, or an oxygen-Ci-5 alkylene, only R1is bonded via oxygen to L1and R1is void,when L2is ester, ether, or an oxygen-Ci-5 alkylene, only R2is bonded via oxygen to L2and R2is void, andwhen L3is ester, ether, or an oxygen-Ci-5 alkylene, only R3is bonded via oxygen to L3and R3is void.

11. The lipid compound of claim 10, whereinL1, L2, and L3are independently an amino-Ci-5 alkylene optionally substituted with one or more selected from the group consisting of C1-5 alkyl, hydrogen, amino, and hydroxide; and R1, R1, R2, R2, R3, and R3are independently hydrogen, a void or a linear or branched Ci-36 alkyl, optionally inserted with one or more selected from the group consisting of ester, carbonate, amino, amide, and oxygen, and optionally substituted with one or more selected from the group consisting of halo-Ci-5 alkyl, C1-5 alkyl, amino, hydroxide, and halogen.

12. A lipid nanoparticle comprising the lipid compound of any one of claims 1-11 and a helper lipid, wherein the helper lipid is selected from the group consisting of phospholipid, a structuralDocket No. 084284.00356lipid, pegylated-lipid, and a combination thereof, and the lipid nanoparticle encloses a therapeutic agent.

13. The lipid nanoparticle of claim 12, wherein the phospholipid is selected from the group consisting of Dilauroyl lecithin (DLPC), Dimyristoyl phosphatidylcholine (DMPC), Dioleoyl lecithin (DOPC), Dipalmitoyl phosphatidylcholine (DPPC), Di stearoyl phosphatidylcholine (DSPC), Dioleoyl phosphatidylcholine (DUPC), Palmitoyl oleoyl phosphatidylcholine (POPC), 1.2-Di-O-octadecyl-sn-glycero-3 -phosphocholine, l-Oleoyl-2-cholesteryldimethylsuccinoyl-sn-glycero-3-phosphocholine, 1 -Hexadecyl -sn-glycero-3 -phosphocholine, l,2-Divinyl-sn-glycero-3-phosphocholine, l,2-Diarylacyl-sn-glycero-3-phosphocholine, 1,2-Dioleoyl-sn-glycero-3phosphorylethanolamine (DOPE), 1, 2-Di-phytanoyl-sn-glycero-3 -phosphoethanolamine, 1,2-Distearoyl-sn-glycero-3-phosphoethanolamine, l,2-Diethenol-sn-glycero-3-phosphoethanolamine, l,2-Divinyl-sn-glycero-3 -phosphoethanolamine, l,2-Diaryl-sn-glycero-3-phosphoethanolamine, l,2-Dithiohexaenoate-sn-glycero-3-phosphoethanolamine, 1,2-dioleoyl-sn-glycero-3-phosphate-(l -glycerol) sodium salt (DOPG), and combinations thereof.

14. The lipid nanoparticle of claim 12 or claim 13, wherein the structural lipid is selected from the group consisting of cholesterol, coprosterol, fecosterol, sitosterol, ergosterol, campesterol, stigmasterol, brassicasterol, tomatidine, tomatine, ursolic acid, alpha-tocopherol, and combinations thereof.

15. The lipid nanoparticle of any one of claims 12-14, wherein the pegylated-lipid is selected from the group consisting of 1,2-dimyristoyl-sn-glycerol methoxypolyethylene glycol (DMG-PEG), 1.2-distearoyl-sn-glycero-3-phosphoethanolamine-N-[amino(polyethylene glycol)] (DSPE-PEG), PEG-disteryl glycerol (DSG-PEG), PEG-dipalmetoleyl, PEG-dioleyl, PEG-distearyl, PEG-diacylglycamide (DAG-PEG), PEG-dipalmitoyl phosphatidylethanolamine (DPPE-PEG), or PEG-l,2-dimyristyloxlpropyl-3-amine (DMA-c-PEG), and combinations thereof.

16. The lipid nanoparticle of any one of claims 12-15, wherein the phospholipid is DOPE, the structural lipid is cholesterol, and the pegylated-lipid is DMG-PEG,Docket No. 084284.00356wherein a molar ratio of the furan-derived lipid, the DOPE, the cholesterol, and the DMG-PEG is about 20 / 30 / 40 / 0.75.

17. The lipid nanoparticle of claim 16, further comprising an mRNA encapsulated in the lipid nanoparticle, wherein a mass ratio of the furan-derived lipid to the mRNA is from about 5:1 to 30:1.

18. The lipid nanoparticle of claim 17, wherein the mRNA encodes for firefly luciferase (FLuc), green fluorescent protein (GFP), glial cell line-derived neurotrophic factor (GDNF), ere recombinase (Cre), brain-derived neurotrophic factor (BDNF), nerve growth factor (NGF), neurturin (NRTN), insulin-like growth factor-1 (IGF-1), vascular endothelial growth factor (VEGF), aducanumab, donanemab, gantenerumab, solanezumab, crenezumab, etanercept, trastuzumab, natalizumab, briakinumab, bevacizumab, or lecanemab.

19. A method of intracellular delivery of a therapeutic agent, comprising, contacting a cell or administering to a subject, a lipid nanoparticle of any one of claims 12-18.

20. A composition comprising the lipid nanoparticles of any one of claims 12-18.

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