Nanoparticle formulations for the delivery of DNA to adipose tissue

Abstract

The present invention generally relates at least in part to gene delivery vehicles, and more specifically, in some embodiments, DNA delivery vehicles for the treatment of human or veterinary conditions.

Description

[0001] NANOPARTICLE FORMULATIONS FOR THE DELIVERY OF DNA TO ADIPOSE TISSUE RELATED APPLICATIONS

[0002] This application claims the benefit of priority under 35 U. S . C . § 119 (e) of U. S . Provisional Application Serial No . 63 / 712 , 158 , filed on October 25, 2024 , the entire contents of which are incorporated herein by reference .

[0003] FIELD OF THE INVENTION

[0004] The present invention generally relates at least in part to gene delivery vehicles, and more specifically, in some embodiments, DNA delivery vehicles for the treatment of human or veterinary conditions .

[0005] SUMMARY OF THE INVENTION

[0006] Protein therapies, whether administered subcutaneously or intravenously, are limited by sawtoothing pharmacokinetics, which result in undesirable side effects at the pharmacokinetic peak and insufficient efficacy at the pharmacokinetic trough for some subcutaneously administered therapies . These effects are highly pronounced in some therapies, for example GLP-1 recepor agonists for weight loss or the treatment of type 2 diabetes . In said therapies the side effects cause high rates of discontinuation of treatment, by some estimates as high as 68% after 1 year . In contrast, gene therapies provide a more homogeneous protein production profile at steady state, however, the dose cannot be adjusted following initial treatments . It has been found that the most optimal delivery site for gene therapies are subcutaneous adipose tissues, as they are a non-life sustaining organ and not required for function, unlike for example the liver . These features enable dose adjustment, make the treatment safe, and allow for the utilization of adipocytes as a drug production center following delivery of genetic material that encodes a therapeutic transgene . Provided herein are optimized formulations for the delivery of DNA to adipocytes and preadipocytes . The formulations enable the replacement of protein treatment regimens with single injection gene therapies that can be optionally adjusted or removed without significant adverse effects to the organism and serve as a means of optimizing protein delivery pharmacokinetics thereby improving safety and efficacy.

[0007] Effective delivery of therapeutic biologies for treatment of disease can depend on the ability of the therapy to reach the target tissue or cells, enter the cells, and then escape from the internal (endosomal) trafficking system of the cells to provide therapeutic benefit . An effective treatment is therefore inherently limited by the stability of the biologically active compound in blood or cells, as well as its ability to traverse cellular membranes . Delivery of nucleic acids is especially problematic, due to the presence of nucleases in extracellular fluids and prevention of entrance into cells by their negative charge . Gene therapy, which aims to solve this, often entails the use of viruses (e . g . , lentiviruses or adenoassociated viruses [AAV] ) . While the use of these viruses solves stability problems and issues with moving across cellular membranes, viruses can cause local or systemic inflammatory responses, and immune memory prevents therapies based on viruses from being re-dosed if needed. Furthermore, patients may already have neutralizing antibodies against specific viral vector, limiting the efficacy of any therapeutic using that vector . As an example, an estimated 50% of people have neutralizing antibodies against AAV after the age of 3. Lipid nanoparticles (LNPs) are capable of stabilizing and transporting genetic cargo into cells without overt immune memory responses . Provided herein are optimized formulations of LNPs for delivery of genetic cargo (DNA) to adipocytes of the subcutaneous tissues . The LNP formulations for the delivery of DNA-based therapeutic transgenes to adipose tissues can be for safe and effective treatment of disease .

[0008] In an aspect is a nanoparticle formulation for the delivery of genetic cargo to subcutaneous adipose tissues, which comprises :

[0009] a . at least one ionizable lipid, or lipidoid, or lipid-like molecule, or intracellular vesicle-interacting peptide, and b . a structural element comprised of at least one polymer or structural and cationic lipid, and

[0010] c . optionally at least one cholesterol or cholesterol analog or cholesterol substitute, and

[0011] d. optionally a targeting element, and

[0012] e . a genetic cargo of 1. 5-7. 0kb in length encoding at least one therapeutic gene and formulated in an N to P ratio of 3.5 to 9.5.

[0013] In an embodiment of any one of the compositions or methods provided herein, the ionizable lipid is a cationic lipid. In an embodiment of any one of the compositions or methods provided herein, the ionizable lipid is a dimethylammonium-propane or analog. In an embodiment of any one of the compositions or methods provided herein, the ionizable lipid is 18 : 0 DAP ( 1 , 2-distearoyl-3-dimethylammonium-propane) or analog . In an embodiment of any one of the compositions or methods provided herein, the ionizable lipid is 16 : 0 DAP ( 1, 2-dipalmitoyl-3-dimethylammonium-propane) or analog. In an embodiment of any one of the compositions or methods provided herein, the ionizable lipid is 14 : 0 DAP ( 1, 2-dimyristoyl-3-dimethylammonium-propane) or analog . In an embodiment of any one of the compositions or methods provided herein, the ionizable lipid is 18 : 1 DAP or DODAP ( 1, 2-dioleoyl-3-dimethylammonium-propane) or analog. In an embodiment of any one of the compositions or methods provided herein, the ionizable lipid is Lipid C24 (bis (2-octyldodecyl) 3 , 3 ' - ( ( 4- ( 4-methylpiperazin-l-yl) butyl) azanediyl) dipropionate) or analog . In an embodiment of any one of the compositions or methods provided herein, the ionizable lipid is ATX-001 ( 8, 8 '- [ [2- [ [2- ( dimethylamino) ethyl] thio] acetyl] imino] bis-octanoic acid, 1, 1 '-di- (2Z) -2-nonen-l-yl ester) or analog . In an embodiment of any one of the compositions or methods provided herein, the ionizable lipid is Lipid A9 (bis (2-butyloctyl) 10- (N- (3- (dimethylamino) propyl) nonanamido) nonadecanedioate) or analog. In an embodiment of any one of the compositions or methods provided herein, the ionizable lipid is CIN-16645 ( 9Z, 12Z-octadecadienoic acid, 3- [4 , 4- bis (octyloxy) -1-oxobutoxy] -2- [ [ [ [3 —

[0014] (diethylamino) propoxy] carbonyl] oxy] methyl] propyl ester) or analog. In an embodiment of any one of the compositions or methods provided herein, the ionizable lipid is Lipid 23 (8— ( (2— ( (4—

[0015] (dimethylamino) butanoyl) oxy) ethyl) (10-oxo-10- (tridecan-7-yloxy) decyl) amino) octyl 2-hexyldecanoate) or analog. In an embodiment of any one of the compositions or methods provided herein, the ionizable lipid is Lipid 29 (8- [ [8- [ ( 1-ethylnonyl) oxy] -8-oxooctyl] [3- [ [2- (methylamino) -3, 4-dioxo-l-cyclobuten-l-yl] amino] propyl] amino] -octanoic acid, 1-octylnonyl ester) or analog . In an embodiment of any one of the compositions or methods provided herein, the ionizable lipid is Lipid 8 ( 4- (dimethylamino) -butanoic acid, 2- [di- ( 9Z, 12Z) -9, 12-octadecadien-l-ylamino] ethyl ester) or analog . In an embodiment of any one of the compositions or methods provided herein, the ionizable lipid is 1014 (15, 19, 28-tris (2-hydroxytetradecyl) -15, 19, 24 , 28-tetraazadotetracontane-13, 30-diol) or analog. In an embodiment of any one of the compositions or methods provided herein, the ionizable lipid is 246C10 ( 1 , 1 ' , 1 ' ' , 1 ' ' ' - [ 1 , 4-piperazinediylbis (3, 1-propanediylnitrilo) ] tetrakis-2-dodecanol) or analog. In an embodiment of any one of the compositions or methods provided herein, the ionizable lipid is 98N12-5 (Nl, N16-didodecyl-4 , 7 , 13 -tris [3- (dodecyl amino) -3 -oxopropyl ] -4, 7, 10, 13-tetraazahexadecanediamide) or analog . In an embodiment of any one of the compositions or methods provided herein, the ionizable lipid is YK-009 ( 6- [ [ 4- (decyloxy) -4-oxobutyl] (2-hydroxyethyl) amino] -hexanoic acid, 2-octyldecyl ester) or analog. In an embodiment of any one of the compositions or methods provided herein, the ionizable lipid is CL4F8-6 (7- (4- (dipropylamino) butyl) -7 -hydroxytride cane- 1, 13 -diyl bis (2-hexyloctanoate) ) or analog . In an embodiment of any one of the compositions or methods provided herein, the ionizable lipid is IC8 ( 1, 1 ' , 1 ' ' , 1 ' ' ' - ( (piperazine-1, 4-diylbis (propane-3, 1-diyl) ) bis (azanetriyl) ) tetrakis (tetradecan-2-ol) ) or analog. In an embodiment of any one of the compositions or methods provided herein, the ionizable lipid is Lipid III-45 (2-butyl-octanoic acid, l, l '- [ [ (3-hydroxypropyl) imino] di-9, 1-nonanediyl] ester) or analog . In an embodiment of any one of the compositions or methods provided herein, the ionizable lipid is ATX-100 ( 4, 4 '- [ [ [ [3- ( dimethylamino) propyl] thio] carbonyl] imino] bis-butanoic acid, 1, 1 ' -bis ( 1-heptyloctyl) ester) or analog. In an embodiment of any one of the compositions or methods provided herein, the ionizable lipid is D0G-IM4 (N- [ (25Z) -14- [ ( 9Z) -9-octadecen-l-yloxy ] -3, 6, 9, 12, 16-pentaoxatetratriacont-25-en-l-yl ] - 1 H-imi da z ole -5 -carboxamide) or analog. In an embodiment of any one of the compositions or methods provided herein, the ionizable lipid is OF-Deg-Lin ( 9Z, 12Z-octadecadienoic acid, 1, 1 ' , 1 ' ' , 1 ' ' ' - [ (3, 6-dioxo-2, 5-piperazinediyl) bis (4 , l-butanediylnitrilodi-2 , 1-ethanediyl) ] ester) or analog . In an embodiment of any one of the compositions or methods provided herein, the ionizable lipid is 0F-C4-Deg-Lin ( 9, 12-octadecadienoic acid, ( 9Z, 12Z) -1, 1 ' , 1 ' ' , 1 ' ' '- [ (3, 6-dioxo-2 , 5-piperazinediyl) bis (4 , l-butanediylnitrilodi-4 , 1-butanediyl) ] ester) or analog. In an embodiment of any one of the compositions or methods provided herein, the ionizable lipid is ATX-0114 (heptadecan-9-yl ( Z) -N- ( ( ( 4- (dimethylamino) butyl) thio) carbonyl) -N- (2- (non-2-en-l-yloxy) -2-oxoethyl) glycinate) or analog. In an embodiment of any one of the compositions or methods provided herein, the ionizable lipid is L202 ( l-methyl-4-piperidinecarboxylic acid, 2- [ 9-oxo-9- [ (3-pentyloctyl) oxy] nonyl] dodecyl ester) or analog . In an embodiment of any one of the compositions or methods provided herein, the ionizable lipid is C13-112-tetra-tail (14, 23-bis (2-hydroxytridecyl) -17, 20-dioxa-14, 23-diazahexatriacontane-12, 25-diol) or analog . In an embodiment of any one of the compositions or methods provided herein, the ionizable lipid is OC2-K3-E10 (3, 31- ( (2-hydroxyethyl) azanediyl) bis (N- (3- (bis (2-hydroxydecyl) amino) propyl) propanamide) or analog . In an embodiment of any one of the compositions or methods provided herein, the ionizable lipid is RCB-4-8 (bis ( (Z) -12- ( ( (non-2-yn-l-yloxy) carbonyl) oxy) octadec-9-en-l-yl) 3, 3' - ( (2- (diethylamino) ethyl) azanediyl) dipropionate) or analog. In an embodiment of any one of the compositions or methods provided herein, the ionizable lipid is RM 133-3 ( ( 9Z, 91Z) - ( (3, 31- ( (3- (diethylamino) propyl) azanediyl) bis (propanoyl) ) bis (oxy) ) bis (octadec-9-ene-18, 7-diyl) ( 5Z, 5 ' Z, 8Z, 8 ' Z, 11Z, 11,Z, 14Z, 14,Z, 17Z, 17,Z) -bis (icosa- 5, 8, 11, 14, 17-pentaenoate) ) or analog . In an embodiment of any one of the compositions or methods provided herein, the ionizable lipid is RM 137-5 ( ( 9Z , 9 ' Z ) - ( (3, 3 ' - ( (2- ( 4-methylpiperazin-l-yl) ethyl) azanediyl) bis (propanoyl) ) bis (oxy) ) bis (octadec-9-ene-18, 7-diyl) (4Z, 4 ' Z, 7Z, 7 ' Z, 10Z, 10 ' Z, 13Z, 13 ' Z, 16Z, 16 ' Z, 19Z, 19 ' Z) -bis (docosa-4 , 7, 10 , 13, 16, 19-hexaenoate) ) or analog . In an embodiment of any one of the compositions or methods provided herein, the ionizable lipid is PPZ-A10 (N,N ' - (piperazine-1, 4-diylbis (propane- 3, 1-diyl) ) bis (3- (didecylamino) propanamide) ) or analog. In an embodiment of any one of the compositions or methods provided herein, the ionizable lipid is Lipid 10 ( 4-methyl-l-piperazinepropanoic acid, 2- [di- ( 9Z, 12Z) -9, 12-octadecadien-l-ylamino ] ethyl ester) or analog. In an embodiment of any one of the compositions or methods provided herein, the ionizable lipid is Lipid AX4 or analog . In an embodiment of any one of the compositions or methods provided herein, the ionizable lipid is Lipid 5 (8- [ (2-hydroxyethyl) [8- (nonyloxy) -8-oxooctyl] amino] -octanoic acid, 1-octylnonyl ester) or analog. In an embodiment of any one of the compositions or methods provided herein, the ionizable lipid is TCL053 (2- ( ( ( 4- (dimethylamino) butanoyl) oxy) methyl) -2- ( ( ( ( Z) -tetradec-9-enoyl) oxy) methyl) propane-1, 3-diyl ( 9Z, 9 ' Z) -bis (tetradec-9-enoate) ) or analog . In an embodiment of any one of the compositions or methods provided herein, the ionizable lipid is YSK05 ( l-methyl-4 , 4-bis [ ( 9Z, 12Z) -9, 12-octadecadien-l-yloxy] -piperidine) or analog . In an embodiment of any one of the compositions or methods provided herein, the ionizable lipid is L-319 ( 9- [4- (dimethylamino) -1-oxobutoxy] -heptadecanedioic acid, 1, 17-di- (2Z) -2-nonen-l-yl ester) or analog. In an embodiment of any one of the compositions or methods provided herein, the ionizable lipid is 4A3-SC8 or analog . In an embodiment of any one of the compositions or methods provided herein, the ionizable lipid is C14-4 ( 1, 1 [ [2- [2- [4- [2- [ [2- [2- [bis (2-hydroxytetradecyl) amino] ethoxy] ethyl] (2-hydroxytetradecyl) amino] ethyl] -1-piperazinyl ] ethoxy] ethyl] imino] bis-2-tetradecanol) or analog. In an embodiment of any one of the compositions or methods provided herein, the ionizable lipid is DODMA ( 1, 2-dioleyloxy-3-dimethylaminopropane) or analog. In an embodiment of any one of the compositions or methods provided herein, the ionizable lipid is DOBAQ (N- ( 4-carboxybenzyl ) -N, N-dimethyl-2 , 3-bis (oleoyloxy) propan-l-aminium) or analog. In an embodiment of any one of the compositions or methods provided herein, the ionizable lipid is a cationic amino lipid. In an embodiment of any one of the compositions or methods provided herein, the ionizable lipid is DLin-DMA (N, N-dimethyl-2 , 3-bis [ ( 9Z, 12Z) -9, 12-octadecadien-l-yloxy] -1-propanamine) or analog . In an embodiment of any one of the compositions or methods provided herein, the ionizable lipid is TT3 (Nl, N3,N5-tris [3- (didodecylamino) propyl] -1, 3, 5 -benzenetri carboxamide) or analog. In an embodiment of any one of the compositions or methods provided herein, the ionizable lipid is FTT5 (Hexa (octan-3-yl)

[0016] 9, 9 ' , 9", 9"', 9"", 9""'- ( ( ( (benzene-1, 3, 5-tricarbonyl) ris (azanediyl) ) tris (propane-3, 1-diyl) ) tris (azanetriyl) ) hexanonanoate) . In an embodiment of any one of the compositions or methods provided herein, the ionizable lipid is OF-02 ( 3, 6-bis [ 4- [bis [ ( 9Z , 12Z ) -2-hydroxy-9, 12-octadecadien-l-yl] amino] butyl] -2 , 5-piperazinedione) or analog . In an embodiment of any one of the compositions or methods provided herein, the ionizable lipid is AA3-DLin (piperazine-1, 4-diylbis (ethane-2 , 1-diyl) ( 9Z, 91Z, 12Z, 121Z) -bis (octadeca-9, 12-dienoate) ) or analog . In an embodiment of any one of the compositions or methods provided herein, the ionizable lipid is DLin-MC3-DMA (4- (dimethylamino) -butanoic acid, ( 10Z, 13Z) -1- ( 9Z, 12Z) -9, 12-octadecadien-l-yl-10 , 13-nonadecadien-l-yl ester) or analog . In an embodiment of any one of the compositions or methods provided herein, the ionizable lipid is DLin-KC2-DMA (N, N-dimethyl-2 , 2 -di- ( 9Z, 12Z) -9, 12-octadecadien-l-yl-l, 3-dioxolane-4-ethanamine) or analog. In an embodiment of any one of the compositions or methods provided herein, the ionizable lipid is SM-102 ( 8— [ (2— hydroxyethyl) [ 6-oxo-6- (undecyloxy) hexyl] amino] -octanoic acid, 1-octylnonyl ester) or analog. In an embodiment of any one of the compositions or methods provided herein, the ionizable lipid is ALC-0315 ( 6- ( ( 2-hexyldecanoyl) oxy) -N- ( 6- ( ( 2-hexyldecanoyl) oxy) hexyl) -N- ( 4-hydroxybutyl) hexan-l-aminium) or analog. In an embodiment of any one of the compositions or methods provided herein, the ionizable lipid is ALC-0315 analogous-1 ( ( (2-hydroxyethyl) azanediyl) bis (octane-8 , 1-diyl) bis (2-hexyldecanoate) ) or analog . In an embodiment of any one of the compositions or methods provided herein, the ionizable lipid is Lipid 222 ( ( (4-hydroxybutyl) azanediyl) bis (octane-8 , 1-diyl) bis (2-hexyldecanoate) ) or analog . In an embodiment of any one of the compositions or methods provided herein, the ionizable lipid is CL4-H6 ( 9 -octadecenoic acid, 1, 1 ' - [ 7- [4- (dipropyl amino) butyl] -7-hydroxy-l , 13-tridecanediyl] ester) or analog. In an embodiment of any one of the compositions or methods provided herein, the ionizable lipid is AL-A12 (N-dodecyl-N- ( 3-methoxypropyl) dodecan-l-amine) or analog. In an embodiment of any one of the compositions or methods provided herein, the ionizable lipid is Lipid 14 ( ( (2- ( (4- ( dimethylamino) butanoyl) oxy) ethyl) azanediyl) bis (octane-8, 1-diyl) bis (2-hexyldecanoate) ) or analog . In an embodiment of any one of the compositions or methods provided herein, the ionizable lipid is Lipid 16 ( ( (2- ( ( 4- (dimethylamino) butanoyl) oxy) ethyl) azanediyl) bis (octane-8 , 1-diyl) bis (2-hexyldecanoate) ) or analog . In an embodiment of any one of the compositions or methods provided herein, the ionizable lipid is a cationic alkyne lipid. In an embodiment of any one of the compositions or methods provided herein, the ionizable lipid is Lipid A6 (di (dec-3-yn-l-yl) 9- ( ( 4- (dimethylamino) butanoyl) oxy) heptadecanedioate) or analog. In an embodiment of any one of the compositions or methods provided herein, the ionizable lipid is a cationic trialkyl lipid. In an embodiment of any one of the compositions or methods provided herein, the ionizable lipid is Lipid CL1 ( 5- (dimethylamino) -pentanoic acid, ( 6Z) -l, 2-di- ( 4Z) -4-decen-l-yl-6-dodecen-l-yl ester) or analog. In an embodiment of any one of the compositions or methods provided herein, the ionizable lipid is a disulfide-bond containing cationic lipid. In an embodiment of any one of the compositions or methods provided herein, the ionizable lipid is a BAMEA-016B (bis (2- (dodecyldisulf aneyl) ethyl) 3, 31- ( (3-methyl-9-oxo-10-oxa-13, 14-dithia-3, 6-diazahexacosyl) azanediyl) dipropionate) or analog . In an embodiment of any one of the compositions or methods provided herein, the ionizable lipid is Coatsome SS-E (Bis { 2- [N-methyl-N- (a-D-tocopherolhemisuccinatepropyl) amino] ethyl } disulf ide) or analog. In an embodiment of any one of the compositions or methods provided herein, the ionizable lipid is Coatsome SS-EC (Bis { 2- [ 4- (a-D-tocopherolhemisuccinateethyl) piperidyl] ethyl } disulfide) or analog . In an embodiment of any one of the compositions or methods provided herein, the ionizable lipid is Coatsome SS-OB (is Coatsome SS-OC (Bis { 2- [ 4- (cis-9-octadecenoateethyl) -1-piperidinyl ] ethyl } disulf ide) or analog . In an embodiment of any one of the compositions or methods provided herein, the ionizable lipid is Coatsome SS-OB (Bis [2- (4-{ 2- [ 4- ( cis-9-octadecenoyloxy) phenylacetoxy] ethyl }piperidinyl) ethyl] disulfide) or analog . In an embodiment of any one of the compositions or methods provided herein, the ionizable lipid is a lipomer . In an embodiment of any one of the compositions or methods provided herein, the ionizable lipid is CKK-E12 (3, 6-bis [ 4- [bis (2-hydroxydodecyl) amino] butyl] -2, 5-piperazinedione) or analog. In an embodiment of any one of the compositions or methods provided herein, the ionizable lipid is a lipidoid or lipidoid analog. In an embodiment of any one of the compositions or methods provided herein, the ionizable lipid is C12-113 ( 1, 1 ' , 1 1 ' ' ' - [ (methylimino) bis (2, 1-ethanediylnitrilo) ] tetrakis-2-dodecanol) or analog . In an embodiment of any one of the compositions or methods provided herein, the ionizable lipid is C12-200 ( 1, 1 '- [ [2- [4- [2- [ [2- [bis (2-hydroxydodecyl) amino] ethyl] (2-hydroxydodecyl) amino] ethyl] -1-piperazinyl] ethyl] imino] bis-2-dodecanol) or analog . In an embodiment of any one of the compositions or methods provided herein, the ionizable lipid is NT1-014B or analog. In an embodiment of any one of the compositions or methods provided herein, the ionizable lipid is 306-O12B-3 (bis (2- (octyldisulfaneyl) ethyl) 3 , 31- ( ( 4-methyl-ll-oxo-12-oxa-15, 16-dithia-4, 8-diazatetracosyl) azanediyl) dipropionate) or analog. In an embodiment of any one of the compositions or methods provided herein, the ionizable lipid 503-013 or analog. In an embodiment of any one of the compositions or methods provided herein, the ionizable lipid is a disulfide-cleavable proton-activated lipid like material . In an embodiment of any one of the compositions or methods provided herein, the ionizable lipid is SS-Balm-M (tetradecanoic acid, 1, 1 ' - [dithiobis [2 , 1-ethanediyl (methylimino) -3, 1- propanediyl] ] ester) or analog . In an embodiment of any one of the compositions or methods provided herein, the ionizable lipid is SS-PalmO-Phe ( 4- [ [ ( 9Z ) -l-oxo-9-octadecen-l-yl ] oxy] -benzeneacetic acid, 1, 1 '- [dithiobis (2, 1-ethanediyl-l , 4-piperidinediyl-2 , 1-ethanediyl) ] ester) or analog .

[0017] In an embodiment of any one of the compositions or methods provided herein, the structural lipid is a saturated phosphatidic acid. In an embodiment of any one of the compositions or methods provided herein, the structural lipid is DMPA ( 1, 2-dimyristoyl-sn-glycero-3-phosphate) or analog. In an embodiment of any one of the compositions or methods provided herein, the structural lipid is DPPA ( 1 , 2-dipalmitoyl-sn-glycero-3-phosphate) or analog . In an embodiment of any one of the compositions or methods provided herein, the structural lipid is DSPA ( 1, 2-distearoyl-sn-glycero-3-phosphate) or analog. In an embodiment of any one of the compositions or methods provided herein, the structural lipid is DLPA ( 1 , 2-dilauroyl-sn-glycero-3-phosphate) or analog. In an embodiment of any one of the compositions or methods provided herein, the structural lipid is an unsaturated phosphatidic acid. In an embodiment of any one of the compositions or methods provided herein, the structural lipid is DOPA ( 1, 2-dioleoyl-sn-glycero-3-phosphate) or analog . In an embodiment of any one of the compositions or methods provided herein, the structural lipid is DAPA ( 1 , 2-diarachidonoyl-sn-glycero-3-phosphate) or analog . In an embodiment of any one of the compositions or methods provided herein, the structural lipid is a phosphatidic acid with both saturated and unsaturated tails . In an embodiment of any one of the compositions or methods provided herein, the structural lipid is POPA ( l-palmitoyl-2-oleoyl-sn-glycero-3-phosphate) or analog . In an embodiment of any one of the compositions or methods provided herein, the structural lipid is PAPA ( 1-palmitoyl-2-arachidonoyl-sn-glycero-3-phosphate) or analog . In an embodiment of any one of the compositions or methods provided herein, the structural lipid is SOPAPA (l-stearoyl-2-oleoyl-sn-glycero-3-phosphate) or analog . In an embodiment of any one of the compositions or methods provided herein, the structural lipid is SAPA ( l-stearoyl-2- arachidonoyl-sn-glycero-3-phosphate) or analog . In an embodiment of any one of the compositions or methods provided herein, the structural lipid is a saturated phosphatidylcholine . In an embodiment of any one of the compositions or methods provided herein, the structural lipid is DHPC ( 1 , 2-dihexanoyl-sn-glycero-3-phosphocholine or 1, 2-diheptanoyl-sn-glycero-3-phosphocholine) or analog . In an embodiment of any one of the compositions or methods provided herein, the structural lipid is DLPC ( 1, 2-dilauroyl-sn-glycero-3-phosphocholine) or analog. In an embodiment of any one of the compositions or methods provided herein, the structural lipid is DMPC ( 1 , 2-dimyristoyl-sn-glycero-3-phosphocholine) or analog. In an embodiment of any one of the compositions or methods provided herein, the structural lipid is DPPC ( 1, 2-dipalmitoyl-sn-glycero-3-phosphocholine) or analog. In an embodiment of any one of the compositions or methods provided herein, the structural lipid is DSPC ( 1, 2-distearoyl-sn-glycero-3-phosphocholine) or analog. In an embodiment of any one of the compositions or methods provided herein, the structural lipid is an unsaturated phosphatidylcholine . In an embodiment of any one of the compositions or methods provided herein, the structural lipid is DOPC ( 1 , 2-dioleoyl-sn-glycero-3-phosphocholine) or analog. In an embodiment of any one of the compositions or methods provided herein, the structural lipid is DLPC ( 1, 2-dilinoleoyl-sn-glycero-3-phosphocholine) or analog. In an embodiment of any one of the compositions or methods provided herein, the structural lipid is DEPC ( 1 , 2-dierucoyl-sn-glycero-3-phosphocholine) or analog. In an embodiment of any one of the compositions or methods provided herein, the structural lipid is 1, 2-DAPC ( 1, 2-Diarachidoyl-sn-glycero-3-phosphocholine) or analog. In an embodiment of any one of the compositions or methods provided herein, the structural lipid is DDPC ( 1, 2-Didecanoyl-sn-glycero-3-phosphocholine) or analog. In an embodiment of any one of the compositions or methods provided herein, the structural lipid is 1, 2-PLPC ( l-Palmitoyl-2-Lauroyl-sn-glycero-3-Phosphocholine) or analog . In an embodiment of any one of the compositions or methods provided herein, the structural lipid is an unsaturated phosphatidylcholine with tails of different numbers of carbon atoms . In an embodiment of any one of the compositions or methods provided herein, the structural lipid is MPPC (l-myristoyl-2-palmitoyl-sn-glycero-3-phosphocholine) or analog . In an embodiment of any one of the compositions or methods provided herein, the structural lipid is MSPC ( l-myristoyl-2-stearoyl-sn-glycero-3-phosphocholine) or analog . In an embodiment of any one of the compositions or methods provided herein, the structural lipid is PMPC ( l-palmitoyl-2-myristoyl-sn-glycero-3-phosphocholine) or analog . In an embodiment of any one of the compositions or methods provided herein, the structural lipid is PSPC ( l-palmitoyl-2-stearoyl-sn-glycero-3-phosphocholine) or analog. In an embodiment of any one of the compositions or methods provided herein, the structural lipid is SMPC ( l-stearoyl-2-myristoyl-sn-glycero-3-phosphocholine) or analog. In an embodiment of any one of the compositions or methods provided herein, the structural lipid is SPPC ( l-stearoyl-2-palmitoyl-sn-glycero-3-phosphocholine) or analog. In an embodiment of any one of the compositions or methods provided herein, the structural lipid is a phosphatidylcholine with both saturated and unsaturated tails . In an embodiment of any one of the compositions or methods provided herein, the structural lipid is POPC ( l-palmitoyl-2-oleoyl-glycero-3-phosphocholine) or analog. In an embodiment of any one of the compositions or methods provided herein, the structural lipid is PAPC ( l-palmitoyl-2-arachidonoyl-sn-glycero-3-phosphocholine) or analog . In an embodiment of any one of the compositions or methods provided herein, the structural lipid is SOPC ( l-stearoyl-2-oleoyl-sn-glycero-3-phosphocholine) or analog. In an embodiment of any one of the compositions or methods provided herein, the structural lipid is OMPC ( l-oleoyl-2-myristoyl-sn-glycero-3-phosphocholine) or analog. In an embodiment of any one of the compositions or methods provided herein, the structural lipid is OPPC ( l-oleoyl-2-palmitoyl-sn-glycero-3-phosphocholine) or analog. In an embodiment of any one of the compositions or methods provided herein, the structural lipid is OSPC ( l-oleoyl-2-stearoyl-sn-glycero-3-phosphocholine) or analog . In an embodiment of any one of the compositions or methods provided herein, the structural lipid is a lysophosphatidylcholine . In an embodiment of any one of the compositions or methods provided herein, the structural lipid is S-LysoPC (l-Stearoyl-2-lyso-sn-glycero-3-phosphocholine) or analog . In an embodiment of any one of the compositions or methods provided herein, the structural lipid is P-LysoPC ( l-Palmitoyl-2-lyso-sn-glycero-3-phosphocholine) or analog . In an embodiment of any one of the compositions or methods provided herein, the structural lipid is O-LysoPC ( l-01eoyl-2-lyso-sn-glycero-3-phosphocholine) or analog . In an embodiment of any one of the compositions or methods provided herein, the structural lipid is M-LysoPC ( l-Myristoyl-2-lyso-sn-glycero-3-phosphocholine) or analog. In an embodiment of any one of the compositions or methods provided herein, the structural lipid is a saturated phosphatidylethanolamine . In an embodiment of any one of the compositions or methods provided herein, the structural lipid is DLPE ( 1 , 2-dilauroyl-sn-glycero-3-phosphoethanolamine) or analog. In an embodiment of any one of the compositions or methods provided herein, the structural lipid is DPPE ( 1, 2-dipalmitoyl-sn-glycero-3-phosphoethanolamine) or analog . In an embodiment of any one of the compositions or methods provided herein, the structural lipid is DPPE-Mal (N- ( 3-Maleimide-l-oxopropyl) -L-a-phosphatidylethanolamine, Dipalmitoyl) or analog . In an embodiment of any one of the compositions or methods provided herein, the structural lipid is DPPE-NHS (N- (Succinimidyloxy-glutaryl) -L-a-phosphatidylethanolamine, Dipalmitoyl) or analog . In an embodiment of any one of the compositions or methods provided herein, the structural lipid is DPPE-Glu (N-Glutaryl-L-a-phosphatidylethanolamine, Dipalmitoyl) or analog . In an embodiment of any one of the compositions or methods provided herein, the structural lipid is DSPE ( 1, 2-distearoyl-sn-glycero-3-phosphoethanolamine) or analog . In an embodiment of any one of the compositions or methods provided herein, the structural lipid is DSPE-Mal (N- (3-Maleimide-l-oxopropyl) -L-a-phosphatidylethanolamine, Distearoyl) or analog. In an embodiment of any one of the compositions or methods provided herein, the structural lipid is DSPE-NHS (N- ( Succinimidyloxy-glutaryl) -L-a-phosphatidy let hanol amine, Distearoyl) or analog. In an embodiment of any one of the compositions or methods provided herein, the structural lipid is DSPE-Glu (N-Glutaryl-L-a-phosphatidylethanolamine, Distearoyl) or analog. In an embodiment of any one of the compositions or methods provided herein, the structural lipid is DMPE ( 1 , 2-dimyristoyl-sn-glycero-3-phosphoethanolamine) or analog . In an embodiment of any one of the compositions or methods provided herein, the structural lipid is DMPE-Mal (N- (3-Maleimide-l-oxopropyl) -L-a-phosphatidylethanolamine, Dimyristoyl) or analog. In an embodiment of any one of the compositions or methods provided herein, the structural lipid is DMPE-NHS (N- (Succinimidyloxy-glutaryl) -L-a-phosphatidylethanolamine, Dimyristoyl) or analog . In an embodiment of any one of the compositions or methods provided herein, the structural lipid is DMPE-Glu (N-Glutaryl-L-a-phosphatidylethanolamine, Dimyristoyl) or analog. In an embodiment of any one of the compositions or methods provided herein, the structural lipid is an unsaturated phosphatidylethanolamine . In an embodiment of any one of the compositions or methods provided herein, the structural lipid is DOPE ( 1 , 2-dioleoyl-sn-glycero-3-phosphoethanolamine) or analog. In an embodiment of any one of the compositions or methods provided herein, the structural lipid is DOPE-Mal (N- (3-Maleimide-l-oxopropyl) -L-a-phosphatidylethanolamine, Dioleoyl) or analog. In an embodiment of any one of the compositions or methods provided herein, the structural lipid is DOPE-NHS (N- ( Succinimidyloxy-glutaryl ) -L-a-phosphatidylethanolamine, Dioleoyl) or analog. In an embodiment of any one of the compositions or methods provided herein, the structural lipid is DOPE-Glu (N-Glutaryl-L-a-phosphatidylethanolamine, Dioleoyl) or analog . In an embodiment of any one of the compositions or methods provided herein, the structural lipid is DAPE ( 1 , 2-diarachidonoyl-sn-glycero-3-phosphoethanolamine) or analog . In an embodiment of any one of the compositions or methods provided herein, the structural lipid is a phosphatidylethanolamine with both saturated and unsaturated tails . In an embodiment of any one of the compositions or methods provided herein, the structural lipid is POPE ( l-palmitoyl-2-oleoyl-sn-glycero-3-phosphoethanolamine) or analog. In an embodiment of any one of the compositions or methods provided herein, the structural lipid is SOPE ( l-stearoyl-2-oleoyl-sn-glycero-3-phosphoethanolamine) or analog . In an embodiment of any one of the compositions or methods provided herein, the structural lipid is PAPE ( l-palmitoyl-2- arachidonoyl-sn-glycero-3-phosphoethanolamine) or analog. In an embodiment of any one of the compositions or methods provided herein, the structural lipid is SAFE ( l-stearoyl-2-arachidonoyl-sn-glycero-3-phosphoethanolamine) or analog . In an embodiment of any one of the compositions or methods provided herein, the structural lipid is a saturated phosphatidylglycerol . In an embodiment of any one of the compositions or methods provided herein, the structural lipid is DLPG ( 1 , 2-dilauroyl-sn-glycero-3-phospho- ( 1 ' -rac-glycerol) ) or analog . In an embodiment of any one of the compositions or methods provided herein, the structural lipid is DMPG ( 1, 2-dimyristoyl-sn-glycero-3-phospho- (1 ' -rac-glycerol) ) or analog . In an embodiment of any one of the compositions or methods provided herein, the structural lipid is DP PG ( 1, 2 -dipalmitoyl-sn-glycer o-3 -phospho- ( 1 ' -rac-glycerol) ) or analog . In an embodiment of any one of the compositions or methods provided herein, the structural lipid is DSPG ( 1 , 2-distearoyl-sn-glycero-3-phospho- (1 ' -rac-glycerol) ) or analog . In an embodiment of any one of the compositions or methods provided herein, the structural lipid is DOPG ( 1 , 2-dioleoyl-sn-glycero-3-phospho- ( 1 ' -rac-glycerol) ) or analog . In an embodiment of any one of the compositions or methods provided herein, the structural lipid is an unsaturated phosphatidylglycerol . In an embodiment of any one of the compositions or methods provided herein, the structural lipid is a phosphatidylglycerol with both saturated and unsaturated tails . In an embodiment of any one of the compositions or methods provided herein, the structural lipid is POPG ( l-palmitoyl-2-oleoyl-sn-glycero-3-phospho- (1 ' -rac-glycerol) ) or analog . In an embodiment of any one of the compositions or methods provided herein, the structural lipid is PAPG ( l-palmitoyl-2-arachidonoyl-sn-glycero-3-phospho- ( 1 ' -rac-glycerol) ) or analog . In an embodiment of any one of the compositions or methods provided herein, the structural lipid is SOPG (1-stearoyl-2-oleoyl-sn-glycero-3-phospho- (1 ' -rac-glycerol) ) or analog. In an embodiment of any one of the compositions or methods provided herein, the structural lipid is SAPG ( l-stearoyl-2-arachidonoyl-sn-glycero-3-phospho- (1 ' -rac-glycerol) ) or analog . In an embodiment of any one of the compositions or methods provided herein, the structural lipid is a saturated phosphatidylserine . In an embodiment of any one of the compositions or methods provided herein, the structural lipid is DSPS ( 1 , 2-distearoyl-sn-glycero-3-phospho-L-serine) or analog. In an embodiment of any one of the compositions or methods provided herein, the structural lipid is DMPS ( 1, 2-dimyristoyl-sn-glycero-3-phospho-L-serine) or analog. In an embodiment of any one of the compositions or methods provided herein, the structural lipid is DPPS ( 1, 2-dipalmitoyl-sn-glycero-3-phospho-L-serine) or analog. In an embodiment of any one of the compositions or methods provided herein, the structural lipid is DLPS ( 1, 2-dilauroyl-sn-glycero-3-phospho-L-serine) or analog. In an embodiment of any one of the compositions or methods provided herein, the structural lipid is an unsaturated phosphatidylserine . In an embodiment of any one of the compositions or methods provided herein, the structural lipid is DOPS ( 1, 2-dioleoyl-sn-glycero-3-phospho-L-serine) or analog . In an embodiment of any one of the compositions or methods provided herein, the structural lipid is a phosphatidylserine with both saturated and unsaturated tails . In an embodiment of any one of the compositions or methods provided herein, the structural lipid is POPS ( l-palmitoyl-2-oleoyl-sn-glycero-3-phospho-L-serine) or analog. In an embodiment of any one of the compositions or methods provided herein, the structural lipid is SOPS ( l-stearoyl-2-oleoyl-sn-glycero-3-phospho-L-serine) or analog . In an embodiment of any one of the compositions or methods provided herein, the structural lipid is PAPS ( l-palmitoyl-2-arachidonoyl-sn-glycero-3-phospho-L-serine) or analog. In an embodiment of any one of the compositions or methods provided herein, the structural lipid is SAPS ( 1-st ear oy 1-2 -ar achidonoyl-sn-glycero-3-phospho-L- serine) or analog .

[0018] In an embodiment of any one of the compositions or methods provided herein, the cationic lipid is DORI (N- (2-hydroxyethyl) -N, N-dimethyl-2 , 3-bis (oleoyloxy) propan-l-aminium bromide) or analog . In an embodiment of any one of the compositions or methods provided herein, the cationic lipid is

[0019] DC-6-14 (0, 0' -ditetradecanoyl-N- (a-trimethylammonioacetyl) diethanolamine) or analog . In an embodiment of any one of the compositions or methods provided herein, the cationic lipid is

[0020] DSDMA ( 1 , 2-distearyloxy-N, N-dimethyl-3-aminopropane) or analog . In an embodiment of any one of the compositions or methods provided herein, the cationic lipid is DMRIE ( 1 , 2-dimyristyloxypropyl-3-dimethyl-hydroxyethyl ammonium bromide) or analog . In an embodiment of any one of the compositions or methods provided herein, the cationic lipid is a quaternary ammonium compound. In an embodiment of any one of the compositions or methods provided herein, the cationic lipid is 18 : 0 DDAB (Dimethyldioctadecylammonium) or analog. In an embodiment of any one of the compositions or methods provided herein, the cationic lipid is DC-1-16 (N-Hexadecyl-N, N-dimethylhexadecan-l-aminium) or analog . In an embodiment of any one of the compositions or methods provided herein, the cationic lipid is dimethyldimyristylammonium (N, N-dimethyl-N-tetradecyl-l-tetradecanaminium) or analog . In an embodiment of any one of the compositions or methods provided herein, the cationic lipid is DODAC N, N-Dioleyl-N, N-dimethylammonium chloride) or analog. In an embodiment of any one of the compositions or methods provided herein, the cationic lipid is a trimethylammoniumpropane . In an embodiment of any one of the compositions or methods provided herein, the cationic lipid is 14 : 0 TAP ( 1, 2-dimyristoyl-3-trimethylammoniumpropane) or analog. In an embodiment of any one of the compositions or methods provided herein, the cationic lipid is 16 : 0 TAP ( 1, 2-dipalmitoyl-3-trimethylammoniumpropane) or analog. In an embodiment of any one of the compositions or methods provided herein, the cationic lipid is 18 : 0 TAP ( 1, 2-stearoyl-3-trimethylammoniumpropane) or analog. In an embodiment of any one of the compositions or methods provided herein, the cationic lipid is 18 : 1 TAP (DOTAP, 1 , 2-dioleoyl-3-trimethylammoniumpropane) or analog. In an embodiment of any one of the compositions or methods provided herein, the cationic lipid is DOTMA ( 1 , 2-di-0-octadecenyl-3-trimethylammonium propane) or analog . In an embodiment of any one of the compositions or methods provided herein, the cationic lipid is an esterified phosphatidylcholine . In an embodiment of any one of the compositions or methods provided herein, the cationic lipid is 12 : 0 EPC ( 1 , 2-dilauroyl-sn-glycero-3-ethylphosphocholine) or analog . In an embodiment of any one of the compositions or methods provided herein, the cationic lipid is 14 : 0 EPC ( 1, 2-dimyristoyl-sn-glycero-3-ethylphosphocholine) or analog . In an embodiment of any one of the compositions or methods provided herein, the cationic lipid is 16 : 0 EPC (1, 2-dipalmitoyl-sn-glycero-3-ethylphosphocholine) or analog . In an embodiment of any one of the compositions or methods provided herein, the cationic lipid is 18 : 0 EPC ( 1, 2-distearoyl-sn-glycero-3-ethylphosphocholine) or analog . In an embodiment of any one of the compositions or methods provided herein, the cationic lipid is 14 : 1 EPC ( 1 , 2-dimyristoleoyl-sn-glycero-3-ethylphosphocholine) or analog. In an embodiment of any one of the compositions or methods provided herein, the cationic lipid is 18 : 1 EPC ( 1, 2-dioleoyl-sn-glycero-3-ethylphosphocholine) or analog . In an embodiment of any one of the compositions or methods provided herein, the cationic lipid is 16 : 0-18 : 1 EPC ( l-palmitoyl-2-oleoyl-sn-glycero-3-ethylphosphocholine) or analog . In an embodiment of any one of the compositions or methods provided herein, the cationic lipid is a multivalent cationic lipid. In an embodiment of any one of the compositions or methods provided herein, the cationic lipid is MVL5 (Nl- [2- ( ( IS) -1- [ (3-aminopropyl) amino] -4- [di (3-amino-propyl) amino] butylcarboxamido) ethyl] -3, 4-di [oleyloxy] -benzamide) or analog. In an embodiment of any one of the compositions or methods provided herein, the cationic lipid is a spermine-derived lipid. In an embodiment of any one of the compositions or methods provided herein, the cationic lipid is DOSPA (2 , 3-dioleyloxy-N- [2- ( sperminecarboxamido) ethyl] -N, N-dimethyl-1-propanaminium) or analog . In an embodiment of any one of the compositions or methods provided herein, the cationic lipid is GL67 (N4-Cholesteryl-Spermine) or analog. In an embodiment of any one of the compositions or methods provided herein, the cationic lipid is DOGS (Dioctadecylamidoglycylspermine) or analog. In an embodiment of any one of the compositions or methods provided herein, the cationic lipid is DPPES (Dipalmitoylphosphatidylethanolamidospermine) or analog . In an embodiment of any one of the compositions or methods provided herein, the cationic lipid is RPR120535 or analog. In an embodiment of any one of the compositions or methods provided herein, the cationic lipid is a cholesterol-based cationic lipid. In an embodiment of any one of the compositions or methods provided herein, the cationic lipid is DC-Cholesterol • HC1 (3f>- [N- (N ' , N ' -dimethylaminoethane) -carbamoyl ] cholesterol hydrochloride) or analog. In an embodiment of any one of the compositions or methods provided herein, the cationic lipid is CLin-DMA ( 3-dimethylamino-2- ( cholest-5-en-3-beta-oxybutan-4-oxy) -1- (cis, cis-9, 12-octadecadienoxy) propane) or analog .

[0021] In an embodiment of any one of the compositions or methods provided herein, the cationic or ionizable substance is comprised of a cationic polymer, block copolymer, copolymer, alternating copolymer, or random copolymer such as polyethyleneimine, poly (amidoamine) , or its derivative .

[0022] In an embodiment of any one of the compositions or methods provided herein, the polymer is a polyethylene glycol (PEG) -modified lipid. In an embodiment of any one of the compositions or methods provided herein, the polymer is a PEG-modified lipid with a polyethylene glycol chain of up to 350 Da in length . In an embodiment of any one of the compositions or methods provided herein, the polymer is 14 : 0 PEG350 PE ( 1 , 2-dimyristoyl-sn-glycero-3-phosphoethanolamine-N- [methoxy (polyethylene glycol) -350 ] ) or analog. In an embodiment of any one of the compositions or methods provided herein, the polymer is 16 : 0 PEG350 PE ( 1, 2-dipalmitoyl-sn-glycero-3-phosphoethanolamine-N- [methoxy (polyethylene glycol) -350 ] ) or analog. In an embodiment of any one of the compositions or methods provided herein, the polymer is 18 : 0 PEG350 PE ( 1, 2-distearoyl-sn-glycero-3-phosphoethanolamine-N- [methoxy (polyethylene glycol) -350 ] ) or analog. In an embodiment of any one of the compositions or methods provided herein, the polymer is 18 : 1 PEG350 PE ( 1, 2-dioleoyl-sn-glycero-3-phosphoethanolamine-N- [methoxy (polyethylene glycol) -350 ] ) or analog. In an embodiment of any one of the compositions or methods provided herein, the polymer is a PEG-modified lipid with a polyethylene glycol chain of up to 550 Da in length . In an embodiment of any one of the compositions or methods provided herein, the polymer is 14 : 0 PEG550 PE (DMPE-PEG550 , 1, 2-dimyristoyl-sn-glycero-3-phosphoethanolamine-N- [methoxy (polyethylene glycol) -550 ] ) or analog. In an embodiment of any one of the compositions or methods provided herein, the polymer is 16 : 0 PEG550 PE ( 1 , 2-dipalmitoyl-sn-glycero-3-phosphoethanolamine-N- [methoxy (polyethylene glycol) -550 ] ) or analog. In an embodiment of any one of the compositions or methods provided herein, the polymer is 18 : 0 PEG550 PE (DSPE-PEG550, 1 , 2-distearoyl-sn-glycero-3-phosphoethanolamine-N- [methoxy (polyethylene glycol) -550 ] ) or analog. In an embodiment of any one of the compositions or methods provided herein, the polymer is 18 : 1 PEG550 PE (DOPE-PEG550 , 1 , 2-dioleoyl-sn-glycero-3-phosphoethanolamine-N- [methoxy (polyethylene glycol) -550 ] ) or analog . In an embodiment of any one of the compositions or methods provided herein, the polymer is a PEG-modified lipid with a polyethylene glycol chain of up to 750 Da in length . In an embodiment of any one of the compositions or methods provided herein, the polymer is 14 : 0 PEG750 PE ( 1 , 2-dimyristoyl-sn-glycero-3-phosphoethanolamine-N- [methoxy (polyethylene glycol) -750 ] ) or analog. In an embodiment of any one of the compositions or methods provided herein, the polymer is 16 : 0 PEG750 PE ( 1, 2-dipalmitoyl-sn-glycero-3-phosphoethanolamine-N- [methoxy (polyethylene glycol) -750 ] ) or analog. In an embodiment of any one of the compositions or methods provided herein, the polymer is 18 : 0 PEG750 PE ( 1, 2-distearoyl-sn-glycero-3-phosphoethanolamine-N- [methoxy (polyethylene glycol) -750 ] ) or analog. In an embodiment of any one of the compositions or methods provided herein, the polymer is 18 : 1 PEG750 PE ( 1, 2-dioleoyl-sn-glycero-3-phosphoethanolamine-N- [methoxy (polyethylene glycol) -750 ] ) or analog. In an embodiment of any one of the compositions or methods provided herein, the polymer is a PEG-modified lipid with a polyethylene glycol chain of up to 1 kDa in length . In an embodiment of any one of the compositions or methods provided herein, the polymer is 14 : 0 PEG1000 PE (DMPE-PEG1000 , 1, 2-dimyristoyl-sn-glycero-3-phosphoethanolamine-N- [methoxy (polyethylene glycol) -1000 ] ) or analog . In an embodiment of any one of the compositions or methods provided herein, the polymer is 16 : 0 PEG1000 PE (DPPE-PEG1000 , 1, 2-dipalmitoyl-sn-glycero-3-phosphoethanolamine-N- [methoxy (polyethylene glycol) -1000 ] ) or analog . In an embodiment of any one of the compositions or methods provided herein, the polymer is 18 : 0 PEG1000 PE (DSPE-PEG1000, 1 , 2-distearoyl-sn-glycero-3-phosphoethanolamine-N- [methoxy (polyethylene glycol) -1000 ] ) or analog . In an embodiment of any one of the compositions or methods provided herein, the polymer is 18 : 1 PEG1000 PE (DOPE-PEGIOOO, 1 , 2-dioleoyl-sn-glycero-3-phosphoethanolamine-N- [methoxy (polyethylene glycol) -1000 ] ) or analog. In an embodiment of any one of the compositions or methods provided herein, the polymer is a PEG-modified lipid with a polyethylene glycol chain of up to 2 kDa in length . In an embodiment of any one of the compositions or methods provided herein, the polymer is 14 : 0 PEG2000 PE (DMPE-PEG2000 , 1, 2-dimyristoyl-sn-glycero-3-phosphoethanolamine-N- [methoxy (polyethylene glycol) -2000 ] ) or analog . In an embodiment of any one of the compositions or methods provided herein, the polymer is 16 : 0 PEG2000 PE (DPPE-PEG2000, 1 , 2-dipalmitoyl-sn-glycero-3-phosphoethanolamine-N- [methoxy (polyethylene glycol) -2000 ] ) or analog . In an embodiment of any one of the compositions or methods provided herein, the polymer is 18 : 0 PEG2000 PE (DSPE-PEG2000, 1 , 2-distearoyl-sn-glycero-3-phosphoethanolamine-N- [methoxy (polyethylene glycol) -2000 ] ) or analog . In an embodiment of any one of the compositions or methods provided herein, the polymer is DSPE-2arm-PEG2000 (N- [2 ' , 3 ' -Bis (methylpolyoxyethylene oxy) propane-1 ' -oxycarbonyl] -1, 2-distearoyl-sn-glycero-3-phosphoethanolamine) or analog . In an embodiment of any one of the compositions or methods provided herein, the polymer is DSPE-PEG2K-triGalNAc ( 1 , 2-distearoyl-sn-glycero-3-phosphoethanolamine-N- [carboxy (polyethylene glycol) -2000 ] -N-tri (N-acetylgalactosamine) ) or analog. In an embodiment of any one of the compositions or methods provided herein, the polymer is 18 : 1 PEG2000 PE (DGPE-PEG2000, 1 , 2-dioleoyl-sn-glycero-3-phosphoethanolamine-N- [methoxy (polyethylene glycol) -2000 ] ) or analog . In an embodiment of any one of the compositions or methods provided herein, the polymer is a PEG-modified lipid with a polyethylene glycol chain of up to 3 kDa in length . In an embodiment of any one of the compositions or methods provided herein, the polymer is 14 : 0 PEG3000 PE ( 1, 2-dimyristoyl-sn-glycero-3-phosphoethanolamine-N- [methoxy (polyethylene glycol) -3000 ] ) or analog . In an embodiment of any one of the compositions or methods provided herein, the polymer is 16 : 0 PEG3000 PE ( 1 , 2-dipalmitoyl-sn-glycero-3-phosphoethanolamine-N- [methoxy (polyethylene glycol) -3000 ] ) or analog . In an embodiment of any one of the compositions or methods provided herein, the polymer is 18 : 0 PEG3000 PE ( 1 , 2-distearoyl-sn-glycero-3-phosphoethanolamine-N- [methoxy (polyethylene glycol) -3000 ] ) or analog . In an embodiment of any one of the compositions or methods provided herein, the polymer is 18 : 1 PEG3000 PE ( 1 , 2-dioleoyl-sn-glycero-3-phosphoethanolamine-N- [methoxy (polyethylene glycol) -3000 ] ) or analog . In an embodiment of any one of the compositions or methods provided herein, the polymer is a PEG-modified lipid with a polyethylene glycol chain of up to 5 kDa in length . In an embodiment of any one of the compositions or methods provided herein, the polymer is 14 : 0 PEG5000 PE ( 1 , 2-dimyristoyl-sn-glycero-3-phosphoethanolamine-N- [methoxy (polyethylene glycol) -5000 ] ) or analog. In an embodiment of any one of the compositions or methods provided herein, the polymer is 16 : 0 PEG5000 PE ( 1 , 2-dipalmitoyl-sn-glycero-3-phosphoethanolamine-N- [methoxy (polyethylene glycol) -5000 ] ) or analog. In an embodiment of any one of the compositions or methods provided herein, the polymer is 18 : 0 PEG5000 PE ( 1 , 2-distearoyl-sn-glycero-3-phosphoethanolamine-N- [methoxy (polyethylene glycol) -5000 ] ) or analog. In an embodiment of any one of the compositions or methods provided herein, the polymer is DSPE-2arm-PEG5000 (N- [2 ' , 3 ' -Bis (methylpolyoxyethylene oxy) propane-1 ' -oxycarbonyl] -1, 2-distearoyl-sn-glycero-3-phosphoethanolamine) or analog. In an embodiment of any one of the compositions or methods provided herein, the polymer is 18 : 1 PEG5000 PE ( 1 , 2-dioleoyl-sn-glycero-3-phosphoethanolamine-N- [methoxy (polyethylene glycol) -5000 ] ) or analog . In an embodiment of any one of the compositions or methods provided herein, the polymer is a PEG-modified glyceride . In an embodiment of any one of the compositions or methods provided herein, the polymer is DMG-PEG2000 ( 1 , 2-dimyristoyl-rac-glycero-3-methoxypolyethylene glycol-2000 ) or analog . In an embodiment of any one of the compositions or methods provided herein, the polymer is DSG-PEG2000 (a- [ 2 , 3-bis [ ( 1- oxooctadecyl) oxy] propyl] -w-methoxy-poly (oxy-1, 2-ethanediyl) ) or analog . In an embodiment of any one of the compositions or methods provided herein, the polymer is DPG-PEG2000 ( 1 , 2-Dipalmitoyl-rac-glycero-3-methylpolyoxyethylene) or analog . In an embodiment of any one of the compositions or methods provided herein, the polymer is DOG-PEG2000 (1, 2-Dioleoyl-rac-glycerol, methoxypolyethylene Glycol) or analog . In an embodiment of any one of the compositions or methods provided herein, the polymer is DMG-PEG5000 ( 1 , 2-Dimyristoyl-rac-glycero-3-methylpolyoxyethylene) or analog . In an embodiment of any one of the compositions or methods provided herein, the polymer is DSG-PEG5000 (1, 2-Distearoyl-rac-glycero-3-methylpolyoxyethylene) or analog . In an embodiment of any one of the compositions or methods provided herein, the polymer is DPG-PEG5000 ( 1 , 2-Dipalmitoyl-rac-glycerol, methoxypolyethylene Glycol) or analog. In an embodiment of any one of the compositions or methods provided herein, the polymer is DOG-PEG5000 (1, 2-Dioleoyl-rac-glycerol, methoxypolyethylene Glycol) or analog . In an embodiment of any one of the compositions or methods provided herein, the polymer is Sunbright GS-020MA (PEG2000 , 1, 2-Distearoyl-rac-glycero-3- (2' -maleimidoethyl) polyoxyethylene) or analog . In an embodiment of any one of the compositions or methods provided herein, the polymer is Sunbright GS-020TS (PEG2000 , 1, 2-Distearoyl-rac-glycero-3- ( succinimidyloxycarbonyl) polyoxyethylene) or analog . In an embodiment of any one of the compositions or methods provided herein, the polymer is ALC-0159

[0023] (Methoxypoly ethylene glycol oxy (2000 ) -N, N-dit etradecyl acet amide) or analog . In an embodiment of any one of the compositions or methods provided herein, the polymer is a PEG-modified sterol . In an embodiment of any one of the compositions or methods provided herein, the polymer is Chol-PEG600 (Cholesterol- (polyethylene glycol-600 ) ) or analog . In an embodiment of any one of the compositions or methods provided herein, the polymer is Chol-PEG2000 (Cholesterol- (polyethylene glycol-2000) ) or analog. In an embodiment of any one of the compositions or methods provided herein, the polymer is a PEG-modified ceramide . In an embodiment of any one of the compositions or methods provided herein, the polymer is C8 PEG750 ceramide (N- o ct anoyl-sphingo sine-1- { succinyl [methoxy (polyethylene glycol) 750 ] } ) or analog . In an embodiment of any one of the compositions or methods provided herein, the polymer is C16 PEG750 ceramide (N-palmitoyl-sphingosine-1- { succinyl [methoxy (polyethylene glycol) 750 ] } ) or analog . In an embodiment of any one of the compositions or methods provided herein, the polymer is C8 PEG2000 ceramide (N-octanoyl-sphingosine-1- { succinyl [methoxy (polyethylene glycol) 2000 ] } ) or analog . In an embodiment of any one of the compositions or methods provided herein, the polymer is C16 PEG2000 ceramide (N-palmitoyl-sphingosine-1- { succinyl [methoxy (polyethylene glycol) 2000 ] } ) or analog . In an embodiment of any one of the compositions or methods provided herein, the polymer is C8 PEG5000 ceramide (N-octanoyl-sphingosine-1- { succinyl [methoxy (polyethylene glycol) 5000 ] } ) or analog . In an embodiment of any one of the compositions or methods provided herein, the polymer is C16 PEG5000 ceramide (N-palmitoyl-sphingosine-1- { succinyl [methoxy (polyethylene glycol) 5000 ] } ) or analog . In an embodiment of any one of the compositions or methods provided herein, the polymer is a diacetylene lipid. In an embodiment of any one of the compositions or methods provided herein, the polymer is 23 : 2 Diyne PE ( 1, 2-bis (10, 12-tricosadiynoyl) -sn-glycero-3-phosphoethanolamine) or analog . In an embodiment of any one of the compositions or methods provided herein, the polymer is 16 : 0-23 : 2 Diyne PE (PTPE, 1-palmitoyl-2- (10, 12-tricosadiynoyl) -sn-glycero-3-phosphoethanolamine) or analog . In an embodiment of any one of the compositions or methods provided herein, the polymer is 23 : 2 Diyne PC ( 1, 2-bis ( 10 , 12-tricosadiynoyl) -sn-glycero-3-phosphocholine) or analog . In an embodiment of any one of the compositions or methods provided herein, the polymer is 16 : 0-23 : 2 Diyne PC (PTPC, l-palmitoyl-2- (10, 12-tricosadiynoyl) -sn-glycero-3-phosphocholine) or analog. In an embodiment of any one of the compositions or methods provided herein, the polymer is Tocofersolan (TPGS 1000 , D-a-tocopheryl polyethylene glycol succinate) or analog .

[0024] In an embodiment of any one of the compositions or methods provided herein, the intracellular vesicle-interacting molecule is a cellpenetrating molecule . In an embodiment of any one of the compositions or methods provided herein, the intracellular vesicle-interacting molecule is CPM2 or analog . In an embodiment of any one of the compositions or methods provided herein, the intracellular vesicleinteracting molecule is a peptide .

[0025] In an embodiment of any one of the compositions or methods provided herein, the intracellular vesicle-interacting peptide is a membrane-disruptive peptide . In an embodiment of any one of the compositions or methods provided herein, the intracellular vesicle-interacting peptide is aurein 1.2 (GLFDIIKKIAESF) or analog. In an embodiment of any one of the compositions or methods provided herein, the intracellular vesicle-interacting peptide is a cell-penetrating peptide . In an embodiment of any one of the compositions or methods provided herein, the intracellular vesicle-interacting peptide is an endosomedisrupting peptide . In an embodiment of any one of the compositions or methods provided herein, the intracellular vesicle-interacting peptide is a truncated form of the N-terminus of adenovirus protein VI (LLLLAFSWGSLWSGIKNFGSTIKN) or analog . In an embodiment of any one of the compositions or methods provided herein, the intracellular vesicle-interacting peptide is Tat ( YGRKKRRQRRR) or analog. In an embodiment of any one of the compositions or methods provided herein, the intracellular vesicle-interacting peptide is dTat-HA2 (D-Tat fused to HA2 ) or analog. In an embodiment of any one of the compositions or methods provided herein, the intracellular vesicle-interacting peptide is transportan 10 (AGYLLGKINLKALAALAKKIL) or analog. In an embodiment of any one of the compositions or methods provided herein, the intracellular vesicle-interacting peptide is octaarginine (R8 ) or analog . In an embodiment of any one of the compositions or methods provided herein, the intracellular vesicle-interacting peptide is nonaarginine (R9) or analog. In an embodiment of any one of the compositions or methods provided herein, the intracellular vesicleinteracting peptide is octalysine (L8) or analog . In an embodiment of any one of the compositions or methods provided herein, the intracellular vesicle-interacting peptide is penetratin (RQIKIWFQNRRMKWKK) or analog . In an embodiment of any one of the compositions or methods provided herein, the intracellular vesicleinteracting peptide is a histidine-switching cell-penetrating peptide . In an embodiment of any one of the compositions or methods provided herein, the intracellular vesicle-interacting peptide is hsTAT

[0026] ( YGHKKHHQHHH) or analog. In an embodiment of any one of the compositions or methods provided herein, the intracellular vesicleinteracting peptide is hsTPIO (FSGSGSGSGAGYLLGHINLHALAALAHHILGC) or analog . In an embodiment of any one of the compositions or methods provided herein, the intracellular vesicle-interacting peptide is hsPen (FSGSGSGSGHQIHIWFQNHHMHWHHGC) or analog. In an embodiment of any one of the compositions or methods provided herein, the intracellular vesicle-interacting peptide is hsTPT (CLIHHALAALAHLNIHLLYGASNLTWG) or analog . In an embodiment of any one of the compositions or methods provided herein, the intracellular vesicle-interacting peptide is hsLMWP (GSVSHHHHHHGGHHHH) or analog. In an embodiment of any one of the compositions or methods provided herein, the intracellular vesicle-interacting peptide is hsR12 (HHHHHHHHHHHH) or analog . In an embodiment of any one of the compositions or methods provided herein, the intracellular vesicle-interacting peptide is hsMPG (GALFLGFLGAAGSTMGAWSQPHHHHHV) or analog. In an embodiment of any one of the compositions or methods provided herein, the intracellular vesicle-interacting peptide is a fusogenic peptide . In an embodiment of any one of the compositions or methods provided herein, the intracellular vesicle-interacting peptide is CM18 or analog . In an embodiment of any one of the compositions or methods provided herein, the intracellular vesicle-interacting peptide is CM18-Tat (KWKLFKKIGAVLKVLTTG-YGRKKRRQRRR) or analog . In an embodiment of any one of the compositions or methods provided herein, the intracellular vesicle-interacting peptide is diINF-7 or analog . In an embodiment of any one of the compositions or methods provided herein, the intracellular vesicle-interacting peptide is H5WYG or analog. In an embodiment of any one of the compositions or methods provided herein, the intracellular vesicle-interacting peptide is a pH-sensitive fusogenic peptide . In an embodiment of any one of the compositions or methods provided herein, the intracellular vesicle-interacting peptide is HA2 (GLFGAIAGFIENGWEGMIDGWYG) or analog . In an embodiment of any one of the compositions or methods provided herein, the intracellular vesicle-interacting peptide is GALA (WEAALAEALAEALAEHLAEALAEALEALAA) or analog. In an embodiment of any one of the compositions or methods provided herein, the intracellular vesicle-interacting peptide an amphiphilic peptide . In an embodiment of any one of the compositions or methods provided herein, the intracellular vesicle-interacting peptide is 6His-CM18-PTD4 or analog. In an embodiment of any one of the compositions or methods provided herein, the intracellular vesicle-interacting peptide is pHlip or analog . In an embodiment of any one of the compositions or methods provided herein, the intracellular vesicle-interacting peptide is pHD118 or analog . In an embodiment of any one of the compositions or methods provided herein, the intracellular vesicle-interacting peptide is a cyclic cellpenetrating peptide . In an embodiment of any one of the compositions or methods provided herein, the intracellular vesicle-interacting peptide is cF4>R4 (cyclic phenylalanine-L-2-naphthylalanine-tetraarginine) or analog . In an embodiment of any one of the compositions or methods provided herein, the intracellular vesicleinteracting peptide is CPP9 or analog. In an embodiment of any one of the compositions or methods provided herein, the intracellular vesicle-interacting peptide is CPP12 or analog . In an embodiment of any one of the compositions or methods provided herein, the intracellular vesicle-interacting peptide is palmitoyl-cyclic-dodeca-D-arginine (palmitoyl-GSSGCRRRRRRRRRRRRC-NH2 ) or analog . In an embodiment of any one of the compositions or methods provided herein, the intracellular vesicle-interacting peptide is a miniature protein . In an embodiment of any one of the compositions or methods provided herein, the intracellular vesicle-interacting peptide is miniature protein 5.3. In an embodiment of any one of the compositions or methods provided herein, the intracellular vesicle-interacting peptide is miniature protein ZF5.3. In an embodiment of any one of the compositions or methods provided herein, the intracellular vesicleinteracting peptide contains an endosomal escape domain . In an embodiment of any one of the compositions or methods provided herein,

[0027] 1 the intracellular vesicle-interacting peptide contains the penetration accelerating sequence GKPILFF .

[0028] In an embodiment of any one of the compositions or methods provided herein, the cholesterol or cholesterol analog is cholesterol ( ( 313 ) — cholest-5-en-3-ol) or analog . In an embodiment of any one of the compositions or methods provided herein, the cholesterol or cholesterol analog is desmosterol (3f>-hydroxy-5, 24-cholestadiene) or analog. In an embodiment of any one of the compositions or methods provided herein, the cholesterol or cholesterol analog is lanosterol (lanosta-8, 24-dien-3p-ol) or analog. In an embodiment of any one of the compositions or methods provided herein, the cholesterol or cholesterol analog is lanosterol-95 (8 , 24-lanostadien-3b-ol) or analog. In an embodiment of any one of the compositions or methods provided herein, the cholesterol or cholesterol analog is 14-demethyl-lanosterol ( 4 , 4-dimethyl cholest-8 ( 9) , 24-dien-3f>-ol) or analog. In an embodiment of any one of the compositions or methods provided herein, the cholesterol or cholesterol analog is zymosterol ( 5a-cholesta-8, 24-dien-3f>-ol) or analog. In an embodiment of any one of the compositions or methods provided herein, the cholesterol or cholesterol analog is lathosterol ( 5a-cholest-7-en-3f>-ol) or analog. In an embodiment of any one of the compositions or methods provided herein, the cholesterol or cholesterol analog is 24-methylene cholesterol ( 5, 24 (28 ) -cholestadien-24-methylen-3f>-ol) or analog . In an embodiment of any one of the compositions or methods provided herein, the cholesterol or cholesterol analog is a dehydrocholesterol . In an embodiment of any one of the compositions or methods provided herein, the cholesterol or cholesterol analog is 7-dehydrocholesterol (A5, 7-cholesterol) or analog. In an embodiment of any one of the compositions or methods provided herein, the cholesterol or cholesterol analog is 8 (9) — dehydrocholesterol (cholesta-5, 8 ( 9) -dien-3f>-ol) or analog . In an embodiment of any one of the compositions or methods provided herein, the cholesterol or cholesterol analog is 8 (14) -dehydrocholesterol (cholesta-5, 8 (14) -dien-3f>-ol) or analog. In an embodiment of any one of the compositions or methods provided herein, the cholesterol or cholesterol analog is FF-MAS ( 14-demethyl-14-dehydrolanosterol) or analog . In an embodiment of any one of the compositions or methods provided herein, the cholesterol or cholesterol analog is a saturated sterol . In an embodiment of any one of the compositions or methods provided herein, the cholesterol or cholesterol analog is dihydrolanosterol (24, 25-dihydrolanosterol) or analog . In an embodiment of any one of the compositions or methods provided herein, the cholesterol or cholesterol analog is zymostenol ( 5 e -Cholest-8-en-3-beta-ol) or analog. In an embodiment of any one of the compositions or methods provided herein, the cholesterol or cholesterol analog is a cholesterol ester . In an embodiment of any one of the compositions or methods provided herein, the cholesterol or cholesterol analog is cholesteryl nervonate (cholest-5-en-3-ol, 3p- [ (15Z) -15-tetracosenoate] ) or analog . In an embodiment of any one of the compositions or methods provided herein, the cholesterol or cholesterol analog is cholesteryl oleate (cholest-5-en-3p-ol, 3— [ ( 9Z) — 9-octadecenoate ] ) or analog. In an embodiment of any one of the compositions or methods provided herein, the cholesterol or cholesterol analog is cholesteryl palmitate ( (3p) -cholest-5-en-3-ol, 3-hexadecanoate) or analog . In an embodiment of any one of the compositions or methods provided herein, the cholesterol or cholesterol analog is 17 : 0 cholesteryl ester ( cholest-5-en-3f>-yl heptadecanoate) or analog. In an embodiment of any one of the compositions or methods provided herein, the cholesterol or cholesterol analog is cholesteryl stearate ( (3p) -cholest-5-en-3-ol, 3-octadecanoate) or analog . In an embodiment of any one of the compositions or methods provided herein, the cholesterol or cholesterol analog is cholesteryl lignocerate (cholest-5-en- (3p) -ol, 3-tetracosanoate) or analog . In an embodiment of any one of the compositions or methods provided herein, the cholesterol or cholesterol analog is cholesteryl homo-y_linolenate ( cholest-5-en-3 p-ol, 3- ( 8Z, 11Z, 14Z) -8 , 11, 14-eicosatrienoate) or analog . In an embodiment of any one of the compositions or methods provided herein, the cholesterol or cholesterol analog is an oxysterol . In an embodiment of any one of the compositions or methods provided herein, the cholesterol or cholesterol analog is 4p-hydroxy cholesterol (cholest-5-ene-3p, 4 p-diol) or analog . In an embodiment of any one of the compositions or methods provided herein, the cholesterol or cholesterol analog is 25-hydroxy cholesterol (cholest-5-ene-3p, 25-diol) or analog. In an embodiment of any one of the compositions or methods provided herein, the cholesterol or cholesterol analog is a phytosterol (derived from plants) . In an embodiment of any one of the compositions or methods provided herein, the cholesterol or cholesterol analog is stigmasterol ( stigmasta-5, 22E-dien-3p-ol) or analog. In an embodiment of any one of the compositions or methods provided herein, the cholesterol or cholesterol analog is 24-methyl cholesterol ( (3p, 24 -ergost-5-en-3-ol) or analog. In an embodiment of any one of the compositions or methods provided herein, the cholesterol or cholesterol analog is p-Sitosterol ( stigmast-5-en-3p-ol) or analog . In an embodiment of any one of the compositions or methods provided herein, the cholesterol or cholesterol analog is brassicasterol (ergosta-5, 22E-dien-3p-ol) or analog . In an embodiment of any one of the compositions or methods provided herein, the cholesterol or cholesterol analog is fucosterol ( (3p, 24E) -stigmasta-5, 24 (28 ) -dien-3-ol) or analog. In an embodiment of any one of the compositions or methods provided herein, the cholesterol or cholesterol analog is a-spinasterol ( (3p, 5a, 22E) -stigmasta-7, 22-dien-3-ol) or analog . In an embodiment of any one of the compositions or methods provided herein, the cholesterol or cholesterol analog is campesterol ( campest-5-en-3f>-ol) or analog . In an embodiment of any one of the compositions or methods provided herein, the cholesterol or cholesterol analog is a saturated phytosterol . In an embodiment of any one of the compositions or methods provided herein, the cholesterol or cholesterol analog is sitostanol ( (5a) -stigmastan-3p-ol) or analog. In an embodiment of any one of the compositions or methods provided herein, the cholesterol or cholesterol analog is campestanol

[0029] ( (3p, 5a, 24R) -ergostan-3-ol) or analog.

[0030] In an embodiment of any one of the compositions or methods provided herein, the cholesterol substitute is a sterol or sterol derivative . In an embodiment of any one of the compositions or methods provided herein, the cholesterol substitute is a chemical or biological substance that can modify lipid membrane fluidity or rigidity over a range of physiological temperatures . In an embodiment of any one of the compositions or methods provided herein, the cholesterol substitute contains at least one polar group such as a hydroxyl, at least one segment comprised of at least 3 aliphatic rings, and at least one hydrophobic group comprised of at least 3 and at most 16 carbon atoms .

[0031] In an embodiment of any one of the compositions or methods provided herein, the targeting element is an antibody . In an embodiment of any one of the compositions or methods provided herein, the targeting element is an antibody derivative such as a nanobody, antibody fragment, bispecific, antibody variable region or a combination of one or more antibody derivatives . In an embodiment of any one of the compositions or methods provided herein, the targeting element is anti-c-Kit (anti-CD117 ) . In an embodiment of any one of the compositions or methods provided herein, the targeting element is anti-glucocorticoid receptor . In an embodiment of any one of the compositions or methods provided herein, the targeting element is anti-transforming growth factor beta receptor 2 (TGFBR2 ) . In an embodiment of any one of the compositions or methods provided herein, the targeting element is anti-transforming growth factor beta receptor 3 (TGFBR3) . In an embodiment of any one of the compositions or methods provided herein, the targeting element is anti-aquaporin 7. In an embodiment of any one of the compositions or methods provided herein, the targeting element is anti-aquaporin 7B . In an embodiment of any one of the compositions or methods provided herein, the targeting element is anti-activin A receptor type 1C . In an embodiment of any one of the compositions or methods provided herein, the targeting element is anti-frizzled class 4 receptor . In an embodiment of any one of the compositions or methods provided herein, the targeting element is anti-perilipin 1. In an embodiment of any one of the compositions or methods provided herein, the targeting element is anti-perilipin 4. In an embodiment of any one of the compositions or methods provided herein, the targeting element is anti-SLC7A10 . In an embodiment of any one of the compositions or methods provided herein, the targeting element is anti-platelet-derived growth factor receptor alpha . In an embodiment of any one of the compositions or methods provided herein, the targeting element is anti-platelet-derived growth factor receptor beta . In an embodiment of any one of the compositions or methods provided herein, the targeting element is a nanobody. In an embodiment of any one of the compositions or methods provided herein, the targeting element is an antigen-binding fragment (Fab) . In an embodiment of any one of the compositions or methods provided herein, the targeting element is a bispecific or multi-specific antibody or antibody fragment . In an embodiment of any one of the compositions or methods provided herein, the targeting element is comprised at least in part of an antibody variable region . In an embodiment of any one of the compositions or methods provided herein, the targeting element is a protein . In an embodiment of any one of the compositions or methods provided herein, the targeting element is angiotensin II . In an embodiment of any one of the compositions or methods provided herein, the targeting element is VEGF . In an embodiment of any one of the compositions or methods provided herein, the targeting element is wheat germ agglutinin (WGA) . In an embodiment of any one of the compositions or methods provided herein, the targeting element is Ulex europaeus agglutinin I (UEA-1) . In an embodiment of any one of the compositions or methods provided herein, the targeting element is a peptide . In an embodiment of any one of the compositions or methods provided herein, the targeting element is miniAp-4 . In an embodiment of any one of the compositions or methods provided herein, the targeting element is PEP3. In an embodiment of any one of the compositions or methods provided herein, the targeting element is a peptide with the sequence CKGGRAKDC . In an embodiment of any one of the compositions or methods provided herein, the targeting element is a peptide with the sequence GGGGYDRVTIHPF . In an embodiment of any one of the compositions or methods provided herein, the targeting element is a peptide with the sequence D (KLAKLAK) 2. In an embodiment of any one of the compositions or methods provided herein, the targeting element is a peptide with the sequence MRYMILGLLALAAVCSA . In an embodiment of any one of the compositions or methods provided herein, the targeting element is is a receptor ligand. In an embodiment of any one of the compositions or methods provided herein, the targeting element is LJ-4378 (agonist of A2A adenosine receptor / antagonist of A3 adenosine receptor) . In an embodiment of any one of the compositions or methods provided herein, the targeting element is resistin (ligand of decorin lacking the glycanation site) . In an embodiment of any one of the compositions or methods provided herein, the targeting element is norrin ( specific ligand of frizzled class 4 receptor) . In an embodiment of any one of the compositions or methods provided herein, the targeting element is a receptor binding domain . In an embodiment of any one of the compositions or methods provided herein, the targeting element is a lipid. In an embodiment of any one of the compositions or methods provided herein, the targeting element is PAHSA (palmitic-acid-9-hydroxy-stearic-acid) or analog . In an embodiment of any one of the compositions or methods provided herein, the targeting element is a polysaccharide . In an embodiment of any one of the compositions or methods provided herein, the targeting element is glycogen . In an embodiment of any one of the compositions or methods provided herein, the targeting element is glucan . In an embodiment of any one of the compositions or methods provided herein, the targeting element is dextran . In an embodiment of any one of the compositions or methods provided herein, the targeting element is chitosan . In an embodiment of any one of the compositions or methods provided herein, the targeting element is alginate . In an embodiment of any one of the compositions or methods provided herein, the targeting element is hyaluronic acid. In an embodiment of any one of the compositions or methods provided herein, the targeting element is a nucleic acid. In an embodiment of any one of the compositions or methods provided herein, the targeting element is an aptamer . In an embodiment of any one of the compositions or methods provided herein, the targeting element is an aptamer against VEGFR. In an embodiment of any one of the compositions or methods provided herein, the nucleic acid is a double-stranded deoxyribonucleic acid (DNA) . In an embodiment of any one of the compositions or methods provided herein, the nucleic acid is a single-stranded DNA. In an embodiment of any one of the compositions or methods provided herein, the nucleic acid is a linear DNA. In an embodiment of any one of the compositions or methods provided herein, the nucleic acid is a circular DNA. In an embodiment of any one of the compositions or methods provided herein, the nucleic acid is a DNA 0. 5 to 50 kb in length . In an embodiment of any one of the compositions or methods provided herein, the nucleic acid is a DNA 1.5 to 7.0 kb in length .

[0032] In an embodiment of any one of the compositions or methods provided herein, the genetic cargo encodes at least a promoter and / or enhancer . In an embodiment of any one of the compositions or methods provided herein, the genetic cargo encodes at least the cytomegalovirus (CMV) immediate early promoter . In an embodiment of any one of the compositions or methods provided herein, the genetic cargo encodes at least the eukaryotic translation elongation factor 1 al (EF1A) promoter . In an embodiment of any one of the compositions or methods provided herein, the genetic cargo encodes at least the eukaryotic translation elongation factor 1 al short form (EFS) promoter . In an embodiment of any one of the compositions or methods provided herein, the genetic cargo encodes at least the CAG promoter . In an embodiment of any one of the compositions or methods provided herein, the genetic cargo encodes at least the CBh promoter . In an embodiment of any one of the compositions or methods provided herein, the genetic cargo encodes at least the CBA promoter . In an embodiment of any one of the compositions or methods provided herein, the genetic cargo encodes at least the simian virus 40 (SV40) enhancer / early promoter . In an embodiment of any one of the compositions or methods provided herein, the genetic cargo encodes at least the phosphoglycerate kinase 1 (BGK) promoter . In an embodiment of any one of the compositions or methods provided herein, the genetic cargo encodes at least an adiposespecific promoter . In an embodiment of any one of the compositions or methods provided herein, the genetic cargo encodes at least the adipocyte P2 (aP2) promoter . In an embodiment of any one of the compositions or methods provided herein, the genetic cargo encodes at least the adiponectin promoter . In an embodiment of any one of the compositions or methods provided herein, the genetic cargo encodes at least the peroxisome prolif erator-activated receptor gamma (PPAR gamma) promoter . In an embodiment of any one of the compositions or methods provided herein, the genetic cargo encodes at least the fatty acid binding protein 4 (FABP4) promoter . In an embodiment of any one of the compositions or methods provided herein, the genetic cargo encodes at least the laeverin (LVRN) promoter . In an embodiment of any one of the compositions or methods provided herein, the genetic cargo encodes at least the perilipin 1 (PLIN1) promoter . In an embodiment of any one of the compositions or methods provided herein, the genetic cargo encodes at least the perilipin 4 (PLIN4) promoter . In an embodiment of any one of the compositions or methods provided herein, the genetic cargo encodes at least a therapeutic transgene . In an embodiment of any one of the compositions or methods provided herein, the genetic cargo encodes at least insulin . In an embodiment of any one of the compositions or methods provided herein, the genetic cargo encodes at least gastric inhibitory peptide (GIP) receptor agonist or analog . In an embodiment of any one of the compositions or methods provided herein, the genetic cargo encodes at least glucagon-like peptide 1 (GLP1) receptor agonist or analog. In an embodiment of any one of the compositions or methods provided herein, the genetic cargo encodes at least a GIP / GLP1 receptor co-agonist or analog . In an embodiment of any one of the compositions or methods provided herein, the genetic cargo encodes at least insulin-like growth factor 1 ( IGF1) . In an embodiment of any one of the compositions or methods provided herein, the genetic cargo encodes at least insulin-like growth factor 2 (IGF2) . In an embodiment of any one of the compositions or methods provided herein, the genetic cargo encodes at least beta-glucocerebrosidase . In an embodiment of any one of the compositions or methods provided herein, the genetic cargo encodes at least an alpha-glucosidase . In an embodiment of any one of the compositions or methods provided herein, the genetic cargo encodes at least fibroblast growth factor 18 (FGF18 ) . In an embodiment of any one of the compositions or methods provided herein, the genetic cargo encodes at least vascular endothelial growth factor (VEGFA) . In an embodiment of any one of the compositions or methods provided herein, the genetic cargo encodes at least factor VIII . In an embodiment of any one of the compositions or methods provided herein, the genetic cargo encodes at least factor IX . In an embodiment of any one of the compositions or methods provided herein, the genetic cargo encodes at least liver-expressed antimicrobial peptide 2 (LEAP2,

[0033] MTPFWRGVSLRP IGASCRDDSECITRLCRKRRCSLSVAQE ) . In an embodiment of any one of the compositions or methods provided herein, the genetic cargo encodes at least an siRNA. In an embodiment of any one of the compositions or methods provided herein, the genetic cargo encodes at least an siRNA against X-linked inhibitor of apoptosis-associated factor 1 (XAF1) . In an embodiment of any one of the compositions or methods provided herein, the genetic cargo encodes at least an shRNA. In an embodiment of any one of the compositions or methods provided herein, the genetic cargo encodes at least an antibody. In an embodiment of any one of the compositions or methods provided herein, the genetic cargo encodes at least an antibody fragment . In an embodiment of any one of the compositions or methods provided herein, the genetic cargo encodes at least a nanobody. In an embodiment of any one of the compositions or methods provided herein, the genetic cargo encodes at least an anti-tumor necrosis factor alpha (TNF-alpha) nanobody . In an embodiment of any one of the compositions or methods provided herein, the genetic cargo encodes at least a post-transcriptional regulatory element . In an embodiment of any one of the compositions or methods provided herein, the genetic cargo encodes at least the hepatitis B post-transcriptional regulatory element (HPRE) . In an embodiment of any one of the compositions or methods provided herein, the genetic cargo encodes at least the Woodchuck Hepatitis Virus post-transcriptional regulatory element (WPRE) . In an embodiment of any one of the compositions or methods provided herein, the genetic cargo encodes at least the optimized Woodchuck Hepatitis Virus post- transcriptional regulatory element (oPRE) . In an embodiment of any one of the compositions or methods provided herein, the genetic cargo encodes at least polyadenylation sequence . In an embodiment of any one of the compositions or methods provided herein, the genetic cargo encodes at least the bovine growth hormone polyadenylation sequence (bGH pA) . In an embodiment of any one of the compositions or methods provided herein, the genetic cargo encodes at least the SV40 polyadenylation sequence (SV40 pA) . In an embodiment of any one of the compositions or methods provided herein, the genetic cargo encodes at least a downtitration element . In an embodiment of any one of the compositions or methods provided herein, the genetic cargo encodes at least a suicide gene . In an embodiment of any one of the compositions or methods provided herein, the genetic cargo encodes iCasp8 (inducible caspase 8 ) . In an embodiment of any one of the compositions or methods provided herein, the genetic cargo encodes iCasp9 (inducible caspase 9) .

[0034] In an embodiment of any one of the compositions or methods provided herein, the nanoparticle formulation has an average diameter of 25 -250 nm. In an embodiment of any one of the compositions or methods provided herein, the nanoparticle formulation has an average diameter of 35 - 150 nm. In an embodiment of any one of the compositions or methods provided herein, the nanoparticle formulation has a zeta potential in the range of -0 .5 to -15. 0 mV. In an embodiment of any one of the compositions or methods provided herein, the nanoparticle formulation has a zeta potential in the range of -6.0 to -10. 0 mV. In an embodiment of any one of the compositions or methods provided herein, the nanoparticle formulation has a polydispersity index of 0 . 001 to 2 .50. In an embodiment of any one of the compositions or methods provided herein, the nanoparticle formulation has a polydispersity index of 0. 01 to 0.3. In an embodiment of any one of the compositions or methods provided herein, the nanoparticle formulation has an N to P ratio of 1.5 to 20 . In an embodiment of any one of the compositions or methods provided herein, the nanoparticle formulation has an N to P ratio of 3.5 to 9. 5. In an embodiment of any one of the compositions or methods provided herein, the nanoparticle formulation comprises ionizable lipid ( s) comprising a molar ratio of about 15-65% . In an embodiment of any one of the compositions or methods provided herein, the nanoparticle formulation comprises structural element (s) comprising molar ratio of about 0-35% of the total lipid present in the formulation . In an embodiment of any one of the compositions or methods provided herein, the nanoparticle formulation comprises polymer ( s) comprising a molar ratio of about 0-20% of the total lipid present in the formulation . In an embodiment of any one of the compositions or methods provided herein, the nanoparticle formulation comprises sterol ( s) , cholesterol ( s ) , or cholesterol-like molecule ( s) comprising a molar ratio of about 0-55% of the total lipid present in the formulation .

[0035] In an embodiment of any one of the compositions or methods provided herein, the nanoparticle formulation is for treating diabetes . In an embodiment of any one of the compositions or methods provided herein, the nanoparticle formulation is for treating treating growth restriction . In an embodiment of any one of the compositions or methods provided herein, the nanoparticle formulation is for treating rheumatoid arthritis . In an embodiment of any one of the compositions or methods provided herein, the nanoparticle formulation is for treating Gaucher' s disease . In an embodiment of any one of the compositions or methods provided herein, the nanoparticle formulation is for treating progeria . In an embodiment of any one of the compositions or methods provided herein, the nanoparticle formulation is for treating Pompe disease . In an embodiment of any one of the compositions or methods provided herein, the nanoparticle formulation is for treating hemophilia .

[0036] In an aspect is a method comprising administering any one of the compositions provided herein to a subject in need thereof, such as any one of the conditions or diseases or disorders provided herein .

[0037] BRIEF DESCRIPTION OF THE FIGURES FIG. 1 demonstrates transfection efficiency as described in Example 1. FIG. 1A demonstrates transfection efficiency as a function of N / P ratio with primary human adipocytes at 72h . FIG. IB demonstrates transfection efficiency as a function of N / P ratio with primary human adipocytes at 120h .

[0038] FIG. 2 shows the % GFP positive cells (FIG. 2A in adipocytes and FIG.

[0039] 2B in preadipocytes) and cell count / nuclear stain (FIG. 2C in adipocytes and FIG. 2D in preadipocytes) as described in Example 1.

[0040] FIG. 3 shows the % GFP positive cells (FIG. 3A in adipocytes and FIG.

[0041] 3B in preadipocytes) and cell count / nuclear stain (FIG. 3C in adipocytes and FIG. 3D in preadipocytes) as described in Example 1.

[0042] FIG. 4 demonstrates statistically significant weight loss (FIG. 4A) , improved insulin response on ITT (FIG. 4B) and improved glucose response on GTT (FIG. 4C) as described in Example 1.

[0043] FIG. 5 shows staining as described in Example 6.

[0044] FIG. 6 shows staining as described in Example 6.

[0045] FIG. 7 shows staining as described in Example 6.

[0046] FIG. 8 shows staining as described in Example 6 (FIG. 8A) and imaging (FIG. 8B) as described in Example 6.

[0047] FIG. 9 provides results with adipocyte targeted genetic constructs verus background (FIG. 9A) and with adipocyte targeted vector compositions versus background (FIG. 9B) as described in Example 6.

[0048] FIG. 10 shows signal biodistribution at necropsy in a number of organs (FIG. 10A) and imaging (FIG. 10B) as described in Example 6. DETAILED DESCRIPTION OF THE INVENTION

[0049] Before describing the present invention in detail, it is to be understood that this invention is not limited to particularly exemplified materials or process parameters as such may, of course, vary . It is also to be understood that the terminology used herein is for the purpose of describing particular embodiments of the invention only, and is not intended to be limiting of the use of alternative terminology to describe the present invention .

[0050] All publications, patents and patent applications cited herein, whether supra or infra, are hereby incorporated by reference in their entirety for all purposes . Such incorporation by reference is not intended to be an admission that any of the incorporated publications, patents and patent applications cited herein constitute prior art .

[0051] As used in this specification and the appended claims, the singular forms "a, " "an" and "the" include plural referents unless the content clearly dictates otherwise . For example, reference to "a molecule" includes a mixture of two or more such molecules or a plurality of such molecules, and the like .

[0052] As used herein, the term "comprise" or variations thereof such as "comprises" or "comprising" are to be read to indicate the inclusion of any recited integer (e . g. a feature, element, characteristic, property, method / process step or limitation) or group of integers (e . g . features, elements, characteristics, properties, method / process steps or limitations) but not the exclusion of any other integer or group of integers . Thus, as used herein, the term "comprising" is inclusive and does not exclude additional, unrecited integers or method / process steps . In embodiments of any of the compositions and methods provided herein, "comprising" may be replaced with "consisting essentially of" or "consisting of" . The phrase "consisting essentially of" is used herein to require the specified integer (s) or steps as well as those which do not materially affect the character or function of the claimed invention . As used herein, the term "consisting" is used to indicate the presence of the recited integer (e . g. a feature, element, characteristic, property, method / process step or limitation) or group of integers (e . g. features, elements, characteristics, properties, method / process steps or limitations) alone .

[0053] The present invention generally describes, at least in part, nanoparticle formulations comprised of a combination of lipids and / or polymers, and optionally intracellular vesicle interacting peptides, as well as genetic elements for delivery of therapeutic transgenes into adipose tissue or adipose tissues .

[0054] In general, the present invention describes, at least in part, a nanoparticle formulation for the delivery of genetic cargo to subcutaneous adipose tissues, which is comprised of lipids and / or polymers, with or without targeting peptides, with or without intracellular vesicle-interacting peptides or ionizable elements . An intracellular vesicle may be an endosome, caveosome, macropinosome, phagosome, or other intracellular vesicle . The nanoparticle formulation may range from 25nm to 250nm in average diameter, such as 35 to 150nm in average diameter . The nanoparticle formulation may have a zeta potential within the range of -0 . 5mV to -15. 0mV, such as within the range of -6. 0mV to -lO . OmV. The nanoparticle formulation may have a broadly ranging polydispersity, such as 0 .001 to 2.50 PDI and 0 .01 to 0 .3 PDI . The nanoparticle formulations described herein may be formulated at an N to P ratio of 1.5 to 20, such as formulated at an N to P ratio of 3. 5 to 9.5. As used herein, the N to P ratio is defined as the molar ratio of nitrogen in the ionizable lipid to phosphorus in the DNA cargo . The nanoparticle formulation generally is intended to contain DNA cargo, such as of circular or linear DNA, single stranded or double stranded, with or without primers and with or without auxiliary peptides or proteins, but generally ranging from 0. 5 to 50kb in length, such as between 1. 5 and 7.0kb in length . The nanoparticle formulation may be targeted to tissues, organs, or cell types via size, zeta potential, lipid composition or via one or more other means, or may optionally be conjugated or coupled with polypeptide or protein targeting ligands . The compositions provided herein may contain an ionizable lipid, ionizable polymer, cationic lipid, cationic polymer, cholesterol or cholesterol analog, structural lipid or lipidoid, or a lipid-like molecule, or a combination of one or more aforementioned elements with a pKa of about 5 to 8, such as about 6 to 7. As used herein, an ionizable lipid or ionizable polymer is defined as a lipid or polymer that changes its charge to increase its overall cationic charge following acidification of an endosome or generally a change in pH of an intracellular vesicle . The formulation may be produced by impingement mixing, microfluidic mixing, homogenization, or any other method of producing the nanoparticle formulation with the goal of achieving encapsulation efficiency of above 20%, such as above 60%, and such as above 75% prior to purification . The compositions provided herein may have other elements that change the rigidity or structural integrity of the nanoparticle, means of its interaction with the extracellular space, cell surface, or intracellular compartments, or structural elements that include surface modifying peptides or chemical, nanoparticle membrane modifying chemical or biological agents, or biological or chemical agents that interact with cellular components to facilitate endocytosis, uptake, endosomal escape, or translocation of the nanoparticle to one or more specific cellular compartments .

[0055] Generally, provided herein, at least in part, are lipid, polymer, or lipid-polymer nanoparticle formulations for the delivery to a broad range of tissues, such as adipose tissue or another non-life sustaining tissue, and that may include an ionizable lipid (or lipidoid, lipomer, or lipid-like molecule) , a structural lipid, a cationic lipid, a cholesterol or cholesterol analog, and a genetic element . As used herein, "lipid nanoparticle" can be a particle made of multiple lipids that are associated with each other through molecular forces . These may be liposomes, lipoplexes, stabilized plasmid-lipid particles (SPLPs) , stable nucleic acid lipid particles (SNALPs) , micelles, inverted micelles, solid lipid nanoparticles (SLNs) , nanostructured lipid carriers (NLCs) , or an emulsion . As used herein, an ionizable lipid can be a lipid, lipidoid, lipomer, or lipid-like molecule with a transient charge depending on the pH of the surrounding environment . A cationic lipid may be a lipid, lipidoid, lipomer, or lipid-like molecule with a positive charge . Additionally, a structural lipid may be a glycerophospholipid, a neutral lipid, an anionic lipid, or a PEGylated lipid used to distribute or modify the charge of the surface of the lipid nanoparticle, or used to conjugate targeting peptides to the lipid nanoparticle . As used herein, a lipid is considered a molecule with a hydrophilic head group and one or more hydrophobic tails or chains, whose tails may be saturated (with hydrogen) or unsaturated (with double or triple bonds) , or both, and whose tails may be linear, or branched, or both . A cholesterol may be a cholesterol or cholesterol analog, or other sterol lipid. Finally, the genetic element may be a nucleic acid sequence, including a promoter, enhancer, gene, linker sequence, insulators, DNA, or generally genetic sequences, or a combination thereof . The sequence of the genetic element may be optionally codon-optimized for the species of interest .

[0056] In some embodiments, a molecule that is a cholesterol substitute or derivative can be used instead or together with cholesterol or other sterols . Such formulations may therefore contain one or more sterols or sterol derivatives, chemical or biological substances that can modify lipid membrane fluidity or rigidity over a range of physiological temperatures, chemical or biological substances which contain at least one polar group such as a hydroxyl, at least one segment comprised of at least 3 aliphatic rings, and at least one hydrophobic group comprised of at least 3 and at most 16 carbon atoms, or combination of one or more of the aforementioned elements or their derivatives .

[0057] The lipid nanoparticle formulations provided herein may be adjusted to target a specific cell, tissue, or organ or combination thereof . In general, the role of the ionizable lipid is to aid in fusion of the lipid nanoparticle with the endosomal or lysosomal membrane of the target cell by disrupting its membrane structure . The selected ionizable lipid ( s) may be comprised of a molar ratio of about 15-65% of the total lipid present in the formulation . In some embodiments , the ionizable lipid may be comprised in part of a dimethylammonium-propane or a derivative thereof , such as 18 : 0 DAP ( 1 , 2-distearoyl-3-dimethylammonium-propane) , 16 : 0 DAP ( 1 , 2-dipalmitoyl-3-dimethylammonium-propane) , 14 : 0 DAP ( 1 , 2-dimyristoyl-3-dimethylammonium-propane) , 18 : 1 DAP or DODAP ( 1 , 2-dioleoyl-3-dimethylammonium-propane ) . In other embodiments , the ioni zable lipid may be comprised in part of a ionizable cationic lipid such as Lipid C24 (bis ( 2-octyldodecyl) 3 , 3 ' - ( ( 4- ( 4-methylpiperazin-l-yl ) butyl ) azanediyl ) dipropionate) , ATX-001 ( 8 , 8f— [ [ 2— [ [ 2 — ( dimethylamino ) ethyl ] thio ] acetyl ] imino ] bis-octanoic acid, l , l ' -di- ( 2 Z ) -2-nonen-l-yl ester) , Lipid A9 (bis ( 2-butyloctyl) 10- (N- ( 3- ( dimethylamino ) propyl) nonanamido ) nonadecanedioate) , CIN-16645 ( 9Z , 12 Z-octadecadienoic acid, 3- [ 4 , 4-bis ( octyloxy) -1-oxobutoxy ] -2- [ [ [ [ 3— ( diethylamino) propoxy] carbonyl ] oxy] methyl ] propyl ester) , Lipid 23 ( 8- ( ( 2- ( ( 4- (dimethylamino ) butanoyl) oxy) ethyl) ( 10-oxo-10- (tridecan-7-yloxy) decyl) amino) octyl 2-hexyldecanoate ) , Lipid 29 ( 8— [ [ 8— [ ( 1 — ethylnonyl ) oxy] -8-oxooctyl ] [ 3- [ [ 2- (methylamino ) -3 , 4-dioxo-l-cyclobuten-l-yl ] amino ] propyl ] amino ] -octanoic acid, 1-octylnonyl ester) , Lipid 8 ( 4- ( dimethylamino ) -butanoic acid, 2- [ di- ( 9Z , 12 Z ) -9, 12-octadecadien-l-ylamino ] ethyl ester) , 1014 ( 15 , 19, 28-tris ( 2-hydroxytetradecyl) -15 , 19 , 24 , 28-tetraazadotetracontane-13 , 30-diol) , 246C10 ( 1 , 1 ' , 1 ' ' , 1 ' ' ' - [ 1 , 4-piperazinediylbis ( 3 , 1-propanediylnitrilo ) ] tetrakis-2-dodecanol ) , or 98N12-5 (N1 , N16-didodecyl-4 , 7 , 13-tris [ 3- (dodecylamino) -3-oxopropyl ] -4 , 7 , 10 , 13-tetraazahexadecanediamide) . In some embodiment s , the ionizable lipid may be comprised in part of an ionizable cationic lipid such as YK-009 ( 6- [ [ 4- (decyloxy) -4-oxobutyl ] ( 2-hydroxyethyl) amino ] -hexanoic acid, 2-octyldecyl ester) , CL4F8-6 ( 7- ( 4- (dipropylamino) butyl ) -7-hydroxytridecane-1 , 13-diyl bis ( 2-hexyloctanoate ) ) , IC8 ( 1 , 11, 11 1, 11 1 1- ( (piperazine-1 , 4-diylbis (propane-3 , 1-diyl ) ) bis ( azanetriyl ) ) tetrakis (tetradecan-2-ol ) ) , Lipid I II-45 ( 2-butyl-octanoic acid, 1 , 1 ' - [ [ ( 3-hydroxypropyl ) imino ] di-9, 1-nonanediyl ] ester) , ATX-100 ( 4 , 4 ' - [ [ [ [ 3- (dimethylamino ) propyl ] thio ] carbonyl ] imino ] bis-butanoic acid, 1 , 1 ' -bis ( 1-heptyloctyl ) ester) , DOG-IM4 (N- [ ( 25 Z ) -14- [ ( 9Z ) -9-octadecen-l- yloxy] -3 , 6 , 9 , 12 , 16-pentaoxatetratriacont-25-en-l-yl ] -lH-imidazole-5-carboxamide) , OF-Deg-Lin ( 9Z , 12 Z-octadecadienoic acid, 1 , 1 ' , 1 ' ' , 1 ' ' ' - [ ( 3 , 6-dioxo-2 , 5-piperazinediyl ) bis ( 4 , l-butanediylnitrilodi-2 , 1-ethanediyl ) ] ester) , or 0F-C4-Deg-Lin ( 9 , 12-octadecadienoic acid, ( 9Z , 12 Z ) -1 , 1 ' , 1 ' ' , 1 ' ' ' - [ ( 3 , 6-dioxo-2 , 5-piperazinediyl) bis ( 4 , 1-butanediylnitrilodi-4 , 1-butanediyl ) ] ester) . In yet other embodiments , the ionizable lipid may be comprised in part of an ioni zable cationic lipid such as ATX-0114 (heptadecan-9-yl ( Z ) —N— ( ( ( 4— ( dimethylamino ) butyl ) thio) carbonyl ) -N- ( 2- ( non-2-en-l-yloxy) -2-oxoethyl ) glycinate ) , L202 ( l-methyl-4-piperidinecarboxylic acid, 2— [ 9 — oxo-9- [ ( 3-pentyloctyl) oxy] nonyl ] dodecyl ester) , C13-112-tetra-tail ( 14 , 23-bis ( 2-hydroxytridecyl ) -17 , 20-dioxa-14 , 23-diazahexatriacontane-12 , 25 -diol) , OC2-K3-E10 ( 3 , 31- ( ( 2-hydroxyethyl ) azanediyl) bis (N- ( 3- (bis ( 2-hydroxydecyl) amino) propyl ) propanamide ) , RCB-4-8 (bis ( ( Z ) -12- ( ( ( non-2-yn-l-yloxy) carbonyl ) oxy) octadec-9-en-l-yl ) 3 , 3 ' - ( ( 2- ( diethylamino) ethyl) azanediyl) dipropionate ) , RM 133-3 ( ( 9Z , 91Z ) - ( ( 3 , 31- ( ( 3- ( diethylamino) propyl ) azanediyl ) bis (propanoyl ) ) bis ( oxy) ) bis ( octadec-9-ene-18 , 7-diyl) ( 5 Z , 5 ' Z , 8 Z , 8 ' Z , 11Z , 11 ' Z , 14 Z , 14 ' Z , 17 Z , 17 ' Z ) -bis ( icosa-5 , 8 , 11 , 14 , 17-pentaenoate ) ) . In yet other embodiments , the ionizable lipid may be comprised in part from an ionizable cationic lipid such as RM 137-5 ( ( 9Z , 9 ' Z ) - ( ( 3 , 3 ' - ( ( 2- ( 4-methylpiperazin-l-yl) ethyl ) azanediyl ) bis (propanoyl ) ) bis ( oxy) ) bis (octadec- 9-ene- 18 , 7-diyl) ( 4 Z , 4 ' Z , 7 Z , 7 ' Z , 10 Z , 10 ' Z , 13 Z , 13 ' Z , 16Z , 16 ' Z , 19Z , 19 ' Z ) -bis ( docosa-4 , 7 , 10 , 13 , 16 , 19-hexaenoate ) ) , PP Z-A10 (N, N ' - (piperazine-1 , 4-diylbis (propane-3 , 1-diyl) ) bis ( 3- (didecylamino) propanamide ) ) , Lipid 10 ( 4-methyl-l-piperazinepropanoic acid, 2- [di- ( 9Z , 12 Z ) -9 , 12-octadecadien-1-ylamino ] ethyl ester) , Lipid AX4 , Lipid 5 ( 8- [ ( 2-hydroxyethyl) [ 8- ( nonyloxy) -8-oxooctyl ] amino ] -octanoic acid, 1-octylnonyl ester) , TCL053 ( 2- ( ( ( 4- (dimethylamino ) butanoyl) oxy) methyl ) -2- ( ( ( ( Z ) -tetradec-9-enoyl ) oxy) methyl ) propane-1 , 3-diyl ( 9Z , 9 ' Z ) -bis (tetradec-9-enoate) ) , YSK05 ( l-methyl-4 , 4 -bis [ ( 9Z , 12 Z ) -9, 12-octadecadien-l-yloxy ] -piperidine ) , L-319 ( 9- [ 4- ( dimethylamino) -1-oxobutoxy] -heptadecanedioic acid, 1 , 17-di- (2 Z) -2-nonen-l-yl ester) , 4A3-SC8 , or C14-4 ( 1 , 1 [ [ 2- [ 2- [ 4- [ 2- [ [ 2- [ 2- [bis ( 2-hydroxytetradecyl) amino ] ethoxy] ethyl ] ( 2- hydroxytetradecyl) mino] ethyl] -1-piperazinyl ] ethoxy] ethyl] imino] bis-2-tetradecanol) . In other embodiments, the ionizable lipid may be comprised in part of DODMA ( 1 , 2-dioleyloxy-3-dimethylaminopropane) or DOBAQ (N- ( 4-carboxybenzyl) -N, N-dimethyl-2 , 3-bis (oleoyloxy) propan-l-aminium) . In some embodiments, the ionizable lipid may be comprised in part from an ionizable cationic amino lipid such as DLin-DMA (N, N-dimethyl-2 , 3-bis [ ( 9Z, 12Z) -9, 12 -octade cadi en-l-yloxy] -1-propanamine) , TT3 (Nl, N3 , N5-tris [3- (didodecylamino) propyl] -1, 3, 5-benzenetricarboxamide) , FTT5 (Hexa (octan-3-yl) 9, 9 ' , 9", 9"', 9"", 9""'- ( ( ( (benzene-1 , 3 , 5-tricarbonyl) ris (azanediyl) ) tris (propane-3, 1-diyl) ) tris (azanetriyl) ) hexanonanoate) , OF-02 (3, 6-bis [ 4- [bis [ ( 9Z, 12Z) -2-hydroxy-9, 12-octadecadien-l-yl ] amino] butyl] -2, 5-piperazinedione) , AA3-DLin (piperazine-1 , 4-diylbis (ethane-2, 1-diyl) ( 9Z, 91Z, 12Z, 121Z) -bis (octadeca-9, 12-dienoate) ) , DLin-MC3-DMA ( 4- (dimethylamino) -butanoic acid, ( 10Z, 13Z) -1- ( 9Z, 12Z) -9, 12-octadecadien-l-yl-10 , 13-nonadecadien-l-yl ester) , DLin-KC2-DMA (N, N-dimethyl-2 , 2-di- ( 9Z, 12Z ) -9, 12-octadecadien-l-yl-1, 3-dioxolane-4-ethanamine) , SM-102 (8— [ (2— hydroxyethyl) [ 6-oxo-6- (undecyloxy) hexyl] amino] -octanoic acid, 1-octylnonyl ester) , ALC-0315 ( 6- ( ( 2-hexyldecanoyl) oxy) -N- ( 6- ( (2-hexyldecanoyl) oxy) hexyl) -N- ( 4-hydroxybutyl) hexan-l-aminium) , ALC-0315 analogous-1 ( ( (2-hydroxyethyl) azanediyl) bis (octane-8 , 1-diyl) bis (2-hexyldecanoate) ) , Lipid 222 ( ( ( 4-hydroxybutyl) azanediyl) bis (octane-8, 1-diyl) bis ( 2-hexyldecanoate) ) , CL4-H6 ( 9-octadecenoic acid, 1, 1 '- [7- [ 4- ( dipropylamino) butyl] -7-hydroxy-l, 13-tridecanediyl ] ester) , AL-A12 (N-dodecyl-N- (3-methoxypropyl) dodecan-l-amine) , Lipid 14 ( ( (2- ( ( 4- (dimethylamino) butanoyl) oxy) ethyl) azanediyl) bis (octane-8, 1-diyl) bis (2-hexyldecanoate) ) , or Lipid 16 ( ( (2- ( (4- (dimethylamino) butanoyl) oxy) ethyl) azanediyl) bis (octane-8, 1-diyl) bis (2-hexyldecanoate) ) . In yet other embodiments, the ionizable lipid may be comprised in part of an ionizable cationic alkyne lipid such as Lipid A6 (di (dec-3-yn-l-yl) 9- ( ( 4- (dimethylamino) butanoyl) oxy) heptadecanedioate) or an ionizable cationic trialkyl lipid such as Lipid CL1 ( 5- (dimethylamino) -pentanoic acid, ( 6Z ) -1, 2-di- ( 4Z ) -4-decen-l-yl-6-dodecen-l-yl ester) .

[0058] In other embodiments, the ionizable lipid may be comprised in part of an ionizable disulfide-bond containing cationic lipid such as BAMEA-016B (bis (2- (dodecyldisulfaneyl) ethyl) 3, 3 ' - ( (3-methyl-9-oxo-10-oxa-13, 14-dithia-3, 6-diazahexacosyl) azanediyl) dipropionate) , Coatsome SS-E (Bis { 2- [N-methyl-N- (a-D-tocopherolhemisuccinatepropyl) amino] ethyl } disulf ide) , Coatsome SS-EC (Bis { 2- [ 4- (a-D-to copher olhemi succinate ethyl) piperidyl] ethyl } disulfide) , Coatsome SS-OB (is Coatsome SS-OC (Bis { 2- [ 4- (cis-9-octadecenoateethyl) -1-piperidinyl] ethyl } disulf ide) , Coatsome SS-OB (Bis [2- ( 4- { 2- [ 4- (cis-9-octadecenoyloxy) phenylacetoxy] ethyl }piperidinyl) ethyl] disulfide) , or analogs thereof . In some embodiments, the ionizable lipid may be comprised in part of a lipomer such as CKK-E12 (3, 6-bis [ 4- [bis (2-hydroxydodecyl) amino] butyl] -2, 5-piperazinedione) , or of a lipidoid such as C12-113 ( 1, 1 ' , 1 1 ' ' ' - [ (methylimino) bis (2, 1-ethanediylnitrilo) ] tetrakis-2-do decanol) , C12-200 (l, l '- [ [2- [4- [2- [ [2- [bis (2-hydroxydodecyl) amino] ethyl] (2-hydroxydodecyl) amino] ethyl] -1-piperazinyl] ethyl] imino] bis-2-dodecanol) , NT1-014B, 306-O12B-3 (bis (2- (octyldisulf aneyl) ethyl) 3, 31- ( (4-methyl-ll-oxo-12-oxa-15, 16-dithia- 4, 8-diazatetracosyl) azanediyl) dipropionate) , or 503-013. In yet other embodiments, the ionizable lipid may be comprised in part of a disulfide-cleavable proton-activated lipid-like material such as SS-Balm-M (tetradecanoic acid, 1, 1 '- [dithiobis [2 , 1-ethanediyl (methylimino) -3, 1-propanediyl] ] ester) or SS-BalmO-Bhe (4- [ [ ( 9Z) -l-oxo-9-octadecen-l-yl ] oxy ] -benzeneacetic acid, 1, 1 '- [dithiobis (2, 1-ethanediyl-l, 4-piperidinediyl-2 , 1-ethanediyl) ] ester) .

[0059] The formulations disclosed herein may contain a structural element comprised of at least one structural lipid, and / or at least one cationic lipid, and / or at least one polymer . The selected structural element (s) may be comprised of a molar ratio of about 0-35% of the total lipid present in the formulation . In some embodiments, the structural element may be comprised in part of a saturated phosphatidic acid such as DMBA ( 1 , 2-dimyristoyl-sn-glycero-3-phosphate) , DBBA ( 1, 2-dipalmitoyl-sn-glycero-3-phosphate) , DSBA ( 1, 2-distearoyl-sn-glycero-3-phosphate) , DLBA ( 1, 2-dilauroyl-sn-glycero-3-phosphate) , or analogs thereof . In other embodiments, the structural element may be comprised in part of an unsaturated phosphatidic acid, such as DOPA ( 1 , 2-dioleoyl-sn-glycero- 3-phosphate) , DAPA ( 1 , 2-diarachidonoyl-sn-glycero-3-phosphate) , or analogs thereof . In yet other embodiments, the structural element may be comprised in part of a phosphatidic acid with both saturated and unsaturated tails, such as POPA ( l-palmitoyl-2-oleoyl-sn-glycero-3-phosphate) , PAPA ( l-palmitoyl-2-arachidonoyl-sn-glycero-3-phosphate) , SOPAPA ( l-stearoyl-2-oleoyl-sn-glycero-3-phosphate) , SAPA (1-stearoyl- 2-arachidonoyl-sn-glycero-3-phosphate) , or analogs thereof .

[0060] In some embodiments, the structural element may be comprised in part of saturated phosphatidylcholine, such as DHPC (1, 2-dihexanoyl-sn-glycer o-3 -phospho choline

[0061]

[0062] 1, 2-diheptanoyl-sn-glycero-3-phosphocholine) , DLPC (1, 2-dilauroyl-sn-glycero-3-phosphocholine) , DMPC ( 1, 2-dimyristoyl-sn-glycero-3-phosphocholine) , DPPC ( 1, 2-dipalmitoyl-sn-glycer o-3 -phospho choline) DSPC ( 1, 2-distearoyl-sn-glycero-3-phospho choline)

[0063]

[0064] analogs thereof . In yet other embodiments, the structural element may be comprised

[0065]

[0066] part of an unsaturated phosphatidylcholine such as DOPC ( 1, 2-dioleoyl-sn-glycero-3-phosphocholine) , DLPC (1, 2-dilinoleoyl-sn-glycero-3-phosphocholine) DEPC (1, 2-dierucoyl-sn-glycero-3-phosphocholine) 1, 2-DAPC ( 1, 2- Diarachidoyl-sn-glycero-3-phosphocholine) DDPC ( 1 , 2-Didecanoyl-sn-glycero-3-phosphocholine) , 1, 2-PLPC ( l-Palmitoyl-2-Lauroyl-sn-glycero- 3 -Phospho choline)

[0067]

[0068] analogs thereof . Alternatively

[0069]

[0070] some embodiments, the structural element may be comprised of an unsaturated phosphatidylcholine with tails containing different numbers of carbon atoms such as MPPC ( 1-my ri stoy 1-2 -palmitoy 1-sn-gly cer o-3-phospho choline) MSPC ( l-myristoyl-2-stearoyl-sn-glycero-3-phospho choline) PMPC ( 1-palmit oy 1-2 -my ri stoy 1- sn-glycer o-3-phospho choline) PSPC ( 1 -palmitoy 1-2 -st e aroyl- sn-glycer o-3-phospho choline) SMPC ( 1-st ear oy 1-2 -my ri stoy 1- sn-glycer o-3-phospho choline) SPPC ( 1-st ear oy 1-2 -palmitoyl- sn-glycer o-3-phospho choline)

[0071]

[0072] analogs thereof . In yet other embodiments, the structural element may be comprised in part of a phosphatidylcholine with both saturated and unsaturated tails, such as POPC ( l-palmitoyl-2-oleoyl-glycero-3-phosphocholine) , PAPC ( l-palmitoyl-2-arachidonoyl-sn- glycero-3-phosphocholine) , SOPC ( l-stearoyl-2-oleoyl-sn-glycero-3-phosphocholine) , OMPC ( l-oleoyl-2-myristoyl-sn-glycero-3-phosphocholine) , OPPC ( l-oleoyl-2-palmitoyl-sn-glycero-3-phosphocholine) , OSPC ( l-oleoyl-2-stearoyl-sn-glycero-3-phosphocholine) , or analogs thereof . In yet other embodiments, the structural element may be comprised in part of a lysophosphatidylcholine such as S-LysoPC (l-Stearoyl-2-lyso-sn-glycero-3-phosphocholine) , P-LysoPC (l-Palmitoyl-2-lyso-sn-glycero-3-phosphocholine) , 0-LysoPC (1-01eoyl-2-lyso-sn-glycero-3-phosphocholine) , M-LysoPC ( l-Myristoyl-2-lyso-sn-glycero-3-phosphocholine) , or analogs thereof .

[0073] In some embodiments, the structural element may be comprised in part of a saturated phosphatidylethanolamine such as DLPE ( 1 , 2-dilauroyl-sn-glycero-3-phosphoethanolamine) , DPPE ( 1, 2-dipalmitoyl-sn-glycero-3-phosphoethanolamine) , DPPE-Mal (N- (3-Maleimide-l-oxopropyl) -L-a-phosphatidylethanolamine, Dipalmitoyl) , DPPE-NHS (N- (Succinimidyloxy-glutaryl) -L-a-phosphatidylethanolamine, Dipalmitoyl) , DPPE-Glu (N-Glutaryl-L-a-phosphatidylethanolamine, Dipalmitoyl) , DSPE (1, 2— distearoyl-sn-glycero-3-phosphoethanolamine) , DSPE-Mal (N- ( 3-Maleimide-l-oxopropyl) -L-a-phosphatidylethanolamine, Distearoyl) , DSPE-NHS (N- ( Succinimidyloxy-glutaryl) -L-a-phosphatidylethanolamine, Distearoyl) , DSPE-Glu (N-Glutaryl-L-a-phosphatidylethanolamine, Distearoyl) , DMPE ( 1 , 2-dimyristoyl-sn-glycero-3-phosphoethanolamine) , DMPE-Mal (N- (3-Maleimide-l-oxopropyl) -L-a-phosphatidylethanolamine, Dimyristoyl) , DMPE-NHS (N- (Succinimidyloxy-glutaryl) -L-a-phosphatidylethanolamine, Dimyristoyl) , DMPE-Glu (N-Glutaryl-L-a-phosphatidylethanolamine, Dimyristoyl) or an analog thereof . In other embodiments, the structural element may be comprised in part of an unsaturated phosphatidylethanolamine, such as DOPE ( 1 , 2-dioleoyl-sn-glycero-3-phosphoethanolamine) , DOPE-Mal (N- (3-Maleimide-l-oxopropyl) -L-a-phosphatidylethanolamine, Dioleoyl) , DOPE-NHS (N- ( Succinimidyloxy-glutaryl) -L-a-phosphatidylethanolamine, Dioleoyl) , DOPE-Glu (N-Glutaryl-L-a-phosphatidylethanolamine, Dioleoyl) , DAPE (1, 2— diarachidonoyl-sn-glycero-3-phosphoethanolamine) , or analogs thereof . In yet other embodiments, the structural element may be comprised in part of a phosphatidylethanolamine with both saturated and unsaturated tails, such as POPE ( l-palmitoyl-2-oleoyl-sn-glycero-3-phosphoethanolamine) , SOPE ( l-stearoyl-2-oleoyl-sn-glycero-3-phosphoethanolamine) , PAPE ( l-palmitoyl-2-arachidonoyl-sn-glycero-3-phosphoethanolamine) , SAPE ( l-stearoyl-2-arachidonoyl-sn-glycero-3-phosphoethanolamine) , or an analog thereof .

[0074] In yet other embodiments, the structural element may be comprised in part of a saturated phosphatidylglycerol such as DLPG ( 1 , 2-dilauroyl-sn-glycero-3-phospho- ( 1 ' -rac-glycerol) ) , DMPG ( 1 , 2-dimyristoyl-sn-glycero-3-phospho- (1 ' -rac-glycerol) ) , DPPG ( 1, 2-dipalmitoyl-sn-glycero-3-phospho- (1 ' -rac-glycerol) ) , DSPG ( 1, 2-distearoyl-sn-glycero-3-phospho- (1 ' -rac-glycerol) ) , DOPG (1, 2-dioleoyl-sn-glycero-3-phospho- ( 1 ' -rac-glycerol) ) , or analogs thereof . In other embodiments, the structural element may be comprised in part of an unsaturated phosphatidylglycerol or a phosphatidylglycerol with both saturated and unsaturated tails, such as POPG ( l-palmitoyl-2-oleoyl-sn-glycero-3-phospho- (1 ' -rac-glycerol) ) , PAPG ( l-palmitoyl-2-arachidonoyl-sn-glycero-3-phospho- (1 ' -rac-glycerol) ) , SOPG ( l-stearoyl-2-oleoyl-sn-glycero-3-phospho- (1 ' -rac-glycerol) ) , SAPG ( l-stearoyl-2-arachidonoyl-sn-glycero-3-phospho- ( 1 ' -rac-glycerol) ) , or analogs thereof .

[0075] In some embodiments, the structural element may be comprised in part of a saturated phosphatidylserine, such as DSPS ( 1 , 2-distearoyl-sn-glycero-3-phospho-L-serine) , DMPS ( 1 , 2-dimyristoyl-sn-glycero-3-phospho-L-serine) , DPPS ( 1 , 2-dipalmitoyl-sn-glycero-3-phospho-L-serine) , DLPS ( 1, 2-dilauroyl-sn-glycero-3-phospho-L-serine) , or analogs thereof . In other embodiments, the structural element may be comprised in part of an unsaturated phosphatidylserine such as DOPS ( 1 , 2-dioleoyl-sn-glycero-3-phospho-L-serine) or an analog . In yet other embodiments, the structural element may be comprised in part of a phosphatidylserine with both saturated and unsaturated tails, such as POPS ( l-palmitoyl-2-oleoyl-sn-glycero-3-phospho-L-serine) , SOPS ( l-stearoyl-2-oleoyl-sn-glycero-3-phospho-L-serine) , PAPS ( l-palmitoyl-2-arachidonoyl-sn-glycero-3-phospho-L-serine) , SAPS ( l-stearoyl-2-arachidonoyl-sn- glycero-3-phospho-L-serine) , or analogs thereof .

[0076] In general, the formulations disclosed herein may be comprised in part of additional nonionizable cationic lipids . These lipids generally are expected to have a permanent positive charge at physiological pH . In some embodiments, the cationic lipid may be comprised in part of DORI (N- ( 2-hydroxyethyl) -N, N-dimethyl-2 , 3 -bis (oleoyloxy) propan-l-aminium bromide) , DC-6-14 (0, O' -ditetradecanoyl-N- (a-trimethylammonioacetyl) diethanolamine) , DSDMA ( 1 , 2-distearyloxy-N, N-dimethyl-3-aminopropane) , DMRIE (1, 2-dimyristyloxypropyl-3-dimethyl-hydroxyethyl ammonium bromide) , or analogs thereof . In other embodiments, the cationic lipid may be comprised in part of a quaternary ammonium compound such as 18 : 0 DDAB (Dimethyldioctadecylammonium) , DC-1-16 (N-Hexadecyl-N, N-dimethylhexadecan-l-aminium) , dimethyldimyristylammonium (N, N-dimethyl-N-tetradecyl-1-tetradecanaminium) , DODAC N, N-Dioleyl-N, N-dimethylammonium chloride) , or analogs thereof . In yet other embodiments, the cationic lipid may be comprised in part of a trimethylammoniumpropane, such as 14 : 0 TAP (1, 2— dimyristoyl-3-trimethylammoniumpropane) , 16 : 0 TAP ( 1 , 2-dipalmitoyl-3-trimethylammoniumpropane) , 18 : 0 TAP ( 1, 2-stearoyl-3-trimethylammoniumpropane) , or analogs thereof, or more preferentially, 18 : 1 TAP (DOTAP, 1, 2-dioleoyl-3-trimethylammoniumpropane) , DOTMA (1, 2 — di-0-octadecenyl-3-trimethylammonium propane) , or analogs thereof .

[0077] In some embodiments, the cationic lipid may be comprised in part of an esterified phosphatidylcholine, such as 12 : 0 EPC ( 1, 2-dilauroyl-sn-glycero-3-ethylphosphocholine) , 14 : 0 EPC ( 1 , 2-dimyristoyl-sn-glycero-3-ethylphosphocholine) , 16 : 0 EPC ( 1, 2-dipalmitoyl-sn-glycero-3-ethylphosphocholine) , 18 : 0 EPC ( 1 , 2-distearoyl-sn-glycero-3-ethylphosphocholine) , 14 : 1 EPC ( 1 , 2-dimyristoleoyl-sn-glycero-3-ethylphosphocholine) , 18 : 1 EPC ( 1 , 2-dioleoyl-sn-glycero-3-ethylphosphocholine) , 16 : 0-18 : 1 EPC ( l-palmitoyl-2-oleoyl-sn-glycero-3-ethylphosphocholine) , or analogs thereof . In other embodiments, the cationic lipid may be comprised in part of a multivalent cationic lipid, such as MVL5 (Nl- [2- ( (IS) -1- [ (3-aminopropyl) amino] -4- [di (3-amino- propyl) amino] butylcarboxamido) ethyl] -3, 4-di [oleyloxy] -benzamide) or an analog . In yet other embodiments, the cationic lipid may be comprised in part of a spermine-derive lipid, such as DOSPA ( 2 , 3-dioleyloxy-N- [ 2- ( sperminecarboxamido) ethyl ] -N, N-dimethyl-l-propanaminium) , GL67 (N4-Cholesteryl-Spermine) , DOGS (Dioctadecylamidoglycylspermine) , DPPES (Dipalmitoylphosphatidylethanolamidospermine) , RPR120535, or analogs thereof, or it may be comprised in part of a cholesterol-based cationic lipid such as DC-Cholesterol • HC1 ( 3f>- [N- (N ' , N ' -dimethylaminoethane) -carbamoyl] cholesterol hydrochloride) , CLin-DMA (3-dimethylamino-2- ( cholest-5-en-3-beta-oxybutan-4-oxy) -1- (cis, cis-9, 12-octadecadienoxy) propane) , or analogs thereof .

[0078] In general, the formulations disclosed herein may be comprised in part of polymers . The selected polymer (s) may be comprised of a molar ratio of about 0-20% of the total lipid present in the formulation . In some embodiments, the polymer may be comprised in part of a polyethylene glycol (PEG) -modified lipid with a polyethylene glycol chain of up to 350 Da in length, such as 14 : 0 PEG350 PE ( 1 , 2-dimyristoyl-sn-glycero-3-phosphoethanolamine-N- [methoxy (polyethylene glycol) -350 ] ) , 16 : 0 PEG350 PE ( 1, 2-dipalmitoyl-sn-glycero-3-phosphoethanolamine-N- [methoxy (polyethylene glycol) -350 ] ) , 18 : 0 PEG350 PE ( 1 , 2-distearoyl-sn-glycero-3-phosphoethanolamine-N- [methoxy (polyethylene glycol) -350 ] ) , 18 : 1 PEG350 PE ( 1 , 2-dioleoyl-sn-glycero-3-phosphoethanolamine-N- [methoxy (polyethylene glycol) -350 ] ) , or analogs thereof . In other embodiments, the polymer may be comprised in part of a PEG-modified lipid with a polyethylene glycol chain of up to 550 Da in length, such as 14 : 0 PEG550 PE (DMPE-PEG550 , 1, 2-dimyristoyl-sn-glycero-3-phosphoethanolamine-N- [methoxy (polyethylene glycol) -550 ] ) , 16 : 0 PEG550 PE ( 1, 2-dipalmitoyl-sn-glycero-3-phosphoethanolamine-N- [methoxy (polyethylene glycol) -550 ] ) , 18 : 0 PEG550 PE (DSPE-PEG550 , 1, 2-distearoyl-sn-glycero-3-phosphoethanolamine-N- [methoxy (polyethylene glycol) -550 ] ) , 18 : 1 PEG550 PE (DOPE-PEG550 , 1 , 2-dioleoyl-sn-glycero-3-phosphoethanolamine-N- [methoxy (polyethylene glycol) -550 ] ) , or analogs thereof . In yet other embodiments, the polymer may be comprised in part of a PEG-modified lipid with a polyethylene glycol chain of up to 750 Da in length, such as 14 : 0 PEG750 PE ( 1, 2-dimyristoyl-sn-glycero-3-phosphoethanolamine-N- [methoxy (polyethylene glycol) -750 ] ) , 16 : 0 PEG750 PE ( 1, 2-dipalmitoyl-sn-glycero-3-phosphoethanolamine-N- [methoxy (polyethylene glycol) -750 ] ) , 18 : 0 PEG750 PE ( 1 , 2-distearoyl-sn-glycero-3-phosphoethanolamine-N- [methoxy (polyethylene glycol) -750 ] ) , 18 : 1 PEG750 PE ( 1 , 2-dioleoyl-sn-glycero-3-phosphoethanolamine-N- [methoxy (polyethylene glycol) -750 ] ) , or analogs thereof . In yet other embodiments, the polymer may be comprised in part of a PEG-modified lipid with a polyethylene glycol chain of up to 1 kDa in length, such as 14 : 0 PEG1000 PE (DMPE-PEG1000 , 1, 2-dimyristoyl-sn-glycero-3-phosphoethanolamine-N- [methoxy (polyethylene glycol) -1000 ] ) , 16 : 0 PEG1000 PE (DPPE-PEG1000 , 1 , 2-dipalmitoyl-sn-glycero-3-phosphoethanolamine-N- [methoxy (polyethylene glycol) -1000 ] ) , 18 : 0 PEG1000 PE (DSPE-PEG1000 , 1, 2-distearoyl-sn-glycero-3-phosphoethanolamine-N- [methoxy (polyethylene glycol) -1000 ] ) , 18 : 1 PEG1000 PE (DGPE-PEG1000 , 1 , 2-dioleoyl-sn-glycero-3-phosphoethanolamine-N- [methoxy (polyethylene glycol) -1000 ] ) , or analogs thereof . In some embodiments, the polymer may be comprised in part of a PEG-modified lipid with a polyethylene glycol chain of up to 2 kDa in length, such as 14 : 0 PEG2000 PE (DMPE-PEG2000, 1 , 2-dimyristoyl-sn-glycero-3-phosphoethanolamine-N- [methoxy (polyethylene glycol) -2000 ] ) , 16 : 0 PEG2000 PE (DPPE-PEG2000 , 1 , 2-dipalmitoyl-sn-glycero-3-phosphoethanolamine-N- [methoxy (polyethylene glycol) -2000 ] ) , 18 : 0 PEG2000 PE (DSPE-PEG2000 , 1, 2-distearoyl-sn-glycero-3-phosphoethanolamine-N- [methoxy (polyethylene glycol) -2000 ] ) , DSPE-2arm-PEG2000 (N- [2 ' , 3 ' -Bis (methylpolyoxyethylene oxy) propane-1 ' -oxycarbonyl] -1, 2-distearoyl-sn-glycero-3-phosphoethanolamine) , DSPE-PEG2K-triGalNAc ( 1 , 2-distearoyl-sn-glycero-3-phosphoethanolamine-N- [carboxy (polyethylene glycol) -2000 ] -N-tri (N-acetylgalactosamine) ) , 18 : 1 PEG2000 PE (DGPE-PEG2000 , 1 , 2-dioleoyl-sn-glycero-3-phosphoethanolamine-N- [methoxy (polyethylene glycol) -2000 ] ) , or analogs thereof . In other embodiments, the polymer may be comprised in part of a PEG-modified lipid with a polyethylene glycol chain of up to 3 kDa in length, such as 14 : 0 PEG3000 PE ( 1 , 2-dimyristoyl-sn-glycero-3-phosphoethanolamine-N- [methoxy (polyethylene glycol) -3000 ] ) , 16 : 0 PEG3000 PE ( 1, 2-dipalmitoyl-sn-glycero-3-phosphoethanolamine-N- [methoxy (polyethylene glycol) -3000 ] ) , 18 : 0 PEG3000 PE ( 1, 2-distearoyl-sn-glycero-3-phosphoethanolamine-N- [methoxy (polyethylene glycol) - 3000 ] ) , 18 : 1 PEG3000 PE ( 1, 2-dioleoyl-sn-glycero-3-phosphoethanolamine-N- [methoxy (polyethylene glycol) -3000 ] ) , or analogs thereof . In yet other embodiments, the polymer may be comprised in part of a PEG-modified lipid with a polyethylene glycol chain of up to 5 kDa in length, such as 14 : 0 PEG5000 PE ( 1 , 2-dimyristoyl-sn-glycero-3-phosphoethanolamine-N- [methoxy (polyethylene glycol) -5000 ] ) , 16 : 0 PEG5000 PE ( 1 , 2-dipalmitoyl-sn-glycero-3-phosphoethanolamine-N- [methoxy (polyethylene glycol) - 5000 ] ) , 18 : 0 PEG5000 PE ( 1 , 2-distearoyl-sn-glycero-3-phosphoethanolamine-N- [methoxy (polyethylene glycol) -5000 ] ) , DSPE-2arm-PEG5000 (N- [2 ' , 3 ' -Bis (methylpolyoxyethylene oxy) propane-1 ' -oxycarbonyl] -1, 2-distearoyl-sn-glycero-3-phosphoethanolamine) , 18 : 1 PEG5000 PE ( 1, 2-dioleoyl-sn-glycero-3-phosphoethanolamine-N- [methoxy (polyethylene glycol) -5000 ] ) , or analogs thereof .

[0079] In some embodiments, the polymer may be comprised in part of a PEG-modified glyceride, such as DMG-PEG2000 ( 1 , 2-dimyristoyl-rac-glycero-3-methoxypolyethylene glycol-2000) , DSG-PEG2000 (a- [2 , 3-bis [ ( 1-oxooctadecyl) oxy] propyl] -w-methoxy-poly (oxy-1, 2-ethanediyl) ) , DPG- PEG2000 ( 1, 2-Dipalmitoyl-rac-glycero-3-methylpolyoxyethylene) , DOG- PEG2000 ( 1, 2-Dioleoyl-rac-glycerol, methoxypolyethylene Glycol) , DMG-PEG5000 ( 1, 2-Dimyristoyl-rac-glycero-3-methylpolyoxyethylene) , DSG- PEG5000 ( 1, 2-Distearoyl-rac-glycero-3-methylpolyoxyethylene) , DPG- PEG5000 ( 1 , 2-Dipalmitoyl-rac-glycerol, methoxypolyethylene Glycol) , DGG-PEG5000 (1, 2-Dioleoyl-rac-glycerol, methoxypolyethylene Glycol) , Sunbright GS-020MA (PEG2000, 1 , 2-Distearoyl-rac-glycero-3- ( 2 ' -maleimidoethyl) polyoxyethylene) , Sunbright GS-020TS (PEG2000 , 1, 2- Distearoyl-rac-glycero-3- ( succinimidyloxycarbonyl) polyoxyethylene) , ALC-0159 (Methoxypolyethyleneglycoloxy (2000) -N, N-ditetradecylacetamide) , or analogs thereof . In other embodiments, the polymer may be comprised in part of a PEG-modified sterol such as Chol-PEG600 (Cholesterol- (polyethylene glycol-600) ) , Chol-PEG2000 (Cholesterol- (polyethylene glycol-2000) ) , or analogs thereof, or a PEG- modified ceramide such as C8 PEG750 ceramide (N-octanoyl-sphingosine-1- { succinyl [methoxy (polyethylene glycol) 750 ] } ) , C16 PEG750 ceramide (N-palmitoyl-sphingosine-1- { succinyl [methoxy (polyethylene glycol) 750 ] } ) , C8 PEG2000 ceramide (N-octanoyl-sphingosine-1- { succinyl [methoxy (polyethylene glycol) 2000 ] } ) , C16 PEG2000 ceramide (N-palmitoyl-sphingosine-1- { succinyl [methoxy (polyethylene glycol) 2000 ] } ) , C8 PEG5000 ceramide (N-octanoyl-sphingosine-1- { succinyl [methoxy (polyethylene glycol) 5000 ] } ) , C16 PEG5000 ceramide (N-palmitoyl-sphingosine-1- { succinyl [methoxy (polyethylene glycol) 5000 ] } ) , or analogs thereof .

[0080] In other embodiments, the polymer may be a comprised in part of diacetylene lipid such as 23 : 2 Diyne PE ( 1, 2-bis ( 10 , 12-tricosadiynoyl) -sn-glycero-3-phosphoethanolamine) , 16 : 0-23 : 2 Diyne PE (PTPE, 1-palmitoyl-2- (10, 12-tricosadiynoyl) -sn-glycero-3-phosphoethanolamine) , 23 : 2 Diyne PC ( 1, 2-bis ( 10 , 12-tricosadiynoyl) -sn-glycero-3-phosphocholine) , 16 : 0-23 : 2 Diyne PC (PTPC, l-palmitoyl-2- ( 10 , 12-tricosadiynoyl) -sn-glycero-3-phosphocholine) , or analogs thereof . In yet other embodiments, the polymer may be comprised in part of Tocofersolan (TPGS 1000, D-a-tocopheryl polyethylene glycol succinate) or an analog.

[0081] In general, the formulations disclosed herein may be comprised in part of intracellular vesicle-interacting molecules, including those that may interact with endosomes, caveosomes, macropinosomes, phagosomes, or other intracellular vesicles . These intracellular vesicle-interacting molecules may be cell-penetrating molecules or peptides, membrane-disruptive peptides, endosome-disrupting peptides, endosomal escape vehicles, fusogenic peptides, an amphiphilic peptide, a miniature protein, peptides containing an endosomal escape domain or a penetration accelerating sequence, or generally a molecule, compound, peptide, or nucleic acid that disrupts intracellular vesicle membranes to allow contents out into the cytosol . In some embodiments, the intracellular vesicle-interacting peptide may be comprised in part of a cellpenetrating molecule such as CPM2, or a membrane-disruptive peptide such as aurein 1.2 (GLFDIIKKIAESF) , or analogs thereof . In other embodiments, the intracellular vesicle-interacting peptide may be comprised in part of an endosome-disrupting peptide truncated form of the N-terminus of adenovirus protein VI (LLLLAFSWGSLWSGIKNFGSTIKN) , Tat ( YGRKKRRQRRR) , dTat-HA2 (D-Tat fused to HA2) , transportan 10 (AGYLLGKINLKALAALAKKIL) , octaarginine (R8) , nonaarginine (R9) , octalysine (L8 ) , penetratin (RQIKIWFQNRRMKWKK) , or analogs thereof . In yet other embodiments, the intracellular vesicle-interacting peptide may be comprised in part of a histidine-swit ching cell-penetrating peptide such as hsTAT (YGHKKHHQHHH) , hsTPIO (FSGSGSGSGAGYLLGHINLHALAALAHHILGC) , hsPen (FSGSGSGSGHQIHIWFQNHHMHWHHGC) , hsTPT ( CLIHHALAALAHLNIHLLYGASNLTWG) , hsLMWP (GSVSHHHHHHGGHHHH) , hsR12 (HHHHHHHHHHHH) , hsMPG (GALFLGFLGAAGSTMGAWSQPHHHHHV) , or analogs thereof .

[0082] In some embodiments, the intracellular vesicle-interacting peptide may be comprised in part of a fusogenic peptide such as CM18 , CM18-Tat (KWKLFKKIGAVLKVLTTG-YGRKKRRQRRR) , diINF-7, H5WYG, or analogs thereof . In other embodiments, the intracellular vesicle-interacting peptide may be comprised in part of a pH-sensitive fusogenic peptide such as HA2 (GLFGAIAGFIENGWEGMIDGWYG) , GALA (WEAALAEALAEALAEHLAEALAEALEALAA) , or analogs thereof . In yet other embodiments, the intracellular vesicleinteracting peptide may be comprised in part of an amphiphilic peptide such as 6His-CM18-PTD4 , pHlip, pHD118, or analogs thereof . In yet other embodiments, the intracellular vesicle-interacting peptide may be comprised in part of a cyclic cell-penetrating peptide such as cF4>R4 (cyclic phenylalanine-L-2-naphthylalanine-tetraarginine) , CPP9, CPP12 , palmitoyl-cyclic-dodeca-D-arginine (palmitoyl-GSSGCRRRRRRRRRRRRC-NH2 ) , or analogs thereof . In some embodiments, the intracellular vesicleinteracting peptide may be comprised in part of a miniature protein such as 5.3, ZF5.3, or analogs thereof . In some alternate embodiments, the intracellular vesicle-interacting peptide may be comprised in part of a peptide containing an endosomal escape domain or a penetration accelerating sequence such as GKPILFF or an analog .

[0083] In general, the formulation disclosed in this invention may be comprised in part of sterols, cholesterols, or other cholesterol-like molecules that modulate the lipid-packing and fluidity of the LNP formulation . The selected sterol ( s) , cholesterol ( s ) , or cholesterol-like molecule ( s) may be comprised of a molar ratio of about 0-55% of the total lipid present in the formulation . In some embodiments, the cholesterol or cholesterol analog may be comprised in part of cholesterol ( (3p) -cholest-5-en-3-ol) , desmosterol ( 3f>-hydroxy-5 , 24-cholestadiene) , lanosterol (lanosta-8, 24-dien-3p-ol) , lanosterol-95 ( 8, 24-lanostadien-3b-ol) , 14-demethyl-lanosterol ( 4 , 4-dimethyl cholest-8 ( 9) , 24-dien-3f>-ol) , zymosterol (5a-cholesta-8 , 24-dien-3f>-ol) , lathosterol ( 5a-cholest-7-en-3f>-ol) , 24-methylene cholesterol (5, 24 (28 ) -cholestadien-24-methylen-3f>-ol) , or analogs thereof . In other embodiments, the cholesterol or cholesterol analog may be comprised in part of a dehydrosterol such as 7-dehydrocholesterol (A5, 7-cholesterol) , 8 ( 9) -dehydrocholesterol ( cholesta-5, 8 ( 9) -dien-3f>-ol) , 8 ( 14 ) -dehydrocholesterol (cholesta-5, 8 (14) -dien-3f>-ol) , FF-MAS (14-demethyl-14-dehydrolanosterol) , or analogs thereof . In yet other embodiments, the cholesterol or cholesterol analog may be comprised in part of a saturated sterol such as dihydrolanosterol (24 , 25-dihydrolanosterol) , zymostenol (5 a -Cholest-8-en-3-beta-ol) , or analogs thereof .

[0084] In some embodiments, the cholesterol or cholesterol analog may be comprised in part of a cholesterol ester such as cholesteryl nervonate (cholest-5-en-3-ol, 3p- [ ( 15Z) -15-tetracosenoate] ) , cholesteryl oleate (cholest-5-en-3p-ol, 3- [ ( 9Z ) -9-octadecenoate ] ) , cholesteryl palmitate ( (3p) -cholest-5-en-3-ol, 3-hexadecanoate) , 17 : 0 cholesteryl ester (cholest-5-en-3f>-yl heptadecanoate) , cholesteryl stearate ( (3p) -cholest-5-en-3-ol, 3-octadecanoate) , cholesteryl lignocerate (cholest-5-en- (3p) -ol, 3-tetracosanoate) , cholesteryl homo-y-linolenate (cholest-5-en-3p-ol, 3- ( 8Z, llZ, 14Z) -8, ll, 14-eicosatrienoate) , or analogs thereof . In other embodiments, the cholesterol or cholesterol analog may be comprised in part of an oxysterol such as 4 p-hydroxy cholesterol (cholest-5-ene-3p, 4p-diol) , 25-hydroxy cholesterol (cholest-5-ene-3p, 25-diol) , or analogs thereof .

[0085] In other embodiments, the cholesterol or cholesterol analog may be comprised in part of a phytosterol (derived from plants) , such as stigmasterol ( stigmasta-5 , 22E-dien-3p-ol) , 24-methyl cholesterol ( ( 3p, 24 -ergost-5-en-3-ol) , p-S it o sterol ( stigmast-5-en-3p-ol) , brassicasterol (ergosta-5, 22E-dien-3p-ol) , fucosterol ( (3p, 24E) -stigmasta-5, 24 (28) -dien-3-ol) , a-spinasterol ( (3p, 5a, 22E) -stigmasta-7 , 22-dien-3-ol) , campesterol (campest-5-en-3f>-ol) , or analogs thereof . In yet other embodiments, the cholesterol or cholesterol analog may be comprised in part of a saturated phytosterol, such as sitostanol ( ( 5a) -stigmastan-3p-ol) , campestanol ( (3p, 5a, 24R) -ergostan-3-ol) , or analogs thereof .

[0086] The nanoparticle formulations may contain a targeting element . The targeting element may be an antibody, antibody fragment, nanobody, antigen-binding fragment, protein, peptide, receptor ligand, receptor binding domain, lipid, polysaccharide, nucleic acid, aptamer, or other targeting element that aids the nanoparticle in accumulating in a particular tissue type, or binding to and entering a specific type of cell . In some embodiments, the targeting element may be comprised in part of an antibody, such as anti-c-Kit (anti-CD117) , antiglucocorticoid receptor, anti-transforming growth factor beta receptor 2 (TGFBR2) , anti-transforming growth factor beta receptor 3 (TGFBR3) , anti-aquaporin 7 , anti-aquaporin 7B, anti-activin A receptor type 1C, anti-frizzled class 4 receptor, anti-perilipin 1, anti-perilipin 4, anti-SLC7A10, anti-platelet-derived growth factor receptor alpha (PDGFR-alpha) , anti-platelet-derived growth factor receptor beta (PDGFR-beta) , or analogs thereof . In other embodiments, the targeting element may be comprised in part of a nanobody or antigen-binding fragment (Fab) , which are much smaller than antibodies . In yet other embodiments, the targeting element may be comprised in part of a protein such as angiotensin II, VEGF, wheat germ agglutinin (WGA) , Ulex europaeus agglutinin I (UEA-1) , miniAp-4, PEP3, or analogs thereof . In other embodiments, the targeting element may be comprised in part of a peptide sequence such as CKGGRAKDC, GGGGYDRVTIHPF, D (KLAKLAK)2, MRYMILGLLALAAVCSA, or analogs thereof .

[0087] In some embodiments, the targeting element may be comprised in part of a receptor ligand capable of binding to a specific receptor on the cell surface, such as LJ-4378 (agonist of A2A adenosine receptor / antagonist of A3 adenosine receptor) , resistin (ligand of decorin lacking the glycanation site) , norrin ( specific ligand of frizzled class 4 receptor) , or analogs thereof . Alternatively, the targeting element may be comprised in part of a lipid such as PAHSA (palmitic-acid-9-hydroxy-stearic-acid) or an analog, or of a polysaccharide such as glycogen, glucan, dextran, chitosan, alginate, hyaluronic acid or analogs thereof . In yet other embodiments, the targeting element may be a nucleic acid or an aptamer, such as an aptamer against VEGFR.

[0088] The lipid nanoparticle formulations described herein can be intended to deliver a deoxyribonucleic acid (DNA) cargo to cells for the treatment of disease . The DNA may be single-stranded or double-stranded, linear or circular, and may be 0. 5 Kb to 50 Kb in length, or ideally 1.5 Kb to 7 .0 Kb in length . The genetic cargo may encode at least a promoter and / or enhancer . The promoter may be a general promoter, such as the cytomegalovirus (CMV) immediate early promoter, the eukaryotic translation elongation factor 1 al (EF1A) promoter, the eukaryotic translation elongation factor 1 al short form (EFS) promoter, the CAG promoter, the CBh promoter, the CBA promoter, simian virus 40 (SV40 ) enhancer / early promoter, or the phosphoglycerate kinase 1 (BGK) promoter . Alternatively, the promoter may be a tissue-specific promoter; for adipose, this may be the adipocyte B2 (aP2 ) promoter, the adiponectin promoter, the peroxisome prolif erator-activated receptor gamma (PEAR gamma) promoter, the fatty acid binding protein 4 (FABP4) promoter, the laeverin (LVRN) promoter, the perilipin 1 (PLIN1) promoter, or the perilipin 4 (PLIN4 ) promoter .

[0089] A portion of the genetic cargo can encode a therapeutic transgene, which is intended to effectively treat disease in embodiments . In some embodiments, the therapeutic transgene may be intended to treat diabetes, for example, the therapeutic transgene may encode at least insulin, gastric inhibitory peptide (GIP) receptor agonist, glucagon-like peptide 1 (GLP1) receptor agonist, a GIP / GLP1 receptor co-agonist, or analogs thereof . In other embodiments, the therapeutic transgene may encode at least insulin-like growth factor ( IGF) 1, IGF2 , or analogs thereof to treat growth restriction . In yet other embodiments, the therapeutic transgene may encode at least beta-glucocerebrosidase for the treatment of Gaucher' s Disease or alpha-glucosidase for the treatment of Pompe Disease .

[0090] In some other embodiments, the therapeutic transgene may encode at least fibroblast growth factor 18 (FGF18 ) , vascular endothelial growth factor (VEGFA) , factor VIII, factor IX, or liver-expressed antimicrobial peptide 2 (LEAP2 , MTPFWRGVSLRPIGASCRDDSECITRLCRKRRCSLSVAQE) . In other embodiments, the therapeutic transgene may encode at least an siRNA, such as one against X-linked inhibitor of apoptosis-associated factor 1 (XAF1) or an shRNA, antibody, antibody fragment, or a nanobody such as an anti-tumor necrosis factor alpha (TNF-alpha) nanobody, capable of reducing expression of a gene or function of a protein known to cause disease .

[0091] The genetic cargo may optionally include a post-transcriptional regulatory element to aid in transduction efficiency, such as the Hepatitis B post-transcriptional regulatory element (HPRE) , the Woodchuck Hepatitis Virus post-transcriptional regulatory element (WPRE) , or the optimized Woodchuck Hepatitis Virus post-transcriptional regulatory element (oPRE) . The genetic cargo may also optionally include a polyadenylation sequence, such as the bovine growth hormone polyadenylation sequence (bGH pA) or the SV40 polyadenylation sequence (SV40 pA) , to prevent unwanted readthrough of genes . Furthermore, the genetic cargo may optionally include a downtitration element, that is, and element to adjust the therapeutic dose downwards after administration, such as a suicide gene (e . g. inducible caspase 8 [iCasp8 ] or inducible caspase 9 [iCasp9] ) .

[0092] The components described herein can optionally work in concert to enable any functionality of the system or more therapeutic elements, but may also optionally have additional functionality such as increased persistence in the circulation, improved biocompatibility, improved bioresorption, optimized biodegradation, or may impact other cellular elements or pathways, or interact with other cellular sequences, proteins, organelles, or function .

[0093] It should be understood that within the scope of this invention the formulation, materials, genetic constructs, sequences, biological and chemical compositions, or methods of use may be varied by one skilled in the art, to the extent that the structures described herewithin perform the desired function and remain within the scope of the present invention . Various parts, components or characteristics may be used in combination, with or without modification by someone skilled in the art to achieve the desired functionality of the af oredescribed formulation . Moreover, all individual features and methods of use described herein, and each and every combination of two or more of such features and methods of use, are included within the scope of the present invention provided that these features and methods of use in such a combination are not mutually inconsistent . It is understood that certain portions or combinations of such portions can be varied by someone trained in the art while still achieving the main goal of the invention .

[0094] Finally, it is understood that the specific ranges provided in the current invention are not restrictive and are for example purposes only, values outside of the specified ranges may be used to achieve the goal of the invention without modification to the proposed mechanistic principals .

[0095] EXAMPLES

[0096] Example 1

[0097] A lipid nanoparticle formulation was prepared with an N to P ratio of 7 and containing a therapeutic transgene (GLP1 receptor agonist) for the treatment of diabetes . * Molar Lipid Formal Name

[0098] ratio, % 8- [ (2-hydroxyethyl) [ 6-oxo-6-1

[0099] SM-102 (undecyloxy) hexyl ] amino ] -octanoic acid, 1- 50

[0100] octylnonyl ester

[0101] DSPC 1 , 2 -di st ear oyl-sn-glycero-3 -phospho choline 10

[0102] Cholesterol (3p) -cholest-5-en-3-ol 38. 5

[0103] 1, 2-dimyristoyl-rac-glycero-3- DMG-PEG2000 1 . 5 methoxypolyethylene glycol-2000

[0104]

[0105] The formulation N to P ratio resulted in optimal transfection efficiency of primary human adipocytes relative to the other formulations tested. Similarly, the formulation created an optimal encapsulation efficiency and can ensure sufficient genetic cargo load to produce a downstream reporter gene or therapeutic effect .

[0106] dsDNA

[0107] Particle

[0108] LNP payload Encapsulation STD N / P size PDX potential

[0109] XD size (mV)

[0110] (ma) (mV)

[0111] (nt)

[0112] 3 51 85. 1 0 .17 -10 .8 1. 9 6 94 85. 8 0 .08 -5.0 1. 5 LNP

[0113] Test 7 99 91.7 0.08 1.8 RMD

[0114] payload 8 99 88. 6 0 .06 -2 .5 i . 3 P101- of 3151 10 99 89.3 0 .19 -7 .2 0.3 102

[0115] 20 100 85. 8 0 .18 -1.2 2. 0 30 100 81. 6 0 .18 — —

[0116]

[0117] The formulation was found to be robust and stable over a broad range of genetic cassette sizes, with all critical to function physico-chemical remaining effectively unchanged. These factors enable the formulation to be used as a flexible technological platform for DNA delivery to adipocytes with a high transfection efficiency, optimal cytocompatibility and high encapsulation efficiency.

[0118] The formulation was composed of genetic cargo encapsulated in LNPs that were suspended in phosphate-buf fered saline (PBS) . The formulation was administered via a subcutaneous injection at a dose of 625 ug / kg body weight at t=0 or at a dose of 312 .5 ug / kg at t=0 and t=l week . The formulation was injected into high fat diet and streptozotocin induced diabetic mice and the animals were monitored for weight loss, insulin sensitivity using the insulin tolerance test ( ITT) , and glucose sensitivity using the glucose tolerance test (GTT) . Following 4 weeks, weight loss was observed, which is comparable to daily EX4 injections and is statistically superior to the high fat diet (HFD) negative control and low-fat diet (LFD) positive control . Similarly, improved insulin tolerance and glucose response were observed relative to the negative control (HFD) .

[0119] Example 2

[0120] A lipid nanoparticle formulation is prepared using a biodegradable lipid with an N to P ratio of 7 and contains a therapeutic transgene (IGF2) for the treatment of Silver-Russell Syndrome .

[0121] Lipid Formal Name Molar ratio, % Coatsome SS-OP n / a 50

[0122] 1, 2-distearoyl-sn-glycero-3- DSPC 10

[0123] phospho choline

[0124] Cholesterol (3p) -cholest-5-en-3-ol 38 .5

[0125] 1, 2-dimyristoyl-rac-glycero-3- DMG-PEG2000 1. 5

[0126] methoxypolyethylene glycol-2000

[0127]

[0128] The formulation is administered via subcutaneous injection prenatally or perinatally and is composed of genetic cargo encapsulated in LNPs that are suspended in PBS .

[0129] Example 3

[0130] A lipid nanoparticle formulation is prepared with an N to P ratio of 7 and contains a therapeutic transgene (adalimumab) for the treatment of rheumatoid arthritis . The LNPs are targeted to adipocytes using a conjugated antibody against aquaporin 7B . The antibody is conjugated to the LNPs using a thiol-maleimide reaction with N-hydroxysuccinamide from the DOPE-NHS .

[0131] Molar Lipid Formal Name

[0132] ratio, % 8- [ ( 2-hydroxyethyl) [ 6-oxo-6- SM-102 (undecyloxy) hexyl] amino] -octanoic acid, 1- 50

[0133] octylnonyl ester

[0134] N- ( Succinimidyloxy-glutaryl) -L-a- DOPE-NHS 10

[0135] phosphat idyl ethanolamine, Dioleoyl

[0136] Cholesterol (3p) -cholest-5-en-3-ol 38 .5

[0137] 1, 2-dimyristoyl-rac-glycero-3- DMG-PEG2000 1. 5

[0138] methoxypolyethylene glycol-2000

[0139]

[0140] The formulation is administered via subcutaneous or intravenous injection and is composed of genetic cargo encapsulated in anti-aquaporin 7B-con jugated LNPs that are suspended in PBS . Following administration via a subcutaneous injection the formulation remains highly localized to the adipose tissues with over 99% localization . Following administration via an intravenous injection, the formulation localizes to the most proximal adipose tissues downstream of the injection, and at least in part to the downstream tissues and organs, with some localization in the liver .

[0141] Example 4 A lipid nanoparticle formulation is prepared with an N to P ratio of 7 , and contains a therapeutic transgene (beta-glucocerebrosidase) for the treatment of Gaucher' s Disease .

[0142] Molar Lipid Formal Name

[0143] ratio, % 6- ( ( 2-hexyldecanoyl) oxy) -N- ( 6- ( ( 2- ALC-0315 hexyldecanoyl) oxy) hexyl) -N- ( 4- 46.3 hydroxybutyl) hexan-l-aminium

[0144] DSPC 1, 2-distearoyl-sn-glycero-3-phosphocholine 9. 4

[0145] Cholesterol (3p) -cholest-5-en-3-ol 42 . 7

[0146] 1, 2-dimyristoyl-rac-glycero-3- DMG-PEG2000 1. 6 methoxypolyethylene glycol-2000

[0147]

[0148] The formulation is administered topically as a petrolatum-based ointment and is composed of genetic cargo encapsulated in LNPs that are suspended in petroleum jelly .

[0149] Example 5

[0150] A lipid nanoparticle formulation is prepared with an N to P ratio of 7 and contains a therapeutic transgene (alpha-glucosidase) for the treatment of Pompe Disease .

[0151] Molar Lipid Formal Name

[0152] ratio, % Coatsome

[0153] n / a 48 . 5 SS-OP

[0154] DOPC 1, 2-dioleoyl-sn-glycero-3-phosphocholine 7 . 5 Cholesterol (3p) -cholest-5-en-3-ol 42 . 5

[0155] 1, 2-dimyristoyl-rac-glycero-3- DMG-PEG2000 1 . 5 methoxypolyethylene glycol-2000

[0156]

[0157] The formulation is administered via a subcutaneous or intravenous injection and is composed of genetic cargo encapsulated in LNBs that are suspended in PBS . The subcutaneous formulation can be administered as a ImL injection and re-dosed as required, at a frequency of approximately once per week or once per month to obtain optimal therapeutic effect .

[0158] Example 6

[0159] Adipocyte targeting compositions and ligands can enhance localization of the genetic cassette in the subcutis and specifically within subcutaneous adipose tissues or white adipocytes . Such ligands and compositions can afford homogeneous transfection of adipocytes without transfection of neighboring cells and structures such as hair follicles, blood vessels, epidermis, dermis, and muscles . Such compositions also benefit from the enhanced localization, minimizing off-target DNA or RNA delivery, reduce liver impact of treatment, and enable transfection of a durable secretory cell type with easy access via subcutaneous delivery.

[0160] Adipocyte targeting compositions and targeting ligands were used to enhance localization of the treatment to subcutaneous adipose tissues, which was confirmed by reporter gene IHC staining. Briefly, white subcutaneous adipocytes were homogeneously transfected with said formulations, while neighboring tissues such as blood vessels and hair follicles remained free of the delivered transgene (as assessed by reported gene IHC staining for an intracellular reporter protein) .

[0161] Brown staining indicates positive staining, blue / pink / light staining indicates absence of positive staining for the intracellular reporter protein .

[0162] Adipocyte targeted vectors resulted in durable expression for the lifetime of adipocytes, which in humans is estimated at approximately 10 years . Adipocyte targeted non-viral vectors produced durable expression over a period of 12 months with no measurable decline in expression levels . In contrast, liver-targeted, systemically administered, or muscle-targeted vectors display a clear spike-decline type kinetics, where expression initially rapidly increased to form a spike, which was followed by rapid decline (often 1 or more orders of magnitude) , followed by eventual perpetual decay . Such pharmacokinetics are not optimal for the overwhelming majority of delivered biologies, therefore markedly increasing utility of adipocyte targeted vectors .

[0163] Biodistrubtion of the adipocyte targeted vectors was additionally confirmed at the 12-month study timepoint by ex vivo imaging of explanted tissues from C57BL / 6 mice . Confirming the immunohistochemical findings the vectors were predominantly localized to the subcutis, with only minor distal tissue biodistribution, which may have occurred due to broken blood vessels during injection .

Claims

CLAIMSWhat is claimed is :1 . A nanoparticle formulation for the delivery of genetic cargo to subcutaneous adipose tissues , which includes :a . At least one ioni zable lipid, or lipidoid, or lipid-like molecule, or intracellular vesicle-interacting peptide, and b . A structural element comprised of at least one polymer or structural and cationic lipid, andc . Optionally at least one cholesterol or cholesterol analog or cholesterol substitute , andd . Optionally a targeting element , ande . A genetic cargo of 1 . 5-7 . 0kb in length encoding at least one therapeutic gene and formulated in an N to P ratio of 3 . 5 to 9 . 5 .2 . An ioni zable lipid in Claim 1 , which is a cationic lipid .3 . An ioni zable lipid in Claim 1 , which is a dimethylammonium-propane or analog .4 . An ioni zable lipid in Claim 1 , which is 18 : 0 DAP ( 1 , 2-distearoyl- 3-dimethylammonium-propane ) or analog .5 . An ionizable lipid in Claim 1 , which is 16 : 0 DAP ( 1 , 2-dipalmitoyl- 3-dimethylammonium-propane ) or analog .6 . An ionizable lipid in Claim 1 , which is 14 : 0 DAP ( 1 , 2-dimyristoyl- 3-dimethylammonium-propane ) or analog .7 . An ionizable lipid in Claim 1 , which is 18 : 1 DAP or DODAP ( 1 , 2 — dioleoyl-3-dimethylammonium-propane) or analog .8 . An ioni zable lipid in Claim 1 , which is Lipid C24 (bis ( 2- octyldodecyl) 3 , 3 ' - ( ( 4- ( 4-methylpiperazin-l- yl) butyl) azanediyl) dipropionate) or analog .9 . An ionizable lipid in Claim 1 , which is ATX-001 ( 8 , 8f— [ [ 2— [ [ 2— ( dimethylamino) ethyl ] thio ] acetyl ] imino ] bis-octanoic acid, 1 , 1 ' - di- ( 2 Z ) -2-nonen-l-yl ester) or analog .An ionizable lipid m Claim 1, which is Lipid A9 (bus (2-butyloctyl) 10- (N- (3- (dimethylamino) propyl) nonanamido) nonadecanedioate) or analog .An ionizable lipid in Claim 1, which is CIN-16645 ( 9Z, 12Z-octadecadienoic acid, 3- [4 , 4-bis (octyloxy) -1-oxobutoxy] -2- [ [ [ [3- ( diethylamino) propoxy] carbonyl] oxy] methyl] propyl ester) or analog.An ionizable lipid in Claim 1, which is Lipid 23 ( 8— ( (2— ( ( 4 — (dimethylamino) butanoyl) oxy) ethyl) (10-oxo-10- (tridecan-7-yloxy) decyl) amino) octyl 2-hexyldecanoate) or analog.An ionizable lipid in Claim 1, which is Lipid 29 ( 8— [ [8— [ ( 1— ethylnonyl) oxy] -8-oxooctyl] [3- [ [2- (methylamino) -3, 4-dioxo-l-cyclobuten-l-yl ] amino ] propyl ] amino ] -octanoic acid, 1-octylnonyl ester) or analog.An ionizable lipid in Claim 1, which is Lipid 8 (4- (dimethylamino) -butanoic acid, 2- [di- ( 9Z, 12Z) -9, 12-octadecadien-1-ylamino ] ethyl ester) or analog .An ionizable lipid in Claim 1, which is 1014 (15, 19, 28-tris (2-hydroxytetradecyl) -15, 19, 24 , 28-tetraazadotetracontane-13, 30-diol) or analog .An ionizable lipid in Claim 1, which 246C10 ( 1 , 1 ' , 1 ' ' , 1 ' ' ' - [ 1, 4-piperazinediylbis (3, 1-propanediylnitrilo) ] tetrakis-2-dodecanol) or analog.An ionizable lipid in Claim 1, which is 98N12-5 (N1, N16-didodecyl-4 , 7 , 13-tris [3- (dodecylamino) -3-oxopropyl ] -4 , 7 , 10, 13-tetraazahexadecanediamide) or analog .An ionizable lipid in Claim 1, which is YK-009 ( 6— [ [ 4— (decyloxy) -4-oxobutyl] (2-hydroxyethyl) amino] -hexanoic acid, 2-octyldecyl ester) or analog.An ionizable lipid in Claim 1, which is CL4F8-6 ( 7- ( 4- ( dipropylamino) butyl) -7-hydroxytridecane-l , 13-diyl bis (2-hexyloctanoate) ) or analog .An ionizable lipid in Claim 1, which is IC8 ( 1 , 11, 11 1, 11 1 1- ( (piperazine-1, 4-diylbis (propane-3, 1-diyl) ) bis (azanetriyl) ) tetrakis (tetradecan-2-ol) ) or analog.An ionizable lipid m Claim 1 , which is Lipid I II-45 ( 2-butyl-octanoic acid, 1 , 1 ' - [ [ ( 3-hydroxypropyl) imino ] di-9 , 1-nonanediyl ] ester) or analog .An ionizable lipid in Claim 1 , which is ATX-100 ( 4 , 4 ' - [ [ [ [ 3- ( dimethylamino ) propyl ] thio ] carbonyl ] imino ] bis-butanoic acid, 1 , 1 ' -bis ( 1-heptyloctyl ) ester) or analog .An ionizable lipid in Claim 1 , which is D0G-IM4 (N— [ ( 25 Z ) — 14- [ ( 9Z ) -9-octadecen-l-yloxy ] -3 , 6, 9, 12 , 16-pentaoxatetratriacont-25-en-l-yl ] -lH-imidazole-5-carboxamide ) or analog .An ioni zable lipid in Claim 1 , which is OF-Deg-Lin ( 9Z , 12 Z-octadecadienoic acid, 1 , 1 ' , 1 ' ' , 1 ' ' ' - [ ( 3 , 6-dioxo-2 , 5-piperazinediyl ) bis ( 4 , l-butanediylnitrilodi-2 , 1-ethanediyl ) ] ester) or analog .An ionizable lipid in Claim 1 , which is 0F-C4-Deg-Lin ( 9, 12-octadecadienoic acid, ( 9Z , 12 Z ) -1 , 1 ' , 1 ' ' , 1 ' ' ' - [ ( 3 , 6-dioxo-2 , 5-piperazinediyl) bis ( 4 , l-butanediylnitrilodi-4 , 1-butanediyl ) ] ester) or analog .An ionizable lipid in Claim 1 , which is ATX-0114 (heptadecan-9-yl ( Z ) -N- ( ( ( 4- (dimethylamino ) butyl) thio ) carbonyl) -N- ( 2- ( non- 2-en-l-yloxy) -2-oxoethyl) glycinate) or analog .An ioni zable lipid in Claim 1 , which is L202 ( l-methyl-4-piperidinecarboxylic acid, 2- [ 9-oxo-9- [ ( 3-pentyloctyl ) oxy] nonyl ] dodecyl ester) or analog .An ionizable lipid in Claim 1 , which is C13-112-tetra-tail ( 14 , 23-bis ( 2-hydroxytridecyl) -17 , 20-dioxa-14 , 23-diazahexatriacontane-12 , 25-diol ) or analog .An ionizable lipid in Claim 1 , which is OC2-K3-E10 ( 3 , 3 ' - ( ( 2-hydroxyethyl) azanediyl) bis (N- ( 3- (bis ( 2-hydroxydecyl) amino) propyl ) propanamide ) or analog .An ionizable lipid in Claim 1 , which is RCB-4-8 (bis ( ( Z ) -12- ( ( ( non-2-yn-l-yloxy) carbonyl) oxy) octadec-9-en-l-yl ) 3 , 3 ' - ( ( 2- ( diethylamino ) ethyl ) azanediyl ) dipropionate) or analog .An ionizable lipid in Claim 1 , which is RM 133-3 ( ( 9Z , 9 ' Z ) - ( (3 , 3 ’ - ( ( 3- ( diethylamino ) propyl) azanediyl) bis (propanoyl) ) bis ( oxy) ) bis ( octadec-9-ene-18 , 7-diyl) ( 5 Z , 5 ' Z , 8 Z , 8 ' Z , 11Z , 11 ' Z , 14 Z , 14,Z , 17 Z , 17,Z ) -bis ( icosa-5 , 8 , 11 , 14 , 17-pentaenoate ) ) or analog .An ioni zable lipid in Claim 1 , which is RM 137-5 ( ( 9Z , 9 ' Z ) - ( ( 3 , 3 ' - ( ( 2- ( 4-methylpiperazin-l-yl ) ethyl ) azanediyl ) bis (propanoyl ) ) bis ( oxy) ) bis (octadec-9-ene-18 , 7-diyl) ( 4 Z , 4 ' Z , 7 Z , 7 ' Z , 10 Z , 10 ' Z , 13Z , 13 ' Z , 16Z , 16 ' Z , 19Z , 19 ' Z ) -bis ( docosa-4 , 7 , 10 , 13 , 16, 19-hexaenoate) ) or analog .An ionizable lipid in Claim 1 , which is PP Z-A10 (N, N ' - (piperazine-1, 4-diylbis (propane-3 , 1-diyl ) ) bis ( 3- (didecylamino) propanamide) ) or analog .An ioni zable lipid in Claim 1 , which is Lipid 10 ( 4-methyl- 1-piperazinepropanoic acid, 2- [di- ( 9Z , 12 Z ) -9 , 12-octadecadien-l-ylamino ] ethyl ester) or analog .An ionizable lipid in Claim 1 , which is Lipid AX4 or analog . An ioni zable lipid in Claim 1 , which is Lipid 5 ( 8— [ ( 2— hydroxyethyl ) [ 8- (nonyloxy) -8-oxooctyl ] amino ] -octanoic acid, 1-octylnonyl ester) or analog .An ionizable lipid in Claim 1 , which is TCL053 ( 2- ( ( ( 4- (dimethylamino ) butanoyl) oxy) methyl ) -2- ( ( ( ( Z ) -tetradec-9-enoyl) oxy) methyl ) propane-1 , 3-diyl ( 9Z , 9 ' Z ) -bis (tetradec-9-enoate ) ) or analog .An ioni zable lipid in Claim 1 , which is YSK05 ( l-methyl-4 , 4-bis [ ( 9Z , 12 Z ) -9 , 12-octadecadien-l-yloxy] -piperidine ) or analog .An ionizable lipid in Claim 1 , which is L-319 ( 9— [ 4— (dimethylamino ) -1-oxobutoxy] -heptadecanedioic acid, 1 , 17-di- ( 2 Z ) - 2-nonen-l-yl ester) or analog .An ionizable lipid in Claim 1 , which is 4A3-SC8 or analog . An ionizable lipid in Claim 1 , which is C14-4 ( 1 , 1 [ [ 2- [ 2- [ 4- [ 2- [ [ 2- [ 2- [bis ( 2-hydroxytetradecyl) amino ] ethoxy] ethyl ] ( 2-hydroxytetradecyl) amino ] ethyl ] -1-piperazinyl ] ethoxy] ethyl ] imino ] bis-2-tetradecanol) or analog . An ioni zable lipid in Claim 1 , which is DODMA ( 1 , 2-dioleyloxy- 3-dimethylaminopropane ) or analog .An ionizable lipid m Claim 1, which is DOBAQ (N— ( 4— carboxybenzyl) -N, N-dimethyl-2 , 3-bis (oleoyloxy) propan-l-aminium) or analog.An ionizable lipid in Claim 1, which is a cationic amino lipid.An ionizable lipid in Claim 1, which is DLin-DMA (N, N-dimethyl-2 , 3-bis [ ( 9Z , 12Z ) -9, 12-octadecadien-l-yloxy] -1-propanamine) or analog .An ionizable lipid in Claim 1, which is TT3 (Nl, N3 , N5-tris [ 3- (didodecylamino) propyl] -1, 3, 5-benzenetricarboxamide) or analog .An ionizable lipid in Claim 1, which is FTT5 (Hexa (octan-3-yl) 9, 9 ' , 9", 9"', 9"", 9"'"- ( ( ( (benzene-1 , 3 , 5-tricarbonyl) ris (azanediyl) ) tris (propane-3, 1-diyl) ) tris (azanetriyl) ) hexanonanoate) .An ionizable lipid in Claim 1, which is OF-02 (3, 6-bis [ 4- [bis [ ( 9Z, 12Z) -2 -hydroxy- 9, 12 -octade cadie n- 1 -y 1 ] amino] butyl] -2, 5-piperazinedione) or analog .An ionizable lipid in Claim 1, which is AA3-DLin (piperazine-1, 4-diylbis ( ethane -2 , 1-diyl) ( 9 Z , 91Z, 12 Z, 121Z) -bis (octadeca- 9, 12-dienoate) ) or analog .An ionizable lipid in Claim 1, which is DLin-MC3-DMA ( 4- (dimethylamino) -butanoic acid, ( 10 Z, 13Z) -1- ( 9Z , 12Z ) -9, 12-octadecadien-l-yl-10 , 13-nonadecadien-l-yl ester) or analog.An ionizable lipid in Claim 1, which is DLin-KC2-DMA (N, N-dimethyl-2 , 2 -di- ( 9Z, 12Z) -9, 12-octadecadien-l-yl-l, 3-dioxolane-4-ethanamine) or analog.An ionizable lipid in Claim 1, which is SM-102 ( 8— [ (2— hydroxyethyl) [ 6-oxo-6- (undecyloxy) hexyl] amino] -octanoic acid, 1-octylnonyl ester) or analog.An ionizable lipid in Claim 1, which is ALC-0315 ( 6- ( (2-hexyldecanoyl) oxy) -N- ( 6- ( ( 2-hexyldecanoyl) oxy) hexyl) -N- ( 4-hydroxybutyl) hexan-l-aminium) or analog.An ionizable lipid in Claim 1, which is ALC-0315 analogous-1 ( ( (2-hydroxyethyl) azanediyl) bis (octane-8 , 1-diyl) bis (2-hexyldecanoate) ) or analog .An ionizable lipid m Claim 1, which is Lipid 222 ( ( (4-hydroxybutyl) azanediyl) bis (octane-8, 1-diyl) bis (2-hexyldecanoate) ) or analog .An ionizable lipid in Claim 1, which is CL4-H6 ( 9-octadecenoic acid, 1, 1 [ 7- [4- (dipropylamino) butyl] -7-hydroxy-l, 13-tridecanediyl] ester) or analog.An ionizable lipid in Claim 1, which is AL-A12 (N-dodecyl-N- (3-methoxypropyl) dodecan-l-amine) or analog.An ionizable lipid in Claim 1, which is Lipid 14 ( ( (2- ( ( 4- ( dimethylamino) butanoyl) oxy) ethyl) azanediyl) bis (octane-8, 1-diyl) bis (2-hexyldecanoate) ) or analog .An ionizable lipid in Claim 1, which is Lipid 16 ( ( (2- ( ( 4- (dimethylamino) butanoyl) oxy) ethyl) azanediyl) bis (octane-8 , 1-diyl) bis (2-hexyldecanoate) ) or analog .An ionizable lipid in Claim 1, which is a cationic alkyne lipid.An ionizable lipid in Claim 1, which is Lipid A6 (di (dec-3-yn-l-yl) 9- ( (4- (dimethylamino) butanoyl) oxy) heptadecanedioate) or analog .An ionizable lipid in Claim 1, which is a cationic trialkyl lipid.An ionizable lipid in Claim 1, which is Lipid CL1 (5- (dimethylamino) -pentanoic acid, ( 6Z) -1, 2-di- ( 4 Z ) -4-decen-l-yl-6-dodecen-l-yl ester) or analog.An ionizable lipid in Claim 1, which is a disulfide-bond containing cationic lipid.An ionizable lipid in Claim 1, which is a BAMEA-016B (bis (2- ( dodecyldisulf aneyl) ethyl) 3, 3 ' - ( (3-methyl-9-oxo-10-oxa-13, 14-dithia-3, 6-diazahexacosyl) azanediyl) dipropionate) or analog.An ionizable lipid in Claim 1, which is Coatsome SS-E (Bis { 2- [N-methyl-N- (a-D-tocopherolhemisuccinatepropyl) amino] ethyl } disulf ide) or analog .An ionizable lipid in Claim 1, which is Coatsome SS-EC (Bis { 2- [4- (a-D-tocopherolhemisuccinateethyl) piperidyl] ethyl } disulfide) or analog.An ionizable lipid m Claim 1, which is Coatsome SS-OP (is Coatsome SS-OC (Bis { 2- [4- (cis-9-octadecenoateethyl) -1-piperidinyl] ethyl } disulf ide) or analog .An ionizable lipid in Claim 1, which is Coatsome SS-OP (Bis [2- ( 4- { 2- [ 4- (cis-9-octadecenoyloxy) phenylacetoxy] ethyl }piperidinyl) ethyl] disulfide) or analog .An ionizable lipid in Claim 1, which is a lipomer .An ionizable lipid in Claim 1, which is CKK-E12 (3, 6-bis [4- [bis (2-hydroxydodecyl) amino] butyl] -2 , 5-piperazinedione) or analog .An ionizable lipid in Claim 1, which is a lipidoid or lipidoid analog .An ionizable lipid in Claim 1, which is C12-113 ( 1, 1 ' , 1 ' ' , 1 ' ' '- [ (methylimino) bis (2, 1-ethanediylnitrilo) ] tetrakis-2-dodecanol) or analog.An ionizable lipid in Claim 1, which is C12-200 ( 1, 1 ' - [ [2— [ 4- [2- [ [2- [bis (2-hydroxydodecyl) amino] ethyl] (2-hydroxydodecyl) amino] ethyl] -1-piperazinyl] ethyl] imino] bis-2-dodecanol) or analog.An ionizable lipid in Claim 1, which is NT1-014B or analog. An ionizable lipid in Claim 1, which is 306-O12B-3 (bis (2- (octyldisulfaneyl) ethyl) 3, 31- ( ( 4-methyl-ll-oxo-12-oxa-15 , 16-dithia-4 , 8-diazatetracosyl) azanediyl) dipropionate) or analog.An ionizable lipid in Claim 1, which is 503-013 or analog . An ionizable lipid in Claim 1, which is a disulfide-cleavable proton-activated lipid like material .An ionizable lipid in Claim 1, which is SS-Palm-M (tetradecanoic acid, 1, 1 ' - [dithiobis [2, 1-ethanediyl (methylimino) -3 , 1-propanediyl ] ] ester) or analog.An ionizable lipid in Claim 1, which is SS-PalmO-Phe (4- [ [ ( 9Z ) -l-oxo-9-octadecen-l-yl ] oxy ] -benzeneacetic acid, 1, 1 ' - [dithiobis (2, 1-ethanediyl-l , 4-piperidinediyl-2 , 1-ethanediyl) ] ester) or analog.A structural Irprd in Clarm 1, whrch is a saturated phosphatidic acid.A structural lipid in Claim 1, which is DMPA ( 1 , 2-dimyristoyl-sn-glycero-3-phosphate) or analog.A structural lipid in Claim 1, which is DPPA ( 1 , 2-dipalmitoyl-sn-glycero-3-phosphate) or analog.A structural lipid in Claim 1, which is DSPA ( 1, 2-distearoyl-sn-glycero-3-phosphate) or analog.A structural lipid in Claim 1, which is DLPA ( 1 , 2-dilauroyl-sn-glycero-3-phosphate) or analog.A structural lipid in Claim 1, which is an unsaturated phosphatidic acid.A structural lipid in Claim 1, which is DOPA ( 1, 2-dioleoyl-sn-glycero-3-phosphate) or analog.A structural lipid in Claim 1, which is DAPA ( 1, 2-diarachidonoyl-sn-glycero-3-phosphate) or analog .A structural lipid in Claim 1, which is a phosphatidic acid with both saturated and unsaturated tails .A structural lipid in Claim 1, which is POPA ( l-palmitoyl-2-oleoyl-sn-glycero-3-phosphate) or analog .A structural lipid in Claim 1, which is PAPA ( l-palmitoyl-2-arachidonoyl-sn-glycero-3-phosphate) or analog .A structural lipid in Claim 1, which is SOPAPA ( 1-stearoyl-2-oleoyl-sn-glycero-3-phosphate) or analog .A structural lipid in Claim 1, which is SAPA ( l-stearoyl-2-arachidonoyl-sn-glycero-3-phosphate) or analog.A structural lipid in Claim 1, which is a saturated phosphatidylcholine .A structural lipid in Claim 1 which is DHPC ( 1, 2-dihexanoyl-sn-glycero-3-phosphocholine or 1 , 2-diheptanoyl-sn-glycero-3-phosphocholine) or analog .A structural lipid in Claim 1, which is DLPC ( 1, 2-dilauroyl-sn-glycero-3-phosphocholine) or analog.A structural lipid in Claim 1, which is DMPC ( 1, 2-dimyristoyl-sn-glycero-3-phosphocholine) or analog.98 . A structural lipid in Claim 1, which is DPPC ( 1 , 2-dipalmitoyl- sn-glycero-3-phosphocholine) or analog .

99. A structural lipid in Claim 1, which is DSPC ( 1, 2-distearoyl- sn-glycero-3-phosphocholine) or analog .100 . A structural lipid in Claim 1, which is an unsaturated phosphatidylcholine .

101. A structural lipid in Claim 1, which is DOPC ( 1, 2-dioleoyl- sn-glycero-3-phosphocholine) or analog .102 . A structural lipid in Claim 1, which is DLPC ( 1 , 2-dilinoleoyl- sn-glycero-3-phosphocholine) or analog .

103. A structural lipid in Claim 1, which is DEPC ( 1 , 2-dierucoyl- sn-glycero-3-phosphocholine) or analog .104 . A structural lipid in Claim 1, which is 1, 2-DAPC (1, 2- Diarachidoyl-sn-glycero-3-phosphocholine) or analog.

105. A structural lipid in Claim 1, which is DDPC ( 1, 2-Didecanoyl- sn-glycero-3-phosphocholine) or analog .

106. A structural lipid in Claim 1, which is 1, 2-PLPC ( 1-Palmitoyl- 2-Lauroyl-sn-glycero-3-Phosphocholine) or analog .107 . A structural lipid in Claim 1, which is an unsaturated phosphatidylcholine with tails of different numbers of carbon atoms .108 . A structural lipid in Claim 1, which is MPPC ( l-myristoyl-2- palmitoyl-sn-glycero-3-phosphocholine) or analog .

109. A structural lipid in Claim 1, which is MSPC ( l-myristoyl-2- stearoyl-sn-glycero-3-phosphocholine) or analog .110 . A structural lipid in Claim 1, which is PMPC ( l-palmitoyl-2- myristoyl-sn-glycero-3-phosphocholine) or analog.

111. A structural lipid in Claim 1, which is PSPC ( l-palmitoyl-2- stearoyl-sn-glycero-3-phosphocholine) or analog .112 . A structural lipid in Claim 1, which is SMPC ( l-stearoyl-2- myristoyl-sn-glycero-3-phosphocholine) or analog.

113. A structural lipid in Claim 1, which is SPPC ( l-stearoyl-2- palmitoyl-sn-glycero-3-phosphocholine) or analog.114 . A structural lipid in Claim 1, which is a phosphatidylcholine with both saturated and unsaturated tails .

115. A structural lipid in Claim 1, which is POPC ( l-palmitoyl-2- oleoyl-glycero-3-phosphocholine) or analog .

116. A structural lipid in Claim 1, which is PAPC ( l-palmitoyl-2- arachidonoyl-sn-glycero-3-phosphocholine) or analog.117 . A structural lipid in Claim 1, which is SOPC ( l-stearoyl-2- oleoyl-sn-glycero-3-phosphocholine) or analog.118 . A structural lipid in Claim 1, which is OMPC (l-oleoyl-2- myristoyl-sn-glycero-3-phosphocholine) or analog .

119. A structural lipid in Claim 1, which is OPPC (l-oleoyl-2- palmitoyl-sn-glycero-3-phosphocholine) or analog .120 . A structural lipid Claim 1, which is OSPC ( l-oleoyl-2- stearoyl-sn-glycero-3-phosphocholine) or analog.

121. A structural lipid in Claim 1, which is a lysophosphatidylcholine .122 . A structural lipid in Claim 1, which is S-LysoPC ( 1-Stearoyl- 2-lyso-sn-glycero-3-phosphocholine) or analog.

123. A structural lipid in Claim 1, which is P-LysoPC ( 1-Palmitoyl- 2-lyso-sn-glycero-3-phosphocholine) or analog.124 . A structural lipid in Claim 1, which is O-LysoPC ( l-01eoyl- 2-lyso-sn-glycero-3-phosphocholine) or analog.

125. A structural lipid in Claim 1, which is M-LysoPC ( 1-Myristoyl- 2-lyso-sn-glycero-3-phosphocholine) or analog.

126. A structural lipid in Claim 1, which is a saturated phosphatidylethanolamine .127 . A structural lipid in Claim 1, which is DLPE ( 1 , 2-dilauroyl- sn-glycero-3-phosphoethanolamine) or analog.128 . A structural lipid in Claim 1, which is DPPE ( 1 , 2-dipalmitoyl- sn-glycero-3-phosphoethanolamine) or analog.

129. A structural lipid in Claim 1, which is DPPE-Mal (N- (3- Maleimide-1 -oxopropyl) -L-a-phosphat idyl ethanolamine, Dipalmitoyl) or analog .130 . A structural lipid in Claim 1, which is DPPE-NHS (N- ( Succinimidyloxy-glutaryl) -L-a-phosphat idyl ethanolamine, Dipalmitoyl) or analog .

131. A structural lipid in Claim 1, which is DPPE-Glu (N-Glutaryl- L-a-phosphatidylethanolamine, Dipalmitoyl) or analog .132 . A structural lipid in Claim 1, which is DSPE ( 1, 2-distearoyl- sn-glycero-3-phosphoethanolamine) or analog.

133. A structural lipid in Claim 1, which is DSPE-Mal (N- (3- Mal eimi de- 1 -oxopropyl) -L-a-phosphatidy let hanol amine, Di st earoyl) or analog .134 . A structural lipid in Claim 1, which is DSPE-NHS (N- ( Succinimidyloxy-glutaryl) -L-a-phosphatidy let hanol amine, Distearoyl) or analog.

135. A structural lipid in Claim 1, which is DSPE-Glu (N-Glutaryl- L-a-phosphatidylethanolamine, Distearoyl) or analog.

136. A structural lipid in Claim 1, which is DMPE ( 1 , 2-dimyristoyl- sn-glycero-3-phosphoethanolamine) or analog.

137. A structural lipid in Claim 1, which is DMPE-Mal (N- (3- Male imide- 1 -oxopropyl) -L-a-phosphatidy let hanol amine, Dimyristoyl) or analog.

138. A structural lipid in Claim 1, which is DMPE-NHS (N- ( Succinimidyloxy-glutaryl) -L-a-phosphatidy let hanol amine, Dimyristoyl) or analog .

139. A structural lipid in Claim 1, which is DMPE-Glu (N-Glutaryl- L-a-phosphatidylethanolamine, Dimyristoyl) or analog .

140. A structural lipid in Claim 1, which is an unsaturated phosphatidylethanolamine .

141. A structural lipid in Claim 1, which is DOPE ( 1, 2-dioleoyl- sn-glycero-3-phosphoethanolamine) or analog.

142. A structural lipid in Claim 1, which is DOPE-Mal (N- (3- Maleimide-l-oxopropyl) -L-a-phosphatidylethanolamine, Dioleoyl) or analog .

143. A structural lipid in Claim 1, which is DOPE-NHS (N- ( Succinimidyloxy-glutaryl) -L-a-phosphatidy let hanol amine, Dioleoyl) or analog.

144. A structural lipid in Claim 1, which is DOPE-Glu (N-Glutaryl- L-a-phosphatidylethanolamine, Dioleoyl) or analog.

145. A structural lipid in Claim 1, which is DAPE ( 1, 2- diarachidonoyl-sn-glycero-3-phosphoethanolamine) or analog.

146. A structural lipid in Claim 1, which is a phosphatidylethanolamine with both saturated and unsaturated tails .147 . A structural lipid in Claim 1, which is POPE ( l-palmitoyl-2- oleoyl-sn-glycero-3-phosphoethanolamine) or analog .148 . A structural lipid in Claim 1, which is SOPE ( l-stearoyl-2- oleoyl-sn-glycero-3-phosphoethanolamine) or analog .

149. A structural lipid in Claim 1, which is PAPE ( l-palmitoyl-2- arachidonoyl-sn-glycero-3-phosphoethanolamine) or analog.150 . A structural lipid in Claim 1, which is SAPE ( l-stearoyl-2- arachidonoyl-sn-glycero-3-phosphoethanolamine) or analog.

151. A structural lipid in Claim 1, which is a saturated phosphatidylglycerol .152 . A structural lipid in Claim 1, which is DLPG ( 1 , 2-dilauroyl- sn-glycero-3-phospho- (1 ' -rac-glycerol) ) or analog.

153. A structural lipid in Claim 1, which is DMPG ( 1, 2-dimyristoyl- sn-glycero-3-phospho- (1 ' -rac-glycerol) ) or analog .154 . A structural lipid in Claim 1, which is DPPG ( 1, 2-dipalmitoyl- sn-glycero-3-phospho- (1 ' -rac-glycerol) ) or analog .

155. A structural lipid in Claim 1, which is DSPG ( 1 , 2-distearoyl- sn-glycero-3-phospho- (1 ' -rac-glycerol) ) or analog .

156. A structural lipid in Claim 1, which is DOPG ( 1 , 2-dioleoyl- sn-glycero-3-phospho- (1 ' -rac-glycerol) ) or analog .157 . A structural lipid in Claim 1, which is an unsaturated phosphatidylglycerol .158 . A structural lipid in Claim 1, which is a phosphatidylglycerol with both saturated and unsaturated tails .

159. A structural lipid in Claim 1, which is POPG ( l-palmitoyl-2- oleoyl-sn-glycero-3-phospho- ( 1 ' -rac-glycerol) ) or analog.160 . A structural lipid in Claim 1, which is PAPG ( l-palmitoyl-2- arachidonoyl-sn-glycero-3-phospho- ( 1 ' -rac-glycerol) ) or analog.

161. A structural lipid in Claim 1, which is SOPG (l-stearoyl-2- oleoyl-sn-glycero-3-phospho- ( 1 ' -rac-glycerol) ) or analog.

162. A structural lipid in Claim 1, which is SAPG ( l-stearoyl-2- arachidonoyl-sn-glycero-3-phospho- (1 ' -rac-glycerol) ) or analog.

163. A structural lipid in Claim 1, which is a saturated phosphatidylserine .164 . A structural lipid in Claim 1, which is DSPS ( 1, 2-distearoyl- sn-glycero-3-phospho-L-serine) or analog .

165. A structural lipid in Claim 1, which is DMPS ( 1 , 2-dimyristoyl- sn-glycero-3-phospho-L-serine) or analog .

166. A structural lipid in Claim 1, which is DPPS ( 1 , 2-dipalmitoyl- sn-glycero-3-phospho-L-serine) or analog .167 . A structural lipid in Claim 1, which is DLPS ( 1 , 2-dilauroyl- sn-glycero-3-phospho-L-serine) or analog .168 . A structural lipid in Claim 1, which is an unsaturated phosphatidylserine .

169. A structural lipid in Claim 1, which is DOPS ( 1, 2-dioleoyl- sn-glycero-3-phospho-L-serine) or analog .170 . A structural lipid in Claim 1, which is a phosphatidylserine with both saturated and unsaturated tails .

171. A structural lipid in Claim 1, which is POPS ( l-palmitoyl-2- oleoyl-sn-glycero-3-phospho-L-serine) or analog .172 . A structural lipid in Claim 1, which is SOPS ( l-stearoyl-2- oleoyl-sn-glycero-3-phospho-L-serine) or analog .

173. A structural lipid in Claim 1, which is PAPS ( l-palmitoyl-2- arachidonoyl-sn-glycero-3-phospho-L-serine) or analog .174 . A structural lipid in Claim 1, which is SAPS ( l-stearoyl-2- arachidonoyl-sn-glycero-3-phospho-L-serine) or analog .

175. A cationic lipid in Claim 1, which is DORI (N— (2 — hydroxyethyl) -N, N-dimethyl-2 , 3-bis (oleoyloxy) propan-l-aminium bromide) or analog.

176. A cationic lipid in Claim 1, which is DC-6-14 (0, 0' - ditetradecanoyl-N- (a-trimethylammonioacetyl) diethanolamine) or analog .177 . A cationic lipid in Claim 1, which is DSDMA ( 1, 2-distearyloxy- N, N-dimethyl-3-aminopropane) or analog.

178. A cationic lipid in Claim 1, which is DMRIE (1, 2- dimyristyloxypropyl-3-dimethyl-hydroxyethyl ammonium bromide) or analog .

179. A cationic lipid in Claim 1, which is a quaternary ammonium compound .

180. A cationic lipid in Claim 1, which is 18 : 0 DDAB (Dimethyldioctadecylammonium) or analog.

181. A cationic lipid in Claim 1, which is DC-1-16 (N-Hexadecyl- N, N-dimethylhexadecan-l-aminium) or analog .

182. A cationic lipid in Claim 1, which is dimethyldimyristylammonium (N, N-dimethyl-N-tetradecyl-1- tetradecanaminium) or analog .

183. A cationic lipid in Claim 1, which is DODAC N, N-Dioleyl-N, N- dimethylammonium chloride) or analog .184 . A cationic lipid in Claim 1, which is a trimethylammoniumpropane .

185. A cationic lipid in Claim 1, which is 14 : 0 TAP ( 1, 2- dimyristoyl-3-trimethylammoniumpropane) or analog.

186. A cationic lipid in Claim 1, which is 16 : 0 TAP ( 1, 2 — dipalmitoyl-3-trimethylammoniumpropane) or analog.

187. A cationic lipid in Claim 1, which is 18 : 0 TAP ( 1, 2-stearoyl- 3-trimethylammoniumpropane) or analog.

188. A cationic lipid in Claim 1, which is 18 : 1 TAP (DOTAP, 1, 2- dioleoyl-3-trimethylammoniumpropane) or analog .

189. A cationic lipid in Claim 1, which is DOTMA (1, 2-di-O- octadecenyl-3-trimethylammonium propane) or analog .

190. A cationic lipid in Claim 1, which is an esterified phosphatidylcholine .

191. A cationic lipid in Claim 1, which is 12 : 0 EPC ( 1 , 2-dilauroyl- sn-glycero-3-ethylphosphocholine) or analog.

192. A cationic lipid in Claim 1, which is 14 : 0 EPC ( 1, 2- dimyristoyl-sn-glycero-3-ethylphosphocholine) or analog.

193. A cationic lipid in Claim 1, which is 16 : 0 EPC ( 1, 2- dipalmitoyl-sn-glycero-3-ethylphosphocholine) or analog.

194. A cationic lipid in Claim 1, which is 18 : 0 EPC ( 1, 2- distearoyl-sn-glycero-3-ethylphosphocholine) or analog .

195. A cationic lipid in Claim 1, which is 14 : 1 EPC ( 1, 2- dimyristoleoyl-sn-glycero-3-ethylphosphocholine) or analog .

196. A cationic lipid in Claim 1, which is 18 : 1 EPC ( 1, 2-dioleoyl- sn-glycero-3-ethylphosphocholine) or analog.

197. A cationic lipid in Claim 1, which is 16 : 0-18 : 1 EPC ( 1- palmitoyl-2-oleoyl-sn-glycero-3-ethylphosphocholine) or analog .

198. A cationic lipid in Claim 1, which is a multivalent cationic lipid.

199. A cationic lipid in Claim 1, which is MVL5 (Nl- [ 2- ( ( IS ) -1- [ ( 3-aminopropyl) amino] -4- [di (3-amino- propyl) amino] butylcarboxamido) ethyl] -3, 4-di [oleyloxy] -benzamide) or analog.

200. A cationic lipid in Claim 1, which is a spermine-derived lipid.

201. A cationic lipid in Claim 1, which is DOSPA ( 2 , 3-dioleyloxy- N- [2- ( sperminecarboxamido) ethyl] -N, N-dimethyl-l-propanaminium) or analog .

202. A cationic lipid in Claim 1, which is GL67 (N4-Cholesteryl- Spermine) or analog.

203. A cationic lipid in Claim 1, which is DOGS (Dioctadecylamidoglycylspermine) or analog .204 . A cationic lipid in Claim 1, which is DPPES (Dipalmitoylphosphatidylethanolamidospermine) or analog .

205. A cationic lipid in Claim 1, which is RPR120535 or analog .

206. A cationic lipid in Claim 1, which is a cholesterol-based cationic lipid.207 . A cationic lipid in Claim 1, which is DC-Cholesterol • HC1 (3B- [N- (N ' , N ' -dimethylaminoethane) -carbamoyl ] cholesterol hydrochloride) or analog .208 . A cationic lipid in Claim 1, which is CLin-DMA (3- dimethylamino-2- ( cholest-5-en-3-beta-oxybutan-4-oxy) -1- (cis, cis- 9, 12-octadecadienoxy) propane) or analog.

209. A cationic or ionizable substance in any claim, which is comprised of a cationic polymer, block copolymer, copolymer, alternating copolymer, or random copolymer such as polyethyleneimine, poly (amidoamine) , or its derivative .210 . A polymer in Claim 1, which is a polyethylene glycol (PEG) - modified lipid.

211. A polymer in Claim 1, which is a PEG-modified lipid with a polyethylene glycol chain of up to 350 Da in length .

212. A polymer in Claim 1, which is 14 : 0 PEG350 PE (1, 2— dimy ri stoy 1-sn-gly cero-3 -pho sphoethano lamine-N- [methoxy (polyethylene glycol) -350 ] ) or analog.

213. A polymer in Claim 1, which is 16 : 0 PEG350 PE (1, 2— dipa Imitoy 1-sn-gly cero-3 -pho spho ethanolamine -N- [methoxy (polyethylene glycol) -350 ] ) or analog.

214. A polymer in Claim 1, which is 18 : 0 PEG350 PE ( 1 , 2-distearoyl- sn-glycero-3-phosphoethanolamine-N- [methoxy (polyethylene glycol) - 350 ] ) or analog.

215. A polymer in Claim 1, which is 18 : 1 PEG350 PE ( 1, 2-dioleoyl- sn-glycero-3-phosphoethanolamine-N- [methoxy (polyethylene glycol) - 350 ] ) or analog .

216. A polymer in Claim 1, which is a PEG-modified lipid with a polyethylene glycol chain of up to 550 Da in length .217 . A polymer in Claim 1, which is 14 : 0 PEG550 PE (DMPE-PEG550, 1 , 2 -dimy ri stoy 1-sn-gly cero-3 -pho sphoethano lamine-N- [methoxy (polyethylene glycol) -550 ] ) or analog.218 . A polymer in Claim 1, which is 16 : 0 PEG550 PE (1, 2- dipalmit oy 1-sn-gly cero-3 -pho spho ethanolamine -N- [methoxy (polyethylene glycol) -550 ] ) or analog.

219. A polymer in Claim 1, which is 18 : 0 PEG550 PE (DSPE-PEG550, 1, 2-distearoyl-sn-glycero-3-phosphoethanolamine-N- [methoxy (polyethylene glycol) -550 ] ) or analog.

220. A polymer in Claim 1, which is 18 : 1 PEG550 PE (DOPE-PEG550 , 1, 2-dioleoyl-sn-glycero-3-phosphoethanolamine-N- [methoxy (polyethylene glycol) -550 ] ) or analog.221 . A polymer in Claim 1 , which is a PEG-modified lipid with a polyethylene glycol chain of up to 750 Da in length .222 . A polymer in Claim 1 , which is 14 : 0 PEG750 PE ( 1 , 2- dimy ri stoy 1-sn-gly cero-3 -pho sphoethano lamine-N- [methoxy (polyethylene glycol ) -750 ] ) or analog .223 . A polymer in Claim 1 , which is 16 : 0 PEG750 PE ( 1 , 2- dipa Imitoy 1-sn-gly cero-3 -pho sphoethano famine -N- [methoxy (polyethylene glycol ) -750 ] ) or analog .224 . A polymer in Claim 1 , which is 18 : 0 PEG750 PE ( 1 , 2-distearoyl- sn-glycero-3-phosphoethanolamine-N- [methoxy (polyethylene glycol ) - 750 ] ) or analog .225 . A polymer in Claim 1 , which is 18 : 1 PEG750 PE ( 1 , 2-dioleoyl- sn-glycero-3-phosphoethanolamine-N- [methoxy (polyethylene glycol ) - 750 ] ) or analog .226 . A polymer in Claim 1 , which is a PEG-modified lipid with a polyethylene glycol chain of up to 1 kDa in length .227 . A polymer in Claim 1 , which is 14 : 0 PEG1000 PE (DMPE-PEG1000 , 1 . 2-dimyristoyl-sn-glycero-3-phosphoethanolamine-N- [methoxy (polyethylene glycol ) -1000 ] ) or analog .228 . A polymer in Claim 1 , which is 16 : 0 PEG1000 PE (DPPE-PEG1000 , 1 . 2 -dipalmitoyl-sn-glycero-3 -pho sphoethano lamine-N- [methoxy (polyethylene glycol ) -1000 ] ) or analog .229 . A polymer in Claim 1 , which is 18 : 0 PEG1000 PE (DSPE-PEG1000 , 1 . 2 -di st ear oy 1-sn-gly cero-3 -pho sphoethanolamine-N- [methoxy (polyethylene glycol) -1000 ] ) or analog .230 . A polymer in Claim 1 , which is 18 : 1 PEG1000 PE (DCPE-PEG1000 , 1 . 2 -diole oy 1-sn-gly cero-3 -pho sphoethano lamine-N- [methoxy (polyethylene glycol) -1000 ] ) or analog .231 . A polymer in Claim 1 , which is a PEG-modified lipid with a polyethylene glycol chain of up to 2 kDa in length .232 . A polymer in Claim 1 , which is 14 : 0 PEG2000 PE (DMPE-PEG2000 , 1 . 2-dimyristoyl-sn-glycero-3-phosphoethanolamine-N- [methoxy (polyethylene glycol) -2000 ] ) or analog .233 . A polymer in Claim 1 , which is 16 : 0 PEG2000 PE (DPPE-PEG2000 , 1 . 2 -dipalmitoyl- sn-glycer o-3 -phospho ethanolamine -N- [methoxy (polyethylene glycol ) -2000 ] ) or analog .234 . A polymer in Claim 1 , which is 18 : 0 PEG2000 PE (DSPE-PEG2000 , 1 . 2 -di st e aroyl- sn-glycer o- 3 -phosphoethanol amine-N- [methoxy (polyethylene glycol ) -2000 ] ) or analog .235 . A polymer in Claim 1 , which is DSPE-2arm-PEG2000 (N- [ 2 ' , 3 ' - Bis (methylpolyoxyethylene oxy) propane-1 ' -oxycarbonyl ] -1 , 2- distearoyl-sn-glycero-3-phosphoethanolamine) or analog .236 . A polymer in Claim 1 , which is DSPE-PEG2K-triGalNAc ( 1 , 2- di st e aroyl- sn-glycer o- 3 -phosphoethanol amine-N- [ carboxy (polyethylene glycol ) -2000 ] -N-tri (N-acetylgalactosamine ) ) or analog .237 . A polymer in Claim 1 , which is 18 : 1 PEG2000 PE (DCPE-PEG2000 , 1 . 2 -dioleoyl- sn-glycer o-3 -phosphoethanol amine-N- [methoxy (polyethylene glycol ) -2000 ] ) or analog .238 . A polymer in Claim 1 , which is a PEG-modified lipid with a polyethylene glycol chain of up to 3 kDa in length .239 . A polymer in Claim 1 , which is 14 : 0 PEG3000 PE ( 1 , 2- dimyristoyl-sn-glycero-3-phosphoethanolamine-N- [methoxy (polyethylene glycol ) -3000 ] ) or analog .240 . A polymer in Claim 1 , which is 16 : 0 PEG3000 PE ( 1 , 2 — dipalmi toy 1- sn-glycer o-3 -phospho ethanolamine -N- [methoxy (polyethylene glycol ) -3000 ] ) or analog .241 . A polymer in Claim 1 , which is 18 : 0 PEG3000 PE ( 1 , 2 — distearoyl-sn-glycero-3-phosphoethanolamine-N- [methoxy (polyethylene glycol ) -3000 ] ) or analog .242 . A polymer in Claim 1 , which is 18 : 1 PEG3000 PE ( 1 , 2-dioleoyl- sn-glycero-3-phosphoethanolamine-N- [methoxy (polyethylene glycol ) - 3000 ] ) or analog .243 . A polymer in Claim 1 , which is a PEG-modified lipid with a polyethylene glycol chain of up to 5 kDa in length .244 . A polymer in Claim 1 , which is 14 : 0 PEG5000 PE ( 1 , 2- dimy ri stoy 1-sn-gly cero-3 -pho sphoethano lamine-N- [methoxy (polyethylene glycol ) -5000 ] ) or analog .

245. A polymer in Claim 1, which is 16 : 0 PEG5000 PE ( 1, 2- dip Imitoyl-sn-gly cer o-3 -phospho ethanolamine -N- [methoxy (polyethylene glycol) -5000 ] ) or analog .

246. A polymer in Claim 1, which is 18 : 0 PEG5000 PE ( 1, 2- distearoyl-sn-glycero-3-phosphoethanolamine-N- [methoxy (polyethylene glycol) -5000 ] ) or analog .

247. A polymer in Claim 1, which is DSPE-2arm-PEG5000 (N- [2 ' , 3 ' - Bis (methylpolyoxyethylene oxy) propane-1 ' -oxycarbonyl] -1, 2- distearoyl-sn-glycero-3-phosphoethanolamine) or analog .

248. A polymer in Claim 1, which is 18 : 1 PEG5000 PE ( 1, 2-dioleoyl- sn-glycero-3-phosphoethanolamine-N- [methoxy (polyethylene glycol) - 5000 ] ) or analog .

249. A polymer in Claim 1, which is a PEG-modified glyceride .

250. A polymer in Claim 1, which is DMG-PEG2000 ( 1 , 2-dimyristoyl- rac-glycero-3-methoxypolyethylene glycol-2000) or analog.

251. A polymer in Claim 1, which is DSG-PEG2000 (a- [ 2 , 3-bis [ ( 1- oxooctadecyl) oxy] propyl] -w-methoxy-poly (oxy-1, 2-ethanediyl) ) or analog .252 . A polymer in Claim 1, which is DPG-PEG2000 ( 1 , 2-Dipalmitoyl- rac-glycero-3-methylpolyoxyethylene) or analog .

253. A polymer in Claim 1, which is DCG-PEG2000 ( 1 , 2-Dioleoyl-rac- glycerol, methoxypolyethylene Glycol) or analog.254 . A polymer in Claim 1, which is DMG-PEG5000 ( 1 , 2-Dimyristoyl- rac-glycero-3-methylpolyoxyethylene) or analog .

255. A polymer in Claim 1, which is DSG-PEG5000 ( 1, 2-Distearoyl- rac-glycero-3-methylpolyoxyethylene) or analog .

256. A polymer in Claim 1, which is DPG-PEG5000 ( 1 , 2-Dipalmitoyl- rac-glycerol, methoxypolyethylene Glycol) or analog.257 . A polymer in Claim 1, which is DCG-PEG5000 ( 1, 2-Dioleoyl-rac- glycerol, methoxypolyethylene Glycol) or analog .258 . A polymer in Claim 1, which is Sunbright GS-020MA (PEG2000 , 1, 2-Distearoyl-rac-glycero-3- (2 ' -maleimidoethyl) polyoxyethylene) or analog .

259. A polymer in Claim 1, which is Sunbright GS-020TS (PEG2000, 1, 2-Distearoyl-rac-glycero-3- ( succinimidyloxycarbonyl) polyoxyethylene) or analog.260 . A polymer in Claim 1, which is ALC-0159 (Methoxypoly ethylene glycol oxy (2000 ) -N, N-dit etradecyl acet amide) or analog .

261. A polymer in Claim 1, which is a PEG-modified sterol .262 . A polymer in Claim 1, which is Chol-PEG600 (Cholesterol- (polyethylene glycol-600 ) ) or analog .

263. A polymer in Claim 1, which is Chol-PEG2000 (Cholesterol- (polyethylene glycol-2000) ) or analog.264 . A polymer in Claim 1, which is a PEG-modified ceramide .

265. A polymer in Claim 1, which is C8 PEG750 ceramide (N-octanoyl- sphingosine-1- { succinyl [methoxy (polyethylene glycol) 750 ] } ) or analog .

266. A polymer in Claim 1, which is C16 PEG750 ceramide (N- palmitoyl-sphingosine-1- { succinyl [methoxy (polyethyleneglycol) 750 ] } ) or analog.267 . A polymer in Claim 1, which is C8 PEG2000 ceramide (N- octanoyl-sphingosine-1- { succinyl [methoxy (polyethyleneglycol) 2000 ] } ) or analog .268 . A polymer in Claim 1, which is C16 PEG2000 ceramide (N- palmitoyl-sphingosine-1- { succinyl [methoxy (polyethyleneglycol) 2000 ] } ) or analog .

269. A polymer in Claim 1, which is C8 PEG5000 ceramide (N- octanoyl-sphingosine-1- { succinyl [methoxy (polyethyleneglycol) 5000 ] } ) or analog .

270. A polymer in Claim 1, which is C16 PEG5000 ceramide (N- palmitoyl-sphingosine-1- { succinyl [methoxy (polyethyleneglycol) 5000 ] } ) or analog .

271. A polymer in Claim 1, which is a diacetylene lipid.

272. A polymer in Claim 1, which is 23 : 2 Diyne PE (1, 2-bis (10 , 12- tricosadiynoyl) -sn-glycero-3-phosphoethanolamine) or analog .

273. A polymer in Claim 1, which is 16 : 0-23 : 2 Diyne PE (PTPE, 1- palmitoyl-2- (10, 12-tricosadiynoyl) -sn-glycero-3- phosphoethanolamine) or analog .

274. A polymer in Claim 1, which is 23 : 2 Diyne PC ( 1 , 2-bis ( 10 , 12- tricosadiynoyl) -sn-glycero-3-phosphocholine) or analog.

275. A polymer in Claim 1, which is 16 : 0-23 : 2 Diyne PC (PTPC, 1- palmitoyl-2- (10, 12-tricosadiynoyl) -sn-glycero-3-phosphocholine) or analog.

276. A polymer in Claim 1, which is Tocofersolan (TPGS 1000, D-a- tocopheryl polyethylene glycol succinate) or analog.

277. An intracellular vesicle-interacting molecule in Claim 1, which is a cell-penetrating molecule .278 . An intracellular vesicle-interacting molecule in Claim 1, which is CPM2 or analog.

279. An intracellular vesicle-interacting molecule in Claim 1, which is a peptide .280 . An intracellular vesicle-interacting peptide in Claim 1, which is a membrane-disruptive peptide .

281. An intracellular vesicle-interacting peptide in Claim 1, which is aurein 1.2 (GLFDIIKKIAESF) or analog.282 . An intracellular vesicle-interacting peptide in Claim 1, which is a cell-penetrating peptide .

283. An intracellular vesicle-interacting peptide in Claim 1, which is an endosome-disrupting peptide .284 . An intracellular vesicle-interacting peptide in Claim 1, which is a truncated form of the N-terminus of adenovirus protein VI (LLLLAFSWGSLWSGIKNFGSTIKN) or analog .

285. An intracellular vesicle-interacting peptide in Claim 1, which is Tat (YGRKKRRQRRR) or analog.

286. An intracellular vesicle-interacting peptide in Claim 1, which is dTat-HA2 (D-Tat fused to HA2 ) or analog.287 . An intracellular vesicle-interacting peptide in Claim 1, which is transportan 10 (AGYLLGKINLKALAALAKKIL) or analog .288 . An intracellular vesicle-interacting peptide in Claim 1, which is octaarginine (R8 ) or analog.

289. An intracellular vesicle-interacting peptide in Claim 1, which is nonaarginine (R9) or analog .290 . An intracellular vesicle-interacting peptide in Claim 1, which is octalysine (L8) or analog .

291. An intracellular vesicle-interacting peptide in Claim 1, which is penetratin (RQIKIWFQNRRMKWKK) or analog .292 . An intracellular vesicle-interacting peptide in Claim 1, which is a histidine-swit ching cell-penetrating peptide .

293. An intracellular vesicle-interacting peptide in Claim 1, which is hsTAT (YGHKKHHQHHH) or analog.294 . An intracellular vesicle-interacting peptide in Claim 1, which is hsTPIO (FSGSGSGSGAGYLLGHINLHALAALAHHILGC) or analog.

295. An intracellular vesicle-interacting peptide in Claim 1, which is hsPen (FSGSGSGSGHQIHIWFQNHHMHWHHGC) or analog.

296. An intracellular vesicle-interacting peptide in Claim 1, which is hsTPT (CLIHHALAALAHLNIHLLYGASNLTWG) or analog.297 . An intracellular vesicle-interacting peptide in Claim 1, which is hsLMWP (GSVSHHHHHHGGHHHH) or analog.298 . An intracellular vesicle-interacting peptide in Claim 1, which is hsR12 (HHHHHHHHHHHH) or analog .

299. An intracellular vesicle-interacting peptide in Claim 1, which is hsMPG (GALFLGFLGAAGSTMGAWSQPHHHHHV) or analog.300 . An intracellular vesicle-interacting peptide in Claim 1, which is a fusogenic peptide .

301. An intracellular vesicle-interacting peptide in Claim 1, which is CM18 or analog .302 . An intracellular vesicle-interacting peptide in Claim 1, which is CM18-Tat (KWKLFKKIGAVLKVLTTG-YGRKKRRQRRR) or analog.

303. An intracellular vesicle-interacting peptide in Claim 1, which is diINF-7 or analog.304 . An intracellular vesicle-interacting peptide in Claim 1, which is H5WYG or analog.

305. An intracellular vesicle-interacting peptide in Claim 1, which is a pH-sensitive fusogenic peptide .

306. An intracellular vesicle-interacting peptide in Claim 1, which is HA2 ( GLFGAIAGFIENGWEGMIDGWYG) or analog .307 . An intracellular vesicle-interacting peptide in Claim 1, which is GALA ( WEAALAEALAEALAEHLAEALAEALEALAA) or analog.308 . An intracellular vesicle-interacting peptide in Claim 1, which is an amphiphilic peptide .

309. An intracellular vesicle-interacting peptide in Claim 1, which is 6His-CM18-PTD4 or analog .310 . An intracellular vesicle-interacting peptide in Claim 1, which is pHlip or analog.

311. An intracellular vesicle-interacting peptide in Claim 1, which is pHD118 or analog .312 . An intracellular vesicle-interacting peptide in Claim 1, which is a cyclic cell-penetrating peptide .

313. An intracellular vesicle-interacting peptide in Claim 1, which is cF4>R4 (cyclic phenylalanine-L-2-naphthylalanine- tetraarginine) or analog.314 . An intracellular vesicle-interacting peptide in Claim 1, which is CPP9 or analog .

315. An intracellular vesicle-interacting peptide in Claim 1, which is CPP12 or analog.

316. An intracellular vesicle-interacting peptide in Claim 1, which is palmitoyl-cyclic-dodeca-D-arginine (palmitoyl- GSSGCRRRRRRRRRRRRC-NH2) or analog .317 . An intracellular vesicle-interacting peptide in Claim 1, which is a miniature protein .318 . An intracellular vesicle-interacting peptide in Claim 1, which is miniature protein 5.3.

319. An intracellular vesicle-interacting peptide in Claim 1, which is miniature protein ZF5.3.320 . An intracellular vesicle-interacting peptide in Claim 1, which contains an endosomal escape domain .321 . An intracellular vesicle-interacting peptide in Claim 1, which contains the penetration accelerating sequence GKPILFF .

322. A cholesterol or cholesterol analog in Claim 1, which is cholesterol ( (3p) -cholest-5-en-3-ol) or analog .

323. A cholesterol or cholesterol analog in Claim 1, which is desmosterol (3B-hydroxy-5, 24-cholestadiene) or analog .324 . A cholesterol or cholesterol analog in Claim 1, which is lanosterol (lanosta-8 , 24-dien-3p-ol) or analog.

325. A cholesterol or cholesterol analog in Claim 1, which is lanosterol-95 ( 8, 24-lanostadien-3b-ol) or analog .

326. A cholesterol or cholesterol analog in Claim 1, which is 14- demethyl-lanosterol ( 4 , 4-dimethyl cholest-8 ( 9) , 24-dien-3B-ol) or analog .327 . A cholesterol or cholesterol analog in Claim 1, which is zymosterol (5a-cholesta-8 , 24-dien-3B-ol) or analog.328 . A cholesterol or cholesterol analog in Claim 1, which is lathosterol ( 5a-cholest-7-en-3B-ol) or analog .

329. A cholesterol or cholesterol analog in Claim 1, which is 24- methylene cholesterol (5, 24 (28 ) -cholestadien-24-methylen-3t>-ol) or analog .330 . A cholesterol or cholesterol analog in Claim 1, which is a dehydrocholesterol .

331. A cholesterol or cholesterol analog in Claim 1, which is 7- dehydrocholesterol (A5, 7-cholesterol) or analog .332 . A cholesterol or cholesterol analog in Claim 1, which is 8 ( 9) - dehydrocholesterol (cholesta-5, 8 ( 9) -dien-3B-ol) or analog .

333. A cholesterol or cholesterol analog in Claim 1, which is 8 (14) -dehydrocholesterol (cholesta-5, 8 ( 14 ) -dien-3B-ol) or analog.334 . A cholesterol or cholesterol analog in Claim 1, which is FF- MAS ( 14-demethyl-14-dehydrolanosterol) or analog.

335. A cholesterol or cholesterol analog in Claim 1, which is a saturated sterol .

336. A cholesterol or cholesterol analog in Claim 1, which is dihydrolanosterol (24 , 25-dihydrolanosterol) or analog .337 . A cholesterol or cholesterol analog in Claim 1, which is zymostenol (5 e -Cholest-8-en-3-beta-ol) or analog.

338. A cholesterol or cholesterol analog in Claim 1, which is a cholesterol ester .

339. A cholesterol or cholesterol analog in Claim 1, which is cholesteryl nervonate (cholest-5-en-3-ol, 3p- [ (15Z) -15- tetracosenoate ] ) or analog .340 . A cholesterol or cholesterol analog in Claim 1, which is cholesteryl oleate (cholest-5-en-3p-ol, 3- [ ( 9Z ) -9-octadecenoate ] ) or analog .

341. A cholesterol or cholesterol analog in Claim 1, which is cholesteryl palmitate ( (3p) -cholest-5-en-3-ol, 3-hexadecanoate) or analog .342 . A cholesterol or cholesterol analog in Claim 1, which is 17 : 0 cholesteryl ester (cholest-5-en-3f>-yl heptadecanoate) or analog .

343. A cholesterol or cholesterol analog in Claim 1, which is cholesteryl stearate ( (3p) -cholest-5-en-3-ol, 3-octadecanoate) or analog .344 . A cholesterol or cholesterol analog in Claim 1, which is cholesteryl lignocerate (cholest-5-en- (3p) -ol, 3-tetracosanoate) or analog .

345. A cholesterol or cholesterol analog in Claim 1, which is cholesteryl homo-y_linolenate (cholest-5-en-3p-ol, 3- ( 8Z, 11Z, 14Z) - 8 , 11, 14-eicosatrienoate) or analog .

346. A cholesterol or cholesterol analog in Claim 1, which is an oxysterol .347 . A cholesterol or cholesterol analog in Claim 1, which is ip- hydroxy cholesterol (cholest-5-ene-3p, 4 p-diol) or analog.348 . A cholesterol or cholesterol analog in Claim 1, which is 25- hydroxy cholesterol (cholest-5-ene-3p, 25-diol) or analog.

349. A cholesterol or cholesterol analog in Claim 1, which is a phytosterol (derived from plants) .350 . A cholesterol or cholesterol analog in Claim 1, which is stigmasterol ( stigmasta-5 , 22E-dien-3p-ol) or analog .

351. A cholesterol or cholesterol analog in Claim 1, which is 24- methyl cholesterol ( (3p, 24 ^) -ergost-5-en-3-ol) or analog.

352. A cholesterol or cholesterol analog in Claim 1, which is p- Sitosterol ( stigmast-5-en-3p-ol) or analog .

353. A cholesterol or cholesterol analog in Claim 1, which is brassicasterol (ergosta-5, 22E-dien-3p-ol) or analog.354 . A cholesterol or cholesterol analog in Claim 1, which is fucosterol ( (3p, 24E) -stigmasta-5, 24 (28 ) -dien-3-ol) or analog .

355. A cholesterol or cholesterol analog in Claim 1, which is a- spinasterol ( (3p, 5a, 22E) -stigmasta-7 , 22-dien-3-ol) or analog .

356. A cholesterol or cholesterol analog in Claim 1, which is campesterol ( campest-5-en-3f>-ol) or analog .357 . A cholesterol or cholesterol analog in Claim 1, which is a saturated phytosterol .358 . A cholesterol or cholesterol analog in Claim 1, which is sitostanol ( ( 5a) -stigmastan-3p-ol) or analog.

359. A cholesterol or cholesterol analog in Claim 1, which is campestanol ( (3p, 5a, 24R) -ergostan-3-ol) or analog .360 . A cholesterol substitute in Claim 1, which is a sterol or sterol derivative .

361. A cholesterol substitute in Claim 1, which is a chemical or biological substance that can modify lipid membrane fluidity or rigidity over a range of physiological temperatures .362 . A cholesterol substitute in Claim 1, which contains at least one polar group such as a hydroxyl, at least one segment comprised of at least 3 aliphatic rings, and at least one hydrophobic group comprised of at least 3 and at most 16 carbon atoms .

363. A targeting element in Claim 1, which is an antibody.364 . A targeting element in Claim 1, which is an antibody derivative such as a nanobody, antibody fragment, bispecific, antibody variable region or a combination of one or more antibody derivatives .

365. A targeting element in Claim 1, which is anti-c-Kit (anti- CD117 ) .

366. A targeting element in Claim 1, which is anti-glucocorticoid receptor .

367. A targeting element in Claim 1, which is anti-transforming growth factor beta receptor 2 (TGFBR2) .368 . A targeting element in Claim 1, which is anti-transforming growth factor beta receptor 3 (TGFBR3) .

369. A targeting element in Claim 1, which is anti-aquaporin 7 .370 . A targeting element in Claim 1, which is anti-aquaporin 7B .

371. A targeting element in Claim 1, which is anti-activin A receptor type 1C .372 . A targeting element in Claim 1, which is anti-frizzled class 4 receptor .

373. A targeting element in Claim 1, which is anti-perilipin 1.374 . A targeting element in Claim 1, which is anti-perilipin 4 .

375. A targeting element in Claim 1, which is anti-SLC7A10 .

376. A targeting element in Claim 1, which is anti-platelet- derived growth factor receptor alpha .377 . A targeting element in Claim 1, which is anti-platelet- derived growth factor receptor beta .378 . A targeting element in Claim 1, which is a nanobody .

379. A targeting element in Claim 1, which is an antigen-binding fragment (Fab) .380 . A targeting element in Claim 1, which is a bispecific or multi-specific antibody or antibody fragment .

381. A targeting element in Claim 1, which is comprised at least in part of an antibody variable region .382 . A targeting element in Claim 1, which is a protein .

383. A targeting element in Claim 1, which is angiotensin II . 384 . A targeting element in Claim 1, which is VEGF .

385. A targeting element in Claim 1, which is wheat germ agglutinin (WGA) .

386. A targeting element in Claim 1, which is Ulex europaeus agglutinin I (UEA-1) .387 . A targeting element in Claim 1, which is a peptide .388 . A targeting element in Claim 1, which is miniAp-4 .

389. A targeting element in Claim 1, which is PEP3 .

390. A targeting element in Claim 1, which is a peptide with the sequence CKGGRAKDC .

391. A targeting element in Claim 1, which is a peptide with the sequence GGGGYDRVTIHPF .

392. A targeting element in Claim 1, which is a peptide with the sequence D (KLAKLAK) 2 .

393. A targeting element in Claim 1, which is a peptide with the sequence MRYMILGLLALAAVCSA .

394. A targeting element in Claim 1, which is a receptor ligand.

395. A targeting element in Claim 1, which is LJ-4378 (agonist of A2A adenosine receptor / antagonist of A3adenosine receptor) .

396. A targeting element in Claim 1, which is resistin (ligand of decorin lacking the glycanation site) .

397. A targeting element in Claim 1, which is norrin ( specific ligand of frizzled class 4 receptor) .

398. A targeting element in Claim 1, which is a receptor binding domain .

399. A targeting element in Claim 1, which is a lipid.

400. A targeting element in Claim 1, which is PAHSA (palmitic- acid-9-hydroxy-stearic-acid) or analog .

401. A targeting element in Claim 1, which is a polysaccharide .402 . A targeting element in Claim 1, which is glycogen .

403. A targeting element in Claim 1, which is glucan .404 . A targeting element in Claim 1, which is dextran .

405. A targeting element in Claim 1, which is chitosan .

406. A targeting element in Claim 1, which is alginate .407 . A targeting element in Claim 1, which is hyaluronic acid. 408 . A targeting element in Claim 1, which is a nucleic acid.

409. A targeting element in Claim 1, which is an aptamer .410 . A targeting element in Claim 1, which is an aptamer against VEGFR.

411. A nucleic acid in Claim 1, which is a double-stranded deoxyribonucleic acid (DNA) .412 . A nucleic acid in Claim 1, which is a single-stranded DNA.

413. A nucleic acid in Claim 1, which is a linear DNA.

414. A nucleic acid in Claim 1, which is a circular DNA.

415. A nucleic acid in Claim 1, which is a DNA 0. 5 to 50 kb in length .

416. A nucleic acid in Claim 1, which is a DNA 1. 5 to 7.0 kb in length .417 . A genetic cargo in Claim 1, which encodes at least a promoter and / or enhancer .418 . A genetic cargo in Claim 1, which encodes at least the cytomegalovirus (CMV) immediate early promoter .

419. A genetic cargo in Claim 1, which encodes at least the eukaryotic translation elongation factor 1 al (EF1A) promoter .

420. A genetic cargo in Claim 1, which encodes at least the eukaryotic translation elongation factor 1 al short form (EFS) promoter .

421. A genetic cargo in Claim 1, which encodes at least the CAG promoter .

422. A genetic cargo in Claim 1, which encodes at least the CBh promoter .

423. A genetic cargo in Claim 1, which encodes at least the CBA promoter .

424. A genetic cargo in Claim 1, which encodes at least the simian virus 40 (SV40 ) enhancer / early promoter .

425. A genetic cargo in Claim 1, which encodes at least the phosphoglycerate kinase 1 (BGK) promoter .

426. A genetic cargo in Claim 1, which encodes at least an adiposespecific promoter .

427. A genetic cargo in Claim 1, which encodes at least the adipocyte B2 (aP2) promoter .

428. A genetic cargo in Claim 1, which encodes at least the adiponectin promoter .

429. A genetic cargo in Claim 1, which encodes at least the peroxisome prolif erator-activated receptor gamma (PEAR gamma) promoter .

430. A genetic cargo in Claim 1, which encodes at least the fatty acid binding protein 4 (FABP4) promoter .

431. A genetic cargo in Claim 1, which encodes at least the laeverin (LVRN) promoter .

432. A genetic cargo in Claim 1, which encodes at least the perilipin 1 (PLIN1) promoter .

433. A genetic cargo in Claim 1, which encodes at least the perilipin 4 (PLIN4 ) promoter .

434. A genetic cargo in Claim 1, which encodes at least a therapeutic transgene .

435. A genetic cargo in Claim 1, which encodes at least insulin .

436. A genetic cargo in Claim 1, which encodes at least gastric inhibitory peptide (GIP) receptor agonist or analog.

437. A genetic cargo in Claim 1, which encodes at least glucagon- like peptide 1 (GLP1) receptor agonist or analog .

438. A genetic cargo in Claim 1, which encodes at least a GIP / GLP1 receptor co-agonist or analog.

439. A genetic cargo in Claim 1, which encodes at least insulinlike growth factor 1 ( IGF1) .

440. A genetic cargo in Claim 1, which encodes at least insulinlike growth factor 2 ( IGF2 ) .

441. A genetic cargo in Claim 1, which encodes at least beta- glucocerebrosidase .

442. A genetic cargo in Claim 1, which encodes at least an alphaglucosidase .

443. A genetic cargo in Claim 1, which encodes at least fibroblast growth factor 18 (FGF18) .

444. A genetic cargo in Claim 1, which encodes at least vascular endothelial growth factor (VEGFA) .

445. A genetic cargo in Claim 1, which encodes at least factor VIII .

446. A genetic cargo in Claim 1, which encodes at least factor IX .

447. A genetic cargo in Claim 1, which encodes at least liver- expressed antimicrobial peptide 2 (LEAP2 , MTPFWRGVSLRP IGASCRDDSECITRLCRKRRCSLSVAQE ) .

448. A genetic cargo in Claim 1, which encodes at least an siRNA.

449. A genetic cargo in Claim 1, which encodes at least an siRNA against X-linked inhibitor of apoptosis-associated factor 1 (XAF1) .

450. A genetic cargo in Claim 1, which encodes at least an shRNA.

451. A genetic cargo in Claim 1, which encodes at least an antibody .

452. A genetic cargo in Claim 1, which encodes at least an antibody fragment .

453. A genetic cargo in Claim 1, which encodes at least a nanobody .

454. A genetic cargo in Claim 1, which encodes at least an antitumor necrosis factor alpha (TNF-alpha) nanobody .

455. A genetic cargo in Claim 1, which encodes at least a post- transcriptional regulatory element .

456. A genetic cargo in Claim 1, which encodes at least the hepatitis B post-transcriptional regulatory element (HPRE) .

457. A genetic cargo in Claim 1, which encodes at least the Woodchuck Hepatitis Virus post-transcriptional regulatory element (WPRE) .

458. A genetic cargo in Claim 1, which encodes at least the optimized Woodchuck Hepatitis Virus post-transcriptional regulatory element (oPRE) .

459. A genetic cargo in Claim 1, which encodes at least polyadenylation sequence .

460. A genetic cargo in Claim 1, which encodes at least the bovine growth hormone polyadenylation sequence (bGH pA) .

461. A genetic cargo in Claim 1, which encodes at least the SV40 polyadenylation sequence (SV40 pA) .

462. A genetic cargo in Claim 1, which encodes at least a downtitration element .

463. A genetic cargo in Claim 1, which encodes at least a suicide gene .

464. A genetic cargo in Claim 1, which encodes iCasp8 (inducible caspase 8) .

465. A genetic cargo in Claim 1, which encodes iCasp9 (inducible caspase 9) .

466. A nanoparticle formulation in Claim 1, with an average diameter of 25 - 250 nm.

467. A nanoparticle formulation in Claim 1, with an average diameter of 35 - 150 nm.

468. A nanoparticle formulation in Claim 1, with a zeta potential in the range of -0 .5 to -15. 0 mV.

469. A nanoparticle formulation in Claim 1, with a zeta potential in the range of -6.0 to -10. 0 mV.

470. A nanoparticle formulation in Claim 1, with a polydispersity index of 0 .001 to 2. 50 .

471. A nanoparticle formulation in Claim 1, with a polydispersity index of 0 .01 to 0 .3.

472. A nanoparticle formulation in Claim 1, with an N to P ratio of 1.5 to 20 .

473. A nanoparticle formulation in Claim 1, with an N to P ratio of 3.5 to 9. 5.

474. A nanoparticle formulation in Claim 1, with ionizable lipid ( s) comprising a molar ratio of about 15-65% .

475. A nanoparticle formulation in Claim 1, with structural element ( s) comprising molar ratio of about 0-35% of the total lipid present in the formulation .

476. A nanoparticle formulation in Claim 1, with polymer ( s) comprising a molar ratio of about 0-20% of the total lipid present in the formulation .

477. A nanoparticle formulation in Claim 1, with sterol (s) , cholesterol ( s) , or cholesterol-like molecule ( s) comprising a molar ratio of about 0-55% of the total lipid present in the formulation .

478. A nanoparticle formulation in Claim 1 to treat diabetes .

479. A nanoparticle formulation in Claim 1 to treat growth restriction .

480. A nanoparticle formulation in Claim 1 to treat rheumatoid arthritis .

481. A nanoparticle formulation in Claim 1 to treat Gaucher' s disease .

482. A nanoparticle formulation in Claim 1 to treat progeria .483 . A nanoparticle formulation in Claim 1 to treat Pompe disease .484 . A nanoparticle formulation in Claim 1 to treat hemophilia .

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