Lipids and lipid nanoparticles

Abstract

The present invention provides for lipids that may be formulated in a delivery vehicle to facilitate the encapsulation of a wide range of payloads including therapeutic and diagnostic agents, such as, without limitation, nucleic acids (e.g., RNA or DNA), proteins, peptides, and small molecule active pharmaceutical ingredients (APIs).

Description

Lipids and Lipid NanoparticlesCross-Reference to Related ApplicationsThis application claims the benefit of U.S. Provisional Application Serial No. 63 / 536,997, filed September 7, 2023, and U.S. Provisional Application Serial No. 63 / 664,820, filed June 27, 2024, the disclosure of which is hereby incorporated by reference in its entirety, including all figures, tables and amino acid or nucleic acid sequences.Background of the inventionLipid nanoparticles (LNPs) are a type of nanoscale delivery system composed of lipids that have gained significant attention and validation in the field of medicine. These particles have shown great potential for the delivery of various therapeutic agents, including nucleic acids such as DNA, RNA, and siRNA. LNPs offer several advantages over other delivery systems, including intracellular delivery of sensitive nucleic acids like mRNA, biocompatibility, biodegradability, and the ability to encapsulate hydrophobic and hydrophilic molecules.The development of LNPs as a drug delivery system can be traced back to the early 1990s. Researchers recognized the potential of lipids to form stable nanoparticles and protect encapsulated molecules. The initial focus of LNP research was on gene therapy, where the delivery of nucleic acids posed significant challenges due to their inherent instability and the need for efficient intracellular delivery. Over the years, advancements in lipid chemistry, formulation techniques, and manufacturing processes have contributed to the refinement of LNP-based delivery systems.LNPs have shown tremendous promise in the field of medicine, particularly in the delivery of nucleic acid-based therapeutics. This includes the delivery of small interfering RNA (siRNA) for gene silencing, messenger RNA (mRNA) for protein synthesis, and gene editing tools such as CRISPR-Cas9. LNPs protect the encapsulated nucleic acids from degradation, enhance their cellular uptake, and facilitate their release at the target site, thereby improving therapeutic efficacy.One of the significant breakthroughs in LNP-based medicine came with the development of COVID-19 mRNA vaccines. The mRNA vaccines from Pfizer-BioNTech and Moderna, which use LNPs as delivery vehicles, demonstrated remarkable effectiveness in combating COVID- 19. These vaccines not only showcased the potential of LNPs but also paved the way for the rapid development and deployment of mRNA-based vaccines for other diseases.Despite the progress made, there are several hurdles and problems that remain with LNP- based delivery systems. One of the main challenges is achieving efficient and targeted delivery to specific tissues or cells. LNPs face barriers such as uptake by thereticuloendothelial system (RES) and clearance by the liver and kidneys, which can limit their circulation time and reduce their therapeutic efficacy.Another challenge is the potential for toxicity and immune responses. LNPs may induce an immune response due to their foreign nature, leading to adverse reactions or decreased therapeutic outcomes.Furthermore, the scalability and manufacturing of LNPs pose significant challenges. Maintaining batch-to-batch consistency, controlling particle size, and optimizing production processes are crucial for the successful translation of LNPs from the laboratory to large-scale manufacturing.There exists a need to develop new lipid compounds, e.g., cationic lipids, ionizable lipids (generally amines, tertiary substituted), polymer-conjugated lipids, and structural lipids, that afford efficient delivery of the therapeutic agents, sufficient activity of the therapeutic agents (e.g., expression of mRNA after delivery), optimal pharmacokinetics, and / or other suitable physiological, biological, and / or therapeutic properties.Brief Summary of the InventionThe present invention provides for lipids that may be formulated in a delivery vehicle to facilitate the encapsulation of a wide range of payloads including therapeutic and diagnostic agents, such as, without limitation, nucleic acids (e.g., RNA or DNA), proteins, peptides, and small molecule active pharmaceutical ingredients (APIs).The lipid compounds of the present invention can be used in combination with other lipid components, such as neutral lipids, sterols and polymer conjugated lipids, to form lipid nanoparticles for delivery of payloads both in vitro and in vivo, for therapeutic or prophylactic purposes, including vaccination. Thus, the present invention further provides for lipid nanoparticles (LNPs) comprising said lipids as well as methods of administering LNPs to a subject, e.g., delivering an mRNA and achieving prolonged expression of a desired polypeptide in the animal or human subject.In further embodiments, the invention provides for a method of delivering and / or producing a polypeptide of interest in a cell.In further embodiments, the invention provides for a method of treating a disease, disorder, or condition in a subject, comprising the step of administering the foregoing lipid nanoparticle and / or lipid nanoparticle composition, to a subject in need of such treatment. The lipid particle and / or lipid particle composition may be also delivered to a subject as a component of a vaccine or diagnostic composition.Brief Description of the DrawingsFIGs. 1A-1B. Size assessed by Dynamic Light Scattering (DLS) of LNPs with and without fifth component. FIG. 1A) MC3 LNPs with 45mol% MC3 without and with AAP modifiers (50% denotes a control without AAP and having 50mol % MC3) with compositions according toTable 1. FIG. IB) Size assessed by DLS of LNPs with compositions according to Table 1. MC3 is the chemical named (6Z,9Z,28Z,31Z)-heptatriaconta-6,9,28,31-tetraen-19-yl 4- (dimethylamino)butanoate.FIGs. 2A-2B. Polydispersity of LNPs with and without fifth component. FIG. 2A) MC3 LNPs with 45mol% MC3 without and with AAP modifiers (50% denotes a control without AAP and containing 50mol % MC3). FIG. 2B) Polydispersity of LNPs with compositions and structures of ionizable lipids per Table 1.FIGs. 3A-3B. Size and polydispersity of LNPs with and without fifth component, on day 14 after formulation and storage in refrigerator. FIG. 3A) MC3 LNP sizes with 45mol% MC3 without and with AAP modifiers (50% denotes a control without AAP and containing 50mol % MC3) according to Table 1. FIG. 3B) Polydispersity of LNPs with compositions according to Table 1.FIGs. 4A-4B. In vitro expression of Firefly Luciferase in HEK293 cells, measured by luminescence, expressed as relative light units (RLU). MC3 LNPs with 45mol% MC3 with AAP and without modifiers (FIG. 4A). Novel ionizable lipid-based LNPs with same mRNA (FIG. 4B) show proportional increase in luminescence with increased dose. Compositions are according to Table 1.FIGs. 5A-5B. In vitro expression of Firefly Luciferase in Huh7 cells, measured by luminescence, expressed as relative light units (RLU). MC3 LNPs with 45mol% MC3 with AAP and without modifiers (FIG. 5A). Novel ionizable lipid-based LNPs with same mRNA (FIG. 5B) show proportional increase in luminescence with increased dose. Compositions are according to Table 1.FIGs. 6A-6B. Viability of cells in the in vitro expression of Firefly Luciferase in HEK293T cells, expressed as relative fluorescence units (RFU) using TiterGlo kit. MC3 LNPs with 45mol% MC3 with AAP and without modifiers (FIG. 6A). Novel ionizable lipid-based LNPs with same mRNA (FIG. 6B) show proportional increase in luminescence with increased dose. Compositions are according to Table 1.FIGs. 7A-7B. Viability of cells in the in vitro expression of Firefly Luciferase in Huh7 cells, expressed as relative fluorescence units (RFU) using TiterGlo kit. IVIC3 LNPs with 45mol% IVIC3 with AAP and without modifiers (FIG. 7A). Novel ionizable lipid-based LNPs with same mRNA (FIG. 7B) show proportional increase in luminescence with increased dose. Compositions are according to Table 1.FIG. 8. Fluorescence emission of LNP formulations in Table 2. Irradiation of the pure Compound (89) in ethanol at 480 nm is associated with emission band at around 560 nm. Irradiation of the LNP formulations at 480 nm is associated with negligible emission at around 560 nm, showing fluorescence quenching.FIG. 9. Size and polydispersity by Dynamic Light Scattering (DLS) of LNPs with and without fifth component Compound (89) and astaxanthin. Left side: Z-average size of LNPs. Right side: Polydispersity of LNPs with compositions and structures of ionizable lipids per Table 2. Composition labels are according to Table 1.FIG. 10. Expression of Luciferase from modified mRNA by in vitro cell transfection with LNPs with varying amounts of Compound (89) additive in HEK293T cells (left) and Huh7 cells (right). Compositions are according to Table 1.FIG. 11. Viability of cells tested with mRNA LNPs in vitro cell transfection, with formulations in Table 2 having varying amounts of Compound (89) additive in HEK293T cells (left) and Huh7 cells (right). Compositions are according to Table 1.FIGs. 12A-12B. Characterization of LNPs containing MC3 and ionizable lipids Compound 51 or Compound 53, with additional components as per Table 2. FIG. 12A: Average Z diameter; FIG. 12B: Polydispersity (PDI).FIGs. 13A-13B. Expression of Luciferase from modified mRNA with LNPs containing Compound 51 or Compound 53 by in vitro cell transfection of HEK293T cells (FIG. 13A) and Huh7 cells (FIG. 13B). LP-01 is the chemical named 3-((4,4-bis(octyloxy)butanoyl)oxy)-2-((((3- (diethylamino)propoxy)carbonyl)oxy)methyl)propyl (9Z,12Z)-octadeca-9,12-dienoate.FIG. 14. Expression of Luciferase from modified mRNA with LNPs (see Table 3) containing modified mRNA encoding Luciferase and ionizable lipid Compound 30 or Compound 52b y in vivo intravenous dosing in Balb / C mice.Detailed Description of the InventionDefinitions"Administration", "administering" and variants thereof refers to introducing a composition or agent of the present invention (e.g., an LNP comprising a nucleic acid payload) into a subject, organ, tissue or cells for therapeutic, pharmacokinetic, diagnostic purposes, or research purposes. "Administration" includes in vivo, in vitro, ex vivo and in utero administration. The introduction of a composition or agent into a subject is by any route of administration that is suitable for the specific composition or agent. Routes of administration include oral, pulmonary, intranasal, parenteral (intravenous, intramuscular, intraperitoneal, or subcutaneous), rectally, intravesical, intranodal, intralymphatical, intratumoral and topical administration.The term "cationic lipid" refers to any lipid that is or can be positively charged. In some embodiments, the cationic lipid is an ionizable lipid, i.e., an ionizable cationic lipid, which is predominantly protonated and positively charged at a pH that exceeds about 2 units of the pKa value, known as the negative log(10) value of the acid dissociation constant (Ka) of the protonated, positively charged form of the lipid. Cationic and cationic ionizable lipids are highly water-insoluble, even when charged, due to the lipophilic nature of the overallmolecule. Therefore, the thermodynamic pKa of an ionizable lipid is generally not measured but is estimated or calculated, and the measurement of pKa is instead done at the particle level by a TNS method (Heyes J et al., J Control Release 2005, pp. 276-287). A pKa measured in this way, on LNPs containing among other lipids the ionizable lipid, is termed "apparent pKa", "pKa(app)" or "LNP pKa" and is generally denoted as pKa'. The pKa' is about 2 to 4 units lower than the pKa, based on estimation methodology proposed for correlating the pKa and pKa' (e.g. Carrasco et al., Comm Biol, 2021, vol 4:956). For the purposes of relating functional aspects of ionizable lipids and their LNPs we use pKa' in the following sections.One function of ionizable lipids is to facilitate endosomal release, by changing LNP structure via interaction with the endosomal membrane of LNPs within endosomes. This occurs about pH 5.5 and below. In some embodiments, ionizable cationic lipid in an LNP is predominantly neutral at physiological pH (pH 7.4) and is ionized and positively charged in the same LNP at a pH that is below physiological pH (e.g., between pH 4-5, preferably about pH 4.5). In some embodiments, the ionizable cationic lipid is about neutral at physiological pH (7.4 pH) and becomes protonated when introduced to an environment where the pH is about 4.5 or the pH inside of endosomes. It will be understood by one of ordinary skill in the art that the addition or removal of protons as a function of pH is a fast equilibrium process, and that the reference to a charged or a neutral lipid refers to the nature of the predominant species and does not require that all of the lipid be present in the charged or neutral form, whether in isolation or in an LNP. Generally, ionizable cationic lipids have a pKa' in the range of about 4 to about 7 as measured in the particle context (TNS assay). In some embodiments, ionizable lipid may include "cleavable lipid" or "SS-cleavable lipid". Corresponding quaternary lipids of all ionizable cationic lipids described herein (i.e., where a nitrogen atom is protonated and has four substituents) are contemplated within the scope of this disclosure. In some embodiments, an LNP comprising an ionizable lipid has a an pKa' (TNS) between 4-5, 5-6 between 4-5, 5-6, or 6-7. A "non-cationic lipid" is an anionic or neutral lipid.The term "anionic lipid" refers to any lipid that is negatively charged at pH 7.4 (physiological pH). These lipids include, but are not limited to, phosphatidylglycerols, cardiolipins, diacylphosphatidylserines, diacylphosphatidic acids, N-dodecanoyl phosphatidylethanolamines, N-succinyl phosphatidylethanolamines, N- glutarylphosphatidylethanolamines, lysylphosphatidylglycerols, palmitoyloleyolphosphatidylglycerol (POPG), and other anionic modifying groups joined to neutral lipids.The term "hydrophobic lipid" refers to compounds having a polar group(s) that include, but are not limited to, long-chain saturated and unsaturated aliphatic hydrocarbon groups and such groups optionally substituted by one or more aromatic, cycloaliphatic, or heterocyclic group(s). Suitable examples include, but are not limited to, diacylglycerol, dialkylglycerol, N,N-dialkylamino, l,2-diacyloxy-3-aminopropane, and l,2-dialkyl-3-aminopropane.The term "hydroxyalkyl" means a linear monovalent hydrocarbon radical or a branched monovalent hydrocarbon radical, substituted with one or two hydroxy groups, provided that if two hydroxy groups are present, they are not on the same carbon atom. Representative examples include, but are not limited to, hydroxymethyl, 2-hydroxyethyl, 2-hydroxypropyl, 3-hydroxypropyl, l-(hydroxymethyl)-2- methylpropyl, 2-hydroxybutyl, 3-hydroxybutyl, 4- hydroxybutyl, 2,3-dihydroxypropyl, 1- (hydroxymethyl)-2-hydroxyethyl, 2,3-dihydroxybutyl, 3,4-dihydroxybutyl and 2- (hydroxymethyl)-3-hydroxypropyl, preferably 2- hydroxyethyl, 2,3-dihydroxypropyl, and 1- (hydroxymethyl)-2-hydroxyethyl. A C1-C6 hydroxyalkyl means a linear monovalent hydrocarbon radical of one to six carbon atoms or a branched monovalent hydrocarbon radical of three to six carbons substituted with either one hydroxy group or two hydroxy groups on different carbon atoms. Where the alkyl is substituted with an alkene, the group is a "hydroxyalkenyl" group.The term "linked" encompasses chemical conjugation, adsorption (physisorption and / or chemisorption). The types of bonds encompassed by the term "linked" are covalent interactions and noncovalent interactions (e.g., hydrogen bonds, ionic bonds, van der Waal bonds, and hydrophobic bonds).The term "neutral lipid" refers to a lipid that exist either in an uncharged or neutral zwitterionic form in a pH range comprising pH 4 - 7.4. At physiological pH, such lipids include, for example, diacylphosphatidylcholine, diacylphosphatidylethanolamine, ceramide, sphingomyelin, cephalin, cholesterol, cerebrosides, and diacylglycerols.The term "non-fusogenic cationic lipid" is meant a cationic lipid that can condense and / or encapsulate the nucleic acid cargo, such as mRNA, but does not have, or negligible, fusogenic activity with a cell plasma membrane.The term "cleavable lipid" or "SS-cleavable lipid" refers to a lipid comprising a disulfide bond cleavable unit. Cleavable lipids may include cleavable disulfide bond ("SS") containing lipid- like materials that comprise a pH-sensitive tertiary amine and self-degradable phenyl ester. For example, a SS-cleavable lipid can be an SS-OP lipid (COATSOME® SS-OP), an SS-M lipid (COATSOME® SS-M), an SS-E lipid (COATSOME® SS-E), an SS-EC lipid (COATSOME® SS-EC), an SS-LC lipid (COATSOME® SS-LC), an SS-OC lipid (COATSOME® SS-OC), and an SS-PalmE lipid (see, for example, Formulae l-IV), or a lipid described in Togashi R, et al. J Control Release 2018 Jun 10;279:262-270, US Patent 9,708,628, or US Patent 10,385,030.The term "non-cationic lipid" refers to a neutral lipid or anionic lipid.The term "nucleic acid," refers to a polymer containing at least two nucleotides (i.e., deoxyribonucleotides or ribonucleotides) in either single- or double-stranded form and includes DNA, RNA, DNA-RNA hybrids, as well as analogs and modified forms thereof. DNA may be in the form of, e.g., antisense molecules, plasmid DNA, DNA-DNA duplexes, precondensed DNA, PCR products, vectors (PI, PAC, BAC, YAC, artificial chromosomes), expression cassettes, chimeric sequences, chromosomal DNA, or derivatives andcombinations of these groups. DNA may be in the form of minicircle, plasmid, bacmid, minigene, ministring DNA (linear covalently closed DNA vector), closed-ended linear duplex DNA (CELiD or ceDNA), doggybone™ DNA, dumbbell shaped DNA, minimalistic immunological-defined gene expression (MIDGE)-vector, viral vector or nonviral vectors. RNA may be in the form of small interfering RNA (siRNA), dicer-substrate dsRNA, small hairpin RNA (shRNA), asymmetrical interfering RNA (aiRNA), microRNA (miRNA), mRNA, rRNA, tRNA, gRNA, viral RNA (vRNA), and combinations thereof. Nucleic acids include nucleic acids containing known nucleotide analogs or modified backbone residues or linkages, which are synthetic, naturally occurring, and non-naturally occurring, and which have similar binding properties as the reference nucleic acid. Examples of such analogs and / or modified residues include, without limitation, phosphorothioates, phosphorodiamidate morpholino oligomer (morpholino), phosphoramidates, methyl phosphonates, chiral-methyl phosphonates, 2'-O-methyl ribonucleotides, locked nucleic acid (LNA™), and peptide nucleic acids (PNAs). Unless specifically limited, the term encompasses nucleic acids containing known analogues of natural nucleotides that have similar binding properties as the reference nucleic acid. Unless otherwise indicated, a particular nucleic acid sequence also implicitly encompasses conservatively modified variants thereof (e.g., degenerate codon substitutions), alleles, orthologs, SNPs, and complementary sequences as well as the sequence explicitly indicated.The phrases "nucleic acid therapeutic", "therapeutic nucleic acid" and "TNA" are used interchangeably and refer to any modality of therapeutic using a nucleic acid as an active pharmaceutical ingredient of therapeutic agent to treat a disease or disorder.The term "pharmaceutically acceptable carrier" or "pharmaceutically acceptable excipient "includes any of the standard pharmaceutical carriers / excipients, such as a phosphate buffered saline solution, TRIS / sucrose buffer, water, emulsions such as an oil / water or water / oil, and various types of wetting agents. The term also encompasses any of the agents approved by a regulatory agency of the US Federal government or listed in the US Pharmacopeia for use in animals, including humans, as well as any carrier or diluent that does not cause significant irritation to a subject and does not abrogate the biological activity and properties of the administered compound.The term "subject" or "patient" refers to a human or animal, to whom treatment, including prophylactic treatment, with the therapeutic nucleic acid according to the present disclosure, is provided. Animals include mammals, birds and fish. Preferably, the animal is a mammal, e.g., primate, rodent, lagomorph, companion animal or livestock. Primates include humans, chimpanzees, cynomolgus monkeys, spider monkeys, and macaques, e.g., rhesus macaque. Rodents include mice, rats, and hamsters. Livestock include cows, horses, pigs, sheep and goats. Preferably, the subject is a human. A human subject can be of any age, gender, race or ethnic group.The terms "therapeutic amount", "therapeutically effective amount", "effective amount" "amount effective", or "pharmaceutically effective amount" of an active agent (e.g., a TNA described herein) are used interchangeably to refer to an amount that is sufficient to produce a desired effect, e.g., expression or inhibition of a target gene / sequence or disease modification. When the TNA is an mRNA, then an "effective amount" may be an amount sufficient to produce an increase in expression of a target polypeptide in comparison to the normal expression level, if any, detected in the absence of the messenger RNA. Suitable assays for measuring expression of a target gene or target sequence include, examination of protein or RNA levels using techniques known to those of skill in the art such as dot blots, northern blots, in situ hybridization, ELISA, immunoprecipitation, enzyme function, fluorescence or luminescence of suitable reporter proteins, as well as phenotypic assays. The terms include prophylactic or preventative amounts of an active agent is an amount sufficient to eliminate or reduce the risk, lessen the severity, or delay the onset of a disease, disorder or condition.The terms "dose" and "dosage" is the amount of an active agent administered at any given time. Dosage levels are based on a variety of factors, including the specific disease or disorder, the age, weight, sex, medical condition of the patient, the severity of the condition, the route of administration, and the particular active agent or agents employed. The dosage regimen can be determined routinely by a physician using standard methods.As used herein the term "therapeutic effect" refers to a consequence of treatment, the results of which are judged to be desirable, safely achieved, and beneficial. A therapeutic effect can include, directly or indirectly, the arrest, reduction, or elimination of a disease manifestation or progression.The terms "treat," "treating," and / or "treatment" may be therapeutic, prophylactic or palliative, and include abrogating, inhibiting, delaying, slowing or reversing the progression of a disease, disorder or condition; ameliorating clinical symptoms of a disease, disorder or condition; or preventing or reducing the appearance of clinical symptoms of a disease, disorder or condition.The term "alkyl" refers to saturated monovalent hydrocarbon radical. Alkyls may be linear or branched and may be optionally substituted. Examples include C1-20 alkyl, Cus alkyl, Cue alkyl, Ci-14 alkyl, C1-12 alkyl, C1-10 alkyl, C1-9 alkyl, C1-8 alkyl, C1-7 alkyl, C1-6 alkyl, C1-5 alkyl, C1-4 alkyl, and C1-C 3 alkyl. Examples further include methyl, ethyl, 1-propyl, 2-propyl, 1-butyl, 2- methyl-l-propyl, 2-butyl, 2-methyl-2-propyl, 1-pentyl, 2-pentyl, 3-pentyl, 2-methyl-2-butyl, 3-methyl-2-butyl, 3-methyl-l-butyl, 2-methyl-l-butyl, 1-hexyl, 2-hexyl, 3-hexyl, 2-methyl-2- pentyl, 3-methyl-2-pentyl, 4-methyl-2-pentyl, 3-methyl-3-pentyl, 2-methyl-3-pentyl, 2,3- dimethyl-2-butyl, 3,3-dimethyl-2-butyl, 1-heptyl, 1-octyl, and the like.The term "alkyl ester" refers to -R50-C(=O)O-R51or -R50-OC(=O)-R51, wherein R50is alkylene and R51is alkyl.The term "alkenyl ester" to -R52-C(=O)O-R53or -R52-OC(=O)-R53, wherein R52is alkylene or alkenylene and R53is alkyl or alkenyl, with the proviso that R52and / or R53contain at least one carbon-carbon double bond.The term "alkyl carbonate" refers to -R50-O-C(=O)O-R51or -R50-OC(=O)O-R51, wherein R50is alkylene and R51is alkyl.The term "alkenyl carbonate" to -R52-O-C(=O)O-R53or -R52-OC(=O)O-R53, wherein R52is alkylene or alkenylene and R53is alkyl or alkenyl, with the proviso that R52and / or R53contain at least one carbon-carbon double bond.The term "alkylene" refers to a saturated bivalent hydrocarbon radical. Alkylenes may be linear or branched and may be optionally substituted. Examples include C1-20alkylene groups, such as C1-20 alkylene, C1-18 alkylene, Cue alkylene, C1-14 alkylene, C1-12 alkylene, C1-10 alkylene, C1-9 alkylene, C1-8 alkylene, C1-7 alkylene, C1-6 alkylene, C1-5 alkylene, C1-4 alkylene, and C1-C 3 alkylene.The term "alkene" refers to a group consisting of at least two carbon atoms and at least one carbon-carbon double bond, and an "alkyne" moiety refers to a group consisting of at least two carbon atoms and at least one carbon-carbon triple bond. The alkyl moiety, whether saturated or unsaturated, may be branched, straight chain, or cyclic.The term "alkenyl" refers to a hydrocarbon radical with one or more carbon-carbon double bonds, including radicals having "cis" and "trans" orientations, or by an alternative nomenclature, "E" and "Z" orientations. Alkenyls may be linear or branched and may be optionally substituted. Examples include C2-20 alkenyl groups, such as, C2-20 alkenyl, C2-18 alkenyl, C2-16 alkenyl, C2-14 alkenyl, C2-12 alkenyl, C2-10 alkenyl, C2-9 alkenyl, C2-8 alkenyl, C2-7 alkenyl, C2-6 alkenyl, C2-5 alkenyl, C2-4 alkenyl, and C2-3 alkenyl."Alkenylene" refers to an aliphatic bivalent hydrocarbon radical with one or more carboncarbon double bonds, including radicals having "cis" and "trans" orientations, or by an alternative nomenclature, "Z" and "E" orientations, respectively. Alkenylenes may be linear or branched and may be optionally substituted. Examples include C2-20 alkenylene groups, such as, C2-20 alkenylene, C2-18 alkenylene, C2-16 alkenylene, C2-14 alkenylene, C2-12 alkenylene, C2-10 alkenylene, C2-9 alkenylene, C2-8 alkenylene, C2-7 alkenylene, C2-6 alkenylene, C2-5 alkenylene, C2-4 alkenylene, and C2-3 alkenylene."Alkynyl" refers to a hydrocarbon monovalent radical with one or more carbon-carbon triple bonds. "Alkynylene" refers to a hydrocarbon bivalent radical with one or more carboncarbon triple bonds (i.e., a group derived from an alkyne with two attachment points)."Aryl" refers to a monovalent aromatic group derived from an arene by removal of a hydrogen atom from a ring carbon atom. As used herein, aryl includes substituted or unsubstituted single-ring aromatic groups and polycyclic ring systems having two or morecyclic rings in which two or more carbons are common to two adjoining rings, wherein at least one of the rings is aromatic."Arylyene" or "arenediyl" refers to a bivalent aromatic group derived from an arene by removal of a hydrogen atom from two different ring carbon atoms, forming a group with two attachment points. In multi-ring systems, the two hydrogens may be removed from the same ring or different rings.Cyclic groups of the present invention include monocyclic and bicyclic groups (bridged or fused)."Carbocycle" and "carbocyclic" refers to a C3-C20 monocyclic or polycyclic (e.g., bicyclic or tricyclic), saturated, partially saturated or unsaturated ring(s), in which all the atoms composing the ring are carbon atoms. Ring moieties include fused, spirocyclic and bridged bicyclic rings. Saturated carbocyclic rings include, for example, "cycloalkyl" rings, e.g., cyclopropyl, cyclobutyl, etc. Carbocyclyl is a monovalent radical of a carbocycle, i.e., a carbocycle functional group with one attachment point. Carbocyclediyl or carbocyclene is a bivalent radical of a carbocycle, i.e., a carbocycle functional group with two attachment points."Cycloalkyl" refers to a monovalent saturated carbocyclic ring radical. Cycloalkyls may be optionally substituted. Cycloalkyl groups include groups having from 3 to 18 ring atoms. Cycloalkyl groups include, but are not limited to: cyclopropyl, cyclobutyl, cyclopentyl, cyclopentenyl, cyclohexyl, cyclohexenyl, cycloheptyl, cyclooctyl, cyclononyl, cyclodecyl and norbornyl."Cycloalkylene" as used herein refers to a bivalent, saturated, 3-18-membered carbocyclic ring radical with two attachment points. Specific monocyclic cycloalkylene groups include, but are not limited to, cyclopropylene, cyclobutylene, cyclopentylene, cyclohexylene, cycloheptylene, cyclooctylene, cyclononylene, cyclodecylene, cycloundecylene, cyclododecylene, and the like. In one embodiment, the cycloalkylene is cyclopropylene.The terms "include", "includes" and "including" are open-ended and not limited to examples provided."Halogen" refers to F, Cl, Br or I."Heterocycle" "heterocyclic" and "heterocyclic ring" are used interchangeably and refer to 5-20 aromatic or 3-20 aliphatic cyclic group having at least one ring heteroatom and at least one ring carbon atom. In one embodiment, the heteroatom is oxygen, sulfur, or nitrogen. A heterocycle containing more than one heteroatom may contain different heteroatoms. "Heterocyclyl" is a monovalent radical of a heterocycle and have one attachment point. An aromatic heterocyclyl is also referred to a "heteroaryl". Heterocyclene or heterocyclediyl refers to a bivalent heterocycle group with two attachment points. Heterocyclyl and heterocyclene moieties include both monocyclic and multicyclic (e.g., bicyclic or tricyclic)ring moieties. Ring moieties include fused, spirocyclic and bridged bicyclic rings and may comprise one or more heteroatoms in one or more of the rings. Either ring of a bicyclic heterocycle may be saturated, partially unsaturated or aromatic. The heterocycle may be attached to the rest of the molecule via a ring carbon atom or a ring nitrogen atom. Examples of heterocyclic functional groups include aziridinyl, diaziridinyl, thiaziridinyl, azetidinyl, diazetidinyl, triazetidinyl, thiadiazetidinyl, thiazetidinyl, pyrrolidinyl, pyrazolidinyl, imidazolinyl, isothiazolidinyl, thiazolidinyl, piperidinyl, piperazinyl, hexahydropyrimidinyl, azepanyl, and azocanyl."Heteroarene" is an aromatic compound derived from an arene by replacement of one or more methine (-C=) and / or vinylene (-CH=CH-) groups by trivalent or divalent heteroatoms, respectively."Heteroaryl" refers to a monovalent group with one attachment point derived from heteroarene by removal of a hydrogen atom from a ring atom."Heteroarylene" or "heteroarenediyl" refers to a bivalent group derived from a heteroarene by removal of a hydrogen atom from two different ring atoms, forming a group with two attachment points. The hydrogens may independently be removed from either a carbon or nitrogen atom (as available). In multi-ring systems, the two hydrogens may be removed from the same ring or different rings. Heteroaryls of the present invention include 5-18- membered aromatic radicals (e.g., C5-C13 heteroaryl), preferably 5-10-membered aromatic groups, that includes one or more ring heteroatoms selected from nitrogen, oxygen and sulfur, and which may be a monocyclic or polycyclic (e.g., a bicyclic, tricyclic or tetracyclic ring system). A polycyclic heteroaryl group may be fused or non-fused. The heteroatom(s) in the heteroaryl radical are optionally oxidized. One or more nitrogen atoms, if present, are optionally quaternized."Pharmaceutically acceptable salt" as used herein refers to pharmaceutically acceptable organic or inorganic salts of an ionizable lipid of the disclosure."Spirocycle" and "spiroheterocycle" refer is a 5- to 20-membered bicyclic ring system functional group, including spi ro [cycloa Ikyl] and spiro[cycloalkenyl] with both rings connected through a carbon single atom. A spirocycle / spiroheterocycle can be fully saturated or can be partially unsaturated. The rings can be different in size and nature, or identical in size and nature. Examples include spiropentanyl, spriohexanyl, spiroheptanyl, spirooctanyl, spirononanyl, or spirodecanyl. One or both of the rings in a spiro(hetero)cyclcle can be fused to another ring carbocyclic, heterocyclic, aromatic, or heteroaromatic ring. A (C5-C14) spirocycloalkyl, e.g., is a spirocycle containing between 5 and 14 carbon atoms. "Spiroheterocycloalkyl" or "spiroheterocyclyl" is understood to mean a spirocyclyl as defined above wherein at least one of the rings is a heterocycle, i.e., a ring containing a heteroatom. In one embodiment, the heteroatom is oxygen, sulfur, or nitrogen. "Amine" or "amino" as used herein interchangeably refers to a functional group that contains a basic nitrogen atom with a lone pair.The term "pharmaceutically acceptable salt" as used herein refers to pharmaceutically acceptable organic or inorganic salts of an ionizable lipid of the disclosure.Ranges set forth herein are inclusive of the end values.Hydrogen atoms connected to carbons may be substituted with deuterium (2H) atoms.Where two or more variables within a chain of variables are absent and designated as "a bond", the absent variables are, together, considered as one bond connecting the present adjacent variables. As an example, for the group -R62-R123-R63-R73-R74-R66, when R73 and R74 are each "a bond", then the groups is read as -R62-R123-R63-R66.Double bonds may be indicated as "=" or implicit based on valency rules.All patents and other publications; including literature references, issued patents, published patent applications, and co-pending patent applications; cited throughout this application are expressly incorporated herein by reference in their entireties.LipidLipids used in delivery of nucleic acids, e.g., mRNA include a head group and optional linker moiety and a hydrocarbon containing lipid tail. The head group is typically an amine- containing group when used as a typical ionizable lipid. An ionizable cationic lipid comprise a hydrophilic ionizable amine headgroup. Some cationic lipids have a permanently charged quaternary ammonium group as their head. Other ionizable cationic lipids have a pKa- dependent head group. This headgroup is responsible for condensing mRNA or other nucleic acids. The headgroup provides positive charge to interact with the negatively charged nucleic acids. Connecting the headgroup to the is an optional linker moiety which is preferably biodegradable for safe clearance after nucleic acid delivery. The headgroup is essential for self-assembly and phospholipid membrane fusion within the lipid nanoparticle. The headgroup-linker- is connected to a hydrophobic tail group. Ionizable cationic lipids typically have hydrophobic hydrocarbon chains as or as part(s) of the tail group. These chains promote self-assembly of the lipid nanoparticles. They also facilitate membrane fusion during cellular uptake. The hydrophobic tails further contribute to the overall stability of the lipid nanoparticle.In one aspect, the invention provides for a lipid as disclosed herein. The lipid may be used, e.g., in a lipid vesicle including liposomes, in a lipid film, and lipid nanoparticles (LNPs). The lipid may be a cationic lipid. The lipid may further be a ionizable cationic lipid.Lipid nanoparticlesIn another aspect, the invention provides for a lipid nanoparticle (LNP) comprising a lipid of the present invention. In some embodiments, the present disclosure provides for a LNP composition comprising a plurality of LNPs and at least one pharmaceutically acceptable carrier, diluent or excipient.The LNPs of the invention are lipid vesicles with a diameter that is typically in the range of 25-1000 nm. LNPs of the invention comprise multiple lipids, at least one of which is positively charged (cationic) at low pH (enabling RNA complexation and endosomal escape).The cationic lipid is preferably an ionizable cationic lipid that is substantially in the neutral form in an LNP at physiological pH. In addition to a (ionizable) cationic lipid, the LNP may further comprises a non-cationic, structural / helper lipid, a sterol (to provide membrane fluidity) and a polymer conjugated lipid (to prevent aggregation).LNPs of the invention may comprise a targeting moiety, such as a protein or peptide or a cluster of peptides and / or small molecule targeting ligands.LNPs of the invention may comprise a labelling moiety, such as a fluorophore small molecule (BODIPY and the like) for tracking purposes with confocal microscopy and fluorescence imaging methods.LNPs of the invention may further comprise a diagnostic or therapeutic agent and be used to deliver the agent, such as TNA, to a cell, tissue or organ. In some embodiments, the LNP comprises a therapeutic agent such as a TNA (e.g., mRNA), protein, peptide or other sensitive cargo encapsulated or contained in the lipid portion of the particle, thereby protecting it from enzymatic degradation, excretion or immunogenic or other reaction.LNP ComponentsAccording to some embodiments, the lipid particles of the disclosure have a mean diameter of: from about 25 nm to about 125 nm, about 50 nm to about 100 nm, about 100 nm to about 200 nm, about 200 nm to about 300 nm, from about 300 nm to about 400 nm, from about 400 nm to about 500 nm, from about 500 nm to about 750 nm, from about 750 nm to about 1000 nm, from about 50 nm to about 500 nm, from about 25 nm to about 1000 nm, less than about 1000 nm, less than about 750 nm, less than about 500 nm, less than about 250 nm, less than about 150 nm, less than about 100 nm, less than about 75 nm, less than about 50 nm, less than about 40 nm, less than about 30 nm, less than about 25 nm, or less than about 20 nm. Lipid particle (e.g., lipid nanoparticle) size can be determined, e.g., by quasi-elastic light scattering using a Malvern Zetasizer Nano ZS (Malvern, UK) or Wyatt Dynapro DLS (Wyatt Technologies, Santa Barbara CA).In some embodiments, the LNPs may be relatively homogenous. A polydispersity index may be used to indicate the homogeneity of the LNPs. A small, for example less than 0.3 or less than 0.2, polydispersity index generally indicates a narrow particle size distribution. A composition of the LNPs described herein may have a polydispersity index from about 0 to about 0.25 or to about 0.30, such as 0.01, 0.02, 0.03, 0.04, 0.05, 0.06, 0.07, 0.08, 0.09, 0.10, 0.11, 0.12, 0.13, 0.14, 0.15, 0.16, 0.17, 0.18, 0.19, 0.20, 0.21, 0.22, 0.23, 0.24, 0.25, 0.26, 0.27, 0.28, 0.29 or 0.30. In some embodiments, the polydispersity index of the LNP composition may be from about 0 to about 0.30 or 0.05 to 0.20.In one aspect, the LNP comprises: a (ionizable) cationic lipid, a sterol or a derivative thereof, a non-cationic lipid, and a polymer-conjugated lipid. In one embodiment, the LNP comprises more than one (ionizable) cationic lipid, more than one sterol or a derivative thereof, more than one non-cationic lipid, and / or more than one polymer-conjugated lipid. In another embodiment, the lipid particle (e.g., lipid nanoparticle) comprises a cationic lipid, a noncationic phospholipid, cholesterol and a PEGylated lipid (polymer conjugated lipid). In a further embodiment, the cationic lipid, non-cationic phospholipid, cholesterol and a PEGylated lipid are present in a molar ratio of about 50:7:40:3. 50: 10:38.5: 1.5, respectively. In one embodiment, of the total lipid content, the LNP comprises about: 40-50 mol%, 45-50 mol%, 50-55 mol%, 45-46 mol%, 46-47 mol%, 47-48 mol%, 48-49 mol%, or 49- 50 mol%, 45 mol%, 45.5 mol%, 46 mol%, 46.5 mol%, 47 mol%, 47.5 mol%, 48 mol%, 48.5 mol%, 49 mol%, or 49.5 mol% (ionizable) cationic lipid. In one embodiment of the total lipid content, the LNP comprises about: 5-25 mol%, 5-15 mol%, 10-12 mol%, 5-6 mol%, 6-7 mol%, 7-8 mol%, 8-9 mol%, 9-10 mol%, 10-11 mol%, 11-12 mol%, 12-13 mol%, 13-14 mol%, or 14- 15 mol% non-cationic lipid. In one embodiment of the total lipid content, the LNP comprises about: 25-55 mol%, 30-45 mol%, 35-40 mol%, 30-31 mol%, 31-32 mol%, 32-33 mol%, 33-34 mol%, 35-35 mol%, 35-36 mol%, 36-37 mol%, 38-38 mol%, 38-39 mol%, or 39-40 mol% sterol. In one embodiment of the total lipid content, the LNP comprises about: 0.5-15 mol%, 1-5 mol%, 1-3 mol%, 1.5-2.5 mol%, 1-2 mol%, 2-3 mol%, 3-4 mol%, or 4-5 mol% polymer conjugated lipid, e.g., PEGylated lipid. In one embodiment the lipid nanoparticle comprises a total lipid content that is 20-60 mol% (ionizable) cationic lipid, 5-25 mol% non-cationic lipid, 25-55 mol% sterol, and 0.5-15 mol% polymer conjugated lipid, e.g., PEGylated lipid. In one embodiment, the lipid nanoparticle comprises a total lipid content that is 40-50 mol% (ionizable) cationic lipid, 5-15 mol% non-cationic lipid, 30-45 mol% sterol, and 1-5 mol% polymer conjugated lipid, e.g., PEGylated lipid. In one embodiment, the lipid nanoparticle comprises a total lipid content that is 45-50 mol% (ionizable) cationic lipid, 10-12 mol% noncationic lipid, 35-40 mol% sterol, and 1-3 mol% polymer conjugated lipid, e.g., PEGylated lipid. In one embodiment, the lipid nanoparticle comprises a total lipid content that is 45-50 mol% (ionizable) cationic lipid, 10-12 mol% non-cationic lipid, 35-40 mol% sterol, and 1.5-2.5 mol% polymer conjugated lipid, e.g., PEGylated lipid conjugate.Cationic and ionizable lipidsIn some aspects, the lipid nanoparticle of the present invention comprises a cationic lipid. In some aspects, the cationic lipid is an ionizable cationic lipid. The ionizable cationic lipid is positively charged at low pH, which facilitates association with the negatively charged nucleic acid. The ionizable cationic lipid is neutral at physiological pH (pH 7.4). The ability of these lipids to ionize at low pH aids in endosomal escape of the nucleic acid into the cytoplasm.In one aspect, the invention provides for a pharmaceutical composition comprising a lipid nanoparticle, wherein the lipid nanoparticle comprises a cationic lipid. In someembodiments, the cationic lipid is an ionizable cationic lipid. Such LNPs can be used to deliver a diagnostic or therapeutic agent to a target cell, tissue or organ in a subject.Exemplary ionizable cationic lipids are described in International PCT patent publications WO2022 / 246571, W02018 / 011633, WO2017 / 117528, WO2017 / 099823, WO2017 / 075531, WO2017 / 049245, W02017 / 004143, W02016 / 081029, WO2015 / 199952, WO2015 / 095346, W02015 / 095340, W02015 / 074085, WO2015 / 061467, WO2013 / 148541, WO2013 / 126803, WO2013 / 116126, W02013 / 089151, WO2013 / 086373, WO2013 / 086354, WO2013 / 086322, WO2013 / 049328, WO2013 / 033563, W02013 / 016058, W02013 / 006825, W02012 / 162210, WO2012 / 099755, WO2012 / 054365, WO2012 / 044638, W02012 / 040184, W02012 / 031043, W02012 / 016184, W02012 / 000104, W02011 / 153120, W02011 / 141705, W02011 / 141704, W02011 / 090965, W02011 / 071860, W02011 / 066651, W02011 / 038160, W02011 / 022460, W02011 / 000107, W02011 / 000106, W02010 / 144740 , W02010 / 129709, W02010 / 088537, W02010 / 054406 , W02010 / 054405, W02010 / 054401, W02010 / 054384, W02010 / 048536, W02010 / 042877, W02009 / 132131, W02009 / 127060, W02009 / 086558, W02008 / 042973, W02006 / 069782, W02006 / 007712, WO2005 / 121348, and W02005 / 120152; and US patent publications US2018 / 0028664, US2018 / 0005363, US2017 / 0119904, US2017 / 0210697, US2016 / 0376224, US2016 / 0317458, US2016 / 0311759, US2016 / 0151284, US2015 / 0376115, US2015 / 0239926, US2015 / 0203446, US2015 / 0141678, US2015 / 0140070, US2015 / 0064242, US2015 / 0057373, US2014 / 0308304, US2014 / 0255472, US2014 / 0200257, US2014 / 0141070, US2014 / 0039032, US2013 / 0338210, US2013 / 0323269, US2013 / 0303587, US2013 / 0274523, US2013 / 0274504, US2013 / 0274504, US2013 / 0202684, US2013 / 0195920, US2013 / 0178541, US2013 / 0123338, US2013 / 0116307, US2013 / 0090372, US2013 / 0065939, US2012 / 0202871, US2012 / 0149894, US2012 / 0128760, US2012 / 0101148, US2012 / 0058144, US2012 / 0027796, US2010 / 0324120, US2010 / 0130588, US2010 / 0062967, US201 / 0256175, US201 / 0117125, US201 / 0076335, US2009 / 0023673, US2006 / 0083780, US2006 / 0051405, US2006 / 0008910, and US2003 / 0022649, each of which are incorporated by reference in their entireties.Further examples include 3-(didodecylamino)-Nl,Nl,4-tridodecyl-l-piperazineethanamine (KL1O), Nl-[2-(didodecylamino)ethyl]-Nl,N4,N4-tridodecyl-l,4- piperazinediethanamine (KL22), 14,25-ditridecyl-15, 18,21 ,24-tetraaza-octatriacontane (KL25), 1.2-dilinoleyloxy-N,N- dimethylaminopropane (DLin-DMA), 2,2-dilinoleyl-4-dimethylaminomethyl-[l,3]-dioxolane (DLin-K-DMA), (6Z,9Z,28Z,31Z)-heptatriacont-6,9,28,31-tetraene-19-yl 4- (dimethylamino)butanoate (DLin-MC3-DMA), 2.2-dilinoleyl-4-(2-dimethylaminoethyl)-[l,3]- dioxolane (DLin-KC2-DMA), l,2-dioleyloxy-N,N-dimethylaminopropane (DODMA), 2-({8- [(3P)-cholest-5-en-3-yloxy]octyl}oxy)-N,N-dimethyl-3-[(9Z,12Z)- octadeca-9,12-dien-l- yloxy]propan-l -amine (Octyl-CLinDMA), (2R)-2-({8-[(3P)-cholest-5-en-3-yloxy]octyl}oxy)-N,N- dimethyl-3-[(9Z,12Z)-octadeca-9,12-dien-l-yloxy]propan-l-amine (Octyl-CLinDMA (2R)), (2S)-2-({8-[(3P)-cholest-5-en-3-yloxy]octyl}oxy)-N,N-dimethyl-3-[(9Z,12Z)-octadeca-9,12- dien-l-yloxy]propan-l-amine (Octyl-CLinDMA (2S)), ((4-hydroxybutyl)azanediyl)bis(hexane- 6,l-diyl)bis(2-hexyldecanoate)), 3-((4,4-bis(octyloxy)butanoyl)oxy)-2-((((3-(diethylamino)propoxy)carbonyl)oxy)methyl)propyl (9Z,12Z)-octadeca-9,12-dienoate, and 8- [(2-hydroxyethyl)[6-oxo-6-(undecyloxy)hexyl]amino]-octanoic acid, 1-octylnonyl ester.In one aspect, the ionizable cationic lipid has a structural formula selected from the group consisting of: Compound (1):(2),Compound (3):(4); or a pharmaceutically acceptable salt or co-crystal thereof, wherein:R1for each occurrence is independently selected from: -H, optionally substituted C1-C6 alkyl, -[CFhJi-eOH, optionally substituted C2-C6 alkenyl, -C(=O)CH2N(R72)2, -P(O)(OR17)(OR18), - P(O)(OR17)(NR19), or -P(O)(NR19)(NR20);R3is selected from: -H, optionally substituted C1-C6 alkyl, -[CH2]i-eOH, optionally substituted C2-C6alkenyl, -P(O)(OR17)(OR18), -P(O)(OR17)(NR19), -P(O)(NR19)(NR20) or R4;R2is selected from: is absent (i.e., a lone pair on N), -H or optionally substituted C1-C6 alkyl; provided that when R2is -H or optionally substituted C1-C6 alkyl, the nitrogen atom, to which R1, R2, and R3are all bonded to, is protonated;R4for each occurrence is independently selected from:R55for each occurrence, is independently selected from:A1, A3, A4, A5, A6, A7, A8, A12and A13, for each occurrence, are each independently selected5 from: a bond, -O-, -C(=O)-, -C(=S)-, -OC(=O)-, -C(=O)O-, -OC(=O)O-, -OC(=S)-, -C(=S)O-, -OC(=S)O-,-S-C(=O)-, -C(=O)-S-, -S-S-, -S(OH)-, -S(=O)-, -S(=O)2-, -S(OH)2-, -C(=O)N(R14)-, - N(R14)C(=O)-, -N(R14)C(=O)N(R14)-, -O-C(=O)C(R14)2C(=O)O-, -C(=O)O-C(R14)2C(=O)O-, -O- C(=O)C(R14)2-O-C(=O)-, -O-C(R14)2C(=O)O-, -O-C(=O)C(R14)2C-O-, -O-C(R14)2-O-C(=O)-, -C(=O)- O-C(R14)2C-O-, -O-C(=O)-O-R41-O-C(=O)O-, -0-C(=0)-[CH2]O-4-C(=0)-0-, -P(OH)-, -P(OH)( R14)-, - P(=O)(OH)-, -P(O R14)2-, -P(O R14)- or -P(=O)(O R14)-, -CH(R14)-, -C(R14)2-, -CH(OH)-, -CH(O R14)-, -CH(NH2)-, -CH(NH[R14])-, -C(NH[OH])-, -C(NH[O R14])-, -N(R14)-, -S-, -C(=O)C(R14)2C(=O)-, - C(R14)2C(=O)-, -C(=O)C(R14)2-O-, -C(R14)2C(R14)2-, -C(R25)2-O-, -O-C(R14)2C(R14)2-, -O- C(R14)2C(=O)-, -C(=O)-O- R41-O-C(=O)-;R41for each occurrence, are each independently an optionally substituted group selected from: a bond, linear or branched Ci-Cs alkylene, Ci-Cs alkenylene or Ci-Cs alkynylene; C3-C10 heterocyclylene or C3-C10 carbocyclylene; or -R114-C3-C6-carbocyclylene-R115-, -R114-C3-Ce- heterocyclylene-R115-, -R114-C3-C6-cycloakylene-R115- -R114-CH=CH-CH2-CH=CH-R115- , -R114- CH=CH-CH2-cPr-R115-, -R114-cPr-CH2-CH=CH-R115-, -R114-cPr-CH2-cPr-R115-, -R114-CH=CH-CD2- CH=CH-R115-, -R114-CH=CH-CD2-cPr-R115-, -R114-cPr-CD2-CH=CH-R115- or -R114-cPr-CD2-cPr-R115- ; wherein, R114and R115are each independently selected from: a bond or linear or branched C1-C6 alkylene, C1-C6 alkenylene or C1-C6 alkynylene; wherein -cPr- is cyclopropylene (cyclopropane-1,2 diyl); each double bond or cPr has a cis configuration; and D denotes deuterium (2H);A2for each occurrence, is independently selected from: a bond, -O-, -OC(=O)-, -C(=O)O-, - OC(=O)O-, -S-C(=O)-, -C(=O)-S-, -S-S-, -C(=O)N(R14)-, -N(R14)C(=O)-, -N(R14)C(=O)N(R14)-, -O- C(=O)C(R14)2C(=O)O-, -C(=O)O-C(R14)2C(=O)O-, -O-C(=O)C(R14)2-O-C(=O)-, -O-C(R14)2C(=O)O-, - O-C(=O)C(R14)2C-O-, -O-C(R14)2-O-C(=O)-, -C(=O)-O-C(R14)2C-O- or -O-C(=O)-O-R41-O-C(=O)O-;R6for each occurrence, is independently selected from: a bond, -R58-N(R42)-R58-, -R58- CH(R42)-R58-, C1-C12alkylene or C2-C12alkenylene;R7for each occurrence, is independently selected from: a bond (i.e., when present A2is bonded directly toptionally substituted C1-C18 alkylene, or optionally substituted C2-C18alkenylene;R8and R9, for each occurrence, are each independently selected from:R42;R12for each occurrence, is independently selected from: -H, optionally substituted C1-C16 alkyl or optionally substituted C2-C16alkenyl;R13for each occurrence, is independently selected from: optionally substituted C1-C16 alkyl or optionally substituted C2-C16 alkenyl;R14, for each occurrence, is independently selected from: -H, optionally substituted C1-C10 alkyl, optionally substituted C1-C10 alkenyl or optionally substituted C1-C10 alkynyl;R15for each occurrence, is independently selected from: a bond, -R41-N(R42)-R58-, -R41- CH(R42)-R58-, or C1-C6 alkylene;R16for each occurrence, is independently selected from: a bond or — [CH2]k-;R17and R18are each independently selected from: -H, C1-C5 alkyl, an alkali metal cation, an alkaline earth metal cation, ammonium cation, methyl ammonium cation, or a pharmaceutically acceptable base;R19and R20, for each occurrence, are each independently selected from: -OH, formyl, acetyl, pivaloyl, -NH2, -NH(CH3), -NH(CH2CH3), -N(CH3)2, -NHC(=O)H, -NHC(=O)CH3, or C3-C5-alkyl;R29through R34, for each occurrence, are each independently selected from: -CH2-, -NH-, -S-, or -O-;R35for each occurrence, is independently selected from: -OH or C1-C6 acyl; optionally substituted C1-C6 alkyl, C1-C6 alkyl ester, C1-C6 alkyl ether or C1-C6 alkyl carbonate;R36and R37, are each independently selected from: -H, -C(=O)C1-C6 alkyl, -C(=O)C2-C6 alkenyl, -C(=O)-O-C1-C6alkyl, or -C(=O)-O-C2-C6alkenyl, C1-C6 alkyl, or C2-C6alkenyl;R38is selected from: -H, C1-C24 alkylene or C1-C24 alkenylene;R41for each occurrence, is independently selected from: a bond or C1-C5 alkylene;R42for each occurrence, is independently selected from: C6-C24 alkyl, C6-C24 alkyl carbonate, C6-C24 alkyl ether, C6-C24 alkyl ester, -R58-A1-R59or -R58-A1-R59-A2-R60, wherein R42has between 6-24 total carbon atoms;R58R59, and R60, for each occurrence, are each independently selected from: a bond, -O-, -O- C(=O)-, -OC(=O)O-, C1-C24 alkylene or C2-C24 alkenylene;R62, R63, R64, R67, R68, R69, R74, R75, R76, R77, R108, R109, R110and R111for each occurrence, are each independently selected from: a bond, C1-C10 alkylene, C1-C10 alkenylene, C1-C10 alkynylene, C3-C7cycloalkylene, C3-C7cycloalkenylene, C5-C10-spirocycloalkylene, C3-C10- carbocyclylene, C3-Ci0-heterocyclylene, ,-R114-C3-C6-cycloalkylene-R115-, -R114-CH=CH-CH2- CH=CH-R115- , -R114-CH=CH-CH2-cPr-R115-, -R114-cPr-CH2-CH=CH-R115-, -R114-cPr-CH2-cPr-R115-, - R114-CH=CH-CD2-CH=CH-R115-, -R114-CH=CH-CD2-cPr-R115-, -R114-cPr-CD2-CH=CH-R115- or -R114- cPr-CD2-cPr-R115-; wherein, R114and R115are each independently selected from: a bond, C1- C10 alkylene, C1-C10 alkenylene or C1-C10 alkynylene; -cPr- is cyclopropylene (cyclopropane- 1,2 diyl); each double bond or cPr has a cis configuration; and D denotes deuterium (2H);R114and R115for each occurrence, are each independently, selected from: a bond, C1-C10 alkylene, C1-C10 alkenylene or C1-C10 alkynylene; -cPr- is cyclopropylene (cyclopropane-1,2 diyl); each double bond or cPr has a cis configuration; and D denotes deuterium (2H);R61, R73and R107, for each occurrence, are each independently selected from:R116, for each occurrence, is independently selected from: -H, -F, -CF3, -Cl, -Br, -I, -CH3, - CH2R14, -CHR142, -CR143, -OH, -OR14, -NH2, -NHR14, -N(R14)2, -SH, -SR14, -S(=O)H, -S(=O)R14, - S(=O)2H, -S(=O)2R14, -PH2, -PHR14, -PR142, -P(OH)2, -POHR14, -PR142, -P(OR14)2, -P(O)H2, - P(O)HR14, -P(O)R142, -P(O)(OH)2, -P(O)(OR14)OH, -P(O)(OR14)2, -SiH3, -Si H2R14, -SiH R142or - SiR143;R65, R70, R78, R105and R112, for each occurrence, are each independently selected from: a bond, C1-C10 alkylene, C1-C10 alkenylene, C1-C10 alkynylene, C3-C8cycloalkylene, C3-C8cycloalkenylene, C5-C10-spirocycloalkylene, C3-C10-carbocyclylene, C3-C10-heterocyclylene, - CH=CH-CH2-CH=CH- , C3-C6-cycloakylene, -CH=CH-CH2-cPr-, -cPr-CH2-CH=CH-, -cPr-CH2-cPr-, - CH=CH-CD2-CH=CH- , -CH=CH-CD2-cPr-, -cPr-CD2-CH=CH- -or -cPr-CD2-cPr-, where -cPr- is cyclopropylene, each double bond or cPr has a cis configuration and D denotes deuterium (2H);R66, R71, R79, R106and R113, for each occurrence, are each independently selected from: -H, linear or branched C1-C10 alkyl, C1-C10 alkenyl, or C1-C10 alkynyl, C5-C10-spirocycloalkyl, C3-C10- carbocyclyl, C3-C10-heterocyclyl, optionally substituted aryl, optionally substituted heteroaryl, C3-C18-heterocycloalkyl, optionally substituted aryl, optionally substituted heteroaryl, -R26-CH=CH-CH2-CH=CH-R28, -R26-CH=CH-CH2-cPr-R28, -R26-cPr-CH2-CH=CH-R28, - R26-cPr-CH2-cPr-R28, -R26-CH=CH-CD2-CH=CH-R28, -R26-CH=CH-CD2-cPr-R28, -R26-cPr-cPr-R28, - R26-cPr-cBu-R28, -R26-cBu-cBu-R28, -R26-cBu-cPr-R28, -R26-cHx-cBu-R28, -R26-cHx-cBu-R28, -R26- cHx-cBu-R28, -R26-cPr-cHx-R28, -R26-cBu-cHxR28, -R26-cPr-R26-cPr-R28, -R26-cPr-R26-cBu-R28, -R26- cBu-R26-cBu-R28, -R26-cBu-R26-cPr-R28, -R26-cHx-R26-cBu-R28, -R26-cHx-R26-cBu-R28, -R26-cHx- R26-cBu-R28, -R26-cPr-R26-cHx-R28, -R26-cBu-R26-cHxR28, -R26-c-CD2-cPr-R28, -R26-cPr-CD2-cPr- R28, -R26-cPr-CD2-cPr-R28, -R26-cPr-CD2-CH=CH-R28, -R26-cPr-CD2-cPr-R28or -R26-C3-C6- cycloakylene-R26-C3-C6cycloakylene-R28; wherein, R26for each occurrence, is independently selected from: a bond, C1-C10 alkylene, C1-C10 alkenylene or C1-C10 alkynylene; and R28for each occurrence, is independently selected from: -H, C1-C10 alkyl, C1-C10 alkenyl or C1-C10 alkynyl; -cPr- is cyclopropylene (cyclopropane-1,2 diyl); -cBu- is cyclobutylene; -cHx- is cyclohexylene; each double bond or -cPr- has a cis configuration; and D denotes deuterium (2H);R72, for each occurrence, is independently selected from -H, optionally substituted C1-C6 alkyl, -[CH2]1-6OH or optionally substituted C2-C6 alkenyl;R96, for each occurrence, is independently selected from: -OH, -O-C1-C4 alkyl, -O-C(=O)-O-C1- C4 alkyl or -O-C(=O)-C1-C5 alkyl;R118and R119are independently selected from R1or R55; n is an integer selected from: 1, 2, 3, 4, 5 or 6; and k is an integer selected from: 1, 2, 3, 4, 5 or 6.In some embodiments, said carbocyclyl or heterocyclyl is independently an optionally substituted group selected from: cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, cycloheptyl, cyclopropenyl, cyclobutenyl, cyclopentenyl, cyclohexenyl, cycloheptenyl, cyclobutadienyl, cyclopentadienyl, cyclohexadienyl, cycloheptadienyl or cycloheptatrienyl.In some embodiments, said carbocyclyl or heterocyclyl is independently an optionally substituted group selected from: lH-pyrrolizidinyl, 1,2-dihydroquinolinyl, 1,5- naphthyridinyl, 1,8-naphthyridinyl, lH-indazolyl, lH-isochromenyl, lH-pyrrolizidinyl, 1- naphthyl, 2H-benzo[b][l,4]oxazinyl, 2H-benzo[e][l,2]oxazinyl, 2h-chromenyl, 2-naphthyl, 4H-quinolizinyl, adeninyl, azaindazolyl, azaindolyl, benzimidazolyl, benzo[b]thiophenyl, benzo[c][l,2,5]thiadiazolyl, benzo[c]isothiazolyl, benzo[c]thiophenyl, benzo[d]isothiazolyl, benzo[d]isoxazolyl, benzo[d]oxazolyl, benzo[d]thiazolyl, benzofuryl, benzyl, cinnolinyl, cumenyl, decahydroisoquinolinyl, decahydroquinolinyl, guaninyl, indazolyl, indenyl, indolyl, indolinyl, indolizinyl, isobenzofuran, isoindolyl, isoquinolinyl, phenyl, phthalazinyl, pteridinyl, purinyl, pyrido[2,3-b]pyrazinyl, pyrido[4,3-d]pyrimidinyl, pyrimidinyl, quinazolinyl, quinolinyl, quinoxalinyl, tetrahydroquinolinyl, tolyl or xylyl.In some embodiments, said carbocyclyl or heterocyclyl is independently an optionally substituted group selected from: furyl, imidazolyl, isothiazolyl, isoxazolyl, oxadiazolyl, oxazolyl, pyrrolyl, pyrazolyl, pyridazinyl, pyridyl (pyridinyl), pyrimidinyl, thiadiazolyl, thienyl, tetrazolyl, thiazolyl, triazolyl, , azepinyl, azetidinyl, dioxothiomorpholinyl, imidazolidinyl, morpholinyl, oxanyl, oxazinyl, oxazolidinyl, oxepinyl, oxetanyl, piperazinyl, piperidinyl, pyranyl, pyrrolidinyl, tetrahydrofuranyl, tetrahydropyranyl, thianyl, thiomorpholinyl or thiopyranyl.In some embodiments, said carbocyclyl or heterocyclyl is independently an optionally substituted group selected from: adamantanyl, azabicyclo[3.1.0]hexanyl, 3- azabicyclo[3.1.1]heptanyl, bicyclo[2.1.1]hexyl, bicyclo[2.2.1]heptyl, bicyclo[2.2.1]heptenyl, 3-oxa-8-azabicyclo[3.2.1]octanyl, 6-oxa-3-azabicyclo[3.1.1]heptanyl, 8-Methyl-8- azabicyclo[3.2.1]octanyl, 8-oxa-3-azabicyclo[3.2.1]octanyl, 3-oxa-6- azabicyclo[3.1.1]heptanyl, tricyclo[2.2.1.0(2,6)]heptanyl, 6,6-dimethylbicyclo[3.1.1]heptyl, or 2,6,6-trimethylbicyclo[3.1.1]heptyl, 5-azaspiro[2.3]hexanyl, 2-azaspiro[3.3]heptanyl, 2-oxa-6-azaspiro[3.3]heptanyl, spiro[2.2]pentanyl, spiro[3.3]heptanyl, spiro[2.3]hexanyl or spiro[2.5]octanyl, spiro[4.5]decanyl.In some embodiments, said carbocyclyl or heterocyclyl is independently an optionally substituted group selected from:In some embodiments, said carbocyclylene or heterocyclylene is independently an optionally substituted group selected from: furandiyl, imidazolediyl, isothiazolediyl, isoxazolediyl, oxadiazolediyl, oxazolediyl, pyrazolediyl, pyrrolediyl, pyridazinediyl, pyridinediyl, pyrimidinediyl, thiadiazolediyl, thiendiyl, tetrazolediyl, thiazolediyl, triazolediyl, azepinediyl, azetidinediyl, dioxothiomorpholinediyl, imidazolidinediyl, morpholinediyl, oxanediyl, oxazinediyl, oxazolidinediyl, oxepinediyl, oxetanediyl, piperazinediyl, piperidinediyl, pyranyl, pyrrolidindiyl, tetrahydrofurandiyl, tetrahydropyrandiyl, thianediyl, thiomorpholinediyl or thiopyrandiyl.In some embodiments, said carbocyclylene or heterocyclylene is independently an optionally substituted group selected from: cyclopropylene (cyclopropanediyl), cyclobutylene (cyclobutanediyl), cyclopentylene (cylcopentanediyl), cyclohexylene (cyclohexenediyl), cycloheptylene (cycloheptanediyl), cyclopropenediyl, cyclobutenylenediyl, cyclopentenylenediyl, cyclohexenediyl, cycloheptenediyl, cyclobutadienediyl, cyclopentadienediyl, cyclohexadienediyl or cycloheptadienediyl, cycloheptatrienediyl.In some embodiments, said carbocyclylene or heterocyclylene is independently an optionally substituted group selected from: adamantanediyl, azabicyclo[3.1.0]hexanediyl, 3- azabicyclo[3.1.1]heptanediyl, bicyclo[2.1.1] hexyl, bicyclo[2.2.1]heptyl,bicyclo[2.2.1]heptenyl, 3-oxa-8-azabicyclo[3.2.1]octanediyl, 6-oxa-3- azabicyclo[3.1.1]heptanediyl, 8-oxa-3-azabicyclo[3.2.1]octanediyl, 3-oxa-6- azabicyclo[3.1.1]heptanediyl, tricyclo[2.2.1.0(2,6)]heptanyl, 6,6- dimethylbicyclo[3.1.1]heptyl, or 2,6,6-trimethylbicyclo[3.1.1]heptyl, , 5- azaspiro[2.3]hexanediyl, 2-azaspiro[3.3]heptanediyl, 2-oxa-6-azaspiro[3.3]heptanediyl, spiro[2.2]pentanyl, spiro[3.3]heptanyl, spiro[2.3]hexanediyl, spiro[2.5]octanediyl or spiro[4.5]decanediyl.In some embodiments, said cycloalkylene, in each instance, is independently selected from:In some embodiments for any one or more variable selected from R3, R4, Rll, R12, R15, R16, R17, R18, R62, R63, R64, R65, R67, R68, R69, R70, R74, R75, R76, R77, R78, R105, R108, R109, R110, Rill, R112, R115 or R191, said carbocyclylene or heterocyclylene is independently an optionally substituted group selected from:In some embodiments, said carbocyclylene or heterocvclvlene is independently an optionallysubstituted group selected from:; wherein, R117 is selected fromIn some embodiments, for any one of Compounds (l)-(4), -R62-A3-R63-A4-R64-R65-R66, -R62-A3-R63-A4-R64-R73-R74_A7_R75_R65_R6^ _R62_A3_R63_A4_R64_R73_R76_A8_R77_R78_R79;_R67_A5_R68_A6_R69_R7O-R71, -R67_A5.R68.A6.R69.R1O7.R1O8.A12.R1O9.R1O5.R1O6 _R67_A5_R68_A6_R69_R107_R110_A13_R111_R112-R113, -R62-A3-R63-A4-R64-R65-R66, -R62-A3-R63-A4-R64-R74-A7-R75-R65-R66, -R62-A3-R63-A4-R64-R76-A8-R77-R78_R79;-R67-A5-R68_A6_R69_R7O_R71;_R67_A5_R68_A6_R69_R108_A12_R109_R105_R106A N D_R67-A5-R68-A6-R69-R110-A13-R111-R112-R113, for each occurrence, independently has: a) a total number of carbon atoms selected from: 10-24, 12-22, 14-22, 16-20, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23 or 24 carbon atoms; b) a longest linear chain (i.e., excluding side groups) with a total number of carbon atoms that is selected from: 10-24, 12-22, 14-22, 16-20, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23 or 24 carbon atoms. Note that the longest linear chain may contain non-carbon atoms including oxygen, nitrogen, sulfur and silicon, however, once the longest chain is identified, only the carbon atoms are counted; or c) .a longest linear chain (i.e., excluding side groups) with a total number of atoms (including non-carbon atoms) that is selected from: 10-24, 12-22, 14-22, 16-20, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23 or 24 atoms (including non-carbon atoms); wherein each variable, for each occurrence, is independently as described above for any one of Compounds (1) - (4) or for any embodiment of any compound herein.For clarity, the "longest liner chain" refers only to those atoms that directly form the chain, such that no bond between any two chain atoms can be broken without breaking the chain.In some embodiments, for any one of Compounds (l)-(4), R55, for each occurrence, is independently selected from:R62-A3-R66, -R62-A3-R63-R66, -R62-A3-R63-A4-R66, -A5-R68-A6-R69-R70-R71, -R62-A3-R63-A4-R64-R66A4_R64_R74_A7_R75_R66; Qr-R62-A3-R63-A4-R64-R74-A7-R75-R65-R66;; wherein each variable, for each occurrence, is independently as described above for any one of Compounds (1) - (4) or for any embodiment of any compound herein. In some further embodiments, said R55said group has: a) a total number of carbon atoms selected from: 10-24, 12-22, 14-22, 16-20, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23 or 24 carbon atoms; or b) a longest linear chain (i.e., excluding side groups) with a total number of carbon atoms or total number of atoms (i.e., carbon and heteroatoms) that is selected from: 10-24, 12-22, 14-22, 16-20, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23 or 24 carbon atoms.In some embodiments, for any one of Compounds (l)-(4), said compound comprises two R55groups independently selected from:-R62-A:-R66, -R62-A3-R63-R66, -R62-A3-R63-A4-R66, -A5-R68-A6-R69-R70-R71, -R62-A3-R63-A4-R64-R66, -R62-A3-R63-A4-R64-R74-R66, -R62-A3-R63-A4-R64-R74-R66, -R62-A3-R63-A4-R64-R74-A7-R66, -R62-A3-R63A4-R64-R74-A7-R75-R66, or -R62-A3-R63-A4-R64-R74-A7-R75-R65-R66; wherein, each variable, for each occurrence, is independently as described above for any one of Compounds (1) - (4) or for any embodiment of any compound herein. In some further embodiments, said R55said group has: a) a total number of carbon atoms selected from: 10-24, 12-22, 14-22, 16-20, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23 or 24 carbon atoms; or b) a longest linear chain (i.e., excluding side groups) with a total number of carbon atoms or total number of atoms (i.e., carbon and heteroatoms) that is selected from: 10-24, 12-22, 14-22, 16-20, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23 or 24 carbon atoms.In some embodiments, for any one of Compounds (l)-(4), R55, for each occurrence, is independently selected from:-OC(=O)C(R42)[CH2]nC(=O)O-R41-CH(C4-Ci2alkyl)2, -R41-N(R42)([CH2]nC(=O)O-R41-CH(C4-Ci2 alkyl)2, -OC(=O)C(R43)[CH2]nC(=O)O-R41-CH(C4-Ci2 alkyl)2, -R41-N(R43)([CH2]nC(=O)O-R41- CH(C4-Ci2 alkyl)2, -R41-C(R42)[CH2]nC(=O)O-R41-CH(C4-Ci2alkyl)2or -R41-CH(R43)(R44), -R41- N(R43)(R44); wherein,R43for each occurrence, is independently selected from: -C(=O)O-C6-C24alkyl, -OC(=O)-C6-C24alkyl, -OC(=O)O-C6-C24alkyl, -C(=O)O-C6-C24alkenyl, -O-C(=O)-C6-C24alkenyl, -OC(=O)O-C6-C24alkenyl-, -C2-C22 alkylene-O-C2-C22 alkyl, -C2-C22 alkenylene-O-C2-C22 alkyl, -C2-C22 alkylene-O-C2-C22 alkenyl, -C1-C22 alkylene-C(=O)O-C1-C22 alkyl, -C1-C22 alkylene-O-C(=O)O-C1-C22 alkyl, - C1-C22 alkylene-O-C(=O)-C1-C22 alkyl, -C2-C22 alkenylene-C(=O)O-C1-C22 alkyl, -C2-C22 alkenylene-O-C(=O)-C1-C22 alkyl, -C2-C22 alkenylene-O-C(=O)-Ci-C22 alkyl, -C1-C22 alkylene- C(=O)O-C2-C22 alkenyl, -C1-C22 alkylene-O-C(=O)O-C2-C22 alkenyl, -C1-C22 alkylene-O-C(=0)-C2- C22 alkenyl, -C2-C22 alkenylene-C(=O)O-C2-C22 alkenyl, -C2-C22 alkenylene-O-C(=O)O-C2-C22 alkenyl, -C2-C22 alkenylene-O-C(=O)-C2-C22 alkenyl; wherein R43has 6-24 total carbon atoms;R44for each occurrence, is independently selected from: -R45-C(=O)O-R45-CH(C6-C12 a I ky 1)2, - R45- OC(=O) - R45-CH(C6-C12 alkyl)2, -R45-OC(=O)O-R45-CH(C6-C12 alkyl)2, R45-C(=O)O-R45-CH(C6- C12 alkenyl)2, -R45- OC(=O) - R45-CH(C6-C12 alkenyl)2, -R45-OC(=O)O- R45-CH(C6-C12 alkenyl)2; wherein R44has 6-24 total carbon atoms and wherein R45is, for each occurrence, independently selected from: a bond, C1-C6 alkylene or C1-C6 alkenylene; and each remaining variable, for each occurrence, is independently as described above for any one of Compounds (1) - (4) or for any embodiment of any compound herein.In some embodiments, for any one of Compounds (l)-(4), R55, for each occurrence, is independently selected from:wherein:R16for each occurrence, is independently selected from: a bond or — [CH2]k-; k is an integer selected from: 1, 2, 3, 4, 5 or 6;R62, R63, R67, R68, R74, R76, R108, and R110are each independently selected from: C1-C12 alkylene or C1-C12 alkenylene; R66, R71, R79, R106and R113are each independently selected from: C1-C12 alkyl or C1-C12 alkenyl;A3and A5are each independently selected from: -O-, -OC(=O)-, -C(=O)O- or -OC(=O)O-; andA4, A6, A7, A8, A12, A13, R64, R65, R69, R70, R75, R77, R78, R105, R109, R110, Rmand R112are a bond; and each remaining variable, for each occurrence, is independently as described above for any one of Compounds (1) - (4) or for any embodiment of any compound herein. In some further embodiments, R62, R63, R66and R74together, contain a total of 10-22 carbon atoms. In some other embodiments, R62, R63and R66, together, contain a total of 10-22 carbon atoms. In some other embodiments, R62, R63, R76and R79, together, contain a total of 10-22carbon atoms. In some other embodiments, R67, R68and R71, together, contain a total of ID- 22 carbon atoms. In some other embodiments, R67, R68, R108and R106, together, contain a total of 10-22 carbon atoms. In some other embodiments, R67, R68, R110and R113, together, contain a total of 10-22 carbon atoms. In some other embodiments, R63a bond. In other embodiments, R63and R68are absent.In some embodiments, for any one of Compounds ( l)-(4), R55, for each occurrence, is independently:wherein,A3and A5, for each occurrence, are each independently selected from: -O-, -OC(=O)-, - C(=O)O- or -OC(=O)O-;R16, for each occurrence, is independently selected from: a bond or -[CH2]k-; k is an integer selected from: 1, 2, 3, 4, 5 or 6; ccurrence, are each independently selected from:R62, R63, R67, R68, R74, R76, for each occurrence, are each independently selected from: C1-C12 alkylene or C1-C12 alkenylene;R66,R7ia nc|R79, for each occurrence, are each independently selected from: C1-C12 alkyl or C1-C12 alkenyl;R116, for each occurrence, and any remaining variable, for each occurrence, is independently as described above for any one of Compounds (1) - (4) or for any embodiment of any compound herein; and wherein, R62, R63, R66and R74, together, contain 10-22 total carbon atoms; R62, R63, R76and R79, together, contain 10-22 total carbon atoms; and R67, R68and R71, together, contain 10-22 total carbon atoms. In some other embodiments, R63a bond. In some other embodiments, R63and R68are absent.In some embodiments, -R62-A3-R63-R73-R74-R66, -R62-A3-R63-R73-R76-R79, and -R67-A5-R68-R71, independently has: c) a total number of carbon atoms selected from: 10-24, 12-22, 14-22, 16-20, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23 or 24 carbon atoms; d) a longest linear chain with a total number of carbon atoms that is selected from: 10- 24, 12-22, 14-22, 16-20, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23 or 24 carbon atoms; or e) a longest linear chain with a total number of atoms that is selected from: 10-24, 12-22, 14-22, 16-20, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23 or 24 atoms.In some embodiments, for any one of Compounds ( l)-(4), R55, for each occurrence, is independently:wherein:A3and A5, for each occurrence, are each independently selected from: -O-, -OC(=O)-, - C(=O)O- or -OC(=O)O-;R16for each occurrence, is independently selected from: a bond or -[CH2]k-; k is an integer selected from: 1, 2, 3, 4, 5 or 6;R61, R73and R107, for each occurrence, are each independently selected from:R62, R63, R67, R68, R74, R76, R108, and R110, for each occurrence, are each independently selected from: C1-C12 alkylene or C1-C12 alkenylene;R66, R79, R106and R113, for each occurrence, are each independently selected from: C1-C12 alkyl or C1-C12 alkenyl;R116and any remaining variables, for each occurrence, is independently as described above for any one of Compounds (1) - (4) or for any embodiment of any compound herein. In some further embodiments, R62, R63, R66and R74, together, contain 10-22 total carbon atoms; R62, R63, R76and R79, together, contain 10-22 total carbon atoms; and R67, R68, R108and R106, together, contain 10-22 total carbon atoms; and, R67, R68, R110and R113, together, contain 10-22 total carbon atoms. In some other embodiments, R63a bond. In other embodiments, R63and R68are absent. In some other embodiments, R73and R107areIn some embodiments, for any one of Compounds ( l)-(4), R55, for each occurrence, is independently:wherein:A3and A5, for each occurrence, are each independently selected from: -O-, -OC(=O)-, - C(=O)O- or -OC(=O)O-;R16, for each occurrence, is independently selected from: a bond or -[CH2]k-; k is an integer selected from: 1, 2, 3, 4, 5 or 6;R61, R73and R107, for each occurrence, are each independently selected from:R62, R63, R67, R68, for each occurrence, are each independently selected from: C1-C12 alkylene or C1-C12 alkenylene;R66and R71, for each occurrence, are each independently selected from: C1-C12 alkyl or C1-C12 alkenyl; R116and any remaining variables, for each occurrence, is independently as described above for any one of Compounds (1) - (4) or for any embodiment of any compound herein. In some further embodiments, R62, R63and R66together, contain 10-22 total carbon atoms. In some other embodiments, R67, R68and R71, together, contain 10-22 total carbon atoms. In some other embodiments, R63a bond. In other embodiments, R63and R68are a bond. In some embodiments, for any one of Compounds ( l)-(4), R55, for each occurrence, is independently selected from:; or; wherein,A3-A8, A12, A13, R62-R69, R73-R79, and R105-R113and any remaining variables, for each occurrence, are as defined above for any one of Compounds ( l)-(4) or for any embodiment of any compound herein. In some further embodiments, a compound selected from any one of Compounds (l)-(4) comprises a first and a second R55with longest linear chains that differ in length by 1, 2, 3, 4, 5, 6, 7, 8, 9 or 10 atoms. In some embodiments, said first and second R55have longest linear chains with a different number of carbon atoms. In some further embodiments, a first R55has a longest linear chain with 1, 2, 3, 4, 5, 6, 7, 8, 9 or 10 less carbon atoms than a longest linear chain of a second R55.In some embodiments, the ionizable cationic lipid has a structural formula selected from:Compound (5):Compound (7):(7), orCompound (8):(8); wherein:R16, for each occurrence, is independently selected from: a bond or — [CH2]k-;R62, R63, R64, R67, R68, R69, R74, R75, R76and R77, for each occurrence, are each independently selected from: a bond, C1-C24 alkylene or C1-C24 alkenylene; rence, are each independently selected from:R65, R70and R78, for each occurrence, are each independently selected from: a bond, - CH=CH-CH2-CH=CH- , -CH=CH-CH2-cPr-, -cPr-CH2-CH=CH-, -cPr-CH2-cPr-, -CH=CH-CD2-CH=CH- , -CH=CH-CD2-cPr-, -cPr-CD2-CH=CH- -or -cPr-CD2-cPr-, where -cPr- is cyclopropylene, each double bond or cPr has a cis configuration and D denotes deuterium (2H);R66, R71and R79, for each occurrence, are each independently selected from: C1-C24 alkyl or C1-C24 alkenyl;A3, A4, A5, A6, A7and A8and R1, for each occurrence, and each remaining variable, for each occurrence, is independently as described above for any one of Compounds ( l)-(4) or any embodiment of any compound herein. In some further embodiments, -R62-A3-R63-A4-R64-R65-R67-A5-R68-A6-R69-R70-R71, each independently has a total of 10-24 carbon atoms.In some embodiments, the ionizable cationic lipid is one of:Compound (9)(10),Compound (11)Compound (12)or a pharmaceutically acceptable salt or co-crystal thereof; wherein:R61. for each occurrence, is independently selected from:R85and R90, for each occurrence, are each independently selected from: a bond or -[CH2]I-8-;R86and R91, for each occurrence, are each independently selected from: a bond -O-, -O- C(O=)-, -O-C(=O)-O- or -C(=O)-O-;R87, for each occurrence, is independently selected from: a bond or -[CH2]I-6-;R88and R89, for each occurrence, are each independently -[CH2]I-IOCH3;R92, for each occurrence, is -[CH2]2-i6CH3;R93and R94, for each occurrence, are each independently selected from: -H, -D, -CH3, -CD3, - CH2CH3, -CD2CH3, where D denotes deuterium (2H);R95, R96, R100, R101and R102, for each occurrence, are each independently selected from: -OH, -O-C1-C4 alkyl, -O-C(=O)-O-Ci-C4alkyl or -O-C(=O)-Ci-C4alkyl;R97, for each occurrence, is independently selected from: -[CH2]I-6-, -[CH2]I-6-C(=O)-O-, -[CH2]I-6-C(=O)-O-, -[CH2]I-6-O- or -[CH2]I-6-C(=O)-, wherein:H V-when R61is Z or X , then R97is selected from: -[CH2]I-6- or -[CH2]I-6-C(=O)-O- and when R , then R97is selected from: -[CH2]I-6-, -[CH2]I-6-C(=O)-O-, -[CH2]I-6-O- or - [CH2]I-6R16, for each occurrence, is independently selected from: a bond or -[CH2]k-;R98, for each occurrence, is independently selected from: a bond, -C(=O)-, or -C(=O)-O-; andR104, for each occurrence, is independently selected from:wherein: R85-R92and R97, for each occurrence, are each independently as described above for any one of Compounds (8), (10) or (11); and wherein: each remaining variable, for each occurrence, is independently as described above for any one of Compounds (l)-(8) or any embodiment of any compound herein.In some embodiments, for any one of Compounds (1)-(12), R85and R90are the same. In some other embodiments, for any one of Compounds (1)-(12), R85and R90are each independently -[CH2]4-5--In some embodiments, for any one of Compounds (1)-(12), R86and R91are -C(O=)-O-. In some other embodiments, for any one of Compounds (1)-(12), R86and / or R91are / a bond. In some other embodiments, for any one of Compounds (1)-(12), R86and / or R91are / is -C(=O)- O-.In some embodiments, for any one of Compounds (1)-(12), R88and R89are each independently -[CH2]6-IO-.In some embodiments, for any one of Compounds (1)-(12), R92is -[CH2]8-I2-.In some embodiments, for any one of Compounds (1)-(12), R85, R86, R87, R88and R89, together, have between 14-24 or between 16-22 total carbon atoms.In some embodiments, for any one of Compounds (1)-(12), R90, R91and R92, together, have between 10-20 or between 12-18 total carbon atoms. In some embodiments, for any one of Compounds (1)-(12), R88and R89are identical.In some embodiments, for any one of Compounds (1)-(12), R85, R86, R87and R88, together have a total number of carbon atoms that differs by + / - 3, + / - 2 or + / -1, or is equal to the total number of carbon atoms of R90, R91and R92, combined.In some embodiments, for any one of Compounds (1)-(12),_R93and / or R94are / is -H. In some embodiments, _R95and / or R96are / is -OH.In some embodiments, for any one of Compounds (1)-(12), R85and R90are each independently -[CH2]4-5-; R86and R91are -C(O=)-O-; R88and R89are each independently - [CH2]6-IO-; R92is -[CH2]8-I2-; R93and / or R94are / is -H; and R95and / or R96are / is -OH.When R61is J- for any one of compounds 1-12, the carbon of R61may be a chiral center. In some embodiments, the chiral center is in an R configuration and in other embodiments, an S configuration.In some embodiments, the ionizable cationic lipid has a structural formula selected from any one of:Compound (13)(13),Compound (14)( ) or a pharmaceutically acceptable salt or co-crystal thereof; wherein:R80, for each occurrence, is independently selected from: a bond, R16or - [d-hJi-e-O- or -O-;R16, for each occurrence, is independently selected from: a bond or -[CFhJk-;R81, R82, R83, and R84, for each occurrence, are each independently selected from: a bond or - [CH2]I-IO-;R61, R93, R94, R95, R96, R97R98and each remaining variable, for each occurrence, is independently as described above for any one of Compounds ( l)-( 12) or any embodiment of any compound herein.In some embodiments, for any one of Compounds (1)-(19), R81and R83are each independently -[CH2]5-9-- In other embodiments, for any one of Compounds (1)-(19), R82and R84are each independently -[Cl-hjo-e-- In other embodiments, for any one of Compounds (1)- (19), R81and R83are each independently -[C]-hjs-io- and R82and R84are each independently - [CH2]O-6".In other embodiments, for any one of Compounds (1)-(19), R81and R82, combined, have between 7-13 total carbon atoms. In other embodiments, for any one of Compounds (1)- (19), R83and R84, combined, have between 7-13 total carbon atoms. In other embodiments for any one of Compounds ( l)-( 19), R81and R82, combined, have between 8-12 total carbon atoms. In other embodiments, for any one of Compounds (1)-(19), R83and R84, combined, have between 8-12 total carbon atoms. In other embodiments, for any one of Compounds (1)-(19), R81and R82, combined, and R83and R84, combined, each independently havebetween 7-13 or between 8-12 total carbon atoms. In other embodiments, for any one of Compounds (1)-(19), R81and R82, combined, and R83and R84, combined, each have identical number of total carbon atoms which is between 7-13 or between 8-12. In other embodiments, for any one of Compounds (1)-(19), R81and R82, combined, and R83and R84, combined, have a different number of total carbon atoms. In other embodiments, for any one of Compounds ( l)-( 19), when R81and R82, combined, and R83and R84, combined, have a different number of total carbon atoms, the carbon atom between R81and R83is a chiral center. In some further embodiments, the chiral center is in an R configuration and in other embodiments, an S configuration. In some embodiments, the ionizable cationic lipid has a structural formula selected from any one of:Compound (20)(22),Compound (23)(24),Compound (25)Compound (27)(28), or a pharmaceutically acceptable salt or co-crystal thereof; wherein: R61, R85, R86, R87, R88, R89, R90, R91, R92, R93, R94, R96, R97and each remaining variable, for each occurrence, is independently as described above for any one of Compounds (l)-( 19) or other embodiment of any compound herein.In some embodiments, the ionizable cationic lipid has a structural formula selected from:Compound (29)(29),Compound (30)(33),Compound (34)(35),Compound (36)(37),Compound (38)Compound (40)(41),Compound (42)Compound (46)Compound (48)(48) orCompound (49)(49),Compound (50R)Compound (53)or a pharmaceutically acceptable salt or co-crystal thereof; wherein:R1, R3, R16, R36, A1, and each remaining variable, for each occurrence, is independently as described above for any one of Compounds (l)-(28) or any embodiment of any compound herein.In some embodiments for an ionizable cationic lipid of the invention comprising a chiral center, e.g., as indicated by an *, the chiral center is in an R-configuration, in other embodiments an S-configuration, and in still further embodiments, either in an R- or an S- configuration.In some embodiments for an ionizable cationic lipid of the invention, R1and R3, for each occurrence, are each independently, -H, optionally substituted C1-C6 alkyl, optionally substituted C2-C6 alkenyl, optionally substituted C1-C6 hydroxyalkyl or optionally substituted C2-C6 hydroxyalkenyl. In other embodiments, R1and R3, for each occurrence, are each independently selected from: -H, methyl, ethyl, propyl, isopropyl, butyl, isobutyl or tertbutyl. In further embodiments, R2is absent or -H. In further embodiments, R1and R3, for each occurrence, are each independently selected from: -H, -D, -CH3, -CD3, -CH2CH3, - CD2CH3, where D denotes deuterium (2H); and R2is absent.In some embodiments for an ionizable cationic lipid of the present invention, R1and R3, for each occurrence, are each independently selected from: -H, C1-C6 alkyl, C2-C6 alkenyl, C1-C5 alkyl, C2-C5 alkenyl, C1-C4 alkyl, C2-C4 alkenyl, Ce alkyl, C5 alkyl, C4 alkyl, C3alkyl, C2 alkyl, Ci alkyl, C6alkenyl, or C5alkenyl, or C4alkenyl, or C3alkenyl, or C2alkenyl. In some furtherembodiments, R1and R3are each independently selected from: -H or C1-C3 alkyl. In some embodiments, RTand R3are identical.In some embodiments for an ionizable cationic lipid of the present invention, R2is absent.In other embodiments, for an ionizable cationic lipid of the present invention, R4, for each occurrence, is independently selected from:or ; wherein R41,R35, R55, R96and each remaining variable, for each occurrence, is independently as described above for any one of Compounds (l)-(54) or any embodiment of any compound herein.In some embodiments for an ionizable cationic lipid of the present invention, R6, for each occurrence, is independently selected from C3-C24 branched alkylene or C3-C24 branched alkenylene.In some embodiments for an ionizable cationic lipid of the present invention, R6, for each occurrence, is independently selected from: C1 -C3 alkylene, C1-C9 alkylene, C2-C9 alkenylene, C1-C7 alkylene, C2-C7 alkenylene, C1 -C5 alkylene, C2-C5 alkenylene, C2-C8 alkylene, C2-C8 alkenylene, C3-C7 alkylene, C3-C7 alkenylene, C5-C7 alkylene, C5-C7 alkenylene, C12 alkylene, CH alkylene, C10 alkylene, C9 alkylene, Cs alkylene, C7 alkylene, Ce alkylene, C5 alkylene, C4 alkylene, C3 alkylene, C2 alkylene, Ci alkylene, C12 alkenylene, C11 alkenylene, Cwalkenylene, C9alkenylene, C8 alkenylene, C7alkenylene, C6alkenylene, C5alkenylene, C4alkenylene, C3alkenylene, C2 alkenylene.In some embodiments for an ionizable cationic lipid of the present invention, R8and R9, for each occurrence, are each independently C6-C12 alkyl or C6-C12 alkenyl. In other embodiments, R8is Ce-Cio alkyl or Ce-Cio alkenyl. In other embodiments, R8and R9, together, have a total of 12-20 carbon atoms.In some embodiments for an ionizable cationic lipid of the present invention, R8, R9, R12, and R13, each independently have 1, 2 or 3, C=C double bonds. In further embodiments, at least one of the C=C double bonds is of Z configuration. In further embodiments, each C=C double bond is in a Z configuration. In further embodiments, for an ionizable cationic lipid of the invention, R14is C1-C3 alkyl, e.g., methyl or ethyl. In further embodiments, R8and R9, are each independently selected from: -C10-C18 alkenyl comprising 1, 2, 3 or 4 cis double bonds, - C10-C18 alkenyl ester comprising 1, 2, 3 or 4 cis double bonds, optionally substituted -C6-C18alkyl, -C6-718 alkyl ester, -C6-C18 alkyl ether, -C6-C18 alkyl carbonate, -C6-C18 alkenyl, -C6-C18 alkenyl ester, -C6-C18 alkenyl ether, or -C6-C18 alkenyl carbonate. In further embodiments, R8and R9, are each independently -C10-C18 alkenyl comprising 2 cis (Z configuration) double bonds. In some embodiments for an ionizable cationic lipid of the present invention, R8and R9are each independently selected from: optionally substituted C6-C18 alkyl, C6-718 alkylester, C6-C18 alkyl ether, C6-C18 alkyl carbonate C6-C18 alkenyl, C6-C18 alkenylester, C6-C18 alkenylether, or C6-C18 alkenylcarbonate.In some embodiments for an ionizable cationic lipid of the present invention, R8and R9, for each occurrence, are each independently selected from: C6-C14 alkyl, C6-C14 alkenyl, Cs-C12 alkyl, C8-C12 alkenyl, C16 alkyl, C15 alkyl, C14 alkyl, C13 alkyl, C12 alkyl, C11 alkyl, C10 alkyl, C9 alkyl, Cs alkyl, C7 alkyl, C16 alkenyl, C15 alkenyl, C14 alkenyl, C13 alkenyl, C12 alkenyl, C11 alkenyl, C10 alkenyl, C9, alkenyl, C8 alkenyl, or C7 alkenyl; wherein R8and R9, combined, have more than 15 total carbon atoms. In some embodiments, R8and R9have an equal number of carbon atoms or are identical: wherein R8and R9, combined, have more than 15 total carbon atoms. In some embodiments, R8and R9differ in total carbon atoms from each other; R8and R9differ by one or two total carbon atoms from each other; or R8and R9differ by one total carbon atoms.In some embodiments, for an ionizable cationic lipid of the present invention, R8is C7alkyl and R9is C8alkyl; R8is C8alkyl and R9is C7al kyl;R8is C8alkyl and R9is C9alkyl; R8is C9alkyl and R9is C8 alkyl; R8is C9 alkyl and R9is C10 a I kyl;R8is C10 alkyl and R9is C9 alkyl; R8is C10 alkyl and R9is C11 alkyl; R8is C11 alkyl and R9is C10 alkyl; R8is C11 alkyl and R9is C12 alkyl; R8is C12 alkyl and R9is C11 alkyl; R8is C7 alkyl and R9is C9 alkyl; R8is C9 alkyl and R9is C7 alkyl; R8is C8 alkyl and R9is C10 alkyl; R8is C10 alkyl and R9is C8 alkyl; R8is C9 alkyl and R9is C11 alkyl; R8is C11 alkyl and R9is C9alkyl; R8is C10 alkyl and R9is C12 alkyl; R8is C12 alkyl and R9is C10 alkyl; R8is C11 alkyl and R9is Cs alkyl; or R8is C8 alkyl and R9is C11 alkyl.In some embodiments for an ionizable cationic lipid of the present invention, R10and R11, for each occurrence, are each independently selected from: C1-C16 unbranched alkyl or C2-C16 unbranched alkenyl.In some embodiments, for an ionizable cationic lipid of the present invention, R16is C1-C3 alkylene.In some embodiments, for an ionizable cationic lipid of the present invention, R29, R30and R31are each -CH2-.In some embodiments, for an ionizable cationic lipid of the present invention, R32, R33and R34are each -O-.In some embodiments, for an ionizable cationic lipid of the present invention, R35is -OH.In some embodiments, for an ionizable cationic lipid of the present invention, R29, R30and R31are each -CH2-, and R32, R33and R34are each -O-.In some embodiments, for an ionizable cationic lipid of the present invention, R29, R30and R31are each -CH2-, R32and R33are each -O- and R35is -OH.In some embodiments for an ionizable cationic lipid of the present invention, R42is - [CH2]nC(=O)O[CH2]r; wherein n is as defined above for Compounds (l)-(4) or any embodiment of any compound herein; and r is an integer selected from 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16 or 17. In some embodiments for an ionizable cationic lipid comprising an R42variable, R42, for each occurrence, is independently selected from: -R46-C(=O)O-R47-, - R46-OC(=O)-R47-, -R46-OC(=O)O-R47-,or -R46-O-R47-, wherein; R46, for each occurrence, is independently selected from C1-C12 alkylene, C1-C12 alkylene ester, C1-C12 alkylene ether, Ci- Ci2alkylene carbonate; and R47, for each occurrence, is independently selected from -C1-C12 alkyl, -Ci-Ci2alkyl ester, -C1-C12 alkyl ether, or -C1-C12 alkyl carbonate.In some embodiments, for an ionizable cationic lipid of the present invention, R55for each occurrence, is independently:; wherein A1, R6, A2, R7, R8and R9, are as described for any one of Compounds (l)-(28) or any embodiment of any compound herein.In some embodiments, for an ionizable cationic lipid of the present invention, R55for each occurrence, is independently selected from:or wherein:R5for each occurrence is independently selected from: -H, optionally substituted C1-C16 alkyl, optionally substituted C1-C16 alkyl ester, optionally substituted C2-Ci6 alkenyl, optionally substituted C2-Ci6 alkenyl ester,; wherein, R10and R11are each independently selected from: C1-C12 unbranched alkyl, C2-C12, unbranched alkenyl, C2-Ci2unbranched alkyl, C2-C12unbranched alkenyl, C5-C7, unbranched alkyl, or C5-C7 unbranched alkenyl. In further embodiments, R10and R11are each independently selected from; C16 unbranched alkyl, C15 unbranched alkyl, C14 unbranched alkyl, C13 unbranched alkyl, C12 unbranched alkyl, C11 unbranched alkyl, C10 unbranched alkyl, C9 unbranched alkyl, Csunbranched alkyl, C7 unbranched alkyl, Ce unbranched alkyl, C5 unbranched alkyl, C4 unbranched alkyl, C3 unbranched alkyl, C2 unbranched alkyl, or Ci unbranched alkyl. In further embodiments, R10and R11are each independently selected from; C16unbranched alkenyl, C15 unbranched alkenyl, C14 unbranched alkenyl, C13 unbranched alkenyl, C12 unbranched alkenyl, C11 unbranched alkenyl, C10 unbranched alkenyl, C9 unbranched alkenyl, C8 unbranched alkenyl, C7 unbranched alkenyl, Ce unbranched alkenyl, C5 unbranched alkenyl, C4 unbranched alkenyl, C3 unbranched alkenyl, or C2 alkenyl; and R15, R6, A2, R7, R8, R9and any remaining variables are as described for any one of Compounds (l)-(54) or any embodiment of any compound herein. In further embodiments, R10and R11are each independently selected from; C2-C10 unbranched alkyl, or C2-C10 unbranched alkenyl. In further embodiments, R10and R11are identical.In some embodiments, for an ionizable cationic lipid of the present invention, R55for each occurrence, is independently selected from:wherein:R15, R5, R6, A2, R7, R8and R9is as described for any one of Compounds (l)-(54) or any embodiment of any compound herein.In some embodiments, for an ionizable cationic lipid of the present invention, R55for each occurrence, is independently selected from:or wherein:R15, R5, R6, R7, R8and R9is as described for any one of Compounds (l)-(54) or any embodiment of any compound herein.In some embodiments, for an ionizable cationic lipid of the present invention, R55for each occurrence, is independently selected from:or ; wherein: R15, R6, A2, R7, R8and R9is as described for any one of Compounds (l)-(54) or any embodiment of any compound herein.In some embodiments, for an ionizable cationic lipid of the present invention, R55for each occurrence, is independently selected from:A2, R7, R8and R9is as described for any one of Compounds (l)-(54) or any embodiment of any compound herein.In some embodiments for an ionizable cationic lipid of the present invention, R5for each occurrence is independently selected from: C1-C14 unbranched alkyl, C2-C14 unbranched alkenyl, or, wherein: R10and R11for each occurrence, are each independently selected from: C1-C12 unbranched alkyl, C2-C12, unbranched alkenyl, C2-C12 unbranched alkyl, C2-C12 unbranched alkenyl, C5-C7, unbranched alkyl, or C5-C7 unbranched alkenyl. In further embodiments, R10and R11are each independently selected from; Ci6 unbranched alkyl, C15 unbranched alkyl, C14 unbranched alkyl, C13 unbranched alkyl, C12 unbranched alkyl, Cn unbranched alkyl, C10 unbranched alkyl, C9 unbranched alkyl, Cs unbranched alkyl, C7 unbranched alkyl, Ce unbranched alkyl, C5 unbranched alkyl, C4 unbranched alkyl, C3 unbranched alkyl, C2 unbranched alkyl, or Ci unbranched alkyl. In further embodiments, R10and R11are each independently selected from; Ci6 unbranched alkenyl, C15 unbranched alkenyl, C14 unbranched alkenyl, C13 unbranched alkenyl, C12 unbranched alkenyl, Cn unbranched alkenyl, C10 unbranched alkenyl, C9 unbranched alkenyl, Cs unbranched alkenyl, C7 unbranched alkenyl, Ce unbranched alkenyl, C5 unbranched alkenyl, C4 unbranched alkenyl, C3 unbranched alkenyl, or C2 alkenyl. In further embodiments, R10and R11are each independently selected from; C2-C10 unbranched alkyl, or C2-C10 unbranched alkenyl. In further embodiments, R10and R11are identical. In some embodiments, R5, for each occurrence, is independently selected from: C1-C16 unbranched alkyl, C1-C16 unbranched alkyl ester, unbranched C2-C16 alkenyl or C2-C16 unbranched alkenyl ester. In some embodiments, R5, for each occurrence, is independently selected from:C3-Ci6 branched alkyl, C3-C16 branched alkyl ester, C3-C16 branched alkenyl, or C3-C16 branched alkenyl ester. In some embodiments, for an ionizable cationic lipid of the present invention, R55for each occurrence, is independently selected from: -OC(=O)C(R42)[CH2]nC(=O)O-R41-CH(C4-Ci2 alkyl)2, -R41-N(R42)([CH2]nC(=O)O-R41-CH(C4-Ci2 alkyl)2, -OC(=O)C(R43)[CH2]nC(=O)O-R41-CH(C4- C12 alkyl)2, -R41-N(R43)([CH2]nC(=O)O-R41-CH(C4-Ci2 alkyl)2, -R41-C(R42)[CH2]nC(=O)O-R41- CH(C4-Ci2 alkyl)2, -R41-CH(R43)(R44), or -R41-N(R43)(R44); whereinR41-R45and each remaining variable, for each occurrence, is independently as described above for any one of Compounds (l)-(54) or any embodiment of any compound herein.In some embodiments for an ionizable cationic lipid of the present invention, A2is -OC(=O)-, -OC(=O)R16C(=O)O- or -C(=O)O-.In some embodiments for an ionizable cationic lipid of the present invention, each R55has 12-50, 18-30, 20-30, or 12-24 total carbon atoms. In further embodiments, each R55comprises 0-6, 0-3, 1-3, 1, 2, 3, 4, 5 or 6 esters; 0-6, 0-3, 1-3, 1, 2, 3, 4, 5 or 6 ethers; and / or 0-6, 0-3, 1-3, 1, 2, 3, 4, 5 or 6 carbonates.In some embodiments for an ionizable cationic lipid of the present invention, R55for each occurrence, is independently selected from:-O-C(=O)C(R42)[CH2]nC(=O)O-R41-CH(C4-Ci2 alkyl)2, -R41-N(-R42)(-[CH2]nC(=O)O-R41-CH(C4-Ci2alkyl)2, -O-C(=O)C(R43)-[CH2]nC(=O)O-R41-CH(C4-Ci2alkyl)2, -R41-N(-R43)(-[CH2]nC(=O)O-R41- CH(C4-CI2alkyl)2, -R41-C(R42)[CH2]nC(=O)O-R41-CH(C4-Ci2alkyl)2, -R41-CH(R43)(R44), or -R41- N(R43)(R44).In some embodiments for an ionizable cationic lipid of the present invention, R55for each occurrence, is independently selected from: -R16-N(-R48)-R49, -R16-CH(-R48)-R49, -R16-OC(=O)- R16-N(-R48)-R49, -R16-OC(=O)CH(-R48)-R49, or -R16-C(=O)O-R16-CH(-R48)-R49; wherein: a) R48is -R16-O-CH(-[CH2]5-9 CH3)2and R49is -R16-C(=O)O-[CH2]6-I2CH3, b) R48is -CH2O-C(=O)O-CH(-[CH2]7CH3)2and R49is -R16-O-C(=O)O-[CH2]7-I3CH3, c) R48is -R16C(=O)O-CH([CH2]7CH3)2and R49is -R16C(=O)O-[CH2]7CH3, d) R48is -[CH2]3C(=O)O-CH(-[CH2]7CH3)2and R49is -[CH2]3C(=O)O-[CH2]7CH3, e) R48is -CH2C(=O)O-CH(-[CH2]9CH3)2and R49is -C(=O)O-[CH2]I0CH3, f) R48is -[CH2]3C(=O)O-CH(-[CH2]7CH3)2and R49is -[CH2]3C(=O)O-[CH2]9CH3 g) R48is -[CH2]2C(=O)O-CH(-[CH2]7CH3)2and R49is -[CH2]2C(=O)O-[CH2]9CH3, or h) R48is -R16-C(=O)O-R16-CH(-R57-CH3)2and R49is -[CH2]8-I6CH3;R16, for each occurrence, is independently selected from: a bond or -[CH2]k-;R57, for each occurrence, is independently selected from: a bond or -[CH2]m-; wherein m is an integer selected from: 1, 2, 3, 4, 5, 6, 7 or 8; and each remaining variable, for each occurrence, is independently as described above for any Compound or for any other embodiment of any compound herein.In some embodiments, for an ionizable cationic lipid of the present invention, R55for each occurrence, is independently selected from:; wherein: R16, R57, A1, A2, R61, R85, R86, R87, R88, R89, R90, R91, R92, R97and each remaining variable, for each occurrence, is independently as described above for any one of Compounds (l)-(54) or any embodiment of any compound herein.In some embodiments, R55for each occurrence, is independently selected from:; wherein:R16, R57, A1, R61, R81, R82, R83, R84, R97and each remaining variable, for each occurrence, is independently as described above for any one of Compounds (l)-(54) or any embodiment of any compound herein; and wherein each cyclopropyl (cPr) fused to adjacent chain4 carbons, i.e., , and each -C=C-, as shown in any one of the structures, has a cis (orE) configuration.In some embodiments, R55for each occurrence, is independently selected from:or; wherein:R16, R57, A1, and each remaining variable, for each occurrence, is independently as described above for any one of Compounds (l)-(54) or any embodiment of any compound herein; and wherein each -C=C-, as shown in any one of the structures has a cis configuration. In some embodiments, R55for each occurrence, is independently selected from:; wherein, R16, R61, R62 and R67, for each occurrence, are each independently as described above for any one of Compounds (1)- (54) or any embodiment of any compound herein; and wherein each -C=C-, as shown in any one of the structures has a cis configuration. In some embodiments, R55is:In one aspect, the ionizable cationic lipid has a structural formula selected from the group consisting of: Compound (121):(121),Compound (122):(122),Compound (123):(123),Compound (124):(124),Compound (125):(126),Compound (127):(127), Compound (128):Me is methyl; and,R55is as defined for any one of Compounds (l)-(54) or for any embodiment herein.In another aspect, the present invention provides for a lipid comprising lipid tail, wherein the tail is one more -R55. In some embodiments, the lipid further comprises a head group and an optional linker. In some embodiments, the head group is an amine containing group. In some further embodiments, the head group is an amine containing ionizable cationic head group.In some embodiments the ionizable cationic lipid is a non-fusogenic lipid. By a "non- fusogenic lipid" is meant a cationic lipid that can condense and / or encapsulate a payload, e.g., a therapeutic nucleic acid, but has insufficient fusogenic activity to effectively delivery the payload across cellular membranes. In some embodiments, the lipid nanoparticles have mean diameter of 20-75 nm or 30-100 nm.The pKa', i.e., the apparent pKa, of formulated cationic lipids in particles, can be correlated with the effectiveness of the LNPs for delivery of nucleic acids. The pKa' of a cationic lipid can be determined in lipid nanoparticles, e.g., using an assay based on fluorescence of 2-(p-toluidino)-6-napthalene sulfonic acid (TNS). Lipid nanoparticles in PBS at a concentration of 0.4 mM total lipid are prepared using standard methods. TNS can be prepared as a 100 mM stock solution in distilled water and mixed into buffers of different pH. Vesicles can be diluted to 24 mM lipid in 2 mL of buffered solutions containing, 10 mM HEPES, 10 mM MES, 10 mM ammonium acetate, 130 mM NaCI, where the pH ranges from 2.5 to 11. An aliquot of the TNS solution can be added to give a final concentration of 1 mM and following vortex mixing fluorescence intensity is measured at room temperature in a SLM Aminco Series 2 Luminescence Spectrophotometer using excitation and emission wavelengths of 321 nm and 445 nm. A sigmoidal best fit analysis can be applied to the fluorescence data and the pKa' is measured as the pH giving rise to half-maximal fluorescence intensity.In one embodiment, as a molar percent of total lipids, the LNP comprises about 30% to about 80%, about 30% to about 70%, about 30% to about 60%, about 30% to about 50%, or about 30% to about 40%, about 40% to about 80%, about 30% to about 70%, about 40% to about 60%, about 40% to about 50%, about 50% to about 80%, about 50% to about 70%, 50% to about 60%, about 60% to about 80%, or about 70% to about 80% (ionizable) cationic lipid.SterolIn one embodiment, the lipid particles (e.g., lipid nanoparticles) can further comprise a component, such as a sterol, to provide membrane integrity and stability of the lipid particle. In one embodiment, an exemplary sterol that can be used in the lipid particle is cholesterol, or a derivative thereof. Non-limiting examples of cholesterol derivatives include 5-a-cholestanol (5a-Cholestan-3|3-ol), 5-|3-coprostanol, cholesteryl-(2'-hydroxy)-ethyl ether, cholesteryl-( 4' -hydroxy)-butyl ether, and 6-ketocholestanol, 5a-cholestane, cholestenone, 5-a-cholestanone, 5 p-cholestanone, allocholesterol, epi-allocholesterol and cholesteryl decanoate; and mixtures thereof. In some embodiments, the cholesterol derivative is a polar analogue such as cholesteryl-(4'-hydroxy)-butyl ether. In some embodiments, cholesterol derivative is cholesteryl hemisuccinate. In some embodiments, the sterol is a sea cucumber sulphated sterol, e.g., cholest-5-en-3|3-yl hydrogen sulfate or 24-methylene- cholesterol sulfate (J. Oleo Sci. 71, (3) 401-410 (2022)). In some embodiments, the lipid is fecosterol, sitosterol, campesterol, stigmasterol, brassicasterol, ergosterol, tomatidine, tomatine, ursolic acid or alpha-tocopherol.Exemplary cholesterol derivatives are described in International Patent Application Publication No. W02009 / 127060 and U.S. Patent Application Publication No. US2010 / 0130588.In one embodiment, the component providing membrane integrity, such as a sterol, can comprise 0-50% (mol) of total lipids present in the lipid particle (e.g., lipid nanoparticle). In some embodiments, such a component is 20-50% (mol) of total lipids present in the lipid particle (e.g., lipid nanoparticle). In some embodiments, such a component is 30-40% (mol) of total lipids present in the lipid particle (e.g., lipid nanoparticle). In some embodiments,such a component is 35-45% (mol) of total lipids present in the lipid particle (e.g., lipid nanoparticle). In some embodiments, such a component is 38-42% (mol) of total lipid present in the lipid particle (e.g., lipid nanoparticle).According to some embodiments, the LNP comprises a sterol, wherein the sterol is present at a molar percentage of about 20% to about 50%. According to some embodiments, the LNP comprises a sterol, wherein the sterol is present at a molar percentage of about 30% to about 50%. According to some embodiments, the LNP comprises a sterol, wherein the sterol is present at a molar percentage of about 40% to about 50%. According to some embodiments, the LNP comprises a sterol, wherein the sterol is present at a molar percentage of about 20% to about 40%. According to some embodiments, the LNP comprises a sterol, wherein the sterol is present at a molar percentage of about 30% to about 40%. In some embodiments, the LNP comprises more than one structural lipid, e.g., two or more sterols.Non-cationic lipidThe non-cationic lipid is typically a phospholipid and serves to increase fusogenicity and / or increase stability of the LNP, including during formation. Non-cationic lipids include amphipathic lipids, neutral lipids and anionic lipids. Accordingly, the non-cationic lipid can be a neutral uncharged, zwitterionic, or anionic lipid. Exemplary non-cationic lipids include, but are not limited to, distearoyl-sn-glycerophosphoethanolamine, distearoylphosphatidylcholine (DSPC), dioleylphosphatidylcholine (DOPC), dipalmitoylphosphatidylcholine (DPPC), dioleylphosphatidylglycerol (DOPG), dipalmitoylphosphatidylglycerol (DPPG), dioleyl-phosphatidylethanolamine (DOPE), palmitoyloleylphosphatidylcholine (POPC), palmitoyloleylphosphatidylethanolamine (POPE), dioleyl-phosphatidylethanolamine 4-(N-maleimidomethyl)-cyclohexane-l-carboxylate (DOPE-mal), dipalmitoyl-phosphatidylethanolamine (DPPE), dimyristoylphosphoethanolamine (DMPE), distearoyl-phosphatidylethanolamine (DSPE), monomethylphosphatidylethanolamine (such as 16-O-monomethyl PE), dimethylphosphatidylethanolamine (such as 16-O-dimethyl PE), 18-1-trans PE, l-stearoyl-2-oleyl- phosphatidyethanolamine (SOPE), hydrogenated soy phosphatidylcholine (HSPC), egg phosphatidylcholine (EPC), dioleyl-phosphatidylserine (DOPS), sphingomyelin (SM), 1,2- diundecanoyl-sn-glycero-phosphocholine (DUPC), dimyristoyl-phosphatidylcholine (DMPC), dimyristoyl-phosphatidylglycerol (DMPG), distearoyl-phosphatidylglycerol (DSPG), dierucoyl- phosphatidylcholine (DEPC), palmitoyl-oleyl-phosphatidylglycerol (POPG), dielaidoylphosphatidylethanolamine (DEPE), l,2-dilauroyl-sn-glycero-3-phosphoethanolamine (D LPE); l,2-diphytanoyl-sn-glycero-3-phosphoethanolamine (DPHyPE); 1,2-di-O-octadecenyl- sn-glycero-3-phosphocholine (18:0 Diether PC), l-oleyl-2-cholesterylhemisuccinoyl-sn- glycero-3-phosphocholine (OChemsPC), l-hexadecyl-sn-glycero-3-phosphocholine (C16 Lyso PC), l,2-dilinolenoyl-sn-glycero-3- phosphocholine, 1 ,2-diarachidonoyl-sn-glycero-3- phosphocholine, 1 ,2-didocosahexaenoyl-sn-glycero-3-phosphocholine, 1,2-diphytanoyl-sn-glycero-3-phosphoethanolamine (ME 16.0 PE), l,2-distearoyl-sn-glycero-3- phosphoethanolamine, 1.2-dilinoleyl-sn-glycero-3-phosphoethanolamine, 1,2-dilinolenoyl- sn-glycero-3-phosphoethanolamine, ,2-diarachidonoyl-sn-glycero-3-phosphoethanolamine, l,2-didocosahexaenoyl-sn-glycero-3-phosphoethanolamine, phosphatidylethanolamine, lysolecithin, lysophosphatidylethanolamine, phosphatidylserine, phosphatidylinositol, sphingomyelin (SM), egg sphingomyelin (ESM), dihydrosphingomyelin, cephalin, cardiolipin, phosphatidicacid, cerebrosides, dicetylphosphate, lysophosphatidylcholine, dilinoleyl- phosphatidylcholine, or mixtures thereof. It is to be understood that other diacylphosphatidylcholine and diacylphosphatidylethanolamine phospholipids can also be used. The acyl groups in these lipids are preferably acyl groups derived from fatty acids having C10-C24 carbon chains, e.g., lauroyl, myristoyl, palmitoyl, stearoyl, or oleyl. Preferred helper lipid: DSPC, DPPC, DMPC, DOPC, POPC, DOPE, and SM.In some embodiments, the non-cationic lipid can comprise 0-20% (mol) of the total lipid present in the lipid nanoparticle. In some embodiments, the non-cationic lipid comprises 0.5-15% (mol) of the total lipid present in the lipid particle (e.g., lipid nanoparticle). In some embodiments, the non-cationic lipid comprises 5-12% (mol) or 5-10% (mol) of the total lipid present in the lipid particle (e.g., lipid nanoparticle). In some embodiments, the non-cationic lipid comprises about 6% (mol), about 7.0% (mol), about 7.5% (mol), about 8.0% (mol), about 9.0% (mol), about 10% (mol), or about 11 % (mol) of the total lipid present in the lipid particle (e.g., lipid nanoparticle).Exemplary non-cationic lipids are described in International Patent Application Publication No. WO2017 / 099823 and US Patent Application Publication No. US2018 / 0028664, the contents of both of which are incorporated herein by reference in their entirety.According to some embodiments, the LNP comprises a non-cationic lipid, wherein the noncationic lipid is present at a molar percentage of about 2 % to about 20%. According to some embodiments, the LNP comprises a non-cationic lipid, wherein the non-cationic lipid is present at a molar percentage of about 5% to about 20%, about 10% to about 20%, about 15% to about 20%, about 10% to about 20%, or about 10% to about 15%.Polymer conjugated lipidsIn one embodiment, the lipid particle (e.g., lipid nanoparticle) can further comprise a polymer conjugated lipid molecule, e.g., polyethylene glycol (PEG)-conjugated ("PEGylated") lipid. Generally, these are used to inhibit aggregation of lipid particle (e.g., lipid nanoparticle) and / or provide steric stabilization. Exemplary conjugated lipids include, but are not limited to, PEG-lipid conjugates, polyoxazoline (POZ)-lipid conjugates, polyamide - lipid conjugates (such as ATTA-lipid conjugates), cationic-polymer lipid (CPL) conjugates, and mixtures thereof. In some embodiments, the conjugated lipid molecule is a PEGylated lipid, for example, a (methoxy polyethylene glycol)-conjugated lipid. In some other embodiments, the PEGylated lipid is PEG2000-DMG (dimyristoylglycerol). Exemplary PEGylated lipidsinclude, but are not limited to, PEG-diacylglycerol (DAG) (such as l-(monomethoxy- polyethyleneglycol)-2,3-dimyristoylglycerol (PEG-DMG)), PEG-dialkyloxypropyl (DAA), PEG- phospholipid, PEG-ceramide (Cer), a pegylated phosphatidylethanolamine (PEG-PE), PEG succinate diacylglycerol (PEGS-DAG) l,2-dimyristoyl-rac-glycero-3-methoxypolyethylene glycol-2000 (DMG-PEG 2000), R-3-[((jo-methoxy polyethylene glycol)2000)carbamoyl)]-l,2- dimyristyloxy-propyl-3-amine (PEG-c-DOMG), l,2-Dimyristoyl-sn-Glycero-3-Phosphoethanolamine (DMPE) conjugated Polyethylene Glycol, l,2-Dilauroyl-sn-Glycero-3- Phosphoethanolamine (DLPE) conjugated Polyethylene Glycol, dipalmitoyl phosphatidylcholine (DPPC) conjugated Polyethylene Glycol, 1, 2-Distearoyl-sn-glycero-3- phosphoethanolamine-Poly(ethylene glycol) (DSPE-PEG), PEG dialkoxypropylcarbamate, N- (carbonyl-methoxypolyethylene glycol 2000)-l,2-distearoyl-sn-glycero-3- phosphoethanolamine sodium salt,or a mixture thereof. PEG mono-fatty acid esters may have hydroxy on the PEG terminus opposite to the hydrophobic lipid end, as inA polymer conjugated lipid may be used in a LNP formulation, alone or in mixtures with other surface lipids.Additional exemplary PEG-lipid conjugates are described, for example, in US patents US5885613, US6287591, US8936942B2 and US patent applications US2003 / 0077829, US2003 / 0077829, US2005 / 0175682, US2008 / 0020058, US2011 / 0117125, US2010 / 0130588, US2016 / 0376224, and US2017 / 0119904, the contents of all of which are incorporated herein by reference in their entireties.In some embodiments, the PEG-DAA PEGylated lipid is, for example, PEG- dilauroyloxypropyl, PEG- dimyristyoloxypropyl, PEG-dipalmitoyloxypropyl, or PEG- distearoyloxypropyl. The PEG-lipid can be, e.g., one or more of PEG-DMG, PEG- dilauroylglycerol, PEG-dipalmitoylglycerol, PEG-disteroylglycerol, PEG-dilauroylglycamide, PEG-dimyristoylglycamide, PEG-dipalmitoylglycamide, PEG- disteroylglycamide, PEG- cholesterol (l-[8'-( Cholest-5-en-3 [beta]-oxy)carboxamido-3',6'-dioxaoctanyl] carbamoyl- [omega]-methyl-poly(ethylene glycol), PEG-DMB (3,4-Ditetradecoxylbenzyl-[omega]-methyl- poly(ethylene glycol) ether), or 1,2-dimyristoyl-sn-glycero- 3-phosphoethanolamine-N- [methoxy(polyethylene glycol)-2000] . In some embodiments, the PEG-lipid is selected from PEG-DMG and / or l,2-dimyristoyl-sn-glycero-3- phosphoethanolamine-N- [methoxy(polyethylene glycol)-2000].In some embodiments, the PEGylated lipid is selected from one or more of: N-(Carbonyl- methoxypolyethyleneglycoln)-l,2-dimyristoyl-sn-glycero-3 -phosphoethanolamine (DMPE- PEGn, where n (representing PEG average molecular weight) is 350, 500, 750, 1000 or 2000); N-(Carbonyl-methoxypolyethyleneglycoln)-l,2-distearoyl-sn-glycero-3- phosphoethanolamine (DSPE-PEGn, where n is 350, 500, 750, 1000, 2000, or 5000); DSPE- polyglycerin-cyclohexyl-carboxylic acid, DSPE-polyglycelin-2-methylglutaric acid; 1,2- Distearoyl-sn-Glycero-3-Phosphoethanolamine (DSPE) conjugated Polyethylene Glycol (DSPE-PEG-OH); polyethylene glycol-dimyristolglycerol (PEG-DMG); polyethylene glycoldistearoyl glycerol (PEG-DSG); or N-octanoyl-sphingosine-l-{succinyl[methoxy(polyethylene glycol)2000 (Cs PEG2000 Ceramide). In some examples of DMPE-PEGn, where n is 350, 500, 750, 1000 or 2000, the PEG-lipid is N-(Carbonyl-methoxypolyethyleneglycol 2000)-l,2- dimyristoyl-sn-glycero-3-phosphoethanolamine (DMPE-PEG 2,000). In some examples of DSPE-PEGn, where n (representing PEG average molecular weight) is 350, 500, 750, 1000 2000, or 5000 the PEG-lipid is N-(Carbonylmethoxypolyethyleneglycol 2000)-l,2-distearoyl- sn-glycero-3-phosphoethanolamine (DSPE-PEG 2000). In some embodiments, the PEGylated lipid is DSPE-PEG-OH. In some embodiments, the PEGylated lipid is DSPE-PEG-azide. In some embodiments, the PEGylated lipid is PEG-DMG. In some embodiments, the PEGylated lipid is PEG-DSG. In some embodiments, the conjugated lipid, e.g., PEGylated lipid, includes a tissue-specific ligand, e.g., first or second ligand. For example, DSPE-PEG conjugated with a GalNAc ligand, DSGPEG conjugated with a GalNAc ligand. In one embodiment, lipids conjugated with a molecule other than a PEG can also be used in place of PEG-lipid. For example, polyoxazoline (POZ)-lipid conjugates, polyamide-lipid conjugates, such as ATTA- lipid conjugates and polysarcosine (PSAR) lipid conjugates, and cationic polymer lipid (CPL) conjugates, such as with chitosan free amine oligomers, can be used in place of or in addition to the PEG-lipid. Exemplary conjugated lipids, i.e., PEG-lipids, POZ-lipid conjugates, ATTA-lipid conjugates and cationic polymer-lipids are described in the International Patent Application Publication Nos. WO 1996 / 010392, WO1998 / 051278, W02002 / 087541, W02005 / 026372, WO2008 / 147438, W02009 / 086558, W02012 / 000104, WO2017 / 117528, WO2017 / 099823, WO2015 / 199952, W02017 / 004143, WO2015 / 095346, W02012 / 000104, W02012 / 000104, and W02010 / 006282, U.S. Patent Application Publication Nos.US2003 / 0077829, US2005 / 0175682, US2008 / 0020058, US2011 / 0117125, US2013 / 0303587, US2018 / 0028664, US2015 / 0376115, US2016 / 0376224, US2016 / 0317458, US2013 / 0303587, US2013 / 0303587, and US20110123453, and U.S. Patent Nos. US5,885,613, US6,287,591, US6, 320,017, and US6,586,559.According to some embodiments, the LNP comprises at least one PEGylated lipid, wherein the PEGylated lipid is present at a molar percentage of about 0.5% to about 20% of the total lipid present in the lipid nanoparticle. In some embodiments, the LNP comprises at least one PEGylated lipid, wherein the PEGylated lipid is present at a molar percentage of about 2.1% to about 10% of the total lipid present in the lipid nanoparticle. In some embodiments, the LNP comprises about 1-5% (mol), about 2-4% (mol), about 2-3% (mol), about 1-3% (mol),about 0.75-2.5% (mol), about 0.75-2.0% (mol), about 0.75-1.8% (mol), about 1-2% (mol), about 0.75-1.5% (mol), about 1- 1.8% (mol), about 1-1.5% (mol), about 1-1.3% (mol), about 1-1.2% (mol), about 0.75-1.5% (mol), about 0.75-1.25% (mol), about 1.5-1.8% (mol), about 1.2-1.5% (mol), about 2% (mol), about 2.5% (mol), about 3% (mol), about 3.5% (mol) or about 4% (mol) of PEGylated lipid, based on the total lipid present in the lipid nanoparticle. In some embodiments, the LNP comprises at least one PEGylated lipid, wherein the PEGylated lipid is present at a molar percentage of about 5% to about 10%, about 7% to about 10%, about 2.1 % to about 8%, about 2.1 % to about 5%, about 5% to about 8%, about 1 % to about 2%. about 1.2% to about 2%. about 1.5% to about 2%. about 1.75% to about 2%, about 1 % to about 1.5%, about 1.25% to about 1.5%, or about 1.5% to about 1.75%.In some embodiments, the lipid particles (e.g., lipid nanoparticles) is conjugated with other moieties to prevent aggregation. Such lipid conjugates include, but are not limited to, PEG- lipid conjugates such as, e.g., PEG coupled to dialkyloxypropyls (e.g., PEG-DAA conjugates), PEG coupled to diacylglycerols (e.g., PEG-DAG conjugates), PEG coupled to cholesterol, PEG coupled to phosphatidylethanolamines, and PEG conjugated to ceramides (see, e.g., U.S. Patent No. 5,885,613), cationic PEG lipids, polyoxazoline (POZ)-lipid conjugates (e.g., POZ- DAA conjugates; see, e.g., U.S. Provisional Application No. 61 / 294,828, filed Jan. 13, 2010, and U.S. Provisional Application No. 61 / 295,140, filed Jan. 14, 2010), polyamide oligomers (e.g., ATTA-lipid conjugates), and / or mixtures thereof. Additional examples of POZ-lipid conjugates are described in PCT Publication No. WO 2010 / 006282. PEG, PSAR or POZ can be conjugated directly to the lipid or may be linked to the lipid via a linker moiety. Any linker moiety suitable for coupling the PEG or the POZ to a lipid can be used including, e.g., nonester containing linker moieties and ester-containing linker moieties. In certain preferred embodiments, non-ester containing linker moieties, such as amides or carbamates, are used. The disclosures of each of the above patent documents are herein incorporated by reference in their entirety for all purposes.Additional LipidsLipid nanoparticles, including those of the present invention, may comprise one or more additional lipid(s), including triglycerides, fatty acids, carotenoids (including carotenes and xanthophylls), other classes of lipids, and derivatives (including conjugates) thereof. Note that some of these additional lipids may also fall into one of the categories above.Additional lipids may be present to improve stability, fusogenicity, endosomal escape, tolerability / safety, efficacy, tissue tropism / targeting, etc.One aspect of the invention provides for a lipid particle, e.g. LNP, comprising an additional lipid. These lipid particles include but are not limited to lipid particles comprising an ionizable cationic lipid of the present invention.In some embodiments, the lipid particle comprises a lipid selected from:Compound (55A)orCompound (55B)(55B).In another embodiment, the lipid particle comprises a carotene or xanthophyll. In a further embodiment, the lipid particle comprises a xanthophyll selected from astaxanthin, zeaxanthin, lutein, cryptoxanthin, antheraxanthin, fucoxanthin, neoxanthin or violaxanthin.The chemical structure:is free astaxanthin with the S,S'-stereochemistry of the secondary alcohol carbons.The xanthophyll may be esterified or unesterified, present as a single isomer, a racemate or mixture of isomers. Double bonds may be in an E or Z configuration. Astaxanthin, e.g., can exist as a (3S, 3'S), (3S, 3'R) or (3R, 3'R) isomer. In another embodiment, said compound is selected from any one of Compounds (56) - (70):: wherein A9, A10 and All are each independently selected from: R103or -H; wherein at least one of A9, A10 or All is R103; and wherein R103, for each occurrence, is independently selected from:In another embodiment, said compound is selected from any one of Compounds (71)-(120):10 (81),(82),(83),(84),(86),(87),(88),(89),(91),(92),10 (102),10 (107),(119), or(120).Therapeutic Nucleic Acids (TNA)The lipid particles of the present invention comprise a payload / cargo. In one aspect the payload is therapeutic nucleic acid (TNA). The length of the TNA can vary and include nucleic acid of 5-50,000 nucleotides in length. The nucleic acid can be in any form, including singlestranded DNA, single-stranded RNA, double-stranded DNA or double-stranded RNA, or hybrids thereof. The nucleotides may be modified, unmodified or a combination thereof. The TNA may be chemically synthesized. Synthesis of mRNA includes in vitro transcription. In some embodiments, the TNA is selected from the group consisting of minigenes, plasmids, minicircles, antisense oligonucleotides (ASO), closed-ended (ceDNA), ministring, doggybone, protelomere closed ended DNA, dumbbell linear DNA, asymmetrical interfering RNA (aiRNA), microRNA (miRNA), messenger RNA (mRNA), small interfering RNA (siRNA), small activating RNA (saRNA), self-amplifying RNA (SAM), ribozymes, dicer substrate dsRNA, small hairpin RNA (shRNA), tRNA, tRNA derived RNA fragments (tRFs), rRNA, piwi-interacting RNA (piRNA), guide RNA (gRNA), DNA viral vectors, viral RNA vector, non-viral vector and any combination thereof.The payload of an LNP may include more than one TNA. For example, in the case of Cas9- CRISPR, a guide RNA (gRNA), together with a plasmid or mRNA encoding the Cas9 protein may be incorporated into a single LNP. Alternatively, LNPs containing different TNAs may be combined into a single formulation.According to some embodiments, the LNP has a total lipid to TNA ratio of about 10: 1 to about 40:1, for example 10:1 to 30:1 or 10:1 to 20:1 or 10:1 to 15:1.A preferred payload of an LNP of the present invention is an mRNA. An mRNA of the invention may include a nucleic acid sequence encoding a polypeptide of interest (e.g., a coding region), a first flanking region located at the 5'-terminus of the first region (e.g., a 5'- UTR), a second flanking region located at the 3'-terminus of the first region (e.g., a 3'-UTR), at least one 5'-cap region, and a 3 '-stabilizing region. In some embodiments, a messenger RNA further includes a poly-A region or a Kozak sequence (e.g., in the 5'-UTR). In some cases, messenger RNAs may contain one or more intronic sequences capable of being excised from the messenger RNA. In some embodiments, a messenger RNA may include a 5' cap structure, a chain terminating nucleotide, a stem loop, a poly A sequence, and / or a polyadenylation signal. Any one of the regions of a messenger RNA may include one or morealternative components (e.g., an alternative nucleoside). For example, the 3'- stabilizing region may contain an alternative nucleoside such as an L-nucleoside, an inverted thymidine, or a 2'-O-methyl nucleoside and / or the coding region, 5'-UTR, 3'- UTR, or cap region may include an alternative nucleoside such as a 5 -substituted uridine (e.g., 5- methoxy uridine), a 1-substituted pseudouridine (e.g., 1-methyl-pseudouridine or 1-ethyl- pseudouridine), and / or a 5-substituted cytidine (e.g., 5-methyl-cytidine).An mRNA of the invention may include an internal ribosome entry site (IRES). An IRES may act as a sole ribosome binding site, or as one of multiple ribosome binding sites. A messenger RNA containing more than one functional ribosome binding site may encode several peptides or polypeptides that are translated independently by the ribosomes. Suitable IRES sequences that may be useful include those from picomaviruses (e.g. FMDV), pest viruses (CFFV), polio viruses (PV), encephalomyocarditis viruses (ECMV), foot-and- mouth disease viruses (FMDV), hepatitis C viruses (HCV), classical swine fever viruses (CSFV), murine leukemia virus (MEV), simian immune deficiency viruses (S1V) and cricket paralysis viruses (CrPV).An mRNA of the invention may comprise a first region of linked nucleosides encoding an antigenic polypeptide, a first flanking region located at the 5'-terminus of the first region (e.g., a 5'-UTR), a second flanking region located at the 3'-terminus of the first region (e.g., a 3'-UTR), at least one 5'-cap region, and a 3'- stabilizing region.Messenger RNA nucleotides may be naturally or non-naturally occurring. The 5'-UTR, (b) the open reading frame (ORF), (c) the 3'-UTR, (d) the poly A tail, and any combination of (a, b, c, or d above) comprise naturally occurring canonical nucleotides A (adenosine), G (guanosine), C (cytosine), U (uridine), or T (thymidine). In some embodiments, the nucleobase is an alternative uracil. Exemplary nucleobases and nucleosides having an alternative uracil include pseudouridine (psi), pyridin-4-one ribonucleoside, 5-aza-uracil, 6- aza-uracil, 2-thio-5-aza-uracil, 2-thio-u raci I (s2U), 4-thio-uracil (s4U), 4-thio-pseudouridine, 2-thio-pseudouridine, 5- hydroxy-uracil, 5-aminoallyl-uracil, 5-halo-uracil (e.g., 5-iodo-uracil or 5-bromo-uracil), 3-methyl-uracil (m3U), 5-methoxy-uracil (mo5U), uracil 5-oxyacetic acid (cmo5U), uracil 5-oxyacetic acid methyl ester (mcmo5U), 5-carboxymethyl-uracil (cm5U), 1 - carboxymethyl-pseudouridine, 5-carboxyhydroxymethyl-uracil (chm5U), 5- carboxyhydroxymethyl-uracil methyl ester (mchm5U), 5-methoxycarbonylmethyl-uracil (mcm5U), 5-methoxycarbonylmethyl-2-thio-uracil (mcm5s2U), 5-aminomethyl- 2-thio-uracil (nm5s2U), 5-methylaminomethyl-uracil (mnm5U), 5-methylaminomethyl- 2-thio-uracil (mnm5s2U), 5-methylaminomethyl-2-seleno-uracil (mnm5se2U), 5-carbamoylmethyl-uracil (ncm5U), 5-carboxymethylaminomethyl-uracil (cmnm5U), 5- carboxymethylaminomethyl-2- thio-uracil (cmnm5s2U), 5-propynyl-uracil, 1-propynyl-pseudouracil, 5-taurinomethyl-uracil (rm5U), 1-taurinomethyl-pseudouridine, 5-taurinomethyl-2-thio-uracil (rm5s2U), 1- taurinomethyl-4-thio-pseudouridine, 5-methyl-uracil (m5U, i.e., having the nucleobase deoxythymine), 1-methyl-pseudouridine, 1-ethyl-pseudouridine (Etly), 5-methyl-2-thio-uracil (m5s2U), 1 -methyl-4-thio-pseudouridine, 4-thio- 1-methyl-pseudouridine, 3-methyl- pseudouridine, 2-thio- 1-methyl-pseudouridine, 1 methyl- 1-deaza-pseudouridine, 2-thio- 1- methyl- 1-deaza-pseudouridine, dihydrouracil (D), dihydropseudouridine, 5,6-dihydrouracil, 5- methyl-dihydrouracil (m5D), 2-thio-dihydrouracil, 2-thio-dihydropseudouridine, 2- methoxy-uracil, 2-methoxy-4-thio-uracil, 4-methoxy-pseudouridine, 4-methoxy-2-thio- pseudouridine, Nl-methyl-pseudouridine, 3-(3-amino-3-carboxypropyl)uracil (acp3U), 1- methyl-3-(3-amino-3-carboxypropyl)pseudouridine, 5-(isopentenylaminomethyl)uracil (inm5U), 5-(isopentenylaminomethyl)-2-thio-uracil (inm5s2U), 5,2'-O-dimethyl-uridine (m5Um), 2-thio-2'-O_methyl-uridine (s2Um), 5- methoxycarbonylmethyl-2'-O-methyl- uridine (mcm5Um), 5-carbamoylmethyl-2'-O- methyl-uridine (ncm5Um), 5- carboxymethylaminomethyl-2'-O-methyl-uridine (cmnm5Um), 3, 2'-O-dimethyl- uridine (m3Um), and 5-(isopentenylaminomethyl)-2'-O-methyl-uridine (inm5Um), 1-thio-uracil, deoxythymidine, 5-(2-carbomethoxyvinyl)-uracil, 5-(carbamoylhydroxymethyl)-uracil, 5- carbamoylmethyl-2-thio-uracil, 5- carboxymethyl-2-thio-uracil, 5-cyanomethyl-uracil, 5- methoxy-2-thio-uracil, and 5-[3-(l-E-propenylamino)]uracil. In one embodiment, the modified uracil is pseudouridine. In one embodiment, the modified uracil is Nl-methyl- pseudouridine.In some embodiments, the mRNA one or more alternative components which impart useful properties including increased stability and / or the lack of a substantial induction of the innate immune response of a cell into which the poly messenger RNA is introduced. For example, an alternative messenger RNA exhibits reduced degradation in a cell into which the messenger RNA is introduced, relative to a corresponding unaltered messenger RNA. These alternative species may enhance the efficiency of protein production, intracellular retention of the messenger RNA, and / or viability of contacted cells, as well as possess reduced immunogenicity.In some embodiments, the nucleobase is an alternative cytosine. Exemplary nucleobases and nucleosides having an alternative cytosine include 5-aza-cytosine, 6-aza-cytosine, pseudoisocytidine, 3-methyl-cytosine (m3C), N4-acetyl-cytosine (ac4C), 5-formyl-cytosine (f5C), N4-methyl-cytosine (m4C), 5-methyl-cytosine (m5C), 5-halo-cytosine (e.g., 5-iodo- cytosine), 5-hydroxymethyl-cytosine (hm5C), 1-methyl-pseudoisocytidine, pyrrolo-cytosine, pyrrolo-pseudoisocytidine, 2-thio-cytosine (s2C), 2-thio-5-methyl-cytosine, 4-thio- pseudoisocytidine, 4-thio-l-methyl-pseudoisocytidine, 4-thio-l-methyl-l-deaza-pseudo isocytidine, 1-methyl- 1-deaza-pseudoisocytidine, zebularine, 5-aza-zebularine, 5-methyl- zebularine, 5-aza-2-thio-zebularine, 2-thio-zebularine, 2-methoxy-cytosine, 2-methoxy-5- methyl-cytosine, 4-methoxy-pseudoisocytidine, 4-methoxy-l-methyl-pseudoisocytidine, lysidine (k2C), 5,2'-O- dimethyl-cytidine (m5Cm), N4-acetyl-2'-O-methyl-cytidine (ac4Cm), N4,2'-O- dimethyl-cytidine (m4Cm), 5-formyl-2'-O-methyl-cytidine (f5Cm), N4,N4,2'-O- trimethyl-cytidine (m42Cm), 1 -thio-cytosine, 5-hydroxy-cytosine, 5-(3-azidopropyl)- cytosine, and 5-(2-azidoethyl)-cytosine. In one example, the modified cytosine is 5-methyl- cytosine.

[0075] In some embodiments, the nucleobase is an alternative adenine. Exemplarynucleobases and nucleosides having an alternative adenine include 2-amino-purine, 2,6- diaminopurine, 2-amino-6-halo-purine (e.g., 2-amino-6-chloro-purine), 6-halo-purine (e.g., 6-chloro-purine), 2-amino-6-methyl-purine, 8-azido-adenine, 7-deaza-adenine, 7-deaza-8- aza-adenine, 7-deaza-2-amino-purine, 7-deaza-8-aza-2-amino-purine, 7-deaza-2,6- diaminopurine, 7-deaza-8-aza-2,6-diaminopurine, 1-methyl-adenine (mlA), 2-methyl- adenine (m2A), N6-methyl-adenine (m6A), 2-methylthio-N6-methyl-adenine (ms2m6A), N6- isopentenyl-adenine (i6A), 2-methylthio-N6-isopentenyl-adenine (ms2i6A), N6-(cis- hydroxyisopentenyl)adenine (io6A), 2-methylthio-N6-(cis-hydroxyisopentenyl)adenine (ms2io6A), N6-glycinylcarbamoyl-adenine (g6A), N6-threonylcarbamoyl-adenine (t6A), N6- methyl-N6-threonylcarbamoyl-adenine (m6t6A), 2-methylthio-N6-threonylcarbamoyl- adenine (ms2g6A), N6,N6-dimethyl-adenine (m62A), N6-hydroxynorvalylcarbamoyl-adenine (hn6A), 2-methylthio-N6-hydroxynorvalylcarbamoyl-adenine (ms2hn6A), N6-acetyl- adenine (ac6A), 7-methyl- adenine, 2-methylthio-adenine, 2-methoxy-adenine, N6,2'-O-dimethyl- adenosine (m6Am), N6,N6,2'-O-trimethyl-adenosine (m62Am), l,2'-O-dimethyl-adenosine (mlAm), 2-amino-N6-methyl-purine, 1-thio-adenine, 8-azido-adenine, N6-(19-amino- pentaoxanonadecyl)-adenine, 2,8-dimethyl-adenine, N6-formyl-adenine, and N6-hydroxy methyl-adenine.In some embodiments, the nucleobase is an alternative guanine. Exemplary nucleobases and nucleosides having an alternative guanine include inosine (I), 1-methyl-inosine (mil), wyosine (imG), methylwyosine (mimG), 4-demethyl-wyosine (imG-14), iso wyo sine (imG2), wybutosine (yW), peroxywybutosine (o2yW), hydroxywybutosine (OHyW), undermodified hydroxywybutosine (OHyW*), 7-deaza-guanine, queuosine (Q), epoxyqueuosine (oQ), galacto syl-queuo sine (galQ), manno syl-queuo sine (manQ), 7-cyano-7-deaza-guanine (preQO), 7-aminomethyl-7-deaza-guanine (preQi), archaeosine (G+), 7-deaza-8-aza-guanine, 6-thio-guanine, 6-thio-7-deaza-guanine, 6-thio-7-deaza-8-aza-guanine, 7-methyl-guanine (m7G), 6- thio-7-methyl-guanine, 7-methyl-inosine, 6-methoxy-guanine, 1-methyl-guanine (mIG), N2-methyl-guanine (m2G), N2,N2-dimethyl-guanine (m22G), N2,7-dimethyl-guanine (m2,7G), N2, N2,7- dimethyl-guanine (m2,2,7G), 8-oxo-guanine, 7-methyl-8-oxo-guanine, 1- methyl-6-thio-guanine, N2-methyl-6-thio-guanine, N2,N2-dimethyl-6-thio-guanine, N2- methyl-2'-O-methyl-guanosine (m2Gm), N2,N2-dimethyl-2'-O-methyl-guanosine (m22Gm), 1- methyl-2'-O-methyl-guanosine (mIGm), N2,N7-dimethyl-2'-O-methyl-guanosine (m2,7Gm), 2'-O-methyl-inosine (Im), l,2'-O-dimethyl-inosine (mllm), 1-thio-guanine, and O- 6-methyl-guanine.The alternative nucleobase of a nucleotide can be independently a purine, a pyrimidine, a purine or pyrimidine analog. For example, the nucleobase can be an alternative to adenine, cytosine, guanine, uracil, or hypoxanthine. In another embodiment, the nucleobase can also include, for example, naturally-occurring and synthetic derivatives of a base, including pyrazolo[3,4-d]pyrimidines, 5-methylcytosine (5-me-C), 5-hydroxymethyl cytosine, xanthine, hypoxanthine, 2-aminoadenine, 6-methyl and other alkyl derivatives of adenine and guanine, 2-propyl and other alkyl derivatives of adenine and guanine, 2-thiou raci I, 2-thiothymine and 2-thiocytosine, 5-propynyl uracil and cytosine, 6-azo uracil, cytosine and thymine, 5-uracil (pseudo uracil), 4-thiouracil, 8-halo (e.g., 8-bromo), 8-amino, 8-thiol, 8- thioalkyl, 8-hydroxy and other 8-substituted adenines and guanines, 5-halo particularly 5- bromo, 5-trifiuoromethyl and other 5-substituted uracils and cytosines, 7-methylguanine and 7-methyladenine, 8- azaguanine and 8-azaadenine, deazaguanine, 7-deazaguanine, 3- deazaguanine, deazaadenine, 7-deazaadenine, 3-deazaadenine, pyrazolo[3,4-d]pyrimidine, imidazo[l,5-a] 1,3,5 triazinones, 9-deazapurines, imidazo[4,5-d]pyrazines, thiazolo[4,5- d]pyrimidines, pyrazin-2-ones, 1,2,4-triazine, pyridazine; or 1,3,5 triazine.An mRNA of the invention may be prepared according to any available technique known in the art. Messenger RNA may be prepared by, for example, enzymatic synthesis which provides a process of template-directed synthesis of RNA molecules from an engineered DNA template comprised of an upstream bacteriophage promoter sequence linked to a downstream sequence encoding the gene of interest. Template DNA can be prepared for in vitro transcription from several sources with appropriate techniques which are well known in the art including, but not limited to, plasmid DNA and polymerase chain reaction amplification.Transcription of the RNA occurs in vitro using the appropriate linearized DNA template in the presence of the corresponding RNA polymerase and adenosine, guanosine, uridine and cytidine ribonucleoside triphosphates (rNTPs) under conditions that support polymerase activity while minimizing potential degradation of the resultant mRNA transcripts. In vitro transcription can be performed using a variety of commercially available kits including, but not limited to RiboMax Large Scale RNA Production System (Promega), MegaScript Transcription kits (Life Technologies) as well as with commercially available reagents including RNA polymerases and rNTPs. The methodology for in vitro transcription of mRNA is well-known in the art.The desired in vitro transcribed messenger RNA is then purified from the undesired components of the transcription or associated reactions. Techniques for the isolation of the messenger RNA transcripts are well known in the art and include phenol / chloroform extraction or precipitation with either alcohol in the presence of monovalent cations or lithium chloride.In some embodiments, the messenger RNA associated with the LNP is a self-amplifying messenger RNA (SAM) molecule. In certain embodiments, the SAM is derived from or based on an alphavirus. Such SAM molecules are known in the art and can be produced using replication elements derived from, for example, alphaviruses substituting the structural viral proteins with a nucleotide sequence encoding a protein of interest. The target cell to which the SAM is delivered generates an exponential increase of encoded gene products, such as proteins or antigens, which can accumulate in the cells or be secreted therefrom. The SAM may contain one or more genes selected from the group consisting of viral replicases, viral proteases, viral helicases and other nonstructural viral proteins, and may also comprise 5'-and 3'-end cis-active replication sequences and, optionally, a heterologous sequence that encodes a desired amino acid sequence. A subgenomic promoter that directs expression of the heterologous sequence may be present. In embodiments, the heterologous sequence may be fused in frame to other coding regions in the SAM and / or be under the control of an internal ribosome entry site (IRES).In some embodiments, the RNA associated with the LNP is a small activating RNA (saRNA), which is a TNA that causes enhancement of endogenous messenger RNA transcription. The saRNA molecules are small double-stranded nucleic acids (See e.g., Voutila et al. Mol Ther. 2017 Dec 6;25(12):2705-2714.The RNA of the invention may encode one or more polypeptide antigens that contain a range of epitopes such as epitopes capable of eliciting either a helper T-cell response or a cytotoxic T-cell response, or both. In some embodiments, the RNA may be engineered to express multiple nucleotide sequences, from two or more open reading frames, thereby allowing co- expression of proteins, such as two or more antigens together with cytokines or other immunomodulators, which can enhance the generation of an immune response. Such a SAM molecule might be useful in the simultaneous production of various proteins as a bivalent or multivalent vaccine.The RNA, including mRNA and self-amplifying RNA may be prepared using any suitable method known in the art. An RNA molecule that contains modified nucleotides can be prepared by transcribing a DNA that encodes the RNA molecule using a suitable DNA- dependent RNA polymerase, such as T7 phage RNA polymerase, SP6 phage RNA polymerase, T3 phage RNA polymerase, and the like, or mutants of the polymerases which allow efficient incorporation of modified nucleotides into RNA. The incorporation of nucleotide analogs into an RNA may be employed to alter the stability of such RNA molecule, to increase resistance against RNases, to establish replication after introduction into appropriate host cells ("infectivity" of the RNA), and / or to induce or reduce innate and adaptive immune responses.Process of making lipid nanoparticlesLNPs of the invention can be made using approaches which are well-known in the art of formulation. For example, suitable LNPs can be formed using mixing processes such as microfluidics, including herringbone micromixing, and T-junction mixing of two fluid streams, one of which contains an RNA, e.g., mRNA, typically in an aqueous solution, and the other of which has the various required lipid components, typically in ethanol.Lipid particles (e.g., lipid nanoparticles) can form spontaneously upon mixing of the nucleic acid, e.g., mRNA, and the lipid(s). Depending on the desired particle size distribution, the resultant nanoparticle mixture can be extruded through a membrane (e.g., 100 nm cut-off) using, for example, a thermobarrel extruder, such as Lipex Extruder (Northern Lipids, Inc). In some cases, the extrusion step can be omitted. Ethanol removal and simultaneous bufferexchange can be accomplished by, for example, dialysis or tangential flow filtration. In one embodiment, the lipid nanoparticles are formed as described in Example 3 described in U.S. Provisional Application No. 63 / 194,620.Methods for preparing LNPs are disclosed, e.g., in W02022 / 261101 WO2019051289, US2013 / 0037977, US2010 / 0015218, US2013 / 0156845, US2013 / 0164400, US2012 / 0225129, US2010 / 0130588, US2007 / 0042031, US2004 / 0142025, Kulkarni et al., 2018, ACS Nano, 12:4787 and Kulkarni et al., 2017, Nanoscale, 36: 133347, the content of each of which is incorporated herein by reference in its entirety. In some embodiments, lipid particles (e.g., lipid nanoparticles) can be prepared using a continuous mixing method, a direct dilution process, or an in-line dilution process. The processes and apparatuses for apparatuses for preparing lipid nanoparticles using direct dilution and in-line dilution processes are described, e.g., in US2007 / 0042031. The processes and apparatuses for preparing lipid nanoparticles using stepwise dilution processes are described in US2004 / 0142025.According to some embodiments, the disclosure provides for an LNP comprising a TNA and an ionizable lipid. For example, a lipid nanoparticle formulation that is made and loaded with a TNA is disclosed in WO2019051289. In one embodiment, the lipid particles (e.g., lipid nanoparticles) can be prepared by an impinging jet process (see e.g., W02022 / 261101). According to some embodiments, the TNA is encapsulated in the lipid(s) thereby protecting it from degradation by a nuclease, e.g., in an aqueous solution. In one embodiment, the TNA in the lipid nanoparticle is not substantially degraded after exposure of the lipid particle to a nuclease at 37°C. for at least about 20, 30, 45, or 60 minutes.The efficiency of encapsulation of the TNA, e.g., mRNA, within the LNPs may be at least 50%, for example about 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99%. In some embodiments, the encapsulation efficiency may be at least 80%. In certain embodiments, the encapsulation efficiency may be at least 90%.Encapsulation of TNA in the lipid nanoparticles can be determined by performing a membrane-impermeable fluorescent dye exclusion assay, which uses a dye that has enhanced fluorescence when associated with nucleic acid, for example, an OLIGREEN® assay or PICOGREEN® assay. Generally, encapsulation is determined by adding the dye to the lipid particle formulation, measuring the resulting fluorescence, and comparing it to the fluorescence observed upon addition of a small amount of non-ionic detergent. Detergent mediated disruption of the lipid bilayer releases the encapsulated TNA, allowing it to interact with the membrane-impermeable dye. Encapsulation of ceDNA can be calculated as E= (Io - 1 ) / lo, where I and Io refer to the fluorescence intensities before and after the addition of detergent, respectively.CompositionsIn one embodiment, the lipid particle formulation is an aqueous solution. In one embodiment, the lipid particle (e.g., lipid nanoparticle) formulation is a lyophilized powder.According to some aspects, the disclosure provides for a lipid particle formulation further comprising one or more pharmaceutical excipients. In one embodiment, the lipid particle (e.g., lipid nanoparticle) formulation further comprises sucrose, Tris buffer, trehalose and / or glycine. Pharmaceutical compositions for therapeutic purposes can be formulated as a solution, microemulsion, dispersion, liposomes, or other ordered structure suitable for high TNA (e.g., mRNA) concentration. Sterile injectable solutions can be prepared by incorporating the TNA (e.g., mRNA) in the required amount in an appropriate buffer (e.g., pharmaceutically acceptable excipient) with one or a combination of ingredients enumerated above, as required, followed by filtered sterilization.In one embodiment, lipid particles (e.g., lipid nanoparticles) are solid core particles that possess at least one lipid bilayer. In one embodiment, the lipid particles (e.g., lipid nanoparticles) have a non-bilayer structure, i.e., a non-lamellar (i.e., non-bilayer) morphology. Without limitations, the non-bilayer morphology can include, for example, three dimensional tubes, rods, cubic symmetries, etc. The non-lamellar morphology (i.e., non-bilayer structure) of the lipid particles (e.g., lipid nanoparticles) can be determined using analytical techniques known to and used by those of skill in the art. Such techniques include, but are not limited to, Cryo-Transmission Electron Microscopy ("Cryo-TEM"), Differential Scanning calorimetry ("DSC"), X-Ray Diffraction, and the like. For example, the morphology of the lipid particles (lamellar vs. non-lamellar) can readily be assessed and characterized using, e.g., Cryo-TEM analysis as described in US2010 / 0130588. In one embodiment, the lipid particles (e.g., lipid nanoparticles) having a non-lamellar morphology are electron dense. In one embodiment, the disclosure provides for a lipid particle (e.g., lipid nanoparticle) that is either unilamellar or multilamellar in structure. In some aspects, the disclosure provides for a lipid particle (e.g., lipid nanoparticle) formulation that comprises multi-vesicular particles and / or foam-based particles. By controlling the composition and concentration of the lipid components, one can control the rate at which the lipid conjugate exchanges out of the lipid particle and, in turn, the rate at which the lipid particle (e.g., lipid nanoparticle) becomes fusogenic. In addition, other variables including, for example, pH, temperature, or ionic strength, can be used to vary and / or control the rate at which the lipid particle (e.g., lipid nanoparticle) becomes fusogenic. By controlling the composition and concentration of the lipid conjugate, one can control the lipid particle size. In one embodiment, the pKa' of formulated cationic lipids can be correlated with the effectiveness of the LNPs for deli very of nucleic acids (see Jayaraman et al., Angewandte Chemie, International Edition (2012), 51(34), 8529-8533; Semple et al., Nature Biotechnology 28, 172-176 (2010), both of which are incorporated by reference in their entireties). In one embodiment, the preferred range of pKa' for the ionizable lipid particle is about 6-7. In one embodiment, the pKa' of the cationic lipid can be determined in lipid particles (e.g., lipid nanoparticles) using an assay based on fluorescence of 2-(p-toluidino)-6-napthalene sulfonic acid (TNS).Pharmaceutical compositions or formulations can optionally comprise one or more additional active substances, e.g., therapeutically and / or prophylactically active substances. Pharmaceutical compositions or formulations of the present invention can be sterile and / or pyrogen-free. General considerations in the formulation and / or manufacture of pharmaceutical agents can be found, for example, in Remington: The Science and Practice of Pharmacy 21st ed., Lippincott Williams & Wilkins, 2005 (incorporated herein by reference in its entirety). In some embodiments, compositions are administered to humans, human patients or subjects. For the purposes of the present disclosure, the phrase "active ingredient" generally refers to a TNA, e.g., mRNA, to be delivered as described herein. Formulations and pharmaceutical compositions described herein can be prepared by any method known or hereafter developed in the art of pharmacology.A pharmaceutical composition or formulation in accordance with the present disclosure can be prepared, packaged, and / or sold in bulk, as a single unit dose, and / or as a plurality of single unit doses. As used herein, a "unit dose" refers to a discrete amount of the pharmaceutical composition comprising a predetermined amount of the active ingredient. The amount of the active ingredient is generally equal to the dosage of the active ingredient that would be administered to a subject and / or a convenient fraction of such a dosage such as, for example, one-half or one-third of such a dosage.Relative amounts of a TNA, the pharmaceutically acceptable excipient, and / or any additional ingredients in a pharmaceutical composition in accordance with the present disclosure can vary, depending upon the identity, size, and / or condition of the subject being treated and further depending upon the route by which the composition is to be administered. The compositions and formulations described herein may contain at least one TNA, such as a mRNA. As a non-limiting example, the composition or formulation can contain 1, 2, 3, 4 or 5 TNAs. In some embodiments, the composition or formulation can comprise a TNA in linear and / or circular form, and in single-stranded and / or doublestranded form. Although the descriptions of pharmaceutical compositions and formulations provided herein are principally directed to pharmaceutical compositions and formulations that are suitable for administration to humans, it will be understood by the skilled artisan that such compositions are generally suitable for administration to any other animal, e.g., to non-human animals, e.g. non-human mammals.Pharmaceutically acceptable excipient, includes, but are not limited to, any and all solvents, dispersion media, or other liquid vehicles, dispersion or suspension aids, diluents, granulating and / or dispersing agents, surface active agents, isotonic agents, thickening or emulsifying agents, preservatives, binders, lubricants or oil, coloring, sweetening or flavoring agents, stabilizers, antioxidants, antimicrobial or antifungal agents, osmolality adjusting agents, pH adjusting agents, buffers, chelators, cyoprotectants, and / or bulking crosslinked polyvinyl pyrrolidone (crospovidone), cellulose, methylcellulose, carboxymethyl cellulose, cross-linked sodium carboxymethyl cellulose (croscarmellose), magnesiumaluminum silicate (V EEG UM®), sodium lauryl sulfate, etc., and / or combinations thereof. Exemplary surface active agents and / or emulsifiers include, but are not limited to, natural emulsifiers ( e.g., acacia, agar, alginic acid, sodium alginate, tragacanth, chondrux, cholesterol, xanthan, pectin, gelatin, egg yolk, casein, wool fat, cholesterol, wax, and lecithin), sorbitan fatty acid esters (e.g., polyoxyethylene sorbitan monooleate [TWEEN®80], sorbitan monopalmitate [SPAN®40], glyceryl monooleate, polyoxyethylene esters, polyethylene glycol fatty acid esters ( e.g., CREMOPHOR®), polyoxyethylene ethers (e.g., polyoxyethylene lauryl ether [BRIJ®30]), PLURONIC® block copolymers, e.g. POLOXAMER® 188, etc. and / or combinations thereof. Exemplary binding agents include, but are not limited to, starch, gelatin, sugars (e.g., sucrose, glucose, dextrose, dextrin, molasses, lactose, lactitol, mannitol), amino acids (e.g., glycine), natural and synthetic gums (e.g., acacia, sodium alginate), ethylcellulose, hydroxyethylcellulose, hydroxypropyl methylcellulose, etc., and combinations thereof.Oxidation is a potential degradation pathway for TNAs, especially for liquid or freeze-dried DNA formulations. To prevent oxidation, antioxidants can be added to the formulations. Exemplary antioxidants include, but are not limited to, alpha tocopherol, ascorbic acid, ascorbyl palmitate, benzyl alcohol, butylated hydroxyanisole, m-cresol, methionine, butylated hydroxytoluene, monothioglycerol, sodium or potassium metabisulfite, propionic acid, propyl gallate, sodium ascorbate, etc., and combinations thereof. Exemplary chelating agents include, but are not limited to, ethylenediaminetetraacetic acid (EDTA), disodium edetate, diethylenetriaminepentaacetic acid (DTPA, in ionized forms), citric acid monohydrate, fumaric acid, malic acid, phosphoric acid, sodium edetate, tartaric acid, Trisodium edetate, etc., and combinations thereof. Exemplary antimicrobial or antifungal agents include, but are not limited to, benzalkonium chloride, benzethonium chloride, methyl paraben, ethyl paraben, propyl paraben, butyl paraben, benzoic acid, hydroxybenzoic acid, potassium or sodium benzoate, potassium or sodium sorbate, sodium propionate, sorbic acid, etc., and combinations thereof. Exemplary preservatives include, but are not limited to, vitamin A, vitamin C, vitamin E, beta-carotene, citric acid, ascorbic acid, butylated hydroxyanisole, ethylenediamine, sodium lauryl sulfate (SLS), sodium lauryl ether sulfate (SLES), p-amino benzoic acid, methyl and / or propyl parabens, etc., and combinations thereof. In some embodiments, the pH of polynucleotide solutions is maintained between pH 5 and pH 8 to improve stability. Exemplary buffers to control pH can include, but are not limited to sodium phosphate, sodium citrate, sodium succinate, histidine (or histidine HCI), sodium malate, sodium carbonate, etc., and / or combinations thereof. Exemplary lubricating agents include, but are not limited to, magnesium stearate, calcium stearate, stearic acid, silica, talc, malt, hydrogenated vegetable oils, polyethylene glycol, sodium benzoate, sodium or magnesium lauryl sulfate, etc., and combinations thereof. The pharmaceutical composition or formulation described here can contain a cryoprotectant to stabilize a polynucleotide described herein during freezing. Exemplary cryoprotectants include, but are not limited to mannitol, sucrose, trehalose, lactose,glycerol, dextrose, etc., and combinations thereof. The pharmaceutical composition or formulation described here can contain a bulking agent in lyophilized polynucleotide formulations to yield a "pharmaceutically elegant" cake, stabilize the lyophilized polynucleotides during long term (e.g., 36-month) storage. Exemplary bulking agents of the present invention can include, but are not limited to sucrose, trehalose, mannitol, glycine, lactose, raffinose, and combinations thereof. In some embodiments, the pharmaceutical composition or formulation further comprises a delivery agent. The delivery agent of the present disclosure can include, without limitation, liposomes, lipid nanoparticles, lipidoids, polymers, lipoplexes, microvesicles, exosomes, extracellular vesicles, peptides, proteins, cells transfected with polynucleotides, hyaluronidase, nanoparticle mimics, nanotubes, conjugates, and combinations thereof.Unit DosageIn one embodiment, the pharmaceutical compositions can be presented in unit dosage form. A unit dosage form will typically be adapted to one or more specific routes of administration of the pharmaceutical composition. In some embodiments, the unit dosage form is adapted for intravenous, intramuscular, or subcutaneous administration. In some embodiments, the unit dosage form is adapted for intrathecal or intracerebroventricular administration. In some embodiments, the unit dosage form is adapted for administration by inhalation. In some embodiments, the unit dosage form is adapted for administration by a vaporizer. In some embodiments, the unit dosage form is adapted for administration by a nebulizer. In some embodiments, the unit dosage form is adapted for administration by an aerosolizer. In some embodiments, the unit dosage form is adapted for oral administration, for buccal administration, or for sublingual administration. In some embodiments, the pharmaceutical composition is formulated for topical administration. The amount of active ingredient which can be combined with a carrier material to produce a single dosage form will generally be that amount of the compound which produces a therapeutic effect. According to some embodiments, the LNP / TNA is for administration at a dose of about 0.02 pg to about 50 mg, about 0.02 pg to about 0.2 pg, or about 0.2 pg to about 2.0 pg, about 1 pg to about 25 pg, about 25 pg to about 50 pg, about 50 pg to about 100 pg, about 100 pg to about 200 pg, about 200 pg to about 300 pg, about 300 pg to about 400 pg, about 400 pg to about 500 pg, about 500 pg to about 750 pg, about 750 pg to about 1.0 mg, about 1 mg to about 10 mg, about 10 mg to about 25 mg, about 25 mg to about 50 mg.Methods of UsingThe pharmaceutical compositions comprising a lipid nanoparticle (LN P) and a therapeutic nucleic acid (TNA) can be used to introduce a nucleic acid sequence (e.g., a TNA) into a host cell. In one embodiment, the host cell is in vitro. In one embodiment, the host cell is in vivo. According to some embodiments, the subject is a human. In one embodiment, introduction of a nucleic acid sequence in a host cell using the pharmaceutical compositions comprising alipid nanoparticle (LN P) and a therapeutic nucleic acid (TNA), as described herein, can be monitored with appropriate biomarkers from treated patients to assess gene expression.Provided herein are methods of treating a disease or disorder in a subject comprising introducing into a cell in need thereof (for example, a muscle cell or tissue, or other affected cell type) of the subject a therapeutically effective amount of pharmaceutical composition comprising a lipid nanoparticle (LNP) and a therapeutic nucleic acid (TNA), wherein the LNP comprises an ApoE polypeptide, or a fragment thereof and / or an ApoB polypeptide, or a fragment thereof, linked to the LNP. While the TNA lipid nanoparticles can be introduced in the presence of a carrier, such a carrier is not required. Provided herein are methods for providing a subject in need thereof with a diagnostically- or therapeutically- effective amount of the pharmaceutical composition comprising a LNP and a TNA.In general, the pharmaceutical composition comprising an LNP and a TNA can be used to deliver any TNA in accordance with the description above to treat, prevent, or ameliorate the symptoms associated with any disorder related to gene expression. Illustrative disease states include, but are not-limited to: cystic fibrosis (and other diseases of the lung), hemophilia A, hemophilia B, thalassemia, anemia and other blood disorders, AIDS, Alzheimer's disease, Parkinson's disease, Huntington's disease, amyotrophic lateral sclerosis, epilepsy, and other neurological disorders, cancer, diabetes mellitus, muscular dystrophies (e.g., Duchenne, Becker), Hurler's disease, adenosine deaminase deficiency, metabolic defects, retinal degenerative diseases (and other diseases of the eye), mitochondriopathies (e.g., Leber's hereditary optic neuropathy (LHON), Leigh syndrome, and subacute sclerosing encephalopathy), myopathies (e.g., facioscapulohumeral myopathy (FSHD) and cardiomyopathies), diseases of solid organs (e.g., brain, liver, kidney, heart), and the like. In some embodiments, the ceDNA vectors as disclosed herein can be advantageously used in the treatment of individuals with metabolic disorders (e.g., ornithine transcarbamoylase deficiency). In one embodiment, the pharmaceutical composition comprising a LNP and a TNA can be used to treat, ameliorate, and / or prevent a disease or disorder caused by mutation in a gene or gene product (i.e., a genetic disorder) include, but are not limited to, metabolic diseases or disorders (e.g., Fabry disease, Gaucher disease, phenylketonuria (PKU), glycogen storage disease); urea cycle diseases or disorders (e.g., ornithine transcarbamoylase (OTC) deficiency); lysosomal storage diseases or disorders (e.g., metachromatic leukodystrophy (MLD), mucopolysaccharidosis Type II (MPSII; Hunter syndrome)); liver diseases or disorders (e.g., progressive familial intrahepatic cholestasis (PFIC); blood diseases or disorders (e.g., hemophilia (A and B), thalassemia, and anemia); cancers and tumors, and genetic diseases or disorders (e.g., cystic fibrosis). According to some embodiments, the genetic disorder is hemophilia A, hemophilia B, phenylketonuria (PKU, Gaucher disease Types I, II and III, Stargardt macular dystrophy, Leber congenital amaurosis (LCA), Usher syndrome, wet AMD. In one embodiment, the pharmaceutical composition comprising an LNP and a TNA may be employed to deliver a heterologous nucleotide sequence, e.g., to correct an abnormal level and / or function of a gene product,such as an absence of, or a defect in, a protein, that results in the disease or disorder. The TNA in lipid nanoparticles as described herein can produce a functional protein and / or modify levels of the protein to alleviate or reduce symptoms resulting from, or confer benefit to, a particular disease or disorder caused by the absence or a defect in the protein. For example, the TNA may be used for producing a functional protein or increased expression of a protein, such as OTC enzyme, Factor VIII, Factor IX, and Factor X, phenylalanine hydroxylase enzyme, alpha galactosidase or beta glucocerebrosidase, arylsulfatase A, iduronate-2-sulfatase, cystic fibrosis transmembrane conductance regulator, G6Pase enzyme, ATP8B 1, ABCB 11, ABCB4, or TJP2. In one embodiment, exemplary TNA encode a protein selected from: lysosomal enzymes (e.g., hexosaminidase A, iduronate sulfatase, associated, erythropoietin, angiostatin, endostatin, superoxide dismutase, globin, leptin, catalase, tyrosine hydroxylase, as well as cytokines (e.g., a interferon, b-interferon, interferon-gamma, interleukin-2, interleukin-4, interleukin 12, granulocyte- macrophage colony stimulating factor, lymphotoxin, and the like), peptide growth factors and hormones (e.g., somatotropin, insulin, insulin-like growth factors 1 and 2, platelet derived growth factor (PDGF), epidermal growth factor (EGF), fibroblast growth factor (FGF), nerve growth factor (NGF), neurotrophic factor-3 and 4, brain-derived neurotrophic factor (BDNF), glial derived growth factor (GDNF), transforming growth factor-a and -b, and the like), receptors (e.g., tumor necrosis factor receptor). In some exemplary embodiments, the transgene encodes a monoclonal antibody specific for one or more desired targets. In some exemplary embodiments, the antibody may be a full-length antibody or antibody fragment, e.g., antigen binding fragment, thereof.AdministrationIn one embodiment, the pharmaceutical compositions comprising a lipid nanoparticle (LNP) and a therapeutic nucleic acid (TNA), as described herein, can be administered to an organism for transduction of cells in vivo. In one embodiment, the TNA can be administered to an organism for transduction of cells ex vivo. Generally, administration is by any of the routes normally used for introducing a molecule into ultimate contact with blood or tissue cells. Suitable methods of administering such nucleic acids are available and well known to those of skill in the art, and, although more than one route can be used to administer a particular composition, a particular route can often provide a more immediate and more effective reaction than another route. Exemplary modes of administration of the pharmaceutical composition of the invention include oral, rectal, buccal (e.g., sublingual), transmucosal (regardless of anatomical location), parenteral , including, but not limited to intravenous, intraarterial, subcutaneous, intradermal, intracranial, intramuscular (including administration to skeletal, diaphragm and / or cardiac muscle], intrapleural, intracerebral, intraosseous (including bone marrow), and intraarticular, intranasal, inhalation (e.g., via an aerosol), vaginal, intrathecal, intraocular, transdermal, intraendothelial, in utero (or in ovo), topical (e.g., to both skin and mucosal surfaces, including airway surfaces, and transdermal administration), intralymphatic, intranodal and the like, as well as direct tissue or organinjection (e.g., to liver, eye, skeletal muscle, cardiac muscle, diaphragm muscle or brain). Administration of the pharmaceutical composition can be to any site in a subject, including, without limitation, a site selected from the group consisting of the brain, a skeletal muscle, a smooth muscle, the heart, the diaphragm, the airway epithelium, the liver, the kidney, the spleen, the pancreas, the skin, and the eye. The pharmaceutical composition can be administered to skeletal muscle includes but is not limited to administration to skeletal muscle in the limbs (e.g., upper arm, lower arm, upper leg, and / or lower leg), back, neck, head (e.g., tongue), thorax, abdomen, pelvis / perineum, and / or digits, by intravenous administration, intra-arterial administration, intraperitoneal administration, limb perfusion, (optionally, isolated limb perfusion of a leg and / or arm; see, e.g., Arruda et al. 2005, Blood 105: 3458-3464), and / or direct intramuscular injection. In one embodiment, pharmaceutical composition is administered to cardiac muscle, including left atrium, right atrium, left ventricle, right ventricle and / or septum, e.g., by intravenous administration, intra-arterial administration such as intra-aortic administration, direct cardiac injection (e.g., into left atrium, right atrium, left ventricle, right ventricle), and / or coronary artery perfusion. Administration to diaphragm muscle can be by any suitable method including intravenous administration, intra-arterial administration, and / or intra-peritoneal administration. Administration to smooth muscle can be by any suitable method including intravenous administration, intra-arterial administration, and / or intra-peritoneal administration. In one embodiment, administration can be to endothelial cells present in, near, and / or on smooth muscle. In one embodiment, pharmaceutical composition comprising is administered to the CNS (e.g., to the brain or to the eye). The pharmaceutical composition may be introduced into the spinal cord, brainstem (medulla oblongata, pons), midbrain (hypothalamus, thalamus, epithalamus, pituitary gland, substantia nigra, pineal gland), cerebellum, telencephalon (corpus striatum, cerebrum including the occipital, temporal, parietal and frontal lobes, cortex, basal ganglia, hippocampus and porta amygdala), limbic system, neocortex, corpus striatum, cerebrum, and inferior colliculus. The pharmaceutical compositions may also be administered to different regions of the eye such as the retina, cornea and / or optic nerve., e.g., via subretinal injection, suprachoroidal injection, or intravitreal injection The pharmaceutical composition may be delivered into the cerebrospinal fluid (e.g., by lumbar puncture). The pharmaceutical composition may be administered to the desired region(s) of the CNS by any route known in the art, including but not limited to, intrathecal, intra-ocular, intracerebral, intraventricular, intravenous (e.g., in the presence of a sugar such as mannitol), intranasal, intra-aural, intra-ocular (e.g., intra- vitreous, sub-retinal, anterior chamber) and peri-ocular (e.g., sub-Tenon's region) delivery as well as intramuscular delivery with retrograde delivery to motor neurons. In one embodiment, repeat administrations of the therapeutic product can be made until the appropriate level of expression has been achieved. Thus, in one embodiment, a therapeutic nucleic acid can be administered and re-dosed multiple times.ExamplesSynthesis of amino lipids is carried out with standard methods of organic synthetic methodology. All cited sources, for example, references, publications, databases, database entries, and synthesis art cited herein, are incorporated into this application by reference, even if not expressly stated in the citation. All solvents and reagents are / were obtained commercially and used as such unless noted otherwise.Synthesis of decyl 4-(((4-amino-2,6,7-trioxabicyclo[2.2.2]octan-l-yl)methyl)(4- (heptadecan-9-yloxy)-4-oxobutyl)amino)butanoate (52).To a cold solution of 4-bromobutanoic acid (52-1, 5.0 g, 30 mmol) in dichloromethane (20 mL) was added oxalyl chloride (3.85 mL, 45 mmol) followed by dimethylformamide (1 drop). The resultant mixture was stirred at 0 °C for 2 h and the solvent was then evaporated in a rotary evaporator. The residue was dissolved in dichloromethane (20 mL), cooled to 0 °C, and decan-9-ol (52-2, 7.1 g, 45 mmol) was added followed by pyridine (4.84 mL, 60 mmol). The resultant mixture was stirred at ambient temperature overnight. The mixture was quenched with ice, extracted with dichloromethane (2 x 25 mL). The combined organic layer was washed with water, brine, dried over anhydrous sodium sulfate and evaporated under vacuo. The crude product was purified by silica gel flash chromatography with 0-10% ethyl acetate-dichloromethane to obtain decyl 4-bromobutanoate (52-3, 8.09 g, 88%).1H NMR (400 MHz, CDCI3, ppm): δ 4.04-4.07 (m, 2H), 3.43-3.48 (m, 2H), 2.45-2.49 (m, 2H), 2.09- 2.18 (m, 2H), 1.45-1.6 (m, 2H), 1.24-1.31 (m, 14H), 0.84-0.89 (m, 3H).To a cold solution of 4-bromobutanoic acid (52-1, 5.0 g, 30 mmol) in dichloromethane (20 mL) was added oxalyl chloride (3.85 mL, 45 mmol) followed by dimethylformamide (1 drop). The resultant mixture was stirred at 0°C for 2 h and the solvent was then evaporated in a rotary evaporator. The residue was dissolved in dichloromethane (20 mL), cooled to 0 °C, and heptadecan-9-ol (52-4, 11.5 g, 45 mmol) was added followed by pyridine (4.84 mL, 60 mmol). The resultant mixture was stirred at ambient temperature overnight. The mixture was quenched with ice, extracted with dichloromethane (2 x 25 mL). The combined organic layer was washed with water, brine, dried over anhydrous sodium sulfate and evaporated under vacuo. The crude product was purified by silica gel flash chromatography with 0-10% ethyl acetate-dichloromethane to obtain heptadecan-9-yl 4-bromobutanoate (52-5,11.35 g, 94%).1H NMR (400 MHz, CDCI3, ppm): δ 4.82-4.92 (m, 1H), 3.45-3.62 (m, 2H), 2.47-2.52 (m, 2H), 2.05-2.18 (m, 2H), 1.45-1.6 (m, 4H), 1.21-1.31 (m, 24H), 0.86-0.89 (m, 6H).Synthesis of heptadecan-9-yl 4-(benzylamino)butanoate (Compound 52-6)A mixture of heptadecan-9-yl 4-bromobutanoate (52-3, 3.75 g, 9.27 mmol) and benzylamine (4.97 g, 46.38 mmol) in ethanol (100 mL) was refluxed at 80 °C for 48 h. The solvent was removed, and the product was purified by silica gel flash chromatography with 0-10% ethyl acetate-dichloromethane to obtain the desired compound, with 0-10% ethyl acetatedichloromethane to obtain heptadecan-9-yl 4-(benzylamino)butanoate (52-6, 2.45 g, 88%). The1H NMR showed some impurities, and this product was used in the next step without further purification.1H NMR (400 MHz, CDCI3, ppm): δ 7.22-7.34 (m, 5H), 4.82-4.88 (m, 1H), 4.44 (br s, 1H), 3.77 (s, 2H), 3.45-3.62 (m, 2H), 2.63-2.67 (m, 2H), 2.33-2.37 (m, 2H), 1.80- 1.86 (m, 2H), 1.48-1.52 (m, 2H), 1.21-1.31 (m, 24H), 0.84-0.88 (m, 6H).Synthesis of decyl 4-(benzyl(4-(heptadecan-9-yloxy)-4-oxobutyl)amino)butanoate (52-7)To a solution of heptadecan-9-yl 4-(benzylamino)butanoate (52-6, 500 mg, 1.16 mmol) and decyl 4-bromobutanoate ,(52-3,532 mg, 1.74 mmol) in acetonitrile (100 mL) was added potassium carbonate (480 mg, 3.5 mmol) and potassium iodide (96 mg, 0.6 mmol) and the reaction was heated at 180 °C for 48 h. The reaction mixture was filtered, washed with methylene chloride and the solvents evaporated. Another batch with 2.0 g of heptadecan-9- yl 4-(benzylamino)butanoate (52-6 was carried out by following the above procedure and both the crude mixtures were combined and purified by silica gel flash chromatography using 0-30% ethyl acetate-dichloromethane to obtain decyl 4-(benzyl(4-(heptadecan-9- yloxy)-4-oxobutyl)amino)butanoate (52-7, 2.35 g, 61%). The1H NMR showed some impurities, and this product was used in the next step without further purification.1H NMR (400 MHz, CDCI3, ppm): δ 7.21-7.29 (m, 5H), 4.81-4.87 (m, 1H), 3.98-4.03 (m, 2H), 3.54 (s, 2H), 2.28-2.43 (m, 8H), 1.75-1.81 (m, 4H), 1.45-1.53 (m, 4H), 1.18-1.45 (m, 40H), 0.85-0.88 (m, 9H). APCI: m / z 658.6Synthesis of decyl 4-((4-(heptadecan-9-yloxy)-4-oxobutyl)amino)butanoate (52-8)A solution of decyl 4-(benzyl(4-(heptadecan-9-yloxy)-4-oxobutyl)amino)butanoate (36-7, 2.35 g, 3.6 mmol) in ethyl acetate (100 mL) was stirred overnight under hydrogen atmosphere in the presence of 10% Pd / C (wet, 500 mg). The reaction mixture was filtered through celite rinsing with ethyl acetate and dichloromethane. The filtrate was evaporated to obtain the desired compound, decyl 4-((4-(heptadecan-9-yloxy)-4- oxobutyl)amino)butanoate (52-8, 1.5 g, 75%). (400H M NHMz,R CDCI3, ppm): δ 4.84-4.87 (m, 1H), 4.01-4.05 (m, 2H), 3.62-3.64 (m, 1H), 2.61-2.65 (m, 2H), 2.28-2.40 (m, 4H), 1.75-1.88 (m, 4H), 1.50-1.65 (m, 6H), 1.19-1.43 (m, 40H), 0.83-0.89 (m, 9H). APCI: m / z 568.5Synthesis of decyl 4-((cyanomethyl)(4-(heptadecan-9-yloxy)-4-oxobutyl)amino)butanoate (52-9) and heptadecan-9-yl 4-(2-oxopyrrolidin-l-yl)butanoateTo a solution of decyl 4-((4-(heptadecan-9-yloxy)-4-oxobutyl)amino)butanoate (52-8, 1.45 g, 2.55 mmol) and chloroacetonitrile (289 mg, 3.83 mmol) in acetonitrile (50 mL) was added potassium carbonate (1.06 g, 7.66 mmol) and potassium iodide (212 mg, 1.3 mmol) and the reaction was heated at 180 °C for 48 h. The reaction mixture was filtered, washed with methylene chloride and the solvents evaporated. The crude was purified by silica gel flash chromatography using 0-30% ethyl acetate-dichloromethane to obtain decyl 4- ((cyanomethyl)(4-(heptadecan-9-yloxy)-4-oxobutyl)amino)butanoate (52-9, 148 mg, 10%) and heptadecan-9-yl 4-(2-oxopyrrolidin-l-yl)butanoate (750 mg, 72%). decyl 4-((cyanomethyl)(4-(heptadecan-9-yloxy)-4-oxobutyl)amino)butanoate (52-9):1H NMR (400 MHz, CDCI3, ppm): δ 4.83-4.89 (m, 1H), 4.04-4.07 (m, 2H), 3.56 (s, 2H), 2.53-2.56 (m, 4H), 2.30-2.35 (m, 4H), 1.50-1.78 (m, 8H), 1.19-1.36 (m, 40H), 0.84-0.89 (m, 9H). heptadecan-9-yl 4-(2-oxopyrrolidin-l-yl)butanoate: (410H0 N MMHRz, CDCI3, ppm): 64.84- 4.89 (m, 1H), 3.29-3.42 (m, 4H), 2.29-2.38 (m, 4H), 1.99-2.08 (m, 2H), 1.80-1.90 (m, 2H), 1.65-1.47(m, 2H), 1.45-1.55 (m, 2H), 1.20-1.32 (m, 24H), 0.86-0.89 (m, 6H).Synthesis of decyl 4-(((trimethoxy)methyl)(4-(heptadecan-9-yloxy)-4- oxobutyl)amino)butanoate (Compound 52-10)The acetonitrile-substituted tertiary amine is treated with HCI gas in methanol-containing solvent. The orthoester is isolated from unreacted material and any methyl esters side product occurring by transesterification.Synthesis of decyl 4-((2-(2-amino-3-hydroxy-2-(hydroxymethyl)propoxy)-2-oxoethyl)(4 - (heptadecan-9-yloxy)-4-oxobutyl)amino)butanoate (Compound 52)The decyl 4-(((trimethoxy)methyl)(4-(heptadecan-9-yloxy)-4-oxobutyl)amino)butanoate (52- 10) is treated with Tris(hydroxymethyl)nitromethane to form the bicyclic system with a nitro group opposite to the lipid tails. The resulting nitro-bicyclo lipid compound is reduced with hydrogen gas over PtO2 to produce the desired amino-bicyclo lipid (Compound 36). The amino lipid is purified by column chromatography over alumina, taking care to purify out the unreacted nitro analog.Acidic hydrolysis of bicyclic ring system in Compound 36 is accomplished without hydrolysis of the carboxyl esters of the tails by mild acid treatment to obtain Compound 52.Synthetic Scheme for Compound 51, octyl 10-(2-hydroxyethyl)-16-octyl-14-oxo-7, 13,15- trioxa-10-azatetracosanoate, is shown below.Compound 51Synthesis of 6-(2-(benzyloxy)ethoxy)hexan-l-ol (51-3 in scheme for Compound 51)Chemical Formula: C15H24O3Exact Mass: 252.17Molecular Weight: 252.35To a suspension of sodium hydride (3.94 g, 60% dispersion in oil, 98 mmol) in THF (200 mL) was added dropwise a solution of 2-(benzyloxy)ethan-l-ol (51-2, 10.0 g, 65 mmol) in THF (50 mL) at 0 °C. It was stirred at 0 °C for 1 h. Then a solution of 6-bromohexan-l-ol (51-1, 14.3 g, 78 mmol) was added and the reaction mixture stirred at room temperature overnight. It was quenched with ice, extracted with hexane (3 X 100 mL). The organic layer was washed with brine, dried with anhydrous sodium sulfate and the solvent removed to get the crude compound. The crude compound was purified by flash chromatography on silica gel using 40% ethyl acetate in hexane to get pure 6-(2-(benzyloxy)ethoxy)hexan-l-ol (51-3, 13.9 g, 84%.1H NMR (400 MHz, CDCI3, ppm): δ 7.26-7.35 (m, 5H), 4.57 (s, 2H), 3.39-3.65 (m, 8H), 3.80 (m, 1H), 1.36-1.58 (m, 8H).Synthesis of 6-(2-(benzyloxy)ethoxy)hexanoic acid (51-4 in scheme for Compound 51)Chemical Formula: C15H22O4 Exact Mass: 266.15 Molecular Weight: 266.34To a solution of 6-(2-(benzyloxy)ethoxy)hexan-l-ol (51-3, 4.0 g, 16 mmol) in acetone (100 mL) at 0 °C, was added Jones reagent (10.0 mL, 2M, 32 mmol). The reaction mixture was stirred at 0 °C for 3h. The reaction was quenched with isopropanol. The solvent was then removed, and the crude product taken in dichloromethane (100 mL). the organic layer was washed with sodium hydroxide (IN, 3X50 mL). The aqueous part was then acidified with hydrochloric acid (IN) and extracted with ethyl acetate. The ethyl acetate layer was washed with brine, dried over anhydrous sodium sulfate and solvent removed to give the product, 6-(2-(benzyloxy)ethoxy) hexanoic acid (51-4, 2.4 g), which was directly used in the next step.Synthesis of octyl 6-(2-(benzyloxy)ethoxy)hexanoate (51-6 in scheme for Compound 51)Chemical Formula: C23H38O4 Exact Mass: 378.28 Molecular Weight: 378.55To a solution of 6-(2-(benzyloxy)ethoxy) hexanoic acid (51-4, 2.1 g, 8 mmol) in dichloromethane (50 mL) at 0 °C, was added oxalyl chloride (1.54 g, 1.06 mL, 12.1 mmol) followed by / V, / V-Dimethylformamide (0.05 mL) under nitrogen atmosphere. The reaction mixture was stirred at 0 °C for 2 h. The solvent was removed, and the crude reaction mixture dried in rotavapor for 2 h. Then the crude acid chloride was dissolved in dichloromethane (50 mL) and octan-l-ol (51-5, 1.58 g, 1.9 mL, 12.1 mmol) was added at 0 °C followed by pyridine (1.28 g, 1.31 mL, 16.1 mmol). The reaction mixture was stirred at room temperature for 18 h under nitrogen. The reaction mixture was quenched with ice. The organic layer was separated washed with brine, dried with anhydrous sodium sulfate and the solvent removed to get the crude product which was purified by flash chromatography to provide octyl 6-(2-(benzyloxy)ethoxy)hexanoate (51-6, 650 mg, 21%) as a colorless oiI.1H NMR (400 MHz, CDCI3, ppm): δ 7.26-7.35 (m, 5H), 4.57 (s, 2H), 4.05 (t, 2H, J 6.8 Hz), 3.60 (s, 4H), 3.46 (t, 2H, J 6.8 Hz), 2.30 (t, 2H, J 6.6 Hz), 1.59-1.72 (m, 8H), 1.26-1.42 (m, 10H), 0.88 (t, 3H, J 6.8 Hz).Synthesis of octyl 6-(2-hydroxyethoxy)hexanoate (51-7 in scheme for Compound 51)Chemical Formula: C16H32O4 Exact Mass: 288.23 Molecular Weight: 288.43To a solution of octyl 6-(2-(benzyloxy)ethoxy)hexanoate (51-6, 1.75 g, 4.62 mmol) in ethyl acetate (50 mL) was added palladium hydroxide on carbon (20%) (325 mg, 0.46 mmol)and the reaction mixture stirred at room temperature under hydrogen atmosphere for 18 h. The reaction mixture was filtered through a bed of celite, washed with ethyl acetate and dichloromethane and the solvent removed to get the crude product octyl 6-(2- hydroxyethoxy)hexanoate (51-7, 1.33 g, 99 %) in quantitative yield. The crude product was carried forward to the next reaction.1H NMR (400 MHz, CDCI3, ppm): δ 4.05 (t, 2H, J 6.8 Hz), 3.73 (t, 2H, J 4.5 Hz), 3.53 (t, 2H, J 4.4 Hz), 3.48 (t, 2H, J 6.4 Hz), 2.31 (t, 2H, J 7.2 Hz), 1.58- 1.70 (m, 6H), 1.20-1.42 (m, 13H), 0.88 (t, 3H, J 6.8 Hz).Synthesis of octyl 6-(2-bromoethoxy)hexanoate (51-8 in scheme for Compound 51)Chemical Formula: C-ieHsiBrOa Exact Mass: 350.15 Molecular Weight: 351.33To a solution of octyl 6-(2-hydroxyethoxy)hexanoate (51-7, 1.33 g, 4.61 mmol) in dichloromethane (100 mL) at room temperature was added carbon tetrabromide (2.14 g, 6.46 mmol) followed by triphenylphosphine (1.69 g, 6.46 mmol). The reaction mixture was stirred at room temperature for 2 h. Solid silica was added to the reaction mixture and the solvent removed. The silica was directly loaded and purified by flash chromatography to get octyl 6-(2-bromoethoxy)hexanoate (51-8, 1.299 g, 80 %) as colorless oil. (400 MHz,1H NMR CDCI3, ppm): δ 4.05 (t, 2H, J 6.6 Hz), 3.73 (t, 2H, J 6.2 Hz), 3.44-3.52 (m, 4H), 2.31 (t, 2H, J 7.6 Hz), 1.57-1.72 (m, 6H), 1.21-144 (m, 12H), 0.88 (t, 3H, J 6.6 Hz).Synthesis of octyl 6-(2-((2-hydroxyethyl)amino)ethoxy)hexanoate (51-10 in scheme for Compound 51)To a solution of octyl 6-(2-bromoethoxy)hexanoate (51-8, 1.29 g, 3.67 mmol) in ethanol (50 mL) was added ethanolamine (51-9, 4.49 g, 4.44 mL, 73.4 mmol) and the reaction mixture refluxed for 18 h at 80 °C. The solvent was removed, and the crude reaction mixture purified by flash chromatography to obtain octyl 6-(2-((2-hydroxyethyl)amino)ethoxy)hexanoate (51- 10, 1.01 g, 83 %) as an yellow oil.The crude compound NMR is below, the material is carried forward to be purified in the next step.1H NM (4R00 MHz, CDCI3, ppm): δ 4.05 (t, 2H, J 6.8 Hz), 3.36-3.58 (m, 6H), 2.79 (t, 4H, J 5.2 Hz), 2.31 (t, 2H, J 7.4 Hz), 2.07 (bs, 2H), 1.58-1.66 (m, 6H), 1.20-1.39 (m, 12H), 0.88 (t, 3H, J 6.8 Hz).Synthesis of 2-bromoethyl heptadecan-9-yl carbonate (15 in scheme for Compound 51)To a solution of 2-bromoethan-l-ol (51-11, 3.00 g, 24.0 mmol) in dichloromethane (50 mL) was added 4-nitrophenyl carbonochloridate (51-12, 5.80 g, 29.0 mmol) followed by DMAP (0.58 g, 5.0 mmol) and pyridine (3.80 g, 2.15 mL, 48.0 mmol) and the reaction mixture stirred at room temperature for 1 h. Then, heptadecan-9-ol (51-14, 18.50 g, 72.0 mmol) was added followed by diisopropylethylamine (9.30 g, 12.5 mL, 72.0 mmol) and the reaction mixture stirred at room temperature overnight. The reaction was quenched with sodium carbonate (IM), washed with water and the solvent removed to get the crude product (51- 15) which was used without further purification.Synthesis of octyl 10-(2-hydroxyethyl)-16-octyl-14-oxo-7,13,15-trioxa-10- azatetracosanoate (Compound 51)To a solution of octyl 6-(2-((2-hydroxyethyl)amino)ethoxy)hexanoate (51-10, 400 mg, 1.21 mmol) in acetonitrile (50 mL) was added 2-bromoethyl heptadecan-9-yl carbonate (51-15, 737 mg, 1.81 mmol) followed by potassium carbonate (500 mg, 3.62 mmol) and potassium iodide (100 mg, 0.60 mmol). The reaction mixture was then heated to reflux at 98 °C for 48 h. The reaction mixture initially turned canary yellow and then slowly the color changes and becomes colorless. The solvent was removed, and the crude compound purified by flash chromatography, followed by reverse phase chromatography using acetonitrile and water in 0.1% trifluoroacetic acid. The pure product was dissolved in dichloromethane (5.0 mL) and solid sodium bicarbonate (100 mg, 1.19 mmol) was added and stirred for 1 hour. It was then filtered, and the solvent removed to get the pure product octyl 10-(2-hydroxyethyl)-16- octyl-14-oxo-7,13,15-trioxa-10-azatetracosanoate (Compound 51 150 mg, 18%) as colorless oil.1H NMR (400 MHz, CD3OD, ppm): δ 4.70-4.73 (m, 1H), 4.51 (t, 2H, J 4.8 Hz), 4.05 (t, 2H, J 6.6 Hz), 3.88 (t, 2H, J 5.2 Hz), 3.79 (t, 2H, J 4.8 Hz), 3.69 (t, 2H, J 4.8 Hz), 3.51-3.54 (m 4H),3.44 (m, 2H), 2.33 (t, 2H, J 7.4 Hz), 1.58-1.65 (m 9H), 1.29-1.41 (m, 38H), 0.88-0.91 (m, 9H). APCI: m / z 658.5, HPLC-ELSD purity = >99%. HPLC-CAD purity = >97%.Synthetic Scheme for Compound 53, 2-((2-((3-(heptadecan-9-yloxy)-3- oxopropanoyl)oxy)ethyl)(2-hydroxyethyl)amino)ethyl octyl adipate:Compound 53Synthesis of benzyl octyl adipate (53-2 in scheme for Compound 53)Chemical Formula: C21H32O4Exact Mass: 348.230 Molecular Weight: 348.48To a solution of benzyl adipate (53-1, 2.1 g, 8 mmol) in dichloromethane (50 mL) at 0 °C was added oxalyl chloride (1.54 g, 1.06 mL, 12.1 mmol) followed by / V, / V-Dimethylformamide (0.05 mL) under nitrogen atmosphere. The reaction mixture was stirred at 0 °C for 2 h. The solvent was removed, and the crude reaction mixture dried in rotavapor for 2 h. Then the crude acid chloride was dissolved in dichloromethane (50 mL) and octan-l-ol (51-5, 1.58 g, 1.9 mL, 12.1 mmol) was added at 0 °C followed by pyridine (1.28 g, 1.31 mL, 16.1 mmol). The reaction mixture was stirred at room temperature for 18 h under nitrogen. The reaction mixture was quenched with ice. The organic layer was separated washed with brine, dried with anhydrous sodium sulfate and the solvent removed to get the crude product which was purified by flash chromatography to get benzyl octyl adipate (53-2, 650 mg, 21.3 %) as a colorless oil^H NMR (400 MHz, CDCh, ppm): δ 7.32-7.36 (m, 5H), 5.11 (s, 2H), 4.05 (t, 2H, J 6.8 Hz), 2.29-2.40 (m, 4H), 1.56-1.72 (m, 6H), 1.21-1.36 (m, 10H), 0.88 (t, 3H, J 6.8 Hz).Synthesis of octyl adipate(53-3 in scheme for Compound 53)To a solution of benzyl octyl adipate (53-2 10.0 g, 29 mmol) in ethyl acetate (100 mL) was added palladium hydroxide on carbon (20%) (1.5 g, 14 mmol) and the reaction mixture stirred at room temperature under hydrogen atmosphere for 18 h. The reaction mixture was filtered through a bed of celite, washed with ethyl acetate and dichloromethane and the solvent removed to get the crude product to get the product octyl adipate(53-3, 4.6 g, 49%), which was directly used in the next step. (4001H M NHMz,R CDCI3, ppm): δ 4.06 (t, 2H, J 6.8 Hz), 2.31-2.40 (m, 4H), 1.59-1.69 (m, 6H), 1.12-1.38 (m, 10H), 0.88 (t, 3H, J 6.8 Hz)Synthesis of 3-(heptadecan-9-yloxy)-3-oxopropanoic acid (53-5in scheme for Compound 53)A solution of 2, 2-dimethyl-l,3-dioxane-4, 6-dione (53-4, 4.0 g, 28 mmol) and heptadecan-9- 01 (51-14, 7.12 g, 28 mmol) in toluene (100 mL) was refluxed at 120 °C for 4 h. It was cooled and the solvent removed under vacuum to get the crude product, which was purified by flash chromatography using ethyl acetate and hexane to get pure 3-(heptadecan-9-yloxy)-3- oxopropanoic acid (53-5, 5.94 g, 62%) .[pH NMR (400 MHz, CDCI3, ppm): δ 4.93-4.99 (m, 1H), 3.42 (s, 2H), 1.54-1.56 (bm, 5H), 1.12-1.42 (m, 24H), 0.87 (t, 6H, J 6.6 Hz).Synthesis of 2-((2-((tert-butyldimethylsilyl)oxy)ethyl)amino)ethan-l-ol (53-6 in scheme for Compound 53)A solution of (2-bromoethyl)(tert-butyl)dimethylsilane (10.0 g, 42 mmol) and ethanolamine (51-9, 51.0 g, 50.5 mL, 840 mmol) in acetonitrile (130 mL) was heated to reflux at 90 °C for 3 h. It was cooled and poured into ice-water and extracted with ethyl acetate (2 X 100 mL). The organic layer was washed with brine, dried with anhydrous sodium sulfate, the solventremoved to get the crude product which was purified by flash chromatography to get 2-((2- ((tert-butyldimethylsilyl)oxy)ethyl)amino)ethan-l-ol (53-6, 3.78 g, 41%).1H NMR (400 MHz, CDCI3, ppm): δ 3.72 (t, 2H, J 5.4 Hz), 3.64 (t, 2H, J 5.0 Hz), 2.80 (t, 2H, J 5.2 Hz), 2.74 (t, 2H, J 5.2 Hz), 2.31 (bs, 2H), 0.90 (s, 9H), 0.07 (s, 6H).Synthesis of 2-((2-(benzyloxy)ethyl)(2-((tert-butyldimethylsilyl)oxy)ethyl)amino)ethan-l-ol (53-7 in scheme for Compound 53)To a solution of 2-((2-((tert-butyldimethylsilyl)oxy)ethyl)amino)ethan-l-ol (53-6, 3.5 g, 16 mmol) in acetonitrile (20 mL) was added ((2-bromoethoxy)methyl)benzene (5.15 g, 24 mmol) followed by potassium carbonate (6.6 g, 48 mmol) and potassium iodide (1.3 g, 8 mmol). The reaction mixture was then heated to reflux at 98 °C for 48 h. The solvent was removed and the crude compound purified by flash chromatography to get the pure product 2-((2-(benzyloxy)ethyl)(2-((tert-butyldimethylsilyl)oxy)ethyl)amino)ethan-l-ol (53-7, 3.87 g, 68 %).1H N (M40R0 MHz, CDCI3, ppm): δ 7.26-7.34 (m, 5H), 4.51 (s, 2H), 3.66 (t, 2H, J 6.0 Hz), 3.52-3.56 (m, 4H), 2.83 (t, 2H, J 5.8 Hz), 2.71-2.75 (m, 4H), 0.88 (s, 9H), 0.04 (s, 6H).Synthesis of 2-((2-(benzyloxy)ethyl)(2-((tert-butyldimethylsilyl)oxy)ethyl)amino)ethyl octyl adipate (53-8 in scheme for Compound 53)To a solution of octyl adipate (53-3, 1.5 g, 5.8 mmol) in dichloromethane (10 mL) at 0 °C was added oxalyl chloride (1.10 g, 0.75 mL, 8.7 mmol) followed by / V, / V-Dimethylformamide (0.05 mL) under nitrogen atmosphere. The reaction mixture was stirred at 0 °C for 2 h. Thesolvent was removed, and the crude reaction mixture dried in rotavapor for 2 h. Then the crude acid chloride was dissolved in dichloromethane (10 mL) and 2-((2-(benzyloxy) ethyl)(2- ((tert-butyldimethylsilyl)oxy)ethyl)amino)ethan-l-ol (53-7, 3.08 g, 8.7 mmol) was added at 0 °C followed by pyridine (0.92 g, 0.94 mL, 11.6 mmol). The reaction mixture was stirred at room temperature for 18 h under nitrogen. The reaction mixture was quenched with ice. The organic layer was separated washed with brine, dried with anhydrous sodium sulfate and the solvent removed to get the crude product which was purified by flash chromatography to get 2-((2-( benzy loxy)ethy l)(2-((tert- butyldimethylsilyl)oxy)ethyl)amino)ethyl octyl adipate (53-8 2.61 g, 76 %) as a colorless oil.XH NMR (400 MHz, CDCI3, ppm): δ 7.26-7.34 (m, 5H), 4.51 (s, 2H), 4.12 (t, 2H, J 5.6 Hz), 4.05 (t, 2H, J 6.8 Hz), 3.66 (t, 2H, J 6.4 Hz), 3.53 (t, 2H, J 6.8 Hz), 2.83-2.85 (m, 4H), 2.72 (t, 2H, J 5.2 Hz), 2.25-2.34 (m, 4H), 1.58-1.72 (m, 6H), 1.20-1.39 (m, 10H), 0.86-0.89 (m, 12H), 0.04 (s, 6H).Synthesis of 2-((2-(benzyloxy)ethyl)(2-hydroxyethyl)amino)ethyl octyl adipate (53-9 in scheme for Compound 53)To a solution 2-((2-(benzyloxy)ethyl)(2-((tert-butyldimethylsilyl)oxy)ethyl)amino)ethyl octyl adipate (53-8, 1.4 g, 2.5 mmol) in tetra hydrofuran (10 mL) at 0 °C, was added hydrofluoric acid - pyridine (2.5 mL, 70% HF - 30% Pyridine) and the reaction mixture stirred at room temperature for 18 h. The reaction was quenched with saturated sodium bi-carbonate solution, extracted with dichloromethane. The organic layer was washed with brine, dried with anhydrous sodium sulfate and the solvent removed to get the crude product which was used purified using silica gel column to get pure 2-((2-(benzyloxy)ethyl)(2- hydroxyethyl)amino)ethyl octyl adipate (53-9, 830 mg, 73%). (400 M1HHz, N CMDCRI3, ppm): δ 7.26-7.35 (m, 5H), 4.51 (s, 2H), 4.15 (t, 2H, J 5.6 Hz), 4.05 (t, 2H, J 6.8 Hz), 3.52-3.55 (m, 4H), 2.80-2.86 (m, 4H), 2.73 (t, 2H, J 5.2 Hz), 2.28-2.32 (m, 4H), 1.59-1.68 (m, 7H), 1.20- 1.39 (m, 10H), 0.88 (t, 3H, J 7.2 Hz).Synthesis of 2-((2-(benzyloxy)ethyl)(2-((3-(heptadecan-9-yloxy)-3-oxopropanoyl)oxy)ethyl) amino)ethyl octyl adipate 53-10 in scheme for Compound 53)Chemical Formula: C47H81NO9 Exact Mass: 803.59 Molecular Weight: 804.16To a solution of 3-(heptadecan-9-yloxy)-3-oxopropanoic acid (53-5, 290 mg, 0.85 mmol) in dichloromethane (10 mL) at 0 °C was added oxalyl chloride (161 mg, 0.1 mL, 1.27 mmol) followed by / V, / V-Dimethylformamide (0.05 mL) under nitrogen atmosphere. The reaction mixture was stirred at 0 °C for 2 h. The solvent was removed, and the crude reaction mixture dried in rotavapor for 2 h. Then the crude acid chloride was dissolved in dichloromethane (10 mL) and 2-((2-(benzyloxy)ethyl)(2-hydroxyethyl)amino)ethyl octyl adipate (53-9, 609 mg, 1.27 mmol) was added at 0 °C followed by pyridine (134 mg, 0.14 mL, 1.69 mmol). The reaction mixture was stirred at room temperature for 18 h under nitrogen. The reaction mixture was quenched with ice. The organic layer was separated washed with brine, dried with anhydrous sodium sulfate and the solvent removed to get the crude product which was purified by flash chromatography to get the pure product 2-((2- (benzyloxy)ethyl)(2-((3-(heptadecan-9-yloxy)-3-oxopropanoyl)oxy)ethyl)amino)ethyl octyl adipate (53-10, 200 mg, 29 %) as colorless oil. (4001H M NHMz,R CDCI3, ppm): δ 7.26-7.36 (m, 5H), 4.88-4.91 (m, 1H), 4.51 (s, 2H), 4.19 (t, 2H, J 6.4 Hz), 4.11 (t, 2H, J 6.2 Hz), 4.05 (t, 2H, J 6.8 Hz), 3.53 (t, 2H, J 5.8 Hz), 3.34 (s, 2H), 2.81-2.86 (m, 6H), 2.29-2.32 (m, 4H), 1.48- 1.65 (m, 9H), 1.20-1.39 (m, 35H), 0.85-0.89 (m, 9H).Synthesis 2-((2-((3-(heptadecan-9-yloxy)-3-oxopropanoyl)oxy)ethyl)(2- hydroxyethyl)amino) ethyl octyl adipate (Compound 53)To a solution of 2-((2-(benzyloxy)ethyl)(2-((3-(heptadecan-9-yloxy)-3- oxopropanoyl)oxy)ethyl) amino)ethyl octyl adipate (53-10, 200 mg, 0.25 mmol) in ethyl acetate (20 mL) at room temperature was added palladium hydroxide on carbon (20 %) (100 mg, 0.14 mmol) and the reaction mixture stirred under hydrogen atmosphere for 24 h. MS showed complete conversion. The reaction mixture was filtered through a bed of celite, washed with ethyl acetate, dichloromethane. The solvent was removed and the crude product purified by flash chromatography to get pure 2-((2-((3-(heptadecan-9-yloxy)-3- oxopropanoyl)oxy)ethyl)(2-hydroxyethyl)amino)ethyl octyl adipate (Compound 53, 100 mg, 56%) as colorless oil.1H ( N40M0R MHz, CD3OD, ppm): δ 4.88-4.91 (m, 1H), 4.20 (t, 2H, J 5.8 Hz), 4.13 (t, 2H, J 5.6 Hz), 4.04 (t, 2H, J 6.8 Hz), 3.51-3.53 (m, 2H), 3.37 (s, 2H), 2.72-2.84 (m, 8H), 2.29-2.33 (m, 4H), 1.51-1.65 (m 9H), 1.24-1.28 (m, 34H), 0.85-0.88 (m, 9H). APCI: m / z 714.6, HPLC-ELSD purity = >99%. HPLC-CAD purity = >97%. HPLC conditions:Column: Agela C18 column, 4.6 X 50 mm, 3 pm.Eluents: A: Acetonitrile with 0.1 % TFA, B: Water with 0.1 % TFA.Flow rate: 1 mL / min; Column Temperature: Room temperature (25 °C, not controlled)Gradient elution on HPLC:ELSD detector parameters: Evaporator temperature = 60C, Nebulizer Temperature = 60C, GasFlow Rate = 1.60 SLMUPLC conditions: Aquity C18 column, 3.0 X 150 mm, 1.7 pm.Eluents: A: Water with 0.1 % TFA, B: Acetonitrile with 0.1 % TFA.Flow rate: 0.5 mL / min; Column Temperature: 55 °C; Preheater Temperature: 55 °CGradient elution on UPLC:CAD detector: Evaporator temperature setting 35°CFormulation of LNPs with a messenger RNAThis is a general description of the use of an ionizable cationic lipid of the invention (molecular weight range approx. 500-1000) to formulate an LNP dispersion with a smallmedium size mRNA (for example 500-4000 nucleotides) An ionizable lipid of the invention, 1,2- distearoyl-sn-glycero-3-phosphocholine (DSPC), cholesterol, and 1,2-dimyristoyl-rac- glycero-3-methylpolyoxyethylene (DMG-PEG) are combined in a 40:15:43:2 molar ratio in anhydrous ethanol at a concentration 12.5 mM (1.85 mg / mL). mRNA was diluted in RNase- free 50 mM citrate buffer pH 4.0 to obtain a lipid:mRNA mass ratio of 10:1 (ionizable lipidmucleotide 3:1 molar ratio, often referred to as N:P ratio). The lipid solution in ethanol is then rapidly mixed in a 3:1 (aqueous to ethanol) volume ratio through a micromixer chip using a Nanoassemblr or Egnyte benchtop instrument (Precision Nanosystems) at a total flow rate (TFR) of 12 mL / min. This combination of concentrations and mixing ratio results in a 3:1 molar ratio of ionizable cationic lipid to mRNA phosphate groups and a total lipid to mRNA mass ratio of approximately 10:1. An alternative approach to achieve 3:1 molar ratio of ionizable cationic lipid to mRNA phosphate groups and a total lipid to mRNA mass ratio of approximately 20:1 uses 25 mM lipid stock (double the concentration in ethanol) with the same mRNA concentration at the start and the same mixing configuration. This composition may be preferably in some cases for more complete encapsulation. The mixed LNP dispersion in ethanol / water resulting from the encapsulation step is diluted 10-fold into 50 mM citrate buffer at pH 6 and subjected to tangential flow filtration (TFF) using a 300kmolecular weight cut-off membrane (mPES) until concentrated to the original volume. Subsequently, the citrate buffer is replaced with a buffer containing 10 mM Tris buffer at pH7.5 (measured at 20-25C), 80 mM sodium chloride, and 3% sucrose using diafiltration with 10 diavolumes. The LNP dispersion is concentrated to a volume that leads to no more than about 0.5 mg / mL mRNA concentration, as measured by Ribogreen (QUANT-IT kit, ThermoFisher) with TritonX to access the RNA for quantification. Then the LNP is filtered using a 0.2 micron PES syringe filter, aliquoted into vials, and frozen at l°C / min using a Corning® CoolCell® LX Cell Freezing Container until the samples reach -80°C. Samples are stored at -80°C and thawed on wet ice before analysis or use. The total RNA concentration, the percentage of input RNA recovered (% recovery), and the encapsulation efficiency (%encaps.) are determined using a Ribogreen assay, which is described elsewhere. The Z-avg diameter (nm) and polydispersity index (PDI) are measured using dynamic light scattering (Malvern Zetasizer) following a 1:100 dilution in phosphate buffered saline (PBS).Biophysical characterization of lipid nanoparticlesThe messenger RNA-containing LNP composition is characterized using analytical methods to determine the loading of messenger RNA, the percentage of messenger RNA that is encapsulated, and the size of the particles. The total amount of messenger RNA contained in the sample and the percentage of that messenger RNA that is encapsulated is determined using a fluorescence assay employing Ribogreen, a dye that becomes more emissive upon binding messenger RNA and the fluorescence relates quantitatively to the amount of RNA to which the dye binds. The total amount of messenger RNA is determined by disrupting the LNP with 1 wt% Triton-X 100 to expose the encapsulated messenger RNA, adding the dye, and comparing the emission intensity against a standard curve prepared using ribosomal RNA. The amount of unencapsulated messenger RNA is measured in a similar manner with the detergent disruption of the LNP is omitted. With the total amount of messenger RNA known and the amount of unencapsulated messenger RNA known, the percent encapsulated messenger RNA is calculated by the following formula:Percent Encapsulation (%) = ((RNATOTAL - RNAUNENCAPSULATED) / RNATOTAL) X 100 where RNATOTAL and RNAUNENCAPSULATED are, respectively, the concentrations of total messenger RNA and unencapsulated messenger RNA. The total messenger RNA content varies based on formulation, but generally fall in the range of 0.030-0.200 mg / mL. The size of LNP is measured using dynamic light scattering of a sample diluted 1:100 in PBS buffer. pKa' ProtocolIn a black 96-well plate, solutions of mRNA-LNP (final assay concentration 2 pg / mL total RNA) in a series of buffers ranging from pH 4 to 9.5 are prepared. Buffers between pH 4 to7.6 are prepared from disodium phosphate and citric acid. Buffers from 7.8 to 9.5 are prepared by titrating Tris buffer with 10 N sodium hydroxide. To each well is added 6-(p- Toluidino)-2-naphthalenesulfonic acid sodium salt (TNS) in water to a final assay concentration of 6 pM. The fluorescence is read on a plate reader at 25°C with an excitationsetting of 321 nm and an emission setting of 445 nm. The intensity values are plotted as a function of pH using GraphPad Prism and are fit with a sigmoidal dose response curve. The apparent pKa (pKa') is determined as the EC50 of this curve where half of the ionizable amines are expected to be protonated.Characterization of the in vitro cellular potency of LNPsThe ability of an LNP to transfect cultured cells with a TNA, e.g., mRNA, is characterized using an in vitro assay based on the percentage of cells expressing the protein of interest. Specifically, 1 million BHK-21 cells are co-incubated with mRNA-LNP of varying concentration (highest 6.25 ng per well, lowest 0.39 ng per well) in 0.3 mL of media for 17- 19 hours at 37°C with 5% CO2.Characterization of the in vivo potency of lipid nanoparticlesThe in vivo potency of messenger RNA-LNPs may be evaluated using two related, but distinct, approaches. Firstly, to quantify the location, relative amount, and the duration of protein expression, LNPs formulated with a messenger RNA expressing luciferase is injected into mice, for example intramuscularly in the hind leg muscles. At defined time points, such as daily, the mice are administered luciferin, and the bioluminescence is imaged and quantified. Secondly, the ability of messenger RNA-LNPs to act as a vaccine is evaluated by measuring the antibody- and cell based immune response following a prime-boost vaccination schedule. For example, a priming vaccination given on Day 0 via intramuscular injection (i.m.) is followed 21 days later with a boosting vaccination. After an additional 21 days (Day 42 of the experiment), the mice are sacrificed and relevant tissues, such as serum, the spleen, or lymph nodes are collected for further analysis. Serum is analyzed using an ELISA to determine the antigen- specific antibody response. Splenocytes are analyzed for antigen- specific cytokine production, for example by using flow cytometry. Taken together, these in vivo assays can demonstrate that the messenger RNA-LNP is able to induce protein expression of the antigen of interest that initiates a productive antigen- specific immune response required for effective vaccination.All patents and other publications; including literature references, issued patents, published patent applications, and co-pending patent applications; cited throughout this application are expressly incorporated herein by reference for the purpose of describing and disclosing, for example, the methodologies described in such publications that might be used in connection with the technology described herein. These publications are provided solely for their disclosure prior to the filing date of the present application. Nothing in this regard should be construed as an admission that the inventors are not entitled to antedate such disclosure by virtue of prior disclosure or for any other reason. All statements as to the date or representation as to the contents of these documents is based on the information available to the applicants and does not constitute any admission as to the correctness of the dates or contents of these documents. The description of embodiments of the disclosure is not intended to be exhaustive or to limit the disclosure to the precise formdisclosed. While specific embodiments of, and examples for, the disclosure are described herein for illustrative purposes, various equivalent modifications are possible within the scope of the disclosure, as those skilled in the relevant art will recognize. For example, while method steps or functions are presented in a given order, alternative embodiments may perform functions in a different order, or functions may be performed substantially concurrently. The teachings of the disclosure provided herein can be applied to other procedures or methods as appropriate. The various embodiments described herein can be combined to provide further embodiments. Aspects of the disclosure can be modified, if necessary, to employ the compositions, functions and concepts of the above references and application to provide yet further embodiments of the disclosure. Moreover, due to biological functional equivalency considerations, some changes can be made in protein structure without affecting the biological or chemical action in kind or amount. These and other changes can be made to the disclosure in light of the detailed description. All such modifications are intended to be included within the scope of the appended claims. Specific elements of any of the foregoing embodiments can be combined or substituted for elements in other embodiments. Furthermore, while advantages associated with certain embodiments of the disclosure have been described in the context of these embodiments, other embodiments may also exhibit such advantages, and not all embodiments need necessarily exhibit such advantages to fall within the scope of the disclosure. The technology described herein is further illustrated by the following examples which in no way should be construed as being further limiting. It should be understood that this disclosure is not limited in any manner to the particular methodology, protocols, and reagents, etc., described herein and as such can vary. The terminology used herein is for the purpose of describing particular embodiments only and is not intended to limit the scope of the present disclosure, which is defined solely by the claims.LNP formulation with astaxanthin prodrug in LNPs.The following description is an example of formulating an LNP dispersion with a smallmedium size mRNA (for example 500-4000 nucleotides) and a fifth lipid component additive. An ionizable lipid (IL) of the invention and / or known lipid (e.g., DLin-MC3-DMA ("MC3")) or LP01, 1,2- distearoyl-sn-glycero-3-phosphocholine (DSPC), cholesterol (chol), astaxanthin amino prodrug (AAP, e.g. compound 90 or 89, which are ionizable) and 1,2- dimyristoyl-rac-glycero-3-methylpolyoxyethylene (DMG-PEG) are combined in ratios that produces a nanoparticle dispersion having suitable size and polydispersity, exhibiting RNA encapsulation and activity in a biological assay.DLin-MC3-DMA ("MC3")1,2- distearoyl-sn-glycero-3-phosphocholine ("DSPC")dioleyl-phosphatidylethanolamine ("DOPE")A particular example is IL:DSPC:chol:AAP:DMG-PEG, having molar ratio of 45:12:39:2:2, in anhydrous ethanol at a concentration 12.5 mM (1.85 mg / mL). Modified mRNA encoding for firefly Luciferase using 5-methoxyuridine substitutions for all uridines, was diluted in RNase- free 50 mM acetate buffer pH 4.0 to obtain a lipid:mRNA mass ratio of 10:1 (ionizable lipidmucleotide 4.5:1 molar ratio, often referred to as N:P ratio). The lipid solution in ethanol is then rapidly mixed in a 3:1 (aqueous to ethanol) volume ratio through a micromixer chip using a Nanoassemblr, Egnyte benchtop instrument (Precision Nanosystems) or similar mixer setup at a total flow rate (TFR) of 12 mL / min. This combination of concentrations and mixing ratio results in a 3:1 to 4.5:1 molar ratio of ionizable lipid to mRNA phosphate groups and a total lipid to mRNA mass ratio of approximately 10:1 - 15:1. The mixed LNP dispersion in ethanol / water resulting from the encapsulation step is diluted 10-fold into 50 mM citrate buffer at pH 6 and subjected to dialysis against phosphate buffered saline (PBS). Alternatively, tangential flow filtration (TFF) using a 300kDa molecular weight cut-off membrane (mPES) is applied until concentrated to the original volume. Subsequent to TFF, the citrate buffer is replaced with a buffer containing 10 mM Tris buffer at pH 7.5 (measured at around 20°C), 80 mM sodium chloride, and 3% sucrose using diafiltration with 10 diavolumes. The LNP dispersion is concentrated to a volume that leads to no more than about 0.5 mg / mL mRNA concentration, as measured by Ribogreen (QUANT-IT kit, ThermoFisher) with TritonX to rupture LNPs and access the encapsulated RNA for quantification. Then the LNP is filtered using a 0.2 micron PES syringe filter, aliquoted into vials, and frozen at l°C / min using a Corning® CoolCell® LX Cell Freezing Container until the samples reach -80°C. Samples are stored at -80°C and thawed on wet ice before analysis or use. The total RNA concentration, the percentage of input RNA recovered(% recovery), and the encapsulation efficiency (%encapsulation) are determined using a Ribogreen assay. The Z-avg diameter (nm) and polydispersity index (PDI) are measured using dynamic light scattering (Malvern Zetasizer) following a 1:100 dilution in phosphate buffered saline (PBS). LNPs are then stored refrigerated and in the dark.Biophysical characterization of lipid nanoparticlesThe messenger RNA-containing LNP composition is characterized using analytical methods to determine the loading of messenger RNA, the percentage of messenger RNA that is encapsulated, and the size of the particles. The total amount of messenger RNA contained in the sample and the percentage of that messenger RNA that is encapsulated is determined using a fluorescence assay employing Ribogreen, a dye that becomes more emissive upon binding messenger RNA. The total amount of messenger RNA is determined by disrupting the LNP with 1 wt% Triton-X 100 to expose the encapsulated messenger RNA, adding the dye, and comparing the emission intensity against a standard curve prepared using ribosomal RNA. The amount of unencapsulated messenger RNA is measured in a similar manner with the detergent disruption of the LNP is omitted. With the total amount of messenger RNA known and the amount of unencapsulated messenger RNA known, the percent encapsulated messenger RNA is calculated thus:Percent Encapsulation (%) = ((RNATOTAL - RNAUNENCAPSULATED) / RNATOTAL) X 100 where RNATOTAL and RNAUNENCAPSULATED are, respectively, the concentrations of total messenger RNA and unencapsulated messenger RNA. The total messenger RNA content varies based on formulation, but generally fall in the range of 0.030-0.200 mg / mL. The size distribution parameters of LNP are measured using dynamic light scattering of a sample diluted 1:100 in PBS buffer.Fluorescence imaging and spectroscopy can be used to identify and quantify xanthophyll components in the LNP. The fluorescence is read on a plate reader at 25°C with an excitation setting of 450 nm and an emission setting of 560 nm.Table 1. LNP compositions used to generate data presented in Figures 1-7 and 9-11. RNA content and encapsulation was generated with RiboGreen assay.Table 2. LNP compositions used to generate data presented in Figures 12 and 13. RNA content and encapsulation was generated with RiboGreen assayTable 3. LNP compositions used to generate data presented in Figure 14.Expression of Luciferase from modified mRNA by in vitro cell transfection with LNPs.LNPs from the preparations entrapping the Firefly Luciferase mRNA with uridine modifications were dosed into wells containing a single cell line, either HEK293 or Huh7, at doses of 125, 250 and 500 ng of mRNA, in addition to a PBS blank dose denoted as 0 ng mRNA. After 6 hours, the luminescence from the expressed Luciferase is tested after dosing of Luciferin dye, a substrate for the enzyme which is luminescent, and the signal is proportional to the enzyme activity. Viability of cells is measured by Promega OneGlo + Tox luciferase detection kit.A set of experiments was done to test the potential of LNPs compositions with proprietary lipids designed by and synthesized.The choice of helper lipid, specifically the phospholipids, was aimed to compare DSPC (used in most formulations with RNA, including ONPATTRO and the mRNA vaccines for Covidl9) as well as DOPC and DOPE.Additionally, evaluation of MC3 (D-Lin-MC3-DIVIA) as a potent and ionizable cationic lipid was done. MC3 is used for the effective delivery of RNAs in vivo, although it is known to be reactogenic induce immuno-inflammatory responses (either TH1 or TH2).In the studies, all the ionizable lipid formulations tested containing MC3 with or without AAP modifiers, Compound (89) and Compound (90) (Figure 1A) or with compositions and structures of Axelyf ionizable lipids as per Table 1 (Figure IB) yielded particles withing expected optimal range. Additionally, the polydispersity (PDI) measured using Stunner (rotating angle dynamic light scattering, Unchained Labs) Protocol was also within ranges expected from this type of lipid nanoparticles (Figure 2A and Figure 2B). Importantly, most were able to yield an RNA encapsulation efficiency >80%, which is a lower threshold for what is considered good encapsulation of mRNA (Table 1). The presence of astaxanthin compounds did not dramatically affect the measured properties of the particles, although Compound (90) addition resulted in overall larger particles. Notably, assessment of size (Figure 3A) and polydispersity (Figure 3B) of LNPs with and without AAP modifier component on day 14 after formulation showed that particles retained their overall characteristic in storage.All MC3-based LNPs showed similar pKa' values in line with literature reports (value range of 6.3-6.6).Functional expression of luciferase was measured by luminescence, a direct enzymatic activity readout, in human embryonic kidney 293T cells (HEK293T) and in hepatic cell line Huh-7 (both epithelial-like), with increasing mRNA payload (125-250-500ng). LNP formulations containingboth standard and reduced molar ratio of MC3, with or without Axelyf AAP proprietary ionizable lipids induced dose-dependent luciferase expression luciferase expression, with or without AAP modifier components showed satisfactory in HEK293T cells (Figure 4A and 4B, respectively) with notably lower expression for Compound (52) in formulations 7 and 9. Dose dependent luciferase expression with (Figure 5A) or without (Figure 5B) AAP modifier was also observed in Huh7. In this cell type, formulation with Compound (52), i.e., formulations 7 and 9 showed the highest luciferase expression, suggesting possible in vivo differential tropism.All the LNPs formulation tested showed acceptable minimal toxicity at the tested RNA concentrations based on cell viability measurements in HEK293T cells (Figure 6A and Figure 6B.) and in Huh7 cells (Figure 7A and 7B), regardless of the presence of AAP modifiers or lack thereof.After a one-month storage at +4°C, the size and polydispersity of particles showed maintained consistency, with the exception of MC3 Compound (90 containing LNPs, which, already larger than other compositions, showed an increase in size over the observation period / Encapsulation efficiency of particles did not appear to change significantly on storage.After one month, dose-dependent expression of luciferase was consistent with earlier experiments in all MC3-based LNPs, with only minor differences among the various formulations (in both cell types). Notably, expression of luciferase in Huh-7 induced by MC3 LNPs, including the ones with the astaxanthin derivatives, was higher than in 293T cells.Conversely, formulation Compound (30) using DSPC yielded a strikingly high expression of luciferase in HEK293T cells as compared with all other formulations, suggesting good mRNA stability, and formulation Compound (52) DOPE drove expression in Huh7, whereas other formulations had weaker activity. After one month at +4°C, different formulations retained different tropism.In order to further evaluate the contribution of astaxanthin derivative Compound (89), a further set of experiments was performed comparing MC3 LNPs containing increasing molar percentages (2%, 4-%, 8%) of Compound (89), with unmodified 2% astaxanthin as control. Visible light assessment of the color of formulations with increasing concentrations of Compound (89)-and astaxanthin showed a progressively increasing red color of Compound (89) formulations.Fluorescence spectral scan revealed emission of Compound (89) and Astaxanthin 2% in EtOH around 575 nm with 480 nm excitation, but little fluorescence of the lipid nanoparticles containing these compounds (Figure 9).All of the ionizable lipid formulations tested produced measurable particles, with the size (Figure 11, left) and PDI (Figure 11, right) within expected from this type of lipid nanoparticles. All LNPs were able to yield an RNA encapsulation efficiency >90% (Table 1). Astaxanthincompounds did not dramatically affect the measured size or encapsulation properties of the particles.Transfection potential and cytotoxicity of all LNPs were measured once again measured in HEK293T and Huh7. All Compound (89) formulations showed slightly higher expression of luciferase in both cell lines in compared to MC3 control, with 4% Compound (89) showing also better viability. Astaxanthin inclusion in LNPs did not provide the same increase in luciferase expression over control that was seen with Compound (89) in either cell line (Figure 12). In HEK293T and Huh7 cells there appeared to be minimal toxicity of LNPs with Compound (89) at tested RNA concentrations based on cell viability measurements when compared to MC3 LNPs alone. However, formulation with 8% Compound (89) showed some toxicity in both cell lines at the highest concentration (500ng; Figure 13).LNPs compositions with ionisable lipids Cpd 51 or Cpd 53 were made and compared to LNP with MC3 and LP-01 which are widely used in the literature.LNP containing Cpd 51 or Cpd 53 (with either DOPE, DOPC, or DSPC) had Z-average values in line with control LNPs (Figure 13A). PDI values were lower than MC3 LNPs, but in line withLP-01 LNPs (Figure 13B).LNPs containing Cpd 51 or Cpd 53 were equipotent to LP-01 in terms of in vitro expression of Luciferase in HEK293T and Huh7 cells as a result of transfection of different amount of modified mRNA.

Claims

CLAIMSWe claim:

1. A lipid having a structural formula selected from the group consisting of:and (4); or a pharmaceutically acceptable salt or cocrystal thereof, wherein, for each occurrence:* denotes a chiral center;R1for each occurrence, is independently selected from: -H, optionally substituted C1-C6 alkyl, optionally substituted C2-C& alkenyl, -C(=O)CH2N(R72)2,-P(O)(OR17)(OR18), -P(O)(OR17)(NR19), or -P(O)(NR19)(NR20);R3for each occurrence, is independently selected from: -H, optionally substituted C1-C6 alkyl, -[CI-hJi-eOH, optionally substituted C2-C6 alkenyl, -P(O)(OR17)(OR18), - P(O)(OR17)(NR19), -P(O)(NR19)(NR20) or R4;R2is selected from: is absent (i.e., a lone pair on N), -H or optionally substituted C1-C6 alkyl; provided that when R2is -H or optionally substituted C1-C6 alkyl, the nitrogen atom, to which R1, R2, and R3are all bonded to, is protonated;R4for each occurrence, is independently selected from:orR55for each occurrence, is independently selected from:A1, A3, A4, A5, A6, A7, A8, A12and A13for each occurrence, are each independently selected from: a bond, -O-, -C(=O)-, -C(=S)-, -OC(=O)-, -C(=O)O-, -OC(=O)O-, -OC(=S)-, -C(=S)O-, - OC(=S)O-,-S-C(=O)-, -C(=O)-S-, -S-S-, -S(OH)-, -S(=O)-, -S(=O)2-, -S(OH)2-, -C(=O)N(R14)-, - N(R14)C(=O)-, -N(R14)C(=O)N(R14)-, -O-C(=O)C(R14)2C(=O)O-, -C(=O)O-C(R14)2C(=O)O-, -O- C(=O)C(R14)2-O-C(=O)-, -O-C(R14)2C(=O)O-, -O-C(=O)C(R14)2C-O-, -O-C(R14)2-O-C(=O)-, - C(=O)-O-C(R14)2C-O-, -O-C(=O)-O-R41-O-C(=O)O-, -0-C(=0)-[CH2]O-4-C(=0)-0-, -P(OH)-, - P(OH)(R14)-, -P(=O)(OH)-, -P(O R14)2-, -P(O R14)- or -P(=O)(O R14)-;A2for each occurrence, is independently selected from: a bond, -O-, -OC(=O)-, -C(=O)O-, - OC(=O)O-, -S-C(=O)-, -C(=O)-S-, -S-S-, -C(=O)N(R14)-, -N(R14)C(=O)-, -N(R14)C(=O)N(R14)-, -O-C(=O)C(R14)2C(=O)O-, -C(=O)O-C(R14)2C(=O)O-, -O-C(=O)C(R14)2-O-C(=O)-, -O- C(R14)2C(=O)O-, -O-C(=O)C(R14)2C-O-, -O-C(R14)2-O-C(=O)-, -C(=O)-O-C(R14)2C-O- or -O- C(=O)-O-R41-O-C(=O)O-;R6for each occurrence, is independently selected from: a bond, -R58-N(R42)-R58-, -R58- CH(R42)-R58-, Ci-Ci2alkylene or C2-Ci2alkenylene;R7for each occurrence, is independently selected from: a bond, optionally substituted Ci- Ci8 alkylene, or optionally substituted C2-Cis alkenylene;R8and R9for each occurrence, are each independently selected from:R42;R12for each occurrence, is independently selected from: -H, optionally substituted Ci-Cie alkyl or optionally substituted C2-C16alkenyl;R13for each occurrence, is independently selected from: optionally substituted C1-C16 alkyl or optionally substituted C2-C16alkenyl;R14, for each occurrence, is independently selected from: -H, optionally substituted C1-C10 alkyl, optionally substituted C1-C10 alkenyl or optionally substituted C1-C10 alkynyl;R15for each occurrence, is independently selected from: a bond, -R41-N(R42)-R58-, -R41- CH(R42)-R58-, or C1-C6 alkylene;R16for each occurrence, is independently selected from: a bond or -[CH2]k-;R17and R18for each occurrence, are each independently selected from: -H, C1-C5 alkyl, an alkali metal cation, an alkaline earth metal cation, ammonium cation, methyl ammonium cation, or a pharmaceutically acceptable base;R19and R20for each occurrence, are each independently selected from: -OH, formyl, acetyl, pivaloyl, -NH2, -NH(CH3), -NH(CH2CH3), -N(CH3)2, -NHC(=O)H, -NHC(=O)CH3or C3-C5- alkyl;R29, R30, R31, R32, R33and R34for each occurrence, are each independently selected from: - CH2-, -NH-, -S-, or -O-;R35for each occurrence, is independently selected from: -OH or C1-C6 acyl; optionally substituted C1-C6 alkyl, C1-C6 alkyl ester, C1-C6 alkyl ether or C1-C6 alkyl carbonate;R36and R37are each independently selected from: -H, -C(=O)C1-C6 alkyl, -C(=O)C2-C6alkenyl, -C(=O)-O-C1-C6 alkyl, or -C(=O)-O-C2-C6alkenyl, C1-C6 alkyl, or C2-C6alkenyl;R38for each occurrence, is independently selected from: -H, C1-C24 alkylene or C1-C24 alkenylene;R41for each occurrence, is independently selected from: a bond or C1-C5 alkylene;R42for each occurrence, is independently selected from: C6-C24 alkyl, C6-C24 alkyl carbonate, C6-C24 alkyl ether, C6-C24 alkyl ester, -R58-A1-R59or -R58-A1-R59-A2-R60, wherein R42has 6-24 total carbon atoms;R58R59, and R60for each occurrence, are each independently selected from: a bond, -O-, - O-C(=O)-, -OC(=O)O-, Ci-C24alkylene or C2-C24alkenylene;R62, R63, R64, R67, R68, R69, R74, R75, R76, R77, R108, R109, R110and R111for each occurrence, are each independently selected from: a bond, cycloalkylene , C1-C24 alkylene, C1-C24 alkenylene, -R114-C3-C6-cycloalkylene-R115-, -R114-CH=CH-CH2-CH=CH-R115- , -R114- CH=CH-CH2-cPr-R115-, -R114-cPr-CH2-CH=CH-R115-, -R114-cPr-CH2-cPr-R115-, -R114-CH=CH- CD2-CH=CH-R115-, -R114-CH=CH-CD2-cPr-R115-, -R114-cPr-CD2-CH=CH-R115- or -R114-cPr- CD2-cPr-R115-; where -cPr- is cyclopropylene (cyclopropane-1,2 diyl); each double bond or cPr has a cis configuration; and D denotes deuterium (2H);R114and R115for each occurrence, are each independently selected from: a bond, C1-C10 alkylene, C1-C10 alkenylene C1-C10 alkynylene;R61, R73and R107for each occurrence, are each independently selected from:R116for each occurrence, is independently selected from: -H, -F, -CF3, -Cl, -Br, -I, -CH3, - CH2R14, -CHR142, -CR143, -OH, -OR14, -NH2, -NHR14, -N(R14)2, -SH, -SR14, -S(=O)H, - S(=O)R14, -S(=O)2H, -S(=O)2R14, -PH2, -PHR14, -PR142, -P(OH)2, -POHR14, -PR142, -P(OR14)2, -P(O)H2, -P(O)HR14, -P(O)R142, -P(O)(OH)2, -P(O)(OR14)OH, -P(O)(OR14)2, -SiH3, -SiH2R14, - SiHR142or-SiR143;R65, R70, R78, R105and R112for each occurrence, are each independently selected from: a bond, -CH=CH-CH2-CH=CH- , C3-C6-cycloakylene, -CH=CH-CH2-cPr-, -cPr-CH2-CH=CH-, - cPr-CH2-cPr-, -CH=CH-CD2-CH=CH- , -CH=CH-CD2-cPr-, -cPr-CD2-CH=CH- -or -cPr-CD2- cPr-, where -cPr- is cyclopropylene, each double bond or cPr has a cis configuration and D denotes deuterium (2H);R66, R71, R79, R106and R113for each occurrence, are each independently selected from: -H, linear or branched C1-C24 alkyl, C1-C24 alkenyl, or C1-C24 alkynyl, C5-C2o-spirocycloalkyl, C3-Ci8-cycloalkyl, C3-Ci8-heterocycloalkyl, optionally substituted aryl, optionally substituted heteroaryl, -R26-CH=CH-CH2-CH=CH-R28, -R26-CH=CH-CH2-cPr-R28, -R26-cPr- CH2-CH=CH-R28, -R26-cPr-CH2-cPr-R28, -R26-CH=CH-CD2-CH=CH-R28, -R26-CH=CH-CD2-cPr- R28, -R26-cPr-cPr-R28, -R26-cPr-cBu-R28, -R26-cBu-cBu-R28, -R26-cBu-cPr-R28, -R26-cHx-cBu- R28, -R26-cHx-cBu-R28, -R26-cHx-cBu-R28, -R26-cPr-cHx-R28, -R26-cBu-cHxR28, -R26-cPr-R26- cPr-R28, -R26-cPr-R26-cBu-R28, -R26-cBu-R26-cBu-R28, -R26-cBu-R26-cPr-R28, -R26-cHx-R26- cBu-R28, -R26-cHx-R26-cBu-R28, -R26-cHx-R26-cBu-R28, -R26-cPr-R26-cHx-R28, -R26-cBu-R26- cHxR28, -R26-c-CD2-cPr-R28, -R26-cPr-CD2-cPr-R28, -R26-cPr-CD2-cPr-R28, -R26-cPr-CD2- CH=CH-R28, -R26-cPr-CD2-cPr-R28or -R26-C3-Ce cycloakylene-R26-C3-Ce cycloakylene-R28; wherein, R26for each occurrence, is independently selected from: a bond, C1-C10 alkylene, C1-C10 alkenylene or C1-C10 alkynylene; and R28for each occurrence, is independently selected from: -H, C1-C10 alkyl, C1-C10 alkenyl or C1-C10 alkynyl; -cPr- is cyclopropylene (cyclopropane-1,2 diyl); -cBu- is cyclobutylene; -cHx- is cyclohexylene; each double bond or -cPr- has a cis configuration; and D denotes deuterium (2H);R72for each occurrence, is independently selected from; -H, optionally substituted C1-C6 alkyl, -[CH2]I-6OH or optionally substituted C2-C6alkenyl;R96for each occurrence, is independently selected from: -OH, -O-C1-C4 alkyl, -O-C(=O)-O- C1-C4 alkyl or -O-C(=O)-C1-C5 alkyl;R118and R119are independently selected from R1or R55;n is an integer selected from: 1, 2, 3, 4, 5 or 6; and k is an integer selected from: 1, 2, 3, 4, 5 or 6.

2. The lipid of claim 1, comprising one or more cycloalkylene independently selected from:

3. The lipid of claim 1, wherein, -R62-A3-R63-A4-R64-R65-R66, -R62-A3-R63-A4-R64-R73-R74-A7- R75-R65-R66, -R62-A3-R63-A4-R64-R73-R76-A8-R77-R78-R79, -R67-A5-R68-A6-R69-R70-R71, -R67-A5- R68-A6-R69-R107-R108-A12-R109-R105-R106, and -R67-A5-R68-A6-R69-R107-R110-A13-R111-R112-R113, for each occurrence, independently has: f) a total number of carbon atoms selected from: 10-24, 12-22, 14-22, 16-20, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23 or 24 carbon atoms; g) a longest linear chain with a total number of carbon atoms that is selected from: 10- 24, 12-22, 14-22, 16-20, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23 or 24 carbon atoms; or h) a longest linear chain with a total number of atoms that is selected from: 10-24, 12-22, 14-22, 16-20, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23 or 24 atoms.

4. The lipid of claim 1, comprising two R55variables selected from: a) -R62-A3-R63-A4-R64-R73-R74-A7-R75-R65-R66and -R62-A3-R63-A4-R64-R73-R76-A8-R77-R78-R79, b) -R74-A7-R75-R65-R66and -R76-A8-R77-R78-R79, c) -R62-A3-R63-A4-R64-R65-R66and -R67-A5-R68-A6-R69-R70-R71, d) -R67_A5_R68_A6_R69_R1O7_R1O8_A12_R1O9_R1O5_R1O6a n d_R67_A5_R68_A6_R69_R107_R110_A13_Rlll_R112-R113, or e) -R1O8.A12.R1O9.R1O5.R1O6a n d_R110_A13_Rlll_R112_Rll35. The lipid of claim 1, wherein R55, for each occurrence, is independently selected from:d) -R67-A5-R68-A6-R69-R70-R71, e) -R67-A5-R68-A6-R69-R107-R108-A12-R109-R105-R106f) _R67_A5_R68_A6_R69_RIO7_RIIO_AI3-RIII_RII2_RII3;g) -A3-R63-A4-R64-R73-R74-A7-R75-R65-R66, h) -A3-R63-A4-R64-R73-R76-A8-R77-R78-R79, i) -A3-R63-A4-R64-R65-R66, j) -A5-R68-A6-R69-R70-R71, k) -A5-R68-A6-R69-R107-R108-A12-R109-R105-R106l) -A5-R68-A6-R69-R107-R110-A13-R111-R112-R113, m) -R74-A7-R75-R65-R66, n) -R76-A8-R77-R78-R79, o) _R108_A12_R109_R105_R106_P)Riio_Ai3-Riii_Rii2_Rii3q) -R74-R66orr) -R76-R79.

6. The lipid of claim 1, wherein R55, for each occurrence, is independently selected from:-OC(=O)C(R42)[CH2]nC(=O)O-R41-CH(C4-Ci2 alkyl)2, -R41-N(R42)([CH2]nC(=O)O-R41-CH(C4-Ci2alkyl)2, -OC(=O)C(R43)[CH2]nC(=O)O-R41-CH(C4-Ci2alkyl)2, -R41-N(R43)([CH2]nC(=O)O-R41- CH(C4-CI2alkyl)2, -R41-C(R42)[CH2]nC(=O)O-R41-CH(C4-Ci2alkyl)2or -R41-CH(R43)(R44), - R41-N(R43)(R44); wherein,R43for each occurrence, is independently selected from: -C(=O)O-C6-C24 alkyl, -OC(=O)-Ce- C24alkyl, -OC(=O)O-C6-C24alkyl, -C(=O)O-C6-C24 alkenyl, -O-C(=O)-C6-C24 alkenyl, - OC(=O)O-C6-C24alkenyl-, -C2-C22alkylene-O-C2-C22alkyl, -C2-C22alkenylene-O-C2-C22alkyl, -C2-C22alkylene-O-C2-C22alkenyl, -Ci-C22alkylene-C(=O)O-Ci-C22alkyl, -Ci-C22alkylene-O-C(=O)O-Ci-C22alkyl, -Ci-C22alkylene-O-C(=O)-Ci-C22alkyl, -C2-C22alkenylene-C(=O)O-Ci-C22alkyl, -C2-C22alkenylene-O-C(=O)-Ci-C22alkyl, -C2-C22alkenylene-O-C(=O)-Ci-C22alkyl, -Ci-C22alkylene-C(=O)O-C2-C22alkenyl, -Ci-C22alkylene-O-C(=O)O-C2-C22alkenyl, -Ci-C22alkylene-O-C(=O)-C2-C22alkenyl, -C2-C22alkenylene-C(=O)O-C2-C22alkenyl, -C2-C22alkenylene-O-C(=O)O-C2-C22alkenyl, -C2-C22alkenylene-O-C(=O)-C2-C22alkenyl; wherein R43has 6-24 total carbon atoms;R44for each occurrence, is independently selected from: -R45-C(=O)O-R45-CH(C6-CI2al kyl)2, -R45- OC(=O) - R45-CH(C6-C12 alkyl)2, -R45-OC(=O)O-R45-CH(C6-C12 alkyl)2, R45-C(=O)O-R45- CH(C6-C12 alkenyl)2, -R45- OC(=O) - R45-CH(C6-C12 alkenyl)2, -R45-OC(=O)O- R45-CH(C6-CI2alkenyl)2; wherein R44has 6-24 total carbon atoms and wherein R45is, for each occurrence, independently selected from: a bond, C1-C6 alkylene or C1-C6 alkenylene.

7. The lipid of claim 1, wherein R55, for each occurrence, is independently:wherein,A3and A5for each occurrence, are each independently selected from: -O-, -OC(=O)-, - C(=O)O- or -OC(=O)O-;R16for each occurrence, is independently selected from: a bond or — [CH2]k-; k is an integer selected from: 1, 2, 3, 4, 5 or 6; rence, are each independently selected from:h occurrence, are each independently selected from: C1-C12 alkylene or C1-C12 alkenylene;R66, R71and R79for each occurrence, are each independently selected from: C1-C12 alkyl or C1-C12 alkenyl.

8. The lipid of claim 7, wherein, R62, R63, R66and R74, together, have 10-24 total carbon atoms; R62, R63, R76and R79, together, have 10-24 total carbon atoms; and R67, R68and R71, together, have 10-24 total carbon atoms.

9. The lipid of claim 7, wherein, -R62-A3-R63-R73-R74-R66, -R62-A3-R63-R73-R76-R79, and -R67- A5-R68-R71, independently has: i) a total number of carbon atoms selected from: 10-24, 12-22, 14-22, 16-20, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23 or 24 carbon atoms; j) a longest linear chain with a total number of carbon atoms that is selected from: 10- 24, 12-22, 14-22, 16-20, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23 or 24 carbon atoms; or k) a longest linear chain with a total number of atoms that is selected from: 10-24, 12-22, 14-22, 16-20, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23 or 24 atoms.

10. The lipid of claim 1, wherein R55, for each occurrence, is independently:wherein:A3and A5for each occurrence, are each independently selected from: -O-, -OC(=O)-, -C(=O)O- or -OC(=O)O-;R16for each occurrence, is independently selected from: a bond or — [CH2]k-; k is an integer selected from: 1, 2, 3, 4, 5 or 6; each independently selected from:each occurrence, are each independently selected from: C1-C12 alkylene or C1-C12 alkenylene;R66, R79, R106and R113for each occurrence, are each independently selected from: C1-C12 alkyl or C1-C12 alkenyl; and wherein: R62, R63, R66and R74, together, have 10-24 total carbon atoms; R62, R63, R76and R79, together, have 10-24 total carbon atoms; and R67, R68, R108and R106, together, have 10-24 total carbon atoms; and, R67, R68, R110and R113, together, have 10-24 total carbon atoms.

11. The lipid of claim 7, wherein, -R62-A3-R63-R73-R74-R66, -R62-A3-R63-R73-R76-R79, -R67-A5-R68- R1O7_R1O8_R1O6a n d_R67_A5_R68_RIO7_RIIO_RII3 independently have: a) a total number of carbon atoms selected from: 10-24, 12-22, 14-22, 16-20, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23 or 24 carbon atoms;b) a longest linear chain with a total number of carbon atoms that is selected from: 10- 24, 12-22, 14-22, 16-20, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23 or 24 carbon atoms; or c) a longest linear chain with a total number of atoms that is selected from: 10-24, 12-22, 14-22, 16-20, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23 or 24 atoms.

12. The lipid of claim 1, wherein R55, for each occurrence, is independently:wherein:A3and A5for each occurrence, are each independently selected from: -O-, -OC(=O)-, - C(=O)O- or -OC(=O)O-;R16for each occurrence, is independently selected from: a bond or — [CH2]k-; k is an integer selected from: 1, 2, 3, 4, 5 or 6;R61, R73and R107for each occurrence, are each independently selected from:R62, R63, R67, R68for each occurrence, are each independently selected from: C1-C12 alkylene or C1-C12 alkenylene.

13. The lipid of claim 12, wherein, -R62-A3-R63-R66and -R67-A5-R68-R71independently have: a) a total number of carbon atoms selected from: 10-24, 12-22, 14-22, 16-20, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23 or 24 carbon atoms; b) a longest linear chain with a total number of carbon atoms that is selected from: 10- 24, 12-22, 14-22, 16-20, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23 or 24 carbon atoms; or c) a longest linear chain with a total number of atoms that is selected from: 10-24, 12-22, 14-22, 16-20, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23 or 24 atoms.

14. The lipid of claim 1, wherein R55, for each occurrence, is independently selected from:

15. The lipid of claim 1, having a structural formula selected from:R16for each occurrence, is independently selected from: a bond or — [CH2]k-;R62, R63, R64, R67, R68, R69, R74, R75, R76and R77for each occurrence, are each independently selected from: a bond, C1-C24 alkylene or C1-C24 alkenylene;R61and R73for each occurrence, are each independently selected from:R65, R70and R78for each occurrence, are each independently selected from: a bond, - CH=CH-CH2-CH=CH- , -CH=CH-CH2-cPr-, -cPr-CH2-CH=CH-, -cPr-CH2-cPr-, -CH=CH-CD2- CH=CH- , -CH=CH-CD2-cPr-, -cPr-CD2-CH=CH- -or -cPr-CD2-cPr-, where -cPr- is cyclopropylene, each double bond or cPr has a cis configuration and D denotes deuterium (2H); andR66, R71and R79for each occurrence, are each independently selected from: C1-C24 alkyl or Ci-C24alkenyl.

16. The lipid of claim 15, wherein: -R62-A3-R63-A4-R64-R65-R66, -R62-A3-R63-A4-R64-R73-R74-A7- R75-R65-R66, -R62-A3-R63-A4-R64-R73-R76-A8-R77-R78-R79and -R67-A5-R68-A6-R69-R70-R71, each independently has a total of 10-24 carbon atoms.

17. The lipid of claim 1, having a structural formula selected from:(12); wherein:R61for each occurrence, is independently selected from:R85and R90for each occurrence, are each independently selected from: a bond or -[CH2]I-S-R86and R91for each occurrence, are each independently selected from: a bond -O-, -O-C(O=)-, -O-C(=O)-O- or -C(=O)-O-;R87for each occurrence, is independently selected from: a bond or -[CH2]I-6-;R88and R89for each occurrence, are each independently -[CH2]I-IOCH3;R92is -[CH2]2-I6CH3;R93and R94for each occurrence, are each independently selected from: -H, -D, -CH3, -CD3, - CH2CH3, -CD2CH3, where D denotes deuterium (2H);R95, R96, R100, R101and R102for each occurrence, are each independently selected from: - OH, -O-C1-C4 alkyl, -O-C(=O)-O-Ci-C4alkyl or -O-C(=O)-CrC4alkyl;R97for each occurrence, is independently selected from: -[CH2]I-6-, -[CH2]I-6-C(=O)-O-, - [CH2] -[CH2]I-6-O- or -[CH2]I-6-C(=O)-, wherein: when R61then R97for each occurrence, is independently selected from: - [CH2] I-6-C(=O)-O- and when R61then R97for each occurrence, is independently selected from: -[CH2]-C(=O)-O-, -[CH2]I-6-O- or -[CH2]I-6-C(=O)-; andR16for each occurrence, is independently selected from: a bond or -[CH2]k-;R98for each occurrence, is independently selected from: a bond, -C(=O)-, or -C(=O)-O-; and R104for each occurrence, is independently selected from:

18. The lipid of claim 17, wherein: a) R85and R90are the same, b) R85and R90are each independently -[CH2]4-5-, c) one or both of R86and R91are / is -C(O=)-O-, d) one or both of R86and R91is / are a bond, e) R88and R89are each independently -[CH2]6-IO-, f) R92is -[CH2]8-i2-, g) R85, R86, R87, R88and R89, together, have between 14-24 or between 16-22 total carbon atoms, h) R90, R91and R92, together, have 12-18 total carbon atoms, i) R88and R89are identical,j) R86, R87and R88, together, and R90, R91and R92, together, have total number of carbon atoms that differ by + / - 3, k) R93and / or R94are / is -H, or l) R95and / or R96are / is -OH.

19. The lipid of claim 1, having a structural formula selected from:wherein:R80for each occurrence, is independently selected from: a bond, R16or -[CH2]I-6-O- or -O-;R16for each occurrence, is independently selected from: a bond or -[CH2]k-;R81, R82, R83and R84are each independently selected from: a bond or -[CH2]I-IO-;R93and R94for each occurrence, are each independently selected from: -H, -D, -CH3, -CD3, - CH2CH3, -CD2CH3, where D denotes deuterium (2H);R95and R96for each occurrence, are each independently selected from: -OH, -O-C1-C4 alkyl, -O-C(=O)-O-CrC4alkyl or -O-C(=O)-Ci-C4alkyl;R97for each occurrence, is independently selected from: -[CH2]I-6-, -[CH2]I-6-C(=O)-O-, -[CH2] -[CH2]I-6-O- or -[CH2]I-6-C(=O)-, wherein: when R61then R97for each occurrence, is independently selected from: - [CH2] I-6-C(=O)-O- and when R61then R97for each occurrence, is independently selected from: - [CH2]-C(=O)-O-, -[CH2]I-6-O- or -[CH2]I-6-C(=O)-; andR98for each occurrence, is independently selected from: a bond, -C(=O)-, or -C(=O)-O-.

20. The lipid of claim 1, wherein: a) R81and R83are each independently -[CH2]s-9, and R82and R84are each independently - [CH2]O-6, b) R81and R83are each independently -[CH2]S-IO-, and R82and R84are each independently -[CH2]O-6-, c) R81and R82, combined, have 7-13 total carbon atoms, d) R83and R84, combined, have 7-13 total carbon atoms, e) R81and R82, combined, have 8-12 total carbon atoms, f) R83and R84, combined, have 8-12 total carbon atoms, g) R81and R82, combined, and R83and R84, combined, have an identical number of total carbon atoms which is between 7-13, or h) R81and R82, combined, and R83and R84, combined, have a different number of total carbon atoms.

21. The lipid of claim 1, having a structural formula selected from:(27) or(28); wherein:R85and R90for each occurrence, are each independently selected from: a bond or-[CH2]I-8-;R86and R91for each occurrence, are each independently selected from: a bond -O-, -O- C(O=)-, -O-C(=O)-O- or -C(=O)-O-;R87for each occurrence, is independently selected from: a bond or -[CH2]I-6-;R88and R89for each occurrence, are each independently -[CH2]I-I0CH3;R92is -[CH2]2.I6CH3;R93and R94for each occurrence, are each independently selected from: -H, -D, -CH3, -CD3, -CH2CH3, -CD2CH3, where D denotes deuterium (2H);R96for each occurrence, is each independently selected from: -OH, -O-C1-C4 alkyl, -O- C(=O)-O-C1-C4alkyl or -O-C(=O)-Ci-C4alkyl;R97for each occurrence, is independently selected from: -[CH2]I_6-, -[CH2]I.6-C(=O)-O-,-[ -, -[CH2]I-6-O- or -[CH2]I-6-C(=O)-, wherein: when, then R97for each occurrence, is independently selected from:-[CH2]I-6- or -[CH2]I-6-C(=O)-O- and when R61is ~L , then R97for each occurrence, is independently selected from: -[CH2]I-6-, -[CH2]I-6-C(=O)-O-, -[CH2]I-6-O- or -[CH2]I-6-C(=O)-; andR98for each occurrence, is independently selected from: a bond, -C(=O)-, or -C(=O)-O-.

22. The lipid of claim 1, having a structural formula selected from:(29),23. The lipid of claim 1, wherein: a) R1and R3, for each occurrence, are each independently, -H, optionally substituted Cr Ce alkyl, optionally substituted C2-C6 alkenyl, optionally substituted C1-C6 hydroxyalkyl or optionally substituted C2-C6 hydroxyalkenyl, b) R1and R3, for each occurrence, are each independently selected from: -H, methyl, ethyl, propyl, isopropyl, butyl, isobutyl or tertbutyl, and R2is absent or -H or c) R1and R3for each occurrence, are each independently selected from: -H, -D, -CH3, -CD3, -CH2CH3, -CD2CH3, where D denotes deuterium (2H), and R2is absent, d) R1and R3, for each occurrence, are each independently selected from: -H, C1-C6 alkyl, C2-C6 alkenyl, C1-C5 alkyl, C2-C5 alkenyl, C1-C4 alkyl, C2-C4 alkenyl, Ce alkyl, C5 alkyl, C4 alkyl, C3 alkyl, C2 alkyl, Ci alkyl, Ce alkenyl, or C5 alkenyl, or C4 alkenyl, or C3 alkenyl, or C2alkenyl. In some further embodiments, e) R1and R3, for each occurrence, are each independently selected from: -H or C1-C3 alkyl, or f) RTand R3are identical and R2is absent.

24. The lipid of claim 1, wherein:R4for each occurrence, is independently selected from25. The lipid of claim 1, wherein: a) R6for each occurrence, is independently selected from C3-C24 branched alkylene or C3- C24 branched alkenylene; b) R6for each occurrence, is independently selected from: Ci -C3 alkylene, C1-C9 alkylene, C2-C9 alkenylene, C1-C7 alkylene, C2-C7 alkenylene, Ci -C5 alkylene, C2-C5 alkenylene, C2- C8alkylene, C2-C8 alkenylene, C3-C7 alkylene, C3-C7 alkenylene, C5-C7 alkylene, C5-C7 alkenylene, C12 alkylene, C11 alkylene, C10 alkylene, C9 alkylene, C8alkylene, C7 alkylene, C6alkylene, C5alkylene, C4alkylene, C3alkylene, C2alkylene, Ci alkylene, C12 alkenylene, C11 alkenylene, Cwalkenylene, C9 alkenylene, C8 alkenylene, C7 alkenylene, Ce alkenylene, C5 alkenylene, C4 alkenylene, C3 alkenylene, C2 alkenylene; c) R8and R9, are each independently C6-C12 alkyl or C6-C12 alkenyl; d) R8is Ce-Cio alkyl or Ce-Cio alkenyl; e) R8and R9, together, have a total of 12-20 carbon atoms; f) R8, R9, R12, and R13, each independently have 1, 2 or 3, C=C double bonds where at least one of the C=C double bonds is of Z configuration or where each C=C double bond is in a Z configuration; g) R14is C1-C3 alkyl; h) R8and R9, for each occurrence, are each independently selected from: -Cio-Cis alkenyl comprising 1, 2, 3 or 4 cis double bonds, -Cio-Cis alkenyl ester comprising 1, 2, 3 or 4 cis double bonds, optionally substituted -C6-C18 alkyl, -Ce-7i8alkyl ester, -C6-C18 alkyl ether, -C6-C18 alkyl carbonate, -C6-C18 alkenyl, -C6-C18 alkenyl ester, -C6-C18 alkenyl ether, or -C6-C18 alkenyl carbonate; i) R8and R9, are each independently -Cio-Cis alkenyl comprising 2 cis (Z configuration) double bonds. The lipid of claim 1, R8and R9for each occurrence, are each independently selected from: optionally substituted C6-C18 alkyl, Ce-7i8alkylester, Ce- Ci8 alkyl ether, C6-C18 alkyl carbonate C6-C18 alkenyl, C6-C18 alkenylester, C6-C18 alkenylether, or C6-C18 alkenylcarbonate; j) R8and R9for each occurrence, are each independently selected from: C6-C14 alkyl, C6- C14 alkenyl, C8-C12 alkyl, C8-Ci2alkenyl, Ci6alkyl, C15 alkyl, C14 alkyl, C13 alkyl, C12 alkyl, C11 alkyl, C10 alkyl, C9 alkyl, C8alkyl, C7 alkyl, Ci6 alkenyl, C15 alkenyl, C14 alkenyl, C13 alkenyl, C12 alkenyl, C11 alkenyl, C10 alkenyl, C9, alkenyl, C8alkenyl, or C7 alkenyl; k) R8and R9have an equal number of carbon atoms; l) R8and R9, combined, have more than 15 total carbon atoms; m) R8and R9differ in total carbon atoms from each other; n) R8and R9differ by one or two total carbon atoms from each other; o) R8and R9differ by one total carbon atoms; p) R8is C7 alkyl and R9is C8alkyl, R8is C8alkyl and R9is C7 alkyl, R8is C8alkyl and R9is C9 alkyl, R8is C9 alkyl and R9is C8alkyl, R8is C9 alkyl and R9is C10 alkyl ,R8is C10 alkyl and R9is C9alkyl, R8is C10 alkyl and R9is C11 alkyl, R8is C11 alkyl and R9is C10 alkyl, R8is C11 alkyl and R9is C12 alkyl, R8is C12 alkyl and R9is C11 alkyl, R8is C7alkyl and R9is C9alkyl,R8is C9 alkyl and R9is C7 alkyl, R8is C8alkyl and R9is C10 alkyl, R8is C10 alkyl and R9is C8alkyl, R8is C9 alkyl and R9is C11 alkyl, R8is C11 alkyl and R9is C9 alkyl, R8is C10 alkyl and R9is C12 alkyl, R8is C12 alkyl and R9is C10 alkyl, R8is C11 alkyl and R9is C8alkyl, or R8is C8alkyl and R9is C11 alkyl; q) R10and R11for each occurrence, are each independently selected from: C1-C16 unbranched alkyl or C2-C16 unbranched alkenyl; r) R16is C1-C3 alkylene; s) R29, R30and R31are each -CH2-; t) R32, R33and R34are each -O-; u) R35is -OH; v) R29, R30and R31are each -CH2-, and R32, R33and R34are each -O-; w) R29, R30and R31are each -CH2-, R32and R33are each -O- and R35is -OH; x) R42is -[CH2]nC(=O)O[CH2]r; wherein r is an integer selected from 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16 or 17; y) R42for each occurrence, is independently selected from: -R46-C(=O)O-R47-, -R46- OC(=O)-R47-, -R46-OC(=O)O-R47-,or -R46-O-R47-, wherein; R46for each occurrence, is independently selected from C1-C12 alkylene, C1-C12 alkylene ester, C1-C12 alkylene ether, C1-C12 alkylene carbonate; and R47for each occurrence, is independently selected from -C1-C12 alkyl, -C1-C12 alkyl ester, -C1-C12 alkyl ether, or -C1-C12 alkyl carbonate.

26. The lipid of claim 1, wherein: R55for each occurrence, is independently selected from:R5for each occurrence, is independently selected from: -H, optionally substituted C1-C16 alkyl, optionally substituted C1-C16 alkyl ester, optionally substituted C2-C16 alkenyl, optionally substituted C2-C16 alkenyl ester,; wherein, R10and R11for each occurrence, are each independently selected from: optionally substituted C1-C16 alkyl or optionally substituted C2-C16 alkenyl.T1. The lipid of claim 26, wherein: R5for each occurrence, is independently selected from: C1-C14 unbranched alkyl, C2-C14 unbranched alkenyl, C1-C16 unbranched alkyl, C1-C16 unbranched alkyl ester, unbranched C2-Ci6alkenyl, C2-Ci6unbranched alkenyl ester, C3-C16 branched alkyl, C3-C16 branched alkyl ester, C3-C16 branched alkenyl, C3-C16 branched alkenyl ester or, wherein: R10and R11for each occurrence, are each independently selected from: C1-C12 unbranched alkyl, C2-C12, unbranched alkenyl, C2-Ci2unbranched alkyl, C2- Ci2unbranched alkenyl, C5-C7, unbranched alkyl, or C5-C7 unbranched alkenyl, Ci6 unbranched alkyl, C15 unbranched alkyl, C14 unbranched alkyl, C13 unbranched alkyl, Ci2unbranched alkyl, C11 unbranched alkyl, C10 unbranched alkyl, C9unbranched alkyl, C8unbranched alkyl, C7 unbranched alkyl, Ce unbranched alkyl, C5 unbranched alkyl, C4 unbranched alkyl, C3 unbranched alkyl, C2unbranched alkyl, methyl, Ci6 unbranched alkenyl, C15 unbranched alkenyl, C14 unbranched alkenyl, C13 unbranched alkenyl, C12 unbranched alkenyl, C11 unbranched alkenyl, C10 unbranched alkenyl, C9unbranched alkenyl, C8unbranched alkenyl, C7unbranched alkenyl, C6unbranched alkenyl, C5unbranched alkenyl, C4unbranched alkenyl, C3unbranched alkenyl, C2alkenyl, C2-Ci0unbranched alkyl or C2-C10 unbranched alkenyl.

28. The lipid of claim 1, wherein: a) R55for each occurrence, is independently selected from: -OC(=O)C(R42)[CH2]nC(=O)O- R41-CH(C4-CI2alkyl)2, -R41-N(R42)([CH2]nC(=O)O-R41-CH(C4-Ci2alkyl)2, - OC(=O)C(R43)[CH2]nC(=O)O-R41-CH(C4-Ci2alkyl)2, -R41-N(R43)([CH2]nC(=O)O-R41-CH(C4- Ci2alkyl)2, -R41-C(R42)[CH2]nC(=O)O-R41-CH(C4-Ci2alkyl)2, -R41-CH(R43)(R44), or -R41- N(R43)(R44); b) A2for each occurrence, is independently selected from; -OC(=O)-, -OC(=O)R16C(=O)O- or -C(=O)O-; c) each R55has 12-50, 18-30, 20-30, or 12-24 total carbon atoms; and d) each R55comprises 0-6, 0-3, 1-3, 1, 2, 3, 4, 5 or 6 esters; 0-6, 0-3, 1-3, 1, 2, 3, 4, 5 or 6 ethers; and / or 0-6, 0-3, 1-3, 1, 2, 3, 4, 5 or 6 carbonates.

29. The lipid of claim 1, R55for each occurrence, is independently selected from: -O- C(=O)C(R42)[CH2]nC(=O)O-R41-CH(C4-Ci2alkyl)2, -R41-N(-R42)(-[CH2]nC(=O)O-R41-CH(C4- Ci2alkyl)2, -O-C(=O)C(R43)-[CH2]nC(=O)O-R41-CH(C4-Ci2alkyl)2, -R41-N(-R43)(- [CH2]nC(=O)O-R41— CH(C4-CI2alkyl)2, -R41-C(R42)[CH2]nC(=O)O-R41-CH(C4-Ci2alkyl)2, -R41- CH(R43)(R44), or -R41-N(R43)(R44).

30. The lipid of claim 1, wherein: R55, for each occurrence, is independently selected from: -R16-N(-R48)-R49, -R16-CH(-R48)-R49, -R16-OC(=O)-R16-N(-R48)-R49, -R16-OC(=O)CH(-R48)-R49, or -R16-C(=O)O-R16-CH(-R48)-R49; wherein:R16for each occurrence, is independently selected from: a bond or -[CH2]k-; andR48and R49are selected from : i) R48is -R16-O-CH(-[CH2]5-9 CH3)2and R49is -R16-C(=O)O-[CH2]6-I2CH3, j) R48is -CH2O-C(=O)O-CH(-[CH2]7CH3)2and R49is -R16-O-C(=O)O-[CH2]7-I3CH3, k) R48is -R16C(=O)O-CH([CH2]7CH3)2and R49is -R16C(=O)O-[CH2]7CH3, l) R48is -[CH2]3C(=O)O-CH(-[CH2]7CH3)2and R49is -[CH2]3C(=O)O-[CH2]7CH3, m) R48is -CH2C(=O)O-CH(-[CH2]9CH3)2and R49is -C(=O)O-[CH2]I0CH3, n) R48is -[CH2]3C(=O)O-CH(-[CH2]7CH3)2and R49is -[CH2]3C(=O)O-[CH2]9CH3 o) R48is -[CH2]2C(=O)O-CH(-[CH2]7CH3)2and R49is -[CH2]2C(=O)O-[CH2]9CH3, or p) R48is -R16-C(=O)O-R16-CH(-R57-CH3)2and R49is -[CH2]8-I6CH3.

31. The lipid of claim 1, wherein: R55, for each occurrence, is independently selected from:each cyclopropyl (cPr) and each -C=C- has a cis (or E) configuration;R57for each occurrence, is independently selected from: a bond or -[CFhJm-; wherein m is an integer selected from: 1, 2, 3, 4, 5, 6, 7 or 8;R16for each occurrence, is independently selected from: a bond or -[CFhJk-;R61for each occurrence, is independently selected from:R116for each occurrence, is independently selected from: -H, -F, -CF3, -Cl, -Br, -I, -CH3, - CH2R14, -CHR142, -CR143, -OH, -OR14, -NH2, -NHR14, -N(R14)2, -SH, -SR14, -S(=O)H, - S(=O)R14, -S(=O)2H, -S(=O)2R14, -PH2, -PHR14, -PR142, -P(OH)2, -POHR14, -PR142, -P(OR14)2, -P(O)H2, -P(O)HR14, -P(O)R142, -P(O)(OH)2, -P(O)(OR14)OH, -P(O)(OR14)2, -SiH3, -SiH2R14, - SiHR142or-SiR143;R81, R82, R83and R84for each occurrence, are each independently selected from: a bond or -[CH2]I-IO-;R85and R90for each occurrence, are each independently selected from: a bond or -[CH2]I-S-R86and R91for each occurrence, are each independently selected from: a bond -O-, -O- C(O=)-, -O-C(=O)-O- or -C(=O)-O-;R87for each occurrence, is independently selected from: a bond or -[CH2]I-6-;R88and R89for each occurrence, are each independently -[CI-hJi-ioCHs;R92is -[CH2]2-I6CH3;R97for each occurrence, is independently selected from: -[CH2]I-6-, -[CH2]I-6-C(=O)-O-, - [CH2] -[CH2]I-6-C(=O)-, wherein: when R61then R97for each occurrence, is independently selected from: -[CH2]I-6- or -[CH2]I-6-C(=O)-O- and when R61is ~L , then R97for each occurrence, is independently selected from: - [CH2]I-6-, -[CH2]I-6-C(=O)-O-, -[CH2]I-6-O- or -[CH2]I-6-C(=O)-.

32. A lipid particle comprising the lipid if claim 1.

33. The lipid particle of claim 32, wherein said particle is a lipid nanoparticle or liposome.

34. The lipid nanoparticle of claim 33, wherein the lipid component further comprises one or more components selected from the group consisting of: a neutral lipid, a structural lipid and a polymer conjugated lipid.

35. The lipid nanoparticle of claim 34, wherein the neutral lipid is selected from one or more of the group consisting of: l,2-distearoyl-sn-glycero-3-phosphocholine (DSPC), 1,2- dioleyl- sn-glycero-3-phosphoethanolamine (DOPE), l,2-dilinoleyl-sn-glycero-3- phosphocholine (DLPC), 1,2-dimyristoyl-sn-glycero-phosphocholine (DMPC), 1,2- dioleyl- sn-glycero-3-phosphocholine (DOPC), l,2-dipalmitoyl-sn-glycero-3-phosphocholine (DPPC), 1,2-diundecanoyl-sn-glycero-phosphocholine (DUPC), 1- palmitoyl-2-oleyl-sn- glycero-3-phosphocholine (POPC), 1,2-di-O-octadecenyl-sn- glycero-3-phosphocholine (18:0 Diether PC), l-oleyl-2-cholesterylhemisuccinoyl-sn- glycero-3-phosphocholine (OChemsPC), l-hexadecyl-sn-glycero-3-phosphocholine (C16 Lyso PC), 1,2-dilinolenoyl- sn-glycero-3-phosphocholine, 1,2-diarachidonoyl-sn- glycero-3-phosphocholine, 1,2- didocosahexaenoyl-sn-glycero-3-phosphocholine, l,2-diphytanoyl-sn-glycero-3- phosphoethanolamine (ME 16.0 PE), l,2-distearoyl-sn-glycero-3-phosphoethanolamine, l,2-dilinoleyl-sn-glycero-3-phosphoethanolamine, l,2-dilinolenoyl-sn-glycero-3-phosphoethanolamine, l,2-diarachidonoyl-sn-glycero-3-phosphoethanolamine, 1 ,2- didocosahexaenoyl-sn-glycero-3-phosphoethanolamine, 1 ,2- dioleyl-sn-glycero-3- phospho-rac-(l-glycerol) sodium salt (DOPG), and sphingomyelin.

36. The lipid nanoparticle of claim 34, wherein the structural lipid is selected from one or more of the group consisting of: cholesterol, fecosterol, sitosterol, campesterol, stigmasterol, brassicasterol, ergosterol, tomatidine, tomatine, ursolic acid and alphatocopherol, 5-a-cholestanol (5a-Cholestan-3|3-ol), 5-0-coprostanol, cholesteryl-(2' - hydroxy)-ethyl ether, cholesteryl-( 4' -hydroxy)-butyl ether, and 6-ketocholestanol, 5a- cholestane, cholestenone, 5-a-cholestanone, 5 p-cholestanone, allocholesterol, epiallocholesterol, cholesteryl decanoate, cholesteryl-(4'-hydroxy)-butyl ether, cholesteryl hemisuccinate, cholest-5-en-3|3-yl hydrogen sulfate and 24-methylene-cholesterol sulfate.

37. The lipid nanoparticle of claim 34, wherein the polymer conjugated lipid is selected from one or more of the group consisting of: PEGylated lipid selected from the group consisting of PEG-modified phosphatidylethanolamines, PEG-modified phosphatidic acids, PEG-modified ceramides, PEG-modified dialkylamines, PEG-modified diacylglycerols, and PEG-modified dialkylglycerols, optionally PEG-c-DOMG, PEG-DMG, PEG-DLPE, PEG-DMPE, PEG-DPPC, and PEG-DSPE.

38. The lipid nanoparticle of claim 34, comprising: about 25 mol % to about 60 mol % of a lipid selected from any one or more of Compound (1) - (54); about 2 mol % to about 25 mol % neutral lipid; about 18.5 mol % to about 60 mol % structural lipid; and about 0.2 mol % to about 10 mol % of PEGylated lipid.

39. The lipid nanoparticle of claim 34, wherein the lipid nanoparticle further comprises a diagnostic, prophylactic or therapeutic payload selected from a therapeutic nucleic acid, a protein a peptide or a small molecule.

40. The lipid nanoparticle of claim 39, wherein the therapeutic nucleic acid is an mRNA or a self-amplifying RNA.

41. The lipid nanoparticle of claim 34, wherein the lipid nanoparticle has a diameter of from about 30 nm to about 160 nm.

42. A pharmaceutical lipid nanoparticle composition comprising the lipid nanoparticle of claim 34 and a pharmaceutically acceptable carrier, excipient or diluent.

43. A method of delivering a lipid nanoparticle of claim 34 to a mammalian cell, comprising contacting the cell with the lipid nanoparticle.

44. The method of item 43, wherein the cell is a cell of a human subject.

45. A method of treating a disease, disorder or condition in a subject in need of such treatment, comprising administering a therapeutically effective amount of the pharmaceutical composition of claim 42 to the subject to thereby treat the disease, disorder or condition.

46. Use of a lipid or lipid nanoparticle of any one of claims 32-41 or the lipid nanoparticle composition of claim 11 in the manufacture of a medicament for the treatment of a disease, disorder or condition.

47. The method of claim 45, wherein the disease, disorder or condition is selected from the group consisting of a rare disease, an infectious disease, cancer, a proliferative disease, a genetic disease, an autoimmune disease, diabetes, a neurodegenerative disease, a cardiovascular disease, a reno-vascular disease and a metabolic disease.

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