Lipids and lipid nanoparticles

Abstract

The disclosure provides for lipids that may be formulated in a delivery vehicle to facilitate the encapsulation of a wide range of single or multiple payloads including therapeutic, theragnostic, preventive, prophylactic, pre-emptive, and diagnostic agents, such as, without limitation, nucleic acids (e.g., RNA or DNA), proteins, peptides, and small molecule active pharmaceutical ingredients (APIs). Methods of delivering and / or producing a polypeptide of interest in a cell are also provided.

Description

Lipids and Lipid NanoparticlesBackground 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 but not limited to nucleic acids such as DNA, RNA, circRNA, self-amplifying RNA, small-activating 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. While various molecules of different sizes and properties can be encapsulated in lipid particles, the 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 delivery and 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 and safety in combating COVID-19, proving themselves as life-saving technology. These vaccines not only showcased the potential of LNPs but also heralded the development and deployment of mRNA-based vaccines and therapeutic 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 the reticuloendothelial 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 unwanted immune responses. LNPs may induce an immune response due to their foreign nature and inherent adjuvant potential, 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.Considering the structural importance of lipids and their influence in the overall particle biophysical characteristics, there exists a need to develop new lipid compounds, e.g., cationic lipids, ionizablelipids (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), preferential if not selective targeting to cells / tissues / organs, 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 single or multiple payloads including therapeutic, theragnostic, preventive, prophylactic, pre-emptive, 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 DrawingsFIG. 1. The Iog2 fold change of whole-body bioluminescence of firefly luciferase of LNPs containing novel ionizable lipids over LP-O1 LNPs in mouse. The in vivo experiments were done in C57BL / 6 mouse at 0.25 mg / kg and the bioluminescence was collected at t=6 hr.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 theragnostic, preventive, prophylactic, pre-emptive therapeutic, pharmacokinetic, diagnostic, and theragnostic purposes, or companion medicine and 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, but not limited to, oral, pulmonary, intranasal, parenteral (e.g., intravenous, intramuscular, intraperitoneal, or subcutaneous), rectally, intravesical, intranodal, intralymphatical, intratumoral, regional, local and topical administration."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 negativeIog(10) value of the apparent acid dissociation constant (Ka) of the protonated, positively charged form of the lipid in the LNP assembly context. Cationic and cationic ionizable lipids are highly waterinsoluble, even when charged, due to the lipophilic nature of the overall molecule. 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 plus charged 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 "noncationic lipid" is an anionic or neutral lipid."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."Hydrocarbylene" refers to a group formed by removing two hydrogen atoms from a hydrocarbon, the free valencies of which are not engaged in a double bond."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."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 notlimited 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 Ci-Cg 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)."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-l V), 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, pre-condensed DNA, PCR products, vectors (PI, PAC, BAC, YAC, artificial chromosomes), expression cassettes, chimeric sequences, chromosomal DNA, or derivatives and combinations 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), circRNA, self-amplifying RNA, small-activating RNA, long nocoding RNA, short non-coding RNA, 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-naturallyoccurring, 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 theTNA 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, FISH, RNAscope™, 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."Amine" or "amino" as used herein interchangeably refers to a functional group that contains a basic nitrogen atom with a lone pair."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 more cyclic rings in which two or more carbons are common to two adjoining rings, wherein at least one of the rings is aromatic. A "carboaryl" refer to an aryl group containing only carbon and hydrogen atoms."Arylene" 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."Aralkyl" refers any alkyl group (as defined herein) in which one or more hydrogen atoms have been substituted by the same number of like and / or different aryl (as defined herein) groups.The term "arenyl" means any aryl group (as defined herein) in which one or more hydrogen atoms have been substituted by the same number of like and / or different alkyl groups (as defined herein).The term "alkene" refers to a group consisting of at least two carbon atoms and at least one carboncarbon 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."Alkenyl" refers to a hydrocarbon monovalent 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 carbon-carbon double bonds (i.e., a group derived from an alkene with two attachment points), 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. "Internal alkenylene" refers to an alkenylene group where the carbon-carbon double bond is not at an end of the carbon chain, i.e., the terminal carbon atoms are not double bonded."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 carbon-carbon triple bonds (i.e., a group derived from an alkyne with two attachment points). "Internal alkynylene" refers to an alkynylene group where the carbon-carbon triple bond is not at an end of the carbon chain, i.e., the terminal carbon atoms are not triple bonded.Cyclic groups of the present invention include monocyclic and bicyclic groups (bridged or fused)."Alkenyl ester" refers to - alkylene -C(=O)O-R53or - alkylene -OC(=O)-R53, wherein R52is alkylene or alkenylene and R53is alkyl or alkenyl, with the proviso that R52and / or R53contain at least one carboncarbon double bond."Alkenyl carbonate" to -R52-O-C(=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."Alkyl" refers to saturated monovalent hydrocarbon radical. Alkyls may be linear or branched and may be optionally substituted. Examples include Ci.2Oalkyl, Ci-i8alkyl, Ci-i6alkyl, C1.14 alkyl, Ci-i2alkyl, CMO alkyl, Ci-9 alkyl, Ci.g alkyl, C1-7 alkyl, Ci.g 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."Alkylene" refers to a saturated bivalent hydrocarbon radical (a group derived from an alkane with two attachment points). Alkylenes may be linear or branched and may be optionally substituted. Examples include Ci-2o alkylene groups, such as Ci-2o alkylene, Ci-ig alkylene, CUB alkylene, C1-14 alkylene, Ci-i2alkylene, C1-10 alkylene, C1-9 alkylene, Ci-g alkylene, C1-7 alkylene, CI-B alkylene, C1-5 alkylene, C1-4 alkylene, and C1.C3 alkylene.The term "alkyl ester" refers to -alkylene-C(=O)O-alkyl or -alkylene-OC(=O)-alkyl.The term "alkyl carbonate" refers to - alkylene-O-C(=O)O-alkyl."Bivalent" as used herein refers a functional group with two attachment points. Bivalent groups are formed by the loss of a hydrogen atom from two different atoms of a parent compound and are named using the suffixes -diyl or -ylene."Carbocycle" and "carbocyclic" refers to a C3-C2o 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."-cBu-" is cyclobutyl, "-cBu-" is cyclobutylene; ; "-ePt" is cyclopentyl, "-ePt-" is cyclopentylene, "-cHx" is cyclohexyl, "-cHx-" is cyclohexylene, "-cPr" is cyclopropyl (cyclopropanyl), "-cPr- " is cyclopropylene (cyclopropan-1,2 diyl)."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 groupsinclude, but are not limited to: cyclopropyl, cyclobutyl, cyclopentyl, cyclopentenyl, cyclohexyl, cyclohexenyl, cycloheptyl, cyclooctyl, cyclononyl, cyclodecyl or 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."D" is deuterium (2H)."Halogen" or "halo" 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 spiro[cycloalkyl] 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 ringscan 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. 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. Stereochemistry of double bounds may be E or Z (cis or trans).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.LipidIn 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 an 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, and modified version of the above with lipid and / or carbohydrate groups or other functionalization.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. TNA can be a single kind or multiple types in the same particle system.LNP ComponentsAccording to some embodiments, the lipid particles of the disclosure have a mean diameter of: from about 40 nm to about 45 nm, from about 45 nm to about 50 nm, from about 50 nm to about 55 nm, from about 55 nm to about 60 nm, from about 60 nm to about 65 nm, from about 65 nm to about 70 nm, from about 70 nm to about 75 nm, from about 75 nm to about 80 nm, from about 80 nm to about 85 nm, from about 85 nm to about 90 nm, from about 90 nm to about 95 nm, from about 95 nm to about 100 nm, from about 100 nm to about 105 nm, from about 105 nm to about 110 nm, from about 110 nm to about 115 nm, from about 115 nm to about 120 nm, from about 25 nm to about 125 nm, from 25 nm to about 100 nm, from about 40 nm to about 200 nm, from about 40 nm to about 150 nm, from about 40 nm to about 125 nm, from about 40 nm to about 100 nm, from about 40 nm to about 90 nm, from about 40 nm to about 80 nm, from about 40 nm to about 70 nm, from about 40 nm to about 60 nm, from about 50 nm to about 80 nm, from about 50 nm to about 90 nm, from about 50 nm to about 100 nm, from about 50 nm to about 110 nm, from about 50 nm to about 120 nm, from about 50 nm to about 125 nm, from about 50 nm to about 150 nm, from about 50 nm to about 180 nm, from about 100 nm to about 150 nm, less than about 150 nm, less than about 125 nm, less than about 100 nm, less than about 80 nm less than about 75 nm, less than about 70 nm, less than about 60 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 quasielastic 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.Cationic lipidIn one aspect, the LNP comprises: a (ionizable) cationic lipid, a sterol or a derivative thereof, a noncationic 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 non-cationic 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 or 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 49-50mol%, 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% non-cationic 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 lipid is neutral at physiological pH (pH 7.4). The ability of these lipids to ionize at low pH is believed to aid 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 some embodiments, 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 lipids that may be used with a composition of the present invention are described in International PCT patent publications WO 2025 / 119217, WO 2025 / 113654, WO 2025 / 113662, WO2025 / 114520, WO 2025 / 113656, WO 2025 / 117732, WO 2025 / 111454, WO 2025 / 104695, WO 2025 / 100828, WO 2025 / 089790, WO 2025 / 082973, WO 2025 / 076625, WO 2025 / 081002, WO 2025 / 080867, WO 2025 / 080555, WO 2025 / 076127, WO 2025 / 100737, WO 2025 / 061967, WO 2025 / 066954, WO 2025 / 059599, WO 2025 / 029927, WO 2025 / 016446, WO 2025 / 012376, WO 2025 / 014802, WO 2025 / 008510, WO 2025 / 010290, WO 2025 / 003759, WO 2024 / 259531, WO 2025 / 001900, WO 2024 / 259373, WO 2024 / 256457, WO 2024 / 256453, WO 2024 / 251108, WO 2024 / 249954, WO2024 / 243480, WO2024 / 243031, WO2024 / 240133, WO2024 / 240125,WO2024 / 236361, WO2024 / 234063, W02024 / 234006, WO2024 / 233750, WO2024 / 233387, WO2024 / 232832, WO2024 / 230844, WO2024 / 226958, WO2024 / 226779, WO2024 / 223911, WO2024 / 222243, WO2024 / 220625, W02024 / 220609, WO2024 / 216171, WO2024 / 212958,WO2024 / 205657, WO2024 / 198497, WO2024 / 195922, WO2024 / 192528, WO2024 / 192277, W02024 / 184500, WO2024 / 183821, WO2024 / 177426, WO2024 / 177424, WO2024 / 177282, WO2024 / 173307, WO2024 / 165974, WO2024 / 165973, WO2024 / 156291, WO2024 / 152512, WO2024 / 150222, W02024 / 147060, W02024 / 144009, WO2024 / 138134, WO2024 / 138034,WO2024 / 136309, WO2024 / 136254, WO2024 / 135604, WO2024 / 130421, W02024 / 130086,WO2024 / 129982, WO2024 / 128525, WO2024 / 123978, WO2024 / 119098, WO2024 / 119037,WO2024 / 109929, WO2024 / 109612, WO2024 / 102762, WO2024 / 044728, WO2023 / 196931,WO2023 / 122752, WO2023 / 081526, W02023 / 056033, WO2023 / 044343, WO2022 / 261490,WO2022 / 251665, WO2022 / 246571, WO2021 / / 236855, WO2021 / 226597, WO2021 / 204179,WO2021 / 189059, WO2021 / 113777, W02021 / 077067 , WO2020 / 237227 , WO2019 / 236673,WO2019 / 232095, WO2019 / 152557, WO2019 / 089828, 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,W02005 / 120152; 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and US patents, US12102720, US12059477, US11969506, US11964052, US11851389, US11786609, US11547666, US11453639, US11406706, US11246933, US11242311, US10888626, US10556018, US10342761, US10286083, US10137086 US9896413, US9868692, US9546128, US9504747 and US9308267; each of which (each PCT publication, each US patent application and each US patent) are incorporated by reference in their entireties.Further examples include 3-(didodecylamino)-Nl,Nl,4-tridodecyl-l-piperazineethanamine (KL10), 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 or cationic lipid is represented by a structural formula selected from:wherein,R1 is selected from the group consisting of:R19 for each occurrence is independentlyR20 is R19 or R24;R21 is selected from: a bond, -CH2-, -[CH2]2-, -C(=O)-, -S-, -S(=O)- or -S(=O)2-;R22 is selected from: a bond (absent), optionally substituted Ci-Cg alkylene, Ci-Cio alkyleneoxy, -C(=O)- , -R143-CO-C3alkylene-R146-C0-C3alkylene-, -CH(OH)-, -C(OH)2-, -C(R25)2-, -CH(OR25)-, -C(OR25)2-, - N(R25)-, -N(R116)C(=O)-, -C(=O)N(R116)-, -O-, -S-, -S(=O)-, -S(=O)2-, -P(OR25)- or -P(O)(OR25)-;R116 is -H, -D, -CD3or C1-3 alkyl;R143 is selected from: a bond, -O-, -CH(OH)-, -C(OH)2-, -C(=O)-, -C(=O)O-, -OC(=O)-, -OC(=O)O-, - N(R116)-, -N(R116)C(=O)-, -C(=O)N(R116)-, -S-, -S-S-, -C(=S)-, -C(=O)-C-,-CH(OH)-, -S-C(=O)-, or -C(=O)- S-;R146 is an optionally substituted group selected from: 3-7 membered carbocyclylene or 3-7 membered heterocyclylene;R25, for each occurrence, is independently selected from: a bond, -H, -D, -CH3, -OCH3, -CD3, -OCD3, - CECH, -CH2CH3, -F, -Cl, -Br, -I, -OCH3, -C(R114)3, -NHC(=O)R114, -OR114, -N(R114)2, -SR114, -S(=O)R114, -S(=O)2R114, -P(R114)2, -P(OR114)2, -P(O)(R114)2, -Si(R114)3, -Si(O)2R114, or an optionally substituted group selected from: linear or branched C1-C10- alkyl, linear or branched C2-CM alkenyl, linear or branched C2-Cio alkynyl, a 3-18-membered saturated or partially unsaturated carbocyclyl or heterocyclyl, or a 5-18-membered unsaturated carbocyclyl or heterocyclyl; wherein said heterocyclyl contains one or more heteroatoms independently selected from N, O or S as a ring atom;R114, for each occurrence, is independently selected from: -H, -CH2(halo), -CH(halo)2, -C(halo)3, -F, - CF3, -Cl, -Br, -I, -OH, or an optionally substituted, linear or branched, group selected from C1-C6 alkyl, C2-C6 alkenyl or C2-C6 alkynyl;R23 is an optionally substituted group selected from: a 3-18-membered saturated or partially unsaturated carbocyclyl, a 5-18-membered unsaturated carbocyclyl, a 3-18-membered saturated orpartially unsaturated heterocyclyl, or 5-18-membered unsaturated heterocyclyl; wherein said heterocyclyl contains one or more of heteroatoms independently selected from N, O or S as a ring atom;R24 for each occurrence, is independently selected from: a bond, -H, -D, -CH2(halo), -CH(halo)2, - C(halo)3, -F, -CF3, -Cl, -Br, -I, -C(R25)3, -OR25, -N(R25)2, -SR25, -S(=O)R25, -S(=O)2R25, -OS(=O)2NHR25, -S(=O)2NHR25, -P(R25)2, -P(R25)2, -P(OR25)2, -P(O)(R25)2, -P(O)(OR25)2, -Si(R25)3or an optionally substituted group selected from: linear or branched C1-C10- alkyl, linear or branched C2-Ci0alkenyl, linear or branched C2-CM alkynyl;R2 is a bond, or a linear, branched or optionally substituted group selected from: -C1-C15 acyclic hydrocarbylene-, Ci-C8alkylene, C2-C8alkenylene, C2-C8alkynylene, -C0-C8acyclic hydrocarbylene- R148-CO-C8acyclic hydrocarbylene-, 3-7 membered carbocyclylene or 3-7 membered heterocyclylene;R148 is selected from: R47, R143, R146, -CH(R147)-, -O-, -S-, -S-S- or -N(R114)-;R47, for each occurrence, is independently selected from: a bond, an optionally substituted 3-14- membered saturated, partially unsaturated or unsaturated carbocyclene or an optionally substituted 3-14-membered saturated, partially unsaturated or unsaturated heterocyclene;R147, for each occurrence, is an optionally substituted group selected from: 3-7 membered carbocyclyl or 3-7 membered heterocyclyl; each R50 is independently selected from:-R62-R123-R63-R66,-R62-R123-R63-R74-R66,-R62-R123-R63-R124-R64-R65-R66,-R62-R123-R63-R124-R64-R74-R66, -R62-R123-R63-R124-R64-R74-R127-R66,-R62-R123-R63-R124-R64-R74-R127-R75-R66, or-R62-R123-R63-R124-R64-R74-R127-R75-R65-R66; wherein,R3, R4, Rll and R12 are each independently selected from: a bond, R26-Ci-Cio alkylene-R27, R26-C2- Cio alkenylene-R27, R26-C2-CIO alkynylene-R27, R26-Cg-Cio carbocyclylene-R27, R26-Cg-Cio heterocyclylene-R27R26-CH=CH-CH2-R47-R27, R26-R47-CH2-CH=CH-R27, R26-R47-CH2-R47-R27, R26- CH=CH-CD2-R47-R27, R26-R47-CD2-CH=CH-R27 or R26-R47-CD2-R47-R27; wherein, R47 for each occurrence, is an optionally substituted group independently selected from 3-10-membered carbocyclylene or 3-10-membered heterocyclylene;R26 and R27 are each independently selected from: a bond, or an optionally substituted, linear or branched, C1-C10 alkylene, C1-C10 alkenylene or C1-C10 alkynylene;R5, R6, R9 and RIO are each independently selected from: a bond, -CH2-, -O-, -S-, -CH(OH)-, -CH(NH2)- , -NH-C(=O)-O-, -O-C(=O)-NH-, -C(NH[R25])-, -C(NH[OR25])-, -C(=O)-, -O-C(R25)2-, -O-C(=O)-, -C(=O)-O- , -C(=S)-, -N(R25)-, -S-S-, -S-CH(CH3)2-S-; -C(F)2-C(=O)-O- or -O-C(=O)-C(F)2-;R7 and R8 are each independently selected from: a bond, R3, -CH2-, -C(=O)-, -C(=S)- -CH(R25)-, - C(R25)2-, -CH(OR25)- , -CH(NH[R25])-, -C(NH[OR25])-, -N(R25)-, -O-, -S-, -S(OH)-, -S(=O)-, -S(=O)2-, - S(OH)2-, -C(=O)C(R25)2C(=O)-, -C(R25)2C(=O)-, -C(=O)C(R25)2-O-, -C(R25)2C(R25)2-, -C(R25)2-O-, -O- C(R25)2C(R25)2-, -O-C(R25)2C(=O)-, -C(=O)-O-R115-O-C(=O)-, -P(OH)-, -P(OH)(R25)-, -P(=O)(OH)-, - P(OR25)2-, -P(OR25)-, -P(=O)(OR25)-, -C(F)2C(=O)-O- or -O-C(=O)C(F)2-;R115, for each occurrence, are each independently selected from: R26-Ci-Cg alkylene-R27, R26-C2-C6 alkenylene-R27, R26-C2-C8alkynylene-R27, R26-R47-R27, R26-CH=CH-CH2-R47-R27, R26-R47-CH2- CH=CH-R27, R26-R47-CH2-R47-R27, R26-CH=CH-CD2-R47-R27, R26-R47-CD2-CH=CH-R27 or R26-R47- CD2-R47-R27; wherein, R47 for each occurrence, is an optionally substituted group independently selected from 3-10-membered carbocyclylene or 3-10-membered heterocyclylene; and each double bond or cPr has a cis configuration;R13, R14, R61, R73 and R107 are each independently selected from:R60 is selected from: is absent (i.e., an unbonded pair of electrons on N), -H, -D -CD3or optionally substituted C1-C6 alkyl; wherein, when R60 is -H or optionally substituted C1-C6 alkyl, R60 is bound directly to a nitrogen atom with a +1 charge. The +1 charge may be counterbalanced with anion from a pharmaceutically acceptable acid;R15, R16, R17 and R18 are each independently selected from: -H, -D; an optionally substituted linear or branched, -R26-Ci-Cio alkylene-R28, -R26-C2-C10 alkenylene-R28, or -R26-C2-C10 alkynylene-R28; an optionally substituted, R26-R149, R26-R23, -R26-C3-C7cycloalkylene-R28, -R26-C3-C7cycloalkylene- R26-C3-C7cycloalkylene-R28 -R26-C3-C7cycloalkenylene-R28, -R26-C5-Ci5-spirocycloalkylene-R28, - R26-C3-Ci5-carbocyclylene-R28, -R26-C3-Ci5-heterocyclylene-R28, -R26-arylene-R28, or -R26- heteroarylene-R28; or -R26-CH=CH-CH2-cPr-R28, -R26-cPr-CH2-CH=CH-R28, -R26-cPr-CH2-cPr-R28, - R26-CH=CH-CD2-R47-R28, -R26-R47-R47-R28, -R26-R47-R28, -R26-R47-R26-R47-R28, -R26-CD2-R47- R28, -R26-R47-CD2-R47-R28, -R26-R47-CD2-CH=CH-R28, -R26-CH=CH-CH2-CH=CH-R28, -R26-CH=CH- CH2-R47-R28, -R26-R47-CH2-CH=CH-R28, -R26-R47-CH2-R47-R28, -R26-CH=CH-CD2-CH=CH-R28;R28 is selected from: -H, -D, -CDs, C1-C10 alkyl, C2-C10 alkenyl or C2-C10 alkynyl;R149 is:; wherein, each R155 is each independently selected from: -H, -OH, optionally substituted C1-C6 alkyl, optionally substituted C2-C6 alkenyl, optionally substituted C2-C6 alkynyl or optionally substituted C1-C6 alkoxy;R150 is selected from optionally substituted -CD3, CI-C15 alkyl, optionally substituted C2-Ci5alkenyl, optionally substituted C2-C15 alkynyl, optionally substituted C1-C15 alkoxy, optionally substituted C1-C10 alkylene-C(O)O-optionally substituted C1-C15 alkyl or optionally substituted Ci-Cioalkylene-COOH;R151 is -H or -D and R152 is -OH, or R151 and R152 are both -H, or R151 and R152 are both absent; when R151 and R152 are absent a C=C double bond is present between the carbon atoms that R151 and R152 are bound to; and,R153 is -OH and R154 is -H or -D, or R153 and R154 are both -H or -D, or R153 and R154 are both absent, wherein when R153 and R154 are absent then a C=C double bond, preferably cis (Z configuration stereochemistry) is present between the carbon atoms to which R153 and R154 are bound;R62, R63, R64, R67, R68, R69, R74, R75, R76, R77, R108, R109, RllO and Rill are each independently selected from: a bond, C1-C10 alkylene, C2-C10 alkenylene, C2-C10 alkynylene, C3-C7 cycloalkylene, C3-C7 cycloalkenylene, C5-Ci0-spirocycloalkylene, C3-Ci0-carbocyclylene, C3-Ci0-heterocyclylene, -R26- CH=CH-C2-C6alkylene-CH=CH-R27-, -R26-CH=CH-C2-C6alkylene-R47-R27-, -R26-R47-C2-C6alkylene- CH=CH-R27-, -R26-R47-C2-C6alkylene-R47-R27-, -R26-C3-C6-cycloakylene-R27-, -R26-CH=CH-CH2- CH=CH-R27-, -R26-CH=CH-CH2-R47-R27-, -R26-R47-CH2-CH=CH-R27-, -R26-R47-CH2-R47-R27-, -R26- CH=CH-CD2-CH=CH-R27-, -R26-CH=CH-CD2-R47-R27-, -R26-R47-CD2-CH=CH-R27- or -R26-R47-CD2- R47-R27-; wherein, R47 for each occurrence, is an optionally substituted group independently selected from 3-10-membered carbocyclylene or 3-10-membered heterocyclylene; each double bond or cPr has a cis configuration;R65, R70, R78, R105 and R112 are each independently selected from: a bond, C1-C10 alkylene, C2-C10 alkenylene, C2-C10 alkynylene, Cs-Cg cycloalkylene, Cs-Cg cycloalkenylene, C5-Cio-spirocycloalkylene, C3- Cio-carbocyclylene, Cs-Cw-heterocyclylene, R26-CH=CH-C2-Csalkylene-CH=CH-R27-, -R26-CH=CH-C2- C6alkylene-R47-R27-, -R26-R47-C2-C6alkylene-CH=CH-R27-, -R26-R47-C2-C6alkylene-R47-R27-, - CH=CH-CH2-CH=CH-, -CH=CH-CH2-R47-, -R47-CH2-CH=CH-, -R47-CH2-R47-, -CH=CH-CD2-CH=CH- , - CH=CH-CD2-R47-, -R47-CD2-CH=CH- or -R47-CD2-R47-, wherein, R26 and R27 are each independently selected from: a bond, C1-C10 alkylene, C2-C10 alkenylene or C2-C10 alkynylene; R47 for each occurrence, is an optionally substituted group independently selected from 3-10-membered carbocyclylene or 3- 10-membered heterocyclylene;R66, R71, R79, R106 and R113 are each independently selected from: -H, -D, -CD3, linear or branched C1-C10 alkyl, C2-C 10 alkenyl, or C2-C10 alkynyl; or Cs-Cis-spirocycloalkylene-R28, Cs-Cis-carbocyclylene- R28, C3-Ci5-heterocyclylene-R28, -arylene-R28, or -heteroarylene-R28, -R26-R23, -R26-CH=CH-CH2- cPr-R28, -R26-cPr-CH2-CH=CH-R28, -R26-cPr-CH2-cPr-R28, R26-R149, -R26-R23, -R26-CH=CH-CH2- CH=CH-R28, -R26-CH=CH-CH2-R47-R28, -R26-R47-CH2-CH=CH-R28, -R26-R47-CH2-R47-R28, -R26- CH=CH-CD2-CH=CH-R28, -R26-CH=CH-CD2-R47-R28, -R26-R47-R47-R28, -R26-R47-R26-R47-R28, -R26- CD2-R47-R28, -R26-R47-CD2-R47-R28, -R26-R47-CD2-CH=CH-R28 or -R26-C3-C6cycloakylene-R26-C3-C6cycloakylene-R28;R123, R124, R125, R126, R127, R128, R132 and R133 are each independently is selected from: a bond, -O-, -OC(=O)-, -C(=O)O-, -OC(=O)O-, -S-C(=O)-, -C(=O)-S-, -S-S-, -C(=O)N(R164)-, -N(R164)C(=O)-, - N(R164)C(=O)N(R164)-, -O-C(=O)C(R164)2C(=O)O-, -C(=O)O-C(R164)2C(=O)O-, -O-C(=O)C(R164)2-O- C(=O)-, -O-C(R164)2C(=O)O-, -O-C(=O)C(R164)2C-O-, -O-C(R164)2-O-C(=O)-, -C(=O)-O-C(R164)2C-O-, -O- C(=O)-O-R191-O-C(=O)O-, -C(=O)-, -OC(=S)O-, -C(=S)O-,-OC(=S)- -C(=S)- -CH(R25)-, -C(R25)2-, - CH(OR25)-, -CH(NH[R25])-, -C(NH[OR25])-, -N(R25)-, -S-, -S(=O)-, -S(=O)2-, -OS(=O)2NH2, -S(=O)2NH2, - S(OH)-, -S(OH)2-, -C(=O)C(R25)2C(=O)-, -C(R25)2C(=O)-, -C(=O)C(R25)2-O-, -C(R25)2C(R25)2-, -C(R25)2-O- , -O-C(R25)2C(R25)2-, -O-C(R25)2C(=O)-, -C(=O)-O-R115-O-C(=O)-, -P(OH)(R25)-, -P(OR25)2-, -P(OR25)- and -P(=O)(OR25)-, -C(CI)2-C(=O)-O-, -O-C(=O)-C(CI)2-, -C(F)2-C(=O)-O-, -O-C(=O)-C(F)2-;R164, for each occurrence, is independently selected from: -H, -D, -CD3or optionally substituted, linear or branched C1-C6 alkyl;R191 is selected from: is a bond or optionally substituted, linear or branched C1-C6 alkylene, -C0-C3- alkylene-C3-Cio-carbocyclylene-Co-C3-alkylene- or -Co-C3-alkylene-C3-Cio-heterocyclylene-Co-C3- alkylene-; each R137 is independently selected from:ein each variable is defined above or for any aspect or embodiment herein.The structural formulas of an ionizable cationic lipid or cationic lipid of the present invention may be further defined for any aspect or embodiment as follows:In some embodiments, R1 or R19 is selected from:for each R1 or R19, one R33 is -R22-R21- (where R21, in the case of R19, is bound to R2) and each remaining R33 is independently selected from the group consisting of: -H, -D, -CH2(halo), -CH(halo)2, - C(halo)3, -F, -CF3, -Cl, -Br, -I, -CH3, -CD3, -CH2R25, -CH(R25)2, -C(R25)3, -OH, -OR25, -NH2, -NHR25, - N(R25)2, -SH, -SR25, -S(=O)H, -S(=O)R25, -S(=O)2H, -S(=O)2R25, -PH2, -PHR25, -P(R25)2, -P(OH)2, - POHR25, -P(R25)2, -P(OR25)2, -P(O)H2, -P(O)HR25, -P(O)(R25)2, -P(O)(OH)2, -P(O)(OR25)OH, - P(O)(OR25)2, -SiH3, -SiH2R25, -SiH(R25)2, and -Si(R25)3; and R21, R22, R25 and any remaining variables are as defined above or for any other embodiment herein.In some embodiments, R1 or R19 is selected from:wherein, each R33 is independently selected from the group consisting of: -H, -D, -CH2(halo), -CH(halo)2, - C(halo)3, -F, -CF3, -Cl, -Br, -I, -CH3, -CD3, -CH2R25, -CH(R25)2, -C(R25)3, -OH, -OR25, -NH2, -NHR25, - N(R25)2, -SH, -SR25, -S(=O)H, -S(=O)R25, -S(=O)2H, -S(=O)2R25, -PH2, -PHR25, -P(R25)2, -P(OH)2, - POHR25, -P(R25)2, -P(OR25)2, -P(O)H2, -P(O)HR25, -P(O)(R25)2, -P(O)(OH)2, -P(O)(OR25)OH, - P(O)(OR25)2, -SiH3, -SiH2R25, -SiH(R25)2, and -Si(R25)3; and R21, R22, R25 and any remaining variables are as defined above or for any other embodiment herein. In some further embodiments, R22 is selected from a bond, -CH2-, -[CH2]2-, -C(=O)- , -CH(OH)- or -NH-. In some further embodiments, R22 isselected from a bond, -CH2-, -[CH2]2-, -C(=O)- , -CH(OH)- or -NH- and R22 is selected from: a bond, - CH2-,-[CH2]2- or -C(=O)-.In some embodiments, R1 or R19 is selected from:wherein, for each Rl or R19, one R33 is -R22-R21- (where R21, in the case of Rl, is bound to R2) and each remaining R33 is independently selected from the group consisting of: -H, -D, -CH2(halo), -CH(halo)2, - C(halo)3, -F, -CF3, -Cl, -Br, -I, -CH3, -CD3, -CH2R25, -CH(R25)2, -C(R25)3, -OH, -OR25, -NH2, -NHR25, - N(R25)2, -SH, -SR25, -S(=O)H, -S(=O)R25, -S(=O)2H, -S(=O)2R25, -PH2, -PHR25, -P(R25)2, -P(OH)2, - POHR25, -P(R25)2, -P(OR25)2, -P(O)H2, -P(O)HR25, -P(O)(R25)2, -P(O)(OH)2, -P(O)(OR25)OH, - P(O)(OR25)2, -SiH3, -SiH2R25, -SiH(R25)2, and -Si(R25)3; and R21, R22, R25 and any remaining variables are as defined above or for any other embodiment herein.In some embodiments, Rl or R19 is selected from:for each R1 or R19, one R33 is -R22-R21- (where R21, in the case of Rl, is bound to R2) and each remaining R33 group is independently, selected from the group consisting of: -H, -D, -CH2(halo), - CH(halo)2, -C(halo)3, -F, -CF3, -Cl, -Br, -I, -CH3, -CD3, -CH2R25, -CH(R25)2, -C(R25)3, -OH, -OR25, -NH2, - NHR25, -N(R25)2, -SH, -SR25, -S(=O)H, -S(=O)R25, -S(=O)2H, -S(=O)2R25, -PH2, -PHR25, -P(R25)2, - P(OH)2, -POHR25, -P(R25)2, -P(OR25)2, -P(O)H2, -P(O)HR25, -P(O)(R25)2, -P(O)(OH)2, -P(O)(OR25)OH, -P(O)(OR25)2, -SiH3, -SiH2R25, -SiH(R25)2, and -Si(R25)3; and R21, R22, R25 and any remaining variables are as defined above or for any other embodiment herein.In some embodiments, Rl is selected from:; wherein, for each R1 or R19, one R33 is -R22-R21- (where R21, in the case of Rl, is bound to R2) and each remaining R33 group is independently selected from the group consisting of: -H, -D, -CH2(halo), - CH(halo)2, -C(halo)3,-F, -CF3, -Cl, -Br, -I, -CH3, -CD3, -CH2R25, -CH(R25)2, -C(R25)3, -OH, -OR25, -NH2, - NHR25, -N(R25)2, -SH, -SR25, -S(=O)H, -S(=O)R25, -S(=O)2H, -S(=O)2R25, , -S(=O)2NH2, -OS(=O)2NH2, - PH2, -PHR25, -P(R25)2, -P(OH)2, -POHR25, -P(R25)2, -P(OR25)2, -P(O)H2, -P(O)HR25, -P(O)(R25)2, - P(O)(OH)2, -P(O)(OR25)OH, -P(O)(OR25)2, -SiH3, -SiH2R25, -SiH(R25)2, and -Si(R25)3; and R21, R22, R25 and any remaining variables are as defined above or for any other embodiment herein. In some embodiments, one R33 is -R22-R21- and each remaining R33 group is independently selected from: - H, -D, -F, -CF3, -Cl, -Br, -I, -OH, -S(=O)2R25 or -NHR25. In some embodiments, one R33 is -R22-R21- and each remaining R33 group is independently selected from: -H, -D, -F, -Cl, -Br, or -OH.In some embodiments, R1 is selected from:wherein, each R33 is independently, selected from the group consisting of: -H, -D, -CH2(halo), - CH(halo)2, -C(halo)3, -F, -CF3, -Cl, -Br, -I, -CH3, -CD3, -OCH3, -OCD3, -CH2R25, -CH(R25)2, -C(R25)3, -OH, - OR25, -NH2, -NHR25, -N(R25)2, -SH, -SR25, -S(=O)H, -S(=O)R25, -S(=O)2H, -S(=O)2R25, -OS(=O)2NH2-, - S(=O)2NH2, -PH2, -PHR25, -P(R25)2, -P(OH)2, -POHR25, -P(R25)2, -P(OR25)2, -P(O)H2, -P(O)HR25, -P(O)(R25)2, -P(O)(OH)2, -P(O)(OR25)OH, -P(O)(OR25)2, -SiH3, -SiH2R25, -SiH(R25)2, and -Si(R25)3; and R21, R22, R25 and any remaining variables are as defined above or for any other embodiment herein.In some embodiments, R1 iswherein, each R33 is independently selected from: -H, -D, -F, -CF3, -Cl, -Br, -I, -OH, -S(=O)2CH3, -S(=O)2R25, -NHCH3 or -NHR25; and, R20, R21, R22 and R25 and any remaining variables are as defined above or for any other embodiment herein. In some further embodiments, R22 is selected from a bond, -CH2-, -[CFbh-, -C(=O)- -CH(OH)- or -NH-. In some further embodiments, R22 is selected from a bond, -CH2- or -C(=O)- and each R33 is independently selected from: -H, -D, -F, -Cl, -Br or -OH. In some further embodiments, R22 is selected from a bond, -CH2- or -C(=O)- and 1, 2, 3, 4 or 5 R33 groups is independently substituted with -OH, -F, Cl or Br, and the remaining R33 groups are -H or -D. In some further embodiments, R22 is selected from a bond, -CH2- or -C(=O)- and each R33 is -H or -D. In some further embodiments, of the 5 R33 groups: at least one R33 is -F; at least one R33 is -Cl; at least one R33 is -Br; at least two R33 are -F; at least two R33 are -Cl; at least two R33 are -Br; at least one R33 is -OH; at least two R33 are -OH; at least one R33 is -F -Cl or -Br and at least one R33 is -OH; at least two R33 is independently selected from -F, -Cl or -Br and at least one R33 is -OH; at least two R33 is -F and at least one R33 is -OH; at least two R33 are -Cl and at least one R33 is -OH; at least two R33 are -Br and at least one R33 is -OH; at least two R33 is independently selected from -F, -Cl or -Br; or at least 3 R33 are independently selected from -F, -Cl or -Br. In some embodiments, the R1 is bound to an R2 selected from: C1-C6 alkylene, C1-C6 alkenylene, C1-C6 alkynylene, -(CH2)O-B-0-(CH2)O-6-, -(CH2)O-B-0- (CH2)I-6-, (subscripts in the foregoing allowing a range of CH2), -(CH2)2-, -(CH2)3-, -(CH2)2-O-(CH2)3- or - CH2-O-(CH2)3-.In some embodiments, R1 iswherein,R22 is selected from a bond, -CH2-, -[CHzh-, -C(=O)- , -CH(OH)- or -NH-;R140 is R33 or R141; each R33 is independently selected from: -H, -D, -F, -CF3, -Cl, -Br, -I, -S(=O)2R25 or -NHR25;R141 isany remaining variables are as defined above or for any other embodiment herein. In some further embodiments, one R140 is R141 and each remaining R140 is independently R33. In some embodiments, the R1 is bound to an R2 selected frormC1-C6 alkylene, C1-C6 alkenylene, C1-C6 alkynylene, -(CH2)O-B-0-(CH2)O-6-, -(CH2)O-6-0-(CH2)I-6-, (subscripts in the foregoing allowing a range of CH2), -(CH2)2-, -(CH2)3-, -(CH2)2-O-(CH2)3- or -CH2-O- (CH2)3-.In some embodiments, Rl or R19 are selected from:; wherein, for each R1 or R19, one R33 is -R22-R21- (where R21, in the case of Rl, is bound to R2) and each remaining R33 group is independently, selected from the group consisting of: -H, -D, -CH2(halo), - CH(halo)2, -C(halo)3, -F, -CF3, -Cl, -Br, -I, -CH3, -CD3, -OCH3, -OCD3, -CH2R25, -CH(R25)2, -C(R25)3, -OH, - OR25, -NH2, -NHR25, -N(R25)2, -SH, -SR25, -S(=O)H, -S(=O)R25, -S(=O)2H, -S(=O)2R25, -OS(=O)2NH2-, - S(=O)2NH2, -PH2, -PHR25, -P(R25)2, -P(OH)2, -POHR25, -P(R25)2, -P(OR25)2, -P(O)H2, -P(O)HR25, - P(O)(R25)2, -P(O)(OH)2, -P(O)(OR25)OH, -P(O)(OR25)2, -SiH3, -SiH2R25, -SiH(R25)2, and -Si(R25)3; and R21, R22, R25 and any remaining variables are as defined above or for any other embodiment herein.A1through A5, A13and A14are each independently selected from: -CR33=, -C(R33)2-, -N=, -NR33-, -O-, -S- and -S(R46)-; for each R1 or R19, one R33 is -R22-R21- (where R21, in the case of Rl, is bound to R2) and each remaining R33 is independently selected from the group consisting of: a bond, -H, -D, -CH2(halo), - CH(halo)2, -C(halo)3, -F, -CF3, -Cl, -Br, -I, -CH3, -CD3, -OCH3, -OCD3, -CH2R25, -CH(R25)2, -C(R25)3, -OH, - OR25, -NH2, -NHR25, -N(R25)2, -SH, -SR25, -S(=O)H, -S(=O)R25, -S(=O)2H, -S(=O)2R25, -PH2, -PHR25, - P(R25)2, -P(OH)2, -POHR25, -P(R25)2, -P(OR25)2, -P(O)H2, -P(O)HR25, -P(O)(R25)2, -P(O)(OH)2, - P(O)(OR25)OH, -P(O)(OR25)2, -SiH3, -SiH2R25, -SiH(R25)2, and -Si(R25)3;R21, R22, R35, R46 and any remaining variables are as defined above or for any other embodiment herein;each L-2. is independently selected from a single or double bond, wherein selection of said single or double bond maintains proper valency, i.e., does not cause a valency violation such as a carbon, nitrogen or oxygen atom violating the octet rule. In some further embodiments, A1through A5, A13and A14are each independently selected from: -CR33=, -C(R33)j-, -N= and -NR33-; In some further embodiments, A1through A5, A13and A14are each independently selected from: -CR33= and -C(R33)j;In some embodiments, Rl or R19 is selected from:wherein, for each R1 or R19, one R33 is -R22-R21- (where R21, in the case of Rl, is bound to R2) and each remaining R33 is independently selected from the group consisting of: a bond, -H, -D, -CHjfhalo), -CH(halo)2, -C(halo)3, -F, -CF3, -Cl, -Br, -I, -CH3, -OCH3, -CD3, -OCD3, -CH2R25, -CH(R25)2, -C(R25)3, -OH, - OR25, -NH2, -NHR25, -N(R25)2, -SH, -SR25, -S(=O)H, -S(=O)R25, -S(=O)2H, -S(=O)2R25, -PH2, -PHR25, - P(R25)2, -P(OH)2, -POHR25, -P(R25)2, -P(OR25)2, -P(O)H2, -P(O)HR25, -P(O)(R25)2, -P(O)(OH)2, - P(O)(OR25)OH, -P(O)(OR25)2, -OS(=O)2NHR25-, -S(=O)2NHR25, -OS(=O)2NH2, -S(=O)2NH2, -SiH3, - SiH2R25, -SiH(R25)2, and -Si(R25)3;R21, R22, R25 and any remaining variables are as defined above or for any other embodiment herein; and each — is independently selected from a single or double bond, wherein selection of said single or double bond maintains proper valency, i.e., does not cause a valency violation such as a carbon, nitrogen or oxygen atom violating the octet rule.In some embodiments, R1 or R19 is selected from:wherein,A6through A11are each independently selected from: -CR33=, -C(R33)2-, -N=, -NR33-, -O-, -S-, -S(R46); for each R1 or R19 group, one R33 is -R22-R21- (where R21, in the case of Rl, is bound to R2) and each remaining R33 is independently selected from: a bond, -H, -D, -CH2(halo), -CH(halo)2, -C(halo)3, -F, - CF3, -Cl, -Br, -I, -CH3, -CD3, -OCD3, -CH2R25, -CH(R25)2, -C(R25)3, -OH, -OR25, -NH2, -NHR25, -N(R25)2, -SH, -SR25, -S(=O)H, -S(=O)R25, -S(=O)2H, -S(=O)2R25, -OS(=O)2NHR25, -S(=O)2NHR25, -OS(=O)2NH2, - S(=O)2NH2, -PH2, -PHR25, -P(R25)2, -P(OH)2, -POHR25, -P(R25)2, -P(OR25)2, -P(O)H2, -P(O)HR25, - P(O)(R25)2, -P(O)(OH)2, -P(O)(OR25)OH, -P(O)(OR25)2, -SiH3, -SiH2R25, -SiH(R25)2, or -Si(R25)3;R46 is selected from absent, =0 and (=0)2; R21, R22, R25 and any remaining variables are as defined above or for any other embodiment herein; andeach L-2. is independently selected from a single or double bond, wherein selection of said single or double bond maintains proper valency, i.e., does not cause a valency violation such as a carbon, nitrogen or oxygen atom violating the octet rule. In some further embodiments, A6through A11are each independently selected from: -CR33=, -C(R33)2-, -N= or -NR33-. In some further embodiments, A6through A11are each independently selected from: -CR33=, or -C(R33)2-.In some embodiments, R1 or R19 is selected from:or ; whereinA1through A14are each independently selected from: -CR33=, -C(R33)2-, -N=, -NR33-, -O-, -S-or - S(R46)-; for each R1 or R19, one R33 is -R22-R21- (where R21, in the case of Rl, is bound to R2) and each remaining R33 is independently selected from: a bond, -H, -D, -CH2(halo), -CH(halo)2, -C(halo)3, -F, - CF3, -Cl, -Br, -I, -CH3, -OCH3, -CD3, -OCD3, -CH2CH3, -CH2R25, -CH(R25)2, -C(R25)3, -OH, -OR25, -CN, -NO2, -NH2, -NHR25, -N(R25)2, -SH, -SR25, -S(=O)H, -S(=O)R25, -S(=O)CH3, -S(=O)2H, -S(=O)2R25, - OS(=O)2NHR25 -S(=O)2NHR25, -PH2, -PHR25, -P(R25)2, -P(OH)2, -POHR25, -P(R25)2, -P(OR25)2, -P(O)H2, -P(O)HR25, -P(O)(R25)2, -P(O)(OH)2, -P(O)(OR25)OH, -P(O)(OR25)2, -OS(=O)2NH2, -S(=O)2NH2, -SiH3, - SiH2R25, -SiH(R25)2, or -Si(R25)3;R21, R22, R25 and any remaining variables are as defined above or for any other embodiment herein;R46 is selected from: absent, =0 or (=0)2; andEach is independently selected from a single or double bond, wherein selection of said single or double bond maintains proper valency, i.e., does not cause a valency violation such as a carbon, nitrogen or oxygen atom violating the octet rule. In some embodiments, Rl or R19 is a bicyclic aromatic or hetero-aromatic group. In some embodiments, at least one variable selected from the group consisting of: A10, A11and A12, is independently selected from -CR33= or -C(R33)2-. In some embodiments, at least one variable selected from the group consisting of: A10, A11and A12, is independently selected from -N= or -NR33-.In some embodiments, R23 is an optionally substituted group selected from: a 3-18-membered saturated or partially unsaturated carboaryl, a 5-18-membered unsaturated carboaryl, a 3-18- membered saturated or partially unsaturated heteroaryl, or 5-18-membered unsaturated heteroaryl; wherein said heteroaryl contains one or more of heteroatoms independently selected from N, O or S as a ring atom. In some further embodiments, the carboaryl or heteroaryl is a 5-10 membered carboaryl or heteroaryl. In some further embodiments, the carboaryl or heteroaryl is a 5-10, 5-9, 5, 6 or 7 membered carboaryl or heteroaryl.In some embodiments, R23 is an optionally substituted group selected from: a 3-14-membered saturated or partially unsaturated carbocyclyl, a 5-14-membered unsaturated carbocyclyl, a 3-14- membered saturated or partially unsaturated heterocyclyl, or 5-14-membered unsaturatedheterocyclyl; wherein said heterocyclyl contains one or more of heteroatoms independently selected from N, O or S as a ring atom. In some further embodiments, R23 is independently substituted with at least one group selected from: -F, -Cl and -Br.In some embodiments, R23 is independently substituted with one or more R144, where R144 is selected from: -D, -CH3, -CD3, -OCD3, -CH2CH3,-CH2(halo), -CH(halo)2, -C(halo)3, -F, -CF3, -Cl, -Br, -I, -CH3, -OCH3, -CH2OCH3, -CH2R25, -CH(R25)2, -C(R25)3, -OH, -OR25, -CN, -NO2,-NH2, -NHR25, -N(R25)2, -SH, - SR25, -S(=O)H, -S(=O)R25, -S(=O)2H, -S(=O)2R25, S(=O)2CH3, -OS(=O)2NHR25 -S(=O)2NHR25, PH2, - PHR25, -P(R25)2, -P(OH)2, -POHR25, -P(R25)2, -P(OR25)2, -P(O)H2, -P(O)HR25, -P(O)(R25)2, -P(O)(OH)2, -P(O)(OR25)OH, -P(O)(OR25)2, -OS(=O)2NH2, -S(=O)2NH2, -SiH3, -SiH2R25, -SiH(R25)2, or -Si(R25)3; wherein R25 is as defined for any aspect or embodiment herein. In some further embodiments, each R144 is independently selected from: -H, -D, -F, -CF3, -Cl, -Br, -I, -OH, -S(=O)2CH3, -S(=O)2R25, - OS(=O)2NHR25, -S(=O)2NHR25, -NHCH3or -NHR25. In some further embodiments, each R144 is independently selected from: -F, -Cl, -Br, -OCH3or -OH. In some further embodiments, each R144 is independently selected from: -F, -Cl and -Br. In some further embodiments, the R23 is a 3-14- membered saturated or partially unsaturated carbocyclyl, a 5-14-membered unsaturated carbocyclyl, a 3-14-membered saturated or partially unsaturated heterocyclyl, or 5-14-membered unsaturated heterocyclyl; wherein said heterocyclyl contains one or more of heteroatoms independently selected from N, O or S as a ring atom. In some further embodiments, the carbocyclyl or heterocyclyl is idenpendently substituted with 1, 2, 3, 4 or 5 R144 groups. In some further embodiments, each R144 is independently selected from: -OH, -OCH3, -F, Cl or Br. In some further embodiments, at least one R144 is -F; at least one R144 is -Cl; at least one R144 is -Br; at least two R144 are -F; at least two R144 are -Cl; at least two R144 are -Br; at least one R144 is -OH; at least two R144 are -OH; at least one R144 is -OCH3; at least two R144 are -OCH3; at least one R144 is -F -Cl or -Br and at least one R144 is -OH or -OCH3; at least two R144 is independently selected from -F, -Cl or -Br and at least one R144 is -OH or - OCH3; at least two R144 is -F and at least one R144 is -OH or -OCH3; at least two R144 are -Cl and at least one R144 is -OH or -OCH3; at least two R144 are -Br and at least one R144 is -OH or -OCH3; at least two R144 is independently selected from -F, -Cl or -Br; or at least 3 R144 are independently selected from -F, -Cl or -Br.In some embodiments, R23 is an optionally substituted with R147-R143-. In some further embodiments, R147-R143- is an optionally substituted benzoyloxyphenyl-. In some embodiments, the benzoyloxyphenyl- is 4-(benzoyloxy)-phenyl. In some embodiments, the benzoyloxyphenyl- is substituted with one or more R144; wherein, R144 is independently selected from: -CH3, -CH2CH3,- CH2(halo), -CH(halo)2, -C(halo)3, -F, -CF3, -Cl, -Br, -I, -CH3, -OCH3, -CD3, -OCD3, or -CH2OCH3. In some further embodiments, R144 is independently selected from -F, -Cl, -Br, -OH or -OCH3, wherein R144 is independently substituted with at least one group selected from: -F, -Cl and -Br.In some embodiments, R23 is an optionally substituted group selected from: pyridinyl, pyridazinyl, pyrimidinyl, pyrazinyl, bicyclo(l.l.l)pentanyl, coumarinyl, furanyl, tetrahydrofuranyl, thiophenyl, pyrrolyl, pyrrolidinyl, pyranyl, piperidinyl, piperazinyl, imidazolyl, thiazolyl, 1,2-thiazinyl, 1,3-thiazinyl, 1,4-thiazinyl, dioxanyl, morpholinyl, 1,2-oxathiolanyl, 1,2-oxazolyl, 1,3-oxazolyl, isoxazolyl, silolyl, indolyl (2,3-benzopyrrolyl), isoindolyl, indolizinyl, quinolinyl, isoquinolinyl, carbazolyl, purinyl, aziridinyl, azetidinyl, pyrrolidinyl or piperidinyl.In some embodiments, R25 or R23 is an optionally substituted group selected from: 1-oxa-cyclobutan- 2-yl, tetrahydrofuran-3-yl, morpholin-4-yl, 2-thiacyclohex-l-yl, 2-oxo-2-thiacyclohex-l-yl, 2,2-dioxo-2-thiacyclohex-l-yl, 4-methyl-piperazin-2-yl, 2-hydroxy-aziridin-l-yl, 3-oxo-l-oxacyclobutan-2-yl, 2,2- dimethyl-tetrahydrofuran-3-yl, 3-carboxy-morpholin-4-yl, and l-cyclopropyl-4-methyl-piperazin-2-yl, oxetanyl, azetidinyl, thietanyl, tetrahydrofuranyl, dihydrofuranyl, tetrahydrothienyl, dihydrothienyl, imidazolidinyl, oxazolidinyl, imidazolinyl, isoxazolidinyl, pyrrolidinyl, 2-pyrrolinyl, 3-pyrroliny, tetrahydropyranyl, dihydropyranyl, dioxanyl, 1,3-dioxolanyl, 1,4-dithianyl, hexahydropyrimidine, morpholinyl, piperazinyl, piperidinyl, 2H-pyranyl, 4H-pyranyl, pyrazolidinyl, pyrazolinyl, 1, 2,3,6- tetrahydropyridinyl, tetrahydrothiopyranyl, l,l-dioxo-hexahydro-lX6-thiopyranyl, 1,1-dioxo-lX6- thiomorpholinyl, thiomorpholinyl, thioxanyl, and trithianyl.In some embodiments, R23 or R25 is an optionally substituted cycloalkyl. In some further embodiments, the optionally substituted cycloalkyl is selected from: cyclopropyl, cyclobutyl, cyclopentyl, cyclopentenyl, cyclohexyl, cyclohexenyl, cycloheptyl, cyclooctyl, cyclononyl, cyclodecyl and norbornyl.In some embodiments, R23 is an optionally substituted heteroaryl. In some further embodiments, R23 is an optionally substituted heteroaryl selected from: pyrrolyl, thienyl, thiazolyl, imidazolyl, furanyl, indolyl, isoindolyl, oxazolyl, isoxazolyl, benzothiazolyl, benzoxazolyl, quinolinyl, isoquinolinyl, pyridinyl, pyrimidinyl, pyrazinyl, pyridazinyl, triazolyl or tetrazolyl.In some further embodiments, R23 is an optionally substituted heteroaryl selected from: azepinyl, acridinyl, benzimidazolyl, benzindolyl, 1,3-benzodioxolyl, benzofuranyl, benzooxazolyl, benzo[d]thiazolyl, benzothiadiazolyl, benzo[b][l,4]dioxepinyl, benzo[b][l,4]oxazinyl, 1,4- benzodioxanyl, benzonaphthofuranyl, benzoxazolyl, benzodioxolyl, benzodioxinyl, benzoxazolyl, benzopyranyl, benzopyranonyl, benzofuranyl, benzofuranonyl, benzofurazanyl, benzothiazolyl, benzothienyl(benzothiophenyl), benzothieno[3,2-d]pyrimidinyl, benzotriazolyl, benzo[4,6]imidazo[l,2-a]pyridinyl, carbazolyl, cinnolinyl, cyclopenta[d]pyrimidinyl, 6,7-dihydro-5H- cyclopenta[4,5]thieno[2,3-d]pyrimidinyl, 5,6-dihydrobenzo[h]quinazolinyl, 5,6- dihydrobenzo[h]cinnolinyl, 6,7-dihydro-5H-benzo[6,7]cyclohepta[l,2-c]pyridazinyl, dibenzofuranyl, dibenzothiophenyl, furanyl, furazanyl, furanonyl, furo[3,2-c]pyridinyl, 5,6,7,8,9,10- hexahydrocycloocta[d]pyrimidinyl, 5,6,7,8,9,10-hexahydrocycloocta[d]pyridazinyl, 5,6,7,8,9,10- hexahydrocycloocta[d]pyridinyl, isothiazolyl, imidazolyl, indazolyl, indolyl, indazolyl, isoindolyl, indolinyl, isoindolinyl, isoquinolyl, indolizinyl, isoxazolyl, isoxazol-3-one, 5,8-methano-5,6,7,8- tetrahydroquinazolinyl, naphthyridinyl, 1,6-naphthyridinonyl, oxadiazolyl, 2-oxoazepinyl, oxazolyl, oxiranyl, 5,6,6a,7,8,9,10,10a-octahydrobenzo[h]quinazolinyl, 1-phenyl-TH-pyrrolyl, phenazinyl, phenothiazinyl, phenoxazinyl, phthalazinyl, pteridinyl, purinyl, pyranyl, pyrrolyl, pyrazolyl, pyrazolo[3,4-d]pyrimidinyl, pyridinyl, pyrido[3,2-d]pyrimidinyl, pyrido[3,4-d]pyrimidinyl, pyrazinyl, pyrimidinyl, pyridazinyl, pyrrolyl, quinazolinyl, quinoxalinyl, quinolinyl, isoquinolinyl, tetrahydroquinolinyl, 5,6,7,8-tetrahydroquinazolinyl, 5,6,7,8-tetrahydrobenzo[4,5]thieno[2,3- d]pyrimidinyl, 6,7,8,9-tetrahydro-5H-cyclohepta[4,5]thieno[2,3-d]pyrimidinyl, 5, 6,7,8- tetrahydropyrido[4,5-c]pyridazinyl, thiazolyl, thiadiazolyl, thiapyranyl, triazolyl, tetrazolyl, triazinyl, thieno[2,3-d]pyrimidinyl, thieno[3,2-d]pyrimidinyl, thieno[2,3-c]pyridinyl or thiophenyl (i.e. thienyl).In some embodiments, R23 is selected from:wherein:R35 and R41 are each independently selected from:R36 through R40 and R42 through R45 are each independently selected from -CR24=, -C(R24)2-, -N=, - NR24-, -O-, -S- or -S(R46)-;R24 and R60 is as defined above or for any aspect or embodiment herein;R46 is selected from: absent (i.e., two lone electron pairs), =0 or (=O)2; each L-L is independently selected from a single or double bond, wherein selection of said single or double bond maintains proper valency, i.e., does not cause a valency violation such as a carbon, nitrogen or oxygen atom exceeding the octet valency rule for the period having C, N and O.In some further embodiments, R35 is -CR24- or -CH(R24)-. In some further embodiments, the 6- membered ring formed by R35 through R40 or the 5-membered ring formed by R41 through R45 is an aryl or heteroaryl group.In some embodiments, R23 is selected from:; whereinR35 and R41 are each independently selected from:one or two substituents selected from R36 through R40 and one or two substituents selected from R42 through R45 are each independently selected from -N= or -NR24-; and the remaining substituents selected from R36 through R40 and R42 through R45 are selected from -CR24= or -C(R24)2-; whereineach L-2. is independently selected from a single or double bond, wherein selection of said single or double bond maintains proper valency, i.e., does not cause a valency violation such as a carbon, nitrogen or oxygen atom violating the octet rule. In some further embodiments, the 6-membered ring formed by R35 through R40 or the 5-membered ring formed by R41 through R45 is an aryl or heteroaryl.In some embodiments, R23 is selected from:In some embodiments, R25 for each occurrence, is independently selected from: -H, -D or an optionally substituted group selected from: Ci-Cio alkyl, C2-C10 alkenyl, C2-C10 alkynyl, 3-14-membered cycloalkyl, 5-14-membered spirocycloalkyl or 3-14-membered heterocycloalkyl. In some further embodiments, R25 for each occurrence, is independently selected from: 3-10-membered cycloalkyl, 5-10-membered spirocycloalkyl or 3-10-membered heterocycloalkyl.In some embodiments, R25, for each occurrence, is independently selected from: -CH3, -CD3, -CF3, - CH2RII4, -CH(R114)2, -CH2(halo), -CH(halo)2, -C(halo)3, -OH, -NH2, -NHR114, -SH, -S(=O)H, -S(=O)2H, - PH2, -PHR114, -P(OH)2, POHR114, -P(OH)R114, -P(O)H2, -P(O)HR114, -P(O)(OH)2, -P(O)(OR114)OH, - P(O)(OR114)2, -SiH3, -SiH2R114 or -SiH(R114)2; wherein R114 is as defined above or for any other embodiment herein.In some embodiments, R114, for each occurrence, is independently selected from: -H, -D, -CD3or an optionally substituted, linear or branched, C1-C6 alkyl, C3-Cg alkenyl or C3-Cg alkynyl. In some embodiments, R114, for each occurrence, is independently selected from an optionally substituted, linear or branched: C3-C3alkyl, C3-Cg alkyl, C3-Cg alkenyl or C3-Cg alkynal.In some embodiments, R25, for each occurrence, is independently selected from: -H, -D, -CD3, -CH3, - CH2CH3, -F, -Cl, -Br, -I, -OCH3, -OCD3, -OH or -NH2. In some embodiments of R25, for each occurrence, the carbocyclyl or heterocyclyl is independently selected from a 5-10 membered carbocyclyl or heterocyclyl. In some further embodiments, the carbocyclyl or heterocyclyl is a 5-10, 5-9, 5, 6 or 7 membered carbocyclyl or heterocyclyl. In some further embodiments of R25, the heterocyclyl contains one or more heteroatoms independently selected from N or O as a ring atom. In some further embodiments, of R25, each ring heteroatom of the heterocyclyl is either N or O.In some embodiments, R25 is an optionally substituted C1-C10 alkyl, C2-C10 alkenyl or C2-C10 alkynyl that is linear and in other embodiments, R25 is a C1-C10 alkyl, C2-C10 alkenyl or C2-C10 alkynyl that is branched.In some embodiments, R25, is a C1-C10 alkyl, C2-C10 alkenyl, C2-C10 alkynyl, C5-C16 spirocycloalkyl, C5-C16- cycloalkyl or C1-C10 heterocycloalkyl that is substituted with one or more substituents, each independently, selected from: -CH2(halo), -CH(halo)2, -C(halo)3, -F, -Cl, -Br, -CF3, -I, -CH3, -OCH3, -CD3, - OCD3, -OH, -NH2, -SH, -S(=O)H, -S(=O)2H, -PH2, -P(OH)2, -P(O)H2, -P(O)(OH)2, -SiH3or R15.In some embodiments, R25 is an optionally substituted group selected from: spiropentanyl, spriohexanyl, spiroheptanyl, spirooctanyl, spirononanyl, or spirodecanyl.In some embodiments, R22 is selected from: -CH2-, -[CH2]2-, -[CH2]3-, isopropylene, butylene, isobutylene, -N(R116)-, -C(R145)2-, NHC(=O)O-, -OC(=O)NH-, -P(OH)- or -P(O)OH-, wherein, R116 is -H, -D, -CD3, or C1-3 alkyl, and R145, for each occurrence, is independently selected from: -H, -D, -OCH3, - OH, -CD3, -OCD3, or C1-C6 alkyl;. In some embodiments, R145 is selected from: -H, -D, -CD3, -OCD3, methoxy, -OH, methyl, ethyl or propyl. In some embodiments, R22 is selected from: -CH2-, -[CH2]2-, - NH-,In some embodiments of R24, halo is independently selected from: -F, -Cl or -Br. In some embodiments, R24 is selected from: -OH, -CH3, -CD3, -OCD3, -CH2R25, -CH(R25)2, -NH2, -NHR25, -SH, - S(=O)H, -S(=O)2H, -PH2, -PHR25, -P(O)(OH)2, -P(OH)2, -POHR25, -P(O)H2, -P(O)(OR25)OH, -P(O)HR25, - SiHs, -SiH2R25 or -SiH(R25)2; wherein R25 is as defined above or for any other embodiment herein. In some embodiments, R24 is an optionally substituted, linear or branched group selected from: Ci-C8- alkyl, C2-C8 alkenyl, C2-C8alkynyl, C1-C6- alkyl, C2-C6 alkenyl, C2-C6 alkynyl, C1-C4- alkyl, C2-C4alkenyl or C2-C4alkynyl.In some embodiments, R2 is selected from: a bond, a linear, branched or optionally substituted group selected from: -[CH2]O-3CH=CH[CH2]I.3-, -[CH2]0-3CH=CHCH2-, -[CH2]I.3CH=CH[CH2]I-3-, -[CH2]0- 3CEC[CH2]I-3-, -[CH2]I-3CEC[CH2]I-3-, -Co-Cg acyclic hydrocarbylene-CH(R47)-Co-Cg akylene-, -Co-Cg acyclic hydrocarbylene-CH(R47)-[CH2]o-3CH=CH[CH2]i-3, -Co-Cg acyclic hydrocarbylene-CH(R47)-[CH2]o- 3CHECH[CH2]I-3-, -Co-Cg akylene-R47-Co-Cg akylene-, -Co-Cg akylene-R146-Co-Cg akylene-, -Co-Cg acyclic hydrocarbylene-R143-Co-Cg akylene-, -Co-Cg acyclic hydrocarbylene-R143-[CH2]o-3CH=CH[CH2]i-3, -Co-Cg acyclic hydrocarbylene-R143-[CH2]o-3C=C[CH2]i-3-, -Co-Cg akylene-R143-Co-Cg akylene-, -Co-Cg acyclic hydrocarbylene-R146-Co-Cg akylene-, -Co-Cg acyclic hydrocarbylene-R146-[CH2]o-3CH=CH[CH2]i-3, -Co-Cg acyclic hydrocarbylene-R146-[CH2]o-3C=C[CH2]i-3-, -Co-Cg akylene-R146-Co-Cg akylene-, -Co-Cg acyclic hydrocarbylene-CH(R147)-Co-Cg akylene-, -Co-Cg acyclic hydrocarbylene-CH(R147)-[CH2]o-3CH=CH[CH2]I-3, -Co-Cg acyclic hydrocarbylene-CH(R147)-[CH2]o-3C=C[CH2]i-3-, -C0-C8akylene-R147-C0- Cg akylene-, Co-Cg acyclic hydrocarbylene-O-C1-C6 acyclic hydrocarbylene, -Co-Cg acyclic hydrocarbylene-O-Co-Cg akylene-, -Co-Cg acyclic hydrocarbylene-O-C1-C6 akylene-, -Co-Cg acyclic hydrocarbylene-0-[CH2]o-3CH=CH[CH2]i-3, -Co-Cg acyclic hydrocarbylene-0-[CH2]o-3C=C[CH2]i-3-, -Co-Cg akylene-O-Co-Cg akylene-, -C0-C8akylene-O-C1-C6 akylene- or -(CH2)o-6-0-(CH2)0-6-; wherein R47, R143, R146 and R147 are as defined above or for any other embodiment herein.In some embodiments of R2, the Co-Cg acyclic hydrocarbylene is a C2-C8acyclic hydrocarbylene, C2-C6 acyclic hydrocarbylene or C1-C3 acyclic hydrocarbylene.In some embodiments, R2 is a linear, branched or optionally substituted group selected from: Ci-Cg alkylene, C2-C8alkenylene, C2-C8alkynylene, C2-C8internal alkenylene, C2-C8internal alkynylene, -Ci- C6akylene-R47-C1-C6 akylene-, -C1-C6 akylene-R143-C1-C6 akylene-, -C1-C6 akylene-R146-C1-C6 akylene-, -C1-C6 akylene-CH(R147)-C1-C6 akylene-, -C1-C6 akylene-O-C1-C6 akylene-, -C1-C6 akylene-S- C1-C6 akylene-, -C1-C6 akylene-S-S-C1-C6 akylene-, -C1-C6 akylene-N(R114)-Ci-Cs akylene-; wherein R47, R114, R143, R146 and R147 are as defined above or for any other embodiment herein. In some further embodiments, alkylene, alkenylene or alkynylene is linear. In some further embodiments, the alkylene, alkenylene or alkynylene is unsubstituted. In some further embodiments, the alkylene, alkenylene or alkynylene is linear and unsubstituted.In some embodiments, R2 is a linear, branched or optionally substituted group selected from: C1-C6 alkylene, C1-C6 alkenylene, C1-C6 alkynylene, C1-C6 internal alkenylene, C1-C6 internal alkynylene, -C1-C3 alkylene-R47-C1-C3 alkylene-, -C1-C3 alkylene-R143-C1-C3 alkylene-, -C1-C3 alkylene-R146-C1-C3 alkylene- , -C1-C3 alkylene-CH(R147)-C1-C3 alkylene-, -C1-C3 alkylene-O-C1-C3 alkylene-, -C1-C3 alkylene-S-Ci-C3alkylene-, -C1-C3 alkylene-S-S-Ci-Cs alkylene-, -C1-C3 alkylene-N(R114)-C1-C3 alkylene-; wherein R47, R114, R143, R146 and R147 are as defined above or for any other embodiment herein. In some further embodiments, alkylene, alkenylene or alkynylene is linear. In some further embodiments, the alkylene, alkenylene or alkynylene is unsubstituted. In some further embodiments, the alkylene, alkenylene or alkynylene is linear and unsubstituted. In some further embodiments, R47 is a Cs-Cg heterocyclylene or C3-C8carbocyclylene.In some embodiments, R2 is selected frormC1-C6 alkylene, C1-C6 alkenylene, C1-C6 alkynylene, -(CH2)o-6-0-(CH2)O-6-, -(CH2)O-6-0-(CH2)I.6-, -(CH2)2-, -(CH2)3-, -(CH2)2-O-(CH2)3- or -CH2-O-(CH2)3-.In some embodiments, R2 is selected from: -[CH2]o-6-0-[CH2]o-6- or -[CH2]O-6-0-[CH2]I.6-, -[CH2]I.6-O- [CH2]O-6- , -l,4-piperazin-l,4-diyl-, -l,3-piperazin-l,3-diyl- or -l,4-diazepan-l,4-diyl.In some embodiments, R2 or R146 is selected from: C3-C7 cycloakylene or C3-C7 cycloakenylene. In some embodiments, R2 or R146 is cyclopropylene, cyclobutylene, cyclopentylene, cyclohexylene, cyclopropendiyl, cyclobutendiyl, cyclopentendiyl or cyclohexendiyl.In some embodiments, R2 is R29-O-R30 or R29-R47-R30, wherein R29 and R30 are each independently an optionally substituted, straight or branched C1-C6 alkylene, C1-C6 alkenylene, C1-C6 alkynylene, and R47 is defined above or for any aspect / embodiment herein. In some embodiments, R29 and R30 are each independently an optionally substituted, straight or branched C1-C6 alkylene. In some further embodiments, R29 and R30 are different. In some embodiments, R29 and R30 are each independently -[CH2]O-6-. In some embodiments, R29 is -CH2- and R30 is -[CH2]2_6-. In some embodiments, R29 is - [CH2]2- and R30 is -CH2- or -[CH2]3-6-. In some embodiments, R29 is -[CH2]3- and R30 is -[CH2]I.2- or - [CH2]4-6-- In some embodiments, R29 is -[CH2]4- and R30 is -[CH2]I-3- or -[CH2]5-6-. In some embodiments, R29 is -[CH2]4- and R30 is -[CH2]I-3- or -[CH2]5-6-. In some embodiments, R29 is -[CH2]5- and R30 is - [CH2]I.4- or -[CH2]6-. In some embodiments, R29 is -[CH2]g- and R30 is -[CH2]I_5-. In some embodiments, R2 is selected frormC1-C6 alkylene, C1-C6 alkenylene, Ci-Cg alkynylene, -[CH2]o-6-0-[CH2]o-6-, -[CH2]o-6-0- [CH2]I-6-, -[CH2]2-, -[CH2]3-, -[CH2]2-O-[CH2]3- or -CH2-O-[CH2]3-. In some embodiments, R2 is -0-[CH2]O-B- . In some embodiments, R2 is -O-[CH2]I.6-. In some embodiments, R2 is -O-[CH2]2.6-. In some embodiments, R2 is -[CH2]2-, -[CH2]3-, -[CH2]2-O-[CH2]3- or -CH2-O-[CH2]3-. In some embodiments, R29 is -[CH2]2.6- and R30 is -CH2-. In some embodiments, R29 is -CH2- or -[CH2]3-6- and R30 is -[CH2]2-. In some embodiments, R29 is -[CH2]I.2- or -[CH2]4.6- and R30 is -[CH2]3-. In some embodiments, R29 is - [CH2]I-3- or -[CH2]5-6-. and R30 is -[CH2]4-. In some embodiments, R29 is -[CH2]I.4- or -[CH2]g and -R30 is -[CH2]5-. In some embodiments, R29 is -[CH2]I_5- and R30 is -[CH2]g-. In some further embodiments, R29 and R30 are each independently selected from: methylene, ethylene, propylene, isopropylene, butylene, isobutylene, tert-butylene, pentylene, 1,1-dimethylpropylene, 2,2-dimethylpropylene, 1- methylbutylene, 2-methylbutylene, 3-methylbutylene, 1-ethylpropylene, 1,2-dimethylpropylene or hexylene.In some embodiments of R47, for each occurrence, is independently a 3-10-membered group independently selected from: cycloalkylene, cycloalkenylene, saturated heterocyclene or partially unsaturated heterocyclene. In some embodiments, R47, for each occurrence,' is independently selected from: an optionally substituted 3-8-membered saturated, partially unsaturated orunsaturated carbocyclene or an optionally substituted 3-8-membered saturated, partially unsaturated or unsaturated heterocyclene. In some further embodiments, R47, for each occurrence, is independently a 3-7-membered group independently selected from: cycloalkylene, cycloalkenylene, saturated heterocyclene or partially unsaturated heterocyclene. In some further embodiments, R47, for each occurrence, is independently selected from: cPr, cBu or cHx.In some embodiments, R147 is selected from C3-C7 cycloakyl or C3-C7 heterocycloakyl. In some embodiments, R147 is cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, cyclopropenyl, cyclobutenyl, cyclopentenyl, cyclohexenyl, 1,3-cyclohexadienyl or 1,4-cyclohexadienyl.In some embodiments, R143 is selected from: -NH-, NH-C(=O)-O- or -O-C(=O)-NH-,In some embodiments, each R50 is independently selected from:-R62-R123-R63-R66,-R62-R123-R63-R74-R66,-R62-R123-R63-R124-R64-R65-R66,-R62-R123-R63-R124-R64-R66,-R62-R123-R63-R124-R64-R74-R66,-R62-R123-R63-R124-R64-R74-R127-R66,-R62-R123-R63-R124-R64-R74-R127-R75-R66,-R62-R123-R63-R124-R64-R74-R127-R75-R65-R66 or-R67-R125-R68-R126-R69-R70-R71; wherein said 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 further embodiments, an R50 or R137 has a longest linear chain (i.e., excluding side groups) with a total number of carbon atoms that is selected from: 16, 17, 18, 19, 20, 21 or 22. In some further embodiments, an R50 or R137 has a longest linear chain (i.e., excluding side groups) with a total number of atoms that is selected from: 16, 17, 18, 19 or 20. In some further embodiments, an R50 or R137 has a longest linear chain (i.e., excluding side groups) with a total number of atoms that is selected from: 18, 19, 20, 21 or 22 carbon atoms.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, R3, R4, Rll and R12 are each independently selected from: a bond, -R26-C1- C10 alkylene-R27-, -R26-C2-CIO alkenylene-R27-, -R26-C2-CIO alkynylene-R27-, -R26-C3-C10 carbocyclylene-R27-, -R26-C3-C10 heterocyclylene-R27-; wherein, R26 and R27 are each independently selected from: a bond, or an optionally substituted C1-C6 alkylene, C2-C6alkenylene or C2-C6alkynylene. In some further embodiments, R26 and R27 are each independently selected from: a bond, or an optionally substituted C1-C3 alkylene, C2-Cs alkenylene or C2-Cs alkynylene.In some embodiments, R3, R4, Rll and R12 are each independently selected from: C1-C10 alkylene, C2-C10 alkenylene or C2-C10 alkynylene, C3-C10 carbocyclylene or C3-C10 heterocyclylene.In some embodiments, R3, R4, Rll and R12 are each independently selected from: a bond, C1-C10 alkylene, C2-C10 alkenylene or C2-C10 alkynylene. In some embodiments, R3, R4, Rll and R12 are each independently selected from:, R26-C3-C6-cycloalkylene-R27-, -R26-CH=CH-CH2-CH=CH-R27- , -R26- CH=CH-CH2-R47-R27-, -R26-R47-CH2-CH=CH-R27-, -R26-R47-CH2-R47-R27-, -R26-CH=CH-CD2-CH=CH- R27-, -R26-CH=CH-CD2-R47-R27-, -R26-R47-CD2-CH=CH-R27-, -R26-R47-CD2-R47-R27-, -R26-CH=CH- CH2-cPr-R27-, -R26-cPr-CH2-CH=CH-R27-, -R26-cPr-CH2-cPr-R27-,-R26-CH=CH-CD2-cPr-R27-, -R26-cPr- CD2-CH=CH-R27- or -R26-cPr-CD2-cPr-R27-; wherein, R26 and R27, for each occurrence, is independently selected from: a bond, C1-C10 alkylene, C1-C10 alkenylene or C1-C10 alkynylene; and R47, for each occurrence, is independently Cs-Cg-cycloalkylene. In some further embodiments, each cPr has a cis configuration. In some further embodiments, each double bond has a cis configuration. In some further embodiments, R26 and R27 are each independently selected from: a bond, C1-C6 alkylene, C2- Cg alkenylene or C2-C6 alkynylene. In some further embodiments, R26 and R27 are each independently selected from: a bond, C1-C3 alkylene, C2-C3 alkenylene or C2-C3 alkynylene.In some embodiments, R5, R6, R9 and RIO are each independently selected from the group consisting of: -O-CH2-, -O-CH(OH)2-, -N(OH)-, -NH- or -C(NH[OH])-.In some embodiments, R7 and R8 are each independently selected from: a bond, R3, -CH(OH)-, - CH(NH2)-, -C(NH[OH])-, -NH-, -N(OH)-, -C(=O)-O-R115-O-C(=O)-, -P(OH)-, -P(OH)(R25)-, -P(=O)(OH)-, - P(OR25)2-, -P(OR25)- or -P(=O)(OR25)-; wherein, R25 and R115 are as defined above or for any aspect or embodiment herein. In some embodiments, R7 and R8 are each independently selected from R3.In some embodiments, R115, for each occurrence, is each independently an optionally substituted group selected from: linear or branched Ci-Cg alkylene, C2-C8 alkenylene or C2-C8 alkynylene; -R26-C3- C10 heterocyclylene-R27- or -R26-C3-C10 carbocyclylene-R27-; -R26-C3-Cg-carbocyclylene-R27-, -R26-C3- C6-heterocyclylene-R27-; wherein, R26 and R27 are defined above or for any aspect or embodiment herein. In some embodiments, R115, for each occurrence, is each independently an optionally substituted group selected from: C3-C10 heterocyclylene, C3-C10 carbocyclylene; or -Cs-Cg- carbocyclylene-, -Cs-Cg-heterocyclylene-, -R26-C3-Cg-cycloakylene-R27- -R26-CH=CH-CH2-CH=CH-R27- , -R26-CH=CH-CH2-cPr-R27-, -R26-cPr-CH2-CH=CH-R27-, -R26-cPr-CH2-cPr-R27-, -R26-CH=CH-CD2- CH=CH-R27-, -R26-CH=CH-CD2-cPr-R27-, -R26-cPr-CD2-CH=CH-R27- or -R26-cPr-CD2-cPr-R27-; wherein, R26 and R27 are defined above or for any aspect or embodiment herein. In some embodiments, R115, for each occurrence, are each independently an optionally substituted group selected from: linear or branched C1-C6 alkylene, C2-C6 alkenylene or C2-C6 alkynylene; 3-7 membered heterocyclylene or 3-7 membered carbocyclylene; or -R26-C3-Cg-carbocyclylene-R27-, -R26-C3-Cg- heterocyclylene-R27-, -R26-CH=CH-CH2-CH=CH-R27- , -R26-CH=CH-CH2-cPr-R27-, -R26-cPr-CH2- CH=CH-R27-, -R26-cPr-CH2-cPr-R27-, -R26-CH=CH-CD2-CH=CH-R27-, -R26-CH=CH-CD2-cPr-R27-, -R26- cPr-CD2-CH=CH-R27- or -R26-cPr-CD2-cPr-R27-; wherein, R26 and R27 are defined above or for any aspect or embodiment herein. In some embodiments, R115 is -R26-C3-Cg-cycloakylene-R27-; wherein R26 and R27 are defined above or for any aspect or embodiment herein. In some embodiments of R115, R26 and R27 are each independently selected from: a bond, C1-C3 alkylene, C2-C3 alkenylene or C2-C3 alkynylene. In some embodiments of R115, each double bond or cPr has a cis configuration.In some embodiments, R15, R16, R17 and R18 are each independently selected from: -H, -D, -CD3; an optionally substituted linear or branched, C1-C10 alkyl, C2-C10 alkenyl, or C2-C10 alkynyl; an optionally substituted, C3-C7 cycloalkyl, C3-C7 cycloalkenyl, C5-Cio-spirocycloalkyl, Ca-Cw-carbocyclyl, C3-C10- heterocyclyl, aryl, or heteroaryl; or -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-cPr-CD2-cPr-R28, -R26-CD2-cPr-R28, -R26-cPr-CD2-cPr-R28, -R26-cPr-CD2-cPr-R28, -R26-cPr-CD2-CH=CH-R28, -R26-cPr- CD2-cPr-R28; or -R26-C3-C6cycloakylene-R26-C3-C6cycloakylene-R28; wherein, each R26 is independently as defined above or for any other embodiment herein, and R28 is selected from: -H, - D, -CD3, C1-C10 alkyl, C2-C10 alkenyl or C2-C10 alkynyl. In some embodiments or R15, R16, R17 and R18, each double bond or -cPr- has a cis configuration. In some embodiments, R15, R16, R17 and R18 are each independently selected from: -H, -D -CD3; an optionally substituted linear or branched, C1-C6 alkyl, C2-C6alkenyl, or C2-C6alkynyl; an optionally substituted, C3-C7cycloalkyl, C3-C7cycloalkenyl, C5- Cio-spirocycloalkyl, C3-C7-carbocyclyl, C3-C7-heterocyclyl, phenyl, or 5-7 membered heteroaryl. In some further embodiments, each R26 is independently selected from: a bond or linear or branched C1-C6 alkylene, C1-C6 alkenylene or C1-C6 alkynylene, and R28 is selected from: -H, C1-C6 alkyl, C2-C6 alkenyl or C2-C6 alkynyl. In some further embodiments, each R26 is independently selected from: a bond, Ci- C3 alkylene, C2-C3 alkenylene or C2-C3 alkynylene. In some further embodiments, R28 is selected from: -H, C1-C3 alkyl, C2-C4 alkenyl or C2-C4 alkynyl.In some embodiments, R62, R63, R64, R67, R68, R69, R74, R75, R76, R77, R108, R109, R110 and Rill are each independently selected from: a bond, C1-C6 alkylene, C2-C6 alkenylene, C2-C6 alkynylene, C3- Cg cycloalkylene, C3-Cg cycloalkenylene, C3-C7-carbocyclylene, C3-C7-heterocyclylene, -R26-CH=CH-C2- C4alkylene-CH=CH-R27-, -R26-CH=CH-C2-C6 alkylene-R47-R27-, -R26-R47-C2-C6alkylene-CH=CH-R27-, - R26-R47-C2-C6 alkylene-R47-R27-, -R26-C3-Cg-cycloakylene-R27-, -R26-CH=CH-CH2-CH=CH-R27-, -R26- CH=CH-CH2-R47-R27-, -R26-R47-CH2-CH=CH-R27-, -R26-R47-CH2-R47-R27-, -R26-CH=CH-CD2-CH=CH- R27-, -R26-CH=CH-CD2-R47-R27-, -R26-R47-CD2-CH=CH-R27- or -R26-R47-CD2-R47-R27-; R26 and R27 are each independently selected from: a bond, C1-C10 alkylene, C2-C10 alkenylene or C2-C10 alkynylene; R47, for each occurrence, is independently a 3-10-membered carbocyclylene or a 3-10-membered heterocyclylene; and each double bond or cPr has a cis configuration.In some embodiments, R65, R70, R78, R105 and R112 are each independently selected from: a bond, C1-C6 alkylene, C2-Cg alkenylene, C2-Cg alkynylene, C3-Cg cycloalkylene, C3-Cg cycloalkenylene, C5-C10- spirocycloalkylene, 3-7 membered carbocyclylene, 3-7 membered heterocyclylene, R26-CH=CH-C2- C4alkylene-CH=CH-R27-, -R26-CH=CH-C2-C4alkylene-R47-R27-, -R26-R47-C2-C6alkylene-CH=CH-R27-, - R26-R47-C2-C6alkylene-R47-R27-, -CH=CH-CH2-CH=CH-, -CH=CH-CH2-R47-, -R47-CH2-CH=CH-, -R47- CH2-R47-, -CH=CH-CD2-CH=CH-, -CH=CH-CD2-R47-, -R47-CD2-CH=CH- or -R47-CD2-R47-; wherein, R47 for each occurrence, is an optionally substituted group independently selected from 3-7-membered carbocyclylene or 3-7-membered heterocyclylene. In some embodiments of R65, R70, R78, R105 and R112, each double bond has a cis configuration. In some embodiments, R65, R70, R78, R105 and R112 are each independently selected from: -R26-CH=CH-C2-C4alkylene-cPr-R27-, -R26-cPr-C2-C6alkylene- CH=CH-R27-, -R26-cPr-C2-C6alkylene-cPr-R27-, -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-, wherein, each double bond or cPr has a cis configuration.In some embodiments, R66, R71, R79, R106 and R113 are each independently selected from: -H, linear or branched Ci-C8alkyl, C2-C8alkenyl, or C2-C8alkynyl, C5-Ci0-spirocycloalkyl, C3-Ci0-carbocyclyl, C3-Ci0- heterocyclyl, 5-10 membered aryl or 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-R28; or -R26-C3-Cg cycloakylene-R26-C3-Cs cycloakylene-R28; wherein, R26 is as defined above or for any other embodiment herein, and R28 is selected from: -H, C1-C10 alkyl, C2-Ci0alkenyl or C2-Ci0alkynyl. In some further embodiments of R66, R71, R79, R106 and R113, each double bond or -cPr- has a cis configuration. In some further embodiments, each R26 is independently selected from: a bond or linear or branched C1-C6 alkylene, C1-C6 alkenylene or C1-C6 alkynylene, and R28 is selected from: -H, C1-C6 alkyl, C2-C6 alkenyl or C2-C6 alkynyl. In some further embodiments, each R26 is independently selected from: a bond, C1-C3 alkylene, C2-C3alkenylene or C2-C3alkynylene. In some further embodiments, R28 is selected from: -H, C1-C3 alkyl, C2-C4alkenyl or C2-C4alkynyl.In some embodiments, R149 is:In some embodiments, R123, R124, R125, R126, R127, R128, R132 and R133 are each independently is selected from: In some embodiments, R123, R124, R125, R126, R127, R128, R132 and R133 is selected from: -CH(OH)-, -CH(NH2)-, C(NH[OH])-, -P(OH)-, or -P(=O)(OH)-. In some embodiments of R123, R124, R125, R126, R127, R128, R132 and R133, R164, for each occurrence, is independently selected from: -H or optionally substituted, linear or branched C1-C3 alkyl; and R191 is selected from: is a bond or optionally substituted, linear or branched C1-C3 alkylene, -Co-C3-alkylene-C3-C7- carbocyclylene-Co-C3-alkylene- or -Co-C3-alkylene-C3-C7-heterocyclylene-Co-C3-alkylene-.In some embodiments for one or more of R24, R25, R33 or R114, each of said halo is independently selected from -F, -Cl or -Br, and in further embodiments, -Br or -Cl.In some embodiments for any one or more variable selected from R15, R16, R17, R18, R23, R25, R66, R71, R79, R106 or R113, 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 for any one or more variable selected from R15, R16, R17, R18, R23, R25, R66, R71, R79, R106 or R113, said carbocyclyl or heterocyclyl is independently an optionally substituted group selected from: lH-pyrrolizidinyl, 1,2-dihydroquinolinyl, 1,5-naphthyridinyl, 1,8-naphthyridinyl,IH-indazolyl, lH-isochromenyl, IH-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 for any one or more variable selected from R15, R16, R17, R18, R23, R25, R66, R71, R79, R106 or R113, 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, 1,2- thiazinyl, 1,3-thiazinyl, 1,4-thiazinyl, azepinyl, azetidinyl, dioxothiomorpholinyl, imidazolidinyl, morpholinyl, oxanyl, oxazinyl, oxazolidinyl, oxepinyl, oxetanyl, piperazinyl, 2,5-piperazinedionyl,l,4- diasepanyl, piperidinyl, pyranyl, pyrrolidinyl, tetrahydrofuranyl, tetrahydropyranyl, thianyl, thiomorpholinyl or thiopyranyl.In some embodiments for any one or more variable selected from R15, R16, R17, R18, R23, R25, R66, R71, R79, R106 or R113, 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 or spiro[4.5]decanyl.In some embodiments, for any one or more variable selected from R15, R16, R17, R18, R23, R25, R66, R71, R79, R106 or R113, said carbocyclyl or heterocyclyl is independently an optionally substituted group selected from:In some embodiments, for any one or more variable selected from R3, R4, Rll, R12, R15, R16, R17, R18, R47, R62, R63, R64, R65, R67, R68, R69, R70, R74, R75, R76, R77, R78, R105, R108, R109, R110, Rill, R112, R115, R146 (for 3-7 membered carbocyclylenes or heterocyclylenes) or R191, said carbocyclylene or heterocyclylene is independently an optionally substituted group selected from: 1,2-dihydroquinolinediyl, 1,5-naphthyridinediyl, 1,8-naphthyridinediyl, lH-indazolediyl, 1H- isochromenediyl, lH-pyrrolizidinediyl, 1-naphthalenediyl, 2H-benzo[b][l,4]oxazinediyl, 2H- benzo[e][l,2]oxazinediyl, 2h-chromenediyl, 2-naphthalenediyl, 4H-quinolizinediyl, adeninediyl, azaindazolediyl, azaindolediyl, benzimidazolediyl, benzo[b]thiophenediyl, benzo[c][l,2,5]thiadiazolediyl, benzo[c]isothiazolediyl, benzo[c]thiophenediyl, benzo[d]isothiazolediyl, benzo[d]isoxazolediyl, benzo[d]oxazolediyl, benzo[d]thiazolediyl, benzofurandiyl, benzenediyl, cinnolinediyl, cumenediyl, decahydroisoquinolinediyl, decahydroquinolinediyl, guaninediyl, indazolediyl, indenediyl, indolediyl, indolinediyl, indolizinediyl, isobenzofurandiyl, isoindolediyl, isoquinolinediyl, phenylene, phthalazinediyl, pteridinediyl, purinediyl, pyrido[2,3-b]pyrazinediyl, pyrido[4,3-d]pyrimidinediyl, pyrimidinediyl, quinazolinediyl, quinolinediyl, quinoxalinediyl, tetrahydroquinolinediyl, toluenediyl or xylenediyl.In some embodiments for any one or more variable selected from R3, R4, Rll, R12, R15, R16, R17, R18, R47, R62, R63, R64, R65, R67, R68, R69, R70, R74, R75, R76, R77, R78, R105, R108, R109, R110, Rill, R112, R115, R146 (for 3-7 membered carbocyclylenes or heterocyclylenes) or R191, 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, 2,5- piperazinedion-3,6-diyl piperidinediyl, 1,4-diazepandiyl, pyranyl, pyrrolidindiyl, tetrahydrofurandiyl, tetrahydropyrandiyl, thianediyl, thiomorpholinediyl or thiopyrandiyl.In some embodiments for any one or more variable selected from R3, R4, Rll, R12, R15, R16, R17, R18, R47, R62, R63, R64, R65, R67, R68, R69, R70, R74, R75, R76, R77, R78, R105, R108, R109, R110, Rill, R112, R115, R146 (for 3-7 membered carbocyclylenes or heterocyclylenes) or R191, 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 for any one or more variable selected from R3, R4, Rll, R12, R15, R16, R17, R18, R47, 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: 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 for any one or more variable selected from R3, R4, Rll, R12, R15, R16, R17, R18, R47, R62, R63, R64, R65, R67, R68, R69, R70, R74, R75, R76, R77, R78, R105, R108, R109, R110, Rill, R112, R115, R146 or R191, said carbocyclylene is independently an optionaly substituted group selected from:In some embodiments for any one or more variable selected from R3, R4, Rll, R12, R15, R16, R17, R18, R47, R62, R63, R64, R65, R67, R68, R69, R70, R74, R75, R76, R77, R78, R105, R108, R109, R110, Rill, R112, R115, R146 (for 3-7 membered carbocyclylenes or heterocyclylenes) or R191, said carbocyclylene or heterocyclylene is independently an optionally substituted group selected from:In some embodiments for any one or more variable selected from R3, R4, Rll, R12, R15, R16, R17, R18, R47, R62, R63, R64, R65, R67, R68, R69, R70, R74, R75, R76, R77, R78, R105, R108, R109, R110, Rill, R112, R115, R146 (for 3-7 membered carbocyclylenes or heterocyclylenes) or R191, said carbocyclylene or heterocyclylene is independently an optionally substituted group selected from:any aspect / embodiment herein. In some further embodiments, each R33 is independently selected from -H, -CH2(halo), -CH(halo)2, -C(halo)3, -F, -Cl, -Br, -I, -CH3, -OH, -OCH3, -CD3 or -OCD3. In some further embodiments, each R33 is independently selected from -H, -CH2(halo), -CH(halo)2, -C(halo)3, -F, -Cl, - Br, -or -OH.In some embodiments for any one or more variable selected from R3, R4, Rll, R12, R15, R16, R17, R18, R47, R62, R63, R64, R65, R67, R68, R69, R70, R74, R75, R76, R77, R78, R105, R108, R109, R110, Rill, R112, R115, R146 (for 3-7 membered carbocyclylenes or heterocyclylenes) or R191, said carbocyclylene or heterocyclylene is independently an optionally substituted group selected from:In some embodiments for any one or more variable selected from R3, R4, Rll, R12, R15, R16, R17,R18, R47, R62, R63, R64, R65, R67, R68, R69, R70, R74, R75, R76, R77, R78, R105, R108, R109, R110,Rill, R112, R115, R146 or R191, said carbocyclylene or heterocyclylene is independently an optionally substituted group selected from:In some embodiments, -R3-R5-R7-R9-R11-R13-R15, -R3-R5-R7-R9-R11-R13-R17, -R4-R6-R8-R10-R12- R14-R16, and -R4-R6-R8-R10-R12-R14-R18, each independently, have a total number of carbon atoms that is selected from: 16-22, inclusive, 16-20, inclusive, 17, 18 or 19 carbon atoms. In some further embodiments, -R3-R5-R7-R9-R11-R13-R15, -R3-R5-R7-R9-R11-R13-R17, -R4-R6-R8-R10-R12-R14-R16, and -R4-R6-R8-R10-R12-R14-R18, each independently, have a longest linear chain (i.e., excluding side groups) with a total number of carbon atoms that is selected from: 16-22, 16-20, 17, 18 or 19 carbon atoms.In some embodiments, together, -R5-R7-R9- and -R6-R8-R10- are each independently selected from the group consisting of: -O-, -OC(=O)-, -C(=O)O-, -OC(=O)O-, -S-C(=O)-, -C(=O)-S-, -S-S-, -C(=O)N(R164)- , -N(R164)C(=O)-, -N(R164)C(=O)N(R164)-, -O-C(=O)C(R164)2C(=O)O-, -C(=O)O-C(R164)2C(=O)O-, -O- C(=O)C(R164)2-O-C(=O)-, -O-C(R164)2C(=O)O-, -O-C(=O)C(R164)2C-O-, -O-C(R164)2-O-C(=O)-, -C(=O)-O- C(R164)2C-O-, and -O-C(=O)-O-R191-O-C(=O)O-; wherein, R164 and R191 are as defined above or for any aspect or embodiment herein. In some further embodiments, for each occurrence, R164 is independently selected from: -H or optionally substituted C1-C6 alkyl.In some embodiments, one or more of R13, R14, R61, R73 and R107 are each independently selected from:for R13, a bond labelled 2 or 4 is attached to R15 or R62 and a bond labelled 3 is attached to R3, R17 or R67; for R14, a bond labelled 2 or 4 is attached to R16 and a bond labelled 3 is attached to R18; for R61, a bond labelled 2 or 4 is attached to R3, R13, R62 and a bond labelled 3 is attached to R4, 67; for R73, a bond labelled 2 or 4 is attached to R74 and a bond labelled 3 is attached to R76; and for R107, a bond labelled 2 or 4 is attached to R108 and a bond labelled 3 is attached to R110., ,In one aspect, the ionizable cationic lipid has a structural formula represented by (7)(7); wherein, Rll through R18 and R22 through R23 are as defined above.In some embodiments, the ionizable cationic lipid has a structural formula represented by (8):R15R11 — R13R7 — R9 R17R3 — R5R2. ,' NR25R4 — R6R8 — R10 R16R12 — R14R18(8); wherein, each of R2 through R18 and R25 is independently as described above or for any aspect or embodiment herein.In some embodiments, the ionizable cationic lipid has a structural formula represented by (9):(9), wherein, each of R3 through R18 and R25 is independently as described above or for any aspect or embodiment herein.In some embodiments, the ionizable cationic lipid has a structural formula represented by (10):wherein, each of Rll through R18 and R25 is independently as described above or for any aspect or embodiment herein.In some embodiments, the ionizable cationic lipid has a structural formula represented by (11):wherein each occurrence of R2 through R18 is independently as described above or for any aspect or embodiment herein.In another aspect, the ionizable cationic lipid or cationic lipid has is represented by a structural formula selected from:wherein:R142 is selected from: -N(R31)(R32), R58, R119 or R32;R31 and R32 are each independently selected from: -H, -D, -CDs, or an optionally substituted group selected from: linear or branched Ci-io alkyl, CMO hydroxyalkyl, Ci-io alkoxyalkyl, linear or branched Ci- 10 alkenyl, linear or branched CMO alkynyl, 3-14 membered carbocyclyl, 3-14 membered heterocyclyl, 5-10 membered spirocyclyl, 5-10 membered spiroheterocyclyl, 3-7 membered carbocyclyl-Ci.g alkyl-, 3-7 membered heterocyclyl-Ci.g alkyl-, 5-10 membered spirocyclyl-Ci.g alkyl-, 5-10 membered spiroheterocyclyl-Ci.6alkyl-, or, R31 and R32, together with the nitrogen atom to which they are attached, form a 3-14 membered heterocyclyl, 5-10 membered spiroheterocyclyl, 3-7 membered heterocyclyl-Ci-s alkyl- or 5-10 membered spiroheterocyclyl-Ci.g alkyl-;R34 is selected from a bond, -N(R116)-, -O-, -S-, -S(R46);R116 is -H, -D, -CD3, or C1-3 alkyl;R46 is selected from: absent (i.e., two lone electron pairs), =0 or (=0)2;R120 is selected from: a bond, -C(=O)O-, -OC(=O)-, -O-CH2-, -O-, -N(R116)C(=O)-, -C(=O)N(R116)-, - C(=O)O-, -S-S-, -C(=O)- or -C(=O)-CH2-;R47 is selected from: a bond, an optionally substituted 3-14-membered saturated, partially unsaturated or unsaturated carbocyclene or an optionally substituted 3-14-membered saturated, partially unsaturated or unsaturated heterocyclene;R48 is selected from: a bond or an optionally substituted group selected from: CMO alkylene, Ci-w internal alkenylene, C1-10 internal alkynylene, -Co-C3alkyl-3-6-membered cycloalkylene-Co-C3alkyl-, - Co-C3alkyl-4-6-membered cycloalkenylene- Co-C3alkyl-, -Co-C3alkyl-phenylene- Co-C3alkyl-, -Co-C3alkyl-3-6-membered heterocycloalkylene-Co-C3alkyl-, -Co-C3alkyl-4-6-membered heterocycloalkenylene- Co-C3alkyl-, -Co-C3alkyl-5-6-membered heteroarylene- Co-C3alkyl-;R121 is selected from: R134-R135-R136;R134 and R136 are independently selected from: a bond or an optionally substituted group selected from C1-C10 alkylene, C2-C10 alkenylene or C2-C10 alkynylene;R135 is selected from: -C(=O)O-, -OC(=O)-, -OC(=O)O-, -O-, -N(R116)C(=O)-, -C(=O)N(R116)-, - N(R116)C(=O)N(R116)-,-N(R116)C(=O)O-, -OC(=O)N(R116), -S-S- or -C(=O)-;R138 is selected from: R50, R51 or R137. each R50 and R137 are each independently defined above or for any aspect or embodiment herein.R51 is selected from:; wherein,R3-R18, R22, R62 through R71, R73 through R79, R105 through R113, R123 through R128, R132, R133, and any remaining variables are as defined above or for any aspect or embodiment herein;R54 is selected from: -CH2-, -C(=O)-, -C(=S)- or -C(NR59)-, -C(=O)O-, -OC(=O)-, -OC(=O)O-, -O-, - N(R116)C(=O)- or -N(R116)C(=O)-O-; R55 is selected from: a bond, or C1-C4 alkylene;R56 is selected from: a bond, -O-, -S-, -S(=O)-, -S(=O)2- or -NR59-;R57 is a bond, C1-C6 alkylene, -C1-C6 alkylene-O-C1-C6 alkylene-;R58 is selected from:R60 is selected from: is absent (i.e., an unbonded pair of electrons on N), -H, -D, -CD3 or optionally substituted C1-C6 alkyl; wherein, when R60 is -H or -D, -CD3, or optionally substituted C1-C6 alkyl, R60 is bound directly to a nitrogen atom with a +1 charge. The +1 charge may be counterbalanced with an anion from a pharmaceutically acceptable acid; each R139 is independently selected from: -H, -D, -CD3, -OH or C1-C4 alkyl; each R59 is independently selected from: -H, -D, -CD3, C1-C4 alkyl, C1-C4 cycloalkyl, -(CH2)I-4-CI-C4 cycloalkyl or R31; and,R119 is a 4-7-member heterocyclyl, wherein: one ring atom is a nitrogen atom (nitrogen ring atom) and each remaining ring atom is a carbon atom (carbon ring atom); each carbon ring atom adjacent to the nitrogen ring atom is substituted with R31 to form -CH(R31)-; each carbon ring atom that is non- adjacent to the nitrogen ring atom is unsubstituted; said heterocyclyl has a ring with 0 or 1 carbon- carbon double bond, i.e., -CH=CH-; and, when R48 is bound to a carbon ring atom, i.e., when the nitrogen ring atom is not bound to R48, then said nitrogen ring atom is substituted with R31.In some embodiments, the ionizable cationic lipid or cationic lipid is represented by a structural formula of a compound selected from:wherein each of the variables are described above or for any aspect or embodiment herein.The structural formulas of the ionizable cationic lipids or cationic lipids of the present invention may be further defined for any aspect or embodiment as follows:In some embodiments, R22 is selected from: a bond, optionally substituted Ci-5alkylene, optionally substituted -C1-C6 alkylenediyl-O-, -CH2-, -[CHzh-, -[Cl-bh-, isopropylene, butylene, isobutylene, -NH-, - N(R116)-, -O- or -S-. In some embodiments, R22 is -Ci-Cg alkanediyl-O-. In some embodiments, R22 is -C1-C3 alkanediyl-O-. In some embodiments, R22 is selected from: -CH2-O-, -[CHzh-O-, -[CHzh-O-.In some embodiments, R31 and R32 are each independently an optionally substituted group selected from: -CD3, C1-5 alkyl, C1-5 hydroxyalkyl, C1-5 alkoxyalkyl, C1-5 alkenyl, C1-5 alkynyl, or R31 and R32, together with the nitrogen atom to which they are attached, form an optionally substituted aziridinyl, azetidinyl, pyrrolidinyl or piperidinyl group.In some embodiments, R31 and R32 are each independently selected from: -CD3, C1-C4 alkyl, methyl, ethyl, propyl, 2-propyl, cyclopropylmethyl, cyclopropylethyl, cyclopropylpropyl, cyclobutylmethyl, cyclobutylethyl, cyclobutylpropyl, n-butyl, n-pentyl, n-hexyl, hydroxymethyl, 2-hydroxyethyl, 2- hydroxypropyl, 2-hydroxybutyl, 2-hydroxypentyl, 2-hydroxyhexyl, 3-hydroxypropyl, 4-hydroxybutyl, 5-hydroxypentyl, 6-hydroxyhexyl, 2,3-dihydroxypropyl, l,3-dihydroxy-2-propyl, 2,4-dihydroxy-butyl, 2,3,4-trihydroxy-butyl.In some embodiments, R31 and R32 are each independently selected from: 3-7 membered- cycloalkyl, cycloalkenyl, heterocycloalkyl, heterocycloalkenyl.In some embodiments, R31 and R32 are each independently selected from: 3-7 membered- cycloalkyl- C1-C4 alkyl-, 3-7 membered-cycloalkenyl-Ci-C4 alkyl-, 3-7 membered-heterocycloalkyl-Ci-C4 alkyl-, 3-7 membered- heterocycloalkenyl.In some embodiments, R31 and R32 are each independently selected from: -CD3, methyl, ethyl, propyl, hydroxymethyl, hydroxyethyl or hydroxypropyl. In some embodiments, R31 is Ph3C-0-[CH2]o-2CH2- (Ph refers to phenyl). In some embodiments, R31 is hydroxyethyl and R32 is methyl, ethyl or hydroxyethyl. In some embodiments, R31 and R32 are the same. In some embodiments, R31 and R32 ethyl.In some embodiments, R34 is selected from: a bond, -N(R116)- or -O-. In some embodiments, R34 is -O- or -N(R116)-. In some embodiments, R34 is -O- or -NH-.In some embodiments, R47 is an optionally substituted group selected from: 1,2-dihydroquinolindiyl, 1,5-naphthyridindiyl, 1,8-naphthyridindiyl, lH-indazoldiyl, lH-isochromendiyl, lH-pyrrolizidindiyl, 1- naphthalendiyl, 2H-benzo[b][l,4]oxazindiyl, 2H-benzo[e][l,2]oxazindiyl, 2h-chromendiyl, 2- naphthalendiyl, 4H-quinolizindiyl, adenindiyl, azaindazoldiyl, azaindoldiyl, benzimidazoldiyl, benzo[b]thiophendiyl, benzo[c][l,2,5]thiadiazoldiyl, benzo[c]isothiazoldiyl, benzo[c]thiophendiyl, benzo[d]isothiazoldiyl, benzo[d]isoxazoldiyl, benzo[d]oxazoldiyl, benzo[d]thiazoldiyl, benzofurandiyl, benzendiyl, cinnolindiyl, cumendiyl, decahydroisoquinolindiyl, decahydroquinolindiyl, guanindiyl, indazoldiyl, indendiyl, indoldiyl, indolindiyl, indolizindiyl, isobenzofurandiyl, isoindoldiyl, isoquinolindiyl, phenylene, phthalazindiyl, pteridindiyl, purindiyl, pyrido[2,3-b]pyrazindiyl, pyrido[4,3- d]pyrimidindiyl, pyrimidindiyl, quinazolindiyl, quinolindiyl, quinoxalindiyl, tetrahydroquinolindiyl, toluendiyl or xylendiyl.In some embodiments, R47 is an optionally substituted group selected from: furandiyl, imidazoldiyl, isothiazoldiyl, isoxazoldiyl, oxadiazoldiyl, oxazoldiyl, pyrazoldiyl, pyrroldiyl, pyridazindiyl, pyridindiyl, pyrimidindiyl, thiadiazoldiyl, thiendiyl, tetrazoldiyl, thiazoldiyl, triazoldiyl, azepindiyl, azetidindiyl, dioxothiomorpholindiyl, imidazolidindiyl, morpholindiyl, oxandiyl, oxazindiyl, oxazolidindiyl, oxepindiyl, oxetandiyl, piperazindiyl, piperidindiyl, 1,4-diasepandiyl, l,4-piperazin-l,4-diyl-, -1,3- piperazin-l,3-diyl-, l,4-diazepan-l,4-diyl, 2,5-piperazinedion-3,6-diyl, pyranyl, pyrrolidindiyl, tetrahydrofurandiyl, tetrahydropyrandiyl, thiandiyl, thiomorpholindiyl or thiopyrandiyl.In some embodiments, R47 is an optionally substituted group selected from: cyclopropylene (cyclopropandiyl), cyclobutylene (cyclobutandiyl), cyclopentylene (cylcopentandiyl), cyclohexylene (cyclohexendiyl), cycloheptylene (cycloheptandiyl), cyclopropendiyl, cyclobutenylendiyl, cyclopentenylendiyl, cyclohexendiyl, cycloheptendiyl, cyclobutadiendiyl, cyclopentadiendiyl, cyclohexadiendiyl or cycloheptadiendiyl or cycloheptatriendiyl.In some embodiments, R47 is an optionally substituted group selected from: adamantandiyl, azabicyclo[3.1.0]hexandiyl, 3-azabicyclo[3.1.1]heptandiyl, bicyclo[2.1.1]hexyl, bicyclo[2.2.1]heptyl, bicyclo[2.2.1]heptenyl, 3-oxa-8-azabicyclo[3.2.1]octandiyl, 6-oxa-3-azabicyclo[3.1.1]heptandiyl, 8- oxa-3-azabicyclo[3.2.1]octandiyl, 3-oxa-6-azabicyclo[3.1.1]heptandiyl, 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]hexandiyl, 2- azaspiro[3.3]heptandiyl, 2-oxa-6-azaspiro[3.3]heptandiyl, spiro[2.2]pentanyl, spiro[3.3]heptanyl, spiro[2.3]hexandiyl, spiro[2.5]octandiyl or spiro[4.5]decandiyl.In some embodiments, R47 is an optionally substituted group selected from:, wherein stereochemical relationship of the substituent groups on each ring can be cis or trans.In some embodiments, R47 is an optionally substituted group selected from:, ,In some embodiments, R47 is an optionally substituted group selected from:In some embodiments, R47 is an optionally substituted phenylene.In some embodiments, R47 is an optionally substituted group selected from:In some embodiments, R47 is independently substituted with one or more R144, where R144 is selected from: -D, -CH2CH3,-CH2(halo), -CH(halo)2, -C(halo)3, -F, -CF3, -Cl, -Br, -I, -CH3, -OCH3, -CD3, - OCD3, -CH2OCH3, -CH2R25, -CH(R25)2, -C(R25)3, -OH, -OR25, -CN, -NO2,-NH2, -NHR25, -N(R25)2, -SH, - SR25, -S(=O)H, -S(=O)R25, -S(=O)2H, -S(=O)2R25, S(=O)2CH3, -PH2, -PHR25, -P(R25)2, -P(OH)2, -POHR25,-P(R25)2, -P(OR25)2, -P(O)H2, -P(O)HR25, -P(O)(R25)2, -P(O)(OH)2, -P(O)(OR25)OH, -P(O)(OR25)2, -SiH3,-SiH2R25, -SiH(R25)2, or -Si(R25)3. In some further embodiments, each R144 is independently selected from: -F, -CF3, -Cl, -Br, -I, -OH, -S(=O)2CH3, -S(=O)2R25, -NHCH3or -NHR25. In some further embodiments, each R144 is selected from: -F, -Cl, -Br, -OCH3or -OH. In some further embodiments, the R47 carbocyclene or heterocyclene is independently substituted with 1, 2, 3, 4 or 5 R144 groups. In some further embodiments, each R144 is independently selected from: -D, -OH, -OCD3, -OCH3,-F, Cl or Br. In some further embodiments, at least one R144 is -F; at least one R144 is -Cl; at least one R144 is -Br; at least two R144 are -F; at least two R144 are -Cl; at least two R144 are -Br; at least one R144 is -OH; at least two R144 are -OH; at least one R144 is -F -Cl or -Br and at least one R144 is -OH or - OCH3; at least two R144 is independently selected from -F, -Cl or -Br and at least one R144 is -OH or - OCH3; at least two R144 is -F and at least one R144 is -OH; at least two R144 are -Cl and at least one R144 is -OH or -OCH3; at least two R144 are -Br and at least one R144 is -OH; at least two R144 is independently selected from -F, -Cl or -Br; or at least 3 R144 are independently selected from -F, -Cl or -Br.In some embodiments, R48 is an optionally substituted group selected from: Ci-Cg alkylene, C2-C8 internal alkenylene, C2-C8 internal alkynylene, -Co-C3alkyl-3-6-membered cycloalkylene-Co-C3alkyl-, - C0-C3alkyl-4-6-membered cycloalkenylene- C0-C3alkyl-.In some embodiments, R48 is an optionally substituted group selected from: methylene, ethylene, propylene, C3-Cg branched alkylene, -Co-C3alkylene-3-6-membered cycloalkylene-Co-C3alkylene- or Co-C3alkylene-4-6-membered cycloalkenylene- Co-C3alkylene-.In some embodiments, R48 is -Co-C3alkylene-3-6-membered cycloalkylene-Co-C3alkylene-, wherein said 3-6-membered cycloalkylene is selected from cyclopropylene, cyclobutylene, cyclopentylene or cyclohexylene.In some embodiments, R48 is an optionally substituted group selected from: methylene, ethylene, propylene, isopropylene, butylene or isobutylene.In some embodiments, R57 is selected from: methylene, ethylene or propylene. In some embodiments, R57 is -[CH2]I.5-O-[CH2]I.5-. In some embodiments, R57 is -[CH2]I.3-O-[CH2]I.3-. In some embodiments, R57 is selected from: -[CH2]2-O-[CH2]3- or -[CH2]3-O-[CH2]2-.In some embodiments, R120 is selected from: a bond, -C(=O)O-, -OC(=O)-, -C(=O)C-, -O-. - C(=O)N(R116)-, -N(R116)C(=O)-.In some embodiments, R54 is selected from: -C(=O)-, -OC(=O)- or -NHC(=O)-.In some embodiments, R119 is a heterocycloalkyl. In some embodiments, R119 is heterocyclyl selected from azetidinyl, pyrrolidinyl, piperindinyl or azepanyl.In some embodiments, R119 is selected from:In some embodiments, R59 is -H, -D, -CD3, or C1-C4 alkyl. In some further embodiments, R59 is selected from: -H, -D, -CD3, methyl, ethyl, propyl, isopropyl, or cyclopropylmethyl. In some further embodiments, R59 is selected from: -H, -D, -CD3, -OH, methyl or -[CHjh-j-OH.In some embodiments, R31 and / or R32 are each independently substituted with one or more substituents each independently selected from: R15 or R24; wherein R15 and R24, for each occurrence, is independently as described above or for any aspect or embodiment herein.In some embodiments, R31 and / or R32 are each independently substituted with one or more substituents each independently selected from: a C3-Ci4-membered partially unsaturated or unsaturated carbocycle or a 3-14-membered partially unsaturated or unsaturated heterocycle containing one or more of the heteroatoms N, O and / or S, wherein the carbocycle or heterocycle optionally substituted with one or more substituents selected from: R15, or R24; wherein R15 and R24, for each occurrence, is independently as described above or for any aspect or embodiment herein.In some embodiments, R134 and R136 are independently selected from: a bond or an optionally substituted group selected from: C1-C6 alkylene, C2-C6 alkenylene or C2-C6 alkynylene. In some embodiments, R134 and R136 are independently selected from: a bond or an optionally substituted group selected from: C1-C3 alkylene, C2-C3alkenylene or C2-C3alkynylene. In some embodiments, R134 and R136 are independently selected from: methylene, ethylene or propylene. In some embodiments, R134 and R136 are independently selected from: methylene or ethylene.In some embodiments, R134 and R136 are independently selected from: a bond or an optionally substituted group selected from C1-C6 alkylene, C2-C6 alkenylene or C2-C6 alkynylene. In some embodiments, R134 and R136 are independently selected from: a bond or an optionally substituted group selected from C1-C6 alkylene. In some embodiments, R134 and R136 are independently Ci-C3alkylene.In some embodiments, R135 is selected from: -C(=O)O-, -OC(=O)-, -OC(=O)O-, -O-, - or -C(=O)-. In some embodiments, R135 is selected from: -C(=O)O-, -OC(=O)- or -OC(=O)O-.In some embodiments, R121 is selected from: -C0-C3 alkylene-C(=O)O- Co-C3alkylene-, -Co-C3alkylene- OC(=0)-Co-C3alkylene-, -Co-C3alkylene-OC(=0)0-Co-C3alkylene-, -Co-C3alkylene-0-Co-C3alkylene-, -Co- C3alkylene-N(R116)C(=O)-C0-C3alkylene-, -C0-C3alkylene-C(=O)N(R116)-C0-C3alkylene-, -C0-C3alkylene-N(R116)C(=O)N(R116)-C0-C3alkylene-, -C0-C3alkylene-N(R116)C(=O)O-C0-C3alkylene-, -C0-C3alkylene-OC(=0)N(R116)-Co-C3alkylene-, -Co-C3alkylene-S-S-Co-C3alkylene- or-Co-C3alkylene-C(=O)- Co-C3alkylene-. In some embodiments, R121 is selected from: -C1-C3 alkylene-C(=O)O- C1-C3 alkylene-, -C1-C3 alkylene-OC(=O)-C1-C3 alkylene-, -C1-C3 alkylene-OC(=O)O-C1-C3 alkylene-, -C1-C3 alkylene-O-Ci- C3alkylene-, -C1-C3 alkylene-N(R116)C(=O)-C1-C3 alkylene-, -C1-C3 alkylene-C(=O)N(R116)-Ci-C3alkylene-, -C1-C3 alkylene-N(R116)C(=O)N(R116)-C1-C3 alkylene-, -C1-C3 alkylene-N(R116)C(=O)O-Ci-C3alkylene-, -C1-C3 alkylene-OC(=O)N(R116)-C1-C3 alkylene-, -C1-C3 alkylene-S-S-C1-C3 alkylene- or-Ci-C3alkylene-C(=O)-C1-C3 alkylene-. In some embodiments, R121 is selected from: -C1-C3 alkylene-C(=O)O- C1-C3 alkylene-, -C1-C3 alkylene-OC(=O)-C1-C3 alkylene-, -C1-C3 alkylene-OC(=O)O-C1-C3 alkylene- or -Ci- C3alkylene-O-C1-C3 alkylene-.In some embodiments, R121 is selected from: -CH2OC(=O)OCH2-, -[CH2]3OC(=O)OCH2-,In some embodiments, the ionizable cationic lipid or cationic lipid represented by any one of structural formulas (12)-(20), (21) (23), (24), (27), (29), (30) or (31) wherein (as present):R31 and R32 are ethyl, R31 is hydroxyethyl and R32 is -CD3or methyl, or R31 is Ph3C-O-CH2-CH2- and R32 is -CD3or methyl;R48 is propylene or butylene;R34 is -O- or -NH-;R47 is benzene-l,3-diyl or benzene-l,4-diyl;R22 is methoxy;R120 is -OC(=O)-;R121 is -CH2OC(=O)OCH2-; and each remaining variable is as defined above or for any aspect or embodiment hereinIn some embodiments, the ionizable cationic lipid or cationic lipid represented by any one of structural formulas (15), (19), (24), (30), (31) wherein:R32 is Ci-io hydroxyalkyl;R121 is -Ci-Cio alkylene-OC(=O)O- Ci-Cio alkylene-; and each remaining variable is as defined above or for any aspect or embodiment herein.In some embodiments, ionizable cationic lipid or cationic lipid is represented by structural formula (20)(20); wherein,R31 and R32 are each independently selected from: -CDs, Ci to C3 alkyl, cyclopropylmethyl, 2- hydroxyethyl, 2-hydroxypropyl, or 3-hydroxypropyl;R48 is selected from: a bond, or Ci to C6alkylene;R34 is selected from: a bond, -O-, -NH-, or -S-;R22 is selected from: a bond, substituted Ci to C3alkylene, -CH2-O-, -[Cl-bh-O-, -[CH2]3-O-, -CH2-S-, - [CH2]2-S-, or -[CH2]3-S-.In some embodiments, ionizable cationic lipid or cationic lipid is represented by structural formula (24)R31 and R32 are each independently selected from: -CD3, Cito C3alkyl, hydroxymethyl, 2- hydroxyethyl, 3-hydroxypropyl, or Ph3C-0-[CH2]o-2CH2- (Ph refers to phenyl);R48 is selected from Ci to C7alkylene;R22 is selected from Ci to C3alkylene;R120 is selected from: -O-, -O-C(=O)-, -C(=O)-O-, -O-C(=O)-O-, -C(=O)-NH-, -HN-C(=O)-, or -S-C(=O)- NH-, -S(=O)2-NH-, -O-S(=O)2-NH-;R121 is selected from: -CHC0-C3] alkylene, -0-C(=0)-[Co-C3] alkylene, -C(=0)-0-[Co-C3] alkylene, -O- C(=O)-O-[C0-C3] alkylene, -C(=O)-NH-[C0-C3] alkylene, -NH-C(=O)- [C0-C3] alkylene, or -S-C(=O)-NH-[C0- C3] alkylene-.In further embodiments of structural formula (24),R31 is selected from -CD3or Ci to C3alkyl,R32 is selected from 2-hydroxyethyl, 2-hydroxypropyl, 3-hydroxypropyl, cyclopropylmethyl or PhsC- 0-[CH2]O-2CH2- (Ph refers to phenyl),R48 is selected from C2to C5alkylene,R120 is selected from: -O-, -O-C(=O)-, or -C(=O)-O-;R121 is selected from: -O-C(=O)-O-(CH2)I.2-, -C(=O)-NH-(CH2)I.2-, -NH-C(=O)-(CH2)I.2- or-S-C(=O)-N-(CH2)I.2-; and,R22 is selected from -CD3 or Ci to C3 alkylene.In some embodiments, ionizable cationic lipid or cationic lipid is represented by structural formula (25):wherein,R58 is selected fromR57 is selected from a bond, -CD3or Ci to C5alkylene;R54 is selected from -O-C(=O)-, -C(=O)-O-, -NH-C(=O)-, -C(=O)-NH-, -S-C(=O)-, -S(=O)2-NH-, -O-S(=O)2- NH-;R59 and R139 are each independently selected from: -H, -D, -CD3, C1-C3 alkyl or C1-C3 alkylene-OH; andR60 is -H or -D.In some embodiments, ionizable cationic lipid or cationic lipid is represented by structural formula (26):(exo conformation), (endo conformation),R54 is selected from -O-C(=O)-, -C(=O)-O-, -NH-C(=O)-, -C(=O)-NH-, -S-C(=O)-;R59 and R139 are each independently selected from: -H, -D, -CD3, C1-C3 alkyl or C1-C3 alkylene-OH; andR60 is -H or -D. In some embodiments, ionizable cationic lipid or cationic lipid is represented by structural formula (29)R31 and R32 are each independently selected from: -CD3, Ci to C4alkyl, cyclopropylmethyl, 2- hydroxyethyl, 2-hydroxypropyl, 3-hydroxypropyl, or 4-hydroxybutyl;R48 is selected from a bond, or Ci to C6alkylene;R120, R47, and R22 are each a bond; and,R121 is selected from: -C0-C3 alkylene-0-C(=0)-0-[Co-C3] alkylene-, -C0-C3 alkylene-C(=0)-0-[Co-C3] alkylene -, -C0-C3 alkylene-0-C(=0)-[Co-C3] alkylene-, -Co-C3 alkylene-C(=0)-NH-[Co-C3] alkylene- or -Co- C3alkylene-NH-C(=0)-[Co-C3] alkylene-.In some embodiments, ionizable cationic lipid or cationic lipid is represented by structural formula (34):wherein,independently selected from: -H, -D, -F, -CF3, -Cl, -Br, -I, -OH, -OCH3,-OCD3-, CH2S(=O)-; or -CH2S(=O)2-R22 is selected from a bond (absent), Ci to C5alkylene, -Co-C2alkylene-C(=O)- [C0-C4] alkylene-, hydroxymethyl, 1-hydroxyethyl, or 1-hydroxypropyl;R21 is selected from: -O-, -O-C(=O)-, -C(=O)-O-, -O-C(=O)-O-, -C(=O)-NH-, -NH-C(=O)-, or -S-C(=O)-NH-, or -NH-C(=O)-S-; and,R62 is selected from Ci to C5alkylene.In some further embodiments of structural formula (33):R23 is selected from: fR22 is selected from: Ci to C3alkylene, hydroxymethyl, 1-hydroxyethyl, or 1-hydroxypropyl;R21 is selected from: -C(=O)-N-, -N-C(=O)-, or -S-C(=O)-N-, -N-C(=O)-S-; and,R62 is selected from Ci to Cg alkylene.In some further embodiments of structural formula (33):R21 is selected from: -C(=O)-NH-, -NH-C(=O)-, or -S-C(=O)-N-, -N-C(=O)-S-; and,R62 is selected from: a bond or Ci to Cg alkylene.In some embodiments, ionizable cationic lipid or cationic lipid is represented by structural formula (35):R23 is selected from:R22 is selected from: a bond, C1-C3 alkylene, hydroxymethyl, 1-hydroxyethyl, or 1-hydroxypropyl;R21 is selected from: a bond, -C(=O)-, -NH-, -C(=O)-NH-, or -NH-C(=O)-; R62 is selected from: a bond, Ci to C5alkylene;R120 is selected from : a bond, -O-, -O-C(=O)-, -C(=O)-O-, -O-C(=O)-O-, -C(=O)-N-, -N-C(=O)-, -S-C(=O)- N-, or -N-C(=O)-S-; andR67 is selected from Ci to Cg alkylene.In some embodiments, ionizable cationic lipid or cationic lipid is represented by structural formula (36)R25 is selected from: branched or linear Ci to Cg alkyl, -CD3, cyclopropyl, cyclobutyl, or branched or linear hydroxy-Ci-C8alkyl-; andR21 is a bond.In some embodiments, each R50, R51, R138 and R137 (where applicable) is represented by a structural formula independently selected from:5 In some embodiments, each R50, R51, R138 and R137 (where applicable) is represented by a structural formula independently selected from:wherein R61 is defined above or for any aspect or embodiment herein. In some embodiments, R61 is selected from:In some embodiments, each R50, R51, R138 and R137 (where applicable) is represented by a structural formula independently selected from:In another aspect, the ionizable cationic lipid or cationic lipid has a structural or chemical formula of a compound of Table 1., wherein R138 is selected from R50, R51 or R137, wherein each R50, R51 and R137 are defined above or for any aspect or embodiment herein. In some embodiments, R138 is R50. In some embodiments R138 is R51. In some embodiments R148 is R137.Table 1.wherein, each R50, R51, R59, R137, R138 and any remaining variables are as described above or any other embodiment herein.In some embodiments, the ionizable cationic lipid has a structural formula selected from a compound in Table 2.Table 2In another aspect, the invention provides for a cationic lipid with a structural formula of a compound in Table 3. Table 3In some embodiments the 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 intracellular 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 phosphate-buffered saline (PBS), at a concentration of 0.4 mM total lipids, 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 values. 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, after vortex mixing, the 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 non-linear least-squares fit analysis (titration curve) 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. Nonlimiting examples of cholesterol derivatives include 5-a-cholestanol (5a-Cholestan-3p-ol), 5-p- 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 someembodiments, cholesterol derivative is cholesteryl hemisuccinate. In some embodiments, the sterol is a sea cucumber sulphated sterol, e.g., cholest-5-en-3P-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, distearoyl-phosphatidylcholine (DSPC), dioleoyl-phosphatidylcholine (DOPC), dipalmitoyl-phosphatidylcholine (DPPC), dioleoyl-phosphatidylglycerol (DOPG), dipalmitoylphosphatidylglycerol (DPPG), dioleoyl-phosphatidylethanolamine (DOPE), palmitoyl-oleoyl- phosphatidylcholine (POPC), palmitoyl-oleoyl-phosphatidylethanolamine (POPE), dioleoylphosphatidylethanolamine 4-(N-maleimidomethyl)-cyclohexane-l-carboxylate (DOPE-mal), dipalmitoyl phosphatidyl ethanolamine (DPPE), dimyristoyl-phosphoethanolamine (DMPE), distearoyl-phosphatidyl-ethanolamine (DSPE), monomethyl-phosphatidylethanolamine (such as 16- O-monomethyl PE), dimethyl-phosphatidylethanolamine (such as 16-O-dimethyl PE), 18-1-trans PE, 1- stearoyl-2-oleoyl-phosphatidyethanolamine (SOPE), hydrogenated soy phosphatidylcholine (HSPC), egg phosphatidylcholine (EPC), dioleoyl-phosphatidylserine (DOPS), sphingomyelin (SM), 1,2- diundecanoyl-sn-glycero- phosphocholine (DUPC), dimyristoyl phosphatidylcholine (DMPC), dimyristoyl phosphatidylglycerol (DMPG), distearoyl-phosphatidylglycerol (DSPG), dierucoyl- phosphatidylcholine (DEPC), palmitoyl-oleoyl-phosphatidylglycerol (POPG), dielaidoylphosphatidylethanolamine (DEPE), 1,2- dilauroyl-sn-glycero-3-phosphoethanolamine (D LPE); 1,2- diphytanoyl-sn-gl ycero-3- phosphoethanolamine (DPHyPE); l,2-di-O-octadecenyl-sn-glycero-3-phosphocholine (18:0 Diether PC), l-oleoyl-2-cholesterylhemisuccinoyl-sn-glycero-3-phosphocholine (OChemsPC), l-hexadecyl-sn-glycero-3-phosphocholine (C16 Lyso PC), l,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), 1,2- distearoyl-sn-glycero-3-phosphoethanolamine, 1.2-dilinoleoyl-sn-glycero-3-phosphoethanolamine, l,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, egg sphingomyelin (ESM), dihydrosphingomyelin, cephalin, cardiolipin, phosphatidicacid, cerebrosides, dicetylphosphate, lysophosphatidylcholine, dilinoleoyl- 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 Ci0-C24carbon chains, e.g., lauroyl, myristoyl, palmitoyl, stearoyl, or oleoyl. 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 WO2017 / 099823 and US Patent Application Publication 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 non-cationic 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, polysarcosine), 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 lipids include, 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) 1,2-dimyristoyl-rac-glycero-3-methoxypolyethylene glycol-2000 (PEG2000-DMG), R-3-[(co-methoxy poly(ethylene glycol)2000)carbamoyl)]-l,2-dimyristyloxy-propyl-3-amine (PEG-c-DOMG), 1,2-Dimyristoyl-sn- Glycero-3-Phosphoethanolamine (DMPE) conjugated Polyethylene Glycol (PEG-DMPE), 1,2-Dilauroyl- sn-Glycero-3-Phosphoethanolamine (DLPE) conjugated Polyethylene Glycol (PEG-DLPE), dipalmitoyl phosphatidylcholine (DPPC) conjugated Polyethylene Glycol (PEG-DPPC), 1, 2-Distearoyl-sn-glycero-3- phosphoethanolamine-Poly(ethylene glycol) (PEG-DSPE), 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 methoxy on the PEG terminus opposite to the hydrophobic lipid end, as inPEG with mono-tail can be an amide, as inThis lipid may be used in LNP formulations, usually with a PEG2k segment, as defined below, as a sole surface stabilizer or in mixtures with other surface lipids. The number of ethyleneoxy (-CH2CH2O-) repeating units in PEG polymers is usually referred to as n, often expressed as a range due to polydispersity of the preparation. Thus, the values of n for a 2000 molecular weight (2k) PEG is between about 40-50 ethyleneoxy units. The value of n is smaller for lower molecular weight PEG and larger when the PEG size is increased relative to PEG2000, also referred to as PEG2k.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 entirety.In one embodiment, the PEG-DAA PEGylated lipid can be, for example, PEG-dilauryloxypropyl, PEG- dimyristyloxypropyl, PEG-dipalmityloxypropyl, or PEG-distearyloxypropyl. The PEG-lipid can be one or more of PEG-DMG, PEG-dilaurylglycerol, PEG-dipalmitoylglycerol, PEG-distearylglycerol, PEG- dilaurylglycamide, PEG-dimyristylglycamide, PEG-dipalmitoylglycamide, PEG- distearylglycamide, 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), and 1,2-dimyristoyl-snglycero- 3-phosphoethanolamine-N- [methoxy(polyethylene glycol)-2000]. In one embodiment, the PEG-lipid can be selected from thegroup consisting of PEG-DMG, and l,2-dimyristoyl-sn-glycero-3- phosphoethanolamine-N- [methoxy(polyethylene glycol)-2000].In some embodiments, the PEGylated lipid is selected from N-(Carbonyl-methoxypolyethyleneglycoln)- l,2-dimyristoyl-sn-glycero-3-phosphoethanolamine (PEGn-DMPE, where PEG average molecular weight is 350, 500, 750, 1000 or 2000); N-(Carbonyl-methoxypolyethyleneglycoln)-l,2-distearoyl-sn- glycero-3-phosphoethanolamine (DSPE-PEGn, where PEG average molecular weight is 350, 500, 750, 1000, 2000, or 5000); DSPE-polyglycerol-cyclohexyl-carboxylic acid, DSPE-polyglycelin-2- methylglutaric acid; l,2-Distearoyl-sn-Glycero-3-Phosphoethanolamine (DSPE) conjugated Polyethylene Glycol (HO-PEG-DSPE); polyethylene glycol-dimyristolglycerol (PEG-DMG); polyethylene glycol-distearoyl glycerol (PEG-DSG); or N-octanoyl-sphingosine-l-{succinyl[methoxy(polyethylene glycol)2000 (Cg PEG2000 Ceramide). In some examples of PEGn-DMPE, where PEG average molecular weight is 350, 500, 750, 1000 or 2000, the PEG-lipid is N-(Carbonyl-methoxypolyethyleneglycol 2000)- l,2-dimyristoyl-sn-glycero-3-phosphoethanolamine (PEG2000-DMPE). In some examples of PEGn- DSPE, where 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 (PEG2000-DSPE). In some embodiments, the PEGylated lipid is HO-PEG-DSPE. In some embodiments, the PEGylated lipid is azide-PEG-DSPE. 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, PEG-DSPE conjugated with a GalNAc ligand, PEG-DSG 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 cationic -polymer lipid (CPL) conjugates 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.In some embodiments, the PEGylated lipid isIn another aspect, the invention provides for a PEG-lipid with a structural formula of a compound in Table 4.Table 4. PEG2k lipid conjugates.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 2.0% 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 one embodiment, the lipid particles (e.g., lipid nanoparticles) may be 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 polysarcosine), and mixtures thereof. Additional examples of POZ-lipid conjugates are described in PCT Publication No. WO 2010 / 006282. PEG 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., non-ester 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 LipidsThe lipid particles of the present invention may further comprise one or more additional lipid(s), including triglycerides and fatty acids. 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.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 single-stranded DNA, singlestranded 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), enhancer RNA (eRNA), aptamers, 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, transposable elements, including retrotransposons, e.g., class I (e.g., Long Terminal Repeat (LTR) Retrotransposons and Non-LTR Retrotransposons, e.g., LINEs (Long Interspersed Nuclear Elements and SINEs (Short Interspersed Nuclear Elements)), Long Serine Recombinase (LSR), and class II (e.g., simple transposons (insertion sequences), composite transposons, and replicative transposons) retrotransposons and non-autonomous elements; and any combination of the above. Examples of class II include Sleeping Beauty (SB) Transposon System (Tcl / mariner-type DNA transposon), PiggyBac transposon and Tol2 transposon 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 preparation. Alternatively, LNP preparations containing different TNAs may be combined into a single formulation.The LNPs of the invention are useful in methods of gene therapy and may comprise, mRNA encoding a therapeutic protein of interest or a protein / enzyme enabling the method. By way of example, the mRNA may encode: 1. Genome editing enzymes, including: a) endonucleases (e.g., ZFN (Zinc Finger Nucleases), TALEN (Transcription Activator-Like Effector Nucleases), Cas9 (CRISPR-Cas9), Casl2a (Cpfl), and CasX, CasY, Casd) (theta)); b) Base Editors (e.g., cytidine deaminase (e.g., APOBEC1), adenosine deaminase (e.g., TadA), and Cas9 nickase fused with deaminase); c) Prime Editors (e.g., Cas9 nickase + reverse transcriptase (RT)); 2. Gene addition and vector integration enzymes, including: a) Integrases (e.g., HIV-1 integrase, Cp(theta)C31 integrase, Bxbl integrase, TP901-1 integrase and A118 integrase; b) Transposases (e.g., PiggyBac Transposase, Sleeping Beauty Transposase (SB100X) and Tol2 Transposase); 3. RNA-based enzymes in gene therapy, including: a) RNA-targeting CRISPR Enzymes Casl3a, Casl3b, Casl3d; b) Reverse transcriptase (RT) (e.g., Moloney Murine Leukemia Virus RT (M-MLV RT) and HIV-1 RT); and c) RNA deaminases (e.g., ADAR (Adenosine Deaminases Acting on RNA), REPAIR system and LEAPER); 4. DNA repair enzymes (e.g., DNA Ligase IV, Rad51, BRCA1, BRCA2 and POLQ); and 5. Nickases (e.g., Cas9n (Cas9 nickase). Additional payload nucleic acids used in the methods include: sgRNA / crRNA (CRISPR RNA) / tracrRNA (trans-activating crRNA)(CRISPR (Cas9, Casl2)), pegRNA / RNA donor templates (Prime editing), gRNA for Casl3 (RNA guide)(CRISPR-Casl3), ssODN / dsDNA (CRISPR, ZFN, TALEN), ASO (antisense oligonucleotides) / miRNA mimics (SMA, cancer, rare diseases), AAV donor template / plasmid DNA (Viral vector systems), Transposon DNA (with ITRs) (Sleeping Beauty, PiggyBac), circRNA / saRNA (LNP-based delivery), scaffold RNA (SAM, scRNA)(CRISPRa / i), and attP / attB DNA sites (site-specific integration (e.g., (PC31 integrase).According to some embodiments, the LNP has a total lipid to TNA mass 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 untranslated region (UTR) 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 more alternative 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-pseudo uridine or 1-ethyl-pseudo uridine), 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).uAn 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- uracil (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-pseudo uridine, 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-pseudo uridine, 5-taurinomethyl-2-thio-uracil (rm5s2U), l-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-pseudo uridine, 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), l-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- 1-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. Exemplary nucleobases 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, l-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-thiouracil, 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 anupstream 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 via antigen presentation by professional antigen presenting cells (APCs) or other presenting cells including somatic cells and followed by a humoral response mediated by B-cells. 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 (Evonik). In some cases, the extrusion step can be omitted. Ethanol removal and simultaneous buffer exchange 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.A general small-scale formulation can be prepared as follows: An mRNA solution and an anhydrous ethanol lipid solution are prepared for mixing. mRNA / acidic buffer mRNA stream is prepared in acidic buffer at a concentration between 0.05 to 0.25 mg / mL in either sodium acetate, sodium citrate or other appropriate acid with buffer at a molar concentration ranging from about 10 to about 100 mM at a pH between 3 to 6. For certain formulations, sodium chloride (NaCI) can be added to the acidic buffer up to 150 mM.Lipids / ethanolLipids are a mixture of several components dissolved in anhydrous ethanol. The lipid mixture typically comprises four components: ionizable lipid(s), phospholipid(s) (e.g. DSPC and DOPE), sterol(s) (e.g.cholesterol), and polymer conjugated lipid(s) (e.g. PEG-lipid such as DMG-PEG2k). The molar concentration range of each component is shown below.• Ionizable lipid: 30 to 60%• Sterol: 30 to 50%• Phospholipid: 5 to 25%• PEG-lipid: 1 to 3%The concentration of the lipid mixture is a function of the N:P ratio. The N:P ratio is the molar ratio of ionizable amine of ionizable lipids (N) to phosphate group of mRNA (P). The N:P ratio generally ranges from 3 to 6. The lipid mixture concentration in ethanol generally ranges from 3 to 18 mg / mL. The mixing volume ratio of lipid / ethanol to mRNA / acidic buffer is typically 1 to 3.Mixing conditions mRNA stream and lipid stream can be mixed through different types of mixers such as microTee mixer, microfluidics, vortex mixer. In some cases, pipette mixing can also be adapted for a very small-scale formulation. For instance, lipids / ethanol can be added into mRNA / acidic buffers using a pipette or liquid handler. The mixing volume of lipid / ethanol to mRNA / acidic buffer is typically 1:3.Buffer exchangeAfter mixing is complete, buffer exchange will be performed to rapidly reduce the level of ethanol percentage and increase pH. Buffer exchange can be done through performing dialysis, using a desalting column combined with centrifuge, or dilution with a storage buffer.Concentration and sterile filtrationConcentration is needed if a higher concentration than the output is required. For a small-scale formulation, concentration is typically carried out using a centrifugal filter combined with centrifuge. Finally, sterile filtration is conducted through passing the LNP solution through a 0.22 um sterile filter.General formulation composition examplesAccording 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, non-viscous and opaque nanosuspension. 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 filtration 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 delivery 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 ionizable 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, double-stranded, triplex and quadriplex 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, anticacking and humectants, deflocculating agents and / or bulking crosslinked polyvinyl pyrrolidone (crospovidone), cellulose, methylcellulose, carboxymethyl cellulose, cross-linked sodium carboxymethyl cellulose (croscarmellose), magnesium aluminum silicate (VEEGUM®), 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), sugar cyclic analogs (cyclodextrins, alpha, beta, gamma, delta), 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 and chelators 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, maleic acid, 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, sodiumpropionate, 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 acid salts), 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, saccharides, dextrans, cyclodextrins, lipoplexes, microvesicles, exosomes, 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 or spray device. 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 μg to about 50 mg, about 0.02 μg to about 0.2 μg, or about 0.2 μg to about 2.0 μg, about 1 μg to about 25 μg, about 25 μg to about 50 μg, about 50 μg to about 100 μg, about 100 μg to about 200 μg, about 200 μg to about 300 μg, about 300 μg to about 400 μg, about 400 μg to about 500 μg, about 500 μg to about 750 μg, about 750 μg 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, about 50 mg to about 100 mg.Methods of UsingThe pharmaceutical compositions comprising a lipid nanoparticle (LNP) 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 a lipid nanoparticle (LNP) 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, disorder or condition 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 an LNP and a TNA or combination of TNAs.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 disease, disorder or condition 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 an 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 production of a functional protein or increased expression of a protein, such as OTC enzyme, CPS1 enzyme, GSDla 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, granulocytemacrophage 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, bispecific, 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, transmucosal, intranasal, inhalation (e.g., via an aerosol, soft-mist or dry powder), buccal (e.g., sublingual), vaginal, intrathecal, intraocular, transdermal, intraendothelial, in utero (or in ovo), parenteral (e.g., intravenous, subcutaneous, intradermal, intracranial, intramuscular [including administration to skeletal, diaphragm and / or cardiac muscle], intrapleural, intracerebral, and intraarticular), 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 organ injection (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, intraarterial 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 directintramuscular 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, intraocular, 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, once or multiple times.ExamplesSynthesis of ionizable and cationic lipidsSynthesis 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.Intermediate Lipids A, B, C, E, F, G, I, J, Core 1, Core 2, Core 3, Core 4, Core 5, Core 6, Core 11Synthesis of 6-((6-((2-hexylnonanoyl)oxy)hexyl)amino)hexyl 2-hexyldecanoate (Lipid A):Synthetic Scheme:Step 1: Synthesis of 6-bromohexyl acetate (A-2)To a stirred solution of 6-bromohexan-l-ol (A-l) (10.0 g, 55.23 mmol) in toluene (150 mL) at 0 °C, was added Na2CO3(7.0 g, 66.27 mmol), DMAP (1.35 g, 5.52 mmol) followed by acetic anhydride (6.0 mL, 63.52 mmol) and the reaction mixture was stirred at 0° C for 1 h (monitored byTLC). Then the reaction mixture was quenched with IN HCI (250 mL) and extracted with EtOAc (2 x 500 mL). The combined organic layer was washed with water (500 mL), dried over anhydrous Na2SO4, filtered and the filtrate was concentrated under vacuum to afford 6-bromohexyl acetate (A-2) (11.5 g, 51.54 mmol, 93 % yield) as a colorless liquid.TH NMR (400 MHz, DMSO-d6): 6 ppm 3.97 (t, J = 3.2 Hz, 2H), 3.53-3.50 (m, 2H), 1.99 (s, 3H), 1.81-1.76 (m, 2H), 1.58-1.54 (m, 2 H), 1.45-1.28 (m, 4H).Step 2: Synthesis of (benzylazanediyl)bis(hexane-6,l-diyl) diacetate (A-4)To a stirred solution of Benzyl amine (A-3) (5.0 g, 46.68 mmol) in acetonitrile (50 mL) at RT, was added DIPEA (20 mL, 116.70 mmol) followed by 6-bromohexyl acetate (A-2) (19.7 g, 88.70 mmol) in acetonitrile (50 mL) and the reaction mixture was stirred at 80 °C for 24 h (monitored by TLC). Then the reaction mixture was diluted with water (500 mL) and extracted with EtOAc (2 x 500 mL) The combined organic layer dried over anhydrous Na2SO4, filtered and the filtrate was concentrated under vacuum to afford (benzylazanediyl)bis(hexane-6,l-diyl) diacetate (A-4) (17.6 g, 44.94 mmol, 96 % yield) as colorless liquid.XH NMR (400 MHz, DMSO-d6): 6 ppm 7.40-7.20 (m, 5H), 4.10 - 3.80 (m, 4H), 3.47 (s, 2H), 2.48-2.35 (m, 4H), 2.10-1.90 (m, 6H), 1.58-1.48 (m, 4 H), 1.47-1.32 (m, 4H), 1.30-1.15 (m, 8H).Step 3: Synthesis of 6,6'-(benzylazanediyl)bis(hexan-l-ol) (A-5)To a stirred solution of benzylazanediyl-bis(hexane-6,l-diyl) diacetate (A-3) (17.6 g, 44.94 mmol) in MeOH (180 mL) at RT, was added K2CO3(18.63 g, 134.8 mmol) and the reaction mixture was stirred for 2 h (monitored by TLC). Then the reaction mixture was diluted with water (500 mL) and extracted with EtOAc (2 x 500 mL), dried over Na2SO4, filtered and the filtrate was concentrated under reduced pressure. The resulting crude compound was purified by Combi-flash column chromatography (40 g silica column) gradient elution of 10-100 % EtOAc in hexane. The resulting fractions containing desired product were combined and concentrated to afford 6,6'-(benzylazanediyl)bis(hexan-l-ol) (A-5) (12 g, 39.02 mmol, 87 % yield) as colorless liquid.XH NMR (400 MHz, DMSO-dg): 6 ppm 7.31-7.27 (m, 4H),7.23-7.19 (m, 1H), 4.33-4.30 (m, 2H), 3.48 (s, 2H), 3.37-3.32 (m, 4H), 2.32 (t, J = 6.8 Hz, 4H), 1.48-1.32 (m, 8H), 1.31-1.22 (m, 8H).Step 4: Synthesis of 6-(benzyl (6-hydroxyhexyl)amino)hexyl 2-hexyldecanoate (A-7)To a stirred solution of 2-hexyldecanoic acid (A-6) (800 mg, 0.97 mmol) in DCM (15 mL) at 0 °C, was added EDC.HCI (0.89 g, 4.66 mmol), DMAP (122 mg, 1.24 mmol) followed by 6,6'- (benzylazanediyl)bis(hexan-l-ol) (A-5) (2.1 g, 6.86 mmol) and the reaction mixture was warmed to stir at RT for 16 h (monitored by TLC). Then the reaction mixture was quenched with water (100 mL) and extracted with EtOAc (2 x 100 mL), dried over NajSCU, filtered and the filtrate was concentrated under reduced pressure. The resulting crude compound was purified by Combi-flash column chromatography (24 g silica column) gradient elution of 1-30 % EtOAc in hexane. The resulting fractions containing desired product were combined and concentrated to afford 6-(benzyl (6- hydroxyhexyl)amino)hexyl 2-hexyldecanoate (A-7) (930 mg, 1.70 mmol, 55 % yield) as a colorless liquid.TH NMR (400 MHz, DMSO-d6) : 6 ppm 7.34-7.25 (m, 4H), 7.23-7.19 (m, 1H), 4.32 (t, J = 4.8 Hz, 1H), 4.02-4.97 (m, 2H), 3.47 (s, 2H), 3.32-3.24 (m, 2H), 2.38-2.28 (m, 2H), 2.37-2.22 (m, 1H), 1.60-1.32 (m, 12H), 1.30-1.10 (m, 30 H), 0.84 (t, J = 6 Hz, 6H).Step 5: Synthesis of 6-(benzyl(6-((2-hexylnonanoyl)oxy)hexyl)amino)hexyl 2-hexyldecanoate (A-9)To a stirred solution of 2-hexylnonanoic acid (A-8) (Synthesis described in step-7) (0.53 g, 2.21 mmol) in DCM (15 mL) at 0 °C was added EDC.HCI (0.49 g, 2.55 mmol), DMAP (0.041 g, 0.34 mmol) followed by 6-(benzyl(6-hydroxyhexyl)amino)hexyl 2-hexyldecanoate (A-7) (0.930 g, 1.70 mmol) and the reaction mixture was stirred at RT for 16 h (monitored by TLC). Then the reaction mixture was quenched with water (100 mL) and extracted with EtOAc (2 x 100 mL), dried over Na2SO4, filtered and the filtrate was concentrated under reduced pressure. The resulting crude compound was purified by Combi-flash column chromatography (24 g silica column) gradient elution of 1-6 % EtOAc in hexane. The resulting fractions containing desired product were combined and concentrated to afford 6- (benzyl(6-((2-hexylnonanoyl)oxy)hexyl)amino)hexyl 2-hexyldecanoate (A-9) (0.860 g, 1.11 mmol, 65 % yield) as colorless liquid.XH NMR (400 MHz, DMSO-d6): 6 ppm 7.30-7.22 (m, 5H), 4.04 (t, J = 6.4 Hz, 4H), 3.53 (s, 2H), 2.38 (t, J = 7.2 Hz, 4H), 2.32-2.27 (m, 2H), 1.68-1.53 (m, 9 H), 1.52-1.39 (m, 8 H), 1.38- 1.20 (m, 45 H), 0.88-0.85 (m, 12 H).Step 6: Synthesis of 6-((6-((2-hexylnonanoyl)oxy)hexyl)amino)hexyl 2-hexyldecanoate (Lipid A)To a stirred solution of 6-(benzyl(6-((2-hexylnonanoyl)oxy)hexyl)amino)hexyl 2-hexyldecanoate (A-9) (0.860 g, 1.11 mmol) in EtOAc (10 mL) at RT, was added 10% Pd-C (200 mg) and stirred under hydrogen atmosphere for 16 h (monitored by TLC). Then the reaction mixture was diluted with EtOAc (50 mL) and filtered through celite bed then filtrate was evaporated under reduced pressure to afford 6-((6- ((2-hexylnonanoyl)oxy)hexyl)amino)hexyl 2-hexyldecanoate (Lipid A) (0.600 g, 0.88 mmol, 79 % yield) as a colorless syrup.!H NMR (400 MHz, DMSO-d6): 6 ppm 4.06 (t, J =6.8 Hz, 4H), 2.63 (t, J = 7.6 Hz, 4H), 2.31-2.28 (m, 2H), 1.70-1.50 (m, 12 H), 1.49-1.15 (m, 50 H), 0.89-0.85 (m, 12 H).Step 7: Synthesis of 2-hexylnonanoic acid (A-8)The freshly prepared LDA (18.64 mL, 2M in THF, 37.28 mmol) was slowly added to a solution of decanoic acid (A-10) (5.0 g, 31.59 mmol) and NaH (60 w / w% mineral oil suspension, 1.49 g, 37.28 mmol) in THF (80 mL) at 0 °C and stirred at RT for 30 min. Then, to the above solution was added iodohexane (5.52 mL, 37.28 mmol) and the reaction mixture was stirred at 45°C for 6 h (monitored by TLC). Then the reaction mixture was quenched with IN HCI (50 mL) and extracted with EtOAc (2 x 250mL). The combined organic layer was washed with water (250 mL), dried over Na2SO4, filtered and the filtrate was concentrated under reduced pressure. The resulting crude compound was purified by Combi-flash column chromatography (40 g silica column) gradient elution of 1-3 % EtOAc in hexane. The resulting fractions containing desired product were combined and concentrated to afford 2- hexylnonanoic acid (A-8) (3 g, 12.37 mmol, 39 % yield) as colorless liquid.1H NMR (400 MHz, DMSO- d6): 6 ppm 12.02 (s, 1H), 2.18-2.16 (m, 1H), 1.55-1.32 (m, 4H), 1.32-1.02 (m, 18H), 0.86-0.83 (m, 6H).Synthesis of 3-((4,4-bis(octyloxy)butanoyl)oxy)-2-(hydroxymethyl)propyl (9Z,12Z)-octadeca-9,12- dienoate (Lipid-B)Synthetic Scheme:Step 1: Synthesis of 4,4-diethoxybutanenitrile (B-2)To a stirred solution of 3-chloro-l,l-diethoxypropane (B-l) (25 g, 150 mmol) in DMSO (250 mL) was added TBAI (5.54 g, 15.00 mmol) followed by NaCN (36.8 g, 750 mmol) at RT and the reaction was heated to stir at 60 °C for 16 h (monitored by TLC). Then the reaction mixture was diluted with water (1000 mL) and extracted with ethyl acetate (2 xlOOO mL). The combined organic layers were washed with water (3 x 500 mL) followed by brine solution (500 mL) and dried over anhydrous sodium sulphate, filtered and concentrated to obtain the crude compound as colorless liquid. The resulting crude was purified by Combi-flash (80 g silica gel column) with gradient elution of 1-5% EtOAc in hexane to afford 4,4-diethoxybutanenitrile (B-2) (20 g, 127 mmol, 85 % yield) as a colorless liquid and which was confirmed by1H-NMR.TH NMR (400 MHz, CDCI3): 6 ppm 4.58 (t, J = 5.2, 1H), 3.72-3.64 (m, 2H), 3.56-3.48 (m, 2H), 2.43 (t, J = 7.6 Hz, 2H), 1.97-1.92 (m, 2H), 1.22 (t, J = 7.2, 3H).Step 2: Synthesis of 4,4-bis(octyloxy)butanenitrile (B-3)To a mixture of 4,4-diethoxybutanenitrile (B-2) (1.0 g, 6.36 mmol) and octan-l-ol (B-3) (2.485 g, 19.08 mmol) in sealed tube at RT, was added PPTS (0.799 g, 3.18 mmol). The reaction mixture was stirred at120 °C for 16 h (monitored byTLC). The reaction mixture was diluted with water (30 mL) and extracted with EtOAc (2 x 50 mL). The combined organic layers were washed with brine solution (30 mL) dried over anhydrous Na2SO4, filtered and the filtrate was concentrated under vacuum. The resulting crude material was purified by Combi-flash (40 g silica column) with gradient elution of 5% ethyl acetate in hexane. The resulting fractions containing desired product were combined and concentrated to afford 4,4-bis(octyloxy)butanenitrile (B-4) (700 mg, 2.150 mmol, 33.8 % yield) as colorless liquid. MS (ESMS): m / z 326.3 [M+H]+,TH NMR (400 MHz, CDCI3): 6 ppm 4.55 (t, J = 5.2 Hz, 1H), 3.62-3.57 (m, 2H), 3.45- 3.40 (m, 2H), 2.42 (t, J = 8.0 Hz, 2H), 1.97-1.91 (m, 2H), 1.60-1.53 (m, 4H), 1.35-1.27 (m, 20H), 0.90- 0.86 (m, 6H).Step 3: Synthesis of 4,4-bis(octyloxy)butanoic acid (B-5)To a stirred solution of 4,4-bis(octyloxy)butanenitrile (B-4) (3 g, 9.22 mmol) in EtOH (30 mL) and water (30 mL) was added potassium hydroxide (2.59 g, 46.1 mmol) and the reaction heated to stir at 110 °C for 16 h (monitored by TLC). The Reaction mixture was cooled to RT, diluted with water (100 mL) and acidified with 2N HCI aqueous solution to pH ~3 and extracted with Ethyl acetate (2 x 100 mL). The combined organic layer was washed with brine solution and dried over anhydrous sodium sulphate and concentrated to afford 4,4-bis(octyloxy)butanoic acid (B-5) (3 g, 8.71 mmol, 94 % yield) as colorless liquid and which was confirmed by1H-NMR.XH NMR (400 MHz, DMSO-dg): 6 ppm 12.07 (bs, 1H), 4.44 (t, J = 8.0 Hz, 1H), 3.50-3.45 (m, 2H), 3.37-3.31 (m, 2H), 2.21 (t, J = 8.0 Hz, 2H), 1.74-1.69 (m, 2H), 1.48-1.43 (m, 4H), 1.34-1.20 (m, 20H), 0.87-0.83 (m, 6H).Step 4: Synthesis of 3-((4,4-bis(octyloxy)butanoyl)oxy)-2-(hydroxymethyl)propyl (9Z,12Z)-octadeca- 9,12-dienoate (Lipid-B)To a stirred solution of 4,4-bis(octyloxy)butanoic acid (B-5) (3 g, 8.71 mmol)) in DCM (30 ml)) was added DIPEA (1.688 g, 13.06 mmol) and DMAP (0.213 g, 1.741 mmol) at 0 °C, followed by addition of 3-hydroxy-2-(hydroxymethyl)propyl (9Z,12Z)-octadeca-9,12-dienoate (B-6) (3.85 g, 10.45 mmol) and then EDC. HCI (2.497 g, 13.06 mmol) at 0 °C. the reaction was stirred at RT for 16 h (monitored by TLC). Then the reaction mixture was diluted with water (200.0 mL) and extracted with DCM (2X200 mL). the combined organic layers were washed with brine solution and dried over anhydrous sodium sulphate and concentrated to obtain crude compound as colorless liquid. The Crude was purified by combi flash chromatography with gradient elution of 30% EtOAc in Hexane to afford 3-((4,4- bis(octyloxy)butanoyl)oxy)-2-(hydroxymethyl)propyl(9Z,12Z)-octadeca-9,12-dienoate (Lipid-B) (2.5 g, 3.60 mmol, 41.3 % yield) as colorless liquid.XH NMR (400 MHz, CDCI3): 6 ppm 5.39-5.31 (m, 4H), 4.48 (t, J = 8.0 Hz, 1H), 4.19-4.16 (m, 4H), 3.63-3.55 (m, 4H), 3.43-3.38 (m, 2H), 2.77 (t, J = 6.8 Hz, 1H), 2.39 (t, J = 8.0 Hz, 2H), 2.32 (t, J = 7.6 Hz, 1H), 2.30-2.15 (m, 2H), 2.08-2.00 (m, 4H), 1.94-1.92 (m, 2H), 1.57- 1.53 (m, 5H), 1.33-1.27 (m, 36H), 0.90-0.86 (m, 9H).Step 5: Synthesis of 3-hydroxy-2-(hydroxymethyl)propyl (9Z,12Z)-octadeca-9,12-Dienoate (B-3) and 2-(hydroxymethyl)propane-l,3-diyl(9Z,9'Z,12Z,12'Z)-bis(octadeca-9,12-dienoate) (B-4)To a stirred solution of (9Z,12Z)-octadeca-9,12-dienoic acid (B-7) (20 g, 71.3 mmol) in DCM (200 ml) was added DIPEA (18.68 ml, 107 mmol) and DMAP (2.61 g, 21.39 mmol) at 0 °C, followed by 2- (hydroxymethyl)propane-l,3-diol (9.08 g, 86 mmol) then EDC (20.51 g, 107 mmol) at 0 °C and the reaction was stirred at RT for 16 h (monitored by TLC). Then the reaction mixture was diluted with water (500.0 mL) and extracted with DCM (2X500 mL). the combined organic layers were washed with brine solution and dried over anhydrous sodium sulphate, and concentrated to obtain crudecompound as light yellow color liquid. The Crude was purified by combi flash chromatography with gradient elution of 5% - 10% EtOAc in Hexane to afford 2-(hydroxymethyl)propane-l,3-diyl (9Z,9'Z,12Z,12'Z)-bis(octadeca-9,12-dienoate) (B-8) (4 g, 6.34 mmol, 8.89 % yield) & 3-hydroxy-2- (hydroxymethyl)propyl (9Z,12Z)-octadeca-9,12-dienoate (B-6) (8.5 g, 23.06 mmol, 32.3 % yield) as colorless liquid.TH NMR (400 MHz, DMSO-d6): 6 ppm 5.36-5.28 (m, 4H), 4.48 (t, J = 8.0 Hz, 2H), 4.00 (d, J = 6.4 Hz, 2H), 3.44-3.33 (m, 4H), 2.73 (t, J = 8.0 Hz, 2H), 2.26 (t, J = 8.0 Hz, 2H), 2.00 (t, J = 8.0 Hz, 4H), 1.82-1.79 (m, 1H), 1.52-1.48 (m, 2H), 1.31-1.25 (m, 14H), 0.87-0.84 (m, 3H).Synthesis of azanediylbis(hexane-6,l-diyl) bis(2-hexyldecanoate (Lipid C)Step 1: Synthesis of 7-bromoheptyl 2-hexyldecanoate (C-3)To a stirred solution of 2-hexylnonanoic acid (C-2) (34 g, 132 mmol) in DCM (400 mL) at 0° C, was added EDC (31.65 g, 165 mmol), DMAP (2.6 g, 22 mmol) followed by 6-bromohexan-l-ol (C-l) (34 g, 132 mmol) and the reaction mixture was warmed to stir at RT for 16 h (monitored by TLC). Then the reaction mixture was quenched with water (100 mL) and extracted with EtOAc (2 x 250 mL). The combined organic layer was dried over anhydrous Na2SO4, filtered and the filtrate was concentrated under vacuum. The resulting crude material was purified by Combi-flash (120 g silica column) with gradient elution of 1-5% EtOAc in hexane. The resulting fractions containing desired product were combined and concentrated to afford 7-bromoheptyl 2-hexyldecanoate (C-3) (22 g, 52 mmol, 47%) as a colorless liquid and which was confirmed byXH NMR.XH NMR (400 MHz, CDCh): 6 ppm 4.07 (t, J = 6.4 Hz, 2H), 3.48 (t, J = 6.8 Hz, 2H), 2.33-2.27 (m, 1H), 1.90-1.83 (m, 2H), 1.68-1.58 (m, 4H), 1.47-1.35 (m, 6H), 1.34-1.19 (m, 20H), 0.87 (t, J = 6.4 Hz, 6H).Step 2: Synthesis of (benzylazanediyl)bis(hexane-6,l-diyl) bis(2-hexyldecanoate) (C-5)To a stirred solution of phenyl methanamine (C-4) (2.8 g, 26.16 mmol, 1.0 eq.) in ACN (220 mL) at RT, was added DIPEA (11.26 ml, 65.42 mmol) followed by 6-bromohexyl 2-hexyldecanoate (C-3) (22 g, 57.5 mmol) and the reaction mixture was warmed to stir at 80 °C for 48 h (monitored by TLC). Thenthe reaction mixture was cooled to RT, diluted with water (250 mL) and extracted with EtOAc (2 x 250 mL). The organic layer was dried over anhydrous NajSCU, filtered and the filtrate was concentrated under vacuum. The resulting crude material was purified by Combi-flash (40 g silica column) with gradient elution of 1-7% EtOAc in hexane. The resulting fractions containing desired product were combined and concentrated to afford (benzylazanediyl)bis(hexane-6,l-diyl) bis(2-hexyldecanoate) (C- 5) (9 g, 11.64 mmol, 45%) as a colorless liquid and which was confirmed by1H NMR.1H NMR (400 MHz, CDCI3): 6 ppm 7.30-7.29 9 (m, 4H), 7.26-7.16 (m, 1H), 4.03 (t, J = 6.8 Hz, 4H), 3.52 (s, 2H), 2.38 (t, J = 7.2 Hz, 4H), 2.31-2.28 (m, 2H), 1.64-1.58 (m, 4H), 1.55-1.36 (m, 8H), 1.34-1.17 (m, 52H), 0.87 (t, J = 6.6 Hz, 12H).Step 3: Synthesis of azanediylbis(hexane-6,l-diyl) bis(2-hexyldecanoate) (Lipid C)To a stirred solution of 6-(benzyl(6-((2-hexylnonanoyl)oxy)hexyl)amino)hexyl 2-hexyldecanoate (C-5) (9 g, 11.64 mmol) in EtOAc (100 mL) was added 10% Pd-C (3 g) and then reaction mixture stirred at RT for 16 h (monitored by TLC). Then the reaction mixture was filtered through celite bed, washed with EtOAc (500 mL) and the filtrate was concentrated under reduced pressure to afford azanediylbis(hexane-6,l-diyl) bis(2-hexyldecanoate) (Lipid C) (7 g, 10.08 mmol, 88%) as a colorless syrup and which was confirmed by 1H-NMR.XH NMR (400 MHz, CDCI3): 6 ppm 4.06 (t, J = 6.8 Hz, 4H), 2.58 (t, J = 7.2 Hz, 4H), 2.32-2.28 (m, 2H), 1.64-1.32 (m, 24H), 1.31-1.15 (m, 42H), 0.87 (t, J = 6.4 Hz, 12H).Synthesis of 5-((3-(2-aminoethoxy)propyl)(5-((2-hexyldecanoyl)oxy)pentyl)amino)pentyl 4- hexyldecanoate (Lipid E)Synthetic Scheme:Step 1: Synthesis of benzyl (3-bromopropyl)carbamate (E-3)To a stirred solution of 3-bromopropan-l-amine hydrochloride (E-l) (20 g, 115 mmol) in EtOAc (800 mL) at 0 °C, was added a solution NajCOs (88 g, 830 mmol) in water (400 mL) and followed by benzyl chloroformate (E-2) (19.56 g, 115 mmol) and the reaction mixture was warmed to stir at RT for 16 h (monitored by TLC). Then the reaction mixture was diluted with EtOAc (200 mL) washed with water (400 mL) followed by brine solution (400 mL). The organic layer was dried over Na2SO4, filtered and the filtrate was concentrated under reduced pressure to afford benzyl (3-bromopropyl)carbamate (E- 3) (14 g, 51.4 mmol, 45% yield) as a pale-yellow liquid. 1H NMR (400 MHz, CDCh) = 6 ppm 7.50-7.30 (m, 5H), 5.09 (s, 2H), 5.00-4.72 (m, 1H), 3.44 (t, J = 6.4 Hz, 2H), 3.41-3.28 (m, 2H), 2.16-2.02 (m, 2H).Step 2: Synthesis of tert-butyl (2-(3-(((benzyloxy)carbonyl)amino)propoxy)ethyl)carbamate (E-5)To a stirred solution of benzyl (3-bromopropyl)carbamate (E-3) (14 g, 51.4 mmol) in DCM (320 mL) at 0 °C, was added aqueous sodium hydroxide (320 mL, 51.4 mmol), tert-butyl (2- hydroxyethyl)carbamate (E-4) (10.78 g, 66.9 mmol) and followed byTBAI (19.00 g, 51.4 mmol) and the reaction mixture was warmed to stir at RT for 16 h (monitored by TLC). Then the reaction mixture was diluted with DCM (400 mL) washed with water (500 mL) followed by brine solution (500 mL). The organic layer was dried over Na2SO4, filtered and the filtrate was concentrated under reduced pressure. The resulting crude compound was purified by Combi-flash (80 g silica column) with gradient elution of 10% EtOAc in Hexane. The resulting fractions containing desired product were combined and concentrated to afford tert-butyl (2-(3-(((benzyloxy)carbonyl)amino)propoxy) ethylcarbamate (E- 5) (6.5 g, 16.39 mmol, 31.9 % yield) as a pale brown liquid. 1H NMR (400 MHz, DMSO-dg) = 6 ppm 7.50-7.31 (m, 5H), 7.30-7.20 (m, 1H), 6.90-6.70 (m, 1H), 5.00 (s, 2H), 3.42-3.36 (m, 2H), 3.34-3.28 (m, 2H), 3.12-2.98 (m, 4H), 1.68-1.56 (m, 2H), 1.36 (s, 9H).Step 3: Synthesis of tert-butyl (2-(3-aminopropoxy)ethyl)carbamate (E-6)To a stirred solution of tert-butyl (2-(3-(((benzyloxy)carbonyl)amino)propoxy)ethyl)carbamate (E-5) (6.5 g, 18.44 mmol) in THF (60 mL) at RT, was added ammonia solution in methanol (7.0 M) (60 ml,420 mmol) and followed by Pd-C (3 g, 2.82 mmol) and the reaction mixture was stirred under H2balloon at same temperature for 48 h (monitored by TLC). Then the reaction mixture was directly filtered through a celite pad using MeOH (200 mL) and the filtrate was concentrated under reduced pressure to afford tert-butyl (2-(3-aminopropoxy)ethyl)carbamate (E-6) (4.0 g, 12.33 mmol, 66.9 % yield) as a pale brown liquid. 1H NMR (400 MHz, CDCU) = 6 ppm 5.20-4.86 (bs, 1H), 3.53 (t, J = 6 Hz, 2H), 3.48 (t, J = 5.2 Hz, 2H), 3.38-3.22 (m, 2H), 2.90-2.78 (m, 2H), 1.84-1.78 (m, 2H), 1.42 (s, 9H).Step 4: Synthesis of 2,2-dimethyl-4-oxo-3,8-dioxa-5,12-diazaheptadecan-17-yl 2-hexyldecanoate (E- 8)To a stirred solution of 5-oxopentyl 2-hexyldecanoate (E-6) (3.4 g, 9.98 mmol) in DCM (50 mL) at 0 °C, was added tert-butyl (2-(3-aminopropoxy)ethyl)carbamate (E-7) (synthesis described in step 8) (2.179 g, 9.98 mmol) and followed by sodium triacetoxyborohydride (4.23 g, 19.97 mmol) and the reaction mixture was warmed to stir at RT for 24 h (monitored by TLC). Then the reaction mixture was diluted with DCM (50 mL) washed with water (70 mL), aqueous NaHCO3solution (70 mL) followed by brine solution (70 mL). The organic layer was dried over Na2SO4, filtered and the filtrate was concentrated under reduced pressure. The resulting crude compound was purified by Combi-flash (40 g silica column) with gradient elution of 20-100% EtOAc in Hexane followed by 0-20% MeOH in DCM. The resulting fractions containing desired product were combined and concentrated to afford 2,2- dimethyl-4-oxo-3,8-dioxa-5,12-diazaheptadecan-17-yl 2-hexyldecanoate (E-8) (1.6 g, 2.95 mmol, 29.5 % yield) as a pale-yellow thick liquid. 1H NMR (400 MHz, CDCI3) = 6 ppm 5.47 (bs, 1H), 4.06 (t, J = 6.8 Hz, 2H), 3.75-3.60 (m, 4H), 3.58-3.53 (m, 3H), 3.52-3.44 (m, 2H), 3.36-3.24 (m, 2H), 2.84 (t, J = 6.4 Hz, 2H), 2.71 (t, J = 7.6 Hz, 2H), 2.36-2.24 (m, 1H), 1.99 (s, 2H), 1.85 (t, J = 6.0 Hz, 2H), 1.72-1.50 (m, 6H), 1.51-1.35 (m, 9H), 1.34-1.14 (m, 18H), 0.88 (t, J = 6.4 Hz, 6H).Step 5: Synthesis of 12-(5-((2-hexyldecanoyl)oxy)pentyl)-2,2-dimethyl-4-oxo-3,8-dioxa-5,12- diazaheptadecan-17-yl 4-hexyldecanoate (E-10)To a stirred solution of 2,2-dimethyl-4-oxo-3,8-dioxa-5,12-diazaheptadecan-17-yl 2-hexyldecanoate (E-8) (1.3 g, 2.395 mmol) in Acetonitrile (13 mL) at RT, was added cyclopentylmethyl ether (13 mL), potassium carbonate (1.258 g, 9.10 mmol), potassium iodide (0.080 g, 0.479 mmol) and followed by 5-bromopentyl 4-hexyldecanoate (E-9) (Synthesis described in Step-9) (1.165 g, 2.87 mmol) and the reaction mixture was warmed to stir at 80 °C for 48 h (monitored by TLC). Then the reaction mixture was cooled to RT, directly filtered using DCM (50 mL) and the filtrate was concentrated under reduced pressure. The resulting crude compound was purified by Combi-flash (40 g silica column) with gradient elution of 20-100% EtOAc in Hexane followed by 0-20% MeOH in DCM. The resulting fractions containing desired product were combined and concentrated to afford 12-(5-((2- hexyldecanoyl)oxy)pentyl)-2,2-dimethyl-4-oxo-3,8-dioxa-5,12-diazaheptadecan-17-yl-4-hexyl decanoate (E-10) (1.5 g, 1.706 mmol, 71.3 % yield) as a colorless thick liquid. HPLC-ELSD purity: 98.68%, 1H NMR (400 MHz, CDCI3) = 6 ppm 4.95 (bs, 1H), 4.15-4.00 (m, 4H), 3.50-3.40 (m, 4H), 3.30 (d, J = 4.8 Hz, 2H), 2.46 (t, J = 7.2 Hz, 2H), 2.38 (t, J = 7.2 Hz, 4H), 2.35-2.25 (m, 3H), 1.75-1.62 (m, 6H), 1.60-1.52 (m, 2H), 1.30-1.40 (m, 15H), 1.38-1.15 (m, 47H), 0.87 (t, J = 6.0 Hz, 12H).Step 6: Synthesis of 5-((3-(2-aminoethoxy)propyl)(5-((2-hexyldecanoyl)oxy)pentyl)amino)pentyl 4- hexyldecanoate (Lipid E)To a stirred solution of 12-(5-((2-hexyldecanoyl)oxy)pentyl)-2,2-dimethyl-4-oxo-3,8-dioxa-5,12- diazaheptadecan-17-yl 4-hexyldecanoate (E-10) (1.5 g, 1.729 mmol) in DCM (15 mL) at 0 °C, was addedTFA (2.0 ml, 26.0 mmol) and the reaction mixture was warmed to stir at RT for 16 h (monitored by TLC). Then the reaction mixture was diluted with DCM (20 mL) at 0 °C, quenched with aqueous NaHCO3 solution (30 mL), adjusted pH~10 and extracted with DCM (2 x 30 mL). The combined organic layer was washed with water (40 mL), aqueous NaHCO3 solution (40 mL) followed by brine solution (50 mL). The organic layer was dried over Na2SO4, filtered and the filtrate was concentrated under reduced pressure to afford 5-((3-(2-aminoethoxy)propyl)(5-((2-hexyldecanoyl)oxy)pentyl) amino)pentyl 4-hexyldecanoate (Lipid E) (1.3 g, 1.694 mmol, 98 % yield) as a pale yellow thick liquid. 1H NMR (400 MHz, CDCI3) = 6 ppm 4.07-4.03 (q, J = 6.8 Hz, 4H), 3.54-3.36 (m, 4H), 2.85 (t, J = 5.2 Hz, 2H), 2.54-2.46 (m, 2H), 2.40 (t, J = 7.6 Hz, 4H), 2.38-2.20 (m, 3H), 1.82-1.52 (m, 12H), 1.50-1.40 (m, 6H), 1.38-1.14 (m, 45H), 0.88 (t, J = 4.0 Hz, 12H).Step 7: Synthesis of 5-hydroxypentyl 2-hexyldecanoate (E-13)To a stirred solution of 2-hexyldecanoic acid (E-ll) (5 g, 19.50 mmol) and pentane-l,5-diol (E-12) (3.06 ml, 29.2 mmol) in DCM (50 mL) at 0 °C, was added DMAP (0.476 g, 3.90 mmol) followed by EDC (5.61 g, 29.2 mmol) and the reaction mixture was warmed to stir at RT for 16 h (monitored by TLC). Then the reaction mixture was diluted with DCM (50 mL) washed with water (70 mL), aqueous NaHCO3 solution (70 mL) followed by brine solution (70 mL), dried over Na2SO4, filtered and filtrate was concentrated under reduced pressure. The resulting crude compound was purified by Combi-flash column chromatography (40 g silica column) with gradient elution of 20-30% EtOAc in Hexane. The desired product containing fractions were combined and concentrated under reduced pressure to afford 5-hydroxypentyl 2-hexyldecanoate (E-13) (4 g, 11.64 mmol, 59.7% yield) as colorless liquid. HPLC-ELSD purity: 99.65%, 1H NMR (400 MHz, CDCI3): 6 ppm 4.08 (t, J = 6.4 Hz, 2H), 3.68-3.63 (q, J = 6.4 Hz, 2H), 2.38-2.22 (m, 1H), 1.74-1.60 (m, 4H), 1.58-1.53 (m, 1H), 1.52-1.36 (m, 4H), 1.36-1.26 (m, 22H), 087 (t, J = 5.6 Hz, 6H).Step 8: Synthesis of 5-oxopentyl 2-hexyldecanoate (E-7)To a stirred solution of 5-hydroxypentyl 2-hexyldecanoate (E-13) (4 g, 11.68 mmol) and TEMPO (0.365 g, 2.335 mmol) in DCM (40 mL) at 0 °C, a mixture of saturated aqueous sodium bicarbonate (11 ml, 11.68 mmol) and sodium hypochlorite (11 ml, 178 mmol) and the reaction mixture was warmed to stir at RT for 16 h (monitored by TLC). Then the reaction mixture was diluted with DCM (50 mL) washed with Hypo solution (70 mL), aqueous NaHCOs solution (70 mL) followed by brine solution (70 mL). The organic layer was dried over Na2SO4, filtered and the filtrate was concentrated under reduced pressure to afford 5-oxopentyl 2-hexyldecanoate (E-7) (3.9 g, 11.45 mmol, 98 % yield) as a red oil. 1H NMR (400 MHz, CDCI3) = 6 ppm 9.81-9.76 (m, 1H), 4.08 (t, J = 6.0 Hz, 2H), 2.54-2.44 (m, 2H), 2.38-2.24 (m, 1H), 1.82-1.64 (m, 4H), 1.62-1.50 (m, 2H), 1.52-1.38 (m, 2H), 1.36-1.18 (m, 20H), 0.87 (t, J = 6.0 Hz, 6H).Step 9: Synthesis of 5-bromopentyl 4-hexyldecanoate (E-9)To a stirred solution of 4-hexyldecanoic acid (E-14) (Synthesis described in Lipid G) (6.3 g, 24.57 mmol) and 5-bromopentan-l-ol (E-15) (6.16 g, 36.9 mmol) in DCM (70 mL) at 0 °C, was added DMAP (0.600 g, 4.91 mmol) followed by EDC (7.06 g, 36.9 mmol) and the reaction mixture was warmed to stir at RT for 16 h (monitored by TLC). Then the reaction mixture was diluted with DCM (70 mL) washed with water (100 mL), brine solution (100 mL), dried over Na2SO4, filtered and filtrate was concentrated under reduced pressure. The resulting crude compound was purified by Combi-flash column chromatography (80 g silica column) with gradient elution of 5-10% EtOAc in Hexane. The desired product containing fractions were combined and concentrated under reduced pressure to afford 5-bromopentyl 4-hexyldecanoate (E-9) (3.5 g, 8.63 mmol, 35.1 % yield) as yellow syrup. 1H NMR (400 MHz, CDCI3): 6 ppm 4.07 (t, J = 6.4 Hz, 2H), 3.41 (t, J = 6.4 Hz, 2H), 1. 1 (t, J = 8 Hz, 2H), 1.98-1.84 (m, 2H), 1.72-1.63 (m, 2H), 1.62-1.56 (m, 2H), 1.54-1.48 (m, 2H), 1.38-1.14 (m, 21H), 0.88 (t, J = 6.8 Hz, 6H).Synthesis of 7-((3-aminopropyl)(7-((2-hexyldecanoyl)oxy)heptyl)amino)heptyl 4-hexyldecanoate (Lipid F)Synthetic Scheme:Step 1: Synthesis of 7-bromoheptyl 2-hexyldecanoate (F-3)To a stirred solution of 2-hexyldecanoic acid (F-l) (10 g, 39.0 mmol) and 7-bromoheptan-l-ol (F-2) (6.00 ml, 39.0 mmol) in DCM (100 mL) were added EDC (11.21 g, 58.5 mmol) and DMAP (0.953 g, 7.80 mmol) at 0 °C. The reaction mixture was stirred at RT for 16 h (monitored byTLC). The reaction mixture was quenched with water (100 mL) and extracted with DCM (2 x 100 mL). The combined organic layers were washed with brine solution (200 mL) and dried over anhydrous Na2SO4, filtered and the filtrate was concentrated under vacuum. The resulting crude material was purified by Combi-flash (80g silica column) with gradient elution of 10-20% EtOAc in hexane. The resulting fractions containing desired product were combined and concentrated to afford 7-bromoheptyl 2-hexyldecanoate (F-3) (14 g, 32.3 mmol, 83 % yield) as a colorless liquid and which was confirmed by1H-NMR.XH NMR (400 MHz, CDCI3): 6 ppm 4.06 (t, J = 6.4 Hz, 2H), 3.40 (t, J = 6.8 Hz, 2H), 2.33-2.28 (m, 1H), 1.89-1.82 (m, 2H), 1.68-1.58 (m, 4H), 1.46-1.35 (m, 8H), 1.32-1.18 (m, 20H), 0.87 (t, J = 6.6 Hz, 6H).Step 2: 7-((3-((tert-butoxycarbonyl)amino)propyl)amino)heptyl 2-hexyldecanoate (F-5)To a stirred solution of tert-butyl (3-aminopropyl)carbamate (F-4) (7.23 g, 41.5 mmol) in Ethanol (60 mL) was added 7- bromoheptyl 2-hexyldecanoate (F-3) (6 g, 13.84 mmol) at RT. The reaction mixture heated to stir at 65 °C for 36 h (monitored by TLC). Then the reaction mixture was concentrated under vacuum. The resulting crude material was purified by Combi-flash (80 g silica column) with gradient elution of 20-100% EtOAc in hexane followed by 0-10% MeOH in DCM. The resulting fractions containing desired product were combined and concentrated to afford 7-((3-((tert- butoxycarbonyl)amino)-propyl)amino)heptyl 2-hexyldecanoate (F-5) (2.2 g, 4.18 mmol, 30.2% yield) as a pale brown thick liquid and which was confirmed by 1H-NMR.XH NMR (400 MHz, CDCI3): 6 ppm5.07 (t, J = 5.6 Hz, 1H), 4.04 (t, J = 6.8 Hz, 2H), 3.34 (q, J = 6.4 Hz, 2H), 2.99 (t, J = 5.6 Hz, 2H), 2.93 (t, J = 7.8 Hz, 2H), 2.32-2.28 (m, 1H), 2.16-2.13 (m, 2H), 1.91-1.85 (m, 2H), 1.68-1.60 (m, 2H), 1.56-1.51 (m, 2H), 1.48-1.34 (m, 18H), 1.32-1.18 (m, 20H), 0.87 (t, J = 6.8 Hz, 6H).Step 3: 7-((3-((tert-butoxycarbonyl)amino)propyl)(7-((2-hexyldecanoyl)oxy)heptyl)amino)-heptyl 4- hexyldecanoate (F-7)To a stirred solution of 7-((3-((tert-butoxycarbonyl)amino)propyl)amino)heptyl 2-hexyldecanoate (F- 5) (2.5 g, 4.75 mmol) and 7-bromoheptyl 4-hexyldecanoate (F-6) (Synthesis described in step 4 of Lipid G) (3.09 g, 7.12 mmol) in acetonitrile (25 mL) at RT, were added cyclopentylmethyl ether (25 ml), potassium carbonate (2.492 g, 18.03 mmol) followed by potassium iodide (0.158 g, 0.949 mmol) and the reaction mixture was stirred at 80 °C for 48 h (monitored by TLC).The reaction mixture was filtered under vacuum and the solids were washed with DCM (70 mL) and filtrate was concentrated under vacuum. The resulting crude material was purified by Combi-flash (40 g silica column) with gradient elution of 20-100% EtOAc in hexane. The resulting fractions containing desired product were combined and concentrated to afford 7-((3-((tert-butoxycarbonyl)amino)propyl)-(7-((2- hexyldecanoyl)oxy)heptyl)amino)heptyl 4-hexyldecanoate (F-7) (2.3 g, 2.62 mmol, 55.1 % yield) as a colorless thick liquid with structure confirmed by 1H NMR.TH NMR (400 MHz, CDCI3): 6 ppm 5.65 (bs, 1H), 4.07-4.03 (m, 4H), 3.22-3.05 (m, 2H), 2.44 (t, J = 6.8 Hz, 2H), 2.36-2.24 (m, 7H), 1.67-2.59 (m, 4H), 1.57-2.52 (m, 2H), 1.48-1.39 (m, 16H), 1.38-1.28 (m, 56H), 0.89-0.86 (m, 12H).Step 4: 7-((3-aminopropyl)(7-((2-hexyldecanoyl)oxy)heptyl)amino)heptyl 4-hexyldecanoate (Lipid F)To a stirred solution of 7-((3-((tert-butoxycarbonyl)amino)propyl)(7-((2-hexyldecanoyl)oxy)heptyl) amino)heptyl 4-hexyldecanoate (F-7) (2.3 g, 2.62 mmol) in DCM (25 ml) at 0 °C, was added TFA (4.03 ml, 52.3 mmol)and the reaction mixture was warmed to stir at RT for 16 h (monitored by TLC). The reaction mixture was diluted with DCM (50 mL), cooled to 0 °C, quenched and basified with saturated aq. NaHCOs solution (70 mL) and extracted with DCM (2 x 50 mL). The combined organic layer was washed with brine solution (70 mL) and dried over NajSCU, concentrated under vacuum to afford 7- ((3-aminopropyl)(7-((2-hexyldecanoyl)oxy-heptyl)amino)heptyl 4-hexyldecanoate (Lipid F) (2.0 g, 2.57 mmol, 98 % yield) as a pale yellow gummy liquid with structure confirmed by 1H NMR.XH NMR (400 MHz, CDCI3) = 6 ppm 4.07-4.03 (m, 4H), 2.72 (t, J = 6.8 Hz, 2H), 2.44 (t, J = 7.6 Hz, 2H), 2.37 (t, J = 7.6 Hz, 4H), 2.33-2.24 (m, 3H), 1.66-1.54 (m, 12H), 1.51-1.37 (m, 10H), 1.34-1.18 (m, 47H), 0.90-0.84 (m, 12H).Synthesis of 7-((7-((2-hexyldecanoyl)oxy)heptyl)amino)heptyl 4-hexyldecanoate (Lipid G)Synthetic Scheme:Step 1: Synthesis of 7-(methoxymethylene)tridecane (G-3)To a stirred solution of tridecan-7-one (G-l) (25 g, 126 mmol) in THF (200 ml) at RT, was added KOtBu (22.63 g, 202 mmol) followed by (methoxymethyl)triphenylphosphonium chloride (G-2) (69.1 g, 202 mmol) and the reaction mixture was stirred for 16 h (monitored by TLC). Then the reaction mixture was diluted with water (500 mL) and extracted with EtOAc (2 x 500 mL). The combined organic layer was dried over anhydrous Na2SO4, filtered and concentrated under reduced pressure. The resulting crude was purified by gravity column chromatography with gradient elution of 0-1% EtOAc in hexane. The resulting fractions containing desired product were combined and concentrated to afford 7- (methoxymethylene)tridecane (G-3) (23 g, 102 mmol, 81% yield) as a colorless liquid and which was confirmed byXH NMR.TH NMR (400 MHz, CDCI3): 6 ppm 5.73 (s, 1H), 3.51 (s, 3H), 2.02 (t, J = 6.8 Hz, 2H), 1.84 (t, J = 6.4 Hz, 2H), 1.33-1.27 (m, 16 H), 0.88 (t, J = 6.8 Hz, 6H).Step 2: Synthesis of 2-hexyloctanal (G-4)To a stirred solution of 7-(methoxymethylene)tridecane (22 g, 97 mmol) (G-3) in THF (200 mL) at RT, was added 6N HCI (70 ml, 97 mmol) and the reaction was stirred at 70 °C for 16 h (monitored by TLC). The reaction mixture was diluted with water (500 mL) and extracted with EtOAc (2 x 500 mL). The combined organic layer was dried over anhydrous Na2SO4, filtered and concentrated under reduced pressure. The resulting crude was purified by Combi-flash chromatography (80 g silica column used) with gradient elution of 0-1% EtOAc in hexane. The resulting fractions containing desired product were combined and concentrated to afford 2-hexyloctanal (G-4) (18 g, 85 mmol, 87 % yield) as a colorless liquid and which was confirmed byTH NMR.TH NMR (400 MHz, CDCI3): 6 ppm 9.54 (d, J = 3.2 Hz 1H), 2.24-2.19 (m, 1H), 1.80-1.58 (m, 4H), 1.52-1.37 (m, 4H), 1.35-1.20 (m, 12H), 0.87 (t, J = 6.8 Hz, 6H).Step 3: Synthesis of ethyl (E)-4-hexyldec-2-enoate (G-6)To a stirred solution of (2-ethoxy-2-oxoethyl)triphenylphosphonium bromide (G-5) (112 g, 261 mmol) in DCM (170 mL) at 0 °C, was added K2COs (58.6 g, 424 mmol) followed by solution of 2-hexyloctanal (G-4) (18.0 g, 85 mmol) in DCM (170 mL). The reaction mixture was stirred at 45 °C for 24 h (monitored by TLC). The reaction mixture was diluted with cold water (500 mL) extracted with EtOAc (3 x 500 mL). The combined organic layer was washed with saturated brine solution (500 mL) and dried overanhydrous Na2SO4, filtered and concentrated under reduced pressure. The resulting crude was purified by Combi-flash column chromatography (80 g silica column) with gradient elution of 0-2% EtOAc in hexane. The resulting fractions containing desired product were combined and concentrated to afford ethyl (Ej-4-hexyldec-2-enoate (G-6) (12 g, 42.5 mmol, 50.1 % yield) as a light brown color liquid and which was confirmed by1H NMR.1H NMR (400 MHz, CDCh): 6 ppm 6.76-6.69 (dd, J = 9.6 Hz, 16 Hz ,1H), 5.75 (d, J = 15.6 Hz, 1H), 4.18 (q, J = 6.8 Hz, 2H), 2.13-2.08 (m, 1H), 1.46-1.36 (m, 3H), 1.35-1.28 (m, 23 H), 0.87 (t, J = 6 Hz, 6H).Step 4: Synthesis of ethyl 4-hexyldecanoate (G-7)To a stirred solution of ethyl (Ej-4-hexyldec-2-enoate (G-6) (12 g, 42.5 mmol) in EtOH (100 ml) at RT, was added Pd / C (2.0 g, 42.5 mmol) and the reaction mixture was stirred under hydrogen gas-filled balloon (1 atm) at RT for 16 h (monitored by TLC). Then the reaction mixture was filtered on celite bed and wash with ethanol (100 mL), the filtrate was concentrated under reduced pressure to afford ethyl 4-hexyldecanoate (G-7) (11.0 g, 38.7 mmol, 91 % yield) as a colorless syrup and which was confirmed byXH NMR.XH NMR (400 MHz, CDCI3): 6 ppm 4.12 (q, J = 7.2 Hz, 2H), 2.26 (t, J = 8 Hz, 2H), 1.60-1.53 (m, 4 H), 1.31-1.23 (m, 22 H), 0.88 (t, J = 4 Hz, 6H).Step 5: Synthesis of 4-hexyldecanoic acid (G-8)To a stirred solution of ethyl 4-hexyldecanoate (G-7) (11 g, 38.7 mmol) in ethanol (100 mL) at RT, was added 6N NaOH (21.91 mL, 131 mmol) and the reaction mixture stirred at 65 °C for 16 h (monitored by TLC). The reaction mixture was concentrated under reduced pressure to remove volatile solvent and residue was diluted with water (30 mL), acidified with IN HCI (pH=5) and extract with EtOAc (3 x 100 mL).The combined organic layer was washed with saturated brine solution (2 x 100 mL ) and dried over anhydrous Na2SO4, filtered and concentrated under reduced pressure to afford 4-hexyldecanoic acid (G-8) (9.0 g, 35.1 mmol, 91 % yield) as a colorless syrup and which was confirmed byXH NMR.XH NMR (400 MHz, CDCI3): 6 ppm 12.02 (bs, 1H), 2.15 (t, J = 7.6 Hz, 2H), 1.46 (t, J = 6.8 Hz, 2H), 1.30- 1.10 (m, 21 H), 0.85 (t, J = 6.4 Hz, 6H).Step 6: Synthesis of 7-bromoheptyl 4-hexyldecanoate (G-10)To a stirred solution of 4-hexyldecanoic acid (G-8) (9 g, 35.1 mmol),7-bromoheptan-l-ol (G-9) (10.27 g, 52.6 mmol) in DCM (100 mL) at 0 °C, was added DMAP (4.29 g, 35.1 mmol) followed by EDC.HCI (10.09 g, 52.6 mmol) and the reaction mixture was stirred at RT for 16 h (monitored by TLC). Then the reaction mixture was diluted with water (100 mL) and extracted with DCM (2 x 100 mL). The combined organic layer was washed with brine solution (100 mL) and dried over Na2SO4, filtered and concentrated under vacuum. The resulting crude compound was purified by Combi-flash column chromatography (80 g silica column) with gradient elution of 0-4 % EtOAc in hexane. The resulting fractions containing desired product were combined and concentrated to afford 7-bromoheptyl 4- hexyldecanoate (G-10) (10.2 g, 23.53 mmol, 67.0 % yield) as a colorless liquid and which was confirmed by1HNMR.XH NMR (400 MHz, CDCI3): 6 ppm 4.05 (t, J = 6.8 Hz, 2H), 3.41 (t, J = 6.8 Hz, 2H), 1. 1 (t, J = 8 Hz, 2H), 1.91-1.82 (m, 2H), 1.66-1.56 (m, 4H), 1.50-1.42 (m, 2H), 1.40-1.16 (m, 25H), 0.88 (t, J= 6.4 Hz, 6H).Step 7: Synthesis of 7-(benzyl(7-((2-hexyldecanoyl)oxy)heptyl)amino)heptyl 4-hexyldecanoate (G- 12)To a stirred solution of 7-(benzylamino)heptyl 2-hexyldecanoate (G-ll) (Synthesis described in step 9) (2.8 g, 6.09 mmol) in acetonitrile (20 ml) at RT, was added DIPEA (2.77 ml, 15.23 mmol) followed by 7-bromoheptyl 4-hexyldecanoate (G-10) (2.64 g, 6.09 mmol) and the reaction mixture was stirred at 80 °C for 36 h (monitored by TLC). Then the reaction mixture was concentrated under reduced pressure. The resulting crude compound was purified by combi-flash column chromatography (40 g silica column) with gradient elution of 20-30 % of EtOAc in hexane. The resulting fractions containing desired product were combined and concentrated to afford 7-(benzyl(7-((2- hexyldecanoyl)oxy)heptyl)amino)heptyl 4-hexyldecanoate (G-12) (3.0 g, 3.69 mmol, 60.6 % yield) as a pale yellow syrup and which was confirmed by1H NMR.1H NMR (400 MHz, CDCU): 6 ppm 7.31-7.27 (m, 4 H), 7.23-7.20 (m, 1 H), 4.04 (t, J = 6.4 Hz, 4H), 3.52 (s, 2H), 2.38 (t, J = 6.8 Hz, 4H), 2.34-2.24 (m, 3H), 1.67-1.61 (m, 2H), 1.59-1.57 (m, 4H), 1.46-1.39 (m, 7H) 1.37-1.16 (m, 54 H), 0.89-0.85 (m, 12H).Step 8: Synthesis of 7 7-((7-((2-hexyldecanoyl)oxy)heptyl)amino)heptyl 4-hexyldecanoate (Lipid G)To a stirred solution of 7-(benzyl(7-((2-hexyldecanoyl)oxy)heptyl)amino)heptyl 4-hexyldecanoate (G- 12) (3.0 g, 3.69 mmol) in ethyl acetate (20 ml) at 0°C, was added Pd-C (0.650 g, 3.69 mmol) and the reaction mixture was stirred under hydrogen gas balloon (1 atm) at RT for 16 h (monitored by TLC). The reaction mixture was filtered on celite bed and wash with EtOAc (50 mL) and filtrate was evaporated under reduced pressure to afford 7-((7-((2- hexyldecanoyl)oxy)-heptyl)amino)heptyl 4- hexyldecanoate (Lipid-G) ( 2.5 g, 3.46 mmol, 94 % yield) as a grey syrup and which was confirmed by1HNMR.XH NMR (400 MHz, CDCI3): 6 ppm 4.05 (t, J = 6.4 Hz, 4H), 2.58 (t, J = 7.2 Hz, 4H), 2.30-2.24 (m, 3H), 1.63-1.39 (m, 14 H), 1.40-1.18 (m, 54H), 0.89-0.85 (m, 12H).Step 9: Synthesis of 7-(benzylamino)heptyl 2-hexyldecanoate (G-ll)The stirred solution of benzylamine (G-15) (7.42 g, 69.2 mmol) in ethanol (60 mL) at RT, was added 7- bromoheptyl 2-hexyldecanoate (G-13, aka F-3) (Synthesis described in Lipid F) (6.0 g, 13.84 mmol) and the reaction mixture was stirred at 65 °C for 48 h (monitored by TLC). Then the reaction mixture was cooled to RT and concentrated under reduced pressure. The resulting crude compound was purified by Combi-flash column chromatography (40 g silica column) with gradient elution of 15-20 % EtOAc in hexane. The resulting fractions containing desired product were combined and concentrated to afford 7-(benzylamino)heptyl 2-hexyldecanoate (G-ll) (5 g, 10.88 mmol, 79 % yield) as a colorless syrup. MS (ESI): m / z 460.4 [M+H]+,TH NMR (400 MHz, CDCI3): 6 ppm 7.34-7.30 (m, 4H), 7.26-7.23 (m, 1H), 4.05 (t, J = 6.8 Hz, 2H), 3.78 (s, 2H), 2.62 (t, J = 7.2 Hz 2H), 2.31-2.29 (m, 1H), 1.62-1.49 (m, 8 H), 1.34-1.28 (m, 7 H), 1.32-1.16 (m, 20H), 0.87 (t, J = 4 Hz, 6H).Synthesis of ((4-hydroxybutyl)azanediyl)bis(hexane-6,l-diyl) bis(2-hexyldecanoate) (Lipid I):Lipid ISynthetic Scheme:Step 1: Synthesis of 4-((tert-butyldimethylsilyl)oxy)butan-l-amine (1-2)To a stirred solution of 4-aminobutan-l-ol (1-1) (5 g, 56.09 mmol) in DCM (60 mL) at 0 °C, was added triethylamine (19.57 mL, 140.2 mmol) followed by tert-Butyldimethylchlorosilane (12.68 g, 84.13 mmol) and the reaction mixture was warmed to stir at RT for 16 hr (monitored by TLC). Then the reaction mixture was diluted with DCM (70 mL) and washed with water (100 mL) followed by brine solution (100 mL). The organic layer was dried over NajSCU, filtered and the filtrate was concentrated under reduced pressure. The resulting crude compound was purified by Combi-flash (40 g silica column) with gradient elution of 10-20% solvent A in solvent B (solvent A: 0.5% TEA in MeOH and solvent B: DCM). The resulting fractions containing desired product were combined and concentrated to afford 4-((tert-butyldimethylsilyl)oxy)butan-l-amine (1-2) (8 g, 39.40 mmol, 70% yield) as a pale yellow thick liquid. 1H NMR (400 MHz, DMSO-d6): 6 ppm 3.52 (t, J = 6 Hz, 2H), 2.52-2.46 (m, 4H), 1.48- 1.38 (m, 2H), 1.36-1.26 (m, 2H), 0.82 (s, 9H), -0.01 (s, 6H).Step 2: Synthesis of ((4-((tert-butyldimethylsilyl)oxy)butyl)azanediyl)bis(hexane-6,l-diyl) bis(2- hexyldecanoate) (1-4)To a stirred solution of 4-((tert-butyldimethylsilyl)oxy)butan-l-amine (1-2) (0.7 g, 3.44 mmol) in Acetonitrile (20 mL) at RT, was added DIPEA (1.503 ml, 8.60 mmol) and followed by 6-bromohexyl 2- hexyldecanoate (1-3) (synthesis described in step 4) (3.18 g, 7.57 mmol) and the reaction mixture was heated to stir at 80 °C for 48 h (monitored by TLC). Then the reaction mixture was diluted with EtOAc (50 mL), washed with water (50 mL) followed by brine solution (50 mL). The organic layer was dried over NajSCU, filtered and the filtrate was concentrated under reduced pressure. The resulting crude compound was purified by Combi-flash (40 g silica column) with gradient elution of 20-100% EtoAc in Hexane and 0-20% MeOH in DCM. The resulting fractions containing desired product were combined and concentrated to afford ((4-((tert-butyldimethylsilyl)oxy)butyl)azanediyl)-bis(hexane-6,l-diyl) bis(2-hexyldecanoate) (1-4) (2 g, 2.308 mmol, 67% yield) as a pale yellow liquid. 1H NMR (400 MHz, CDCI3): 6 ppm 4.10-4.00 (m, 4H), 3.70-3.60 (m, 2H), 2.45-2.35 (m, 6H), 2.34-2.25 (m, 2H), 1.70-1.60 (m, 4H), 1.50-1.20 (m, 62H), 1.00-0.80 (m, 21H), 0.09 (s, 6H).Step 3: Synthesis of ((4-hydroxybutyl)azanediyl)bis(hexane-6,l-diyl) bis(2-hexyldecanoate) (Lipid I)To a stirred solution of ((4-((tert-butyldimethylsilyl)oxy)butyl)azanediyl)bis(hexane-6,l-diyl) bis(2- hexyldecanoate) (1-4) (2 g, 2.271 mmol) in THF (20 mL) at 0 °C, was added TBAF (1.0 M in THF) (6.81 mL, 6.81 mmol) and the reaction mixture was warmed to stir at RT for 3 h (monitored by TLC). Then the reaction mixture was diluted with EtOAc (50 mL), washed with water (50 mL) followed by brine solution (50 mL). The organic layer was dried over NajSCU, filtered and the filtrate was concentrated under reduced pressure. The resulting crude compound was purified by Combi-flash (40 g silicacolumn) with gradient elution of 20-100% EtOAc in hexane followed by 0-10% MeOH in DCM. The resulting fractions containing desired product were combined and concentrated to afford ((4- hydroxybutyl)azanediyl)bis(hexane-6,l-diyl) bis(2-hexyldecanoate) (Lipid H) (0.445 g, 0.578 mmol, 25% yield) as a colorless thick liquid. MS (ESMS): m / z 767.7 [M+H]+, HPLC-CAD purity: 90.34 %, HPLC- ELSD purity: 99.53 %,TH NMR (400 MHz, CDCI3): 6 ppm 4.05 (t, J = 6.8 Hz, 4H), 3.64-3.48 (m, 2H), 2.57- 2.36 (m, 6H), 2.35-2.22 (m, 2H), 1.78-1.52 (m, 12H), 1.52-1.06 (m, 56H), 0.87 (t, J = 6 Hz, 12H).Step 4: Synthesis of 6-bromohexyl 2-hexyldecanoate (1-3)To a stirred solution of 6-bromohexan-l-ol (1-5) (10 g, 55.22 mmol) and 2-hexyldecanoic acid (1-6) (19.44 mL, 66.27 mmol) in DCM (150 mL) at 0 °C, was added DMAP (1.34 g, 11.04 mmol) followed by EDC (15.88 g, 82.84 mmol) and the reaction mixture was warmed to stir at RT for 16 h (monitored by TLC). Then the reaction mixture was diluted with DCM (100 mL) washed with water (100 mL), aqueous NaHCOs solution (100 mL) followed by brine solution (100 mL), dried over NajSCU, filtered and filtrate was concentrated under reduced pressure. The resulting crude compound was purified by Combi-flash column chromatography (220 g silica column) with gradient elution of 5-10% EtoAc in hexane. The desired product containing fractions were combined and concentrated under reduced pressure to afford 6-bromohexyl 2-hexyldecanoate (1-3) (12 g, 28.63 mmol, 52% yield) as colorless liquid. 1H NMR (400 MHz, CDCI3): 6 ppm 4.07 (t, J = 6.4 Hz, 2H), 3.41 (t, J = 6.4 Hz, 2H), 2.38-2.25 (m, 1H), 1.95-1.82 (m, 2H), 1.72-1.60 (m, 3H), 1.56-1.34 (m, 7H), 1.33-1.18 (m, 20H), 087 (t, J = 6.4 Hz, 6H).Synthesis of 3-((4,4-bis(octyloxy)butanoyl)oxy)-2-((((3-(diethylamino)propoxy)carbonyl)oxy) methyl)propyl (9Z,12Z)-octadeca-9,12-dienoate (Lipid J)Step-1: Synthesis of 3-((4,4-bis(octyloxy)butanoyl)oxy)-2-((((3-(diethylamino)propoxy)- carbonyl)oxy)methyl)propyl (9Z,12Z)-octadeca-9,12-dienoate (Lipid J)To a stirred solution of 3-((4,4-bis(octyloxy)butanoyl)oxy)-2-(hydroxymethyl)propyl (9Z,12Z)- octadeca-9,12-dienoate (Lipid B) (4.0 g, 5.75 mmol) in DCM (40 mL) at 0 °C, was added pyridine (0.937 mL, 11.51 mmol)), DMAP (0.705 g, 5.75 mmol) followed by 4-Nitrophenyl chloroformate (J-l) (1.392 g, 6.91 mmol) and the reaction mixture was stirred at RT for 2 h. Then to the above reaction mixture was added DIPEA (3.95 mL, 23.02 mmol) and 3-(diethylamino)propan-l-ol (3.02 g, 23.02 mmol) and stirred at RT for further 16 h (monitored by TLC). Then the reaction mixture was diluted with water (100 mL) and extracted with DCM (2 x 100 mL). The combined organic layer was washed with brine solution (50 mL) dried over anhydrous NajSC , filtered and concentrated under vacuum. The resultingcrude was purified by Combi-flash (40 g silica column) with gradient elution of 2-3% MeOH in DCM. The resulting fractions containing desired product were combined and concentrated to afford 3-((4,4- bis(octyloxy)butanoyl)oxy)-2-((((3-(diethylamino)propoxy) carbonyl)oxy)methyl)propyl (9Z,12Z)- octadeca-9,12-dienoate (Lipid J) (3.64 g, 4.26 mmol, 74.1% yield) as colorless gummy syrup. MS (ESI): m / z 852.75 [M+H]+, HPLC-CAD purity: 98.58 %, HPLC-ELSD purity: 98.93 %,TH NMR (400 MHz, CDCI3) = 6 ppm 5.39-5.29 (m, 4H), 4.48 (t, J = 6.0 Hz, 1H), 4.20-4.13 (m, 8H), 3.57-3.53 (m, 2H), 3.42-3.38 (m, 2H), 2.77 (t, J = 6.8 Hz, 2H), 2.53-2.48 (m, 6H), 2.42-2.38 (m, 3H), 2.36-2.28 (m, 2H), 2.09-2.02 (m, 4H), 1.96-1.89 (m, 2H), 1.84-1.78 (m, 2H), 1.56-1.52 (m, 4H), 1.30-1.27 (m, 36H), 1.01 (t, J = 6.8 Hz, 6H), 0.90-0.86 (m, 9H).Synthesis of 3-(diethylamino)propyl 3-(hydroxymethyl)benzoate (Core-1)Synthetic Scheme:Step 1: Synthesis of methyl 3-(hydroxymethyl)benzoate (Cl-2)To a stirred solution of methyl 3-formylbenzoate (Cl-1) (10 g, 60.91 mmol) in MeOH (150 mL) at 0 °C, was added NaBH4(1.85 g, 48.73 mmol) portion wise over 10 minutes. Then the reaction mixture was stirred at same temperature for 1 h (monitored by TLC). Then the reaction mixture was quenched with ice cold water, extracted with DCM (3 x 300 mL). The combined organic layer was dried over anhydrous Na?SO4, filtered and filtrate was concentrated under reduced pressure to afford methyl 3- (hydroxymethyl)benzoate (Cl-2) (8.5 g, 51.14 mmol, 84 % yield) as colorless syrup.XH NMR (400 MHz, CDCI3): 6 ppm 7.99 (s, 1H), 7.92-7.90 (d, J = 7.6 Hz, 1H), 7.54-7.52 (d, J = 7.6 Hz, 1H), 7.39 (t, J = 7.6 Hz, 1H), 4.68 (s, 2 H), 3.88 (s, 1H).Step 2: Synthesis of methyl 3-(((tetrahydro-2H-pyran-2-yl)oxy)methyl)benzoate (Cl-3)To a stirred solution of methyl 3-(hydroxymethyl)benzoate (Cl-2) (8.5 g, 51.14 mmol) and DHP (7.02 ml, 84.12 mmol) in DCM (90 mL), was added PPTS (0.26 mg, 1.02 mmol) and the reaction mixture was stirred at RT for 8 h (monitored by TLC). Then the reaction mixture was diluted with DCM (600 mL), washed with brine (250 mL), water (2 x 200 mL), dried over NajSC , filtered and filtrate was concentrated under reduced pressure to afford methyl 3-(((tetrahydro-2H-pyran-2- yl)oxy)methyl)benzoate (Cl-3) (11.5 g, 45.94 mmol, 90 % yield) as colorless syrup.XH NMR (400 MHz, CDCI3): 6 ppm 8.04 (s, 1H), 7.97-7.95 (d, J = 8 Hz, 1H), 7.58-7.56 (d, J = 7.6 Hz, 1H), 7.42 (t, J = 8 Hz ,1H), 4.84-4.81 (d, J = 12 Hz, 1H) , 4.73-4.72 (m, 1H), 4.55-4.52 (d, J = 12.4 Hz, 1H), 3.95-3.88 (m, 3H),3.58- 3.53 (m, 1H), 1.94-1.82 (m, 1H), 1.81-1.49 (m, 6H).Step 3: Synthesis of methyl 3-(((tetrahydro-2H-pyran-2-yl)oxy)methyl)benzoate (Cl-4)To stirred solution of methyl 3-(((tetrahydro-2H-pyran-2-yl)oxy)methyl)benzoate (Cl-3) (11.5 g, 45.94 mmol) in MeOH (300 mL) at RT was added 10% aq. KOH (103 mL, 183.78 mmol) and the reaction mixture was warmed to stirred at 70 °C for 1 h (monitored by TLC). Then the reaction mixture was cooled to room temperature, concentrated to remove MeOH and the aqueous layer was washed with EtOEt (2 x 100 mL). The aqueous layer was cooled to 0 °C and acidified saturated aq. KHSO4solution, extracted with EtOAc (2 x 300 mL). The combined organic layer was dried over anhydrous Na2SO4, filtered and filtrate was concentrated under reduced pressure to afford methyl 3-(((tetrahydro-2H- pyran-2-yl)oxy)methyl)benzoate (Cl-4) (9 g, 38.09 mmol, 83 % yield) as colorless syrup.XH NMR (400 MHz, CDCI3): 6 ppm 11.0- 9.50 (s, 1H), 8.12 (s, 1H), 8.05-8.03 (d, J = 7.4 Hz, 1H), 7.64-7.62 (d, J = 7.6 Hz, 1H), 7.47 (t, J = 8 Hz ,1 H), 4.87-4.84 (d, J = 12.4 Hz, 1H) , 4.76-4.75 (m, 1H), 4.58-4.55 (d, J = 12 Hz, 1H), 3.93-3.90 (m, 1H), 3.61-3.56 (m, 1H), 1.98-1.86 (m, 1H), 1.84-1.49 (m, 6H).Step 4: Synthesis of methyl 3-(((tetrahydro-2H-pyran-2-yl)oxy)methyl)benzoate (Cl-6)To stirred solution of 3-(diethylamino)propan-l-ol (Cl-5) (2.75 g, 20.95 mmol), 3-(((tetrahydro-2H- pyran-2- yl)oxy)methyl)benzoic acid (Cl-4) (5.94 g, 25.14 mmol) and DMAP (0.52 g, 4.19 mmol) in DCM (60 mL) at 0 °C, was added EDCI.HCI (6.03 g, 31.43 mmol) and the reaction mixture was stirred RT for 16 h (monitored by TLC). Then the reaction mixture was diluted with DCM (500 mL), washed with water (3 x 200 mL). The combined organic layer was washed with brine solution (150 mL), dried over anhydrous Na2SO4, filtered and filtrate was concentrated under reduced pressure to afford methyl 3- (((tetrahydro-2H-pyran-2-yl)oxy)methyl)benzoate (Cl-6) (6.5 g, 18.59 mmol, 89 % yield) as brown syrup.XH NMR (400 MHz, CDCI3) : 6 ppm 8.03 (s, 1H), 7.97-7.95 (d, J = 8 Hz, 1H), 7.58-7.56 (d, J = 7.6 Hz, 1H), 7.42 (t, J = 8 Hz ,1 H), 4.84-4.81 (d, J = 12.4 Hz, 1H) , 4.72 (t, J = 3.2 Hz, 1H), 4.56-4.52 (d, J = 12.4 Hz, 1H), 4.37 (t, J =6.4 Hz, 2H), 3.94-3.89 (m, 1H), 3.57-3.55 (m, 1H), 2.61-2.51 (m, 6H), 1.93-1.53 (m, 8H), 1.03 (t, J = 7.2 Hz, 6H).Step 5: Synthesis of 3-(diethylamino)propyl 3-(hydroxymethyl)benzoate (Core-1)To a stirred solution of 3-(diethylamino)propyl 3-(((tetrahydro-2H-pyran-2-yl) oxy) methyl)benzoate (Cl-6) (10 g, 28.6 mmol) in MeOH (100 mL) at 0 °C, was added aqueous 2N HCI (30 mL) and the reaction mixture was slowly warmed to stir at RT for 5 h (monitored by TLC). Then the reaction mixture was quenched & basified with saturated aq. NaHCOs solution, extracted with DCM (3 x 150 mL). The combined organic layer was dried over anhydrous Na2SO4, filtered and filtrate was concentrated under reduced pressure to afford 3-(diethylamino)propyl 3-(hydroxymethyl)benzoate (Core-1) (7.2 g, 30.1 mmol, 96 % yield) as light brown thick syrup.XH NMR (400 MHz, CDCI3) : 6 ppm 8.00 (s, 1H), 7.92-7.90 (d, J = 7.6 Hz, 1H), 7.56-7.54 (d, J = 7.2 Hz, 1H), 7.40 (t, J = 7.6 Hz ,1 H), 4.70 (s, 2H) , 3.52-3.48 (m, 1H), 2.61-2.52 (m, 6H), 1.94-1.88 (m, 2H), 1.54-1.50 (m, 2H), 1.03 (t, J = 7.2 Hz, 6H).Synthesis of 4-(diethylamino)butyl 3-(hydroxymethyl)benzoate (Core-2)Synthetic Scheme:Step 1: Synthesis of methyl 3-(hydroxymethyl)benzoate (C2-2)To a stirred solution of methyl 3-formylbenzoate (C2-1) (10.0 g, 60.9 mmol) in MeOH (100 mL) at 0 °C, was added NaBH4(1.844 g, 48.7 mmol) portion wise over a period of 10 minutes. The reaction mixture was stirred at 0 °C for 1 h (monitored by TLC). The reaction mixture was quenched with ice cold water (150 mL) at 0 °C, then extracted with DCM (3 x 200 mL). The combined organic layers was dried over Na2SO4, filtered and the filtrate was concentrated under reduced pressure to afford methyl 3- (hydroxymethyl)benzoate (C2-2) (9.0 g, 54.2 mmol, 89 % yield) as colorless syrup which was confirmed by NMR. NMR (400 MHz, DMSO-d6): 6 ppm 7.94(s, 1H), 7.83(d, 7=7.6Hz, 1H), 7.58(d, 7=7.6Hz, 1H), 7.47(t, 7=7.6Hz, 1H), 5.35(t, 7=5.6Hz, 1H), 4.56(d, 7=5.6Hz, 2H), 3.86(s, 3H).Step 2: Synthesis of methyl 3-(((tetrahydro-2H-pyran-2-yl)oxy)methyl)benzoate (C2-3)To a stirred solution of methyl 3-(hydroxymethyl)benzoate (C2-2) (9.0 g, 54.2 mmol) and tetrahydro- 2H-pyran (7.0 g, 81.0 mmol) in DCM (100 mL) at 0 °C, was added PPTS (0.272 g, 1.083 mmol). The reaction mixture was stirred at RT for 6 h (monitored by TLC). The reaction mixture was diluted with DCM (300 mL) washed with water (2 x 200 mL), brine solution (250 mL). The organic layer was dried over Na2SO4, filtered and the filtrate was concentrated under vacuum. The resulting crude compound was purified by combiflash (40 g, silica column) with gradient elution of 10-15% EtOAc in Hexane. The resulting fractions containing desired product were combined and concentrated to afford methyl 3- (((tetrahydro-2H-pyran-2-yl)oxy)methyl)benzoate (C2-3) (11.2 g, 44.7 0 mmol, 83 % yield) as colorless syrup.XH NMR (400 MHz, CDCI3): 6 ppm 8.04(s, 1H), 7.96(d, 7=7.6Hz, 1H), 7.57(d, 7=7.6Hz, 1H), 7.42(t, 7=7.6Hz, 1H), 4.82(d,7=12.4Hz, 1H), 4.72(t, 7=3.2Hz, 1H), 4.54(d,7=12.4Hz, 1H), 3.9403.88(m, 4H), 3.58- 3.53(m, 1H), 1.86-1.81(m, 1H), 1.76-1.59(m, 4H), 1.55-1.53(m, 1H).Step 3: Synthesis of 3-(((tetrahydro-2H-pyran-2-yl)oxy)methyl)benzoic acid (C2-4)To a stirred solution of methyl 3-(((tetrahydro-2H-pyran-2-yl)oxy)methyl) benzoate (C2-3) (6 g, 23.97 mmol) in MeOH (100 ml) at RT, was added 10% aq. KOH (5.0 ml, 23.97 mmol). The reaction mixture was stirred at 90 °C for 4 h (monitored by TLC). The reaction mixture was cooled to RT concentratedunder vacuum to remove MeOH, the aqueous layer was washed with diethyether (100 mL). Then the aqueous layer was cooled to 0 °C, acidified with saturated aqueous KHSO4solution which was extracted with EtOAc (2 x 200 mL) and the combined organic layer was dried over anhydrous Na2SO4, filtered and the filtrate was concentrated under reduced pressure to afford 3-(((tetrahydro-2H-pyran- 2-yl)oxy)methyl)benzoic acid (C2-4) (5.1 g, 21.590 mmol, 90% yield) as colorless syrup.1H NMR (400 MHz, DMSO-dg): 6 ppm 12.93(bs, 1H), 7.91(s, 1H), 7.86(d, 7=8Hz, 1H), 7.58(d, 7=7.6Hz, 1H), 7.48(t, 7=7.6Hz, 1H), 4.74-4.69(m, 2H), 4.51(d, 7=12.4Hz, 1H), 3.81-3.76(m, 1H), 3.49-3.47(m, 1H), 1.78- 1.66(m, 2H), 1.53-1.48(m, 4H).Step 4: Synthesis of 4-(diethylamino)butyl 3-(((tetrahydro-2H-pyran-2-yl)oxy)methyl)benzoate (C2- 6)To a stirred solution of 3-(((tetrahydro-2H-pyran-2-yl)oxy)methyl)benzoic acid (C2-4) (1.952 g, 8.26 mmol) in DCM (100 ml) at 0 °C, were added DMAP (0.210 g, 1.721 mmol), EDC.HCI (1.980 g, 10.33 mmol) followed by 4-(diethylamino)butan-l-ol (C2-5) (1.0 g, 6.88 mmol). The reaction mixture was stirred at RT for 16 h (monitored by TLC). The reaction mixture was quenched with saturated NaHCOs solution (100 mL) extracted with DCM (2 x 150 mL). The combined organic layers was washed with brine solution (100 mL) dried over Na2SO4, filtered and the filtrate was concentrated under reduced pressure. The resulting crude compound was purified by combiflash (24 g, silica column) with gradient elution of 70-80 % EtOAc in Hexane. The resulting fractions containing desired product were combined and concentrated to afford 4-(diethylamino)butyl 3-(((tetrahydro-2H-pyran-2-yl)oxy)methyl)benzoate (C2-6) (0.8 g, 2.201 mmol, 32.0 % yield) as colorless liquid.XH NMR (400 MHz, DMSOd6) = 7.93(s, 1H), 7.88(d, 7=8Hz, 1H), 7.62(d, 7=7.6Hz, 1H), 7.51(t, 7=8Hz, 1H), 4.75-4.69(m, 2H), 4.53(d, 7=12.4Hz, 1H), 4.29(t, 7=6.4Hz, 2H), 3.80-3.77(m, 1H), 3.49-3.46(m, 1H), 2.45-2.39(m, 6H), 1.73-1.66(m, 4H), 1.55- 1.46(m, 6H), 0.92(t, 7=7.2Hz, 6H).Step 5: Synthesis of 4-(diethylamino)butyl 3-(hydroxymethyl)benzoate (Core-2)To a stirred solution of 4-(diethylamino)butyl 3-(((tetrahydro-2H-pyran-2-yl)oxy)methyl) benzoate (C2-6) (0.900 g, 2.47 mmol) in methanol (20 ml) at RT, was added 2N HCI (4 ml, 2.47 mmol). The reaction mixture was stirred at RT for 4 hr (monitored by TLC). Volatile solvents were removed under reduced pressure and obtained residue was diluted with EtOAc (60 mL) and washed with saturated aqueous NaHCOs solution (2 x 50 mL). The organic layer was dried over anhydrous Na2SO4, filtered and the filtrate was concentrated under reduced pressure to afford 4-(diethylamino)butyl 3- (hydroxymethyl)benzoate (Core-2) (0.5 g, 1.790 mmol, 72.5 % yield) as colorless liquid. LCMS: (M+l):280.20,XH NMR (400 MHz, DMSOd6) = 7.93(s, 1H), 7.82(d, 7=7.6Hz, 1H), 7.57(d, 7=7.6Hz, 1H), 7.47(t, 7=7.2Hz, 1H), 5.35(t, 7=6Hz, 1H), 4.56(d, 7=5.6Hz, 2H), 4.28(t, 7=6.4Hz, 2H), 2.40-2.46(m, 6H), 1.73-1.70(m, 2H), 1.53-1.49(m, 2H), 0.94(t, 7=6.8Hz, 6H).Synthesis of 3-(methyl(2-(trityloxy)ethyl)amino)propyl 3-(hydroxymethyl)benzoate (Core-3)Synthetic Scheme:Step 1: Synthesis of methyl 3-(hydroxymethyl)benzoate (C3-2)To a stirred solution of methyl 3-(hydroxymethyl)benzoate (C3-1) (9.0 g, 54.8 mmol) in MeOH (90 mL) at 0 °C, was added NaBH4(1.659 g, 43.9 mmol) portion wise and the reaction mixture was stirred at same temperature for 1 h (monitored by TLC). Then the reaction mixture was quenched with ice cold water and extracted with DCM (3 x 200 mL). The combined organic layer was dried over Na2SO4, filtered and the filtrate was concentrated under reduced pressure. The resulting crude compound was purified by Combi-flash column chromatography (40 g silica column) gradient elution of 20-30 % EtOAc in hexane. The resulting fractions containing desired product were combined and concentrated to afford methyl 3-(hydroxymethyl)benzoate (C3-2) (8.0 g, 48.1 mmol, 88 % yield) as colorless syrup.1H NMR (400 MHz, CDCI3): 6 ppm 8.01 (s, 1H), 7.95-7.93 (d, J = 6.8 Hz, 1H), 7.57-7.55 (d, J = 6.8 Hz, 1H), 7.44-7.40 (m, 1H), 4.66 (s, 2H), 3.91 (s, 3H), 2.39 (s, 3H).Step 2: Synthesis of 3-(hydroxymethyl) benzoic acid (C3-3)To a stirred solution of methyl 3-(hydroxymethyl)benzoate (C3-2) (4.0 g, 24.07 mmol) in methanol (30 mL) at 0 °C, was added 2M sodium hydroxide solution (36.1 mL, 72.2 mmol) and the reaction mixture was stirred at RT for 16 h (monitored by TLC). Then the reaction mixture was concentrated to remove EtOH, the aqueous layer was diluted with water (50 mL) and washed with diethyl ether (3 x 60 mL). The resulting aqueous layer was acidified with IM aqueous HCI at 0 °C and extracted with EtOAc (3 x 150 mL). The combined organic layers were dried over anhydrous Na2SO4, filtered and the filtrate was concentrated under reduced pressure to afford 3-(hydroxymethyl) benzoic acid (C3-3) (2.8 g, 18.40 mmol, 76 % yield) as off-white solid.TH NMR (400 MHz, CDCI3): 6 ppm 8.09 (s, J = 8 Hz, 1H), 7.64-7.63 (d, J = 7.2 Hz,lH), 7.52-7.46 (m, 1H), 4.78 (s, 2H).Step 3: Synthesis of 3-(((tert-butyldiphenylsilyl)oxy)methyl)benzoic acid (C3-4)To a stirred solution of 3-(hydroxymethyl)benzoic acid (C3-3) (5.5 g, 36.1 mmol) in THF (70 mL) at 0 °C were added imidazole (4.92 g, 72.3 mmol) and tert-butylchlorodiphenylsilane (11.92 g, 43.4 mmol). The reaction mixture was stirred at RT for 4 h (monitored by TLC). Then the reaction mixture was dilute with water (100 mL) and extracted with EtOAc (2 x 150 mL). The combined organic layers were washed with saturated NaHCOs solution (2 x 100 mL) dried over anhydrous Na2SO4, filtered and the filtrate was concentrated under reduced pressure. The resulting crude compound was purified by Combi-flash column chromatography (40 g silica column) gradient elution of 35-40 % EtOAc in hexane. The resulting fractions containing desired product were combined and concentrated to afford 3- (hydroxymethyl) benzoic acid (C3-4) (4.0 g, 9.57 mmol, 26.5 % yield) as Off-white solid. LC-MS: Rt = 3.21 m / z 389.2 [M-H]+.Step 4: Synthesis of 3-(methyl(2-(trityloxy)ethyl)amino) propyl 3-(((tert-butyldiphenylsilyl) oxy) methyl) benzoate (C3-6)To a stirred solution of 3-(methyl(2-(trityloxy)ethyl)amino)propan-l-ol (C3-5) (Synthesis described in step-6 to step-8) (1.3 g, 3.46 mmol)) in DCM (20 mL) at 0 °C, were added DCC (2.143 g, 10.39 mmol), DMAP (0.423 g, 3.46 mmol) followed by 3-(((tertbutyldiphenylsilyl)-oxy)methyl)benzoic acid (C3-4) (1.622 g, 4.15 mmol). The reaction mixture was stirred at RT for 16 h (monitored by TLC). Then the reaction mixture was diluted with water (100 mL) and extracted with DCM (2 x 100 mL). The combined organic layers was washed with brine solution (2 x 70 mL) dried over Na2SO4, filtered and the filtrate was concentrated under reduced pressure. The resulting crude compound was purified by Combi-flash column chromatography (12g silica column) gradient elution of 30-40 % EtOAc in hexane. The resulting fractions containing desired product were combined and concentrated to afford 3-(methyl(2- (trityloxy)ethyl)amino)propyl 3-(((tert-butyldiphenylsilyl)-oxy)methyl)benzoate (C3-6) (1.0 g, 1.336 mmol, 38.6 % yield) as Off-white solid. LC-MS: Rt = 2.86 m / z 748.4 [M+H]+.Step 5: Synthesis of 3-(methyl(2-(trityloxy)ethyl)amino)propyl 3-(hydroxymethyl)benzoate (Core-3)To a stirred solution of 3-(methyl(2-(trityloxy)ethyl)amino)propyl 3-(((tert-butyl- diphenylsilyl)oxy)methyl)benzoate (C3-6) (1.0 g, 1.337 mmol) in THF (10 mL) at 0 °C was added TBAF (1.069 mL, 1.069 mmol) and the reaction mixture was stirred at RT for 2 h (monitored by TLC). Then the reaction mixture was quenched with saturated NaHCOs solution (50 mL) and water (30 mL) extracted with EtOAc (2 x 50 mL). The combined organic layers were washed with brine (2 x 70 mL) dried over Na2SO4, filtered and the filtrate was concentrated under reduced pressure. The resulting crude compound was purified by Combi-flash column chromatography (24 g silica column) gradient elution of 50-55 % EtOAc in hexane. The resulting fractions containing desired product were combined and concentrated to afford 3-(methyl (2-(trityloxy)ethyl) amino) propyl 3-(hydroxymethyl) benzoate (Core-3) (0.350 g, 0.661 mmol, 49.5 % yield) as brown semi solid. LC-MS : Rt = 2.21 m / z 510.3 [M+H]+.1H NMR (400 MHz, DMSO-d6) = 6 ppm 7.92 (s, 1H), 7.80-7.78 (d, J = 7.2 Hz, 1H), 7.58-7.57 (d, J = 7.2 Hz, 1H), 7.45 (t, J = 7.8 Hz, 1H), 7.43-7.36 (m, 6H), 7.32-7.29 (m, 6H), 7.25-7.21 (m, 3H), 5.35 (t, J = 5.6 Hz, 1H), 4.56-4.54 (d, J = 6 Hz, 2H), 4.29 (t, J = 6.4 Hz, 2H), 3.03 (t, J =5.6 Hz, 2H), 2.55-2.45 (m, 4 H),2.15 (s, 3H), 1.18-1.16 (m, 2H).Step 6: Synthesis of ethyl 3-((2-hydroxyethyl)(methyl)amino)propanoate (C3-9)A stirred solution of 2-(methylamino)ethan-l-ol (C3-7) (10.0 g, 133.20 mmol) and ethyl acrylate (C3- 8) (15.5 mL, 146.52 mmol) at RT. The reaction mixture was stirred at RT for 30 min (monitored by TLC). Then the reaction mixture was evaporated under reduced pressure and dry under high vacuum to afford ethyl 3-((2-hydroxyethyl)(methyl)amino)propanoate (C3-9) (21.5 g, 122.77 mmol, 92.17% yield) as a colorless liquid.XH NMR (400 MHz, DMSO-d6) = 6 ppm 4.31 (t, J = 5.2 Hz, 1H), 4.04 (q, J = 7.2 Hz, 2H), 3.45-3.40 (m, 2H), 2.60 (t, J = 7.2 Hz, 2H), 2.42-2.36 (m, 4H), 2.15 (s, 3H), 1.17 (t, J = 7.2 Hz, 3H).Step 7: Synthesis of ethyl 3-(methyl(2-(trityloxy)ethyl)amino)propanoate (C3-10)To a stirred solution of ethyl 3-((2-hydroxyethyl)(methyl)amino)propanoate (C3-9) (5.0 g, 28.53 mmol) in pyridine (50 mL) at RT, was added chloromethanetriyl)tribenzene (11.93 g, 42.80 mmol), and the reaction mixture was stirred at 80°C for 16 h (monitored by TLC). Then the reaction mixture was quenched with water (50 mL) extracted with EtOAc (2 x 200 mL). The combined organic layer was washed with saturated NaHCO3 solution (100 mL), followed by brine solution (2 x 100 mL) dried over Na2SO4, filtered and the filtrate was concentrated under reduced pressure. The resulting crudecompound was purified by Combi-flash column chromatography (40 g silica column) gradient elution of 60-70 % EtOAc in hexane. The resulting fractions containing desired product were combined and concentrated to afford ethyl 3-(methyl(2-(trityloxy)ethyl)amino)propanoate (C3-10) (4.0 g, 11.48 mmol, 40.26 % yield) as yellowish oil.TH NMR (400 MHz, DMSO-D6) = 6 ppm 7.40-7.19 (m, 15H), 4.02- 3.96 (m, 2H), 3.02-3.00 (m, 2H), 2.60-2.59 (m, 2H), 2.54-2.53 (m, 2H), 2.41-2.39 (m, 2H), 2.13-2.12 (m, 3H), 1.13-1.09 (m, 3H).Step 8: Synthesis of 3-(methyl(2-(trityloxy)ethyl)amino)propan-l-ol (C3-5)To a stirred solution of ethyl 3-(methyl(2-(trityloxy)ethyl)amino)propanoate (C3-10) (4.0 g, 9.58 mmol) in THF (100.0 mL) at 0 °C, was added LiAIH4(4.79 g, 9.58 mmol). The reaction mixture was stirred at RT for 4 h (monitored by TLC). Then the reaction mixture was quenched slowly with EtOAc (150 mL) washed with water (2 x 100 mL). The organic layer was dried over Na2SO4, filtered and the filtrate was concentrated under reduced pressure. The resulting crude compound was purified by Combi-flash column chromatography (40 g silica column) gradient elution of 25-30% EtOAc in hexane. The resulting fractions containing desired product were combined and concentrated to afford 3-(methyl(2- (trityloxy)ethyl)amino)propan-l-ol (C3-5) (3.2 g, 8.26 mmol, 89.02 % yield) as pale yellow solid. LC-MS: Rt = 2.10 m / z 376.2 [M+H]+.Synthesis of 3-(diethylamino)propyl 4-(hydroxymethyl)benzoate (Core-4)Synthetic Scheme:Step 1: Synthesis of 4-(((tetrahydro-2H-pyran-2-yl)oxy)methyl)benzoic acid (C4-2)To a stirred solution of methyl 4-(hydroxymethyl)benzoic acid (C4-1) (10.0 g, 65.72 mmol) in DCM (100 mL) at 0 °C, were added PPTS (1.6 g, 6.57 mmol) followed by DHP (8.69 mL, 98.58 mmol). The reaction mixture was stirred at RT for 3 h (monitored by TLC). The reaction mixture was diluted with water (150 mL) extracted with DCM (2 x 250 mL). The combined organic layers were washed with saturated NaHCOs solution (2 x 150 mL) dried over anhydrous Na2SO4, filtered and the filtrate was concentrated under vacuum. The resulting crude material was purified by Combi-flash (40 g silica column) with gradient elution of 10% EtOAc in hexane. The resulting fractions containing desired product were combined and concentrated to afford 4-(((tetrahydro-2H-pyran-2-yl)oxy)methyl)benzoic acid (C4-2) (7.0 g, 29.62 mmol, 45.08 % yield) as an off -white solid. MS (LC-MS): m / z 235.0 [M-H]+,TH NMR (400 MHz, DMSO-dg): 6 ppm 12.93 (bs, 1H), 7.92 (d, J = 8.0 Hz, 2H), 7.46 (d, J = 8.4 Hz, 2H), 4.76-4.69 (m, 2H), 4.53 (d, J = 12.8 Hz, 1H), 3.81-3.75 (m, 1H), 3.50-3.45 (m, 1H), 1.76-1.65 (m, 2H), 1.57-1.48 (m, 4H).Step 2: Synthesis of 3-(diethylamino)propyl 4-(((tetrahydro-2H-pyran-2-yl)oxy)methyl) benzoate (C4-4)To a stirred solution of 4-(((tetrahydro-2H-pyran-2-yl)oxy)methyl)benzoic acid (C4-2) (4.99 g, 21.13 mmol) in DCM (50 mL) at 0°C, were added EDC.HCI (6.75 g, 35.22 mmol), DMAP (0.430 g, 3.52 mmol) followed by 3-(diethylamino)propan-l-ol (C4-3) (2.31g, 17.61 mmol). The reaction mixture was stirred at RT for 16 h (monitored by TLC). The reaction mixture was quenched with saturated aqueous NaHCO3solution (100 mL) extracted with DCM (2 x 80 mL). The combined organic layers were dried over sodium sulphate and concentrated under vacuum. The resulting crude material was purified by Combi- flash (40 g silica column) with gradient elution of 70 % EtOAc in hexane. The resulting fractions containing desired product were combined and concentrated to afford 3-(diethylamino)propyl 4- (((tetrahydro-2H-pyran-2-yl)oxy)methyl)benzoate (C4-4) (4.5 g, 12.87 mmol, 73.12% yield) as an off- white solid. MS (LC-MS): m / z 504.3 [M + H]+,TH NMR (400 MHz, DMSO-d6): 6 ppm 7.94 (d, J = 8.4 Hz, 2H), 7.49 (d, J = 8.4 Hz, 2H), 4.77-4.69 (m, 2H), 4.54 (d, J = 13.2 Hz, 1H), 4.29 (t, J = 6.4 Hz, 2H), 3.81- 3.75 (m, 1H), 3.49-3.46 (m, 1H), 2.49-2.41 (m, 6H), 1.82-1.66 (m, 4H), 1.57-1.48 (m, 4H), 0.93 (t, J = 7.2 Hz, 6H).Step 3: Synthesis of 3-(diethylamino)propyl 4-(hydroxymethyl)benzoate (Core 4)To a stirred solution of 3-(diethylamino)propyl 4-(((tetrahydro-2H-pyran-2-yl)oxy)methyl) benzoate (C4-4) (3.6 g, 10.30 mmol) in methanol (150 mL) at 0° C, was added 2M aqueous HCI (15.46 mL, 30.92 mmol). The reaction mixture was stirred at RT for 4 h (monitored by TLC). The reaction mixture was concentrated under vacuum, the resulting residue was diluted with EtOAc (200 mL) washed with saturated aqueous NaHCO3solution (2 x 80 mL). The combined organic layers were dried over sodium sulphate, filtered and the filtrate was concentrated under vacuum 3-(diethylamino)propyl 4- (hydroxymethyl)benzoate (Core 4) (2.3 g, 8.67 mmol, 84.21% yield) as brown oil. MS (LC-MS): m / z 266.15 [M + H]+,XH NMR (400 MHz, DMSO-d6): 6 ppm 7.94 (d, J=8.4Hz, 2H), 7.45 (d, J = 8.0 Hz, 2H), 5.38 (t, J = 5.6 Hz, 1H), 4.57 (d, J = 5.6 Hz, 2H), 4.28 (t, J = 6.4 Hz, 2H), 2.54-2.41 (m, 6H), 1.84-1.77 (m, 2H), 0.93 (t, J = 7.2 Hz, 6H).Synthesis of 4-(diethylamino)butyl 4-(hydroxymethyl)benzoate (Core-5)Synthetic Scheme:Step 1: Synthesis of 4-((tert-butyldiphenylsilyl)methyl)benzoic acid (C5-2)To a stirred solution of 4-(hydroxymethyl)benzoic acid (C5-1) (10.0 g, 65.72 mmol) in DCM (100 mL) at 0°C, were added PPTS (1.6 g, 6.57 mmol) followed by 3,4-dihydro-2H-pyran (8.69 mL, 98.58 mmol).The reaction mixture was stirred at RT for 16 h (monitored by TLC). Then the reaction mixture was diluted with water (100 mL) extracted with DCM (2 x 100 mL). The combined organic layers were washed with saturated NaHCOs solution (2 x 150 mL) dried over Na2SO4, filtered and the filtrate was concentrated under reduced pressure. The resulting crude compound was purified by Combi-flash column chromatography (80 g silica column) gradient elution of 10-10 % EtOAc in hexane. The resulting fractions containing desired product were combined and concentrated to afford 4-((tert- butyldiphenylsilyl)methyl)benzoic acid (C5-2) (7.1 g, 29.620 mmol, 45.72 % yield) as Off -white solid. LC-MS: Rt = 1.61 m / z 235.10 [M-H]+.Step 2: Synthesis of 4-(diethylamino)butyl 4-(((tetrahydro-2H-pyran-2-yl)oxy)methyl)benzoate (C5- 4)To a stirred solution of 4-(((tetrahydro-2H-pyran-2-yl)oxy)methyl)benzoic acid (C5-2) (5.0 g, 21.48 mmol) in DCM (50 mL) at 0 °C, were added EDC.HCI (6.86 g, 35.8 mmol), DMAP (0.437 g, 3.58 mmol) followed by 4-(diethylamino)butan-l-ol (C5-3) ( 2.6 g, 17.90 mmol). The reaction mixture was stirred at RT for 4 h (monitored by TLC). Then the reaction mixture was quenched with saturated NaHCOs solution (150 mL) and was added water (100 mL) then extracted with DCM (2 x 200 mL). The combined organic layers were dried over anhydrous Na2SO4, filtered and the filtrate was concentrated under reduced pressure. The resulting crude compound was purified by Combi-flash column chromatography (80 g silica column) gradient elution of 0 - 7 % Methanol in DCM. The resulting fractions containing desired product were combined and concentrated to afford 4- (diethylamino)butyl 4-(((tetrahydro-2H-pyran-2-yl)oxy)methyl)benzoate (C5-4) (5.1 g, 14.03 mmol, 78.38 % yield) as pale yellow oil. LC-MS: Rt = 1.46 m / z 364.15 [M+H]+.Step 3: Synthesis of 4-(diethylamino)butyl 4-(hydroxymethyl)benzoate (Core-5)To a stirred solution of 4-(diethylamino) butyl 4-(((tetrahydro-2H-pyran-2-yl)oxy) methyl)benzoate (C5-4) (3.0 g, 8.25 mmol) in methanol (50 mL) at 0 °C, was added 2M aqueous HCI (12.37 mL, 24.75 mmol). The reaction mixture was stirred at RT for 4 h (monitored by TLC). Then the reaction mixture was diluted with ethyl acetate (50 mL) and washed with saturated NaHCOs solution (2 x 80 mL). The organic layers was dried over anhydrous Na2SO4, filtered and the filtrate was concentrated under reduced pressure to afford 4-(diethylamino)butyl 4-(hydroxymethyl)benzoate (Core-5) (2.1 g, 7.510 mmol, 91.11 % yield) as pale yellow liquid. LC-MS: Rt = 1.34 m / z 280.15 [M+H]+,TH NMR (400 MHz, CDCI3): 6 ppm 7.93-7.91 (d, J = 8.4 Hz, 2H), 7.47-7.45 (d, J = 8.0 Hz, 2H), 5.39 (s, 1H), 4.62-4.55 (m, 2H), 4.27 (t, J = 6.4 Hz, 2H), 2.48-2.35 (m, 6H), 1.71 (t, J = 6.8 Hz, 2H), 1.49 (t, J = 6.8 Hz, 2H), 0.94-0.91 (m, 6H).Synthesis of 3-(methyl(2-(trityloxy)ethyl)amino)propyl 4-(hydroxymethyl)benzoate (Core-6):Synthetic Scheme:Step 1: Synthesis of ethyl 3-((2-hydroxyethyl)(methyl)amino)propanoate (C6-3)A solution of 2-(methylamino)ethan-l-ol (C6-1) (10.0 g, 133.20 mmol) and acrylate ethyl (C6-2) (15.5 mL, 146.52 mmol) was stirred at RT for 30 min (monitored by TLC). Then the reaction mixture was concentrated under reduced pressure to afford ethyl 3-((2-hydroxyethyl)(methyl)amino)propanoate (C6-3) (21.5 g, 122.77 mmol, 92.17 % yield) as a colorless liquid.TH NMR (400 MHz, DMSO-d6) : 6 ppm 4.31 (t, J = 5.2 Hz, 1H), 4.04 (q, J = 7.2 Hz, 2H), 3.45-3.34 (m, 2H), 2.60 (t, J = 7.2, 2H), 2.44-2.32 (m, 4H), 1.15 (s, 3H), 1.17 (t, J = 7.2, 3H).Step 2: Synthesis of ethyl 3-(methyl(2-(trityloxy)ethyl)amino)propanoate (C6-4)To a stirred solution of ethyl 3-((2-hydroxyethyl)(methyl)amino)propanoate (C6-3) (5.0 g, 28.53 mmol) in pyridine (50.0 mL) at RT, was added chloromethanetriyl)tribenzene (11.93 g, 42.80 mmol) and the reaction mixture was stirred at 80°C for 16 h (monitored by TLC). Then the reaction mixture was quenched with water (50 mL), extracted with EtOAc (2 x 250 mL). The combined organic layers was washed with saturated aq. NaHCOs solution (100 mL) followed by saturated brine solution (100 mL), dried over Na2SO4, filtered and filtrate was concentrated under reduced pressure afford ethyl 3- (methyl(2-(trityloxy)ethyl)amino)propanoate (C6-4) (4.5 g, 12.92 mmol, 45.30 % yield) as yellowish oil.XH NMR (400 MHz, DMSO-d6): 6 ppm 7.45-7.37 (m, 6H), 7.35-7.29 (m, 6H), 7.27-7.24 (m, 3H), 4.01- 3.96 (q, J = 7.2 Hz, 2H), 3.00 (t, J = 5.6 Hz, 2H), 2.66-2.52 (m, 4H), 2.44-2.37 (m, 2H), 2.12 (s, 3H), 1.11 (t, J = 7.2, 3H).Step 3: Synthesis of 3-(methyl(2-(trityloxy)ethyl)amino)propan-l-ol (C6-5)To a stirred solution of ethyl 3-(methyl(2-(trityloxy)ethyl)amino)propanoate (C6-4) (4.0 g, 9.58 mmol) in THF (100 mL) at 0 °C, was added LAH (4.79 g, 9.58 mmol) and the reaction mixture was stirred at RT for 4 h (monitored by TLC). Then the reaction mixture was quenched with EtOAc (150 mL), washed with water (2 x 150 mL). The combined organic layers were dried over NajSO^ filtered and filtrate was concentrated under reduced pressure. The resulting residue was purified by Combi-flash column chromatography (40 g silica column) with gradient elution of 1-10% EtOAc in hexane. The resulting fractions containing desired product were combined and concentrated to afford 3-(methyl(2- (trityloxy)ethyl)amino)propan-l-ol (C6-5) (3.1 g, 8.26 mmol, 86.24% yield) as pale yellow solid. MS (LCMS): m / z 376.25 [M-H]+.Step 4: Synthesis of 4-(((tert-butyldiphenylsilyl)oxy)methyl)benzoic acid (C6-7)To a stirred solution of 4-(hydroxymethyl)benzoic acid (C6-6) (1.0 g, 6.57 mmol) in THF (20.0 mL) at 0°C , was added imidazole (0.894 g, 13.14 mmol) followed by TBDPS-CI (1.9 g, 7.22 mmol) and the reaction mixture was stirred at RT for 3 h (monitored byTLC). Then the reaction mixture was quenched with water (50 ml), extract with EtOAc (2 x 50 mL). The combined organic layer was washed with saturated aq. NaHCOs solution (50 mL), Then, the organic layers were dried over NajSC , filtered and filtrate was concentrated under reduced pressure. The resulting residue was purified by Combi-flash column chromatography (40 g silica column) with gradient elution of 1-7 % EtOAc in hexane. The resulting fractions containing desired product were combined and concentrated to afford 4-(((tert- butyldiphenylsilyl)oxy)methyl)benzoic acid (C6-7) (0.450 g, 1.15 mmol, 17.53 % yield) as an Off-white solid. MS (LCMS): m / z 389.2 [M-H]+.Step 5: Synthesis of 3-(methyl (4,4,4-triphenylbutyl)amino)propyl 4 (((tertbutyldiphenylsilyl) oxy)methyl) benzoate (C6-8)To a stirred solution of 4-(((tert-butyldiphenylsilyl)oxy)methyl)benzoic acid (C6-7) (0.3 g, 0.79 mmol) in DCM (10 mL) at 0 °C, was added EDC.HCI (0.302 g,1.58 mmol) , DMAP (0.019 g, 0.150 mmol, 0.2) followed by 3- (methyl(2-(trityloxy)ethyl)amino)propan-l-ol (C6-5) ( 0.374 g, 0.95 mmol) and the reaction mixture was stirred at RT for 16 h (monitored by TLC). Then the reaction mixture was quenched with saturated NaHCOs solution (50 mL), was added water (20 mL) and extracted with DCM (2 x 50 mL). The combined organic layer was dried over Na2SO4, filtered and filtrate was concentrated under reduced pressure. The resulting residue was purified by Combi-flash column chromatography (40 g silica column) with gradient elution of 1-40 % EtOAc in hexane. The resulting fractions containing desired product were combined and concentrated to afford 3-(methyl(4,4,4-triphenylbutyl) amino) propyl 4-(((tert-butyldiphenylsilyl)oxy)methyl)benzoate (C6-8) (0.310 g, 0.414 mmol, 52.45 % yield) as pale yellow semi solid. MS (LCMS): m / z 748.40 [M+H]+.Step 6: Synthesis of 3-(methyl(2-(trityloxy)ethyl)amino)propyl 4-(hydroxymethyl)benzoate (Core-6)To a stirred solution of 3-(methyl(2-(trityloxy)ethyl)amino)propyl 4-(((tert butyldiphenylsilyl)oxy) methyl) benzoate (C6-8) (0.25 g, 0.334 mmol) in THF (10 mL) at 0 °C ,was added TBAF (0.26 mL, 0.267 mmol) and the reaction mixture was stirred at RT for 2 h (monitored by TLC). Then the reaction mixture was quenched with saturated aq. NaHCO3 solution (50 mL), was added water (20 mL), extracted with EtOAc (2 x 50 mL). The combined organic layers were dried over NajSO^ filtered and filtrate was concentrated under reduced pressure. The resulting residue was purified by Combi-flash column chromatography (40 g silica column) with gradient elution of 1 - 55 % EtOAc in hexane. The resulting fractions containing desired product were combined and concentrated to afford 3-(methyl(2- (trityloxy)ethyl)amino)propyl 4-(hydroxymethyl)benzoate (Core-6) (0.1 g, 0.414 mmol, 59 % yield) as Brown semi solid. MS (LCMS): m / z 510.35 [M+H]+,TH NMR (400 MHz, CDCI3) : 6 ppm 8.01-7.99 (d, J = 7.2 Hz, 2H), 7.48-7.36 (m, 9H), 7.34-7.27 (m, 6H), 7.7-7.16 (m, 3H), 4.77 (s, 2H), 4.34 (t, J = 6.8 Hz, 2H), 3.20 (t, J = 6 Hz, 2H), 2.64 (t, J = 6.4 Hz, 2H), 2.52 (t, J = 7.2 Hz, 2H),2.24 (s, 1H), 1.91 (t, J = 7.6 Hz, 2H), 0.88 (t, J = 6.4 Hz, 3H).Synthesis of 3-(diethylamino)propyl 3-(6-((((9Z,12Z)-octadeca-9,12-dienoyl)oxy)methyl)-12- (octyloxy)-3,9-dioxo-2,4,8,13-tetraoxahenicosyl)benzoate (Compound 2-10)Step 1: Synthesis of 3-(diethylamino)propyl 3-(6-((((9Z,12Z)-octadeca-9,12-dienoyl)oxy)-methyl)-12- (octyloxy)-3,9-dioxo-2,4,8,13-tetraoxahenicosyl)benzoate (Compound 2-10)To a stirred solution of 3-((4,4-bis(octyloxy)butanoyl)oxy)-2-(hydroxymethyl)propyl (9Z,12Z)- octadeca-9,12-dienoate (Lipid B) (500 mg, 0.719 mmol) in DCM (10 ml) at 0 °C were added Pyridine (0.059 ml, 0.719 mmol) and DMAP (88 mg, 0.719 mmol) followed by 4-Nitrophenyl chloroformate (145 mg, 0.719 mmol). The reaction mixture was stirred at RT for 2 h, then were added DIPEA (0.123 ml, 0.719 mmol) and 3-(diethylamino)propyl 3-(hydroxymethyl)benzoate (Core 1) (Synthesis described separately for Core 1) (191 mg, 0.719 mmol) then stirring continued for further 14 h (monitored by TLC). The Reaction mixture was diluted with water (100 mL) extracted with DCM (2 X 100 mL). The combined organic layer was washed with brine solution (50 mL). The organic layer was dried over anhydrous NajSCU, filtered and the filtrate was concentrated under vacuum. The resulting crude material was purified by Combi-flash (24 g silica column) with gradient elution of 2% MeOH in DCM. The resulting fractions containing desired product were combined and concentrated to afford 3- (diethylamino)propyl 3-(6-((((9Z,12Z)-octadeca-9,12-dienoyl)oxy)methyl)-12-(octyloxy)-3,9-dioxo- 2,4,8, 13-tetraoxahenicosyl) benzoate (220 mg, 0.221 mmol, 30.8 % yield) as pale yellow liquid. MS (ESI): m / z 987.15 [M+H]+, HPLC-CAD purity: 98.22%, HPLC-ELSD purity: 99.25%,TH NMR (400 MHz, CDCI3) = 6 ppm 8.04- 8.00 (m, 2H), 7.61-7.55 (m, 1H), 7.48-7.46 (m, 2H), 5.37-5.32 (m, 4H), 5.19 (s, 2H), 4.47 (t, J = 5.6 Hz, 1H), 4.38-4.36 (m, 2H), 4.25-4.10 (m, 2H), 4.14-4.13 (m, 4H), 3.56-3.52 (m, 2H), 3.42- 3.36 (m, 2H), 2.78-2.75 (m, 2H), 2.61-2.58 (m, 6H), 2.41-2.39 (m, 3H), 2.29-2.27 (m, 2H), 2.05-2.02 (m, 4H), 1.94-1.91 (m, 4H), 1.58-1.53 (m, 6H), 1.33-1.26 (m, 33H), 1.06 (t, J = 7.2 Hz, 6H), 0.90-0.85 (m, 9H).Synthesis of ((((3-((3-(diethylamino)propoxy)carbonyl)benzyl)oxy)carbonyl)azanediyl) bis(hexane-6,l-diyl) bis(Z-hexyldecanoate) (Compound 2-11)Step 1: Synthesis of ((((3-((3-(diethylamino)propoxy)carbonyl)benzyl)oxy)carbonyl)azanediyl) bis(hexane-6,l-diyl) bis(2-hexyldecanoate) (Compound 2-11)To a stirred solution of 3 3-(diethylamino)propyl 3-(hydroxymethyl)benzoate (Core-1) (Synthesis described separately for Core-1) (300 mg, 1.130 mmol. 1.0 eq.) in DCM (10 mL) at 0 °C were added pyridine (0.183 ml, 2.261 mmol, 2.0 eq.) and DMAP (41 mg, 0.339 mmol, 0.3 eq.) followed by 4- Nitrophenyl chloroformate (273 mg, 1.356 mmol, 1.2 eq.). The reaction mixture was stirred at RT for 16 h, then were added DIPEA (0.394 ml, 2.261mmol, 2.0 eq.) and a solution of ((((3-((3- (diethylamino)propoxy)carbonyl)benzyl)-oxy)carbonyl)azanediyl)bis(hexane-6,l-diyl) bis(2- hexyldecanoate) (Lipid C) (550 mg, 0.791 mmol, 0.7 eq.) in DCM (5 mL) then stirring continued for further 16 h (monitored by TLC). The reaction mixture was diluted with DCM (200 mL) washed with water (3 x 75 mL). The organic layer was dried over anhydrous NajSC , filtered and the filtrate was concentrated under vacuum. The resulting crude material was purified by Combi-flash (24 g silica column) with gradient elution of 4-7% MeOH in DCM. The resulting fractions containing desired product were combined and concentrated to afford ((((3-((3- (diethylamino)propoxy)carbonyl)benzyl)oxy)carbonyl)azanediyl)bis(hexane-6,l-diyl) bis(2- hexyldecanoate) (Compound 2-11) (0.160 g, 0.162 mmol, 14% yield) as yellow syrup, which was confirmed byXH NMR. HPLC-ELSD purity: 98.24%.TH NMR (400 MHz, CDCI3) = 6 ppm 8.02-8.00 (m, 1H), 7.97-7.95 (m, 1H), 7.55-7.51 (m, 1H), 7.45-7.41 (m, 1H), 5.15 (s, 2H), 4.38 (t, J = 6.4 Hz, 2H), 4.08- 4.02 (m, 4H), 3.60-3.40 (m, 2H), 3.28-3.18 (m, 4H), 2.70-2.50 (m, 6H), 2.32-2.29 (m, 2H), 1.62-1.58 (m, 8H), 1.54-1.49 (m, 2H), 1.44-1.37 (m, 10H), 1.30-1.20 (m, 44H), 1.15-1.00 (m, 6H), 0.88-0.85 (m, 12H).Synthesis of ((((3-((4-(diethylamino)butoxy)carbonyl)benzyl)oxy)carbonyl)azanediyl)bis-(hexane- 6,1-diyl) bis(2-hexyldecanoate) (Compound 2-12)Compound 2-12Synthetic Scheme:Step 1: Synthesis of ((((3-((4-(diethylamino)butoxy)carbonyl)benzyl)oxy)carbonyl)azanediyl) bis(hexane-6,l-diyl)bis(2-hexyldecanoate) (Compound 2-12)To a stirred solution of 4-(diethylamino)butyl 3-(hydroxymethyl)benzoate (Core-2) (Synthesis described separately for Core-2) (0.300 g, 1.074 mmol) in DCM (10 mL) at 0 °C, were added Pyridine (0.346 mL, 4.30 mmol), DMAP (0.039 g, 0.322 mmol) followed by 4-Nitrophenyl chloroformate (0.325 g, 1.611 mmol) and the reaction was warmed to stir at RT for 16 h (monitored by TLC). Then the reaction mixture was added DIPEA (0.750 mL, 4.30 mmol) followed by azanediylbis(hexane-6,l-diyl) bis(2-hexyldecanoate) (Lipid C) (0.596 g, 0.859 mmol) and stirred at RT for another 24 hr (monitored by TLC). The reaction mixture was diluted with water (50 mL) and extracted with DCM (2 x 50 mL). The combined organic layer was washed with brine solution, dried over anhydrous NajSCU, filtered and the filtrate was concentrated under vacuum. The resulting crude was material was purified by Combi- flash (silica column) with gradient elution of EtOAc in hexane followed by 0-5 % MeOH in DCM. The resulting fractions containing desired product were combined and concentrated to afford ((((3-((4- (diethylamino)butoxy)carbonyl)- benzyl)oxy)-carbonyl)azanediyl)Bis(hexane-6,l-diyl)bis(2- hexyldecanoate) (Compound 2-12): (0.100 g, 0.099 mmol, 9.20% yield) as pale yellow color syrup. MS (ESI): m / z 1000.15 [M+H]+. HPLC-ELSD purity: 98.73%.TH NMR (400 MHz, CDCI3) = 6 ppm 8.00-7.96 (m, 2H), 7.56-7.52 (m, 1 H), 7.45-7.41 (m, 1H), 5.16 (s, 2H), 4.35 (t, J = 5.6 Hz, 2H), 4.05 (m, 4H), 3.26- 3.22 (m, 4H), 3.05-2.55 (m, 6H), 2.31-2.27 (m, 2H), 1.89-1.70 (m, 4H), 1.72-1.58 (m, 12H), 1.52-1.16 (m, 58 H), 0.88-0.85 (m, 12H).Synthesis of ((((3-((3-((2-hydroxyethyl) (methyl)amino) propoxy)carbonyl)benzyl)oxy)carbonyl) azanediyl) bis (hexane-6,l-diyl) bis (2-hexyldecanoate) (Compound 2-13)Compound 2-13Synthetic Scheme:Step 1: Synthesis of ((((3-((3-(methyl(2-(trityloxy)ethyl)amino) propoxy) carbonyl)benzyl) oxy)carbonyl)azanediyl)bis(hexane-6,l-diyl) bis(2-hexyldecanoate) (23-2)To a stirred solution of 3-(methyl(2-(trityloxy)ethyl)amino)propyl 3-(hydroxymethyl-)benzoate (Core- 3, 23-1) (0.300 g, 0.589 mmol) in DCM (20 mL) at 0 °C, was added pyridine (0.095 mL, 1.177 mmol), DMAP (0.022 g, 0.177 mmol) followed 4- Nitrophenyl chloroformate (0.178 g, 0.883 mmol) and the reaction mixture was stirred at RT for 2 h (monitored by TLC). Then to the above reaction mixture was added a solution of DIPEA (0.206 mL, 1.177 mmol), azanediylbis(hexane-6,l-diyl) bis(2- hexyldecanoate) (Lipid-C) (0.327 g, 0.471 mmol) and DCM (5 mL). The reaction mixture was stirred at RT for 16 h (monitored by TLC). Then the reaction mixture was diluted with water (40 mL), extracted with DCM (2 x 40 mL). The combined organic layer was washed with brine solution (50 mL), dried over Na2SO4, filtered and the filtrate was concentrated under reduced pressure. The resulting crude compound was purified by Combi-flash column chromatography (24 g silica column) gradient elution of 5-7 % MeOH in DCM. The resulting fractions containing desired product were combined and concentrated to afford ((((3-((3-(methyl(2-(trityloxy)ethyl)amino) propoxy) carbonyl)benzyl) oxy)carbonyl)azanediyl)bis(hexane-6,l-diyl) bis(2-hexyldecanoate) (23-2) (0.210 g, 0.017 mmol, 29.0 % yield) as colorless syrup.TH NMR (400 MHz, CDCI3): 6 ppm 7.99 (s, 1H), 7.95-7.93 (d, J = 8 Hz, 1H), 7.55-7.53 (d, J = 8 Hz, 1H), 7.48-7.38 (m, 7H), 7.32-7.28 (m, 6 H), 7.24-7.18 (m, 3H), 5.15 (s, 2H), 4.34 (t, J = 6.8 Hz, 2H), 4.10-3.98 (m, 4H), 3.32-3.18 (m, 6 H), 2.64 (t, J = 6.4 Hz, 2H), 2.51 (t, J = 6.8 Hz, 2H), 2.29-2.27 (m, 2H), 2.23 (s, 3H), 1.92-1.89 (m, 2H), 1.68-1.58 (m, 8 H), 1.57-1.54 (m, 3H), 1.48-1.38 (m, 6H), 1.37-1.15 (m, 48H), 0.88-0.85 (m, 12H).Step 2: Synthesis of ((((3-((3-((2-hydroxyethyl) (methyl)amino) propoxy)carbonyl)benzyl)...

Claims

1. CLAIMSWe claim:

1. An (ionizable) cationic lipid compound represented by a structural formula selected from any one of:wherein,R1 is selected from the group consisting of:R20 is R19 or R24;R21 is selected from: a bond, -CH2-, -[CHzh-, -C(=O)-, -S-, -S(=O)- or -S(=O)2-;R22 is selected from: a bond (no atoms in between), optionally substituted Ci-Cg alkylene, C1-C10 alkyleneoxy, -C(=O)-, -R143-C0-C3alkylene-R146-C0-C3alkylene-, -CH(OH)-, -C(OH)2-, -C(R25)2-, - CH(OR25)-, -C(OR25)2-, -N(R25)-, -N(R116)C(=O)-, -C(=O)N(R116)-, -O-, -S-, -S(=O)-, -S(=O)2-, - P(OR25)- or -P(O)(OR25)-;R116 is -H or Ci.3alkyl;R143 is selected from: a bond, -O-, -CH(OH)-, -C(OH)2-, -C(=O)-, -C(=O)O-, -OC(=O)-, -OC(=O)O-, - N(R116)-, -N(R116)C(=O)-, -C(=O)N(R116)-, -S-, -S-S-, -C(=S)-, -C(=O)-C-,-CH(OH)-, -S-C(=O)-, or - C(=O)-S-;R146 is an optionally substituted group selected from: 3-7 membered carbocyclylene or 3-7 membered heterocyclylene;R25, for each occurrence, is independently selected from: a bond, -H, -CD3, -CECH, -CH2CH3, -F, -Cl, - Br, -I, -OCH3, -C(R114)3, -NHC(=O)R114, -OR114, -N(R114)2, -SR114, -S(=O)R114, -S(=O)2R114, - P(R114)2, -P(OR114)2, -P(O)(R114)2, -Si(R114)3, -Si(O)2R114, or an optionally substituted group selected from: linear or branched C1-C10- alkyl, linear or branched C2-C10 alkenyl, linear or branched C2-C10 alkynyl, a 3-18-membered saturated or partially unsaturated carbocyclyl or heterocyclyl, or a 5-18-membered unsaturated carbocyclyl or heterocyclyl; wherein said heterocyclyl contains one or more heteroatoms independently selected from N, O or S as a ring atom;R114, for each occurrence, is independently selected from: -H, -CH2(halo), -CH(halo)2, -C(halo)3, -F, - CF3, -Cl, -Br, -I, -OH, or an optionally substituted, linear or branched, group selected from C1-C6 alkyl, C2-C6alkenyl or C2-C6alkynyl;R23 is an optionally substituted group selected from: a 3-18-membered saturated or partially unsaturated carbocyclyl, a 5-18-membered unsaturated carbocyclyl, a 3-18-membered saturated or partially unsaturated heterocyclyl, or 5-18-membered unsaturated heterocyclyl; wherein said heterocyclyl contains one or more of heteroatoms independently selected from N, O or 8 as a ring atom;R24 for each occurrence, is independently selected from: a bond, -H, -CH2(halo), -CH(halo)2, - C(halo)3, -F, -CF3, -Cl, -Br, -I, -C(R25)3, -OR25, -N(R25)2, -SR25, -S(=O)R25, -S(=O)2R25, - OS(=O)2NHR25, -S(=O)2NHR25, -P(R25)2, -P(R25)2, -P(OR25)2, -P(O)(R25)2, -P(O)(OR25)2, - Si(R25)3or an optionally substituted group selected from: linear or branched C1-C10- alkyl, linear or branched C2-Ci0alkenyl, linear or branched C2-Ci0alkynyl;R2 is a bond, or a linear, branched or optionally substituted group selected from: -C1-C15 acyclic hydrocarbylene-, Ci-Cg alkylene, C2-C8 alkenylene, C2-C8 alkynylene, -Co-Cg acyclic hydrocarbylene-R148-C0-Cg acyclic hydrocarbylene-, 3-7 membered carbocyclylene or 3-7 membered heterocyclylene;R148 is selected from: R47, R143, R146, -CH(R147)-, -O-, -S-, -S-S- or -N(R114)-;R47, for each occurrence, is independently selected from: a bond, an optionally substituted 3-14- membered saturated, partially unsaturated or unsaturated carbocyclene or an optionally substituted 3-14-membered saturated, partially unsaturated or unsaturated heterocyclene;R147 is an optionally substituted group selected from: 3-7 membered carbocyclyl or 3-7 membered heterocyclyl; each R50 is independently selected from:-R62-R123-R63-R66,-R62-R123-R63-R74-R66,-R62-R123-R63-R124-R64-R65-R66,-R62-R123-R63-R124-R64-R74-R66,-R62-R123-R63-R124-R64-R74-R127-R66,-R62-R123-R63-R124-R64-R74-R127-R75-R66, or-R62-R123-R63-R124-R64-R74-R127-R75-R65-R66; wherein,R3, R4, Rll and R12 are each independently selected from: a bond, R26-Ci-Cio alkylene-R27, R26-C2- Cio alkenylene-R27, R26-C2-CIO alkynylene-R27, R26-C3-C10 carbocyclylene-R27, R26-C3-C10 heterocyclylene-R27R26-CH=CH-CH2-R47-R27, R26-R47-CH2-CH=CH-R27, R26-R47-CH2-R47- R27, R26-CH=CH-CD2-R47-R27, R26-R47-CD2-CH=CH-R27 or R26-R47-CD2-R47-R27; wherein, R47 for each occurrence, is an optionally substituted group independently selected from 3-10- membered carbocyclylene or 3-10-membered heterocyclylene;R26 and R27 are each independently selected from: a bond, or an optionally substituted, linear or branched, C1-C10 alkylene, C1-C10 alkenylene or C1-C10 alkynylene;R5, R6, R9 and RIO are each independently selected from: a bond, -CH2-, -O-, -S-, -CH(OH)-, - CH(NH2)-, -NH-C(=O)-O-, -O-C(=O)-NH-, -C(NH[R25])-, -C(NH[OR25])-, -C(=O)-, -O-C(R25)2-, -O- C(=O)-, -C(=O)-O-, -C(=S)-, -N(R25)-, -S-S-, -S-CH(CH3)2-S-; -C(F)2-C(=O)-O- or -O-C(=O)-C(F)2-;R7 and R8 are each independently selected from: a bond, R3, -CH2-, -C(=O)-, -C(=S)- -CH(R25)-, - C(R25)2-, -CH(OR25)- , -CH(NH[R25])-, -C(NH[OR25])-, -N(R25)-, -O-, -S-, -S(OH)-, -S(=O)-, - S(=O)2-, -S(OH)2-, -C(=O)C(R25)2C(=O)-, -C(R25)2C(=O)-, -C(=O)C(R25)2-O-, -C(R25)2C(R25)2-, - C(R25)2-O-, -O-C(R25)2C(R25)2-, -O-C(R25)2C(=O)-, -C(=O)-O-R115-O-C(=O)-, -P(OH)-, - P(OH)(R25)-, -P(=O)(OH)-, -P(OR25)2-, -P(OR25)-, -P(=O)(OR25)-, -C(F)2C(=O)-O- or -O- C(=O)C(F)2-;R115, for each occurrence, are each independently selected from: R26-Ci-Cg alkylene-R27, R26-C2-C8 alkenylene-R27, R26-C2-C8alkynylene-R27, R26-R47-R27, R26-CH=CH-CH2-R47-R27, R26-R47- CH2-CH=CH-R27, R26-R47-CH2-R47-R27, R26-CH=CH-CD2-R47-R27, R26-R47-CD2-CH=CH-R27 or R26-R47-CD2-R47-R27; wherein, R47 for each occurrence, is an optionally substituted group independently selected from 3-10-membered carbocyclylene or 3-10-membered heterocyclylene; and each double bond or cPr has a cis configuration;R13, R14, R61, R73 and R107 are each independently selected from:R60 is selected from: is absent (i.e., an unbonded pair of electrons on N), -H or optionally substituted C1-C6 alkyl; wherein, when R60 is -H or optionally substituted C1-C6 alkyl, R60 is bound directly to a nitrogen atom with a +1 charge, wherein the +1 charge may be counterbalanced with anion from a pharmaceutically acceptable acid;R15, R16, R17 and R18 are each independently selected from: -H; an optionally substituted linear or branched, -R26-Ci-Ci0alkylene-R28, -R26-C2-CI0alkenylene-R28, or -R26-C2-CI0alkynylene- R28; an optionally substituted, R26-R149, R26-R23, -R26-C3-C7 cycloalkylene-R28, -R26-C3-C7 cycloalkylene-R26-C3-C7 cycloalkylene-R28 -R26-C3-C7 cycloalkenylene-R28, -R26-C5-Ci5- spirocycloalkylene-R28, -R26-C3-Ci5-carbocyclylene-R28, -R26-C3-Ci5-heterocyclylene-R28, - R26-arylene-R28, or -R26-heteroarylene-R28; or -R26-CH=CH-CH2-cPr-R28, -R26-cPr-CH2- CH=CH-R28, -R26-cPr-CH2-cPr-R28, -R26-CH=CH-CD2-R47-R28, -R26-R47-R47-R28, -R26-R47- R28, -R26-R47-R26-R47-R28, -R26-CD2-R47-R28, -R26-R47-CD2-R47-R28, -R26-R47-CD2-CH=CH- R28, -R26-CH=CH-CH2-CH=CH-R28, -R26-CH=CH-CH2-R47-R28, -R26-R47-CH2-CH=CH-R28, -R26- R47-CH2-R47-R28, -R26-CH=CH-CD2-CH=CH-R28;R28 is selected from: -H, C1-C10 alkyl, C2-C 10 alkenyl or C2-Cio alkynyl;R149 is:each R155 is each independently selected from: -H, -OH, optionally substituted C1-C6 alkyl, optionally substituted Cj-Cg alkenyl, optionally substituted Cj-Cg alkynyl or optionally substituted C1-C6 alkoxy;R150 is selected from optionally substituted C1-C15 alkyl, optionally substituted C2-Ci5alkenyl, optionally substituted C2-C15 alkynyl, optionally substituted C1-C15 alkoxy, optionally substituted C1-C10 alkylene-C(O)O-optionally substituted C1-C15 alkyl or optionally substituted C1-C10 alkylene-COOH;R151 is -H and R152 is -OH, or R151 and R152 are both -H, or R151 and R152 are both absent; when R151 and R152 are absent a C=C double bond is present between the carbon atoms that R151 and R152 are bound to; and,R153 is -OH and R154 is -H, or R153 and R154 are both -H, or R153 and R154 are both absent, wherein when R153 and R154 are absent then a C=C double bond, preferably cis (Z configuration stereochemistry) is present between the carbon atoms to which R153 and R154 are bound;R62, R63, R64, R67, R68, R69, R74, R75, R76, R77, R108, R109, R110 and Rill are each independently selected from: a bond, C1-C10 alkylene, C2-C10 alkenylene, C2-C10 alkynylene, C3- C7cycloalkylene, C3-C7 cycloalkenylene, C5-Cio-spirocycloalkylene, Cs-Cw-carbocyclylene, C3- Cio-heterocyclylene, -R26-CH=CH-C2-C6alkylene-CH=CH-R27-, -R26-CH=CH-C2-C6alkylene-R47- R27-, -R26-R47-C2-C6alkylene-CH=CH-R27-, -R26-R47-C2-C6alkylene-R47-R27-, -R26-C3-C6- cycloakylene-R27-, -R26-CH=CH-CH2-CH=CH-R27-, -R26-CH=CH-CH2-R47-R27-, -R26-R47-CH2- CH=CH-R27-, -R26-R47-CH2-R47-R27-, -R26-CH=CH-CD2-CH=CH-R27-, -R26-CH=CH-CD2-R47- R27-, -R26-R47-CD2-CH=CH-R27- or -R26-R47-CD2-R47-R27-; wherein, R47 for each occurrence, is an optionally substituted group independently selected from 3-10-membered carbocyclylene or 3-10-membered heterocyclylene; each double bond or cPr has a cis configuration;R65, R70, R78, R105 and R112 are each independently selected from: a bond, C1-C10 alkylene, C2-C10 alkenylene, C2-C10 alkynylene, C3-C8cycloalkylene, C3-C8cycloalkenylene, C5-C10- spirocycloalkylene, Cs-Cw-carbocyclylene, Cs-Cio-heterocyclylene, R26-CH=CH-C2-Cgalkylene- CH=CH-R27-, -R26-CH=CH-C2-C6alkylene-R47-R27-, -R26-R47-C2-C6alkylene-CH=CH-R27-, -R26- R47-C2-C6alkylene-R47-R27-, -CH=CH-CH2-CH=CH-, -CH=CH-CH2-R47-, -R47-CH2-CH=CH-, -R47- CH2-R47-, -CH=CH-CD2-CH=CH- , -CH=CH-CD2-R47-, -R47-CD2-CH=CH- or -R47-CD2-R47-, wherein, R26 and R27 are each independently selected from: a bond, C1-C10 alkylene, C2-C10 alkenylene or C2-C10 alkynylene; R47 for each occurrence, is an optionally substituted groupindependently selected from 3-10-membered carbocyclylene or 3-10-membered heterocyclylene;R66, R71, R79, R106 and R113 are each independently selected from: -H, linear or branched C1-C10 alkyl, C2-Cio alkenyl, or C2-Ci0alkynyl; or C5-Ci5-spirocycloalkylene-R28, C3-Ci5-carbocyclylene- R28, C3-Ci5-heterocyclylene-R28, -arylene-R28, or -heteroarylene-R28, -R26-R23, -R26-CH=CH- CH2-cPr-R28, -R26-cPr-CH2-CH=CH-R28, -R26-cPr-CH2-cPr-R28, R26-R149, -R26-R23, -R26- CH=CH-CH2-CH=CH-R28, -R26-CH=CH-CH2-R47-R28, -R26-R47-CH2-CH=CH-R28, -R26-R47-CH2- R47-R28, -R26-CH=CH-CD2-CH=CH-R28, -R26-CH=CH-CD2-R47-R28, -R26-R47-R47-R28, -R26- R47-R26-R47-R28, -R26-CD2-R47-R28, -R26-R47-CD2-R47-R28, -R26-R47-CD2-CH=CH-R28 or - R26-C3-C6cycloakylene-R26-C3-C6cycloakylene-R28;R123, R124, R125, R126, R127, R128, R132 and R133 are each independently is selected from: a bond, -O-, -OC(=O)-, -C(=O)O-, -OC(=O)O-, -S-C(=O)-, -C(=O)-S-, -S-S-, -C(=O)N(R164)-, - N(R164)C(=O)-, -N(R164)C(=O)N(R164)-, -O-C(=O)C(R164)2C(=O)O-, -C(=O)O-C(R164)2C(=O)O-, - O-C(=O)C(R164)2-O-C(=O)-, -O-C(R164)2C(=O)O-, -O-C(=O)C(R164)2C-O-, -O-C(R164)2-O-C(=O)-, -C(=O)-O-C(R164)2C-O-, -O-C(=O)-O-R191-O-C(=O)O-, -C(=O)-, -OC(=S)O-, -C(=S)O-,-OC(=S)- - C(=S)- -CH(R25)-, -C(R25)2-, -CH(OR25)-, -CH(NH[R25])-, -C(NH[OR25])-, -N(R25)-, -S-, -S(=O)-, - S(=O)2-, -OS(=O)2NH2-, -S(=O)2NH2, -S(OH)-, -S(OH)2-, -C(=O)C(R25)2C(=O)-, -C(R25)2C(=O)-, - C(=O)C(R25)2-O-, -C(R25)2C(R25)2-, -C(R25)2-O-, -O-C(R25)2C(R25)2-, -O-C(R25)2C(=O)-, -C(=O)- O-R115-O-C(=O)-, -P(OH)(R25)-, -P(OR25)2-, -P(OR25)- and -P(=O)(OR25)-, -C(CI)2-C(=O)-O-, -O- C(=O)-C(CI)2-, -C(F)2-C(=O)-O-, -O-C(=O)-C(F)2-;R164, for each occurrence, is independently selected from: -H or optionally substituted, linear or branched C1-C6 alkyl;R191 is selected from: is a bond or optionally substituted, linear or branched C1-C6 alkylene, -C0-C3- alkylene-C3-Cio-carbocyclylene-Co-C3-alkylene- or -Co-C3-alkylene-C3-Cio-heterocyclylene-Co-C3- alkylene-; each R137 is independently selected from:R142 is selected from: -N(R31)(R32), R58, R119 or R32;R31 and R32 are each independently selected from: -H or an optionally substituted group selected from: linear or branched Ci-io alkyl, Ci-io hydroxyalkyl, Ci-io alkoxyalkyl, linear or branched Ci-io alkenyl, linear or branched Ci-io alkynyl, 3-14 membered carbocyclyl, 3-14 membered heterocyclyl, 5-10 membered spirocyclyl, 5-10 membered spiroheterocyclyl, 3-7 memberedcarbocyclyl-Ci.6alkyl-, 3-7 membered heterocyclyl-Ci.6alkyl-, 5-10 membered spirocyclyl-Ci.6alkyl-, 5-10 membered spiroheterocyclyl-Ci.g alkyl-, or, R31 and R32, together with the nitrogen atom to which they are attached, form a 3-14 membered heterocyclyl, 5-10 membered spiroheterocyclyl, 3-7 membered heterocyclyl-Ci.6alkyl- or 5-10 membered spiroheterocyclyl-Ci-6 alkyl-;R34 is selected from a bond, -N(R116)-, -O-, -S-, -S(R46);R116 is -H or C1.3 alkyl;R46 is selected from: absent (i.e., two lone electron pairs), =0 or (=0)2;R120 is selected from: a bond, -C(=O)O-, -OC(=O)-, -O-CH2-, -O-, -N(R116)C(=O)-, -C(=O)N(R116)-, - C(=O)O-, -S-S-, -C(=O)- or -C(=O)-CH2-;R47 is selected from: a bond, an optionally substituted 3-14-membered saturated, partially unsaturated or unsaturated carbocyclene or an optionally substituted 3-14-membered saturated, partially unsaturated or unsaturated heterocyclene;R48 is an optionally substituted group selected from: a bond, C1-10 alkylene, C1-10 internal alkenylene, Ci-10 internal alkynylene, -C0-C3 alkyl-3-6-membered cycloalkylene-Co-Cs alkyl-, -C0-C3 alkyl-4-6- membered cycloalkenylene- C0-C3alkyl-, -C0-C3alkyl-phenylene- C0-C3alkyl-, -C0-C3alkyl-3-6- membered heterocycloalkylene-Co-Cs alkyl-, -C0-C3 alkyl-4-6-membered heterocycloalkenylene- C0-C3 alkyl-, -C0-C3 alkyl-5-6-membered heteroarylene- C0-C3 alkyl-;R121 is selected from: R134-R135-R136;R134 and R136 are independently selected from: a bond or an optionally substituted group selected from C1-C10 alkylene, C2-C10 alkenylene or C2-C10 alkynylene;R135 is selected from: -C(=O)O-, -OC(=O)-, -OC(=O)O-, -O-, -N(R116)C(=O)-, -C(=O)N(R116)-, - N(R116)C(=O)N(R116)-,-N(R116)C(=O)O-, -OC(=O)N(R116), -S-S- or -C(=O)-;R138 is selected from: R50, R51 or R137; each R50 and R137 is independently defined above or for any aspect or embodiment herein;R51 is selected from:; wherein,R3-R18, R22, R62 through R71, R73 through R79, R105 through R113, R123 through R128, R132, R133, and any remaining variables are as defined above or for any aspect or embodiment herein;R54 is selected from: -CH2-, -C(=O)-, -C(=S)- or -C(NR59)-, -C(=O)O-, -OC(=O)-, -OC(=O)O-, -O-, - N(R116)C(=O)- or -N(R116)C(=O)-O-;R55 is selected from: a bond, or C1-C4 alkylene;R56 is selected from: a bond, -O-, -S-, -S(=O)-, -S(=O)2- or -NR59-;R57 is a bond, C1-C6 alkylene, - C1-C6 alkylene-O-C1-C6 alkylene-;R58 is selected from:R60 is selected from: is absent (i.e., an unbonded pair of electrons on N), -H or optionally substituted C1-C6 alkyl; wherein, when R60 is -H or optionally substituted C1-C6 alkyl, R60 is bound directly to a nitrogen atom with a +1 charge, wherein the +1 charge may be counterbalanced with an anion from a pharmaceutically acceptable acid; each R139 is independently selected from: -H, -OH or C1-C4 alkyl; each R59 is independently selected from: -H, C1-C4 alkyl, C1-C4 cycloalkyl, -(CH2)I-4- C1-C4 cycloalkyl or R31; and,R119 is a 4-7-member heterocyclyl, wherein: one ring atom is a nitrogen atom (nitrogen ring atom) and each remaining ring atom is a carbon atom (carbon ring atom); each carbon ring atom adjacent to the nitrogen ring atom is substituted with R31 to form -CHR31-; each carbon ring atom that is non-adjacent to the nitrogen ring atom is unsubstituted; said heterocyclyl has a ring with 0 or 1 carbon-carbon double bond, i.e., -CH=CH-; and, when R48 is bound to a carbon ring atom, i.e., when the nitrogen ring atom is not bound to R48, then said nitrogen ring atom is substituted with R31.The structural formulas of an ionizable cationic lipid or cationic lipid of the present invention may be further defined for any aspect or embodiment as follows:

2. The compound of claim 1, wherein R1 or R19 is selected from:; wherein, for each R1 or R19 group, one R33 is -R22-R21- (where R21, in the case of R19, is bound to R2) and each remaining R33 is independently selected from: -H, -CH2(halo), -CH(halo)2, - C(halo)3, -F, -CF3, -Cl, -Br, -I, -CH3, -CH2R25, -CH(R25)2, -C(R25)3, -OH, -OR25, -NH2, -NHR25, - N(R25)2, -SH, -SR25, -S(=O)H, -S(=O)R25, -S(=O)2H, -S(=O)2R25, -PH2, -PHR25, -P(R25)2, -P(OH)2, -POHR25, -P(R25)2, -P(OR25)2, -P(O)H2, -P(O)HR25, -P(O)(R25)2, -P(O)(OH)2, -P(O)(OR25)OH, - P(O)(OR25)2, -SiH3, -SiH2R25, -SiH(R25)2or -Si(R25)3.

3. The compound of claim 2, wherein for each R1 or R19 group, one R33 is -R22-R21- and each remaining R33 group is independently selected from: -H, -F, -CF3, -Cl, -Br, -I, -OH, -S(=O)2R25 or - NHR25.

4. The compound of claim 2, wherein R1 or R19 is selected from:wherein, each R33 is independently selected from: -H, -CH2(halo), -CH(halo)2, -C(halo)3, -F, -CF3, -Cl, -Br, -I, -CH3, -CH2R25, -CH(R25)2, -C(R25)3, -OH, -OR25, -NH2, -NHR25, -N(R25)2, -SH, -SR25, -S(=O)H, -S(=O)R25, -S(=O)2H, -S(=O)2R25, -PH2, -PHR25, -P(R25)2, -P(OH)2, -POHR25, -P(R25)2, - P(OR25)2, -P(O)H2, -P(O)HR25, -P(O)(R25)2, -P(O)(OH)2, -P(O)(OR25)OH, -P(O)(OR25)2, -SiH3, - SiH2R25, -SiH(R25)2, or -Si(R25)3.

5. The compound of any one of claims 1-4, wherein R1 is selected from:, , group consisting of: -H, -CHjfhalo), -CH(halo)2, -C(halo)3, -F, -CF3, -Cl, -Br, -I, -CH3, -CH2R25, - CH(R25)2, -C(R25)3, -OH, -OR25, -NH2, -NHR25, -N(R25)2, -SH, -SR25, -S(=O)H, -S(=O)R25, -S(=O)2H, -S(=O)2R25, -S(=O)2NH2, -PH2, -PHR25, -P(R25)2, -P(OH)2, -POHR25, -P(R25)2, -P(OR25)2, -P(O)H2, -P(O)HR25, -P(O)(R25)2, -P(O)(OH)2, -P(O)(OR25)OH, -P(O)(OR25)2, -SiH3, - SiH2R25, -SiH(R25)2, and -Si(R25)3.

6. The compound of claim 5, wherein: each R33 is independently selected from: -H, -F, -CF3, -Cl, -Br, - I, -OH, -S(=O)2CH3, -S(=O)2R25, -NHCH3 or -NHR25 and R22 is selected from a bond, -CH2-, -[CH2]2-, -C(=O)- -CH(OH)- or -NH-.

7. The compound of claim 1, wherein:R1 iswherein,A1through A5, A13and A14are each independently selected from: -CR33=, -C(R33)2-, -N=, - NR33-, -O-, -S- and -S(R46)-; for each R1 or R19, one R33 is -R22-R21- and each remaining R33 is independently selected from the group consisting of: a bond, -H, -CH2(halo), -CH(halo)2, -C(halo)3, -F, -CF3, -Cl, -Br, -I, -CH3, -OCH3, -CH2R25, - CH(R25)2, -C(R25)3, -OH, -OR25, -NH2, -NHR25, -N(R25)2, -SH, -SR25, -S(=O)H, -S(=O)R25, - S(=O)2H, -S(=O)2R25, -PH2, -PHR25, -P(R25)2, -P(OH)2, -POHR25, -P(R25)2, -P(OR25)2, -P(O)H2, - P(O)HR25, -P(O)(R25)2, -P(O)(OH)2, -P(O)(OR25)OH, -P(O)(OR25)2, -SiH3, -SiH2R25, -SiH(R25)2, and -Si(R25)3; and, each ™ is independently selected from a single or double bond, wherein selection of said single or double bond maintains proper valency, i.e., does not cause a valency violation such as a carbon, nitrogen or oxygen atom violating the octet rule.

9. The compound of claim 1, wherein:R1 or R19 is selected from:wherein, for each R1 or R19, one R33 is -R22-R21- (where R21, in the case of Rl, is bound to R2) and each remaining R33 is independently selected from the group consisting of: a bond, -H, - CH2(halo), -CH(halo)2, -C(halo)3, -F, -CF3, -Cl, -Br, -I, -CH3, -CH2R25, -CH(R25)2, -C(R25)3, -OH, - OR25, -NH2, -NHR25, -N(R25)2, -SH, -SR25, -S(=O)H, -S(=O)R25, -S(=O)2H, -S(=O)2R25, -PH2, - PHR25, -P(R25)2, -P(OH)2, -POHR25, -P(R25)2, -P(OR25)2, -P(O)H2, -P(O)HR25, -P(O)(R25)2, - P(O)(OH)2, -P(O)(OR25)OH, -P(O)(OR25)2, -OS(=O)2NHR25-, -S(=O)2NHR25, -OS(=O)2NH2, - S(=O)2NH2, -SiH3, -SiH2R25, -SiH(R25)2, and -Si(R25)3; andeach ~ is independently selected from a single or double bond, wherein selection of said single or double bond maintains proper valency, i.e., does not cause a valency violation such as a carbon, nitrogen or oxygen atom violating the octet rule.

10. The compound of claim 1, wherein:R1 or R19 is selected from:; whereinA1through A14are each independently selected from: -CR33=, -C(R33)2-, -N=, -NR33-, -O-, -S-or -S(R46)-; for each R1 or R19, one R33 is -R22-R21- (where R21, in the case of Rl, is bound to R2) and each remaining R33 is independently selected from: a bond, -H, methyl, ethyl, -CH2(halo), - CH(halo)2, -C(halo)3, -F, -CF3, -Cl, -Br, -I, -CH3, -OCH3, -CH2R25, -CH(R25)2, -C(R25)3, -OH, -OR25, -CN, -NO2, -NH2, -NHR25, -N(R25)2, -SH, -SR25, -S(=O)H, -S(=O)R25, -S(=O)CH3, -S(=O)2H, - S(=O)2R25, -OS(=O)2NH-R25 -S(=O)2NHR25, -PH2, -PHR25, -P(R25)2, -P(OH)2, -POHR25, -P(R25)2, -P(OR25)2, -P(O)H2, -P(O)HR25, -P(O)(R25)2, -P(O)(OH)2, -P(O)(OR25)OH, -P(O)(OR25)2, - OS(=O)2NH2, -S(=O)2NH2, -SiH3, -SiH2R25, -SiH(R25)2, or -Si(R25)3;R46 is selected from: absent, =0 or (=O)2; and each — is independently selected from a single or double bond, wherein selection of said single or double bond maintains proper valency, i.e., does not cause a valency violation such as a carbon, nitrogen or oxygen atom violating the octet rule.

11. The compound of claim 1, wherein: R23 is an optionally substituted group selected from: a 3-18- membered saturated or partially unsaturated carboaryl, a 5-18-membered unsaturated carboaryl, a 3-18-membered saturated or partially unsaturated heteroaryl, or 5-18-membered unsaturated heteroaryl; wherein said heteroaryl contains one or more of heteroatoms independently selected from N, O or S as a ring atom.

12. The compound of claim 11, wherein: R23 is selected from: a 3-14-membered saturated or partially unsaturated carbocyclyl, a 5-14-membered unsaturated carbocyclyl, a 3-14-membered saturated or partially unsaturated heterocyclyl, or 5-14-membered unsaturated heterocyclyl; wherein said heterocyclyl contains one or more of heteroatoms independently selected from N, O or S as a ring atom.

13. The compound of claim 12, wherein R23 is substituted with at least one R144, wherein R144 is independently selected from: -CH3, -CH2CH3,-CH2(halo), -CH(halo)2, -C(halo)3, -F, -CF3, -Cl, -Br, -I, - CH3, -OCH3, -CH2OCH3, -CH2R25, -CH(R25)2, -C(R25)3, -OH, -OR25, -CN, -NO2,-NH2, -NHR25, -N(R25)2, -SH, -SR25, -S(=O)H, -S(=O)R25, -S(=O)2H, -S(=O)2R25, S(=O)2CH3, -OS(=O)2NHR25 -S(=O)2NHR25, PH2, -PHR25, -P(R25)2, -P(OH)2, -POHR25, -P(R25)2, -P(OR25)2, -P(O)H2, -P(O)HR25, -P(O)(R25)2, - P(O)(OH)2, -P(O)(OR25)OH, -P(O)(OR25)2, -OS(=O)2NH2, -S(=O)2NH2, -SiH3, -SiH2R25, -SiH(R25)2, - Si(R25)3 or benzoyloxy-.

14. The compound of claim 13, wherein R144 is independently selected from: -F, -Cl, -Br or benzoyloxy-15. The compound of claim 12, wherein R23 is an optionally substituted group selected from: pyridinyl, pyridazinyl, pyrimidinyl, pyrazinyl, bicyclo(l.l.l)pentanyl, coumarinyl, furanyl, tetrahydrofuranyl, thiophenyl, pyrrolyl, pyrrolidinyl, pyranyl, piperidinyl, piperazinyl, imidazolyl, thiazolyl, 1,2- thiazinyl, 1,3-thiazinyl, 1,4-thiazinyl, dioxanyl, morpholinyl, 1,2-oxathiolanyl, 1,2-oxazolyl, 1,3- oxazolyl, isoxazolyl, silolyl, indolyl (2,3-benzopyrrolyl), isoindolyl, indolizinyl, quinolinyl, isoquinolinyl, carbazolyl, purinyl, aziridinyl, azetidinyl, pyrrolidinyl or piperidinyl.

16. The compound of claim 1, wherein R25 or R23 is an optionally substituted group selected from: 1- oxa-cyclobutan-2-yl, tetrahydrofuran-3-yl, morpholin-4-yl, 2-thiacyclohex-l-yl, 2-oxo-2- thiacyclohex-l-yl, 2,2-dioxo-2-thiacyclohex-l-yl, 4-methyl-piperazin-2-yl, 2-hydroxy-aziridin-l-yl, 3-oxo-l-oxacyclobutan-2-yl, 2,2-dimethyl-tetrahydrofuran-3-yl, 3-carboxy-morpholin-4-yl, and 1- cyclopropyl-4-methyl-piperazin-2-yl, oxetanyl, azetidinyl, thietanyl, tetrahydrofuranyl, dihydrofuranyl, tetrahydrothienyl, dihydrothienyl, imidazolidinyl, oxazolidinyl, imidazolinyl, isoxazolidinyl, pyrrolidinyl, 2-pyrrolinyl, 3-pyrroliny, tetrahydropyranyl, dihydropyranyl, dioxanyl, 1,3-dioxolanyl, 1,4-dithianyl, hexahydropyrimidine, morpholinyl, piperazinyl, piperidinyl, 2H- pyranyl, 4H-pyranyl, pyrazolidinyl, pyrazolinyl, 1,2,3,6-tetrahydropyridinyl, tetrahydrothiopyranyl, l,l-dioxo-hexahydro-lX6-thiopyranyl, l,l-dioxo-lX6-thiomorpholinyl, thiomorpholinyl, thioxanyl, and trithianyl.

17. The compound of claim 1, wherein R23 or R25 is an optionally substituted cycloalkyl selected from: cyclopropyl, cyclobutyl, cyclopentyl, cyclopentenyl, cyclohexyl, cyclohexenyl, cycloheptyl, cyclooctyl, cyclononyl, cyclodecyl or norbornyl.

18. The compound of claim 12, wherein R23 is an optionally substituted heteroaryl selected from: pyrrolyl, thienyl, thiazolyl, imidazolyl, furanyl, indolyl, isoindolyl, oxazolyl, isoxazolyl, benzothiazolyl, benzoxazolyl, quinolinyl, isoquinolinyl, pyridinyl, pyrimidinyl, pyrazinyl, pyridazinyl, triazolyl or tetrazolyl.

19. The compound of claim 12, wherein R23 is an optionally substituted heteroaryl selected from: azepinyl, acridinyl, benzimidazolyl, benzindolyl, 1,3-benzodioxolyl, benzofuranyl, benzooxazolyl, benzo[d]thiazolyl, benzothiadiazolyl, benzo[b][l,4]dioxepinyl, benzo[b][l,4]oxazinyl, 1,4- benzodioxanyl, benzonaphthofuranyl, benzoxazolyl, benzodioxolyl, benzodioxinyl, benzoxazolyl, benzopyranyl, benzopyranonyl, benzofuranyl, benzofuranonyl, benzofurazanyl, benzothiazolyl, benzothienyl(benzothiophenyl), benzothieno[3,2-d]pyrimidinyl, benzotriazolyl, benzo[4,6]imidazo[l,2-a]pyridinyl, carbazolyl, cinnolinyl, cyclopenta[d]pyrimidinyl, 6,7-dihydro- 5H-cyclopenta[4,5]thieno[2,3-d]pyrimidinyl, 5,6-dihydrobenzo[h]quinazolinyl, 5,6- dihydrobenzo[h]cinnolinyl, 6,7-dihydro-5H-benzo[6,7]cyclohepta[l,2-c]pyridazinyl, dibenzofuranyl, dibenzothiophenyl, furanyl, furazanyl, furanonyl, furo[3,2-c]pyridinyl, 5,6,7,8,9,10- hexahydrocycloocta[d]pyrimidinyl, 5,6,7,8,9,10-hexahydrocycloocta[d]pyridazinyl, 5,6,7,8,9,10- hexahydrocycloocta[d]pyridinyl, isothiazolyl, imidazolyl, indazolyl, indolyl, indazolyl, isoindolyl, indolinyl, isoindolinyl, isoquinolyl, indolizinyl, isoxazolyl, isoxazol-3-one, 5,8-methano-5,6,7,8- tetrahydroquinazolinyl, naphthyridinyl, 1,6-naphthyridinonyl, oxadiazolyl, 2-oxoazepinyl, oxazolyl, oxiranyl, 5,6,6a,7,8,9,10,10a-octahydrobenzo[h]quinazolinyl, 1-phenyl-TH-pyrrolyl, phenazinyl, phenothiazinyl, phenoxazinyl, phthalazinyl, pteridinyl, purinyl, pyranyl, pyrrolyl, pyrazolyl, pyrazolo[3,4-d]pyrimidinyl, pyridinyl, pyrido[3,2-d]pyrimidinyl, pyrido[3,4-d]pyrimidinyl, pyrazinyl, pyrimidinyl, pyridazinyl, pyrrolyl, quinazolinyl, quinoxalinyl, quinolinyl, isoquinolinyl, tetrahydroquinolinyl, 5,6,7,8-tetrahydroquinazolinyl, 5,6,7,8-tetrahydrobenzo[4,5]thieno[2,3- d]pyrimidinyl, 6,7,8,9-tetrahydro-5H-cyclohepta[4,5]thieno[2,3-d]pyrimidinyl, 5, 6,7,8- tetrahydropyrido[4,5-c]pyridazinyl, thiazolyl, thiadiazolyl, thiapyranyl, triazolyl, tetrazolyl, triazinyl, thieno[2,3-d]pyrimidinyl, thieno[3,2-d]pyrimidinyl, thieno[2,3-c]pyridinyl or thiophenyl (i.e. thienyl).

20. The compound of claim 12, wherein R23 is selected from:wherein:R35 and R41 are each independently selected from:R36 through R40 and R42 through R45 are each independently selected from -CR24=, - C(R24)2-, -N=, -NR24-, -O-, -S- or -S(R46)-;R46 is selected from: absent (i.e., two lone electron pairs), =0 or (=0)2; and each LZ. is independently selected from a single or double bond, wherein selection of said single or double bond maintains proper valency, i.e., does not cause a valency violation such as a carbon, nitrogen or oxygen atom exceeding the octet valency rule for the period having C, N and O.

21. The compound of claim 12, wherein R23 is selected from:; whereinR35 and R41 are each independently selected from:one or two substituents selected from R36 through R40 and one or two substituents selected from R42 through R45 are each independently selected from -N= or -NR24-; and the remaining substituents selected from R36 through R40 and R42 through R45 are selected from -CR24= or -C(R24)2-; wherein each — is independently selected from a single or double bond, wherein selection of said single or double bond maintains proper valency, i.e., does not cause a valency violation such as a carbon, nitrogen or oxygen atom violating the octet rule.

22. The compound of claim 1, wherein R25 for each occurrence, is independently selected from: -H, or an optionally substituted group selected from: C1-C10 alkyl, C2-Cio alkenyl, C2-Cio alkynyl, 3-14- membered cycloalkyl, 5-14-membered spirocycloalkyl or 3-14-membered heterocycloalkyl.

23. The compound of claim 1, wherein R25, for each occurrence, is independently selected from: -CH3, -CF3, -CH2R114, -CH(R114)2, -CH2(halo), -CH(halo)2, -C(halo)3, -OH, -NH2, -NHR114, -SH, -S(=O)H, - S(=O)2H, -PH2, -PHR114, -P(OH)2, POHR114, -P(OH)R114, -P(O)H2, -P(O)HR114, -P(O)(OH)2, - P(O)(OR114)OH, -P(O)(OR114)2, -SiH3, -SiH2R114 or -SiH(R114)2.

24. The compound of claim 1, wherein R114, for each occurrence, is independently selected from: -H or an optionally substituted, linear or branched, C1-C6 alkyl, C3-Cg alkenyl or C3-Cg alkynyl.

25. The compound of claim 1, wherein, R25, for each occurrence, is independently selected from: -H, -CD3, -CH3, -CH2CH3, -F, -Cl, -Br, -I, -OCH3, -OH or -NH2.

26. The compound of claim 1, wherein, R25 is a group selected from: linear or branched, C1-C10 alkyl, C2-Cio alkenyl; or C2-Ci0alkynyl, C5-Ci6spirocycloalkyl, C5-Ci6-cycloalkyl or C1-C10 heterocycloalkyl; wherein said group is optionally substituted with one or more substituents independently selected from: -CH2(halo), -CH(halo)2, -C(halo)3, -F, -Cl, -Br, -CF3, -I, -CH3, -OH, -NH2, -SH, -S(=O)H, -S(=O)2H, - PH2, -P(OH)2, -P(O)H2, -P(O)(OH)2, -SiH3or R15.

27. The compound of claim 1, wherein R22 is selected from: -CH2-, -[CH2]2-, -[CH2]3-, isopropylene, butylene, isobutylene, -N(R116)-, -C(R145)2-, NHC(=O)O-, -OC(=O)NH-, -P(OH)- or -P(O)OH-, wherein, R116 is -H or Ci-3alkyl, and R145, for each occurrence, is independently selected from: - H, -OCH3, -OH or CI-C6alkyl.

28. The compound of claim 1, wherein R2 is selected from: a bond or an optionally substituted, linear or branched, group selected from: -[CH2]O-3CH=CH[CH2]I.3-, -[CH2]o-3CH=CHCH2-, -[CH2]I. 3CH=CH[CH2]I.3-, -[CH2]0-3C=C[CH2]I-3-, -[CH2]I-3C=C[CH2]I-3-, -CO-C8acyclic hydrocarbylene-CH(R47)- Co-Cg akylene-, -Co-Cg acyclic hydrocarbylene-CH(R47)-[CH2]o-3CH=CH[CH2]i.3, -Co-Cg acyclic hydrocarbylene-CH(R47)-[CH2]o-3CHECH[CH2]i.3-, -Co-Cg akylene-R47-Co-Cg akylene-, -Co-Cg akylene- R146-Co-Cg akylene-, -Co-Cg acyclic hydrocarbylene-R143-Co-Cg akylene-, -Co-Cg acyclic hydrocarbylene-R143-[CH2]o-3CH=CH[CH2]i.3, -Co-Cg acyclic hydrocarbylene-R143-[CH2]o-3CEC[CH2]i. 3-, -Co-Cg akylene-R143-Co-Cg akylene-, -Co-Cg acyclic hydrocarbylene-R146-Co-Cg akylene-, -Co-Cg acyclic hydrocarbylene-R146-[CH2]o-3CH=CH[CH2]i.3, -Co-Cg acyclic hydrocarbylene-R146-[CH2]o- 3CEC[CH2]I.3-, -Co-Cg akylene-R146-C0-Cg akylene-, -C0-C8acyclic hydrocarbylene-CH(R147)-C0-Cg akylene-, -Co-Cg acyclic hydrocarbylene-CH(R147)-[CH2]o-3CH=CH[CH2]i.3, -Co-Cg acyclic hydrocarbylene-CH(R147)-[CH2]o-3CEC[CH2]i.3-, -Co-Cg akylene-R147-Co-Cg akylene-, Co-Cg acyclic hydrocarbylene-O-C1-C6 acyclic hydrocarbylene, -C0-C8acyclic hydrocarbylene-O-C0-C8akylene-, - Co-Cg acyclic hydrocarbylene-O-C1-C6 akylene-, -Co-Cg acyclic hydrocarbylene-0-[CH2]o- 3CH=CH[CH2]I.3, -Co-Cg acyclic hydrocarbylene-0-[CH2]o-3CEC[CH2]i.3-, -Co-Cg akylene-O-Co-Cg akylene-, -C0-C8akylene-O-C1-C6 akylene- or -(CH2)o-6-0-(CH2)0-6-.

29. The compound of claim 1, wherein R2 is a linear, branched or optionally substituted group selected from: Ci-Cg alkylene, C2-C8alkenylene, C2-C8alkynylene, C2-C8internal alkenylene, C2-C8internal alkynylene, -C1-C6 akylene-R47-C1-C6 akylene-, -C1-C6 akylene-R143-C1-C6 akylene-, -C1-C6 akylene- R146-C1-C6 akylene-, -C1-C6 akylene-CH(R147)-C1-C6 akylene-, -C1-C6 akylene-O-C1-C6 akylene-, -Ci- C6akylene-S-C1-C6 akylene-, -C1-C6 akylene-S-S-C1-C6 akylene-, -C1-C6 akylene-N(R114)-Ci-Cs akylene-.

30. The compound of claim 1, wherein R2 is selected from:C1-C6 alkylene, C1-C6 alkenylene, C1-C6 alkynylene, -[CH2]o-6-0-[CH2]o-6- or -[CH2]O-6-0-[CH2]I.6-, -[CH2]I.6-0-[CH2]O-6- , -l,4-piperazin-l,4-diyl- , -l,3-piperazin-l,3-diyl- or -l,4-diazepan-l,4-diyl.

31. The compound of claim 1, wherein R2 or R146 is selected from: C3-C7cycloakylene or C3-C7cycloakenylene.

32. The compound of claim 1, wherein R47, for each occurrence, is a 3-10-membered group independently selected from: cycloalkylene, cycloalkenylene, saturated heterocyclene or partially unsaturated heterocyclene.

33. The compound of claim 32, wherein, R47, for each occurrence, is independently selected from: an optionally substituted 3-8-membered saturated, partially unsaturated or unsaturated carbocyclene or an optionally substituted 3-8-membered saturated, partially unsaturated or unsaturated heterocyclene.

34. The compound of claim 32, wherein, R47, for each occurrence, is independently a 3-7-membered group independently selected from: cycloalkylene, cycloalkenylene, saturated heterocyclene or partially unsaturated heterocyclene.

35. The compound of claim 32, wherein R47, for each occurrence, is independently selected from: cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, cyclopropenyl, cyclobutenyl, cyclopentenyl, cyclohexenyl, 1,3-cyclohexadienyl or 1,4-cyclohexadienyl.

36. The compound of claim 1, wherein each R50 or R51 has a longest linear chain with a total number of carbon atoms selected from: 16, 17, 18, 19, 20, 21 or 22.

37. The compound of claim 1, wherein R3, R4, Rll and R12 are each independently selected from: a bond, -R26-Ci-Cio alkylene-R27-, -R26-C2-CIO alkenylene-R27-, -R26-C2-CIO alkynylene-R27-, -R26- C3-Cio carbocyclylene-R27-, -R26-C3-CIO heterocyclylene-R27-; wherein, R26 and R27 are each independently selected from: a bond, or an optionally substituted C1-C6 alkylene, C2-C6 alkenylene or C2-C6 alkynylene.

38. The compound of claim 1, wherein R3, R4, Rll and R12 are each independently selected from: Ci- Cio alkylene, C2-CM alkenylene or C2-CM alkynylene, C3-CM carbocyclylene or C3-Cio heterocyclylene.

39. The compound of claim 1, wherein R3, R4, Rll and R12 are each independently selected from:, R26-C3-C6-cycloalkylene-R27-, -R26-CH=CH-CH2-CH=CH-R27- , -R26-CH=CH-CH2-R47-R27-, -R26- R47-CH2-CH=CH-R27-, -R26-R47-CH2-R47-R27-, -R26-CH=CH-CD2-CH=CH-R27-, -R26-CH=CH-CD2- R47-R27-, -R26-R47-CD2-CH=CH-R27-, -R26-R47-CD2-R47-R27-, -R26-CH=CH-CH2-cPr-R27-, -R26- cPr-CH2-CH=CH-R27-, -R26-cPr-CH2-cPr-R27-,-R26-CH=CH-CD2-cPr-R27-, -R26-cPr-CD2-CH=CH-R27- or -R26-cPr-CD2-cPr-R27-; wherein, R26 and R27, for each occurrence, is independently selected from: a bond, Ci-Cio alkylene, Ci-Cio alkenylene or Ci-Cio alkynylene; and R47, for each occurrence, is independently Cs-Cg-cycloalkylene.

40. The compound of claim 1, wherein R5, R6, R9 and RIO are each independently selected from the group consisting of: -O-CH2-, -O-CH(OH)2-, -N(OH)-, -NH- or -C(NH[OH])-.

41. The compound of claim 1, wherein R7 and R8 are each independently selected from: a bond, R3, - CH(OH)-, -CH(NH2)-, -C(NH[OH])-, -NH-, -N(OH)-, -C(=O)-O-R115-O-C(=O)-, -P(OH)-, -P(OH)(R25)-, - P(=O)(OH)-, -P(OR25)2-, -P(OR25)-, -P(=O)(OR25)- or R3.

42. The compound of claim 1, wherein, R115, for each occurrence, is each independently an optionally substituted group selected from: linear or branched Ci-Cg alkylene, Cj-Cg alkenylene or Cj-Cg alkynylene; -R26-Cg-Cio heterocyclylene-R27- or -R26-Cg-Cio carbocyclylene-R27-; -R26-C3-Cg- carbocyclylene-R27-, -R26-C3-C6-heterocyclylene-R27-.

43. The compound of claim 1, wherein R15, R16, R17 and R18 are each independently selected from: -H; an optionally substituted linear or branched, Ci-Cio alkyl, C2-C10 alkenyl, or C2-C10 alkynyl; an optionally substituted, C3-C7cycloalkyl, C3-C7cycloalkenyl, C5-Ci0-spirocycloalkyl, C3-Ci0-carbocyclyl, C3-Cio-heterocyclyl, aryl, or heteroaryl; or -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-cPr-CD2-cPr-R28, -R26-CD2-cPr-R28, -R26-cPr-CD2-cPr-R28, -R26-cPr-CD2-cPr-R28, -R26-cPr- CD2-CH=CH-R28, -R26-cPr-CD2-cPr-R28; or -R26-C3-C6cycloakylene-R26-C3-C6cycloakylene-R28; wherein, R28 is selected from: -H, C1-C10 alkyl, C2-C10 alkenyl or C2-C10 alkynyl.

44. The compound of claim 1, wherein R62, R63, R64, R67, R68, R69, R74, R75, R76, R77, R108, R109, R110 and Rill are each independently selected from: a bond, C1-C6 alkylene, C2-C6 alkenylene, C2- C6alkynylene, C3-Cg cycloalkylene, C3-Cg cycloalkenylene, C3-C7-carbocyclylene, C3-C7- heterocyclylene, -R26-CH=CH-C2-C4 alkylene-CH=CH-R27-, -R26-CH=CH-C2-C6alkylene-R47-R27-, - R26-R47-C2-C6alkylene-CH=CH-R27-, -R26-R47-C2-C6alkylene-R47-R27-, -R26-C3-C6-cycloakylene- R27-, -R26-CH=CH-CH2-CH=CH-R27-, -R26-CH=CH-CH2-R47-R27-, -R26-R47-CH2-CH=CH-R27-, -R26- R47-CH2-R47-R27-, -R26-CH=CH-CD2-CH=CH-R27-, -R26-CH=CH-CD2-R47-R27-, -R26-R47-CD2- CH=CH-R27- or -R26-R47-CD2-R47-R27-; wherein, R26 and R27 are each independently selected from: a bond, C1-C10 alkylene, C2-C10 alkenylene or C2-C10 alkynylene; R47, for each occurrence, is independently a 3-10-membered carbocyclylene or a 3-10-membered heterocyclylene; and each double bond or cPr has a cis configuration.

45. The compound of claim 1, wherein R65, R70, R78, R105 and R112 are each independently selected from: a bond, C1-C6 alkylene, C2-C6 alkenylene, C2-C6 alkynylene, C3-Cg cycloalkylene, C3-Cg cycloalkenylene, C5-Ci0-spirocycloalkylene, 3-7 membered carbocyclylene, 3-7 membered heterocyclylene, R26-CH=CH-C2-C4alkylene-CH=CH-R27-, -R26-CH=CH-C2-C4alkylene-R47-R27-, - R26-R47-C2-C6alkylene-CH=CH-R27-, -R26-R47-C2-C6alkylene-R47-R27-, -CH=CH-CH2-CH=CH-, - CH=CH-CH2-R47-, -R47-CH2-CH=CH-, -R47-CH2-R47-, -CH=CH-CD2-CH=CH-, -CH=CH-CD2-R47-, -R47- CD2-CH=CH- or -R47-CD2-R47-; wherein, R47 for each occurrence, is an optionally substituted group independently selected from 3-7-membered carbocyclylene or 3-7-membered heterocyclylene.

46. The compound of claim 1, wherein R66, R71, R79, R106 and R113 are each independently selected from: -H, linear or branched Ci-Cg alkyl, C2-C8 alkenyl, or C2-C8 alkynyl, C5-Cio-spirocycloalkyl, C3-C10- carbocyclyl, C3-C10-heterocyclyl, 5-10 membered aryl or 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-R28; or -R26-C3-C6cycloakylene-R26-C3-C6cycloakylene-R28; wherein R28 is selected from: -H, C1-C10 alkyl, C2-C10 alkenyl or C2-C10 alkynyl.

47. The compound of claim 46, wherein R66, R71, R79, R106 and R113, each double bond or -cPr- has a cis configuration, each R26 is independently selected from: a bond or linear or branched C1-C6 alkylene, C1-C6 alkenylene or C1-C6 alkynylene, and R28 is selected from: -H, C1-C6 alkyl, C2-C6 alkenyl or C2-C6 alkynyl.

8. The compound of claim 1, wherein, for any one or more variable selected from R15, R16, R17, R18, R23, R25, R66, R71, R79, R106 or R113, 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, cycloheptatrienyl, 1H- pyrrolizidinyl, 1,2-dihydroquinolinyl, 1,5-naphthyridinyl, 1,8-naphthyridinyl, lH-indazolyl, 1H- 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, xylyl, furyl, imidazolyl, isothiazolyl, isoxazolyl, oxadiazolyl, oxazolyl, pyrrolyl, pyrazolyl, pyridazinyl, pyridyl (pyridinyl), pyrimidinyl, thiadiazolyl, thienyl, tetrazolyl, thiazolyl, triazolyl, 1,2-thiazinyl, 1,3-thiazinyl, 1,4-thiazinyl, azepinyl, azetidinyl, dioxothiomorpholinyl, imidazolidinyl, morpholinyl, oxanyl, oxazinyl, oxazolidinyl, oxepinyl, oxetanyl, piperazinyl, 2,5-piperazinedionyl,l,4-diasepanyl, piperidinyl, pyranyl, pyrrolidinyl, tetrahydrofuranyl, tetrahydropyranyl, thianyl, thiomorpholinyl, thiopyranyl, 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 orspiro[2.5]octanyl, spiro[4.5]decanyl,9. The compound of claim 1, wherein for any one or more variable selected from R3, R4, Rll, R12, R15, R16, R17, R18, R47, R62, R63, R64, R65, R67, R68, R69, R70, R74, R75, R76, R77, R78, R105, R108, R109, R110, Rill, R112, R115, R146 (for 3-7 membered carbocyclylenes or heterocyclylenes) or R191, said carbocyclylene or heterocyclylene is independently an optionally substituted group selected from: 1,2-dihydroquinolinediyl, 1,5-naphthyridinediyl, 1,8-naphthyridinediyl, 1H- indazolediyl, lH-isochromenediyl, lH-pyrrolizidinediyl, 1-naphthalenediyl, 2H- benzo[b][l,4]oxazinediyl, 2H-benzo[e][l,2]oxazinediyl, 2h-chromenediyl, 2-naphthalenediyl, 4H- quinolizinediyl, adeninediyl, azaindazolediyl, azaindolediyl, benzimidazolediyl, benzo[b]thiophenediyl, benzo[c][l,2,5]thiadiazolediyl, benzo[c]isothiazolediyl, benzo[c]thiophenediyl, benzo[d]isothiazolediyl, benzo[d]isoxazolediyl, benzo[d]oxazolediyl, benzo[d]thiazolediyl, benzofurandiyl, benzenediyl, cinnolinediyl, cumenediyl, decahydroisoquinolinediyl, decahydroquinolinediyl, guaninediyl, indazolediyl, indenediyl, indolediyl, indolinediyl, indolizinediyl, isobenzofurandiyl, isoindolediyl, isoquinolinediyl, phenylene, phthalazinediyl, pteridinediyl, purinediyl, pyrido[2,3-b]pyrazinediyl, pyrido[4,3- d]pyrimidinediyl, pyrimidinediyl, quinazolinediyl, quinolinediyl, quinoxalinediyl, tetrahydroquinolinediyl, toluenediyl, xylenediyl, 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, 2,5-piperazinedion-3,6-diyl piperidinediyl, 1,4-diazepandiyl, pyranyl, pyrrolidindiyl, tetrahydrofurandiyl, tetrahydropyrandiyl, thianediyl, thiomorpholinediyl, thiopyrandiyl, cyclopropylene (cyclopropanediyl), cyclobutylene (cyclobutanediyl), cyclopentylene (cylcopentanediyl), cyclohexylene (cyclohexenediyl), cycloheptylene (cycloheptanediyl), cyclopropenediyl, cyclobutenylenediyl, cyclopentenylenediyl, cyclohexenediyl, cycloheptenediyl, cyclobutadienediyl, cyclopentadienediyl, cyclohexadienediyl, cycloheptadienediyl, cycloheptatrienediyl, 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,wherein, R117 is selected from -NH-, -CH2-,3 is independently selected from -H, -CH2(halo), -CH(halo)2, -C(halo)3, -F, --0CH3.

50. The compound of claim 1, wherein -R3-R5-R7-R9-R11-R13-R15, -R3-R5-R7-R9-R11-R13-R17, -R4- R6-R8-R10-R12-R14-R16, and -R4-R6-R8-R10-R12-R14-R18, each independently have a total number of carbon atoms between 16-22, inclusive and / or a longest linear chain with a total number of carbon atoms between 16-22, inclusive.

51. The compound of claim 1, wherein, together, -R5-R7-R9- and -R6-R8-R10- are each independently selected from: -O-, -OC(=O)-, -C(=O)O-, -OC(=O)O-, -S-C(=O)-, -C(=O)-S-, -S-S-, -C(=O)N(R164)-, - N(R164)C(=O)-, -N(R164)C(=O)N(R164)-, -O-C(=O)C(R164)2C(=O)O-, -C(=O)O-C(R164)2C(=O)O-, -O- C(=O)C(R164)2-O-C(=O)-, -O-C(R164)2C(=O)O-, -O-C(=O)C(R164)2C-O-, -O-C(R164)2-O-C(=O)-, - C(=O)-O-C(R164)2C-O- or -O-C(=O)-O-R191-O-C(=O)O-.

52. The compound of claim 1, wherein R61 is selected from:

53. The compound of claim 1, wherein said compound is represented by a structural formula selected from:

54. The compound of claim 1, wherein R22 is selected from: a bond, optionally substituted Ci.5alkylene, optionally substituted -Ci-Cg alkylenediyl-O-, -CH2-, -[CH2]2-, -[CH2]3-, isopropylene, butylene, isobutylene, -NH-, -N(R116)-, -O- or -S-.

55. The compound of claim 1, wherein R31 and R32 are each independently an optionally substituted group selected from: C1-5 alkyl, C1-5 hydroxyalkyl, C1-5 alkoxyalkyl, C1-5 alkenyl, C1-5 alkynyl, or R31 and R32, together with the nitrogen atom to which they are attached, form an optionally substituted aziridinyl, azetidinyl, pyrrolidinyl or piperidinyl group.

56. The compound of claim 1, wherein R31 and R32 are each independently an optionally substituted group selected from: C1-C4 alkyl, methyl, ethyl, propyl, 2-propyl, cyclopropylmethyl, cyclopropylethyl, cyclopropylpropyl, cyclobutylmethyl, cyclobutylethyl, cyclobutylpropyl, n-butyl, n-pentyl, n-hexyl, hydroxymethyl, 2-hydroxyethyl, 2-hydroxypropyl, 2-hydroxybutyl, 2- hydroxypentyl, 2-hydroxyhexyl, 3-hydroxypropyl, 4-hydroxybutyl, 5-hydroxypentyl, 6- hydroxyhexyl, 2,3-dihydroxypropyl, l,3-dihydroxy-2-propyl, 2,4-dihydroxy-butyl, 2,3,4-trihydroxy- butyl.

57. The compound of claim 1, wherein R31 and R32 are each independently an optionally substituted group selected from: 3-7 membered- cycloalkyl-Ci-C4alkyl-, 3-7 membered-cycloalkenyl-Ci-C4alkyl-, 3-7 membered-heterocycloalkyl-Ci-C4alkyl-, 3-7 membered- heterocycloalkenyl;58. The compound of claim 1, wherein R31 and R32 are each independently an optionally substituted group selected from: methyl, ethyl, propyl, hydroxymethyl, hydroxyethyl or hydroxypropyl or PhgC- 0-[CH2]O-2CH2.

59. . The compound of claim 1, wherein R34 is selected from: a bond, -N(R116)- or -O-.

60. The compound of claim 1, wherein R47 is an optionally substituted group selected from: 1,2- dihydroquinolindiyl, 1,5-naphthyridindiyl, 1,8-naphthyridindiyl, lH-indazoldiyl, lH-isochromendiyl, lH-pyrrolizidindiyl, 1-naphthalendiyl, 2H-benzo[b][l,4]oxazindiyl, 2H-benzo[e][l,2]oxazindiyl, 2h- chromendiyl, 2-naphthalendiyl, 4H-quinolizindiyl, adenindiyl, azaindazoldiyl, azaindoldiyl, benzimidazoldiyl, benzo[b]thiophendiyl, benzo[c][l,2,5]thiadiazoldiyl, benzo[c]isothiazoldiyl, benzo[c]thiophendiyl, benzo[d]isothiazoldiyl, benzo[d]isoxazoldiyl, benzo[d]oxazoldiyl, benzo[d]thiazoldiyl, benzofurandiyl, benzendiyl, cinnolindiyl, cumendiyl, decahydroisoquinolindiyl, decahydroquinolindiyl, guanindiyl, indazoldiyl, indendiyl, indoldiyl, indolindiyl, indolizindiyl, isobenzofurandiyl, isoindoldiyl, isoquinolindiyl, phenylene, phthalazindiyl, pteridindiyl, purindiyl, pyrido[2,3-b]pyrazindiyl, pyrido[4,3-d]pyrimidindiyl, pyrimidindiyl, quinazolindiyl, quinolindiyl, quinoxalindiyl, tetrahydroquinolindiyl, toluendiyl or xylendiyl, furandiyl, imidazoldiyl, isothiazoldiyl, isoxazoldiyl, oxadiazoldiyl, oxazoldiyl, pyrazoldiyl, pyrroldiyl, pyridazindiyl, pyridindiyl, pyrimidindiyl, thiadiazoldiyl, thiendiyl, tetrazoldiyl, thiazoldiyl, triazoldiyl, azepindiyl, azetidindiyl, dioxothiomorpholindiyl, imidazolidindiyl, morpholindiyl, oxandiyl, oxazindiyl, oxazolidindiyl, oxepindiyl, oxetandiyl, piperazindiyl, piperidindiyl, 1,4-diasepandiyl, 1,4-piperazin- 1,4-diyl-, -l,3-piperazin-l,3-diyl-, l,4-diazepan-l,4-diyl, 2,5-piperazinedion-3,6-diyl, pyranyl, pyrrolidindiyl, tetrahydrofurandiyl, tetrahydropyrandiyl, thiandiyl, thiomorpholindiyl or thiopyrandiyl, cyclopropylene (cyclopropandiyl), cyclobutylene (cyclobutandiyl), cyclopentylene (cylcopentandiyl), cyclohexylene (cyclohexendiyl), cycloheptylene (cycloheptandiyl), cyclopropendiyl, cyclobutenylendiyl, cyclopentenylendiyl, cyclohexendiyl, cycloheptendiyl, cyclobutadiendiyl, cyclopentadiendiyl, cyclohexadiendiyl or cycloheptadiendiyl, cycloheptatriendiyl, adamantandiyl, azabicyclo[3.1.0]hexandiyl, 3-azabicyclo[3.1.1]heptandiyl, bicyclo[2.1.1]hexyl, bicyclo[2.2.1]heptyl, bicyclo[2.2.1]heptenyl, 3-oxa-8- azabicyclo[3.2.1]octandiyl, 6-oxa-3-azabicyclo[3.1.1]heptandiyl, 8-oxa-3- azabicyclo[3.2.1]octandiyl, 3-oxa-6-azabicyclo[3.1.1]heptandiyl, 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]hexandiyl, 2- azaspiro[3.3]heptandiyl, 2-oxa-6-azaspiro[3.3]heptandiyl, spiro[2.2]pentanyl, spiro[3.3]heptanyl, spiro[2.3]hexandiyl, spiro[2.5]octandiyl or spiro[4.5]decandiyl.

61. The compound of claim 1, wherein R47 is an optionally substituted group selected from:

62. The compound of claim 1, wherein R47 is substituted with one or more groups independently selected from: -CH3, -CH2CH3,-CH2(halo), -CH(halo)2, -C(halo)3, -F, -CF3, -Cl, -Br, -I, -CH3, -OCH3, - CH2OCH3, -CH2R25, -CH(R25)2, -C(R25)3, -OH, -OR25, -CN, -NO2,-NH2, -NHR25, -N(R25)2, -SH, -SR25, -S(=O)H, -S(=O)R25, -S(=O)2H, -S(=O)2R25, S(=O)2CH3, -PH2, -PHR25, -P(R25)2, -P(OH)2, -POHR25, - P(R25)2, -P(OR25)2, -P(O)H2, -P(O)HR25, -P(O)(R25)2, -P(O)(OH)2, -P(O)(OR25)OH, -P(O)(OR25)2, - SiH3, -SiH2R25, -SiH(R25)2, or -Si(R25)3.

63. The compound of claim 60, wherein R47 is an optionally substituted phenylene.

64. The compound of claim 1, wherein R48 is an optionally substituted group selected from: C2-C8 alkylene, C2-C8 internal alkenylene, C2-C8 internal alkynylene, -Co-C3alkyl-3-6-membered cycloalkylene-C0-C3alkyl- or -C0-C3alkyl-4-6-membered cycloalkenylene- C0-C3alkyl-.

65. The compound of claim 1, wherein R48 is an optionally substituted group selected from: methylene, ethylene, propylene, C3-Cg branched alkylene, -Co-C3alkyl-3-6-membered cycloalkylene-C0-C3alkyl- or C0-C3alkyl-4-6-membered cycloalkenylene- C0-C3alkyl-;66. The compound of claim 1, wherein R48 is an optionally substituted group selected from: Co-C3alkyl- 3-6-membered cycloalkylene-Co-C3alkyl-, wherein said 3-6-membered cycloalkylene is selected from cyclopropylene, cyclobutylene, cyclopentylene or cyclohexylene;67. The compound of claim 1, wherein R48 is an optionally substituted group selected from: methylene, ethylene, propylene, isopropylene, butylene or isobutylene.

68. The compound of claim 1, wherein R57 is selected from: methylene, ethylene, propylene, -[CH2]I. 5-O-[CH2]I-5-, -[CH2]I.3-O-[CH2]I.3, -[CH2]2-O-[CH2]3- or -[CH2]3-O-[CH2]2-.

69. The compound of claim 1, wherein R120 is selected from: a bond, -C(=O)O-, -OC(=O)-, -C(=O)C-, - O-. -C(=O)N(R116)-, -N(R116)C(=O)-.

70. The compound of claim 1, wherein R54 is selected from: -C(=O)-, -OC(=O)- or -NHC(=O)-.

71. The compound of claim 1, wherein R119 is a substituted 4-7-member heterocyclyl selected from: azetidinyl, pyrrolidinyl, piperindinyl or azepanyl.

72. The compound of claim 1, wherein R59 is selected from: -H, -OH, hydroxymethyl, hydroxyethyl, methyl, ethyl, propyl, isopropyl, or cyclopropylmethyl.

73. The compound of claim 1, wherein R134 and R136 are independently selected from: a bond or an optionally substituted group selected from: C1-C6 alkylene, C2-C6 alkenylene or C2-C6 alkynylene.

74. The compound of claim 1, wherein R135 is selected from: -C(=O)O-, -OC(=O)-, -OC(=O)O-, -O-, - or -C(=O)-.

75. The compound of claim 1, wherein R121 is selected from: -C0-C3 alkylene-C(=O)O- C0-C3alkylene-, -C0-C3 alkylene-OC(=0)-Co-C3 alkylene-, -C0-C3 alkylene-OC(=0)0-Co-C3 alkylene-, -C0-C3 alkylene-O- C0-C3 alkylene-, -C0-C3 alkylene-N(R116)C(=O)-C0-C3alkylene-, -C0-C3 alkylene-C(=O)N(R116)-C0-C3alkylene-, -C0-C3alkylene-N(R116)C(=O)N(R116)-C0-C3alkylene-, -C0-C3alkylene-N(R116)C(=O)O-C0- C3 alkylene-, -C0-C3 alkylene-OC(=0)N(R116)-Co-C3 alkylene-, -C0-C3 alkylene-S-S-Co-Ca alkylene- or- C0-C3 alkylene-C(=0)-Co-C3 alkylene-, -C1-C3 alkylene-C(=O)O- C1-C3 alkylene-, -C1-C3 alkylene- OC(=O)-C1-C3 alkylene-, -C1-C3 alkylene-OC(=O)O-C1-C3 alkylene-, -C1-C3 alkylene-O-C1-C3 alkylene-, -C1-C3 alkylene-N(R116)C(=O)-C1-C3 alkylene-, -C1-C3 alkylene-C(=O)N(R116)-C1-C3 alkylene-, -C1-C3 alkylene-N(R116)C(=O)N(R116)-C1-C3 alkylene-, -C1-C3 alkylene-N(R116)C(=O)O-C1-C3 alkylene-, -Ci- C3alkylene-OC(=O)N(R116)-C1-C3 alkylene-, -C1-C3 alkylene-S-S-C1-C3 alkylene- or-C1-C3 alkylene- C(=O)-C1-C3 alkylene-, -C1-C3 alkylene-C(=O)O- C1-C3 alkylene-, -C1-C3 alkylene-OC(=O)-C1-C3 alkylene-, -C1-C3 alkylene-OC(=O)O-C1-C3 alkylene- or -C1-C3 alkylene-O-C1-C3 alkylene-.

76. The compound of claim 1, wherein the compound is represented by a structural formula selected from: (12)-(20), (21) (23), (24), (27), (29), (30) or (31); wherein (where present):R31 and R32 are ethyl, R31 is hydroxyethyl and R32 is methyl, or R31 is PhaC-O-CHj-CHj- and R32 is methyl;R48 is propylene or butylene;R34 is -O- or -NH-;R47 is benzene-l,3-diyl or benzene-l,4-diyl;R22 is methoxy;R120 is -OC(=O)-; andR121 is -CH2OC(=O)OCH2-.

77. The compound of claim 1, wherein the compound is represented by a structural formula selected from: (15), (19), (24), (30), (31); wherein,R32 is Ci-10 hydroxyalkyl; andR121 is -C1-C10 alkylene-OC(=O)O- C1-C10 alkylene-.

78. The compound of claim 1, wherein each R50, R51, R138 and R137 (where applicable) is represented by a structural formula selected from:

79. The compound of claim 1, wherein R50, R51, R138 and R137 (where applicable) is represented by a structural formula selected from:

80. The compound of claim 1, wherein said compound has a chemical formula or is represented by a structural formula selected from:

81. The compound of claim 1, wherein said compound is represented by a structural formula selected from:

82. The compound of claim 1, wherein said compound is represented by a structural formula selected from:

83. A lipid nanoparticle comprising of a compound of any one of claims 1-82 or a pharmaceutically acceptable salt, solvate or cocrystal thereof.

84. The lipid nanoparticle of claim 83, wherein the lipid component further comprises one or more lipids selected from: a neutral lipid, a structural lipid or a polymer conjugated lipid.

85. The lipid nanoparticle of claim 84, wherein the neutral lipid is selected from: 1,2-distearoyl-sn- glycero-3-phosphocholine (DSPC), 1,2-dioleoyl- sn-glycero-3-phosphoethanolamine (DOPE), 1,2- dilinoleoyl-sn-glycero-3- phosphocholine (DLPC), 1,2-dimyristoyl-sn-glycero-phosphocholine (DMPC), 1,2- dioleoyl-sn-glycero-3-phosphocholine (DOPC), l,2-dipalmitoyl-sn-glycero-3- phosphocholine (DPPC), 1,2-diundecanoyl-sn-glycero-phosphocholine (DUPC), 1- palmitoyl-2- oleoyl-sn-glycero-3-phosphocholine (POPC), 1,2-di-O-octadecenyl-sn- glycero-3-phosphocholine (18:0 Diether PC), l-oleoyl-2-cholesterylhemisuccinoyl-sn- glycero-3-phosphocholine (OChemsPC), l-hexadecyl-sn-glycero-3-phosphocholine (C16 Lyso PC), l,2-dilinolenoyl-sn-glycero-3- phosphocholine, l,2-diarachidonoyl-sn-glycero-3-phosphocholine, 1,2-didocosahexaenoyl-sn- glycero-3-phosphocholine, l,2-diphytanoyl-sn-glycero-3-phosphoethanolamine (ME 16.0 PE), 1,2- distearoyl-sn-glycero-3-phosphoethanolamine, l,2-dilinoleoyl-sn-glycero-3- phosphoethanolamine, l,2-dilinolenoyl-sn-glycero-3-phosphoethanolamine, 1,2-diarachidonoyl- sn-glycero-3-phosphoethanolamine, 1 ,2-didocosahexaenoyl-sn-glycero-3-phosphoethanolamine, 1 ,2- dioleoyl-sn-glycero-3-phospho-rac-(l-glycerol) sodium salt (DOPG) or sphingomyelin.

86. The lipid nanoparticle of claim 83, wherein the structural lipid is selected from: cholesterol, fecosterol, sitosterol, campesterol, stigmasterol, brassicasterol, ergosterol, tomatidine, tomatine, ursolic acid and alpha- tocopherol, 5-a-cholestanol (5a-cholestan-3P-ol), 5-p-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, chole decanoate, cholesteryl-(4'-hydroxy)-butyl ether, cholesteryl hemisuccinate, cholest-5-en-3P-yl hydrogen sulfate, 24-methylene-cholesterol sulfate or a combination thereof.

87. The lipid nanoparticle of claim 83, wherein the polymer conjugated lipid is a PEGylated lipid selected from: 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 or PEG- DSPE.

88. The lipid nanoparticle of claim 83, comprising: about 25 mol % to about 60 mol % of a compound of claim 1; 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.

89. The lipid nanoparticle of claim 88, comprising: about 40 to about 50 mol % of a compound of claim 1; about 10 mol % to about 20 mol % neutral lipid; about 35 mol % to about 45 mol % structural lipid; and about 1 mol % to about 2 mol % of PEGylated lipid.

90. The lipid nanoparticle of claim 83, wherein the lipid nanoparticle has a diameter of from about 30 nm to about 160 nm.

91. The lipid nanoparticle of claim 83, wherein the PEGylated lipid has a structural formula represented by any one of structural formulas 4-1 - 4-25.

92. The lipid nanoparticle of any one of claim 83, wherein the lipid nanoparticle further comprises a diagnostic, prophylactic or therapeutic payload selected from a therapeutic nucleic acid (TNA), a protein, a peptide, a carbohydrate or a small molecule.

93. The lipid nanoparticle of claim 92, wherein the TNA is an mRNA or a self-amplifying RNA.

94. A pharmaceutical lipid nanoparticle composition comprising the lipid nanoparticle of claim 83 and a pharmaceutically acceptable carrier, excipient or diluent.

95. A method of delivering a lipid nanoparticle of claim 83 or 94 to a mammalian cell, comprising contacting the cell with the lipid nanoparticle.

96. The method of claim 12, wherein the cell is a cell of a human subject.

97. A method of treating a disease, disorder or condition in a subject in need of such treatment, comprising administering the pharmaceutical composition of claim 94 to the subject to thereby treat the disease, disorder or condition.

98. Use of a lipid nanoparticle of any one of claims 83-94 in the manufacture of a medicament for the treatment of a disease, disorder or condition.

99. The method or use of any one of claims 95-98, wherein the disease, disorder or condition is selected from the group consisting of 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.

100. The method or use according to claim 99, wherein the disease, disorder or condition is cystic fibrosis or other diseases of the lung, hemophilia A, hemophilia B, thalassemia, anemia or other blood disorders, AIDS, Alzheimer's disease, Parkinson's disease, Huntington's disease, amyotrophic lateral sclerosis, epilepsy, and other neurological disorders, cancer, diabetes mellitus, muscular dystrophies (e.g., Duchenne, Becker), Hurler's disease, adenosine deaminase deficiency, metabolic defects or disorders, retinal degenerative diseases or other diseases of the eye), mitochondriopathies (e.g., Leber's hereditary optic neuropathy (LHON), Leigh syndrome, and subacute sclerosing encephalopathy), myopathies (e.g., facioscapulohumeral myopathy (FSHD) and cardiomyopathies), diseases of solid organs (e.g., brain, liver, kidney, heart), metabolic diseases or disorders (e.g., Fabry disease, Gaucher disease, phenylketonuria (PKU), glycogen storage disease); urea cycle diseases or disorders (e.g., ornithine transcarbamoylase (OTC) deficiency); lysosomal storage diseases or disorders (e.g., metachromatic leukodystrophy (MLD), mucopolysaccharidosis Type II (MPSII; Hunter syndrome)); liver diseases or disorders (e.g., progressive familial intrahepatic cholestasis (PFIC); cancers, tumors, or genetic diseases or disorders (e.g., cystic fibrosis).