"good" buffer-based cationic lipids for nucleic acid delivery

Abstract

The present invention provides, in part, "good" buffer-based cationic lipids of Formula (I), and sub-formulas thereof: (I), or a pharmaceutically acceptable salt thereof. The compounds provided herein can be useful for delivery and expression of mRNA and encoded protein, e.g., as a component of liposomal delivery vehicle, and accordingly can be useful for treating various diseases, disorders and conditions, such as those caused by viral or bacterial infection.

Description

GOOD" BUFFER-BASED CATIONIC LIPIDS FOR NUCLEIC ACID DELIVERYCROSS-REFERENCE TO RELATED APPLICATIONS

[0001] This application claims priority to U. S. Provisional Application No. 63 / 778,257, filed March 26, 2025, which is incorporated herein by reference in its entirety.BACKGROUND

[0002] Delivery of nucleic acids has been explored extensively as a potential therapeutic option for certain disease states. In particular, messenger RNA (mRNA) therapy has become an increasingly important option for the prevention and treatment of various diseases (e.g. in the use of vaccines).

[0003] Efficient delivery of liposome-encapsulated nucleic acids remains an active area of research. Liposome-encapsulated nucleic acids can be administered intramuscularly (IM).

[0004] The cationic lipid component of a liposome plays an important role in facilitating effective encapsulation of the nucleic acid during the loading of liposomes. In addition, cationic lipids may play an important role in the efficient release of the nucleic acid cargo from the liposome into the cytoplasm of a target cell. Various cationic lipids suitable for in vivo use have been discovered. However, there remains a need to identify cationic lipids that are effective for delivery of mRNA (e.g., intravenously or intramuscularly, e.g., in vaccines). There also remains a need to identify cationic lipids that can be synthesized efficiently and cheaply without the formation of potentially toxic by-products.

[0005] " Good" buffers (or Good's buffers) are buffering agents for biochemical and biological research that were first selected and described by Norman Good and his colleagues (Good, N. E., et al. (1966) Hydrogen Ion Buffers for Biological Research. Biochemistry 5(2), 467-477). Most biological reactions take place near-neutral pH between 6 and 8. Good therefore reasoned that an ideal buffer for biochemical or biological applications would have a pKa value in this region to provide maximum buffering capacity. Additional selection criteria included high solubility, lack of toxicity, limited interference with biochemical reactions, very low absorbance between 240 nm and 700 nm, enzymatic and hydrolytic stability, minimal changes due to temperature and concentration, limited effects due to ionic or salt composition of the solution, limited interaction with mineral cations, and limited permeability of biological membranes.

[0006] The foregoing characteristics make " Good" buffers exceptionally good starting points for the synthesis of cationic lipids for use in in vivo settings. Many " Good" buffers remain crucial tools in modern laboratories and are therefore readily available at low cost.SUMMARY OF THE INVENTION

[0007] The present invention provides, among other things, a novel class of cationic lipid compounds for in vivo delivery of therapeutic agents, such as nucleic acids. The cores of the cationic lipids of the present invention are based on the " Good" HEPES, HEPPS, and HEPBS buffers (see Table A below). Previous cationic lipids derived from " Good" buffers include those in WO 2022 / 221688 Al and WO 2023 / 198857 Al, incorporated herein by reference. The inventors of the present invention contemplate that the cationic lipids of the present invention with new linkers at the A1position may be associated with beneficial properties such as advantageous stability. The inventors of the present invention have also surprisingly found that certain lipid nanoparticles comprising a cationic lipid of the present invention, exhibit improved properties relative to lipid nanoparticles comprising other cationic lipids derived from " Good" buffers, and are very effective for the intramuscular and / or intravenous delivery of mRNA encapsulated in said lipid nanoparticles. Indeed, lipid nanoparticles comprising the cationic lipids of the present invention have demonstrated high levels of peptide or protein expression when delivering mRNA encoding for said peptide or protein by intramuscular and / or intravenous delivery. For example, lipid nanoparticles comprising cationic lipids of the present invention and encapsulating human erythropoietin (hEPO) mRNA achieved comparable or greater levels of expression of hEPO mRNA when administered to mice by intramuscular delivery to an ester-linked " Good" buffer lipid, Lipid A (see e.g. Table D below).

[0008] The cationic lipids of the present invention also comprise cleavable groups (e.g., esters, amides, disulphides, carbonates, carbamates, carbamides, and thiocarbamates) that are contemplated to improve biodegradability and thus contribute to their favorable safety profile.

[0009] It is contemplated that these compounds are capable of highly effective in vivo intramuscular and / or intravenous delivery of various therapeutic agents and vaccines (e.g., therapeutic agents and / or vaccines for use in Flu, acne, chlamydia, or a disease or disorder induced by infection with P. gingivalis). It is also contemplated that lipid nanoparticles comprising these cationic lipid compounds are capable of highly effective in vivo delivery while maintaining a favorable safety profile. It is also contemplated that lipid nanoparticles comprising these cationic lipid compounds may exhibit degradation in vivo.

[0010] Provided herein are compounds (e.g. compounds of Formulae (I) and sub formulae thereof) which are cationic lipids.[Oil] In an aspect, provided herein are compounds having a structure according to Formula (I):or a pharmaceutically acceptable salt thereof, wherein:A1is selected from a bond,, -O-, -S-, and -(C1-C6alkylene)-, wherein the left hand side of each depicted structure is bound to the -(CH2)a-;Z1is selected fromthe right hand side of each depicted structure is bound to the -(CH2)a-;each R is independently selected from:, wherein each R1is independently selected from optionally substituted alkyl, optionally substituted alkenyl, and optionally substituted alkynyl;, wherein each R2is independently selected from optionally substituted alkyl, optionally substituted alkenyl, optionally substituted alkynyl, and -W1-X1, wherein each W1is independently selected from optionally substituted alkylene and optionally substituted alkenylene, andeach X1is independently selected from -*O-(C=O)-optionally substituted alkyl, -(*C=O)-O- optionally substituted alkyl, -*O-(C=O)-optionally substituted alkenyl, and -(*C=O)-O-optionally substituted alkenyl, wherein the atom marked with a * is connected to W1;(iii), wherein each R3is independently selected from optionally substituted alkyl, optionally substituted alkenyl, and optionally substituted alkynyl; and(iv), wherein each R4is independently selected from optionally substituted alkyl, optionally substituted alkenyl, and optionally substituted alkynyl;(v) optionally substituted alkyl or optionally substituted alkenyl;each R5is selected from hydrogen and optionally substituted (Ci-Cs) alkyl;each R6is selected from optionally substituted (Ci-Cs) alkyl;each a is independently selected from 1, 2, 3, 4, 5, 6, 7, 8, 9 and 10;each b is 1, 2, 3, 4 or 5;each c is 1, 2, 3, 4, or 5;each d is independently selected from 2, 3, 4, 5, 6, 7, 8, 9 and 10;each e is independently selected from 2, 3, 4, 5, 6, 7, 8, 9 and 10; andeach f is independently selected from 2, 3, 4, 5, 6, 7, 8, 9 and 10.

[0012] In another aspect provided herein is a compound selected from those listed in Table 1, or a pharmaceutically acceptable salt thereof.

[0013] In another aspect provided herein is a composition comprising one or more cationic lipids of the present invention or pharmaceutically acceptable salts thereof, one or more non-cationic lipids, one or more cholesterol-based lipids and one or more PEG-modified lipids.

[0014] Also provided herein are compositions of the invention for use in methods of treating or preventing a disease or disorder amenable to treatment or prevention by a peptide or protein encoded by mRNA that is encapsulated within said composition. Methods of treating or preventing a disease or disorder wherein said method comprises administering to a subject in need thereof a composition of the invention and wherein the disease is amenable to treatment or prevention by a peptide or protein encoded by mRNA that is encapsulated within said composition are also provided.BRIEF DESCRIPTION OF DRAWINGS

[0015] FIG. 1 depicts Scheme 1, the reaction scheme for Example 1A.

[0016] FIG.2 depicts Scheme 2, the reaction scheme for Example IB.

[0017] FIG.3 depicts Scheme 3, the reaction scheme for Example 1C.

[0018] FIG.4 depicts Scheme 4, the reaction scheme for Example ID.

[0019] FIG.5 depicts Scheme 5, the reaction scheme for Example IE.

[0020] FIG.6 depicts Scheme 6, the reaction scheme for Example IF.

[0021] FIG.7 depicts Scheme 7, the reaction scheme for Example 1G.

[0022] FIG.8 depicts Scheme 8, the reaction scheme for Example 1H.

[0023] FIG.9 depicts Scheme 9, the reaction scheme for Example 1J.

[0024] FIG. 10 depicts Scheme 10, the reaction scheme for Example IK.

[0025] FIG. 11 depicts Scheme 11, the reaction scheme for Example 1L.

[0026] FIG. 12 depicts Scheme 12, the reaction scheme for Example IM.

[0027] FIG. 13 depicts Scheme 13, the reaction scheme for Example IN.

[0028] FIG. 14 depicts Scheme 14, the reaction scheme for Example 10.

[0029] FIG. 15 depicts Scheme 15, the reaction scheme for Example IP.DETAILED DESCRIPTION OF CERTAIN EMBODIMENTSDefinitions

[0030] In order for the present invention to be more readily understood, certain terms are first defined below. Additional definitions for the following terms and other terms are set forththroughout the specification. The publications and other reference materials cited herein to describe the background of the invention and to provide additional detail regarding its practice are hereby incorporated by reference.

[0031] Amino acid: As used herein, the term "amino acid," in its broadest sense, refers to any compound and / or substance that can be incorporated into a polypeptide chain. In some embodiments, an amino acid has the general structure H2N-C(H)(R)-COOH. In some embodiments, an amino acid is a naturally occurring amino acid. In some embodiments, an amino acid is a synthetic amino acid; in some embodiments, an amino acid is a d-amino acid; in some embodiments, an amino acid is an l-amino acid. " Standard amino acid" refers to any of the twenty standard l-amino acids commonly found in naturally occurring peptides. " Nonstandard amino acid" refers to any amino acid, other than the standard amino acids, regardless of whether it is prepared synthetically or obtained from a natural source. As used herein, "synthetic amino acid" encompasses chemically modified amino acids, including but not limited to salts, amino acid derivatives (such as amides), and / or substitutions. Amino acids, including carboxy- and / or amino-terminal amino acids in peptides, can be modified by methylation, amidation, acetylation, protecting groups, and / or substitution with other chemical groups that can change the peptide's circulating half-life without adversely affecting their activity. Amino acids may participate in a disulfide bond. Amino acids may comprise one or post-translational modifications, such as association with one or more chemical entities (e.g., methyl groups, acetate groups, acetyl groups, phosphate groups, formyl moieties, isoprenoid groups, sulfate groups, polyethylene glycol moieties, lipid moieties, carbohydrate moieties, biotin moieties, etc.). The term "amino acid" is used interchangeably with "amino acid residue," and may refer to a free amino acid and / or to an amino acid residue of a peptide. It will be apparent from the context in which the term is used whether it refers to a free amino acid or a residue of a peptide.

[0032] Animal: As used herein, the term "animal" refers to any member of the animal kingdom. In some embodiments, "animal" refers to humans, at any stage of development. In some embodiments, "animal" refers to non-human animals, at any stage of development. In certain embodiments, the non-human animal is a mammal (e.g., a rodent, a mouse, a rat, a rabbit, a monkey, a dog, a cat, a sheep, a bovine, a primate, and / or a pig). In some embodiments, animals include, but are not limited to, mammals, birds, reptiles, amphibians, fish, insects, and / or worms. In some embodiments, an animal may be a transgenic animal, genetically engineered animal, and / or a clone.

[0033] Approximately or about: As used herein, the term "approximately" or "about," as applied to one or more values of interest, refers to a value that is similar to a stated reference value. In certainembodiments, the term "approximately" or "about" refers to a range of values that fall within 25%, 20%, 19%, 18%, 17%, 16%, 15%, 14%, 13%, 12%, 11%, 10%, 9%, 8%, 7%, 6%, 5%, 4%, 3%, 2%, 1%, or less in either direction (greater than or less than) of the stated reference value unless otherwise stated or otherwise evident from the context (except where such number would exceed 100% of a possible value).

[0034] Biologically active: As used herein, the term "biologically active" refers to a characteristic of any agent that has activity in a biological system, and particularly in an organism. For instance, an agent that, when administered to an organism, has a biological effect on that organism, is considered to be biologically active.

[0035] Delivery: As used herein, the term "delivery" encompasses both local and systemic delivery. For example, delivery of mRNA encompasses situations in which an mRNA is delivered to a target tissue and the encoded protein is expressed and retained within the target tissue (also referred to as "local distribution" or "local delivery"), and situations in which an mRNA is delivered to a target tissue and the encoded protein is expressed and secreted into patient's circulation system (e.g., serum) and systematically distributed and taken up by other tissues (also referred to as "systemic distribution" or "systemic delivery").

[0036] Expression: As used herein, "expression" of a nucleic acid sequence refers to translation of an mRNA into a polypeptide, assemble multiple polypeptides into an intact protein (e.g., enzyme) and / or post-translational modification of a polypeptide or fully assembled protein (e.g., enzyme). In this application, the terms "expression" and "production," and grammatical equivalents thereof, are used interchangeably.

[0037] Functional: As used herein, a "functional" biological molecule is a biological molecule in a form in which it exhibits a property and / or activity by which it is characterized.

[0038] Half-life: As used herein, the term "half-life" is the time required for a quantity such as nucleic acid or protein concentration or activity to fall to half of its value as measured at the beginning of a time period.

[0039] Helper lipid: The term "helper lipid" as used herein refers to any neutral or zwitterionic lipid material including cholesterol. Without wishing to be held to a particular theory, helper lipids may add stability, rigidity, and / or fluidity within lipid bilayers / nanoparticles.

[0040] Improve, increase, or reduce: As used herein, the terms "improve," "increase," or "reduce," or grammatical equivalents, indicate values that are relative to a baseline measurement, such as a measurement in the same individual prior to initiation of the treatment described herein, or a measurement in a control subject (or multiple control subject) in the absence of the treatmentdescribed herein. A "control subject" is a subject afflicted with the same form of disease as the subject being treated, who is about the same age as the subject being treated.

[0041] In Vitro: As used herein, the term "in vitro" refers to events that occur in an artificial environment, e.g., in a test tube or reaction vessel, in cell culture, etc., rather than within a multicellular organism.

[0042] In Vivo: As used herein, the term "in vivo" refers to events that occur within a multi-cellular organism, such as a human and a non-human animal. In the context of cell-based systems, the term may be used to refer to events that occur within a living cell (as opposed to, for example, in vitro systems).

[0043] Liposome: As used herein, the term "liposome" refers to any lamellar, multilamellar, or solid nanoparticle vesicle. Typically, a liposome as used herein can be formed by mixing one or more lipids or by mixing one or more lipids and polymer(s). In some embodiments, a liposome suitable for the present invention contains a cationic lipid(s) and optionally further comprises:(i) non-cationic lipid(s),(ii) cholesterol-based lipid(s), and / or(iii) PEG-modified lipid(s).

[0044] messenger RNA (mRNA): As used herein, the term "messenger RNA (mRNA)" or "mRNA" refers to a polynucleotide that encodes at least one polypeptide. mRNA as used herein encompasses both modified and unmodified RNA. The term "modified mRNA" related to mRNA comprising at least one chemically modified nucleotide. mRNA may contain one or more coding and non-coding regions. mRNA can be purified from natural sources, produced using recombinant expression systems and optionally purified, chemically synthesized, etc. Where appropriate, e.g., in the case of chemically synthesized molecules, mRNA can comprise nucleoside analogs such as analogs having chemically modified bases or sugars, backbone modifications, etc. An mRNA sequence is presented in the 5' to 3' direction unless otherwise indicated. In some embodiments, an mRNA is or comprises natural nucleosides (e.g., adenosine, guanosine, cytidine, uridine); nucleoside analogs (e.g., 2-aminoadenosine, 2-thiothymidine, inosine, pyrrolo-pyrimidine, 3-methyl adenosine, 5-methylcytidine, C5-propynyl-cytidine, C5-propynyl-uridine, 2-aminoadenosine, C5-bromouridine, C5-fluorouridine, C5-iodouridine, C5-propynyl-uridine, C5-propynyl-cytidine, C5-methylcytidine, 2-aminoadenosine, 7-deazaadenosine, 7-deazaguanosine, 8-oxoadenosine, 8-oxoguanosine, 0(6)-methylguanine, and 2-thiocytidine); chemically modified bases; biologically modified bases (e.g., methylated bases); intercalated bases; modified sugars (e.g., 2' -fluororibose, ribose, 2'-deoxyribose, arabinose, and hexose); and / or modified phosphate groups (e.g., phosphorothioates and S'-N-phosphoramidite linkages).

[0045] Nucleic acid: As used herein, the term "nucleic acid," in its broadest sense, refers to any compound and / or substance that is or can be incorporated into a polynucleotide chain. In some embodiments, a nucleic acid is a compound and / or substance that is or can be incorporated into a polynucleotide chain via a phosphodiester linkage. In some embodiments, "nucleic acid" refers to individual nucleic acid residues (e.g., nucleotides and / or nucleosides). In some embodiments, "nucleic acid" refers to a polynucleotide chain comprising individual nucleic acid residues. In some embodiments, "nucleic acid" encompasses RNA as well as single and / or double-stranded DNA and / or cDNA. In some embodiments, "nucleic acid" encompasses ribonucleic acids (RNA), including but not limited to any one or more of interference RNAs (RNAi), small interfering RNA (siRNA), short hairpin RNA (shRNA), antisense RNA (aRNA), messenger RNA (mRNA), modified messenger RNA (mmRNA), long non-coding RNA (IncRNA), micro-RNA (miRNA) multimeric coding nucleic acid (MCNA), polymeric coding nucleic acid (PCNA), guide RNA (gRNA) and CRISPR RNA (crRNA). In some embodiments, "nucleic acid" encompasses deoxyribonucleic acid (DNA), including but not limited to any one or more of single-stranded DNA (ssDNA), double-stranded DNA (dsDNA) and complementary DNA (cDNA). In some embodiments, "nucleic acid" encompasses both RNA and DNA. In embodiments, DNA may be in the form of antisense DNA, plasmid DNA, parts of a plasmid DNA, pre-condensed DNA, a product of a polymerase chain reaction (PCR), vectors (e.g., Pl, PAC, BAC, YAC, artificial chromosomes), expression cassettes, chimeric sequences, chromosomal DNA, or derivatives of these groups. In embodiments, RNA may be in the form of messenger RNA (mRNA), ribosomal RNA (rRNA), signal recognition particle RNA (7 SL RNA or SRP RNA), transfer RNA (tRNA), transfer-messenger RNA (tmRNA), small nuclear RNA (snRNA), small nucleolar RNA (snoRNA), SmY RNA, small Cajal body-specific RNA (scaRNA), guide RNA (gRNA), ribonuclease P (RNase P), Y RNA, telomerase RNA component (TERC), spliced leader RNA (SL RNA), antisense RNA (aRNA or asRNA), cis-natural antisense transcript (cis-NAT), CRISPR RNA (crRNA), long noncoding RNA (IncRNA), micro-RNA (miRNA), piwi-interacting RNA (piRNA), small interfering RNA (siRNA), transacting siRNA (tasiRNA), repeat associated siRNA (rasiRNA), 73K RNA, retrotransposons, a viral genome, a viroid, satellite RNA, or derivatives of these groups. In some embodiments, a nucleic acid is a mRNA encoding a protein such as an enzyme.

[0046] Patient: As used herein, the term "patient" or "subject" refers to any organism to which a provided composition may be administered, e.g., for experimental, diagnostic, prophylactic, cosmetic, and / or therapeutic purposes. Typical patients include animals (e.g., mammals such as mice, rats, rabbits, non-human primates, and / or humans). In some embodiments, a patient is a human. A human includes pre- and post-natal forms.

[0047] Pharmaceutically acceptable: The term "pharmaceutically acceptable," as used herein, refers to substances that, within the scope of sound medical judgment, are suitable for use in contact with the tissues of human beings and animals without excessive toxicity, irritation, allergic response, or other problem or complication, commensurate with a reasonable benefit / risk ratio.

[0048] Pharmaceutically acceptable salt: Pharmaceutically acceptable salts are well known in the art. For example, S. M. Berge et al., describes pharmaceutically acceptable salts in detail in J.Pharmaceutical Sciences (1977) 66:1-19. Pharmaceutically acceptable salts of the compounds of this invention include those derived from suitable inorganic and organic acids and bases. Examples of pharmaceutically acceptable, non-toxic acid addition salts are salts of an amino group formed with inorganic acids such as hydrochloric acid, hydrobromic acid, phosphoric acid, sulfuric acid, and perchloric acid or with organic acids such as acetic acid, oxalic acid, maleic acid, tartaric acid, citric acid, succinic acid, or malonic acid, or by using other methods used in the art such as ion exchange. Other pharmaceutically acceptable salts include adipate, alginate, ascorbate, aspartate, benzenesulfonate, benzoate, bisulfate, borate, butyrate, camphorate, camphorsulfonate, citrate, cyclopentanepropionate, digluconate, dodecylsulfate, ethanesulfonate, formate, fumarate, glucoheptonate, glycerophosphate, gluconate, hemisulfate, heptanoate, hexanoate, hydroiodide, 2-hydroxy-ethanesulfonate, lactobionate, lactate, laurate, lauryl sulfate, malate, maleate, malonate, methanesulfonate, 2-naphthalenesulfonate, nicotinate, nitrate, oleate, oxalate, palmitate, pamoate, pectinate, persulfate, 3-phenylpropionate, phosphate, picrate, pivalate, propionate, stearate, succinate, sulfate, tartrate, thiocyanate, p-toluenesulfonate, undecanoate, valerate salts, and the like. Salts derived from appropriate bases include alkali metal, alkaline earth metal, ammonium and N+(CI-4 alkyl)4 salts. Representative alkali or alkaline earth metal salts include sodium, lithium, potassium, calcium, magnesium, and the like. Further pharmaceutically acceptable salts include, when appropriate, nontoxic ammonium. Quaternary ammonium, and amine cations formed using counterions such as halide, hydroxide, carboxylate, sulfate, phosphate, nitrate, sulfonate, and aryl sulfonate. Further pharmaceutically acceptable salts include salts formed from the quarternization of an amine using an appropriate electrophile, e.g., an alkyl halide, to form a quarternized alkylated amino salt.

[0049] Systemic distribution or delivery: As used herein, the terms "systemic distribution" or "systemic delivery," or grammatical equivalents thereof, refer to a delivery or distribution mechanism or approach that affect the entire body or an entire organism. Typically, systemic distribution or delivery is accomplished via body's circulation system, e.g., blood stream. Compared to the definition of "local distribution or delivery."

[0050] Subject: As used herein, the term "subject" refers to a human or any non-human animal (e.g., mouse, rat, rabbit, dog, cat, cattle, swine, sheep, horse or primate). A human includes pre- and post-natal forms. In many embodiments, a subject is a human being. A subject can be a patient, which refers to a human presenting to a medical provider for diagnosis or treatment of a disease. The term "subject" is used herein interchangeably with "individual" or "patient." A subject can be afflicted with or is susceptible to a disease or disorder but may or may not display symptoms of the disease or disorder.

[0051] Substantially: As used herein, the term "substantially" refers to the qualitative condition of exhibiting total or near-total extent or degree of a characteristic or property of interest. One of ordinary skill in the biological arts will understand that biological and chemical phenomena rarely, if ever, go to completion and / or proceed to completeness or achieve or avoid an absolute result. The term "substantially" is therefore used herein to capture the potential lack of completeness inherent in many biological and chemical phenomena.

[0052] Target tissues: As used herein, the term "target tissues" refers to any tissue that is affected by a disease to be treated. In some embodiments, target tissues include those tissues that display disease-associated pathology, symptom, or feature.

[0053] Therapeutically effective amount: As used herein, the term "therapeutically effective amount" of a therapeutic agent means an amount that is sufficient, when administered to a subject suffering from or susceptible to a disease, disorder, and / or condition, to treat, diagnose, prevent, and / or delay the onset of the symptom(s) of the disease, disorder, and / or condition. It will be appreciated by those of ordinary skill in the art that a therapeutically effective amount is typically administered via a dosing regimen comprising at least one unit dose.

[0054] Treating: As used herein, the term "treat," "treatment," or "treating" refers to any method used to partially or completely alleviate, ameliorate, relieve, inhibit, prevent, delay onset of, reduce severity of and / or reduce incidence of one or more symptoms or features of a particular disease, disorder, and / or condition. Treatment may be administered to a subject who does not exhibit signs of a disease and / or exhibits only early signs of the disease for the purpose of decreasing the risk of developing pathology associated with the disease.Chemical definitions

[0055] Acyl: As used herein, the term "acyl" refers to Rz-(C=O)-, wherein Rzis, for example, any alkyl, alkenyl, alkynyl, heteroalkyl or heteroalkylene.

[0056] Aliphatic: As used herein, the term aliphatic refers to (C1-C50) hydrocarbons and includes both saturated and unsaturated hydrocarbons. An aliphatic may be linear, branched, or cyclic. Forexample, (C1-C20) aliphatics can include (C1-C20) alkyls (e.g., linear or branched (C1-C20) saturated alkyls), (C2-C20) alkenyls (e.g., linear or branched (C4-C20) dienyls, linear or branched (C6-C20) trienyls, and the like), and (C2-C2o)alkynyls (e.g., linear or branched (C2-C20) alkynyls). (C1-C20) aliphatics can include (C3-C20) cyclic aliphatics (e.g., (C3-C2o)cycloalkyls, (C4-C20) cycloalkenyls, or (C8-C20) cycloalkynyls). In certain embodiments, the aliphatic may comprise one or more cyclic aliphatic and / or one or more heteroatoms such as oxygen, nitrogen, or sulfur and may optionally be substituted with one or more substituents such as alkyl, halo, alkoxyl, hydroxy, amino, aryl, ether, ester or amide. An aliphatic group is unsubstituted or substituted with one or more substituent groups as described herein. For example, an aliphatic may be substituted with one or more (e.g., 1, 2, 3, 4, 5, or 6 independently selected substituents) of halogen, -COR", -CO2H, -CO2R", -CN, -OH, -OR", -OCOR", -OCO2R", -NH2, -NHR", -N(R")2, -SR" or -SO2R", wherein each instance of R" independently is (C1-C20) aliphatic (e.g., (C1-C20) alkyl, (C1-C15) alkyl, (C1-C10) alkyl, or (C1-C3) alkyl). In embodiments, R" independently is an unsubstituted alkyl (e.g., unsubstituted (C1-C20) alkyl, (C1-C15) alkyl, (C1-C10) alkyl, or (C1-C3) alkyl). In embodiments, R" independently is unsubstituted (C1-C3) alkyl. In embodiments, the aliphatic is unsubstituted. In embodiments, the aliphatic does not include any heteroatoms.

[0057] Alkyl: As used herein, the term "alkyl" means acyclic linear and branched hydrocarbon groups, e.g. "(C1-C30) alkyl" refers to alkyl groups having 1-30 carbons. An alkyl group may be linear or branched. Examples of alkyl groups include, but are not limited to, methyl, ethyl, n-propyl, isopropyl, butyl, isobutyl, sec-butyl, tert-butyl, pentyl, isopentyl tert-pentylhexyl, isohexyl, etc. The term "lower alkyl" means an alkyl group straight chain or branched alkyl having 1 to 6 carbon atoms. Other alkyl groups will be readily apparent to those of skill in the art given the benefit of the present disclosure. An alkyl group may be unsubstituted or substituted with one or more substituent groups as described herein. For example, an alkyl group may be substituted with one or more (e.g., 1, 2, 3, 4, 5, or 6 independently selected substituents) of halogen, -COR", -CO2H, -CO2R", -CN, -OH, -OR", -OCOR", -OCO2R", -NH2, -NHR", -N(R")2, -SR" or -SO2R", wherein each instance of R" independently is (C1-C20) aliphatic (e.g., (C1-C20) alkyl, (C1-C15) alkyl, (C1-C10) alkyl, or (C1-C3) alkyl). In embodiments, R" independently is an unsubstituted alkyl (e.g., unsubstituted (C1-C20) alkyl, (C1-C15) alkyl, (C1-C10) alkyl, or (C1-C3) alkyl). In embodiments, R" independently is unsubstituted (C1-C3) alkyl. In embodiments, the alkyl is substituted (e.g., with 1, 2, 3, 4, 5, or 6 substituent groups as described herein). In embodiments, an alkyl group is substituted with a -OH group and may also be referred to herein as a "hydroxyalkyl" group, where the prefix denotes the -OH group and "alkyl" is as described herein.

[0058] As used herein, "alkyl" also refers to a radical of a straight-chain or branched saturated hydrocarbon group having from 1 to 50 carbon atoms ("(C1-C50) alkyl"). In some embodiments, an alkyl group has 1 to 40 carbon atoms ("(C1-C40) alkyl"). In some embodiments, an alkyl group has 1 to 30 carbon atoms ("(C1-C30) alkyl"). In some embodiments, an alkyl group has 1 to 20 carbon atoms ("(C1-C20) alkyl"). In some embodiments, an alkyl group has 1 to 10 carbon atoms ("(C1-C10) alkyl"). In some embodiments, an alkyl group has 1 to 9 carbon atoms ("(C1-C9) alkyl"). In some embodiments, an alkyl group has 1 to 8 carbon atoms ("(C1-C8) alkyl"). In some embodiments, an alkyl group has 1 to 7 carbon atoms ("(C1-C7) alkyl"). In some embodiments, an alkyl group has 1 to 6 carbon atoms ("(C1-C6) alkyl"). In some embodiments, an alkyl group has 1 to 5 carbon atoms ("(C1-C5) alkyl"). In some embodiments, an alkyl group has 1 to 4 carbon atoms ("(C1-C4) alkyl"). In some embodiments, an alkyl group has 1 to 3 carbon atoms ("(C1-C3) alkyl"). In some embodiments, an alkyl group has 1 to 2 carbon atoms ("(C1-C2) alkyl"). In some embodiments, an alkyl group has 1 carbon atom ("C1 alkyl"). In some embodiments, an alkyl group has 2 to 6 carbon atoms ("(C2-C6) alkyl"). Examples of (C1-C6) alkyl groups include, without limitation, methyl (C1), ethyl (C2), n-propyl (C3), isopropyl (C3), n-butyl (C4), tert-butyl (C4), sec-butyl (C4), iso-butyl (C4), n-pentyl (C5), 3-pentanyl (C5), amyl (C5), neopentyl (C5), 3-methyl-2-butanyl (C5), tertiary amyl (C5), and n-hexyl (C6). Additional examples of alkyl groups include n-heptyl (C7), n-octyl (C8) and the like. Unless otherwise specified, each instance of an alkyl group is independently unsubstituted (an "unsubstituted alkyl") or substituted (a "substituted alkyl") with one or more substituents. In certain embodiments, the alkyl group is an unsubstituted (C1-C50) alkyl. In certain embodiments, the alkyl group is a substituted (C1-C50) alkyl.

[0059] Affixing the suffix "-ene" to a group indicates the group is a divalent moiety, e.g., arylene is the divalent moiety of aryl, and heteroarylene is the divalent moiety of heteroaryl.

[0060] Alkylene: The term "alkylene," as used herein, represents a saturated divalent straight or branched chain hydrocarbon group and is exemplified by methylene, ethylene, isopropylene and the like. Likewise, the term "alkenylene" as used herein represents an unsaturated divalent straight or branched chain hydrocarbon group having one or more unsaturated carbon-carbon double bonds that may occur in any stable point along the chain, and the term "alkynylene" herein represents an unsaturated divalent straight or branched chain hydrocarbon group having one or more unsaturated carbon-carbon triple bonds that may occur in any stable point along the chain. In certain embodiments, an alkylene, alkenylene, or alkynylene group may comprise one or more cyclic aliphatic and / or one or more heteroatoms such as oxygen, nitrogen, or sulfur and may optionally be substituted with one or more substituents such as alkyl, halo, alkoxyl, hydroxy, amino, aryl, ether, ester or amide. For example, an alkylene, alkenylene, or alkynylene may be substituted with one or more (e.g., 1, 2, 3, 4, 5, or 6 independently selected substituents) of halogen, -COR", -CO2H, -CO2R",-CN, -OH, -OR", -OCOR", -OCO2R", -NH2, -NHR", -N(R")2, -SR" or -SO2R", wherein each instance of R" independently is (C1-C20) aliphatic (e.g., (C1-C20) alkyl, (C1-C15) alkyl, (C1-C10) alkyl, or (C1-C3) alkyl). In embodiments, R" independently is an unsubstituted alkyl (e.g., unsubstituted (C1-C20) alkyl, (C1-C15) alkyl, (C1-C10) alkyl, or (C1-C3) alkyl). In embodiments, R" independently is unsubstituted (C1-C3) alkyl. In certain embodiments, an alkylene, alkenylene, or alkynylene is unsubstituted. In certain embodiments, an alkylene, alkenylene, or alkynylene does not include any heteroatoms.

[0061] Alkenyl: As used herein, "alkenyl" means any linear or branched hydrocarbon chains having one or more unsaturated carbon-carbon double bonds that may occur in any stable point along the chain, e.g. "(C2-C3o) alkenyl" refers to an alkenyl group having 2-30 carbons. For example, an alkenyl group includes prop-2-enyl, but-2-enyl, but-3-enyl, 2-methylprop-2-enyl, hex-2-enyl, hex-5-enyl, 2,3-dimethylbut-2-enyl, and the like. In embodiments, the alkenyl comprises 1, 2, or 3 carbon-carbon double bond. In embodiments, the alkenyl comprises a single carbon-carbon double bond. In embodiments, multiple double bonds (e.g., 2 or 3) are conjugated. An alkenyl group may be unsubstituted or substituted with one or more substituent groups as described herein. For example, an alkenyl group may be substituted with one or more (e.g., 1, 2, 3, 4, 5, or 6 independently selected substituents) of halogen, -COR", -CO2H, -CO2R", -CN, -OH, -OR", -OCOR", -OCO2R", -NH2, -NHR", -N(R")2, -SR" or -SO2R", wherein each instance of R" independently is (C1-C20) aliphatic (e.g., (C1-C20) alkyl, (C1-C15) alkyl, (C1-C10) alkyl, or (C1-C3) alkyl). In embodiments, R" independently is an unsubstituted alkyl (e.g., unsubstituted (C1-C20) alkyl, (C1-C15) alkyl, (C1-C10) alkyl, or (C1-C3) alkyl). In embodiments, R" independently is unsubstituted (C1-C3) alkyl. In embodiments, the alkenyl is unsubstituted. In embodiments, the alkenyl is substituted (e.g., with 1, 2, 3, 4, 5, or 6 substituent groups as described herein). In embodiments, an alkenyl group is substituted with a-OH group and may also be referred to herein as a "hydroxyalkenyl" group, where the prefix denotes the -OH group and "alkenyl" is as described herein.

[0062] As used herein, "alkenyl" also refers to a radical of a straight-chain or branched hydrocarbon group having from 2 to 50 carbon atoms and one or more carbon-carbon double bonds (e.g., 1, 2, 3, or 4 double bonds) ("(C2-C5o) alkenyl"). In some embodiments, an alkenyl group has 2 to 40 carbon atoms ("(C2-C4o) alkenyl"). In some embodiments, an alkenyl group has 2 to 30 carbon atoms ("(C2-C30) alkenyl"). In some embodiments, an alkenyl group has 2 to 20 carbon atoms ("(C2-C2o) alkenyl"). In some embodiments, an alkenyl group has 2 to 10 carbon atoms ("(C2-C10) alkenyl"). In some embodiments, an alkenyl group has 2 to 9 carbon atoms ("(C2-C9) alkenyl"). In some embodiments, an alkenyl group has 2 to 8 carbon atoms ("(C2-C8) alkenyl"). In some embodiments, an alkenyl group has 2 to 7 carbon atoms ("(C2-C7) alkenyl"). In some embodiments, an alkenyl group has 2 to 6 carbon atoms ("(C2-C6) alkenyl"). In some embodiments, an alkenyl group has 2 to 5 carbon atoms("(C2-C5) alkenyl"). In some embodiments, an alkenyl group has 2 to 4 carbon atoms ("(C2-C4) alkenyl"). In some embodiments, an alkenyl group has 2 to 3 carbon atoms ("(C2-C3) alkenyl"). In some embodiments, an alkenyl group has 2 carbon atoms ("(C2) alkenyl"). The one or more carboncarbon double bonds can be internal (such as in 2-butenyl) or terminal (such as in 1-butenyl).Examples of (C2-C4) alkenyl groups include, without limitation, ethenyl (C2), 1-propenyl (C3), 2-propenyl (C3), 1-butenyl (C4), 2-butenyl (C4), butadienyl (C4), and the like. Examples of (C2-Cs) alkenyl groups include the aforementioned (C2-C4) alkenyl groups as well as pentenyl (C5), pentadienyl (C5), hexenyl (C6), and the like. Additional examples of alkenyl include heptenyl (C7), octenyl (C8), octatrienyl (C8), and the like. Unless otherwise specified, each instance of an alkenyl group is independently unsubstituted (an "unsubstituted alkenyl") or substituted (a "substituted alkenyl") with one or more substituents. In certain embodiments, the alkenyl group is an unsubstituted (C2-C5o) alkenyl. In certain embodiments, the alkenyl group is a substituted (C2-C50) alkenyl.

[0063] Alkynyl: As used herein, "alkynyl" means any hydrocarbon chain of either linear or branched configuration, having one or more carbon-carbon triple bonds occurring in any stable point along the chain, e.g., "(C2-C30) alkynyl", refers to an alkynyl group having 2-30 carbons. Examples of an alkynyl group include prop-2-ynyl, but-2-ynyl, but-3-ynyl, pent-2-ynyl, 3-methylpent-4-ynyl, hex-2-ynyl, hex-5-ynyl, etc. In embodiments, an alkynyl comprises one carbon-carbon triple bond. An alkynyl group may be unsubstituted or substituted with one or more substituent groups as described herein. For example, an alkynyl group may be substituted with one or more (e.g., 1, 2, 3, 4, 5, or 6 independently selected substituents) of halogen, -COR", -CO2H, -CO2R", -CN, -OH, -OR", -OCOR", -OCO2R", -NH2, -NHR", -N(R")2, -SR" or -SO2R", wherein each instance of R" independently is (C1-C20) aliphatic (e.g., (C1-C20) alkyl, (C1-C15) alkyl, (C1-C10) alkyl, or (C1-C3) alkyl). In embodiments, R" independently is an unsubstituted alkyl (e.g., unsubstituted (C1-C20) alkyl, (C1-C15) alkyl, (C1-C10) alkyl, or (C1-C3) alkyl). In embodiments, R" independently is unsubstituted (C1-C3) alkyl. In embodiments, the alkynyl is unsubstituted. In embodiments, the alkynyl is substituted (e.g., with 1, 2, 3, 4, 5, or 6 substituent groups as described herein).

[0064] As used herein, "alkynyl" also refers to a radical of a straight-chain or branched hydrocarbon group having from 2 to 50 carbon atoms and one or more carbon-carbon triple bonds (e.g., 1, 2, 3, or 4 triple bonds) and optionally one or more double bonds (e.g., 1, 2, 3, or 4 double bonds) ("(C2-C50) alkynyl"). An alkynyl group that has one or more triple bonds, and one or more double bonds is also referred to as an "ene-yne". In some embodiments, an alkynyl group has 2 to 40 carbon atoms ("(C2-C40) alkynyl"). In some embodiments, an alkynyl group has 2 to 30 carbon atoms ("(C2-C30) alkynyl"). In some embodiments, an alkynyl group has 2 to 20 carbon atoms ("(C2-C20) alkynyl"). In some embodiments, an alkynyl group has 2 to 10 carbon atoms ("(C2-C10) alkynyl"). In some embodiments,an alkynyl group has 2 to 9 carbon atoms ("(C2-C9) alkynyl"). In some embodiments, an alkynyl group has 2 to 8 carbon atoms ("(C2-C8) alkynyl"). In some embodiments, an alkynyl group has 2 to 7 carbon atoms ("(C2-C7) alkynyl"). In some embodiments, an alkynyl group has 2 to 6 carbon atoms ("(C2-C6) alkynyl"). In some embodiments, an alkynyl group has 2 to 5 carbon atoms ("(C2-C5) alkynyl"). In some embodiments, an alkynyl group has 2 to 4 carbon atoms ("(C2-C4) alkynyl"). In some embodiments, an alkynyl group has 2 to 3 carbon atoms ("(C2-C3) alkynyl"). In some embodiments, an alkynyl group has 2 carbon atoms ("(C2) alkynyl"). The one or more carboncarbon triple bonds can be internal (such as in 2-butynyl) or terminal (such as in 1-butynyl).Examples of (C2-C4) alkynyl groups include, without limitation, ethynyl (C2), 1-propynyl (C3), 2-propynyl (C3), 1-butynyl (C4), 2-butynyl (C4), and the like. Examples of (C2-C6) alkynyl groups include the aforementioned (C2-C4) alkynyl groups as well as pentynyl (C5), hexynyl (C6), and the like.Additional examples of alkynyl include heptynyl (C7), octynyl (C8), and the like. Unless otherwise specified, each instance of an alkynyl group is independently unsubstituted (an "unsubstituted alkynyl") or substituted (a "substituted alkynyl") with one or more substituents. In certain embodiments, the alkynyl group is an unsubstituted (C2-C50) alkynyl. In certain embodiments, the alkynyl group is a substituted (C2-C50) alkynyl.

[0065] Aryl: The term "aryl" used alone or as part of a larger moiety as in "aralkyl," refers to a monocyclic, bicyclic, or tricyclic carbocyclic ring system having a total of six to fourteen ring members, wherein said ring system has a single point of attachment to the rest of the molecule, at least one ring in the system is aromatic and wherein each ring in the system contains 4 to 7 ring members. In embodiments, an aryl group has 6 ring carbon atoms ("(Cs) aryl," e.g., phenyl). In some embodiments, an aryl group has 10 ring carbon atoms ("(C10) aryl," e.g., naphthyl such as 1-naphthyl and 2-naphthyl). In some embodiments, an aryl group has 14 ring carbon atoms ("(C14) aryl," e.g., anthracyl). " Aryl" also includes ring systems wherein the aryl ring, as defined above, is fused with one or more carbocyclyl or heterocyclyl groups wherein the radical or point of attachment is on the aryl ring, and in such instances, the number of carbon atoms continue to designate the number of carbon atoms in the aryl ring system. Exemplary aryls include phenyl, naphthyl, and anthracene.

[0066] As used herein, "aryl" also refers to a radical of a monocyclic or polycyclic (e.g., bicyclic or tricyclic) 4n+2 aromatic ring system (e.g., having 6, 10, or 14 n electrons shared in a cyclic array) having 6-14 ring carbon atoms and zero heteroatoms provided in the aromatic ring system ("(C6-C14) aryl"). In some embodiments, an aryl group has 6 ring carbon atoms ("(C6) aryl"; e.g., phenyl). In some embodiments, an aryl group has 10 ring carbon atoms ("(C10) aryl"; e.g., naphthyl such as 1-naphthyl and 2-naphthyl). In some embodiments, an aryl group has 14 ring carbon atoms ("(C14) aryl"; e.g., anthracyl). " Aryl" also includes ring systems wherein the aryl ring, as defined above, isfused with one or more carbocyclyl or heterocyclyl groups wherein the radical or point of attachment is on the aryl ring, and in such instances, the number of carbon atoms continue to designate the number of carbon atoms in the aryl ring system. Unless otherwise specified, each instance of an aryl group is independently unsubstituted (an "unsubstituted aryl") or substituted (a "substituted aryl") with one or more substituents. In certain embodiments, the aryl group is an unsubstituted (C6-C14) aryl. In certain embodiments, the aryl group is a substituted (C6-C14) aryl.

[0067] Arylene: The term "arylene" as used herein refers to an aryl group that is divalent (that is, having two points of attachment to the molecule). Exemplary arylenes include phenylene (e.g., unsubstituted phenylene or substituted phenylene).

[0068] Carbocyclyl: As used herein, "carbocyclyl" or "carbocyclic" refers to a radical of a nonaromatic cyclic hydrocarbon group having from 3 to 10 ring carbon atoms ("(C3-C10) carbocyclyl") and zero heteroatoms in the non-aromatic ring system. In some embodiments, a carbocyclyl group has 3 to 8 ring carbon atoms ("(C3-C8) carbocyclyl"). In some embodiments, a carbocyclyl group has 3 to 7 ring carbon atoms ("(C3-C7) carbocyclyl"). In some embodiments, a carbocyclyl group has 3 to 6 ring carbon atoms ("(C3-C6) carbocyclyl"). In some embodiments, a carbocyclyl group has 4 to 6 ring carbon atoms ("(C4-Cs) carbocyclyl"). In some embodiments, a carbocyclyl group has 5 to 6 ring carbon atoms ("(C5-Cs) carbocyclyl"). In some embodiments, a carbocyclyl group has 5 to 10 ring carbon atoms ("(C5-C10) carbocyclyl"). Exemplary (C3-C6) carbocyclyl groups include, without limitation, cyclopropyl (C3), cyclopropenyl (C3), cyclobutyl (C4), cyclobutenyl (C4), cyclopentyl (C5), cyclopentenyl (C5), cyclohexyl (Cs), cyclohexenyl (Cs), cyclohexadienyl (Cs), and the like. Exemplary (C3-C8) carbocyclyl groups include, without limitation, the aforementioned (C3-C6) carbocyclyl groups as well as cycloheptyl (C7), cycloheptenyl (C7), cycloheptadienyl (C7), cycloheptatrienyl (C7), cyclooctyl (Cs), cyclooctenyl (Cs), bicyclo[2.2.1]heptanyl (C7), bicyclo[2.2.2]octanyl (Cs), and the like. Exemplary (C3-C10) carbocyclyl groups include, without limitation, the aforementioned (C3-C8) carbocyclyl groups as well as cyclononyl (C9), cyclononenyl (C9), cyclodecyl (C10), cyclodecenyl (C10), octahydro-1H-indenyl (C9), decahydronaphthalenyl (C10), spiro[4.5]decanyl (C10), and the like. As the foregoing examples illustrate, in certain embodiments, the carbocyclyl group is either monocyclic ("monocyclic carbocyclyl") or polycyclic (e.g., containing a fused, bridged or spiro ring system such as a bicyclic system ("bicyclic carbocyclyl") or tricyclic system ("tricyclic carbocyclyl")) and can be saturated or can contain one or more carbon-carbon double or triple bonds." Carbocyclyl" also includes ring systems wherein the carbocyclyl ring, as defined above, is fused with one or more aryl or heteroaryl groups wherein the point of attachment is on the carbocyclyl ring, and in such instances, the number of carbons continue to designate the number of carbons in the carbocyclic ring system. Unless otherwise specified, each instance of a carbocyclyl group isindependently unsubstituted (an "unsubstituted carbocyclyl") or substituted (a "substituted carbocyclyl") with one or more substituents. In certain embodiments, the carbocyclyl group is an unsubstituted C3-C10 carbocyclyl. In certain embodiments, the carbocyclyl group is a substituted (C3-C10) carbocyclyl.

[0069] In some embodiments, "carbocyclyl" or "carbocyclic" is referred to as a "cycloalkyl", i.e., a monocyclic, saturated carbocyclyl group having from 3 to 10 ring carbon atoms ("(C3-C10) cycloalkyl"). In some embodiments, a cycloalkyl group has 3 to 8 ring carbon atoms ("(C3-C8) cycloalkyl"). In some embodiments, a cycloalkyl group has 3 to 6 ring carbon atoms ("(C3-C6), cycloalkyl"). In some embodiments, a cycloalkyl group has 4 to 6 ring carbon atoms ("(C4-C6) cycloalkyl"). In some embodiments, a cycloalkyl group has 5 to 6 ring carbon atoms ("(C5-C6) cycloalkyl"). In some embodiments, a cycloalkyl group has 5 to 10 ring carbon atoms ("(C5-C10) cycloalkyl"). Examples of (C5-Cs) cycloalkyl groups include cyclopentyl (C5) and cyclohexyl (Cs). Examples of (C3-C6) cycloalkyl groups include the aforementioned (C5-C6) cycloalkyl groups as well as cyclopropyl (C3) and cyclobutyl (C4). Examples of (C3-C8) cycloalkyl groups include the aforementioned (C3-C6) cycloalkyl groups as well as cycloheptyl (C7) and cyclooctyl (C8). Unless otherwise specified, each instance of a cycloalkyl group is independently unsubstituted (an "unsubstituted cycloalkyl") or substituted (a "substituted cycloalkyl") with one or more substituents. In certain embodiments, the cycloalkyl group is an unsubstituted (C3-C10) cycloalkyl. In certain embodiments, the cycloalkyl group is a substituted (C3-C10) cycloalkyl.

[0070] Heteroalkyl: The term "heteroalkyl" is meant a branched or unbranched alkyl, alkenyl, or alkynyl group having from 1 to 14 carbon atoms in addition to 1, 2, 3 or 4 heteroatoms independently selected from the group consisting of N, O, S, and P. Heteroalkyls include tertiary amines, secondary amines, ethers, thioethers, amides, thioamides, carbamates, thiocarbamates, hydrazones, imines, phosphodiesters, phosphoramidates, sulfonamides, and disulfides. A heteroalkyl group may optionally include monocyclic, bicyclic, or tricyclic rings, in which each ring desirably has three to six members. Examples of heteroalkyls include polyethers, such as methoxymethyl and ethoxyethyl.

[0071] Heteroalkylene: The term "heteroalkylene," as used herein, represents a divalent form of a heteroalkyl group as described herein.

[0072] Heteroaryl: The term "heteroaryl," as used herein, represents a fully unsaturated heteroatom-containing ring wherein at least one ring atom is a heteroatom such as, but not limited to, nitrogen and oxygen.

[0073] As used herein, "heteroaryl" also refers to a radical of a 5-14 membered monocyclic or polycyclic (e.g., bicyclic or tricyclic) 4n+2 aromatic ring system (e.g., having 6, 10, or 14 n electronsshared in a cyclic array) having ring carbon atoms and 1 or more (e.g., 1, 2, 3, or 4 ring heteroatoms) ring heteroatoms provided in the aromatic ring system, wherein each heteroatom is independently selected from oxygen, sulfur, nitrogen, boron, silicon, and phosphorus ("5-14 membered heteroaryl"). In heteroaryl groups that contain one or more nitrogen atoms, the point of attachment can be a carbon or nitrogen atom, as valency permits. Heteroaryl polycyclic ring systems can include one or more heteroatoms in one or both rings. " Heteroaryl" includes ring systems wherein the heteroaryl ring, as defined above, is fused with one or more carbocyclyl or heterocyclyl groups wherein the point of attachment is on the heteroaryl ring, and in such instances, the number of ring members continue to designate the number of ring members in the heteroaryl ring system." Heteroaryl" also includes ring systems wherein the heteroaryl ring, as defined above, is fused with one or more aryl groups wherein the point of attachment is either on the aryl or heteroaryl ring, and in such instances, the number of ring members designates the number of ring members in the fused polycyclic (aryl / heteroaryl) ring system. Polycyclic heteroaryl groups wherein one ring does not contain a heteroatom (e.g., indolyl, quinolinyl, carbazolyl, and the like) the point of attachment can be on either ring, i.e., either the ring bearing a heteroatom (e.g., 2-indolyl) or the ring that does not contain a heteroatom (e.g., 5-indolyl).

[0074] In some embodiments, a heteroaryl group is a 5-10 membered aromatic ring system having ring carbon atoms and 1 or more (e.g., 1, 2, 3, or 4) ring heteroatoms provided in the aromatic ring system, wherein each heteroatom is independently selected from oxygen, sulfur, nitrogen, boron, silicon, and phosphorus ("5-10 membered heteroaryl"). In some embodiments, a heteroaryl group is a 5-8 membered aromatic ring system having ring carbon atoms and 1 or more (e.g., 1, 2, 3, or 4) ring heteroatoms provided in the aromatic ring system, wherein each heteroatom is independently selected from oxygen, sulfur, nitrogen, boron, silicon, and phosphorus ("5-8 membered heteroaryl"). In some embodiments, a heteroaryl group is a 5-6 membered aromatic ring system having ring carbon atoms and 1 or more (e.g., 1, 2, 3, or 4) ring heteroatoms provided in the aromatic ring system, wherein each heteroatom is independently selected from oxygen, sulfur, nitrogen, boron, silicon, and phosphorus ("5-6 membered heteroaryl"). In some embodiments, the 5-6 membered heteroaryl has 1 or more (e.g., 1, 2, or 3) ring heteroatoms selected from oxygen, sulfur, nitrogen, boron, silicon, and phosphorus. In some embodiments, the 5-6 membered heteroaryl has 1 or 2 ring heteroatoms selected from oxygen, sulfur, nitrogen, boron, silicon, and phosphorus. In some embodiments, the 5-6 membered heteroaryl has 1 ring heteroatom selected from oxygen, sulfur, nitrogen, boron, silicon, and phosphorus. Unless otherwise specified, each instance of a heteroaryl group is independently unsubstituted (an "unsubstituted heteroaryl") or substituted (a "substituted heteroaryl") with one or more substituents. In certain embodiments, the heteroaryl group is anunsubstituted 5-14 membered heteroaryl. In certain embodiments, the heteroaryl group is a substituted 5-14 membered heteroaryl.

[0075] Exemplary 5-membered heteroaryl groups containing 1 heteroatom include, without limitation, pyrrolyl, furanyl and thiophenyl. Exemplary 5-membered heteroaryl groups containing 2 heteroatoms include, without limitation, imidazolyl, pyrazolyl, oxazolyl, isoxazolyl, thiazolyl, and isothiazolyl. Exemplary 5-membered heteroaryl groups containing 3 heteroatoms include, without limitation, triazolyl, oxadiazolyl, and thiadiazolyl. Exemplary 5-membered heteroaryl groups containing 4 heteroatoms include, without limitation, tetrazolyl. Exemplary 6-membered heteroaryl groups containing 1 heteroatom include, without limitation, pyridinyl. Exemplary 6-membered heteroaryl groups containing 2 heteroatoms include, without limitation, pyridazinyl, pyrimidinyl, and pyrazinyl. Exemplary 6-membered heteroaryl groups containing 3 or 4 heteroatoms include, without limitation, triazinyl and tetrazinyl, respectively. Exemplary 7-membered heteroaryl groups containing 1 heteroatom include, without limitation, azepinyl, oxepinyl, and thiepinyl. Exemplary 5,6-bicyclic heteroaryl groups include, without limitation, indolyl, isoindolyl, indazolyl, benzotriazolyl, benzothiophenyl, isobenzothiophenyl, benzofuranyl, benzoisofuranyl, benzimidazolyl, benzoxazolyl, benzisoxazolyl, benzoxadiazolyl, benzthiazolyl, benzisothiazolyl, benzthiadiazolyl, indolizinyl, and purinyl. Exemplary 6,6-bicyclic heteroaryl groups include, without limitation, naphthyridinyl, pteridinyl, quinolinyl, isoquinolinyl, cinnolinyl, quinoxalinyl, phthalazinyl, and quinazolinyl. Exemplary tricyclic heteroaryl groups include, without limitation, phenanthridinyl, dibenzofuranyl, carbazolyl, acridinyl, phenothiazinyl, phenoxazinyl and phenazinyl.

[0076] As used herein, "heterocyclyl" or "heterocyclic" refers to a radical of a 3- to 14-membered non-aromatic ring system having ring carbon atoms and 1 or more (e.g., 1, 2, 3, or 4) ring heteroatoms, wherein each heteroatom is independently selected from oxygen, sulfur, nitrogen, boron, silicon, and phosphorus ("3-14 membered heterocyclyl"). In heterocyclyl groups that contain one or more nitrogen atoms, the point of attachment can be a carbon or nitrogen atom, as valency permits. A heterocyclyl group can either be monocyclic ("monocyclic heterocyclyl") or polycyclic (e.g., a fused, bridged or spiro ring system such as a bicyclic system ("bicyclic heterocyclyl") or tricyclic system ("tricyclic heterocyclyl")) and can be saturated or can contain one or more carboncarbon double or triple bonds. Heterocyclyl polycyclic ring systems can include one or more heteroatoms in one or both rings. " Heterocyclyl" also includes ring systems wherein the heterocyclyl ring, as defined above, is fused with one or more carbocyclyl groups wherein the point of attachment is either on the carbocyclyl or heterocyclyl ring, or ring systems wherein the heterocyclyl ring, as defined above, is fused with one or more aryl or heteroaryl groups, wherein the point of attachment is on the heterocyclyl ring, and in such instances, the number of ring members continueto designate the number of ring members in the heterocyclyl ring system. Unless otherwise specified, each instance of heterocyclyl is independently unsubstituted (an "unsubstituted heterocyclyl") or substituted (a "substituted heterocyclyl") with one or more substituents. In certain embodiments, the heterocyclyl group is an unsubstituted 3-14 membered heterocyclyl. In certain embodiments, the heterocyclyl group is a substituted 3-14 membered heterocyclyl.

[0077] In some embodiments, a heterocyclyl group is a 5-10 membered non-aromatic ring system having ring carbon atoms and 1 or more (e.g., 1, 2, 3, or 4) ring heteroatoms, wherein each heteroatom is independently selected from oxygen, sulfur, nitrogen, boron, silicon, and phosphorus ("5-10 membered heterocyclyl"). In some embodiments, a heterocyclyl group is a 5-8 membered non-aromatic ring system having ring carbon atoms and 1 or more (e.g., 1, 2, 3, or 4) ring heteroatoms, wherein each heteroatom is independently selected from oxygen, sulfur, nitrogen, boron, silicon, and phosphorus ("5-8 membered heterocyclyl"). In some embodiments, a heterocyclyl group is a 5-6 membered non-aromatic ring system having ring carbon atoms and 1 or more (e.g., 1, 2, 3, or 4) ring heteroatoms, wherein each heteroatom is independently selected from oxygen, sulfur, nitrogen, boron, silicon, and phosphorus ("5-6 membered heterocyclyl"). In some embodiments, the 5-6 membered heterocyclyl has 1 or more (e.g., 1, 2, or 3) ring heteroatoms selected from oxygen, sulfur, nitrogen, boron, silicon, and phosphorus. In some embodiments, the 5-6 membered heterocyclyl has 1 or 2 ring heteroatoms selected from oxygen, sulfur, nitrogen, boron, silicon, and phosphorus. In some embodiments, the 5-6 membered heterocyclyl has 1 ring heteroatom selected from oxygen, sulfur, nitrogen, boron, silicon, and phosphorus.

[0078] Exemplary 3-membered heterocyclyl groups containing 1 heteroatom include, without limitation, aziridinyl, oxiranyl, thiorenyl. Exemplary 4-membered heterocyclyl groups containing 1 heteroatom include, without limitation, azetidinyl, oxetanyl and thietanyl. Exemplary 5-membered heterocyclyl groups containing 1 heteroatom include, without limitation, tetrahydrofuranyl, dihydrofuranyl, tetrahydrothiophenyl, dihydrothiophenyl, pyrrolidinyl, dihydropyrrolyl and pyrrolyl-2,5-dione. Exemplary 5- membered heterocyclyl groups containing 2 heteroatoms include, without limitation, dioxolanyl, oxathiolanyl and dithiolanyl. Exemplary 5-membered heterocyclyl groups containing 3 heteroatoms include, without limitation, triazolinyl, oxadiazolinyl, and thiadiazolinyl. Exemplary 6-membered heterocyclyl groups containing 1 heteroatom include, without limitation, piperidinyl, tetrahydropyranyl, dihydropyridinyl, and thianyl. Exemplary 6-membered heterocyclyl groups containing 2 heteroatoms include, without limitation, piperazinyl, morpholinyl, dithianyl, dioxanyl. Exemplary 6-membered heterocyclyl groups containing 2 heteroatoms include, without limitation, triazinanyl. Exemplary 7-membered heterocyclyl groups containing 1 heteroatom include, without limitation, azepanyl, oxepanyl and thiepanyl. Exemplary 8-membered heterocyclyl groupscontaining 1 heteroatom include, without limitation, azocanyl, oxecanyl and thiocanyl. Exemplary bicyclic heterocyclyl groups include, without limitation, indolinyl, isoindolinyl, dihydrobenzofuranyl, dihydrobenzothienyl, tetrahydrobenzothienyl, tetrahydrobenzofuranyl, tetrahydroindolyl, tetrahydroquinolinyl, tetrahydroisoquinolinyl, decahydroquinolinyl, decahydroisoquinolinyl, octahydrochromenyl, octahydroisochromenyl, decahydronaphthyridinyl, decahydro- 1,8-naphthyridinyl, octahydropyrrolo[3,2-b]pyrrole, indolinyl, phthalimidyl, naphthalimidyl, chromanyl, chromenyl, lH-benzo[e][l,4]diazepinyl, l,4,5,7-tetrahydropyrano[3,4-b] pyrrolyl, 5,6-dihydro-4H-furo[3,2-b]pyrrolyl, 6,7-dihydro-5H-furo[3,2-b]pyranyl, 5,7-dihydro-4H-thieno[2,3-c]pyranyl, 2,3-dihydro-lH-pyrrolo[2,3-b ]pyridinyl, 2,3-dihydrofuro[2,3-b]pyridinyl, 4,5,6,7-tetrahydro-lH-pyrrolo-[2,3-b]pyridinyl, 4,5,6,7-tetrahydrofuro[3,2-c]pyridinyl, 4,5,6,7-tetrahydrothieno [3,2- b] pyridinyl, 1, 2,3,4-tetrahydro-l,6-naphthyridinyl, and the like.

[0079] Heterocycloalkyl: The term "heterocycloalkyl," as used herein, is a non-aromatic ring wherein at least one atom is a heteroatom such as, but not limited to, nitrogen, oxygen, sulfur, or phosphorus, and the remaining atoms are carbon. The heterocycloalkyl group can be substituted or unsubstituted.

[0080] As understood from the above, alkyl, alkenyl, alkynyl, acyl, carbocyclyl, heterocyclyl, aryl, and heteroaryl groups, as defined herein, are, in certain embodiments, optionally substituted. Optionally substituted refers to a group which may be substituted or unsubstituted (e.g., "substituted" or "unsubstituted" alkyl, "substituted" or "unsubstituted" alkenyl, "substituted" or "unsubstituted" alkynyl, "substituted" or "unsubstituted" heteroalkyl, "substituted" or "unsubstituted" heteroalkenyl, "substituted" or 'unsubstituted" heteroalkynyl, "substituted" or "unsubstituted" carbocyclyl, "substituted" or "unsubstituted" heterocyclyl, "substituted" or "unsubstituted" aryl or "substituted" or "unsubstituted" heteroaryl group. In general, the term "substituted" means that at least one hydrogen present on a group is replaced with a permissible substituent, e.g., a substituent which upon substitution results in a stable compound, e.g., a compound which does not spontaneously undergo transformation such as by rearrangement, cyclization, elimination, or other reaction. Unless otherwise indicated, a "substituted" group has a substituent at one or more substitutable positions of the group, and when more than one position in any given structure is substituted, the substituent is either the same or different at each position. The term "substituted" is contemplated to include substitution with all permissible substituents of organic compounds, any of the substituents described herein that results in the formation of a stable compound. The present invention contemplates any and all such combinations in order to arrive at a stable compound. For purposes of this invention, heteroatoms such as nitrogen may have hydrogen substituents and / or any suitable substituent asdescribed herein which satisfy the valences of the heteroatoms and results in the formation of a stable moiety.

[0081] Exemplary carbon atom substituents include, but are not limited to, halogen, -CN, -NO2, -N3, -SO2, -SO3H, -OH, -ORaa, -ON(Rbb)2, -N(Rbb)2, -N(Rbb)3+X', -N(ORcc)Rbb, -SeH, -SeRaa, -SH, -SRaa, -SSRcc, -C(=O)Raa, -CO2H, -CHO, -C(ORcc)2, -CO2Raa, -OC(=O)Raa, -OCO2Raa, -C(=O)N(Rbb)2, -OC(=O)N(Rbb)2, -NRbbC(=O)Raa, -NRbbCO2Raa, -NRbbC(=O)N(Rbb)2, -C( = NRbb)Raa, -C(=NRbb)ORaa, -OC(=NRbb)Raa, - OC(=NRbb)ORaa, -C(=NRbb)N(Rbb)2, -OC(=NRbb)N(Rbb)2, -NRbbC(=NRbb)N(Rbb)2, -C(=O)NRbbSO2Raa, -NRbbSO2Raa, -SO2N(Rbb)2, -SO2Raa, -SO2ORaa, -OSO2Raa, -S(=O)Raa, -OS(=O)Raa, -Si(Raa)3-Osi(Raa)3-C(=S)N(Rbb)2, -C(=O)SRaa, -C(=S)SRaa, - SC(=S)SRaa, -SC(=O)SRaa, -OC(=O)SRaa, -SC(=O)ORaa, -SC(=O)Raa, -P(=O)2Raa, -OP(=O)2Raa, -P(=O)(Raa)2, -OP(=O)(Raa)2, -OP(=O)(ORcc)2, -P(=O)2N(Rbb)2, -OP(=O)2N(Rbb)2, - P(=O)(NRbb)2, -OP(=O)(NRbb)2, -NRbbP(=O)(ORcc)2, -NRbbP(=O)(NRbb)2, -P(RCC)2, - P(RCC)3, -OP(RCC)2, -OP(RCC)3, -B(Raa)2, -B(ORCC)2, -BRaa(ORcc), (C1-C50) alkyl, (C2-C50) alkenyl, (C2-C50) alkynyl, (C3-C14) carbocyclyl, 3-14 membered heterocyclyl, (Cs-C14) aryl, and 5-14 membered heteroaryl, wherein each alkyl, alkenyl, alkynyl, carbocyclyl, heterocyclyl, aryl, and heteroaryl is independently substituted with 0, 1, 2, 3, 4, or 5 Rddgroups;or two geminal hydrogens on a carbon atom are replaced with the group =0, =S, =NN(Rbb)2, =NNRbbC(=O)Raa, =NNRbbC(=O)ORaa, =NNRbbS(=O)2Raa, =NRbb, or =NORCC;

[0082] each instance of Raais, independently, selected from (C1-C50) alkyl, (C2-C50) alkenyl, (C2-C50) alkynyl, (C3-Cio) carbocyclyl, 3-14 membered heterocyclyl, (C6-C14) aryl, and 5-14 membered heteroaryl, or two Raagroups are joined to form a 3-14 membered heterocyclyl or 5-14 membered heteroaryl ring, wherein each alkyl, alkenyl, alkynyl, carbocyclyl, heterocyclyl, aryl, and heteroaryl is independently substituted with 0, 1, 2, 3, 4, or 5 Rddgroups;

[0083] each instance of Rbbis, independently, selected from hydrogen, -OH, -ORaa, - N(RCC)2, -CN, -C(=O)Raa, -C(=O)N(RCC)2, -CO2Raa, -SO2Raa, -C(=NRcc)ORaa, - C(=NRCC)N(RCC)2, -SO2N(RCC)2, -SO2RCC, -SO2ORCC, -SORaa, -C(=S)N(RCC)2, -C(=O)SRCC, - C(=S)SRCC, -P(=O)2Raa, -P(=O)(Raa)2, -P(=O)2N(RCC)2, -P(=O)(NRCC)2, (C1-C50) alkyl, (C2-C50) alkenyl, (C2-C50) alkynyl, (C3-C10) carbocyclyl, 3-14 membered heterocyclyl, (C3-C14) aryl, and 5-14 membered heteroaryl, or two Rbbgroups, together with the heteroatom to which they are attached, form a 3-14 membered heterocyclyl or 5-14 membered heteroaryl ring, wherein each alkyl, alkenyl, alkynyl, carbocyclyl, heterocyclyl, aryl, and heteroaryl is independently substituted with 0, 1, 2, 3, 4, or 5 Rddgroups;

[0084] each instance of Rccis, independently, selected from hydrogen, (C1-C50) alkyl, (C2-C50) alkenyl, (C2-C50) alkynyl, (C3-C10) carbocyclyl, 3-14 membered heterocyclyl, (C6-C14) aryl, and 5-14 membered heteroaryl, or two Rccgroups, together with the heteroatom to which they are attached, form a 3-14membered heterocyclyl or 5-14 membered heteroaryl ring, wherein each alkyl, alkenyl, alkynyl, carbocyclyl, heterocyclyl, aryl, and heteroaryl is independently substituted with 0, 1, 2, 3, 4, or 5 Rddgroups;

[0085] each instance of Rddis, independently, selected from halogen, -CN, -NO2, -N3, -SO2H, -SO3H, -OH, -ORee, -ON(Rff)2, -N(Rff)2, -N(Rff)3+X', -N(ORee)Rff, -SH, -SRee, - SSRee, -C(=O)Ree, -CO2H, -CO2Ree, -OC(=O)Ree, -OCO2Ree, -C(=O)N(Rff)2, - OC(=O)N(Rff)2, -NRffC(=O)Ree, -NRffCO2Ree, -NRffC(=O)N(Rff)2, -C(=NRff)ORee, - OC(=NRff)Ree, -OC(=NRff)ORee, -C(=NRff)N(Rff)2, -OC(=NRff)N(Rff)2, -NRffC( = NRff)N(Rff)2, -NRffSO2Ree, -SO2N(Rff)2, -SO2Ree, -SO2ORee, -OSO2Ree, -S(=O)Ree, -Si(Ree)3, -Osi(Ree)3, -C(=S)N(Rff)2, -C(=O)SRee, -C(=S)SRee, -SC(=S)SRee, -P(=O)2Ree, - P(=O)(Ree)2, -OP(=O)(Ree)2, -OP(=O)(ORee)2, (C1-C50) alkyl, (C2-C50) alkenyl, (C2-C50) alkynyl, (C3-C10) carbocyclyl, 3-10 membered heterocyclyl, (C5-C10) aryl, 5-10 membered heteroaryl, wherein each alkyl, alkenyl, alkynyl, carbocyclyl, heterocyclyl, aryl, and heteroaryl is independently substituted with 0, 1, 2, 3, 4, or 5 Rgggroups, or two geminal Rddsubstituents can be joined to form =0 or =S;

[0086] each instance of Reeis, independently, selected from (C1-C50) alkyl, (C2-C50) alkenyl, (C2-C50) alkynyl, (C3-C10) carbocyclyl, (C5-C10) aryl, 3-10 membered heterocyclyl, and 3-10 membered heteroaryl, wherein each alkyl, alkenyl, alkynyl, carbocyclyl, heterocyclyl, aryl, and heteroaryl is independently substituted with 0, 1, 2, 3, 4, or 5 Rgggroups;

[0087] each instance of Rffis, independently, selected from hydrogen, (C1-C50) alkyl, (C2-C50) alkenyl, (C2-C50) alkynyl, (C3-C10) carbocyclyl, 3-10 membered heterocyclyl, (C5-C10) aryl and 5-10 membered heteroaryl, or two Rffgroups, together with the heteroatom to which they are attached, form a 3-14 membered heterocyclyl or 5-14 membered heteroaryl ring, wherein each alkyl, alkenyl, alkynyl, carbocyclyl, heterocyclyl, aryl, and heteroaryl is independently substituted with 0, 1, 2, 3, 4, or 5 Rgggroups; and

[0088] each instance of Rggis, independently, halogen, -CN, -NO2, -N3, -SO2H, -SO3H, -OH, -O(Ci-C50) alkyl, -ON((CI-C50) alkyl)2, -N((CI-C50) alkyl)2, -N((CI-C50) alkyl)3+X', -NH((Ci-C50)alkyl)2+X-, -NH2((CI-C5O) alkyl) +X’, -NH3+X -N(O(CI-C50) alkyl)((Ci-C50) alkyl), -N(OH)((CI-C50) alkyl), -NH(OH), -SH, -S(Ci-C50) alkyl, -SS((Ci-C50) alkyl), -C(=O)((Ci-C50) alkyl), -CO2H, -CO2((Ci-C50) alkyl), -OC(=O)((Ci-C50) alkyl), -OCO2((C1-C50) alkyl), -C(=O)NH2, -C(=O)N((CI-C50) alkyl)2, -OC(=O)NH((CI-C50) alkyl), -NHC(=O)((C1-C50) alkyl), -N((CI-C5O) alkyl)C(=0)((Ci-C5o) alkyl), -NHCO2((C1-C50) alkyl), -NHC(=O)N((CI-C50) alkyl)2, -NHC(=O)NH((CI-C50) alkyl), -NHC(=O)NH2, -C(=NH)O((CI-C50) alkyl), -OC(=NH)((CI-C50) alkyl), -OC(=NH)O(CI-C50) alkyl, - C(=NH)N((CI-C50) alkyl)2, -C(=NH)NH((CI-C50) alkyl), -C(=NH)NH2, -OC(=NH)N((Ci-C5o)alkyl)2, -OC(NH)NH((CI-C50) alkyl), -OC(NH)NH2, -NHC(NH)N((CI-C50) alkyl)2, -NHC(=NH)NH2, -NHSO2((C1-C50) alkyl), -SO2N((C1-C50) alkyl)2, -SO2NH((C1-C50) alkyl), -SO2NH2, -SO2((C1-C50) alkyl), -SO2O((C1-C50) alkyl), -OSO2((Ci-C6) alkyl), -SO((Ci-C6) alkyl), -Si((Ci-C50) alkyl)3, --OSi((C1-C6) alkyl)3, -C(=S)N((CI-C5O) alkyl)2, -C(=S)NH((C1-C50) alkyl), -C(=S)NH2, -C(=O)S((Ci-C6) alkyl), -C(=S)S((C1-C6) alkyl), -SC(=S)S((Ci-C6) alkyl), -P(=O)2((C1-C50) alkyl), -P(=O)((C1-C50) alkyl)2, -OP(=O)((C1-C50) alkyl)2, -OP(=O)(O(C1-C50) alkyl)2, (C1-C50) alkyl, (C2-C50) alkenyl, (C2-C50) alkynyl, (C3-C10) carbocyclyl, (Cs-Cio) aryl, 3-10 membered heterocyclyl, 5-10 membered heteroaryl; or two geminal Rggsubstituents can be joined to form =0 or =S; wherein X" is a counterion.

[0089] As used herein, the term "halo" or "halogen" refers to fluorine (fluoro, -F), chlorine (chloro, -Cl), bromine (bromo, -Br), or iodine (iodo, -I).

[0090] As used herein, a "counterion" is a negatively charged group associated with a positively charged quaternary amine in order to maintain electronic neutrality. Exemplary counterions include halide ions (e.g., F⁻, Cl⁻, Br⁻, I⁻), NO3-, ClO4-, OH-, H2PO4-, HSO4-, sulfonate ions (e.g., methanesulfonate, trifluoromethanesulfonate, p-toluenesulfonate, benzenesulfonate, 10-camphor sulfonate, naphthalene-2-sulfonate, naphthalene-l-sulfonic acid-5-sulfonate, ethan-l-sulfonic acid-2-sulfonate, and the like), and carboxylate ions (e.g., acetate, ethanoate, propanoate, benzoate, glycerate, lactate, tartrate, glycolate, and the like).

[0091] Nitrogen atoms can be substituted or unsubstituted as valency permits, and include primary, secondary, tertiary, and quaternary nitrogen atoms. Exemplary nitrogen atom substituents include, but are not limited to, hydrogen, -OH, -ORaa, -N(RCC)2, -CN, - C(=O)Raa, -C(=O)N(RCC)2, -CO2Raa, -SO2Raa, -C(=NRbb)Raa, -C(=NRcc)ORaa, -C(=NRcc)N(Rcc)2, -SO2N(Rcc)2, -SO2Rcc, -SO2ORcc, -SORaa, -C(=S)N(Rcc)2, -C(=O)SRcc, -C(=S)SRcc, -P(=O)2Raa, -P(=O)(Raa)2, -P(=O)2N(Rcc)2, -P(=O)(NRcc)2, (C1-C50) alkyl, (C2-C50) alkenyl, (C2-C50) alkynyl, (C3-C10) carbocyclyl, 3-14 membered heterocyclyl, (C6-C14) aryl, and 5-14 membered heteroaryl, or two Rccgroups, together with the N atom to which they are attached, form a 3-14 membered heterocyclyl or 5-14 membered heteroaryl ring, wherein each alkyl, alkenyl, alkynyl, carbocyclyl, heterocyclyl, aryl, and heteroaryl is independently substituted with 0, 1, 2, 3, 4, or 5 Rddgroups, and wherein Raa, Rbb, Rccand Rddare as defined above.

[0092] In certain embodiments, the substituent present on a nitrogen atom is a nitrogen protecting group (also referred to as an amino protecting group). Nitrogen protecting groups are well known in the art and include those described in detail in Protecting Groups in Organic Synthesis, T. W. Greene and P. G. M. Wuts, 3rd edition, John Wiley & Sons, 1999, incorporated herein by reference.

[0093] For example, nitrogen protecting groups such as amide groups (e.g., - C(=O)Raa) include, but are not limited to, formamide, acetamide, chloroacetamide, trichloroacetamide, trifluoroacetamide, phenylacetamide, 3-phenylpropanamide, picolinamide, 3-pyridylcarboxamide, N-benzoylphenylalanyl derivative, benzamide, p-phenylbenzamide, o-nitophenylacetamide, o-nitrophenoxyacetamide, acetoacetamide, (N'-dithiobenzyloxyacylamino)acetamide, 3-(p-hydroxyphenyl)propanamide, 3-(o-nitrophenyl)propanamide, 2-methyl-2-(o-nitrophenoxy)propanamide, 2-methyl-2-(o-phenylazophenoxy)propanamide, 4-chlorobutanamide, 3-methyl-3-nitrobutanamide, o-nitrocinnamide, N-acetylmethionine derivative, o-nitrobenzamide and o-(benzoyloxymethyl)benzamide.

[0094] Nitrogen protecting groups such as carbamate groups (e.g., -C(=O)ORaa) include, but are not limited to, methyl carbamate, ethyl carbamate, 9-fluorenylmethyl carbamate (Fmoc), 9-(2-sulfo)fluorenylmethyl carbamate, 9-(2,7-dibromo)fluoroenylmethyl carbamate, 2,7-di-t-butyl-[9-(10,10-dioxo-10,10,10,10-tetrahydrothioxanthyl)]methyl carbamate (DBD-Tmoc), 4-methoxyphenacyl carbamate (Phenoc), 2,2,2-trichloroethyl carbamate (Troc), 2-trimethylsilylethyl carbamate (Teoc), 2-phenylethyl carbamate (hZ), 1-(1-adamantyl)-1-methylethyl carbamate (Adpoc), 1,1-dimethyl-2-haloethyl carbamate, 1,1-dimethyl-2,2-dibromoethyl carbamate (DB-t-BOC), 1,1-dimethyl-2,2,2-trichloroethyl carbamate (TCBOC), 1-methyl-1-(4-biphenylyl)ethyl carbamate (Bpoc), 1-(3,5-di-t-butylphenyl)-1-methylethyl carbamate (t-Bumeoc), 2-(2'-and 4'-pyridyl)ethyl carbamate (Pyoc), 2-(N, N-dicyclohexylcarboxamido)ethyl carbamate, t-butyl carbamate (BOC), 1-adamantyl carbamate (Adoc), vinyl carbamate (Voc), allyl carbamate (Alloc), 1-isopropylal lyl carbamate (Ipaoc), cinnamyl carbamate (Coc), 4-nitrocinnamyl carbamate (Noc), 8-quinolyl carbamate, N-hydroxypiperidinyl carbamate, alkyldithio carbamate, benzyl carbamate (Cbz), p-methoxybenzyl carbamate (Moz), p-nitobenzyl carbamate, p-bromobenzyl carbamate, p-chlorobenzyl carbamate, 2.4-dichlorobenzyl carbamate, 4-methylsulfinylbenzyl carbamate (Msz), 9-anthrylmethyl carbamate, diphenylmethyl carbamate, 2-methylthioethyl carbamate, 2-methylsulfonylethyl carbamate, 2-(p-toluenesulfonyl)ethyl carbamate, [2-(1,3-dithianyl)]methyl carbamate (Dmoc), 4- methylthiophenyl carbamate (Mtpc), 2,4-dimethylthiophenyl carbamate (Bmpc), 2-phosphonioethyl carbamate (Peoc), 2-triphenylphosphonioisopropyl carbamate (Ppoc), 1,1-dimethyl-2-cyanoethyl carbamate, m-chloro-p-acyloxybenzyl carbamate, p-(dihydroxyboryl)benzyl carbamate, 5-benzisoxazolylmethyl carbamate, 2-(trifluoromethyl)-6-chromonylmethyl carbamate (Tcroc), m-nitrophenyl carbamate, 3.5-dimethoxybenzyl carbamate, o-nitrobenzyl carbamate, 3,4-dimethoxy-6-nitrobenzyl carbamate, phenyl(o-nitrophenyl)methyl carbamate, t-amyl carbamate, S-benzyl thiocarbamate, p-cyanobenzyl carbamate, cyclobutyl carbamate, cyclohexyl carbamate, cyclopentyl carbamate, cyclopropylmethyl carbamate, p-decyloxybenzyl carbamate, 2,2-dimethoxyacylvinyl carbamate, o-(N, N-dimethylcarboxamido)benzyl carbamate, l,l-dimethyl-3-(N, N-dimethylcarboxamido)propyl carbamate, 1,1-dimethylpropynyl carbamate, di(2-pyridyl)methyl carbamate, 2-furanylmethyl carbamate, 2-iodoethyl carbamate, isoborynl carbamate, isobutyl carbamate, isonicotinyl carbamate, p-(p'-methoxyphenylazo)benzyl carbamate, 1-methylcyclobutyl carbamate, 1-methylcyclohexyl carbamate, 1-methyl-l-cyclopropylmethyl carbamate, 1-methyl-1-(3,5-dimethoxyphenyl)ethyl carbamate, 1-methyl-1-(p-phenylazophenyl)ethyl carbamate, 1-methyl-1-phenylethyl carbamate, 1- methyl-l-(4-pyridyl)ethyl carbamate, phenyl carbamate, p-(phenylazo)benzyl carbamate, 2,4,6-tri-t-butylphenyl carbamate, 4-(trimethylammonium)benzyl carbamate, and 2,4,6-trimethylbenzyl carbamate.

[0095] Nitrogen protecting groups such as sulfonamide groups (e.g., -S(=O)2Raa) include, but are not limited to, p-toluenesulfonamide (Ts), benzenesulfonamide, 2,3,6,-trimethyl-4-methoxybenzenesulfonamide (Mtr), 2,4,6-trimethoxybenzenesulfonamide (Mtb), 2,6-dimethyl-4-methoxybenzenesulfonamide (Pme), 2,3,5,6-tetramethyl-4-methoxybenzenesulfonamide (Mte), 4-methoxybenzenesulfonamide (Mbs), 2,4,6- trimethylbenzenesulfonamide (Mts), 2,6-dimethoxy-4-methylbenzenesulfonamide (iMds), 2,2,5,7,8-pentamethylchrornan-6-sulfonamide (Pmc), methanesulfonamide (Ms), p-trimethylsilylethanesulfonamide (SES), 9-anthracenesulfonamide, 4-(4',8'-dimethoxynaphthylmethyl)benzenesulfonamide (DNMBS), benzylsulfonamide, trifluoromethylsulfonamide, and phenacylsulfonamide.

[0096] Other nitrogen protecting groups include, but are not limited to, phenothiazinyl-(10)-acyl derivative, N'-p-toluenesulfonylaminoacyl derivative, N' -phenylaminothioacyl derivative, N-benzoylphenylalanyl derivative, N-acetylmethionine derivative, 4,5-diphenyl-3-oxazolin-2-one, N-phthalimide, N-dithiasuccinimide (Dts), N-2,3-diphenylmaleimide, N-2,5-dimethylpyrrole, N-1,1,4,4-tetramethyldisilylazacyclopentane adduct (STABASE), 5-substituted 1,3-dimethyl-1,3,5-triazacyclohexan-2-one, 5-substituted 1,3-dibenzyl-1,3,5-triazacyclohexan-2-one, 1-substituted 3,5-dinitro-4-pyridone, N-methylamine, N-allylamine, N-[2- (trimethylsilyl)ethoxy]methylamine (SEM), N-3-acetoxypropylamine, N-(1-isopropyl-4-nitro-2-oxo-3-pyrrolin-3-yl)amine, quaternary ammonium salts, N-benzylamine, N-di(4-methoxyphenyl)methylamine, N-5-dibenzosuberylamine, N-triphenylmethylamine (Tr), N-[(4-methoxyphenyl)diphenylmethyl]amine (MMTr), N-9-phenylfluorenylamine (PhF), N-2,7-dichloro-9-fluorenylmethyleneamine, N-ferrocenylmethylamino (Fem), N-2- picolylamino N'-oxide, N-l,l-dimethylthiomethyleneamine, N-benzylideneamine, N-p-methoxybenzylideneamine, N-diphenylmethyleneamine, N-[(2-pyridyl)mesityl]methyleneamine, N-(N', N'-dimethylaminomethylene)amine, N, N' -isopropylidenediamine, N-p-nitrobenzylideneamine, N-salicylideneamine, N-5- chlorosalicylideneamine, N-(5-chloro-2-hydroxyphenyl) phenylmethyleneamine, N-cyclohexylideneamine, N-(5,5-dimethyl-3-oxo-l-cyclohexenyl)amine, N-borane derivative, N-diphenylborinic acid derivative, N-[phenyl(pentaacylchromium- or tungsten)acyl]amine, N-copper chelate, N-zinc chelate, N-nitroamine, N-nitrosoamine, amine N-oxide, diphenylphosphinamide (Dpp), dimethylthiophosphinamide (Mpt), diphenylthiophosphinamide (Ppt), dialkyl phosphoramidates, dibenzyl phosphoramidate, diphenyl phosphoramidate, benzenesulfenamide, o-nitrobenzenesulfenamide (Nps), 2,4-dinitrobenzenesulfenamide, pentachlorobenzenesulfenamide, 2-nitro-4-methoxybenzenesulfenamide, triphenylmethylsulfenamide, and 3-nitropyridinesulfenamide (Npys).

[0097] In certain embodiments, the substituent present on an oxygen atom is an oxygen protecting group (also referred to as a hydroxyl protecting group). Oxygen protecting groups are well known in the art and include those described in detail in Protecting Groups in Organic Synthesis, T. W. Greene and P. G. M. Wuts, 3rd edition, John Wiley & Sons, 1999, incorporated herein by reference.

[0098] Exemplary oxygen protecting groups include, but are not limited to, methyl, methoxylmethyl (MOM), methylthiomethyl (MTM), t-butylthiomethyl, (phenyldimethylsilyl)methoxymethyl (SMOM), benzyloxymethyl (BOM), p-methoxybenzyloxymethyl (PMBM), (4-methoxyphenoxy)methyl (p-AOM), guaiacolmethyl (GUM), t-butoxymethyl, 4-pentenyloxymethyl (POM), siloxymethyl, 2-methoxyethoxymethyl (MEM), 2,2,2-trichloroethoxymethyl, bis(2-chloroethoxy)methyl, 2-(trimethylsilyl)ethoxymethyl (SEMOR), tetrahydropyranyl (THP), 3-bromotetrahydropyranyl, tetrahydrothiopyranyl, 1-methoxycyclohexyl, 4- methoxytetrahydropyranyl (MTHP), 4-methoxytetrahydrothiopyranyl, 4- methoxytetrahydrothiopyranyl S, S-dioxide, l-[(2-chloro-4-methyl)phenyl]-4-methoxypiperidin-4-yl (CTMP), l,4-dioxan-2-yl, tetrahydrofuranyl, tetrahydrothiofuranyl, 2,3,3a,4,5,6,7,7a-octahydro-7,8,8-trimethyl-4,7-methanobenzofuran-2-yl, 1-ethoxyethyl, l-(2-chloroethoxy)ethyl, 1-methyl-l-methoxyethyl, 1-methyl-l-benzyloxyethyl, 1-methyl-l-benzyloxy-2-fluoroethyl, 2,2,2-trichloroethyl, 2-trimethylsilylethyl, 2- (phenylselenyl)ethyl, t-butyl, allyl, p-chlorophenyl, p-methoxyphenyl, 2,4-dinitrophenyl, benzyl (Bn), p-methoxybenzyl, 3,4-dimethoxybenzyl, o-nitrobenzyl, p-nitrobenzyl, p-halobenzyl, 2,6-dichlorobenzyl, p-cyanobenzyl, p-phenylbenzyl, 2-picolyl, 4-picolyl, 3- methyl-2-picolyl N-oxido, diphenylmethyl, p,p'-dinitrobenzhydryl, 5-dibenzosuberyl, triphenylmethyl, a-naphthyldiphenylmethyl, p-methoxyphenyldiphenylmethyl, di(p-methoxyphenyl)phenylmethyl, tri(p-methoxyphenyl)methyl, 4-(4'-bromophenacyloxyphenyl)diphenylmethyl, 4,4',4"-tris(4,5-dichlorophthalimidophenyl)methyl, 4,4',4"-tris(levulinoyloxyphenyl)methyl, 4,4',4"-tris(benzoyloxyphenyl)methyl, 3-(imidazol-l-yl)bis(4',4"-dimethoxyphenyl)methyl, l,l-bis(4-methoxyphenyl)-l'-pyrenylmethyl, 9-anthryl, 9-(9-phenyl)xanthenyl, 9-(9-phenyl-10-oxo)anthryl, l,3-benzodisulfuran-2-yl, benzisothiazolyl S, S-dioxido, trimethylsilyl (TMS), triethylsilyl (TES), triisopropylsilyl (TIPS), dimethylisopropylsilyl (IPDMS), diethylisopropylsilyl (DEIPS), dimethylthexylsilyl, t-butyldimethylsilyl (TBDMS), t-butyldiphenylsilyl (TBDPS), tribenzylsilyl, tri-p-xylylsilyl, triphenylsilyl, diphenylmethylsilyl (DPMS), t-butylmethoxyphenylsilyl (TBMPS), formate, benzoylformate, acetate, chloroacetate, dichloroacetate, trichloroacetate, trifluoroacetate, methoxyacetate, triphenylmethoxyacetate, phenoxyacetate, p-chlorophenoxyacetate, 3- phenylpropionate, 4-oxopentanoate (levulinate), 4,4-(ethylenedithio)pentanoate (levulinoyldithioacetal), pivaloate, adamantoate, crotonate, 4-methoxycrotonate, benzoate, p-phenylbenzoate, 2,4,6-trimethylbenzoate (mesitoate), alkyl methyl carbonate, 9- fluorenylmethyl carbonate (Fmoc), alkyl ethyl carbonate, alkyl 2,2,2-trichloroethyl carbonate (Troc), 2-(trimethylsilyl)ethyl carbonate (TMSEC), 2-(phenylsulfonyl) ethyl carbonate (Psec), 2-(triphenylphosphonio) ethyl carbonate (Peoc), alkyl isobutyl carbonate, alkyl vinyl carbonate alkyl allyl carbonate, alkyl p-nitrophenyl carbonate, alkyl benzyl carbonate, alkyl p-methoxybenzyl carbonate, alkyl 3,4-dimethoxybenzyl carbonate, alkyl o-nitrobenzyl carbonate, alkyl p-nitrobenzyl carbonate, alkyl S-benzyl thiocarbonate, 4-ethoxy-l-napththyl carbonate, methyl dithiocarbonate, 2-iodobenzoate, 4-azidobutyrate, 4-nitro-4-methylpentanoate, o-(dibromomethyl)benzoate, 2-formylbenzenesulfonate, 2-(methylthiomethoxy)ethyl, 4-(methylthiomethoxy)butyrate, 2- (methylthiomethoxymethyl)benzoate, 2,6-dichloro-4-methylphenoxyacetate, 2,6-dichloro-4-(l,l,3,3-tetramethylbutyl)phenoxyacetate, 2,4-bis(l,l-dimethylpropyl)phenoxyacetate, chlorodiphenylacetate, isobutyrate, monosuccinoate, (E)-2-methyl-2-butenoate, o-(methoxyacyl)benzoate, a-naphthoate, nitrate, alkyl N, N, N', N'-tetramethylphosphorodiamidate, alkyl N-phenylcarbamate, borate, dimethylphosphinothioyl, alkyl 2,4-dinitrophenylsulfenate, sulfate, methanesulfonate (mesylate), benzylsulfonate, and tosylate (Ts).

[0099] In certain embodiments, the substituent present on a sulfur atom is a sulfur protecting group (also referred to as a thiol protecting group). Sulfur protecting groups are well known in the art and include those described in detail in Protecting Groups in Organic Synthesis, T. W. Greene and P. G. M. Wuts, 3rd edition, John Wiley & Sons, 1999, incorporated herein by reference.

[0100] Exemplary sulfur protecting groups include, but are not limited to, alkyl, benzyl, p-methoxybenzyl, 2,4,6-trimethylbenzyl, 2,4,6-trimethoxybenzyl, o-hydroxybenzyl, p-hydroxybenzyl, o-acetoxybenzyl, p-acetoxybenzyl, p-nitrobenzyl, 4-picolyl, 2-quinolinylmethyl, 2-picolyl N-oxido, 9-anthrylmethyl, 9-fluorenylmethyl, xanthenyl, ferrocenylmethyl, diphenylmethyl, bis(4-methoxyphenyl)methyl, 5-dibenzosuberyl, triphenylmethyl, diphenyl-4-pyridylmethyl, phenyl, 2,4-dinitrophenyl, t-butyl, 1-adamantyl, methoxymethyl (MOM), isobutoxymethyl, benzyloxymethyl, 2-tetrahydropyranyl, benzylthiomethyl, phenylthiomethyl, thiazolidino, acetamidomethyl, trimethylacetamidomethyl, benzamidomethyl, allyloxycarbonylaminomethyl, phenylacetamidomethyl, phthalimidomethyl, acetylmethyl, carboxymethyl, cyanomethyl, (2-nitro-l-phenyl)ethyl, 2-(2,4-dinitrophenyl)ethyl, 2-cyanoethyl, 2-(Trimethylsilyl)ethyl, 2,2-bis(carboethoxy)ethyl, (1-m-nitrophenyl-2-benzoyl)ethyl, 2-phenylsulfonylethyl, 2-(4-methylphenylsulfonyl)-2-methylprop-2-yl, acetyl, benzoyl, trifluoroacetyl, N-[[(p-biphenylyl)isopropoxy]carbonyl]-N-methyl]- y-aminothiobutyrate, 2,2,2-trichloroethoxycarbonyl, t-butoxycarbonyl, benzyloxycarbonyl, p-methoxybenzyloxycarbonyl, N-ethyl, N-methoxymethyl, sulfonate, sulfenylthiocarbonate, 3-nitro-2-pyridinesulfenyl sulfide, oxathiolone.Compounds of the Invention

[0101] Liposomal-based vehicles are considered as an attractive carrier for therapeutic agents and remain subject to continued development efforts. While liposomal-based vehicles that comprise certain lipid components have shown promising results with regard to encapsulation, stability and site localization, there remains a great need for improvement of liposomal-based delivery systems. For example, a significant drawback of liposomal delivery systems relates to the construction of liposomes that have sufficient stability to be manufactured, stored, and to reach desired target cells and / or intracellular compartments, and the ability of such liposomal delivery systems to efficiently release their encapsulated materials to such target cells.

[0102] In particular, there remains a need for cationic lipids that are effective for intramuscular and / or intravenous delivery of mRNA (e.g., for preventing or treating Flu, acne, chlamydia, or a disease or disorder induced by infection with P. gingivalis). There also remains a need for improved lipids compounds that demonstrate improved pharmacokinetic properties, and which are capable of delivering macromolecules, such as nucleic acids, to a wide variety cell types and tissues with enhanced efficiency. Importantly, there also remains a particular need for novel lipid compounds that are characterized as having improved safety profiles and are capable of efficiently delivering encapsulated nucleic acids and polynucleotides to targeted cells, tissues and organs.

[0103] Described herein is a novel class of cationic lipid compounds for improved in vivo delivery of therapeutic agents, such as nucleic acids (e.g., for treating or preventing Flu, acne, chlamydia, or a disease or disorder induced by infection with P. gingivalis). In particular, a cationic lipid described herein may be used, optionally with other lipids, to formulate a lipid-based nanoparticle (e.g., liposome) for encapsulating therapeutic agents, such as nucleic acids (e.g., DNA, siRNA, mRNA, microRNA) for therapeutic use, such as disease treatment and prevention (e.g for use as a vaccine e.g., for Flu, or for the prevention or treatment of acne, chlamydia, or a disease or disorder induced by infection with P. gingivalis) purposes.

[0104] In embodiments, compounds of the invention as described herein can provide one or more desired characteristics or properties. That is, in certain embodiments, compounds of the invention as described herein can be characterized as having one or more properties that afford such compounds advantages relative to other similarly classified lipids. For example, compounds disclosed herein can allow for the control and tailoring of the properties of liposomal compositions (e.g., lipid nanoparticles) of which they are a component. In particular, compounds disclosed herein can be characterized by enhanced transfection efficiencies and their ability to provoke specific biological outcomes. Such outcomes can include, for example enhanced cellular uptake,endosomal / lysosomal disruption capabilities and / or promoting the release of encapsulated materials (e.g., polynucleotides) intracellularly. The compounds disclosed herein can also be characterized by achieving high levels of peptide or protein expression when delivering mRNA encoding for said peptide or protein by intravenous, intrathecal, intramuscular, intranasal, sublingual, or by pulmonary delivery, optionally through nebulization. The compounds disclosed herein can also be characterized by achieving high Hemagglutination Inhibition (HAI) when delivering mRNA encoding an influenza antigen by intramuscular delivery. Additionally, the compounds disclosed herein have advantageous pharmacokinetic properties, biodistribution, and efficiency.

[0105] Additionally, the cationic lipids of the present invention have cleavable groups such as ester groups. These cleavable groups (e.g. esters, amides, disulphides, carbonates, carbamates, carbamides, and thiocarbamates) are contemplated to improve biodegradability and thus contribute to the lipids' favorable safety profiles.

[0106] It is contemplated that the cationic lipids of the present invention are capable of highly effective in vivo intramuscular and / or intravenous delivery of therapeutic agents and vaccines (e.g., for treating or preventing Flu, acne, chlamydia, or a disease or disorder induced by infection with P. gingivalis). It is also contemplated that lipid nanoparticles comprising the cationic lipids of the present invention are capable of highly effective in vivo delivery while maintaining a favorable safety profile. It is also contemplated that lipid nanoparticles comprising the cationic lipids of the present invention may exhibit degradation in vivo.

[0107] Provided herein are compounds which are cationic lipids. In embodiments, the cationic lipids include compounds having a structure according to Formula (I):(I) or a pharmaceutically acceptable salt thereof, wherein:, -0-, -S-, and -(C1-C6alkylene)-, wherein the left hand side of each depicted structure is bound to the -(CH2)a-;Z1is selected fromthe right hand side of each depicted structure is bound to the -(CH2)a-;each R is independently selected from:, wherein each R1is independently selected from optionally substituted alkyl, optionally substituted alkenyl, and optionally substituted alkynyl;, wherein each R2is independently selected from optionally substituted alkyl, optionally substituted alkenyl, optionally substituted alkynyl, and -W1-X1, wherein each W1is independently selected from optionally substituted alkylene and optionally substituted alkenylene, andeach X1is independently selected from -*O-(C=O)-optionally substituted alkyl, -(*C=O)-O- optionally substituted alkyl, -*O-(C=O)-optionally substituted alkenyl, and -(*C=O)-O-optionally substituted alkenyl, wherein the atom marked with a * is connected to W1;(iii), wherein each R3is independently selected from optionally substituted alkyl, optionally substituted alkenyl, and optionally substituted alkynyl; and(iv), wherein each R4is independently selected from optionally substituted alkyl, optionally substituted alkenyl, and optionally substituted alkynyl; (v) optionally substituted alkyl or optionally substituted alkenyl;each R5is selected from hydrogen and optionally substituted (Ci-Cs) alkyl;each R6is selected from optionally substituted (Ci-Cs) alkyl;each a is independently selected from 1, 2, 3, 4, 5, 6, 7, 8, 9 and 10;each b is 1, 2, 3, 4 or 5;each c is 1, 2, 3, 4, or 5;each d is independently selected from 2, 3, 4, 5, 6, 7, 8, 9 and 10;each e is independently selected from 2, 3, 4, 5, 6, 7, 8, 9 and 10; andeach f is independently selected from 2, 3, 4, 5, 6, 7, 8, 9 and 10.

[0108] In embodiments, the cationic lipids include compounds having a structure according to Formula (IA):(IA) or a pharmaceutically acceptable salt thereof.

[0109] In embodiments, the cationic lipids include compounds having a structure according to Formula (IB):(IB) or a pharmaceutically acceptable salt thereof.

[0110] In embodiments, the cationic lipids include compounds having a structure according to Formula (IC):(IC) or a pharmaceutically acceptable salt thereof, wherein each R2A, R2B, R2Cand R2Dis independently selected from optionally substituted alkyl, optionally substituted alkenyl, optionally substituted alkynyl, and -W1-X1,wherein each W1is independently selected from optionally substituted alkylene and optionally substituted alkenylene, andeach X1is independently selected from -*O-(C=O)-optionally substituted alkyl, -(*C=O)-O- optionally substituted alkyl, -*O-(C=O)-optionally substituted alkenyl, and -(*C=O)-O-optionally substituted alkenyl, wherein the atom marked with a * is connected to W1.

[0111] In embodiments, the cationic lipids include compounds having a structure according to Formula (ID):(ID) or a pharmaceutically acceptable salt thereof, wherein each R2A, R2B, R2Cand R2Dis independently selected from optionally substituted alkyl, optionally substituted alkenyl, optionally substituted alkynyl, and -W1-X1,wherein each W1is independently selected from optionally substituted alkylene and optionally substituted alkenylene, andeach X1is independently selected from -*O-(C=O)-optionally substituted alkyl, -(*C=O)-O- optionally substituted alkyl, -*O-(C=O)-optionally substituted alkenyl, and -(*C=O)-O-optionally substituted alkenyl, wherein the atom marked with a * is connected to W1.

[0112] In embodiments, each a is 1. In embodiments, each a is 2. In embodiments, each a is 3. In embodiments, each a is 4. In embodiments, each a is 5. In embodiments, each a is 6. In embodiments, each a is 7. In embodiments, each a is 8. In embodiments, each a is 9. In embodiments, each a is 10.

[0113] In embodiments, each a is independently selected from 2, 3 and 4. In embodiments, each a is 2. In embodiments, each a is 3. In embodiments, each a is 4.

[0114] In embodiments, the value for the a on the left hand side of the depicted Formula is 2 and the value for the a on the right hand side of the depicted Formula is 4. In embodiments, the value for the a on the left hand side of the depicted Formula is 3 and the value for the a on the right hand side of the depicted Formula is 4. In embodiments, the value for the a on the left hand side of the depicted Formula is 4 and the value for the a on the right hand side of the depicted Formula is 3. In embodiments, the value for the a on the left hand side of the depicted Formula is 4 and the value for the a on the right hand side of the depicted Formula is 4.

[0115] In embodiments, each a is the same. In embodiments, each a is different.

[0116] In embodiments, each b is 1. In embodiments, each b is 2. In embodiments, each b is 3. In embodiments, each b is 4. In embodiments, each b is 5.

[0117] In embodiments, b is selected from 2, 3 and 4. In embodiments, b is selected from 2 and 4. In embodiments, b is 2. In embodiments, b is 3. In embodiments, b is 4.

[0118] In embodiments, each c is 1. In embodiments, each c is 2. In embodiments, each c is 3. In embodiments, each c is 4. In embodiments, each c is 5.

[0119] In embodiments, c is selected from 2, 3 and 4. In embodiments, c is 2. In embodiments, c is 3. In embodiments, c is 4.

[0120] In embodiments, each d is 2. In embodiments, each d is 3. In embodiments, each d is 4. In embodiments, each d is 5. In embodiments, each d is 6. In embodiments, each d is 7. In embodiments, each d is 8. In embodiments, each d is 9. In embodiments, each d is 10.

[0121] In embodiments, each d is independently selected from 5, 6 and 7. In embodiments, each d is independently selected from 5 and 7. In embodiments, d is 5. In embodiments, d is 6. In embodiments, d is 7.

[0122] In embodiments, each e is 2. In embodiments, each e is 3. In embodiments, each e is 4. In embodiments, each e is 5. In embodiments, each e is 6. In embodiments, each e is 7. In embodiments, each e is 8. In embodiments, each e is 9. In embodiments, each e is 10.

[0123] In embodiments, e is 6.

[0124] In embodiments, each f is 2. In embodiments, each f is 3. In embodiments, each f is 4. In embodiments, each f is 5. In embodiments, each f is 6. In embodiments, each f is 7. In embodiments, each f is 8. In embodiments, each f is 9. In embodiments, each f is 10.

[0125] In embodiments, each f is independently selected from 3 and 4.

[0126] In embodiments, A1is a bond. In embodiments, A1is wherein the left hand side of each depicted structure is bound to the -(CHz)a-. In embodiments, A1isHwherein the left hand side of each depicted structure is bound to the -(CH2)a-. In embodiments, A1is wherein the left hand side of eachdepicted structure is bound to the -(CH2)a-. In embodiments,A1is wherein the left hand side of each depicted structure is bound to the -(CH2)a-. In embodiments, A1iswherein the left hand side of each depicted structure is bound to the -(CHz)a-. In embodiments, A1iswherein the left hand side of eachdepicted structure is bound to the -(CHz)a-. In embodiments,A1is wherein the left hand side of each depicted structure is bound to the -(CHz)a-. In embodiments,A1isO wherein the left hand side of each depicted structure is bound to the -(CHz)a-. In embodiments, A1is wherein the left hand side of each depictedstructure is bound to the -(CHz)a-. In embodiments, A1is wherein the left hand side of each depicted structure is bound to the -(CH2)a-. In embodiments, A1is -O-. In embodiments, A1is -S-. In embodiments, A1is -(C1-C6alkylene)-. In embodiments, A1is -(C3-C5 alkylene)-.

[0127] In embodiments,Z1is, wherein the right hand side of each depictedstructure is bound to the -(CH2)a-. In embodiments, Z1is, wherein the right hand side of each depicted structure is bound to the -(CH2)a-. In embodiments, Z1iswherein the right hand side of each depicted structure is bound to the -(CH2)a-

[0128] In embodiments, Z1is -S-S-.

[0129] In embodiments, R5is hydrogen. In embodiments, R5is optionally substituted (Ci-Cs) alkyl. In embodiments, R5is methyl.

[0130] In embodiments, R6is optionally substituted (Ci-Cs) alkyl. In embodiments, R6is methyl.

[0131] In embodiments, R5and R6are the same. In embodiments, R5and R6are different. In embodiments, R5and R6are both methyl. In embodiments, R5is hydrogen and R6is methyl.

[0132] In embodiments, each R is independently selected from, wherein each R1is independently selected from optionally substituted alkyl, optionally substituted alkenyl, and optionally substituted alkynyl.

[0133] In embodiments, each R1is the same. In embodiments, at least one R1is different.

[0134] In embodiments, each R1is independently selected from optionally substituted (C5-C50) alkyl, optionally substituted (C5-C50) alkenyl, and optionally substituted (C5-C50) alkynyl. In embodiments, each R1is independently selected from optionally substituted (C5-C40) alkyl, optionally substituted (C5-C40) alkenyl, and optionally substituted (C5-C40) alkynyl. In embodiments, each R1is independently selected from optionally substituted (C3-C30) alkyl, optionally substituted (C3-C30) alkenyl, and optionally substituted (C3-C30) alkynyl. In embodiments, each R1is independently selected from optionally substituted (C5-C30) alkyl, optionally substituted (C5-C30) alkenyl, and optionally substituted (C5-C30) alkynyl. In embodiments, each R1is independently selected from optionally substituted (C5-C25) alkyl, optionally substituted (C5-C25) alkenyl, and optionally substituted (C5-C25) alkynyl. In embodiments, each R1is independently selected from optionally substituted (C10-C20) alkyl, optionally substituted (C10-C20) alkenyl, and optionally substituted (C10-C20) alkynyl. In embodiments, each R1is independently selected from optionally substituted (C10-C18) alkyl, optionally substituted (C5-C15) alkenyl, for example optionally substituted (Cs-Cio) alkenyl, and optionally substituted (C5-C15) alkynyl, for example optionally substituted (C8-C10) alkynyl.

[0135] In embodiments, each R1is independently selected from optionally substituted alkyl. In embodiments, each R1is independently selected from optionally substituted (C5-C50) alkyl. In embodiments, each R1is independently selected from optionally substituted (C5-C40) alkyl. In embodiments, each R1is independently selected from optionally substituted (C3-C30) alkyl. In embodiments, each R1is independently selected from optionally substituted (C5-C30) alkyl. In embodiments, each R1is independently selected from optionally substituted (C5-C25) alkyl. In embodiments, each R1is independently selected from optionally substituted (C10-C20) alkyl. In embodiments, each R1is independently selected from optionally substituted (C10-C18) alkyl.

[0136] In embodiments, each R1is independently selected from optionally substituted alkenyl. In embodiments, each R1is independently selected from optionally substituted (C5-C50) alkenyl. In embodiments, each R1is independently selected from optionally substituted (C5-C40) alkenyl. In embodiments, each R1is independently selected from optionally substituted (C3-C30) alkenyl. In embodiments, each R1is independently selected from optionally substituted (C5-C30) alkenyl. In embodiments, each R1is independently selected from optionally substituted (C5-C25) alkenyl. In embodiments, each R1is independently selected from optionally substituted (C10-C20) alkenyl. In embodiments, each R1is independently selected from optionally substituted (C5-C15) alkenyl. In embodiments, each R1is independently selected from optionally substituted (C8-C10) alkenyl.

[0137] In embodiments, each R1is independently selected from optionally substituted alkynyl. In embodiments, each R1is independently selected from optionally substituted (C5-C50) alkynyl. In embodiments, each R1is independently selected from optionally substituted (C5-C40) alkynyl. In embodiments, each R1is independently selected from optionally substituted (C3-C30) alkynyl. In embodiments, each R1is independently selected from optionally substituted (C5-C30) alkynyl. In embodiments, each R1is independently selected from optionally substituted (C5-C25) alkynyl. In embodiments, each R1is independently selected from optionally substituted (C10-C20) alkynyl. In embodiments, each R1is independently selected from optionally substituted (C5-C15) alkynyl. In embodiments, each R1is independently selected from optionally substituted (C8-C10) alkynyl.

[0138] In embodiments, each R1is independently selected from optionally substituted (C5-C25) alkyl, optionally substituted (C5-C25) alkenyl, and optionally substituted (C5-C25) alkynyl.

[0139] In embodiments, each R1is independently selected from optionally substituted alkyl and optionally substituted alkenyl.

[0140] In embodiments, each R1is independently selected from optionally substituted (C5-C30) alkyl and optionally substituted (C5-C30) alkenyl.

[0141] In embodiments, each R1is independently selected from optionally substituted (C5-C25) alkyl and optionally substituted (C5-C25) alkenyl, for example optionally substituted (C10-C18) alkyl and optionally substituted (C8-C10) alkenyl.

[0142] In embodiments, each R1is independently selected from:, optionally wherein each R1is independently selected from options (i), (ii), and (iii).

[0143] In embodiments, each R1is independently selected from:>2

[0144] In embodiments, each R is independently selected from, wherein each R2is independently selected from optionally substituted alkyl, optionally substituted alkenyl, optionally substituted alkynyl, and -W1-X1,wherein each W1is independently selected from optionally substituted alkylene and optionally substituted alkenylene, andeach X1is independently selected from -*O-(C=O)-optionally substituted alkyl, -(*C=O)-O- optionally substituted alkyl, -*O-(C=O)-optionally substituted alkenyl, and -(*C=O)-O-optionally substituted alkenyl, wherein the atom marked with a * is connected to W1.

[0145] In embodiments, each R2A, R2B, R2Cand R2Dis independently selected from optionally substituted alkyl, optionally substituted alkenyl, optionally substituted alkynyl, and -W1-X1, wherein each W1is independently selected from optionally substituted alkylene and optionally substituted alkenylene, andeach X1is independently selected from -*O-(C=O)-optionally substituted alkyl, -(*C=O)-O- optionally substituted alkyl, -*O-(C=O)-optionally substituted alkenyl, and -(*C=O)-O-optionally substituted alkenyl, wherein the atom marked with a * is connected to W1.

[0146] In embodiments, each R2is the same. In embodiments, at least one R2is different.

[0147] In embodiments, R2A, R2B, R2Cand R2Dare the same. In embodiments, R2Aand R2Bare the same. In embodiments, R2Cand R2Dare the same. In embodiments, R2Aand R2Dare the same. In embodiments, R2Band R2Care the same.

[0148] In embodiments, R2Aand R2Bare the same and R2Cand R2Dare the same, but wherein R2Aand R2Bare different to R2Cand R2D. In embodiments, R2Aand R2Dare the same and R2Band R2Care the same, but wherein R2Aand R2Dare different to R2Band R2C.

[0149] In embodiments, each R2or each R2A, R2B, R2Cand R2D, when present, is independently selected from optionally substituted alkyl. In embodiments, each R2or each R2A, R2B, R2Cand R2D, when present, is independently selected from optionally substituted (C5-C50) alkyl. In embodiments, each R2or each R2A, R2B, R2Cand R2D, when present, is independently selected from optionally substituted (C5-C40) alkyl. In embodiments, each R2or each R2A, R2B, R2Cand R2D, when present, is independently selected from optionally substituted (C3-C30) alkyl. In embodiments, each R2or each R2A, R2B, R2Cand R2D, when present, is independently selected from optionally substituted (C5-C30) alkyl. In embodiments, each R2or each R2A, R2B, R2Cand R2D, when present, is independently selected from optionally substituted (C5-C25) alkyl. In embodiments, each R2or each R2A, R2B, R2Cand R2D, when present, is independently selected from optionally substituted (C5-C20) alkyl. In embodiments, each R2or each R2A, R2B, R2Cand R2D, when present, is independently selected from optionally substituted (C8-C12) alkyl.

[0150] In embodiments, each R2A, when present, is optionally substituted alkyl. In embodiments, each R2A, when present, is independently selected from optionally substituted (C5-C50) alkyl. In embodiments, each R2A, when present, is independently selected from optionally substituted (C5-C40) alkyl. In embodiments, each R2A, when present, is independently selected from optionally substituted (C3-C30) alkyl. In embodiments, each R2A, when present, is independently selected fromoptionally substituted (C5-C30) alkyl. In embodiments, each R2A, when present, is optionally substituted (C5-C25) alkyl. In embodiments, each R2A, when present, is optionally substituted (C5-C20) alkyl. In embodiments, each R2A, when present, is optionally substituted (C8-C12) alkyl.

[0151] In embodiments, each R2B, when present, is optionally substituted alkyl. In embodiments, each R2B, when present, is independently selected from optionally substituted (C5-C50) alkyl. In embodiments, each R2B, when present, is independently selected from optionally substituted (C5-C40) alkyl. In embodiments, each R2B, when present, is independently selected from optionally substituted (C3-C30) alkyl. In embodiments, each R2B, when present, is independently selected from optionally substituted (C5-C30) alkyl. In embodiments, each R2B, when present, is optionally substituted (C5-C25) alkyl. In embodiments, each R2B, when present, is optionally substituted (C5-C20) alkyl. In embodiments, each R2B, when present, is optionally substituted (C8-C12) alkyl.

[0152] In embodiments, each R2C, when present, is optionally substituted alkyl. In embodiments, each R2C, when present, is independently selected from optionally substituted (C5-C50) alkyl. In embodiments, each R2C, when present, is independently selected from optionally substituted (C5-C40) alkyl. In embodiments, each R2C, when present, is independently selected from optionally substituted (C3-C30) alkyl. In embodiments, each R2C, when present, is independently selected from optionally substituted (C5-C30) alkyl. In embodiments, each R2C, when present, is optionally substituted (C5-C25) alkyl. In embodiments, each R2C, when present, is optionally substituted (C5-C20) alkyl. In embodiments, each R2C, when present, is optionally substituted (C8-C12) alkyl.

[0153] In embodiments, each R2D, when present, is optionally substituted alkyl. In embodiments, each R2D, when present, is independently selected from optionally substituted (C5-C50) alkyl. In embodiments, each R2D, when present, is independently selected from optionally substituted (C5-C40) alkyl. In embodiments, each R2D, when present, is independently selected from optionally substituted (C3-C30) alkyl. In embodiments, each R2D, when present, is independently selected from optionally substituted (C5-C30) alkyl. In embodiments, each R2D, when present, is optionally substituted (C5-C25) alkyl. In embodiments, each R2D, when present, is optionally substituted (C5-C20) alkyl. In embodiments, each R2D, when present, is optionally substituted (C8-C12) alkyl.

[0154] In embodiments, each R2or each R2A, R2B, R2Cand R2D, when present, is independently selected from optionally substituted alkenyl. In embodiments, each R2or each R2A, R2B, R2Cand R2D, when present, is independently selected from optionally substituted (C5-C50) alkenyl. In embodiments, each R2or each R2A, R2B, R2Cand R2D, when present, is independently selected from optionally substituted (C5-C40) alkenyl. In embodiments, each R2or each R2A, R2B, R2Cand R2D, when present, is independently selected from optionally substituted (C3-C30) alkenyl. In embodiments, each R2or each R2A, R2B, R2Cand R2D, when present, is independently selected from optionallysubstituted (C5-C30) alkenyl. In embodiments, each R2or each R2A, R2B, R2Cand R2D, when present, is independently selected from optionally substituted (C5-C25) alkenyl.

[0155] In embodiments, each R2A, when present, is optionally substituted alkenyl. In embodiments, each R2A, when present, is independently selected from optionally substituted (C5-C50) alkenyl. In embodiments, each R2A, when present, is independently selected from optionally substituted (C5-C40) alkenyl. In embodiments, each R2A, when present, is independently selected from optionally substituted (C3-C30) alkenyl. In embodiments, each R2A, when present, is independently selected from optionally substituted (C5-C30) alkenyl. In embodiments, each R2A, when present, is optionally substituted (C5-C25) alkenyl.

[0156] In embodiments, each R2B, when present, is optionally substituted alkenyl. In embodiments, each R2B, when present, is independently selected from optionally substituted (C5-C50) alkenyl. In embodiments, each R2B, when present, is independently selected from optionally substituted (C5-C40) alkenyl. In embodiments, each R2B, when present, is independently selected from optionally substituted (C3-C30) alkenyl. In embodiments, each R2B, when present, is independently selected from optionally substituted (C5-C30) alkenyl. In embodiments, each R2B, when present, is optionally substituted (C5-C25) alkenyl.

[0157] In embodiments, each R2C, when present, is optionally substituted alkenyl. In embodiments, each R2C, when present, is independently selected from optionally substituted (C5-C50) alkenyl. In embodiments, each R2C, when present, is independently selected from optionally substituted (C5-C40) alkenyl. In embodiments, each R2C, when present, is independently selected from optionally substituted (C3-C30) alkenyl. In embodiments, each R2C, when present, is independently selected from optionally substituted (C5-C30) alkenyl. In embodiments, each R2C, when present, is optionally substituted (C5-C25) alkenyl.

[0158] In embodiments, each R2D, when present, is optionally substituted alkenyl. In embodiments, each R2D, when present, is independently selected from optionally substituted (C5-C50) alkenyl. In embodiments, each R2D, when present, is independently selected from optionally substituted (C5-C40) alkenyl. In embodiments, each R2D, when present, is independently selected from optionally substituted (C3-C30) alkenyl. In embodiments, each R2D, when present, is independently selected from optionally substituted (C5-C30) alkenyl. In embodiments, each R2D, when present, is optionally substituted (C5-C25) alkenyl.

[0159] In embodiments, each R2or each R2A, R2B, R2Cand R2D, when present, is independently selected from optionally substituted alkynyl. In embodiments, each R2or each R2A, R2B, R2Cand R2D, when present, is independently selected from optionally substituted (C5-C50) alkynyl. In embodiments, each R2or each R2A, R2B, R2Cand R2D, when present, is independently selected fromoptionally substituted (C5-C40) alkynyl. In embodiments, each R2or each R2A, R2B, R2Cand R2D, when present, is independently selected from optionally substituted (C3-C30) alkynyl. In embodiments, each R2or each R2A, R2B, R2Cand R2D, when present, is independently selected from optionally substituted (C5-C30) alkynyl. In embodiments, each R2or each R2A, R2B, R2Cand R2D, when present, is independently selected from optionally substituted (C5-C25) alkynyl.

[0160] In embodiments, each R2A, when present, is optionally substituted alkynyl. In embodiments, each R2A, when present, is independently selected from optionally substituted (C5-C50) alkynyl. In embodiments, each R2A, when present, is independently selected from optionally substituted (C5-C40) alkynyl. In embodiments, each R2A, when present, is independently selected from optionally substituted (C3-C30) alkynyl. In embodiments, each R2A, when present, is independently selected from optionally substituted (C5-C30) alkynyl. In embodiments, each R2A, when present, is optionally substituted (C5-C25) alkynyl.

[0161] In embodiments, each R2B, when present, is optionally substituted alkynyl. In embodiments, each R2B, when present, is independently selected from optionally substituted (C5-C50) alkynyl. In embodiments, each R2B, when present, is independently selected from optionally substituted (C5-C40) alkynyl. In embodiments, each R2B, when present, is independently selected from optionally substituted (C3-C30) alkynyl. In embodiments, each R2B, when present, is independently selected from optionally substituted (C5-C30) alkynyl. In embodiments, each R2B, when present, is optionally substituted (C5-C25) alkynyl.

[0162] In embodiments, each R2C, when present, is optionally substituted alkynyl. In embodiments, each R2C, when present, is independently selected from optionally substituted (C5-C50) alkynyl. In embodiments, each R2C, when present, is independently selected from optionally substituted (C5-C40) alkynyl. In embodiments, each R2C, when present, is independently selected from optionally substituted (C3-C30) alkynyl. In embodiments, each R2C, when present, is independently selected from optionally substituted (C5-C30) alkynyl. In embodiments, each R2C, when present, is optionally substituted (C5-C25) alkynyl.

[0163] In embodiments, each R2D, when present, is optionally substituted alkynyl. In embodiments, each R2D, when present, is independently selected from optionally substituted (C5-C50) alkynyl. In embodiments, each R2D, when present, is independently selected from optionally substituted (C5-C40) alkynyl. In embodiments, each R2D, when present, is independently selected from optionally substituted (C3-C30) alkynyl. In embodiments, each R2D, when present, is independently selected from optionally substituted (C5-C30) alkynyl. In embodiments, each R2D, when present, is optionally substituted (C5-C25) alkynyl.

[0164] In embodiments, each R2or each R2A, R2B, R2Cand R2D, when present, is independently selected from -W1-X1.

[0165] In embodiments, each R2A, when present, is -W1-X1.

[0166] In embodiments, each R2A, when present, is -W1-X1, wherein each W1is optionally substituted alkylene. In embodiments, each R2A, when present, is -W1-X1, wherein each W1is optionally substituted (C1-C10) alkylene. In embodiments, each R2A, when present, is -W1-X1, wherein each W1is optionally substituted (C1-C6) alkylene. In embodiments, each R2A, when present, is -W1-X1, wherein each W1is optionally substituted (C4-C5) alkylene.

[0167] In embodiments, each R2A, when present, is -W1-X1, wherein each W1is optionally substituted alkenylene. In embodiments, each R2A, when present, is -W1-X1, wherein each W1is optionally substituted (C2-C10) alkenylene. In embodiments, each R2A, when present, is -W1-X1, wherein each W1is optionally substituted (C2-C6) alkenylene. In embodiments, each R2A, when present, is -W1-X1, wherein each W1is optionally substituted (C4-C5) alkenylene.

[0168] In embodiments, each R2A, when present, is -W1-X1, wherein each X1is -*O-(C=O)-optionally substituted alkyl, wherein the atom marked with a * is connected to W1. In embodiments, each R2A, when present, is -W1-X1, wherein each X1is -*O-(C=O)-optionally substituted (C5-C50) alkyl, wherein the atom marked with a * is connected to W1. In embodiments, each R2A, when present, is -W1-X1, wherein each X1is -*O-(C=O)-optionally substituted (C5-C40) alkyl, wherein the atom marked with a * is connected to W1. In embodiments, each R2A, when present, is -W1-X1, wherein each X1is -*O-(C=O)-optionally substituted (C3-C30) alkyl, wherein the atom marked with a * is connected to W1. In embodiments, each R2A, when present, is -W1-X1, wherein each X1is -*O-(C=O)-optionally substituted (C5-C30) alkyl, wherein the atom marked with a * is connected to W1. In embodiments, each R2A, when present, is -W1-X1, wherein each X1is -*O-(C=O)-optionally substituted (C5-C25) alkyl, wherein the atom marked with a * is connected to W1. In embodiments, each R2A, when present, is -W1-X1, wherein each X1is -*O-(C=O)-optionally substituted (C8-C20) alkyl, wherein the atom marked with a * is connected to W1. In embodiments, each R2A, when present, is -W1-X1, wherein each X1is -*O-(C=O)-optionally substituted (C9-C15) alkyl, wherein the atom marked with a * is connected to W1. In embodiments, each R2A, when present, is -W1-X1, wherein each X1is -*O-(C=O)-optionally substituted (C10-C15) alkyl, wherein the atom marked with a * is connected to W1.

[0169] In embodiments, each R2A, when present, is -W1-X1, wherein each X1is -(*C=O)-O-optionally substituted alkyl, wherein the atom marked with a * is connected to W1. In embodiments, each R2A, when present, is -W1-X1, wherein each X1is -(*C=O)-O-optionally substituted (C5-C50) alkyl, wherein the atom marked with a * is connected to W1. In embodiments, each R2A, when present, is -W1-X1, wherein each X1is -(*C=O)-O-optionally substituted (C5-C40) alkyl, wherein the atom marked with a *is connected to W1. In embodiments, each R2A, when present, is -W1-X1, wherein each X1is -(*C=O)-O-optionally substituted (C3-C30) alkyl, wherein the atom marked with a * is connected to W1. In embodiments, each R2A, when present, is -W1-X1, wherein each X1is -(*C=O)-O-optionally substituted (C5-C30) alkyl, wherein the atom marked with a * is connected to W1. In embodiments, each R2A, when present, is -W1-X1, wherein each X1is -(*C=O)-O-optionally substituted (C5-C25) alkyl, wherein the atom marked with a * is connected to W1. In embodiments, each R2A, when present, is -W1-X1, wherein each X1is -(*C=O)-O-optionally substituted (C8-C20) alkyl, wherein the atom marked with a * is connected to W1. In embodiments, each R2A, when present, is -W1-X1, wherein each X1is -(*C=O)-O-optionally substituted (C9-C15) alkyl, wherein the atom marked with a * is connected to W1. In embodiments, each R2A, when present, is -W1-X1, wherein each X1is -(*C=O)-O-optionally substituted (C10-C15) alkyl, wherein the atom marked with a * is connected to W1.

[0170] In embodiments, each R2A, when present, is -W1-X1, wherein each X1is -*O-(C=O)-optionally substituted alkenyl, wherein the atom marked with a * is connected to W1. In embodiments, each R2A, when present, is -W1-X1, wherein each X1is -*O-(C=O)-optionally substituted (C5-C50) alkenyl, wherein the atom marked with a * is connected to W1. In embodiments, each R2A, when present, is -W1-X1, wherein each X1is -*O-(C=O)-optionally substituted (C5-C40) alkenyl, wherein the atom marked with a * is connected to W1. In embodiments, each R2A, when present, is -W1-X1, wherein each X1is -*O-(C=O)-optionally substituted (C3-C30) alkenyl, wherein the atom marked with a * is connected to W1. In embodiments, each R2A, when present, is -W1-X1, wherein each X1is -*O-(C=O)-optionally substituted (C5-C30) alkenyl, wherein the atom marked with a * is connected to W1. In embodiments, each R2A, when present, is -W1-X1, wherein each X1is -*O-(C=O)-optionally substituted (C5-C25) alkenyl, wherein the atom marked with a * is connected to W1. In embodiments, each R2A, when present, is -W1-X1, wherein each X1is -*O-(C=O)-optionally substituted (C8-C20) alkenyl, wherein the atom marked with a * is connected to W1. In embodiments, each R2A, when present, is -W1-X1, wherein each X1is -*O-(C=O)-optionally substituted (C9-C10) alkenyl, wherein the atom marked with a * is connected to W1.

[0171] In embodiments, each R2A, when present, is -W1-X1, wherein each X1is -(*C=O)-O-optionally substituted alkenyl, wherein the atom marked with a * is connected to W1. In embodiments, each R2A, when present, is -W1-X1, wherein each X1is -(*C=O)-O-optionally substituted (C5-C50) alkenyl, wherein the atom marked with a * is connected to W1. In embodiments, each R2A, when present, is -W1-X1, wherein each X1is -(*C=O)-O-optionally substituted (C5-C40) alkenyl, wherein the atom marked with a * is connected to W1. In embodiments, each R2A, when present, is -W1-X1, wherein each X1is -(*C=O)-O-optionally substituted (C3-C30) alkenyl, wherein the atom marked with a * is connected to W1. In embodiments, each R2A, when present, is -W1-X1, wherein each X1is -(*C=O)-O-optionally substituted (C5-C30) alkenyl, wherein the atom marked with a * is connected to W1. In embodiments, each R2A, when present, is -W1-X1, wherein each X1is -(*C=O)-O-optionally substituted (C5-C25) alkenyl, wherein the atom marked with a * is connected to W1. In embodiments, each R2A, when present, is -W1-X1, wherein each X1is -(*C=O)-O-optionally substituted (C8-C20) alkenyl, wherein the atom marked with a * is connected to W1. In embodiments, each R2A, when present, is -W1-X1, wherein each X1is -(*C=O)-O-optionally substituted (C9-C10) alkenyl, wherein the atom marked with a * is connected to W1.

[0172] In embodiments, each R2B, when present, is -W1-X1.

[0173] In embodiments, each R2B, when present, is -W1-X1, wherein each W1is optionally substituted alkylene. In embodiments, each R2B, when present, is -W1-X1, wherein each W1is optionally substituted (C1-C10) alkylene. In embodiments, each R2B, when present, is -W1-X1, wherein each W1is optionally substituted (C2-C6) alkylene. In embodiments, each R2B, when present, is -W1-X1, wherein each W1is optionally substituted (C4-C5) alkylene.

[0174] In embodiments, each R2B, when present, is -W1-X1, wherein each W1is optionally substituted alkenylene. In embodiments, each R2B, when present, is -W1-X1, wherein each W1is optionally substituted (C2-C10) alkenylene. In embodiments, each R2B, when present, is -W1-X1, wherein each W1is optionally substituted (C2-C6) alkenylene. In embodiments, each R2B, when present, is -W1-X1, wherein each W1is optionally substituted (C4-C5) alkenylene.

[0175] In embodiments, each R2B, when present, is -W1-X1, wherein each X1is -*O-(C=O)-optionally substituted alkyl, wherein the atom marked with a * is connected to W1. In embodiments, each R2B, when present, is -W1-X1, wherein each X1is -*O-(C=O)-optionally substituted (C5-C50) alkyl, wherein the atom marked with a * is connected to W1. In embodiments, each R2B, when present, is -W1-X1, wherein each X1is -*O-(C=O)-optionally substituted (C5-C40) alkyl, wherein the atom marked with a * is connected to W1. In embodiments, each R2B, when present, is -W1-X1, wherein each X1is -*O-(C=O)-optionally substituted (C3-C30) alkyl, wherein the atom marked with a * is connected to W1. In embodiments, each R2B, when present, is -W1-X1, wherein each X1is -*O-(C=O)-optionally substituted (C5-C30) alkyl, wherein the atom marked with a * is connected to W1. In embodiments, each R2B, when present, is -W1-X1, wherein each X1is -*O-(C=O)-optionally substituted (C5-C25) alkyl, wherein the atom marked with a * is connected to W1. In embodiments, each R2B, when present, is -W1-X1, wherein each X1is -*O-(C=O)-optionally substituted (C8-C20) alkyl, wherein the atom marked with a * is connected to W1. In embodiments, each R2B, when present, is -W1-X1, wherein each X1is -*O-(C=O)-optionally substituted (C9-C15) alkyl, wherein the atom marked with a * is connected to W1. In embodiments, each R2B, when present, is -W1-X1, wherein each X1is -*O-(C=O)-optionally substituted (C10-C15) alkyl, wherein the atom marked with a * is connected to W1.

[0176] In embodiments, each R2B, when present, is -W1-X1, wherein each X1is -(*C=O)-O-optionally substituted alkyl, wherein the atom marked with a * is connected to W1. In embodiments, each R2B, when present, is -W1-X1, wherein each X1is -(*C=O)-O-optionally substituted (C5-C50) alkyl, wherein the atom marked with a * is connected to W1. In embodiments, each R2B, when present, is -W1-X1, wherein each X1is -(*C=O)-O-optionally substituted (C5-C40) alkyl, wherein the atom marked with a * is connected to W1. In embodiments, each R2B, when present, is -W1-X1, wherein each X1is -(*C=O)-O-optionally substituted (C3-C30) alkyl, wherein the atom marked with a * is connected to W1. In embodiments, each R2B, when present, is -W1-X1, wherein each X1is -(*C=O)-O-optionally substituted (C5-C30) alkyl, wherein the atom marked with a * is connected to W1. In embodiments, each R2B, when present, is -W1-X1, wherein each X1is -(*C=O)-O-optionally substituted (C5-C25) alkyl, wherein the atom marked with a * is connected to W1. In embodiments, each R2B, when present, is -W1-X1, wherein each X1is -(*C=O)-O-optionally substituted (C8-C20) alkyl, wherein the atom marked with a * is connected to W1. In embodiments, each R2B, when present, is -W1-X1, wherein each X1is -(*C=O)-O-optionally substituted (C9-C15) alkyl, wherein the atom marked with a * is connected to W1. In embodiments, each R2B, when present, is -W1-X1, wherein each X1is -(*C=O)-O-optionally substituted (C10-C15) alkyl, wherein the atom marked with a * is connected to W1.

[0177] In embodiments, each R2B, when present, is -W1-X1, wherein each X1is -*O-(C=O)-optionally substituted alkenyl, wherein the atom marked with a * is connected to W1. In embodiments, each R2B, when present, is -W1-X1, wherein each X1is -*O-(C=O)-optionally substituted (C5-C50) alkenyl, wherein the atom marked with a * is connected to W1. In embodiments, each R2B, when present, is -W1-X1, wherein each X1is -*O-(C=O)-optionally substituted (C5-C40) alkenyl, wherein the atom marked with a * is connected to W1. In embodiments, each R2B, when present, is -W1-X1, wherein each X1is -*O-(C=O)-optionally substituted (C3-C30) alkenyl, wherein the atom marked with a * is connected to W1. In embodiments, each R2B, when present, is -W1-X1, wherein each X1is -*O-(C=O)-optionally substituted (C5-C30) alkenyl, wherein the atom marked with a * is connected to W1. In embodiments, each R2B, when present, is -W1-X1, wherein each X1is -*O-(C=O)-optionally substituted (C5-C25) alkenyl, wherein the atom marked with a * is connected to W1. In embodiments, each R2B, when present, is -W1-X1, wherein each X1is -*O-(C=O)-optionally substituted (C8-C20) alkenyl, wherein the atom marked with a * is connected to W1. In embodiments, each R2B, when present, is -W1-X1, wherein each X1is -*O-(C=O)-optionally substituted (C9-C10) alkenyl, wherein the atom marked with a * is connected to W1.

[0178] In embodiments, each R2B, when present, is -W1-X1, wherein each X1is -(*C=O)-O-optionally substituted alkenyl, wherein the atom marked with a * is connected to W1. In embodiments, each R2B, when present, is -W1-X1, wherein each X1is -(*C=O)-O-optionally substituted (C5-C50) alkenyl,wherein the atom marked with a * is connected to W1. In embodiments, each R2B, when present, is -W1-X1, wherein each X1is -(*C=O)-O-optionally substituted (C5-C40) alkenyl, wherein the atom marked with a * is connected to W1. In embodiments, each R2B, when present, is -W1-X1, wherein each X1is -(*C=O)-O-optionally substituted (C3-C30) alkenyl, wherein the atom marked with a * is connected to W1. In embodiments, each R2B, when present, is -W1-X1, wherein each X1is -(*C=O)-O-optionally substituted (C5-C30) alkenyl, wherein the atom marked with a * is connected to W1. In embodiments, each R2B, when present, is -W1-X1, wherein each X1is -(*C=O)-O-optionally substituted (C5-C25) alkenyl, wherein the atom marked with a * is connected to W1. In embodiments, each R2B, when present, is -W1-X1, wherein each X1is -(*C=O)-O-optionally substituted (C8-C20) alkenyl, wherein the atom marked with a * is connected to W1. In embodiments, each R2B, when present, is -W1-X1, wherein each X1is -(*C=O)-O-optionally substituted (C9-C10) alkenyl, wherein the atom marked with a * is connected to W1.

[0179] In embodiments, each R2C, when present, is -W1-X1.

[0180] In embodiments, each R2C, when present, is -W1-X1, wherein each W1is optionally substituted alkylene. In embodiments, each R2C, when present, is -W1-X1, wherein each W1is optionally substituted (C1-C10) alkylene. In embodiments, each R2C, when present, is -W1-X1, wherein each W1is optionally substituted (C2-C6) alkylene. In embodiments, each R2C, when present, is -W1-X1, wherein each W1is optionally substituted (C4-C5) alkylene.

[0181] In embodiments, each R2C, when present, is -W1-X1, wherein each W1is optionally substituted alkenylene. In embodiments, each R2C, when present, is -W1-X1, wherein each W1is optionally substituted (C2-C10) alkenylene. In embodiments, each R2C, when present, is -W1-X1, wherein each W1is optionally substituted (C2-C6) alkenylene. In embodiments, each R2C, when present, is -W1-X1, wherein each W1is optionally substituted (C4-C5) alkenylene.

[0182] In embodiments, each R2C, when present, is -W1-X1, wherein each X1is -*O-(C=O)-optionally substituted alkyl, wherein the atom marked with a * is connected to W1. In embodiments, each R2C, when present, is -W1-X1, wherein each X1is -*O-(C=O)-optionally substituted (C5-C50) alkyl, wherein the atom marked with a * is connected to W1. In embodiments, each R2C, when present, is -W1-X1, wherein each X1is -*O-(C=O)-optionally substituted (C5-C40) alkyl, wherein the atom marked with a * is connected to W1. In embodiments, each R2C, when present, is -W1-X1, wherein each X1is -*O-(C=O)-optionally substituted (C3-C30) alkyl, wherein the atom marked with a * is connected to W1. In embodiments, each R2C, when present, is -W1-X1, wherein each X1is -*O-(C=O)-optionally substituted (C5-C30) alkyl, wherein the atom marked with a * is connected to W1. In embodiments, each R2C, when present, is -W1-X1, wherein each X1is -*O-(C=O)-optionally substituted (C5-C25) alkyl, wherein the atom marked with a * is connected to W1. In embodiments, each R2C, when present, is -W1-X1,wherein each X1is -*O-(C=O)-optionally substituted (C8-C20) alkyl, wherein the atom marked with a * is connected to W1. In embodiments, each R2C, when present, is -W1-X1, wherein each X1is -*O-(C=O)-optionally substituted (C9-C15) alkyl, wherein the atom marked with a * is connected to W1. In embodiments, each R2C, when present, is -W1-X1, wherein each X1is -*O-(C=O)-optionally substituted (C10-C15) alkyl, wherein the atom marked with a * is connected to W1.

[0183] In embodiments, each R2C, when present, is -W1-X1, wherein each X1is -(*C=O)-O-optionally substituted alkyl, wherein the atom marked with a * is connected to W1. In embodiments, each R2C, when present, is -W1-X1, wherein each X1is -(*C=O)-O-optionally substituted (C5-C50) alkyl, wherein the atom marked with a * is connected to W1. In embodiments, each R2C, when present, is -W1-X1, wherein each X1is -(*C=O)-O-optionally substituted (C5-C40) alkyl, wherein the atom marked with a * is connected to W1. In embodiments, each R2C, when present, is -W1-X1, wherein each X1is -(*C=O)-O-optionally substituted (C3-C30) alkyl, wherein the atom marked with a * is connected to W1. In embodiments, each R2C, when present, is -W1-X1, wherein each X1is -(*C=O)-O-optionally substituted (C5-C30) alkyl, wherein the atom marked with a * is connected to W1. In embodiments, each R2C, when present, is -W1-X1, wherein each X1is -(*C=O)-O-optionally substituted (C5-C25) alkyl, wherein the atom marked with a * is connected to W1. In embodiments, each R2C, when present, is -W1-X1, wherein each X1is -(*C=O)-O-optionally substituted (C8-C20) alkyl, wherein the atom marked with a * is connected to W1. In embodiments, each R2C, when present, is -W1-X1, wherein each X1is -(*C=O)-O-optionally substituted (C9-C15) alkyl, wherein the atom marked with a * is connected to W1. In embodiments, each R2C, when present, is -W1-X1, wherein each X1is -(*C=O)-O-optionally substituted (C10-C15) alkyl, wherein the atom marked with a * is connected to W1.

[0184] In embodiments, each R2C, when present, is -W1-X1, wherein each X1is -*O-(C=O)-optionally substituted alkenyl, wherein the atom marked with a * is connected to W1. In embodiments, each R2C, when present, is -W1-X1, wherein each X1is -*O-(C=O)-optionally substituted (C5-C50) alkenyl, wherein the atom marked with a * is connected to W1. In embodiments, each R2C, when present, is -W1-X1, wherein each X1is -*O-(C=O)-optionally substituted (C5-C40) alkenyl, wherein the atom marked with a * is connected to W1. In embodiments, each R2C, when present, is -W1-X1, wherein each X1is -*O-(C=O)-optionally substituted (C3-C30) alkenyl, wherein the atom marked with a * is connected to W1. In embodiments, each R2C, when present, is -W1-X1, wherein each X1is -*O-(C=O)-optionally substituted (C5-C30) alkenyl, wherein the atom marked with a * is connected to W1. In embodiments, each R2C, when present, is -W1-X1, wherein each X1is -*O-(C=O)-optionally substituted (C5-C25) alkenyl, wherein the atom marked with a * is connected to W1. In embodiments, each R2C, when present, is -W1-X1, wherein each X1is -*O-(C=O)-optionally substituted (C8-C20) alkenyl, wherein the atom marked with a * is connected to W1. In embodiments, each R2C, when present,is -W1-X1, wherein each X1is -*O-(C=O)-optionally substituted (Cs-Cio) alkenyl, wherein the atom marked with a * is connected to W1.

[0185] In embodiments, each R2C, when present, is -W1-X1, wherein each X1is -(*C=O)-O-optionally substituted alkenyl, wherein the atom marked with a * is connected to W1. In embodiments, each R2C, when present, is -W1-X1, wherein each X1is -(*C=O)-O-optionally substituted (C5-C50) alkenyl, wherein the atom marked with a * is connected to W1. In embodiments, each R2C, when present, is -W1-X1, wherein each X1is -(*C=O)-O-optionally substituted (C5-C40) alkenyl, wherein the atom marked with a * is connected to W1. In embodiments, each R2C, when present, is -W1-X1, wherein each X1is -(*C=O)-O-optionally substituted (C3-C30) alkenyl, wherein the atom marked with a * is connected to W1. In embodiments, each R2C, when present, is -W1-X1, wherein each X1is -(*C=O)-O-optionally substituted (C5-C30) alkenyl, wherein the atom marked with a * is connected to W1. In embodiments, each R2C, when present, is -W1-X1, wherein each X1is -(*C=O)-O-optionally substituted (C5-C25) alkenyl, wherein the atom marked with a * is connected to W1. In embodiments, each R2C, when present, is -W1-X1, wherein each X1is -(*C=O)-O-optionally substituted (C8-C20) alkenyl, wherein the atom marked with a * is connected to W1. In embodiments, each R2C, when present, is -W1-X1, wherein each X1is -(*C=O)-O-optionally substituted (Cs-Cio) alkenyl, wherein the atom marked with a * is connected to W1.

[0186] In embodiments, each R2D, when present, is -W1-X1.

[0187] In embodiments, each R2D, when present, is -W1-X1, wherein each W1is optionally substituted alkylene. In embodiments, each R2D, when present, is -W1-X1, wherein each W1is optionally substituted (C1-C10) alkylene. In embodiments, each R2D, when present, is -W1-X1, wherein each W1is optionally substituted (C2-C6) alkylene. In embodiments, each R2D, when present, is -W1-X1, wherein each W1is optionally substituted (C4-C5) alkylene.

[0188] In embodiments, each R2D, when present, is -W1-X1, wherein each W1is optionally substituted alkenylene. In embodiments, each R2D, when present, is -W1-X1, wherein each W1is optionally substituted (C2-C10) alkenylene. In embodiments, each R2D, when present, is -W1-X1, wherein each W1is optionally substituted (C2-C6) alkenylene. In embodiments, each R2D, when present, is -W1-X1, wherein each W1is optionally substituted (C4-C5) alkenylene.

[0189] In embodiments, each R2D, when present, is -W1-X1, wherein each X1is -*O-(C=O)-optionally substituted alkyl, wherein the atom marked with a * is connected to W1. In embodiments, each R2D, when present, is -W1-X1, wherein each X1is -*O-(C=O)-optionally substituted (C5-C50) alkyl, wherein the atom marked with a * is connected to W1. In embodiments, each R2D, when present, is -W1-X1, wherein each X1is -*O-(C=O)-optionally substituted (C5-C40) alkyl, wherein the atom marked with a * is connected to W1. In embodiments, each R2D, when present, is -W1-X1, wherein each X1is -*O-(C=O)-optionally substituted (C3-C30) alkyl, wherein the atom marked with a * is connected to W1. In embodiments, each R2D, when present, is -W1-X1, wherein each X1is -*O-(C=O)-optionally substituted (C5-C30) alkyl, wherein the atom marked with a * is connected to W1. In embodiments, each R2D, when present, is -W1-X1, wherein each X1is -*O-(C=O)-optionally substituted (C5-C25) alkyl, wherein the atom marked with a * is connected to W1. In embodiments, each R2D, when present, is -W1-X1, wherein each X1is -*O-(C=O)-optionally substituted (C8-C20) alkyl, wherein the atom marked with a * is connected to W1. In embodiments, each R2D, when present, is -W1-X1, wherein each X1is -*O-(C=O)-optionally substituted (Cs-Cis) alkyl, wherein the atom marked with a * is connected to W1. In embodiments, each R2D, when present, is -W1-X1, wherein each X1is -*O-(C=O)-optionally substituted (Cio-Cis) alkyl, wherein the atom marked with a * is connected to W1.

[0190] In embodiments, each R2D, when present, is -W1-X1, wherein each X1is -(*C=O)-O-optionally substituted alkyl, wherein the atom marked with a * is connected to W1. In embodiments, each R2D, when present, is -W1-X1, wherein each X1is -(*C=O)-O-optionally substituted (C5-C50) alkyl, wherein the atom marked with a * is connected to W1. In embodiments, each R2D, when present, is -W1-X1, wherein each X1is -(*C=O)-O-optionally substituted (C5-C40) alkyl, wherein the atom marked with a * is connected to W1. In embodiments, each R2D, when present, is -W1-X1, wherein each X1is -(*C=O)-O-optionally substituted (C3-C30) alkyl, wherein the atom marked with a * is connected to W1. In embodiments, each R2D, when present, is -W1-X1, wherein each X1is -(*C=O)-O-optionally substituted (C5-C30) alkyl, wherein the atom marked with a * is connected to W1. In embodiments, each R2D, when present, is -W1-X1, wherein each X1is -(*C=O)-O-optionally substituted (C5-C25) alkyl, wherein the atom marked with a * is connected to W1. In embodiments, each R2D, when present, is -W1-X1, wherein each X1is -(*C=O)-O-optionally substituted (C8-C20) alkyl, wherein the atom marked with a * is connected to W1. In embodiments, each R2D, when present, is -W1-X1, wherein each X1is -(*C=O)-O-optionally substituted (Cs-Cis) alkyl, wherein the atom marked with a * is connected to W1. In embodiments, each R2D, when present, is -W1-X1, wherein each X1is -(*C=O)-O-optionally substituted (Cio-Cis) alkyl, wherein the atom marked with a * is connected to W1.

[0191] In embodiments, each R2D, when present, is -W1-X1, wherein each X1is -*O-(C=O)-optionally substituted alkenyl, wherein the atom marked with a * is connected to W1. In embodiments, each R2D, when present, is -W1-X1, wherein each X1is -*O-(C=O)-optionally substituted (C5-C50) alkenyl, wherein the atom marked with a * is connected to W1. In embodiments, each R2D, when present, is -W1-X1, wherein each X1is -*O-(C=O)-optionally substituted (C5-C40) alkenyl, wherein the atom marked with a * is connected to W1. In embodiments, each R2D, when present, is -W1-X1, wherein each X1is -*O-(C=O)-optionally substituted (C3-C30) alkenyl, wherein the atom marked with a * is connected to W1. In embodiments, each R2D, when present, is -W1-X1, wherein each X1is -*O-(C=O)-optionally substituted (C5-C30) alkenyl, wherein the atom marked with a * is connected to W1. In embodiments, each R2D, when present, is -W1-X1, wherein each X1is -*O-(C=O)-optionally substituted (C5-C25) alkenyl, wherein the atom marked with a * is connected to W1. In embodiments, each R2D, when present, is -W1-X1, wherein each X1is -*O-(C=O)-optionally substituted (C8-C20) alkenyl, wherein the atom marked with a * is connected to W1. In embodiments, each R2D, when present, is -W1-X1, wherein each X1is -*O-(C=O)-optionally substituted (Cs-Cio) alkenyl, wherein the atom marked with a * is connected to W1.

[0192] In embodiments, each R2D, when present, is -W1-X1, wherein each X1is -(*C=O)-O-optionally substituted alkenyl, wherein the atom marked with a * is connected to W1. In embodiments, each R2D, when present, is -W1-X1, wherein each X1is -(*C=O)-O-optionally substituted (C5-C50) alkenyl, wherein the atom marked with a * is connected to W1. In embodiments, each R2D, when present, is -W1-X1, wherein each X1is -(*C=O)-O-optionally substituted (C5-C40) alkenyl, wherein the atom marked with a * is connected to W1. In embodiments, each R2D, when present, is -W1-X1, wherein each X1is -(*C=O)-O-optionally substituted (C3-C30) alkenyl, wherein the atom marked with a * is connected to W1. In embodiments, each R2D, when present, is -W1-X1, wherein each X1is -(*C=O)-O-optionally substituted (C5-C30) alkenyl, wherein the atom marked with a * is connected to W1. In embodiments, each R2D, when present, is -W1-X1, wherein each X1is -(*C=O)-O-optionally substituted (C5-C25) alkenyl, wherein the atom marked with a * is connected to W1. In embodiments, each R2D, when present, is -W1-X1, wherein each X1is -(*C=O)-O-optionally substituted (C8-C20) alkenyl, wherein the atom marked with a * is connected to W1. In embodiments, each R2D, when present, is -W1-X1, wherein each X1is -(*C=O)-O-optionally substituted (Cs-Cio) alkenyl, wherein the atom marked with a * is connected to W1.

[0193] In embodiments, each R2or each R2A, R2B, R2Cand R2D, when present, is independently selected from optionally substituted (C3-C30) alkyl, optionally substituted (C5-C25) alkenyl, optionally substituted (C5-C25) alkynyl, and -W1-X1,wherein each W1is independently selected from optionally substituted (C1-C10) alkylene and optionally substituted (C2-C10) alkenylene, andeach X1is independently selected from -*O-(C=O)-optionally substituted (C5-C25) alkyl, -(*C=O)-O- optionally substituted (C5-C25) alkyl, -*O-(C=O)-optionally substituted (C5-C25) alkenyl, and - (*C=O)-O-optionally substituted (C5-C25) alkenyl, wherein the atom marked with a * is connected to W1.

[0194] In embodiments, each R2or each R2A, R2B, R2Cand R2D, when present, is independently selected from optionally substituted (C3-C28) alkyl, optionally substituted (C3-C28) alkenyl, optionally substituted (C3-C28) alkynyl, and -W1-X1,wherein each W1is independently selected from optionally substituted (C3-C5) alkylene and optionally substituted (C3-C5) alkenylene, andeach X1is independently selected from -*O-(C=O)-optionally substituted (C5-C30) alkyl, -(*C=O)-O- optionally substituted (C5-C30) alkyl, -*O-(C=O)-optionally substituted (C5-C30) alkenyl, and - (*C=O)-O-optionally substituted (C5-C30) alkenyl, wherein the atom marked with a * is connected to W1.

[0195] In embodiments, each R2or each R2A, R2B, R2Cand R2D, when present, is independently selected from optionally substituted (C3-C28) alkyl, optionally substituted (C3-C28) alkenyl, optionally substituted (C3-C28) alkynyl, and -W1-X1,wherein each W1is independently selected from optionally substituted (C3-C5) alkylene and optionally substituted (C3-C5) alkenylene, andeach X1is independently selected from -(*C=O)-O-optionally substituted (C5-C30) alkyl, and - (*C=O)-O-optionally substituted (C5-C30) alkenyl, wherein the atom marked with a * is connected to W1.

[0196] In embodiments, each R2or each R2A, R2B, R2Cand R2D, when present, is independently selected from optionally substituted (C5-C25) alkyl, optionally substituted (C5-C25) alkenyl, optionally substituted (C5-C25) alkynyl, and -W1-X1,wherein each W1is independently selected from optionally substituted (C5-C6) alkylene and optionally substituted (C5-C6) alkenylene, andeach X1is independently selected from -*O-(C=O)-optionally substituted (C5-C6) alkyl, -(*C=O)-O- optionally substituted (C5-C6) alkyl, -*O-(C=O)-optionally substituted (C5-C6) alkenyl, and -(*C=O)- O-optionally substituted (C5-C6) alkenyl, wherein the atom marked with a * is connected to W1.

[0197] In embodiments, each R2or each R2A, R2B, R2Cand R2D, when present, is independently selected from optionally substituted alkyl and -W1-X1, optionally whereineach W1is independently selected from optionally substituted alkylene, andeach X1is independently selected from -*O-(C=O)-optionally substituted alkyl, and -(*C=O)-O- optionally substituted alkenyl, wherein the atom marked with a * is connected to W1.

[0198] In embodiments, each R2or each R2A, R2B, R2Cand R2D, when present, is independently selected from optionally substituted (C5-C25) alkyl, for example optionally substituted (C8-C12) alkyl, and -W1-X1,wherein each W1is independently selected from optionally substituted (C1-C10) alkylene, for example optionally substituted (C4-C5) alkylene, and optionally substituted (C2-C10) alkenylene, for example optionally substituted (C4-C5) alkenylene, andeach X1is independently selected from -*O-(C=O)-optionally substituted (C5-C25) alkyl, for example -*O-(C=O)-optionally substituted (C10-C18) alkyl, -(*C=O)-O-optionally substituted (C5-C25) alkyl, for example -(*C=O)-O-optionally substituted (Cio-Cis) alkyl, -*O-(C=O)-optionally substituted (C5-C25) alkenyl, for example -*O-(C=O)-optionally substituted (Cs-Cio) alkenyl, and - (*C=O)-O-optionally substituted (C5-C25) alkenyl, for example -(*C=O)-O-optionally substituted (Cs-Cio) alkenyl, wherein the atom marked with a * is connected to W1.

[0199] In embodiments, each R2or each R2A, R2B, R2Cand R2D, when present, is independently selected from optionally substituted alkyl or optionally substituted alkenyl. In embodiments, each R2or each R2A, R2B, R2Cand R2D, when present, is independently selected from optionally substituted (C5-C25) alkyl or optionally substituted (C5-C25) alkenyl. In embodiments, each R2or each R2A, R2B, R2Cand R2D, when present, is independently selected from optionally substituted alkyl. In embodiments, each R2or each R2A, R2B, R2Cand R2D, when present, is independently selected from optionally substituted (C5-C25) alkyl, for example optionally substituted (C8-C12) alkyl.

[0200] In embodiments, each R2or each R2A, R2B, R2Cand R2D, when present, is independently selected from -W1-X1, optionally whereineach W1is independently selected from optionally substituted alkylene, andeach X1is independently selected from -*O-(C=O)-optionally substituted alkyl, and -(*C=O)-O- optionally substituted alkenyl, wherein the atom marked with a * is connected to W1.

[0201] In embodiments, each R2or each R2A, R2B, R2Cand R2D, when present, is independently selected from -W1-X1, optionally whereineach W1is independently selected from optionally substituted (C1-C10) alkylene, for example optionally substituted (C4-C5) alkylene, andeach X1is independently selected from -*O-(C=O)-optionally substituted (C5-C25) alkyl, for example -*O-(C=O)-optionally substituted (C10-C18) alkyl, and -(*C=O)-O-optionally substituted (C5-C25) alkenyl, for example -(*C=O)-O-optionally substituted (Cs-Cio) alkenyl, wherein the atom marked with a * is connected to W1.

[0202] In embodiments, each R2or each R2A, R2B, R2Cand R2D, when present, is -W1-X1, wherein each W1is independently selected from optionally substituted (C4-C6) alkylene, and each X1is independently selected from -(*C=O)-O-optionally substituted (C4-C6) alkyl, wherein the atom marked with a * is connected to W1.

[0203] In embodiments, each R2or each R2A, R2B, R2Cand R2D, when present, is independently selected from:o

[0204] In embodiments, each R2or each R2A, R2B, R2Cand R2D, when present, is independently selected from:(ii)(iii)(xiii), optionally wherein each R2or each R2A, R2B, R2Cand R2D, when present, is independently selected from options (i), (ii), (viii), (x), and (xi); or wherein each R2or each R2A, R2B, R2Cand R2D, when present, is independently selected from options (ii), (viii), (x), and (xi).

[0205] In embodiments, the optionally substituted alkyl in each X1is independently selected from:

[0206] In embodiments, each R2or each R2A, R2B, R2Cand R2D, when present, is independently selected from:O, wherein each R3is independently selected from optionally substituted alkyl, optionally substituted alkenyl, and optionally substituted alkynyl.

[0208] In embodiments, each R3is the same. In embodiments, at least one R3is different.

[0209] In embodiments, each R3is independently selected from optionally substituted (C5-C50) alkyl, optionally substituted (C5-C50) alkenyl, and optionally substituted (C5-C50) alkynyl. In embodiments, each R3is independently selected from optionally substituted (C5-C40) alkyl, optionally substituted (C5-C40) alkenyl, and optionally substituted (C5-C40) alkynyl. In embodiments, each R3is independently selected from optionally substituted (C3-C30) alkyl, optionally substituted (C3-C30) alkenyl, and optionally substituted (C3-C30) alkynyl. In embodiments, each R3is independently selected from optionally substituted (C5-C30) alkyl, optionally substituted (C5-C30) alkenyl, and optionally substituted (C5-C30) alkynyl. In embodiments, each R3is independently selected from optionally substituted (C5-C25) alkyl, optionally substituted (C5-C25) alkenyl, and optionally substituted (C5-C25) alkynyl. In embodiments, each R3is independently selected from optionally substituted (C10-C20) alkyl, optionally substituted (C10-C20) alkenyl, and optionally substituted (C10-C20) alkynyl. In embodiments, each R3is independently selected from optionally substituted (C10-C18) alkyl, optionally substituted (C5-C15) alkenyl, for example optionally substituted (Cs-Cio) alkenyl, and optionally substituted (C5-Ci5) alkynyl, for example optionally substituted (C8-C10) alkynyl.

[0210] In embodiments, each R3is independently selected from optionally substituted alkyl. In embodiments, each R3is independently selected from optionally substituted (C5-C50) alkyl. In embodiments, each R3is independently selected from optionally substituted (C5-C40) alkyl. In embodiments, each R3is independently selected from optionally substituted (C3-C30) alkyl. In embodiments, each R3is independently selected from optionally substituted (C5-C30) alkyl. In embodiments, each R3is independently selected from optionally substituted (C5-C25) alkyl. In embodiments, each R3is independently selected from optionally substituted (C10-C20) alkyl. In embodiments, each R3is independently selected from optionally substituted (C10-C18) alkyl.

[0211] In embodiments, each R3is independently selected from optionally substituted alkenyl. In embodiments, each R3is independently selected from optionally substituted (C5-C50) alkenyl. In embodiments, each R3is independently selected from optionally substituted (C5-C40) alkenyl. In embodiments, each R3is independently selected from optionally substituted (C3-C30) alkenyl. In embodiments, each R3is independently selected from optionally substituted (C5-C30) alkenyl. In embodiments, each R3is independently selected from optionally substituted (C5-C25) alkenyl. In embodiments, each R3is independently selected from optionally substituted (C10-C20) alkenyl. In embodiments, each R3is independently selected from optionally substituted (C5-C15) alkenyl. In embodiments, each R3is independently selected from optionally substituted (C8-C10) alkenyl.

[0212] In embodiments, each R3is independently selected from optionally substituted alkynyl. In embodiments, each R3is independently selected from optionally substituted (C5-C50) alkynyl. In embodiments, each R3is independently selected from optionally substituted (C5-C40) alkynyl. Inembodiments, each R3is independently selected from optionally substituted (C3-C30) alkynyl. In embodiments, each R3is independently selected from optionally substituted (C5-C30) alkynyl. In embodiments, each R3is independently selected from optionally substituted (C5-C25) alkynyl. In embodiments, each R3is independently selected from optionally substituted (C10-C20) alkynyl. In embodiments, each R3is independently selected from optionally substituted (C5-C15) alkynyl. In embodiments, each R3is independently selected from optionally substituted (C8-C10) alkynyl.

[0213] In embodiments, each R3is independently selected from optionally substituted (C5-C25) alkyl, optionally substituted (C5-C25) alkenyl, and optionally substituted (C5-C25) alkynyl.

[0214] In embodiments, each R3is independently selected from optionally substituted alkyl.

[0215] In embodiments, each R3is independently selected from optionally substituted (C5-C25) alkyl, for example optionally substituted (C10-C18) alkyl.

[0216] In embodiments, each R3is independently selected from:, optionally wherein each R3is option (i).

[0217] In embodiments, each R3is independently selected from

[0218] In embodiments, each R is independently selected from:1, wherein each R4is independently selected from optionally substituted alkyl, optionally substituted alkenyl, and optionally substituted alkynyl.

[0219] In embodiments, each R4is the same. In embodiments, at least one R4is different.

[0220] In embodiments, each R4is independently selected from optionally substituted (C5-C50) alkyl, optionally substituted (C5-C50) alkenyl, and optionally substituted (C5-C50) alkynyl. In embodiments,each R4is independently selected from optionally substituted (C5-C40) alkyl, optionally substituted (C5-C40) alkenyl, and optionally substituted (C5-C40) alkynyl. In embodiments, each R4is independently selected from optionally substituted (C3-C30) alkyl, optionally substituted (C3-C30) alkenyl, and optionally substituted (C3-C30) alkynyl. In embodiments, each R4is independently selected from optionally substituted (C5-C30) alkyl, optionally substituted (C5-C30) alkenyl, and optionally substituted (C5-C30) alkynyl. In embodiments, each R4is independently selected from optionally substituted (C5-C25) alkyl, optionally substituted (C5-C25) alkenyl, and optionally substituted (C5-C25) alkynyl. In embodiments, each R4is independently selected from optionally substituted (C10-C20) alkyl, optionally substituted (C10-C20) alkenyl, and optionally substituted (C10-C20) alkynyl. In embodiments, each R4is independently selected from optionally substituted (C10-C18) alkyl, optionally substituted (C5-C15) alkenyl, for example optionally substituted (C8-C10) alkenyl, and optionally substituted (C5-C15) alkynyl, for example optionally substituted (C8-C10) alkynyl.

[0221] In embodiments, each R4is independently selected from optionally substituted alkyl. In embodiments, each R4is independently selected from optionally substituted (C5-C50) alkyl. In embodiments, each R4is independently selected from optionally substituted (C5-C40) alkyl. In embodiments, each R4is independently selected from optionally substituted (C3-C30) alkyl. In embodiments, each R4is independently selected from optionally substituted (C5-C30) alkyl. In embodiments, each R4is independently selected from optionally substituted (C5-C25) alkyl. In embodiments, each R4is independently selected from optionally substituted (C10-C20) alkyl. In embodiments, each R4is independently selected from optionally substituted (C10-C18) alkyl.

[0222] In embodiments, each R4is independently selected from optionally substituted alkenyl. In embodiments, each R4is independently selected from optionally substituted (C5-C50) alkenyl. In embodiments, each R4is independently selected from optionally substituted (C5-C40) alkenyl. In embodiments, each R4is independently selected from optionally substituted (C3-C30) alkenyl. In embodiments, each R4is independently selected from optionally substituted (C5-C30) alkenyl. In embodiments, each R4is independently selected from optionally substituted (C5-C25) alkenyl. In embodiments, each R4is independently selected from optionally substituted (C10-C20) alkenyl. In embodiments, each R4is independently selected from optionally substituted (C5-C15) alkenyl. In embodiments, each R4is independently selected from optionally substituted (C8-C10) alkenyl.

[0223] In embodiments, each R4is independently selected from optionally substituted alkynyl. In embodiments, each R4is independently selected from optionally substituted (C5-C50) alkynyl. In embodiments, each R4is independently selected from optionally substituted (C5-C40) alkynyl. In embodiments, each R4is independently selected from optionally substituted (C3-C30) alkynyl. In embodiments, each R4is independently selected from optionally substituted (C5-C30) alkynyl. Inembodiments, each R4is independently selected from optionally substituted (C5-C25) alkynyl. In embodiments, each R4is independently selected from optionally substituted (C10-C20) alkynyl. In embodiments, each R4is independently selected from optionally substituted (C5-C15) alkynyl. In embodiments, each R4is independently selected from optionally substituted (C8-C10) alkynyl.

[0224] In embodiments, each R4, when present, is independently selected from optionally substituted (C5-C25) alkyl, optionally substituted (C5-C25) alkenyl, and optionally substituted (C5-C25) alkynyl.

[0225] In embodiments, each R is the same. In embodiments, at least one R is different.

[0226] In embodiments, each R is independently selected from optionally substituted alkyl or optionally substituted alkenyl.

[0227] In embodiments, each R is independently selected from optionally substituted (C5-C50) alkyl and optionally substituted (C5-C50) alkenyl. In embodiments, each R is independently selected from optionally substituted (C5-C40) alkyl and optionally substituted (C5-C40) alkenyl. In embodiments, each R is independently selected from optionally substituted (C3-C30) alkyl and optionally substituted (C3-C30) alkenyl. In embodiments, each R is independently selected from optionally substituted (C5-C30) alkyl and optionally substituted (C5-C30) alkenyl. In embodiments, each R is independently selected from optionally substituted (C5-C25) alkyl and optionally substituted (C5-C25) alkenyl. In embodiments, each R is independently selected from optionally substituted (C10-C20) alkyl and optionally substituted (C10-C20) alkenyl. In embodiments, each R is independently selected from optionally substituted (C10-C18) alkyl and optionally substituted (C5-C15) alkenyl, for example optionally substituted (C8-C10) alkenyl.

[0228] In embodiments, each R is independently selected from optionally substituted alkyl. In embodiments, each R is independently selected from optionally substituted (C5-C50) alkyl. In embodiments, each R is independently selected from optionally substituted (C5-C40) alkyl. In embodiments, each R is independently selected from optionally substituted (C3-C30) alkyl. In embodiments, each R is independently selected from optionally substituted (C5-C30) alkyl. In embodiments, each R is independently selected from optionally substituted (C5-C25) alkyl. In embodiments, each R is independently selected from optionally substituted (C10-C20) alkyl. In embodiments, each R is independently selected from optionally substituted (C10-C18) alkyl.

[0229] In embodiments, each R is independently selected from optionally substituted alkenyl. In embodiments, each R is independently selected from optionally substituted (C5-C50) alkenyl. In embodiments, each R is independently selected from optionally substituted (C5-C40) alkenyl. In embodiments, each R is independently selected from optionally substituted (C3-C30) alkenyl. In embodiments, each R is independently selected from optionally substituted (C5-C30) alkenyl. Inembodiments, each R is independently selected from optionally substituted (C5-C25) alkenyl. In embodiments, each R is independently selected from optionally substituted (C10-C20) alkenyl. In embodiments, each R is independently selected from optionally substituted (C5-C15) alkenyl. In embodiments, each R is independently selected from optionally substituted (C8-C10) alkenyl.

[0230] In embodiments, each R is independently selected from optionally substituted (C5-C30) alkyl or optionally substituted (C5-C30) alkenyl.

[0231] In embodiments, each R is independently selected from optionally substituted (C5-C30) alkyl.

[0232] In embodiments, each R is independently selected from optionally substituted C5-C18 alkenyl.

[0233] In embodiments, each R is independently selected from:

[0234] In embodiments, the substituents are not optionally substituted. In embodiments, the optionally substituted substituents are unsubstituted or substituted with one or more halogen (e.g. fluorine). In embodiments, the optionally substituted substituents are substituted with one or more fluorine.

[0235] In embodiments, the compounds of the present invention have any one of the structures in Table 1, or a pharmaceutically acceptable salt thereof.

[0236] In embodiments, provided herein is a composition comprising a compound of the present invention, or a pharmaceutically acceptable salt thereof, and further comprising:(i) one or more non-cationic lipids,(ii) one or more cholesterol-based lipids and(iii) one or more PEG-modified lipids.

[0237] In embodiments, this composition is a lipid nanoparticle, optionally a liposome.

[0238] In embodiments, the one or more cationic lipid(s) constitute(s) about 30 mol %-60 mol % of the lipid nanoparticle. In embodiments, the one or more cationic lipid(s) constitute(s) about 35 mol %-55 mol % of the lipid nanoparticle. In embodiments, the one or more cationic lipid(s) constitute(s) about 40 mol %-50 mol % of the lipid nanoparticle.

[0239] In embodiments, the one or more non-cationic lipid(s) constitute(s) about 10 mol%-50 mol% of the lipid nanoparticle. In embodiments, the one or more non-cationic lipid(s) constitute(s) about 15 mol%-45 mol% of the lipid nanoparticle. In embodiments, the one or more non-cationic lipid(s)constitute(s) about 20 mol%-40 mol% of the lipid nanoparticle. In embodiments, the one or more non-cationic lipid(s) constitute(s) about 25 mol%-35 mol% of the lipid nanoparticle.

[0240] In embodiments, the one or more PEG-modified lipid(s) constitute(s) about 1 mol%-10 mol% of the lipid nanoparticle. In embodiments, the one or more PEG-modified lipid(s) constitute(s) about 2 mol%-9 mol% of the lipid nanoparticle. In embodiments, the one or more PEG-modified lipid(s) constitute(s) about 3 mol%-8 mol% of the lipid nanoparticle. In embodiments, the one or more PEG-modified lipid(s) constitute(s) about 4 mol%-7 mol% of the lipid nanoparticle. In embodiments, the one or more PEG-modified lipid(s) constitute(s) about 5 mol%-6 mol% of the lipid nanoparticle.

[0241] In embodiments, the cholesterol-based lipid constitutes about 10 mol%-50 mol% of the lipid nanoparticle. In embodiments, the cholesterol-based lipid constitutes about 15 mol%-45 mol% of the lipid nanoparticle. In embodiments, the cholesterol-based lipid constitutes about 20 mol%-40 mol% of the lipid nanoparticle. In embodiments, the cholesterol-based lipid constitutes about 25 mol%-35 mol% of the lipid nanoparticle.

[0242] In embodiments, the lipid nanoparticle encapsulates a nucleic acid, optionally an mRNA encoding a peptide or protein. In embodiments, the lipid nanoparticle encapsulates an mRNA encoding a peptide or protein, optionally for use in a vaccine. In embodiments, the peptide encoded by the mRNA is an antigen. In embodiments, the mRNA encodes an influenza antigen. In embodiments, the mRNA encodes an antigen of influenza A virus. In embodiments, the mRNA encodes an antigen of influenza B virus. In embodiments, the mRNA encodes an antigen of Chlamydia sp. bacteria. In embodiments, the mRNA encodes an antigen of C.trachomatis. In embodiments, the mRNA encodes an antigen of P. gingivalis. In embodiments, the mRNA encodes an acne antigen. In embodiments, the mRNA encodes an antigen of C.acnes.

[0243] As used herein, the phrase "encapsulation percentage" refers to the fraction of therapeutic agent (e.g. mRNA) that is effectively encapsulated within a liposomal-based vehicle (e.g. a lipid nanoparticle) relative to the initial fraction of therapeutic agent present in the lipid phase. In embodiments, the lipid nanoparticles have an encapsulation percentage for mRNA of at least 50%. In embodiments, the lipid nanoparticles have an encapsulation percentage for mRNA of at least 55%. In embodiments, the lipid nanoparticles have an encapsulation percentage for mRNA of at least 60%. In embodiments, the lipid nanoparticles have an encapsulation percentage for mRNA of at least 65%. In embodiments, the lipid nanoparticles have an encapsulation percentage for mRNA of at least 70%. In embodiments, the lipid nanoparticles have an encapsulation percentage for mRNA of at least 75%. In embodiments, the lipid nanoparticles have an encapsulation percentage for mRNA of at least 80%. In embodiments, the lipid nanoparticles have an encapsulation percentage for mRNA of at least 85%. In embodiments, the lipid nanoparticles have an encapsulation percentage for mRNAof at least 90%. In embodiments, the lipid nanoparticles have an encapsulation percentage for mRNA of at least 95%. In embodiments, the encapsulation percentage is calculated by performing the Ribogreen assay (Invitrogen) with and without the presence of 0.1% Triton-X 100.

[0244] In embodiments, the composition of the present invention is for use in therapy.

[0245] In embodiments, the composition of the present invention is for use in a method of treating or preventing a disease amenable to treatment or prevention by the peptide or protein encoded by the mRNA encapsulated within said composition.

[0246] In embodiments, the composition of the present invention is for use in a method of treating, preventing or ameliorating a disease, disorder or infection amenable to treatment, prevention or amelioration by the peptide or protein encoded by the mRNA, optionally wherein the disease, disorder or infection is (a) a protein deficiency, optionally wherein the protein deficiency affects the liver, lung, brain or muscle, (b) an autoimmune disease, (c) an infectious disease, or (d) cancer.

[0247] In embodiments, the composition of the present invention is for use in a method of inducing an immune response in a subject.

[0248] In embodiments, the disease or disorder is caused by a viral infection. In embodiments, the disease or disorder is influenza.

[0249] In embodiments, the disease or disorder is caused by a bacterial infection. In embodiments, the disease or disorder is chlamydia.

[0250] In embodiments, the disease or disorder is induced by infection with P. gingivalis.

[0251] In embodiments, the disease or disorder is acne.

[0252] In embodiments, a method for treating or preventing a disease is provided, wherein said method comprises administering to a subject in need thereof a composition of the present invention and wherein the disease is amenable to treatment or prevention by the peptide or protein encoded by the mRNA.

[0253] In embodiments, a method for treating, preventing or ameliorating a disease, disorder or infection, wherein said method comprises administering to a subject in need thereof the composition of the present invention and wherein the disease, disorder or infection is amenable to treatment or prevention by the peptide or protein encoded by the mRNA, optionally wherein the disease is (a) a protein deficiency, optionally wherein the protein deficiency affects the liver, lung, brain or muscle, (b) an autoimmune disease, (c) an infectious disease, or (d) cancer.

[0254] In embodiments, a method for inducing an immune response in a subject is provided, wherein said method comprises administering to a subject in need thereof the composition of the present invention.

[0255] In embodiments, provided herein is the use of a composition of the present invention for the manufacture of a medicament for the treatment of a disease, wherein the disease is amenable to treatment or prevention by the peptide or protein encoded by the mRNA.

[0256] In embodiments, the composition is administered intranasally, intravenously, intrathecally or intramuscularly, or by pulmonary delivery, optionally through nebulization.

[0257] In embodiments, the composition is administered intravenously, intrathecally, intramuscularly, intranasally, sublingually, or by pulmonary delivery, optionally through nebulization. In one embodiment, the composition is administered intramuscularly. In one embodiment, the composition is administered intravenously.Exemplary Compounds

[0258] In embodiments, the cationic lipids of the present invention include compounds selected from those depicted in Table 1, or a pharmaceutically acceptable salt thereof.Table 1# CompoundkA ^^ ^ ^-MOHOH A ^z\x vH0A / Wx=z^x^ ■'X / Z^-ZXSZ\ZXXA'OHOOHA ~ x OH ( '■' ^'' x ZX zx xx x--. A xx. x~x Sx xx xX N- "■'■ ■'-" 'i o r ■*si A / ^Ayx XzH0Vvx-. / XZ ", X- ~ X~X xlHOH OHo.yxW xxK- X. X x-~X -v / x^x XOHHOH,-^ / X ^ ^•^•OHr.'Sx^-'\ZAVAX X *-.--' >< X--. N X X- •■*- / X x-*- X-. ---' xX X V N N 0’s’'xHO-" '"-^ ■'*■■' ■'■•■'■ -^' "^'HOHOHX X--- XX X— • XX x^\ -XX X-X' - X x xOH0X ^N.xX... x.... Sxg^, ZX x-"\.., - N 'I' X X A X-X. Ki. j XZx -x X X X- X x^ X x-X x x~x x x-X. xX- x.x JHOHOH>x.x-x^x^x^' / XZ^KZX / X / VOH QNx-S's^Xz^xz N xL N x" Z-x ''-•' X... 0 JL O / XXN- x^ HO A / '. ■'-■z X.. xX'x x. xx'-OHOHOH |xX xx,.,x xx.. xx "•■ / X x'x / X x'x A / X / X / X _, SX / x / .-x N■ ■ ■ ' 'i o r?3■ 'irN— O -V --... N.. X HOAX^ ZX / X^X X x''\x''Xx'Xx'Xxy0HOH" ZX ” / 'X / \ Z-X " / X. AZOH 0 < N / --X '■ zS. Sr / \ / -X - >xx>< NZx O A 0 / X N.'"" J HO Jx "- x' -X XX x=, / xx ^.-x x^ XX ^-xxx^_^x x-Xx-x / -x JOHOHOHj^xXv / x / S / x / x^x / x^x>xo / v^A^-AX N. XX A -N,-X X N. x- J"H0- A x- -X X.- -x XX XX X • H■'xX^ ^x^xx^QHOH OHrAxx. x....... - -.-..'-... Ai0^" XzX^^XXxXrN..'-XN.., N,„J HOA^XX / X s X XX.- X,. X. XX I -0HHOH OH'XXXX-xA^ 0 / x.-X^S-s-XXxNs HOAXXXX. XS^ / X ^Xx / x^.x x^ x-x ^x-x.. / > |...oi_| HpHO XX XX XX?..--N-.... S.s.. x^x, AXXX^-^xx XxZX. x x JI N 1 X XV.,vXX x- o O J. BH0',xxxx. X xx it rY °xxAOH / w xo"'^O O NS'SNorN ANN JH 0•■: H>)OHJ > o OHyV V J^> A~ T ^A 0 A NS'S' N ">-' J „ 0 A — JC^A HO 1 -■,- Y 'cr -~- J- 'OHH( OHOH ^AXA / VAX / zN, N N J > Ar JT^ HO^— — cnI OH O Z —,OH< <.SX A JJ - 6OH'' xx -- X x" x xx.> xx -y „0H Q f xxfcN^XXx. S. ^ S -x xxXfasL N.. fa' A.. fa XX J HO.1. X.. Xxx.x x^X - x=.x.- x.,.x xxx. AXx_. x„ JHOHOH XXXXxX^ oN\ 'XzxAx' |:r(X i H\XXXZXXXXOHOH( " "" ' -- xxv- XxX, -x X..,^sv, OH o r ^ xtfa yxv- X. Ss^ s, X - „ -x N \< N X X. N ’ X N s' '"•' N -- J H0 1 — " x-x xx•'XT77T. X.x-XX... -XX- X, s''' - H H OHOH OHx.. • / xA Q X 'r*^NAN.~^M,y•• • •. -. •...... •. • ••. • • •• H H'. ^.. C-HOH OH Z'.^''"'. / '-x''...z>XXXXx-XX^ o ^Nxx^S,s,^^xN'. X'xxX|MAN^^, N^xJ HO^X^XXXX^XXXX 1 H H-^X^xx''x^x-^^XX<'OHOH OH fX\ 1 xXxX^xx^Xzxo / v^ 8.yxxxXf N — N A N^X. -x N '■xJ-HO- 1- '. / / xv xzzx'-x x'x''. / - 1 H Hx / x / xxxzx^onOHOH W X / V-Z,o rN^S,s^.^., H{ ( / / H H / 4 J? o z —sOH / t OH <•. • • •.. •■•.„ ■.. ■. - ’ N 5 ' • •• ” • '*co1( o c HO^^-?oo —?zz \ Z—Q> OHOH \ <2TT,-^'NZVVZ^XZS’S-ZSXzZSXzK|A °■' >. -. -.x- zVxA-.x> o / • MU - ■• - - „„,, ^ '1.,zxX. x?CrtT \ 0)O \Ar () OHOH oV V / o~! rs / / T>> >\n(OH O\ IC? H:>X / ■'■— 21’- 'X x'Xv.-x A •'j N - '• S '- x'V ■ A. >p S 'j’'xx xx zx x.. N J H (O A-- x'x ■'•' ~ xx xx XX '- xx ' - wA / * \z '•QH X ())x z- OHz— ■OHr-N^S.s^,x, N-? Vx <^vx,«Ju / p z—o / W- OH,.x,x x-x - - x- - OH f • •x''' ■ " '■-'' ' • ' ■. x-,-X XX x-,-X X-. zx XX A(X¥XX XX S 'Sz X- XX XX -XX N XAx X X X X ••• ■ • '■< AH0.^xx^x-^.-'-x_x-XX---Xx---X.x'X. '■ 'X ■'' -'X ' ' Xx'-'- X-x " X.x "--OMnOHxjx XX. XX Y" X.. XX.. S, X. XX-X X-XVXX XX / X Zx O f 'N' Ax' 'S- X,x X^z X"■ ■"', N. X-X X-X HN’ A " S' XXx'-' N '■'■■ xi HO' Jx XX XX XX X- A. X XzX XX-OH?H- x X S X X lilxx X „>x0<N' •• '■ 'S' '-x X' 0^''■'x'-X^x^gxX.^Ax-'HOxXxX'x.„xX^x''X=.x'Xx.x'X^xX.^,.''x fH'X.x X ''-X X-'x, x"x„.xX0M< i < < 5 >j J\ \ / °oZ “OH> >o— -x- \ / \ / X o 7< o 2 / - — —OH X 7 c Ji / TO Z— 1 <9 wI / bl ' " ' 'S ^'N ' ■ H \OA ('' ■'X'''x-'x-'-x— '' • • ' xxx„ -. f"-■ OHxzG>o IZxz G>wKO=Q Q / ^z>X \ / O zZ~ v>w(A o — ) / c> / OH) / X■ \ X / X\ X / X\ XZ x~x XX x\ ^OH Q / x '-.'. N ^''x, S. xx xx \, \ xx ^-NO z~~\ XL A o z — \N 0 N HO \ / yo—_ / / 1i s OH9HOH \ \ \ / x, / ~x ^-x A^ / x. / .-s-.^x / x,. N,'"' > O |N’Sf..■N, '-.^.. N A N -^x '■-^' fK X / J HO A - ^ ^ / ^ '-^■' X X^ / x^ ]'■V.zZ'V. / '''. / ''-, / '''.,-''''OHH HOH\ XX X- 'X, / X- x-. xX*xX^ AxxoX xXxN. XX X^x XXx^..-S. XX X^ Xx"x,X _bk XX XX x^ A xNXx xx^x -N. x / JH(y A xX X\ X^xX*x x^ Xxx X 'Xx^X_XXxx^XxX\X 1 xQHHo—__ / / / x 7O 2 \ —\ T OHOHX zx X X Xx zX -X xX XX x^kX < zx N xx Xx z§s ^sz^X\x x"x\x / ■ X-.xZN. xX Xx. X XX X. / z'-\, z Jx x. X J tn x^. x-x xX XX X / HC< -X- \Z \z n tOHo / xX XX z-x ^X x~x XX X 'z ^x x~x Xx z-OH 0 X XX vN^ XX \x z'S ' zgx XX Xz zzx Xx XL N Z-X. XX z ^ -*Ck=N zx N. Jzz 0 HO’ A X' -Z XX. XX ~ X xx^x*HOHXy O Z - / \ X / o < f— — ' OHOH ^-x^'xXxxX\x'' X x-.. xx XX XXXX xXXz x^ Q xX x zSxS xx x x- x x ^Nz x.^| H H'xx^X'^Xz--^XX'Xx^0H

[0259] Any of the compounds (1, 2 and 4-51) identified in Table 1 above may be provided in the form of a pharmaceutically acceptable salt and such salts are intended to be encompassed by the present invention.

[0260] The compounds of the invention as described herein can be prepared according to methods known in the art, including the exemplary syntheses of the Examples provided herein.Nucleic Acids

[0261] The compounds of the invention as described herein can be used to prepare compositions useful for the delivery of nucleic acids.Synthesis of Nucleic Acids

[0262] Nucleic acids according to the present invention may be synthesized according to any known methods. For example, mRNAs according to the present invention may be synthesized via in vitro transcription (IVT). Briefly, IVT is typically performed with a linear or circular DNA template containing a promoter, a pool of ribonucleotide triphosphates, a buffer system that may include DTT and magnesium ions, and an appropriate RNA polymerase (e.g., T3, T7, mutated T7 or SP6 RNA polymerase), DNAse I, pyrophosphatase, and / or RNAse inhibitor. The exact conditions will vary according to the specific application.

[0263] In some embodiments, for the preparation of mRNA according to the invention, a DNA template is transcribed in vitro. A suitable DNA template typically has a promoter, for example a T3, T7, mutated T7 or SP6 promoter, for in vitro transcription, followed by desired nucleotide sequence for desired mRNA and a termination signal.

[0264] Desired mRNA sequence(s) according to the invention may be determined and incorporated into a DNA template using standard methods. For example, starting from a desired amino acid sequence (e.g., an enzyme sequence), a virtual reverse translation is carried out based on the degenerated genetic code. Optimization algorithms may then be used for selection of suitable codons. Typically, the G / C content can be optimized to achieve the highest possible G / C content on one hand, taking into the best possible account the frequency of the tRNAs according to codon usage on the other hand. The optimized RNA sequence can be established and displayed, for example, with the aid of an appropriate display device and compared with the original (wild-type) sequence. A secondary structure can also be analyzed to calculate stabilizing and destabilizing properties or, respectively, regions of the RNA.Modified mRNA

[0265] In some embodiments, mRNA according to the present invention may be synthesized as unmodified or modified mRNA. Modified mRNA comprises nucleotide modifications in the RNA. A modified mRNA according to the invention can thus include nucleotide modification that are, for example, backbone modifications, sugar modifications or base modifications. In some embodiments, mRNAs may be synthesized from naturally occurring nucleotides and / or nucleotide analogues (modified nucleotides) including, but not limited to, purines (adenine (A), guanine (G)) or pyrimidines (thymine (T), cytosine (C), uracil (U)), and as modified nucleotides analogues or derivatives of purines and pyrimidines, such as e.g., 1-methyl-adenine, 2-methyl-adenine, 2-methylthio-N-6-isopentenyl-adenine, N-6-methyl-adenine, N-6-isopentenyl-adenine, 2-thio-cytosine, 3-methyl-cytosine, 4-acetyl-cytosine, 5-methyl-cytosine, 2,6-diaminopurine, 1-methyl-guanine, 2-methyl-guanine, 2,2-dimethyl-guanine, 7-methyl-guanine, inosine, 1-methyl-inosine, pseudouracil (5-uracil), dihydro-uracil, 2-thio-uracil, 4-thio-uracil, 5-carboxymethylaminomethyl-2-thio-uracil, 5-(carboxyhydroxymethyl)-uracil, 5-fluoro-uracil, 5-bromo-uracil, 5-carboxymethylaminomethyl-uracil, 5-methyl-2-thio-uracil, 5-methyl-uracil, N-uracil-5-oxyacetic acid methyl ester, 5-methylaminomethyl-uracil, 5-methoxyaminomethyl-2-thio-uracil, 5'-methoxycarbonylmethyl-uracil, 5-methoxy-uracil, uracil-5-oxyacetic acid methyl ester, uracil-5-oxyacetic acid (v), 1-methyl-pseudouracil, queuosine, beta-D-mannosyl-queuosine, wybutoxosine, and phosphoramidates, phosphorothioates, peptide nucleotides, methylphosphonates, 7-deazaguanosine, 5-methylcytosine and inosine. The preparation of such analogues is known to a person skilled in the art e.g., from the U. S. Pat. No. 4,373,071, U. S. Pat. No. 4,401,796, U. S. Pat. No. 4,415,732, U. S. Pat. No. 4,458,066, U. S. Pat. No. 4,500,707, U. S. Pat. No. 4,668,777, U. S. Pat. No. 4,973,679, U. S. Pat. No. 5,047,524, U. S. Pat. No. 5,132,418, U. S. Pat. No. 5,153,319, U. S. Pat. Nos. 5,262,530 and 5,700,642, the disclosures of which are incorporated by reference in their entirety.Pharmaceutical Formulations of Cationic Lipids and Nucleic Acids

[0266] In certain embodiments, the compounds of the invention as described herein, as well as pharmaceutical and liposomal compositions comprising such lipids, can be used in formulations to facilitate the delivery of encapsulated materials (e.g., one or more polynucleotides such as mRNA) to, and subsequent transfection of one or more target cells. For example, in certain embodiments cationic lipids described herein (and compositions such as liposomal compositions comprising such lipids) are characterized as resulting in one or more of receptor-mediated endocytosis, clathrin-mediated and caveolae-mediated endocytosis, phagocytosis and macropinocytosis, fusogenicity,endosomal or lysosomal disruption and / or releasable properties that afford such compounds advantages relative other similarly classified lipids.

[0267] According to the present invention, a nucleic acid, e.g., mRNA encoding a protein (e.g., a full length, fragment or portion of a protein) as described herein may be delivered via a delivery vehicle comprising a compound of the invention as described herein.

[0268] As used herein, the terms "delivery vehicle," "transfer vehicle," "nanoparticle," or grammatical equivalents thereof, are used interchangeably.

[0269] For example, the present invention provides a composition (e.g., a pharmaceutical composition) comprising a cationic lipids as described herein and one or more polynucleotides. A composition (e.g., a pharmaceutical composition) may further comprise(i) one or more additional cationic lipids,(ii) one or more non-cationic lipids,(iii) one or more cholesterol-based lipids and / or(iv) one or more PEG-modified lipids.

[0270] In certain embodiments a composition exhibits an enhanced (e.g., increased) ability to transfect one or more target cells. Accordingly, also provided herein are methods of transfecting one or more target cells. Such methods generally comprise the step of contacting the one or more target cells with the cationic lipids and / or pharmaceutical compositions disclosed herein (e.g., a liposomal formulation comprising a compound described herein encapsulating one or more polynucleotides) such that the one or more target cells are transfected with the materials encapsulated therein (e.g., one or more polynucleotides). As used herein, the terms "transfect" or "transfection" refer to the intracellular introduction of one or more encapsulated materials (e.g., nucleic acids and / or polynucleotides) into a cell (e.g., into a target cell). The introduced polynucleotide may be stably or transiently maintained in the target cell. The term "transfection efficiency" refers to the relative amount of such encapsulated material (e.g., polynucleotides) up-taken by, introduced into, and / or expressed by the target cell which is subject to transfection. In practice, transfection efficiency may be estimated by the amount of a reporter polynucleotide product produced by the target cells following transfection. In certain embodiments, the compounds and pharmaceutical compositions described herein demonstrate high transfection efficiencies thereby improving the likelihood that appropriate dosages of the encapsulated materials (e.g., one or more polynucleotides) will be delivered to the site of pathology and subsequently expressed, while at the same time minimizing potential systemic adverse effects or toxicity associated with the compound or their encapsulated contents.

[0271] Following transfection of one or more target cells by, for example, the polynucleotides encapsulated in the one or more lipid nanoparticles comprising the pharmaceutical or liposomal compositions disclosed herein, the production of the product (e.g., a polypeptide or protein) encoded by such polynucleotide may be stimulated and the capability of such target cells to express the polynucleotide and produce, for example, a polypeptide or protein of interest is enhanced. For example, transfection of a target cell by one or more compounds or pharmaceutical compositions encapsulating mRNA will enhance ( / .e., increase) the production of the protein or enzyme encoded by such mRNA.

[0272] Further, delivery vehicles described herein (e.g., liposomal delivery vehicles) may be prepared to preferentially distribute to other target tissues, cells or organs, such as the heart, lungs, kidneys, spleen. In embodiments, the delivery vehicles described herein (e.g., liposomal delivery vehicles) may be prepared to preferentially distribute to the lungs. In embodiments, the lipid nanoparticles of the present invention may be prepared to achieve enhanced delivery to the target cells and tissues. For example, polynucleotides (e.g., mRNA) encapsulated in one or more of the compounds or pharmaceutical and liposomal compositions described herein can be delivered to and / or transfect targeted cells or tissues. In some embodiments, the encapsulated polynucleotides (e.g., mRNA) are capable of being expressed and functional polypeptide products produced (and in some instances excreted) by the target cell, thereby conferring a beneficial property to, for example the target cells or tissues. Such encapsulated polynucleotides (e.g., mRNA) may encode, for example, an antigen hormone, enzyme, receptor, polypeptide, peptide or other protein of interest.Liposomal Delivery Vehicles

[0273] In some embodiments, a composition is a suitable delivery vehicle. In embodiments, a composition is a liposomal delivery vehicle, e.g., a lipid nanoparticle.

[0274] The terms "liposomal delivery vehicle" and "liposomal composition" are used interchangeably.

[0275] Enriching liposomal compositions with one or more of the cationic lipids disclosed herein may be used as a means of improving the safety profile or otherwise conferring one or more desired properties to such enriched liposomal composition (e.g., improved delivery of the encapsulated polynucleotides to one or more target cells and / or reduced in vivo toxicity of a liposomal composition). Accordingly, also contemplated are pharmaceutical compositions, and in particular liposomal compositions, that comprise one or more of the cationic lipids disclosed herein.

[0276] Thus, in certain embodiments, the compounds of the invention as described herein may be used as a component of a liposomal composition to facilitate or enhance the delivery and release ofencapsulated materials (e.g., one or more therapeutic agents) to one or more target cells (e.g., by permeating or fusing with the lipid membranes of such target cells).

[0277] As used herein, liposomal delivery vehicles, e.g., lipid nanoparticles, are usually characterized as microscopic vesicles having an interior aqua space sequestered from an outer medium by a membrane of one or more bilayers. Bilayer membranes of liposomes are typically formed by amphiphilic molecules, such as lipids of synthetic or natural origin that comprise spatially separated hydrophilic and hydrophobic domains (Lasic, Trends Biotechnol., 16: 307-321, 1998). Bilayer membranes of the liposomes can also be formed by amphophilic polymers and surfactants (e.g., polymerosomes, niosomes, etc.). In the context of the present invention, a liposomal delivery vehicle typically serves to transport a desired mRNA to a target cell or tissue.

[0278] In certain embodiments, such compositions (e.g., liposomal compositions) are loaded with or otherwise encapsulate materials, such as for example, one or more biologically-active polynucleotides (e.g., mRNA).

[0279] In embodiments, a composition (e.g., a pharmaceutical composition) comprises an mRNA encoding a peptide or protein, encapsulated within a liposome. In embodiments, a liposome comprises:(i) one or more cationic lipids,(ii) one or more non-cationic lipids,(iii) one or more cholesterol-based lipids and(iv) one or more PEG-modified lipids, wherein at least one cationic lipid is a compound of the invention as described herein.

[0280] In embodiments, a composition comprises an mRNA encoding for a peptide or protein (e.g., any peptide or protein described herein). In embodiments, a composition comprises an mRNA encoding for a peptide (e.g., any peptide described herein). In embodiments, a composition comprises an mRNA encoding for a protein (e.g., any protein described herein).

[0281] In embodiments, a composition (e.g., a pharmaceutical composition) comprises a nucleic acid encapsulated within a liposome, wherein the liposome comprises a compound described herein.

[0282] In embodiments, a nucleic acid is an mRNA encoding a peptide or protein. In embodiments, an mRNA encodes a peptide or protein for use in the delivery to or treatment of the human body. Exemplary mRNAs are described herein.

[0283] In embodiments, a liposomal delivery vehicle (e.g., a lipid nanoparticle) can have a net positive charge.

[0284] In embodiments, a liposomal delivery vehicle (e.g., a lipid nanoparticle) can have a net negative charge.

[0285] In embodiments, a liposomal delivery vehicle (e.g., a lipid nanoparticle) can have a net neutral charge.

[0286] In embodiments, a lipid nanoparticle that encapsulates a nucleic acid (e.g., mRNA encoding a peptide or protein) comprises one or more compounds of the invention as described herein.

[0287] For example, the amount of a compound of the invention as described herein in a composition can be described as a percentage ("wt%") of the combined dry weight of all lipids of a composition (e.g., the combined dry weight of all lipids present in a liposomal composition).

[0288] In embodiments of the pharmaceutical compositions described herein, a compound of the invention as described herein is present in an amount that is about 0.5 wt% to about 30 wt% (e.g., about 0.5 wt% to about 20 wt%) of the combined dry weight of all lipids present in a composition (e.g., a liposomal composition).

[0289] In embodiments, a compound of the invention as described herein is present in an amount that is about 1 wt% to about 30 wt%, about 1 wt% to about 20 wt%, about 1 wt% to about 15 wt%, about 1 wt% to about 10 wt%, or about 5 wt% to about 25 wt% of the combined dry weight of all lipids present in a composition (e.g., a liposomal composition). In embodiments, a compound of the invention as described herein is present in an amount that is about 0.5 wt% to about 5 wt%, about 1 wt% to about 10 wt%, about 5 wt% to about 20 wt%, or about 10 wt% to about 20 wt% of the combined dry weight of all lipids present in a composition such as a liposomal delivery vehicle.

[0290] In embodiments, the amount of a compound of the invention as described herein is present in an amount that is at least about 5 wt%, about 10 wt%, about 15 wt%, about 20 wt%, about 25 wt%, about 30 wt%, about 35 wt%, about 40 wt%, about 45 wt%, about 50 wt%, about 55 wt%, about 60 wt%, about 65 wt%, about 70 wt%, about 75 wt%, about 80 wt%, about 85 wt%, about 90 wt%, about 95 wt%, about 96 wt%, about 97 wt%, about 98 wt%, or about 99 wt% of the combined dry weight of total lipids in a composition (e.g., a liposomal composition).

[0291] In embodiments, the amount of a compound of the invention as described herein is present in an amount that is no more than about 5 wt%, about 10 wt%, about 15 wt%, about 20 wt%, about 25 wt%, about 30 wt%, about 35 wt%, about 40 wt%, about 45 wt%, about 50 wt%, about 55 wt%, about 60 wt%, about 65 wt%, about 70 wt%, about 75 wt%, about 80 wt%, about 85 wt%, about 90 wt%, about 95 wt%, about 96 wt%, about 97 wt%, about 98 wt%, or about 99 wt% of the combined dry weight of total lipids in a composition (e.g., a liposomal composition).

[0292] In embodiments, a composition (e.g., a liposomal delivery vehicle such as a lipid nanoparticle) comprises about 0.1 wt% to about 20 wt% (e.g., about 0.1 wt% to about 15 wt%) of acompound described herein. In embodiments, a delivery vehicle (e.g., a liposomal delivery vehicle such as a lipid nanoparticle) comprises about 0.5 wt%, about 1 wt%, about 3 wt%, about 5 wt%, or about 10 wt% of a compound described herein. In embodiments, a delivery vehicle (e.g., a liposomal delivery vehicle such as a lipid nanoparticle) comprises up to about 0.5 wt%, about 1 wt%, about 3 wt%, about 5 wt%, about 10 wt%, about 15 wt%, or about 20 wt% of a compound described herein. In embodiments, the percentage results in an improved beneficial effect (e.g., improved delivery to targeted tissues).

[0293] The amount of a compound of the invention as described herein in a composition also can be described as a percentage ("mol%") of the combined molar amounts of total lipids of a composition (e.g., the combined molar amounts of all lipids present in a liposomal delivery vehicle).

[0294] In embodiments of pharmaceutical compositions described herein, a compound of the invention as described herein is present in an amount that is about 0.5 mol% to about 50 mol% (e.g., about 0.5 mol% to about 20 mol%) of the combined molar amounts of all lipids present in a composition such as a liposomal delivery vehicle.

[0295] In embodiments, a compound of the invention as described herein is present in an amount that is about 0.5 mol% to about 5 mol%, about 1 mol% to about 10 mol%, about 5 mol% to about 20 mol%, about 10 mol% to about 20 mol%, about 15 mol% to about 30 mol%, about 20 mol% to about 35 mol%, about 25 mol% to about 40 mol%, about 30 mol% to about 45 mol%, about 35 mol% to about 50 mol%, about 40 mol% to about 55 mol %, or about 45 mol% to about 60 mol% of the combined molar amounts of all lipids present in a composition such as a liposomal delivery vehicle. In embodiments, a compound of the invention as described herein is present in an amount that is about 1 mol% to about 60 mol%, 1 mol% to about 50 mol%, 1 mol% to about 40 mol%, 1 mol% to about 30 mol%, about 1 mol% to about 20 mol%, about 1 mol% to about 15 mol%, about 1 mol% to about 10 mol%, about 5 mol% to about 55 mol%, about 5 mol% to about 45 mol%, about 5 mol% to about 35 mol% or about 5 mol% to about 25 mol% of the combined molar amounts of all lipids present in a composition such as a liposomal delivery vehicle.

[0296] In certain embodiments, a compound of the invention as described herein can comprise from about 0.1 mol% to about 50 mol%, or from 0.5 mol% to about 50 mol%, or from about 1 mol% to about 50 mol%, or from about 5 mol% to about 50 mol%, or from about 10 mol% to about 50 mol%, or from about 15 mol% to about 50 mol%, or from about 20 mol% to about 50 mol%, or from about 25 mol% to about 50 mol%, or from about 30 mol% to about 50 mol%, of the total amount of lipids in a composition (e.g., a liposomal delivery vehicle).

[0297] In certain embodiments, a compound of the invention as described herein can comprise greater than about 0.1 mol%, or greater than about 0.5 mol%, or greater than about 1 mol%, greaterthan about 5 mol%, greater than about 10 mol%, greater than about 20 mol%, greater than about 30 mol%, or greater than about 40 mol% of the total amount of lipids in the lipid nanoparticle.

[0298] In certain embodiments, a compound as described can comprise less than about 60 mol%, or less than about 55 mol%, or less than about 50 mol%, or less than about 45 mol%, or less than about 40 mol%, or less than about 35 mol %, less than about 30 mol%, or less than about 25 mol%, or less than about 10 mol%, or less than about 5 mol%, or less than about 1 mol% of the total amount of lipids in a composition (e.g., a liposomal delivery vehicle).

[0299] In embodiments, the amount of a compound of the invention as described herein is present in an amount that is at least about 5 mol%, about 10 mol%, about 15 mol%, about 20 mol%, about 25 mol%, about 30 mol%, about 35 mol%, about 40 mol%, about 45 mol%, about 50 mol%, about 55 mol%, about 60 mol%, about 65 mol%, about 70 mol%, about 75 mol%, about 80 mol%, about 85 mol%, about 90 mol%, about 95 mol%, about 96 mol%, about 97 mol%, about 98 mol%, or about 99 mol% of the combined molar amounts of total lipids in a composition (e.g., a liposomal composition).

[0300] In embodiments, the amount of a compound of the invention as described herein is present in an amount that is no more than about 5 mol%, about 10 mol%, about 15 mol%, about 20 mol%, about 25 mol%, about 30 mol%, about 35 mol%, about 40 mol%, about 45 mol%, about 50 mol%, about 55 mol%, about 60 mol%, about 65 mol%, about 70 mol%, about 75 mol%, about 80 mol%, about 85 mol%, about 90 mol%, about 95 mol%, about 96 mol%, about 97 mol%, about 98 mol%, or about 99 mol% of the combined molar amounts of total lipids in a composition (e.g., a liposomal composition).

[0301] In embodiments, the percentage results in an improved beneficial effect (e.g., improved delivery to targeted tissues).

[0302] In a typical embodiment, a composition of the invention (e.g., a liposomal composition) comprises:(i) one or more cationic lipids,(ii) one or more non-cationic lipids,(iii) one or more cholesterol-based lipids, and(iv) one or more PEG-modified lipids, wherein at least one cationic lipid is a compound of the invention as described herein.

[0303] For example, a composition suitable for practicing the invention has four lipid components comprising a compound of the invention as described herein as the cationic lipid component, and further comprising:a non-cationic lipid,(ii) a cholesterol-based lipid and(iii) a PEG-modified lipid.

[0304] The non-cationic lipid may be DOPE or DEPE. The cholesterol-based lipid may be cholesterol. The PEG-modified lipid may be DMG-PEG2K.

[0305] In further embodiments, pharmaceutical (e.g., liposomal) compositions comprise one or more of a PEG-modified lipid, a non-cationic lipid and a cholesterol lipid. In other embodiments, such pharmaceutical (e.g., liposomal) compositions comprise: one or more PEG-modified lipids; one or more non-cationic lipids; and one or more cholesterol lipids. In yet further embodiments, such pharmaceutical (e.g., liposomal) compositions comprise: one or more PEG-modified lipids and one or more cholesterol lipids.

[0306] In embodiments, a composition (e.g., lipid nanoparticle) that encapsulates a nucleic acid (e.g., mRNA encoding a peptide or protein) comprises one or more compounds of the invention as described herein, and one or more lipids selected from the group consisting of a cationic lipid, a noncationic lipid, and a PEGylated lipid.

[0307] In embodiments, a composition (e.g., lipid nanoparticle) that encapsulates a nucleic acid (e.g., mRNA encoding a peptide or protein) comprises one or more compound of the invention as described herein; one or more lipids selected from the group consisting of a cationic lipid, a noncationic lipid, and a PEGylated lipid; and further comprises a cholesterol-based lipid. Typically, such a composition has four lipid components comprising a compound of the invention as described herein as the cationic lipid component, and further comprising:(i) a non-cationic lipid (e.g., DOPE),(ii) a cholesterol-based lipid (e.g., cholesterol) and(iii) a PEG-modified lipid (e.g., DMG-PEG2K).

[0308] In embodiments, a lipid nanoparticle that encapsulates a nucleic acid (e.g., mRNA encoding a peptide or protein) comprises one or more compounds of the invention as described herein, as well as one or more lipids selected from the group consisting of:(i) a cationic lipid,(ii) a non-cationic lipid,(iii) a PEGylated lipid, and(iv) a cholesterol-based lipid.

[0309] According to various embodiments, the selection of cationic lipids, non-cationic lipids and / or PEG-modified lipids which comprise the lipid nanoparticle, as well as the relative molar ratio of such lipids to each other, is based upon the characteristics of the selected lipid(s), the nature of the intended target cells, the characteristics of the mRNA to be delivered. Additional considerationsinclude, for example, the saturation of the alkyl chain, as well as the size, charge, pH, pKa, fusogenicity and toxicity of the selected lipid(s). Thus, the molar ratios may be adjusted accordingly.Cationic Lipids

[0310] In addition to any of the compounds of the invention as described herein, a composition may comprise one or more additional cationic lipids.

[0311] In some embodiments, liposomes may comprise one or more additional cationic lipids. As used herein, the phrase "cationic lipid" refers to any of a number of lipid species that have a net positive charge at a selected pH, such as physiological pH. Several cationic lipids have been described in the literature, many of which are commercially available.

[0312] Suitable additional cationic lipids for use in the compositions include the cationic lipids as described in the literature.Helper Lipids

[0313] Compositions (e.g., liposomal compositions) may also comprise one or more helper lipids. Such helper lipids include non-cationic lipids. As used herein, the phrase "non-cationic lipid" refers to any neutral, zwitterionic or anionic lipid. As used herein, the phrase "anionic lipid" refers to any of a number of lipid species that carry a net negative charge at a selected pH, such as physiological pH. Non-cationic lipids include, but are not limited to, distearoylphosphatidylcholine (DSPC), dioleoylphosphatidylcholine (DOPC), dipalmitoylphosphatidylcholine (DPPC), dioleoylphosphatidylglycerol (DOPG), dipalmitoylphosphatidylglycerol (DPPG), dioleoylphosphatidylethanolamine (DOPE), l,2-Dierucoyl-sn-glycero-3-phosphoethanolamine (DEPE), palmitoyloleoylphosphatidylcholine (POPC), palmitoyloleoyl-phosphatidylethanolamine (POPE), dioleoyl-phosphatidylethanolamine 4-(N-maleimidomethyl)-cyclohexane-l-carboxylate (DOPE-mal), dipalmitoyl phosphatidyl ethanolamine (DPPE), dimyristoylphosphoethanolamine (DMPE), distearoyl-phosphatidyl-ethanolamine (DSPE), 16-O-monomethyl PE, 16-O-dimethyl PE, 18-1-trans PE, l-stearoyl-2-oleoyl-phosphatidyethanolamine (SOPE), or a mixture thereof. A noncationic or helper lipid suitable for practicing the invention is dioleoylphosphatidylethanolamine (DOPE). Alternatively, l,2-Dierucoyl-sn-glycero-3-phosphoethanolamine (DEPE) can be used as a non-cationic or helper lipid.

[0314] In some embodiments, a non-cationic lipid is a neutral lipid, i.e., a lipid that does not carry a net charge in the conditions under which the composition is formulated and / or administered.

[0315] In some embodiments, a non-cationic lipid may be present in a molar ratio (mol%) of about 5% to about 90%, about 5% to about 70%, about 5% to about 50%, about 5% to about 40%, about5% to about 30%, about 10% to about 70%, about 10% to about 50%, or about 10% to about 40% of the total lipids present in a composition. In some embodiments, total non-cationic lipids may be present in a molar ratio (mol%) of about 5% to about 90%, about 5% to about 70%, about 5% to about 50%, about 5% to about 40%, about 5% to about 30%, about 10 % to about 70%, about 10% to about 50%, or about 10% to about 40% of the total lipids present in a composition. In some embodiments, the percentage of non-cationic lipid in a liposome may be greater than about 5 mol%, greater than about 10 mol%, greater than about 20 mol%, greater than about 30 mol%, or greater than about 40 mol%. In some embodiments, the percentage total non-cationic lipids in a liposome may be greater than about 5 mol%, greater than about 10 mol%, greater than about 20 mol%, greater than about 30 mol%, or greater than about 40 mol%. In some embodiments, the percentage of non-cationic lipid in a liposome is no more than about 5 mol%, no more than about 10 mol%, no more than about 20 mol%, no more than about 30 mol%, or no more than about 40 mol%. In some embodiments, the percentage total non-cationic lipids in a liposome may be no more than about 5 mol%, no more than about 10 mol%, no more than about 20 mol%, no more than about 30 mol%, or no more than about 40 mol%.

[0316] In some embodiments, a non-cationic lipid may be present in a weight ratio (wt%) of about 5% to about 90%, about 5% to about 70%, about 5% to about 50%, about 5% to about 40%, about 5% to about 30%, about 10 % to about 70%, about 10% to about 50%, or about 10% to about 40% of the total lipids present in a composition. In some embodiments, total non-cationic lipids may be present in a weight ratio (wt%) of about 5% to about 90%, about 5% to about 70%, about 5% to about 50%, about 5% to about 40%, about 5% to about 30%, about 10 % to about 70%, about 10% to about 50%, or about 10% to about 40% of the total lipids present in a composition. In some embodiments, the percentage of non-cationic lipid in a liposome may be greater than about 5 wt%, greater than about 10 wt%, greater than about 20 wt%, greater than about 30 wt%, or greater than about 40 wt%. In some embodiments, the percentage total non-cationic lipids in a liposome may be greater than about 5 wt%, greater than about 10 wt%, greater than about 20 wt%, greater than about 30 wt%, or greater than about 40 wt%. In some embodiments, the percentage of non-cationic lipid in a liposome is no more than about 5 wt%, no more than about 10 wt%, no more than about 20 wt%, no more than about 30 wt%, or no more than about 40 wt%. In some embodiments, the percentage total non-cationic lipids in a liposome may be no more than about 5 wt%, no more than about 10 wt%, no more than about 20 wt%, no more than about 30 wt%, or no more than about 40 wt%.Cholesterol-based Lipids

[0317] In some embodiments, a composition (e.g., a liposomal composition) comprising a cationic lipid of the present invention further comprises one or more cholesterol-based lipids. For example, a suitable cholesterol-based lipid for practicing the invention is cholesterol. Other suitable cholesterol-based lipids include, for example, DC-Chol (N, N-dimethyl-N-ethylcarboxamidocholesterol), l,4-bis(3-N-oleylamino-propyl)piperazine (Gao, et al. Biochem. Biophys. Res. Comm. 179, 280 (1991); Wolf et al. BioTechniques 23, 139 (1997); U. S. Pat. No. 5,744,335), beta-sitosterol, or imidazole cholesterol ester (ICE), which has the following structure,

[0318] In some embodiments, a cholesterol-based lipid may be present in a molar ratio (mol%) of about 1% to about 30%, or about 5% to about 20% of the total lipids present in a liposome. In some embodiments, the percentage of cholesterol-based lipid in the lipid nanoparticle may be greater than about 5 mol%, greater than about 10 mol%, greater than about 20 mol%, greater than about 30 mol%, or greater than about 40 mol%. In some embodiments, the percentage of cholesterol-based lipid in the lipid nanoparticle may be no more than about 5 mol%, no more than about 10 mol%, no more than about 20 mol%, no more than about 30 mol%, or no more than about 40 mol%.

[0319] In some embodiments, a cholesterol-based lipid may be present in a weight ratio (wt%) of about 1% to about 30%, or about 5% to about 20% of the total lipids present in a liposome. In some embodiments, the percentage of cholesterol-based lipid in the lipid nanoparticle may be greater than about 5 wt%, greater than about 10 wt%, greater than about 20 wt%, greater than about 30 wt%, or greater than about 40 wt%. In some embodiments, the percentage of cholesterol-based lipid in the lipid nanoparticle may be no more than about 5 wt%, no more than about 10 wt%, no more than about 20 wt%, no more than about 30 wt%, or no more than about 40 wt%.PEGylated Lipids

[0320] In some embodiments, a composition (e.g., a liposomal composition) comprises one or more further PEGylated lipids. A suitable PEG-modified or PEGylated lipid for practicing the invention is l,2-dimyristoyl-rac-glycero-3-methoxypolyethylene glycol-2000 (DMG-PEG2K).

[0321] For example, the use of polyethylene glycol (PEG)-modified phospholipids and derivatized lipids such as derivatized ceramides (PEG-CER), including N-octanoyl-sphingosine-1-[succinyl(methoxy polyethylene glycol)-2000] (C8 PEG-2000 ceramide) is also contemplated by the present invention in combination with one or more of compounds of the invention as described herein and, in some embodiments, other lipids together which comprise the liposome. In some embodiments, particularly useful exchangeable lipids are PEG-ceramides having shorter acyl chains (e.g., (C14) or (C18)).

[0322] Contemplated further PEG-modified lipids (also referred to herein as a PEGylated lipid, which term is interchangeable with PEG-modified lipid) include, but are not limited to, a polyethylene glycol chain of up to 5 kDa in length covalently attached to a lipid with alkyl chain(s) of (C6-C20) length. In some embodiments, a PEG-modified or PEGylated lipid is PEGylated cholesterol or PEG-2K. The addition of such components may prevent complex aggregation and may also provide a means for increasing circulation lifetime and increasing the delivery of the lipid-nucleic acid composition to the target cell, (Klibanov et al. (1990) FEBS Letters, 268 (1): 235-237), or they may be selected to rapidly exchange out of the formulation in vivo (see U. S. Pat. No. 5,885,613).

[0323] Further PEG-modified phospholipid and derivatized lipids of the present invention may be present in a molar ratio (mol%) from about 0% to about 10%, about 0.5% to about 10%, about 1% to about 10%, about 2% to about 10%, about 3% to about 5%, about 1% to about 5%, or about 1.5% to about 3% of the total lipid present in the composition (e.g., a liposomal composition).Pharmaceutical Formulations and Therapeutic Uses

[0324] Compounds of the invention as described herein may be used in the preparation of compositions (e.g., to construct liposomal compositions) that facilitate or enhance the delivery and release of encapsulated materials (e.g., one or more therapeutic polynucleotides) to one or more target cells (e.g., by permeating or fusing with the lipid membranes of such target cells).

[0325] For example, when a liposomal composition (e.g., a lipid nanoparticle) comprises or is otherwise enriched with one or more of the compounds disclosed herein, the phase transition in the lipid bilayer of the one or more target cells may facilitate the delivery of the encapsulated materials (e.g., one or more therapeutic polynucleotides encapsulated in a lipid nanoparticle) into the one or more target cells.

[0326] Similarly, in certain embodiments compounds of the invention as described herein may be used to prepare liposomal vehicles that are characterized by their reduced toxicity in vivo. In certain embodiments, the reduced toxicity is a function of the high transfection efficiencies associated withthe compositions disclosed herein, such that a reduced quantity of such composition may be administered to the subject to achieve a desired therapeutic response or outcome.

[0327] In certain embodiments, compounds of the invention as described herein may be used to prepare liposomal vehicles that are characterized by effective intranasal delivery of mRNA. In certain embodiments, compounds of the invention as described herein may be used to prepare liposomal vehicles that are characterized by effective pulmonary delivery of mRNA. In certain embodiments, compounds of the invention as described herein may be used to prepare liposomal vehicles that are characterized by achieving high levels of peptide or protein expression when delivering mRNA encoding for said peptide or protein by intravenous, intrathecal, intramuscular, intranasal, sublingual, or by pulmonary delivery, optionally through nebulization. In certain embodiments, compounds of the invention as described herein may be used to prepare liposomal vehicles that are characterized by achieving high levels of peptide or protein expression when delivering mRNA encoding for said peptide or protein by intramuscular delivery.

[0328] Thus, pharmaceutical formulations comprising a compound described and nucleic acids provided by the present invention may be used for various therapeutic disease and / or disease prevention purposes. To facilitate delivery of nucleic acids in vivo, a compound described herein and nucleic acids can be formulated in combination with one or more additional pharmaceutical carriers, targeting ligands or stabilizing reagents. In some embodiments, a compound described herein can be formulated via pre-mixed lipid solution. In other embodiments, a composition comprising a compound described herein can be formulated using post-insertion techniques into the lipid membrane of the nanoparticles. Techniques for formulation and administration of drugs may be found in " Remington's Pharmaceutical Sciences," Mack Publishing Co., Easton, Pa., latest edition.

[0329] Suitable routes of administration include, for example, oral, rectal, vaginal, transmucosal, pulmonary including intratracheal or inhaled, or intestinal administration; parenteral delivery, including intradermal, transdermal (topical), intramuscular, subcutaneous, intramedullary injections, as well as intrathecal, direct intraventricular, intravenous, intraperitoneal, or intranasal. In embodiments, the route of administration is selected from intravenous, intrathecal, intramuscular, intranasal, sublingual, or by pulmonary delivery, optionally through nebulization. In embodiments, the route of administration can be intramuscular. In particular embodiments, the intramuscular administration is to a muscle selected from the group consisting of skeletal muscle, smooth muscle and cardiac muscle. In some embodiments the administration results in delivery of the nucleic acids to a muscle cell. In some embodiments the administration results in delivery of the nucleic acids to a hepatocyte ( / .e., liver cell).

[0330] One route for administering a liposomal composition of the invention may be intravenous delivery, in particular when treating metabolic disorders, especially those affecting the liver (e.g., ornithine transcarbamylase (OTC) deficiency). Alternatively, depending on the disease or disorder to be treated, the liposomal composition may be administered via pulmonary delivery (e.g., for the treatment of cystic fibrosis). For vaccination, a liposomal composition of the invention is typically administered intramuscularly. Alternatively, a liposomal composition of the invention may be administered intranasally for vaccination. Diseases or disorders affecting the eye may be treated by administering a liposomal composition of the invention intravitreally.

[0331] Alternatively or additionally, pharmaceutical formulations of the invention may be administered in a local rather than systemic manner, for example, via injection of the pharmaceutical formulation directly into a targeted tissue (e.g., in a sustained release formulation). Local delivery can be affected in various ways, depending on the tissue to be targeted. Exemplary tissues in which mRNA may be delivered and / or expressed include, but are not limited to the liver, kidney, heart, spleen, serum, brain, skeletal muscle, lymph nodes, skin, and / or cerebrospinal fluid. For example, aerosols containing compositions of the present invention can be inhaled (for nasal, tracheal, or bronchial delivery); compositions of the present invention can be injected into the site of injury, disease manifestation, or pain, for example; compositions can be provided in lozenges for oral, tracheal, or esophageal application; can be supplied in liquid, tablet or capsule form for administration to the stomach or intestines, can be supplied in suppository form for rectal or vaginal application; or can even be delivered to the eye by use of creams, drops, or even injection.

[0332] Alternatively or additionally, pharmaceutical formulations of the invention may be administered intranasally. For example, the pharmaceutical formulations of the invention may be administered via nasal spray. Exemplary tissues in which mRNA may be delivered and / or expressed include, but are not limited to the lungs, heart, liver, spleen and muscle. In embodiments, the tissue to be targeted is in the lungs. In embodiments, the tissue to be targeted is in muscle.

[0333] Alternatively or additionally, pharmaceutical formulations of the invention may be administered by pulmonary delivery, optionally through nebulization or dry powder inhalation. In embodiments, the pharmaceutical formulations of the invention are administered by pulmonary delivery through nebulization. In embodiments, the pharmaceutical formulations of the invention are administered by pulmonary delivery through dry powder inhalation. Exemplary tissues in which mRNA may be delivered and / or expressed include, but are not limited to the lungs, heart, liver, spleen and muscle. In embodiments, the tissue to be targeted is in the lungs. In embodiments, the tissue to be targeted is in muscle.

[0334] Compositions described herein can comprise mRNA encoding peptides including those described herein (e.g., a polypeptide such as a protein).

[0335] In embodiments, a mRNA encodes a polypeptide. In embodiments, a mRNA encodes a peptide. In embodiments, the peptide is an antigen. In embodiments, a mRNA encodes a peptide for treating or preventing Flu. In embodiments, a mRNA encodes a peptide for treating or preventing influenza A virus. In embodiments, a mRNA encodes a peptide for treating or preventing influenza B virus. In embodiments, a mRNA encodes a peptide for treating or preventing chlamydia. In embodiments, a mRNA encodes a peptide for treating or preventing a disease or disorder induced by infection with P. gingivalis. In embodiments, a mRNA encodes a peptide for treating or preventing acne.

[0336] In embodiments, a mRNA encodes a protein. In embodiments, a mRNA encodes a protein for treating or preventing Flu. In embodiments, a mRNA encodes a protein for treating or preventing influenza A virus. In embodiments, a mRNA encodes a protein for treating or preventing influenza B virus. In embodiments, a mRNA encodes a protein for treating or preventing chlamydia. In embodiments, a mRNA encodes a protein for treating or preventing a disease or disorder induced by infection with P. gingivalis. In embodiments, a mRNA encodes a protein for treating or preventing acne.

[0337] In embodiments, the mRNA encodes an influenza antigen. In embodiments, the mRNA encodes an antigen of influenza A virus. In embodiments, the mRNA encodes an antigen of influenza B virus. In embodiments, the mRNA encodes an antigen of Chlamydia sp. bacteria. In embodiments, the mRNA encodes an antigen of C.trachomatis. In embodiments, the mRNA encodes an antigen of P. gingivalis. In embodiments, the mRNA encodes an acne antigen. In embodiments, the mRNA encodes an antigen of C.acnes.

[0338] The present invention provides methods for delivering a composition having full-length mRNA molecules encoding a peptide or protein of interest for use in the treatment of a subject, e.g., a human subject or a cell of a human subject or a cell that is treated and delivered to a human subject.

[0339] In embodiments, compositions comprising a cationic lipid described herein are provided for use in treating or preventing Flu in a subject (e.g. human). In embodiments, compositions comprising a cationic lipid described herein are provided for use in treating or preventing a disease or disorder caused by influenza A virus in a subject (e.g. human). In embodiments, compositions comprising a cationic lipid described herein are provided for use in treating or preventing a disease or disorder caused by influenza B virus in a subject (e.g. human). In embodiments, compositions comprising a cationic lipid described herein are provided for use in treating or preventing a chlamydia in a subject(e.g. human). In embodiments, compositions comprising a cationic lipid described herein are provided for use in treating or preventing C.trachomatis infection in a subject (e.g. human). In embodiments, compositions comprising a cationic lipid described herein are provided for use in treating or preventing acne in a subject (e.g. human). In embodiments, compositions comprising a cationic lipid described herein are provided for use in treating or preventing C.acnes infection in a subject (e.g. human). In embodiments, compositions comprising a cationic lipid described herein are provided for use in treating or preventing a disease or disorder induced by infection with P. gingivalis. In any of these embodiments, said composition further comprises mRNA encoding a polypeptide, peptide or antigen suitable for the treatment or prevention of said disease or disorder or infection, such as those described herein.

[0340] Provided herein is a method of treating or preventing Flu in a subject (e.g. human), the method comprising administering a composition of the invention comprising a cationic lipid described herein to the subject. Provided herein is a method of treating or preventing a disease or disorder caused by influenza A virus in a subject (e.g. human), the method comprising administering a composition of the invention comprising a cationic lipid described herein to the subject. Provided herein is a method of treating or preventing a disease or disorder caused by influenza B virus in a subject (e.g. human), the method comprising administering a composition of the invention comprising a cationic lipid described herein to the subject. Provided herein is a method of treating or preventing chlamydia in a subject (e.g. human), the method comprising administering a composition of the invention comprising a cationic lipid described herein to the subject. Provided herein is a method of treating or preventing C.trachomatis infection in a subject (e.g. human), the method comprising administering a composition of the invention comprising a cationic lipid described herein to the subject. Provided herein is a method of treating or preventing acne in a subject (e.g. human), the method comprising administering a composition of the invention comprising a cationic lipid described herein to the subject. Provided herein is a method of treating or preventing C.acnes infection in a subject (e.g. human), the method comprising administering a composition of the invention comprising a cationic lipid described herein to the subject. Provided herein is a method of treating or preventing a disease or disorder induced by infection with P. gingivalis in a subject (e.g. human), the method comprising administering a composition of the invention comprising a cationic lipid described herein to the subject. In any of these embodiments, said composition further comprises mRNA encoding a polypeptide, peptide or antigen suitable for the treatment or prevention of said disease or disorder or infection, such as those described herein.Delivery Methods

[0341] The route of delivery used in the methods of the invention allows for non-invasive, selfadministration of the compounds of the invention. In some embodiments, the methods involve intranasal, intratracheal or pulmonary administration by aerosolization, nebulization, or instillation of a composition comprising mRNA encoding a therapeutic peptide or protein in a suitable transfection or lipid carrier vehicles as described above. In some embodiments, the methods involve intranasal, intratracheal or pulmonary administration by intravenous, intrathecal, intramuscular, intranasal, sublingual, or by pulmonary delivery, optionally through nebulization of a composition comprising mRNA encoding a therapeutic peptide or protein in a suitable transfection or lipid carrier vehicles as described above. In some embodiments, the administration is intramuscular. In some embodiments, the peptide or protein is encapsulated with a liposome. In some embodiments, the liposome comprises a lipid, which is a compound of the invention. As used herein below, administration of a compound of the invention includes administration of a composition comprising a compound of the invention.

[0342] In certain embodiments compositions of the invention are provided for use in skin injection, e.g., in the epidermis, the dermis or the hypodermis of the skin. In some embodiments, the compositions are provided in a device suitable for skin injection, such as a needle (e.g., an epidermic, dermic or hypodermic needle), a needle free device, a microneedle device or a microprojection array device. Examples of microneedle or microprojection array devices suitable for the skin injection as described in US20230270842A1, US20220339416A1, US20210085598A1, US20200246450A1, US20220143376A1, US20180264244A1, US20180263641A1, US20110245776A1.

[0343] Following administration of the composition to the subject, the peptide or protein product encoded by the mRNA (e.g., a functional protein or enzyme) is detectable in the peripheral target tissues for at least about one to seven days or longer following administration of the compound to the subject. The amount of peptide or protein product necessary to achieve a therapeutic effect will vary depending on the condition being treated, the peptide or protein encoded, and the condition of the patient. For example, the peptide or protein product may be detectable in the peripheral target tissues at a concentration (e.g., a therapeutic concentration) of at least 0.025-1.5 pg / ml (e.g., at least 0.050 pg / ml, at least 0.075 pg / ml, at least 0.1 pg / ml, at least 0.2 pg / ml, at least 0.3 pg / ml, at least 0.4 pg / ml, at least 0.5 pg / ml, at least 0.6 pg / ml, at least 0.7 pg / ml, at least 0.8 pg / ml, at least 0.9 pg / ml, at least 1.0 pg / ml, at least 1.1 pg / ml, at least 1.2 pg / ml, at least 1.3 pg / ml, at least 1.4 pg / ml, or at least 1.5 pg / ml), for at least about 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11,12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 35, 40, 45 days or longer following administration of the compound to the subject.

[0344] It has been demonstrated that nucleic acids can be delivered to the lungs by intratracheal administration of a liquid suspension of the compound and inhalation of an aerosol mist produced by a liquid nebulizer or the use of a dry powder apparatus such as that described in U. S. patent 5,780,014, incorporated herein by reference.

[0345] In certain embodiments, the compounds of the invention may be formulated such that they may be aerosolized or otherwise delivered as a particulate liquid or solid prior to or upon administration to the subject. Such compounds may be administered with the assistance of one or more suitable devices for administering such solid or liquid particulate compositions (such as, e.g., an aerosolized aqueous solution or suspension) to generate particles that are easily respirable or inhalable by the subject. In some embodiments, such devices (e.g., a metered dose inhaler, jetnebulizer, ultrasonic nebulizer, dry-powder-inhalers, propellant-based inhaler or an insufflator) facilitate the administration of a predetermined mass, volume or dose of the compositions (e.g., about 0.5 mg / kg of mRNA per dose) to the subject. For example, in certain embodiments, the compounds of the invention are administered to a subject using a metered dose inhaler containing a suspension or solution comprising the compound and a suitable propellant. In certain embodiments, the compounds of the invention may be formulated as a particulate powder (e.g., respirable dry particles) intended for inhalation. In certain embodiments, compositions of the invention formulated as respirable particles are appropriately sized such that they may be respirable by the subject or delivered using a suitable device (e.g., a mean D50 or D90 particle size less than about 500μm, 400μm, 300μm, 250μm, 200μm, 150μm, 100μm, 75μm, 50μm, 25μm, 20μm, 15μm, 12.5μm, 10μm, 5μm, 2.5μm or smaller). In yet other embodiments, the compounds of the invention are formulated to include one or more pulmonary surfactants (e.g., lamellar bodies). In some embodiments, the compounds of the invention are administered to a subject such that a concentration of at least 0.05 mg / kg, at least 0.1 mg / kg, at least 0.5 mg / kg, at least 1.0 mg / kg, at least 2.0 mg / kg, at least 3.0 mg / kg, at least 4.0 mg / kg, at least 5.0 mg / kg, at least 6.0 mg / kg, at least 7.0 mg / kg, at least 8.0 mg / kg, at least 9.0 mg / kg, at least 10 mg / kg, at least 15 mg / kg, at least 20 mg / kg, at least 25 mg / kg, at least 30 mg / kg, at least 35 mg / kg, at least 40 mg / kg, at least 45 mg / kg, at least 50 mg / kg, at least 55 mg / kg, at least 60 mg / kg, at least 65 mg / kg, at least 70 mg / kg, at least 75 mg / kg, at least 80 mg / kg, at least 85 mg / kg, at least 90 mg / kg, at least 95 mg / kg, or at least 100 mg / kg body weight is administered in a single dose. In some embodiments, the compounds of the invention are administered to a subject such that a total amount of at least 0.1 mg, at least 0.5 mg, at least 1.0 mg, at least 2.0 mg, at least 3.0 mg, at least 4.0 mg, at least 5.0 mg, at least 6.0 mg, at least 7.0 mg, at least 8.0 mg, at least 9.0 mg, at least 10 mg, at least 15 mg, at least 20 mg, at least 25 mg, at least 30 mg, at least 35 mg, at least 40 mg, at least 45 mg, at least 50 mg, at least 55 mg, atleast 60 mg, at least 65 mg, at least 70 mg, at least 75 mg, at least 80 mg, at least 85 mg, at least 90 mg, at least 95 mg or at least 100 mg mRNA is administered in one or more doses.EXAMPLES

[0346] While certain compounds, compositions and methods of the present invention have been described with specificity in accordance with certain embodiments, the following examples serve only to illustrate the compounds of the invention and are not intended to limit the same.List of abbreviations:ACN: AcetonitrileCDI: l,l'-Carbonyldiimidazole (also referred to as bis(l-imidazolyl)methanone)DCM: DichloromethaneDIPEA: N, N-DiisopropylethylamineDMAP: 4-DimethylaminopyridineDPDS: 2-(2-pyridyldithio)pyridineEDC: l-Ethyl-3-(3-dimethylaminopropyl)carbodiimideEtOAc: Ethyl acetateHATU: Hexafluorophosphate Azabenzotriazole Tetramethyl UroniumHF: Hydrofluoric acidHCI: Hydrochloric acidIPA: Isopropyl alcoholLiOH: Lithium hydroxideMeOH: MethanolNaHCO₃: Sodium hydrogencarbonatePh₃CSH: TriphenylmethanethiolPy: PyridineNa₂SO₄: Sodium SulfateTBDMSCI: Tert-butyldimethylsilyl chlorideTES: TriethylsilaneTFA: Trifluoroacetic AcidTHF: TetrahydrofuranCAD: Charged Aerosol DetectionMS: Mass spectrometryELSD: Evaporative Light-Scattering DetectionESI-MS: Electrospray ionization mass spectrometrySM: Starting MaterialTLC: Thin Layer ChromatographyExample 1: Synthesis of Compounds of the Invention

[0347] The present invention provides cationic lipids that can be prepared from readily available starting reagents. The cores of the cationic lipids of the present invention are based on the " Good" HEPES, HEPPS, and HEPBS buffers (see Table A below). Examples of tails that can be used in the cationic lipids of the present invention are provided in Table B below. Other examples of suitable lipid tails are depicted in the description and examples herein.Table A: Examples of " Good" buffers" Good" Buffer name StructureHEPES HO.I Ji 0sxo'0HHEPPS HO.N1 o I I \\ / OHx\ 0 HEPBSOz / 0HTable B: Examples of lipid chains that are suitable for the present invention at positions RPage 97 of 184Page 98 of 184Example 1A. Synthesis of Compound 1

[0348] For example, Compound 1 may be prepared according to Scheme 1 (as depicted in Fig. 1).Intermediate [31

[0349] As depicted in Scheme 1: To a stirred solution of 4-amino-2-methylbutanoic acid hydrochloride (1.2 g, 7.86 mmol, 1.0 eq) in methanol (20 ml), 1,2-epoxydecane (4.35 g, 23.5 mmol, 3.0 eq) was added followed by addition of DIPEA (3.05g, 23.5 mmol, 3.0 eq). The resulting reaction mixture was heated to 90°C for 16 h. Progress of reaction was monitored by ELSD / TLC (SM consumed). The resulting reaction mixture was cooled to room temperature, diluted with THF (10 ml) and water (5 ml) then added LiOH·H₂O (150 mg, 3.51 mmol, 2.0 eq) and stirred for 4 h at room temperature. Progress of reaction was monitored by ELSD / TLC (SM was consumed). pH 3.0 of resulting reaction mixture was achieved by using IN HCI. The resulting reaction mixture was extracted with ethyl acetate (2 X 100 ml). Combined organic layer were washed with brine solution twice. The resulting organic layer was dried over sodium sulphate concentrated under reduced pressure to obtain crude which was purified by column chromatography by using 0-100% ethyl acetate in n-hexane to remove the nonpolar impurities, then used 10% MeOH in DCM gradient to obtain desired product [3] (2.55 g, 65.4%, Yield) as clear liquid and slowly transformed into white crystalline solid.Results:

[0350] ESI-MS analysis: Calculated C29H59NO4 [M+H] = 486.79, Observed = 486.8Intermediate [41

[0351] As depicted in Scheme 1: To a stirred solution of 4-(bis(2-((tert-butyldimethylsilyl)oxy) tridecyl)amino)-2-methylbutanoic acid (Intermediate [3], 1.5g, 3.09 mmol) in DCM (20 mL) was added the imidazole (1.26g, 18.53 mmol) and TBS-CI (2.79g, 18.53 mmol) at room temperature. The resulting reaction mixture was stirred at 35-40 °C for overnight (16-18h). Progress of reaction was monitored by ELSD / TLC (SM consumed). TLC showed completion of the starting material (TLC system: 40% ethyl acetate / pet-ether: 6c R / : 0.2, 6 R / : 0.6, Ninhydrin stain). The reaction mixture was cooled to room temperature and diluted with water (100 mL). The resulting reaction mixture was extracted with DCM (2 X lOOmL). Combined organic layer were washed with brine solution twice. The resulting organic layer was dried over sodium sulphate concentrated under reduced pressure to obtain crude which was purified by columnchromatography by using 0-100% ethyl acetate in n-hexane to obtain desired product [4] (2.8 g, 89%, Yield) as a colorless liquid.Results:

[0352] ESI-MS analysis: Calculated C41H87NO4Si2, [M+H] = 715.32, Observed = 715.4Intermediate [61

[0353] As depicted in Scheme 1: To a stirred solution of 4-(bis(2-((tert- butyldimethylsilyl)oxy)tridecyl)amino)-2-methylbutanoic acid (Intermediate 4, 1.5g, 2.06 mmol) in acetonitrile (20 mL) was added the potassium carbonate (570 mg, 4.12 mmol) and tert-butyl 4-(2-bromoethyl)piperazine-l-carboxylate [5] (724. 61 mg, 2.47 mmol). The resulting reaction mixture was stirred at 80°C for overnight. Progress of reaction was monitored by ELSD / TLC (SM consumed). After completion of the reaction, the reaction mixture was cooled to room temperature and filtered. The filtrate was concentrated under vacuum to give crude product. The crude was purified by flash chromatography (0 to 2.5 % MeOH in DCM) to give [6] (1.2g g, 62%) as a colorless liquid.Results:

[0354] ESI-MS analysis: Calculated C52H107N3O6Si2, [M+H] = 927.61, Observed = 827.7Intermediate [71

[0355] As depicted in Scheme 1: To a solution of [6] (1.2 g, 1.28 mmol) in DCM (20 mL) at 0 °C was added TFA (4.88 mL, 63.79 mmol). Then the reaction mixture was brought to room temperature and stirred for 14 hours. After completion of the reaction as monitored by MS. The reaction mixture was concentrated to obtain compound [7] with TFA salts (850 mg g, 80%). It was confirmed by MS analysis.Results:

[0356] NMR (400 MHz, CDCI3): δ4.32 - 4.05 (m, 2H), 3.59 (s, 2H), 3.08 (s, 3H), 2.67 (d, J= 18.0 Hz, 5H), 2.59 - 2.25 (m, 8H), 1.80 (s, 3H), 1.26 (s, 35H), 1.19 - 1.07 (m, 3H), 0.88 (d, J = 7.8 Hz, 24H), 0.06 (s, 12H).

[0357] ESI-MS analysis: Calculated C47H99N3O4Si2, [M+H] = 826.72, Observed = 826.7Intermediate [101

[0358] As depicted in Scheme 1: To stirred solution of 2-(piperazin-l-yl)ethyl 4-(bis(2-((tert- butyldimethylsilyl)oxy)dodecyl)amino)-2,2-dimethylbutanoate (intermediate 7) (800 mg, 0.95 mmol) in anhydrous dichloromethane (20 mL) added the ethylene sulfide [8] (74.38 mg, 1.24 mmol) at room temperature. The resulting reaction mixture was stirred for 24h and then added the pyridyl disulfide [9] (420 mg, 1.9 mmol) to the reaction mixture. Continued the stirring at room temperature for another 24 h. Monitored the reaction mixture using MS and after completion of the reaction concentrated to dryness under reduced pressure at below 45 °C. The obtained crude compound was purified by column chromatography using 100-200 mesh silica gel and eluent was 50% ethyl acetate / pet-ether to 100% ethyl acetate gives the desired compound

[0010] as yellow color oil (650 mg, 67%).Results:

[0359] ESI-MS analysis: Calculated C54H106N4O4S2Si2, [M+H] = 996.76, Observed = 996.7Intermediate [121

[0360] As depicted in Scheme 1: To a stirred solution of

[0010] (500 mg, 0.502 mmol) in chloroform (10 mL) were added triethylamine (280 pL, 2.01 mmol) and

[0011] (272.69 mg, 0.652 mmol) and stirred at room temperature for 2 hours. After completion of the reaction as monitored by MS. The reaction mixture was concentrated, and the crude compound was purified (eluent: 2-10% MeOH in DCM) to obtain pure compound

[0012] as a colorless oil (330 mg, 50%). It was confirmed by MS analysis.Results:

[0361] ESI-MS analysis: Calculated C73H152N4O6S2Si2, [M+H] = 1303.33, Observed = 1303.3Compound 1OH OHrNHOOH

[0362] As depicted in Scheme 1: To a solution of

[0012] (250 mg, 0.192 mmol) in THF (4 mL) was slowly added HF. Py (70% HF) (1.5 mL) at 0 °C. Then reaction mixture was brought to room temperature and stirred for 16 hours. After completion of the reaction as monitored by MS. The reaction mixture was concentrated, and the obtained residue was dissolved in ethyl acetate, washed with NaHCO₃ solution, water and brine. The organic layer was dried over anhydrous Na2SO4and concentrated. The crude product was purified to obtain Compound 1 (206 mg, 48%). It was confirmed by1H NMR and MS analysis.Results:

[0363] NMR (400 MHz, CDCI3) 64.24 (s, Hz, 3H), 3.81 (s, 4H), 3.22 - 2.26 (m, 34H), 1.53 - 1.06 (m, 67H), 0.81 (t, J = 6.4 Hz, 12H).

[0364] ESI-MS analysis: Calculated for C61H124N4O6S2, [M+H] = 1075.79; Observed = 11075.8Example IB. Synthesis of Compound 2

[0365] For example, Compound 2 may be prepared according to Scheme 2 (as depicted in Fig. 2).Example 1C. Synthesis of Compound 6

[0366] For example, Compound 6 may be prepared according to Scheme 3 (as depicted in Fig. 3).Intermediate (261

[0367] As depicted in Scheme 3: To a stirred solution of tert-butyl (2-aminoethyl) carbamate (1.0 g, 6.24 mmol, 1.0 eq) in methanol (20 ml), 1,2-epoxydecane (3.45 g, 18.72 mmol, 3.0 eq) was added followed by addition of DIPEA (2.02g, 15.60 mmol, 2.5 eq). The resulting reaction mixture was heated to 65°C for 16 h. Progress of reaction was monitored by ELSD / TLC (SM consumed). The resulting reaction mixture was cool down to room temperature and evaporated the volatiles. The resulting crude was diluted with water 100 mL and extracted with ethyl acetate (2 X 100 ml). Combined organic layer were washed with brine solution twice. The resulting organic layer was dried over sodium sulphate concentrated under reduced pressure to obtain crude which was purified by column chromatography by using 0-100% ethyl acetate in n-hexane to remove the nonpolar impurities, then used 10% MeOH in DCM gradient to obtain desired product

[0026] (2.56 g, 78%, Yield) as clear liquid.Results:

[0368] ESI-MS analysis: Calculated C31H64N2O4, [M+H] = 529.86, Observed = 529.8Intermediate [271

[0369] As depicted in Scheme 3: To a stirred solution of tert-butyl (2-(bis(2- hydroxydodecyl)amino)ethyl)carbamate (Intermediate

[0026] , 1.0g, 5.67 mmol) in DCM (20 mL) was added the imidazole (1.93g, 28.36 mmol) and TBS-CI (4.27g, 28.36 mmol) at room temperature. The resulting reaction mixture was stirred at 35-40 °C for overnight (16-18h). Progress of reaction was monitored by ELSD / TLC (SM consumed). TLC showed completion of the starting material (TLC system: 40% ethyl acetate / pet-ether: 6c R / : 0.2, 6 R / : 0.6, Ninhydrin stain). The reaction mixture was cooled to room temperature and diluted with water (100 mL). The resulting reaction mixture was extracted with DCM (2 X lOOmL). Combined organic layer were washed with brine solution twice. The resulting organic layer was dried over sodium sulphate concentrated under reduced pressure to obtain crude which was purified by column chromatography by using 0-100% ethyl acetate in n-hexane to obtain desired product

[0027] (3.2 g, 75%, Yield) as a colorless liquid.Results:

[0370] ESI-MS analysis: Calculated C43H92N2O4Si2, [M+H] = 758.39, Observed = 758.4Intermediate [281

[0371] As depicted in Scheme 3: To a solution of

[0027] (3.2 g, 4.23mmol) in DCM (25 mL) at 0 °C was added TFA (16.6 mL, 211.25 mmol). Then the reaction mixture was brought to room temperature and stirred for 14 hours. After completion of the reaction as monitored by MS. The reaction mixture was concentrated to obtain crude reaction mixture. The resulting mixture was diluted with saturated sodium bicarbonate solution (50 mL) and water (50 mL). The resulting reaction mixture was extracted with DCM (2 X 100mL). Combined organic layer were washed with brine solution twice. The resulting organic layer was dried over sodium sulphate concentrated under reduced pressure to obtain crude product

[0028] (2.2 g, 80%, Yield) as a color less liquid which is used for next steps without further purification. It was confirmed by MS analysis.Results:

[0372] ESI-MS analysis: Calculated C38H84N2O2Si2, [M+H] = 658.27, Observed = 658.2Intermediate [291

[0373] As depicted in Scheme 3: To a solution of 2-(4-(2-(pyridin-2-yldisulfaneyl)ethyl)piperazin-l- yl)ethan-l-ol (1.5 g, 5.01 mmol) and triethylamine (2.09 mL, 15.03 mmol) in dichloromethane (20 mL) at 0° C. was added 4-nitrophenyl-chloroformate (1.51 g, 7.51 mmol) and N, N-dimethyl aminopyridine (30 mg). The ice bath was removed and stirring continued for 7 h. The reaction was treated with an additional portion of diisopropylethylamine (1.0 mL), 4-nitrophenyl- chloroformate (1.5 g, 7.51 mmol), and N, N-dimethyl aminopyridine (30 mg) and stirred overnight. After completion of the reaction as monitored by MS, the reaction was washed with water, then IM potassium bisulfate, then saturated sodium bicarbonate, then brine, dried over magnesium sulfate, and concentrated to give 2.0 g (85%) as a light brown oil

[0029] , which was used immediately without purification.Results:

[0374] ESI-MS analysis: Calculated C2oH24N405S2, [M+H] = 465.56, Observed = 465.6Intermediate [301

[0375] As depicted in Scheme 3: To a solution of Nl, Nl-bis(2-((tert- butyldimethylsilyl)oxy)tridecyl)ethane-l,2-diamine

[0028] ( 0.5 g, 0.729 mmol) and triethylamine (0.254 mL, 1.8 mmol) in dichloromethane (10 mL) at 0° C. was added 4-nitrophenyl (2-(4-(2- (pyridin-2-yldisulfaneyl)ethyl)piperazin-l-yl)ethyl) carbonate

[0029] (0.677 g, 1.46 mmol) and N, N- dimethyl amino pyridine (10 mg). The ice bath was removed and stirring continued for 6h. After completion of the reaction as monitored by MS, the reaction was washed with water, then saturated sodium bicarbonate, then brine, dried over magnesium sulfate, and concentrated to give crude which was purified by column chromatography by using 0-100% ethyl acetate in n- hexane to remove the nonpolar impurities, then used 10% MeOH in DCM gradient to obtain desired product

[0030] (380 mg, 51%, Yield) as light yellow color liquid.Results:

[0376] ESI-MS analysis: Calculated C52H103N5O4S2Si2, [M+H] = 983.72, Observed = 983.7Intermediate [311

[0377] As depicted in Scheme 3: To a stirred solution of

[0030] (350 mg, 0.346 mmol) in chloroform (10 mL) were added triethylamine (157 pL, 1.13 mmol) and

[0011] (204.16 mg, 0.488 mmol) and stirred at room temperature for 2 hours. After completion of the reaction as monitored by MS.The reaction mixture was concentrated, and the crude compound was purified (eluent: 2-10% MeOH in DCM) to obtain pure compound

[0031] as a colorless oil (320 mg, 64%). It was confirmed by MS analysis.Results:

[0378] ESI-MS analysis: Calculated C71H149N5O6S2Si2, [M+H] = 1290.29, Observed = 1290.3Compound 6OH

[0379] As depicted in Scheme 3: To a solution of

[0031] (300 mg, 0.232 mmol) in THF (6 mL) was slowly added HF. Py (70% HF) (1.0 mL) at 0 °C. Then reaction mixture was brought to room temperature and stirred for 16 hours. After completion of the reaction as monitored by MS. The reaction mixture was concentrated, and the obtained residue was dissolved in ethyl acetate, washed with NaHCOa solution, water and brine. The organic layer was dried over anhydrous Na2SO4and concentrated. The crude product was purified to obtain Compound 6 (120 mg, 48%). It was confirmed by 1H-NMR and ELSD- MS analysis.Results:

[0380] TH NMR (400 MHz, CDCI3) 64.15 (s, 2H), 3.63 (m, 6H), 3.18 (s, 3H), 2.85 - 2.24 (m, 29H), 1.20 (s, 64H), 0.81 (s, 12H).

[0381] ESI-MS analysis: Calculated C59H121N5O6S2, [M + H+] = 1060.77; Observed = 1060.7Example ID. Synthesis of Compound 10

[0382] For example, Compound 10 may be prepared according to Scheme 4 (as depicted in Fig. 4).Intermediate (341

[0383] As depicted in Scheme 4: To a solution of 3-(bis(2-((tert- butyldimethylsilyl)oxy)dodecyl)amino)propan-l-ol

[0033] ( 0.5 g, 0.743 mmol) and triethylamine(0.207 mL, 1.8 mmol) in dichloromethane (10 mL) at 0° C. was added 4-nitrophenyl (2-(4-(2- (pyridin-2-yldisulfaneyl)ethyl)piperazin-l-yl)ethyl) carbonate

[0029] (0.691g, 1.49 mmol) and N, N- dimethyl amino pyridine (10 mg). The ice bath was removed and stirring continued for 6h. After completion of the reaction as monitored by MS, the reaction was washed with water, then saturated sodium bicarbonate, then brine, dried over magnesium sulfate, and concentrated to give crude which was purified by column chromatography by using 0-100% ethyl acetate in n- hexane to remove the nonpolar impurities, then used 10% MeOH in DCM gradient to obtain desired product

[0034] (335 mg, 45%, Yield) as colorless liquid.Results:

[0384] ESI-MS analysis: Calculated C53H104N4O5S2Si2, [M+H] = 998.73, Observed = 998.7Intermediate

[0035]

[0385] As depicted in Scheme 4: To a stirred solution of

[0030] (300 mg, 0.300 mmol) in chloroform (6 mL) were added triethylamine (150 pL, 1.13 mmol) and

[0011] (188.4 mg, 0.451 mmol) and stirred at room temperature for 2 hours. After completion of the reaction as monitored by MS. The reaction mixture was concentrated, and the crude compound was purified (eluent: 2-10% MeOH in DCM) to obtain pure compound

[0035] as a colorless oil (245 mg, 62.4%). It was confirmed by MS analysis.Results:

[0386] ESI-MS analysis: Calculated C72H150N4O7S2Si2, [M + H+] = 1305.30 Observed = 1305.3Compound 10OHOH HOOH

[0387] As depicted in Scheme 4: To a solution of

[0035] (280 mg, 0.232 mmol) in THF (6 mL) was slowly added HF. Py (70% HF) (0.7 mL) at 0 °C. Then reaction mixture was brought to room temperature and stirred for 16 hours. After completion of the reaction as monitored by MS. The reaction mixture was concentrated, and the obtained residue was dissolved in ethyl acetate, washed with NaHCO₃ solution, water and brine. The organic layer was dried over anhydrousNa2SO4and concentrated. The crude product was purified to obtain Compound 10 (110 mg, 47%). It was confirmed by ELSD- MS analysis.Results:

[0388] ESI-MS analysis: Calculated C60H122N4O7S2, [M + H+] = 1075.78 Results: Observed = 1075.8Example IE. Synthesis of Compound 4

[0389] For example, Compound 4 may be prepared according to Scheme 5 (as depicted in Fig. 5).Intermediate (381

[0390] As depicted in Scheme 5: To a solution of

[0037] (2 g, 7.74 mmol) in DCM (30 mL) were added

[0033] (5.99 g, 9.29 mmol) in DCM (25 mL), EDC (2.97 g, 15.48 mmol), DMAP (0.189 g, 1.55 mmol), DIPEA (4.03 mL, 23.23 mmol) and stirred at room temperature for 14 hours. After completion of the reaction as monitored by MS. The reaction mixture was diluted with DCM (200 mL) washed with NaHCO₃ solution, water and brine. The organic layer was dried over anhydrous Na₂SO₄U, concentrated, and the crude compound was purified (eluent: 20%-60% EtOAc in hexanes) to obtain pure compound

[0038] as a color less oil (4.8 g, 69%). It was confirmed by MS analysis. Results:

[0391] ESI-MS analysis: Calculated C49H87N3O6Si2, [M+H] = 685.06, Observed = 685.1Compound 4OH OH

[0392] As depicted in Scheme 5: To a 20 ml polypropylene scintillation vial was added the

[0041] (0.600 g, 0.465 mmol) along with 8 mL of dry tetrahydrofuran. The vial was cooled to 0-5 °C and HF. Py (70% HF) (1.1 mL, 41.68 mmol) was added dropwise. After addition, the reaction vial was allowed to warm to room temperature and stirred for 18 hours. Afterwards, the reaction mixture was cooled back to 0 °C and neutralized with solid sodium bicarbonate, diluted with dichloromethane washed with NaHCO₃ solution, water and brine. The organic layer was driedover anhydrous Na₂SO₄ and concentrated. The crude product was purified to obtain Compound 4 (0.493 g, 50.6%). It was confirmed by1H NMR and MS analysis.Results:

[0393] NMR (400 MHz, CDCI3) 65.31 (d, J = 5.8 Hz, 1H), 4.15 (s, 2H), 3.62 (s, 4H), 3.06 - 2.28 (m, 34H), 1.27 (s, 72H), 0.88 (d, J = 7.7 Hz, 12H).

[0394] ESI-MS analysis: Calculated C60H122N4O6S2, [M + H+] = 1060.88 Results: Observed = 1060.8Example IF. Synthesis of Compound 13

[0395] For example, Compound 13 may be prepared according to Scheme 6 (as depicted in Fig. 6).Intermediate

[0044]

[0396] As depicted in Scheme 6: To a solution of, 4-(2-((tert-butoxycarbonyl)amino)ethyl)-l- chloropiperazin-l-ium (2g, 7.5537 mmol) in anhydrous dichloromethane (20 mL) were added thiirane (590.34 mg, 9.8198 mmol) at room temperature. Stirred the reaction mixture for overnight and added the l,2-di(pyridin-2-yl) disulfane (500 mg, 22.661 mmol) and continued the stirring for another 6-7h. After completion of the reaction as monitored by MS. The reaction mixture was diluted with DCM washed with NaHCO₃ solution, water and brine. The organic layer was dried over anhydrous Na2SC>4, concentrated, and the crude compound was purified (eluent: 30% Ethyl acetate in Hexanes) to obtain pure compound

[0044] as a color less oil (1.54 g, 51.1%).

[0397] ESI-MS analysis: Calculated for C18H30N4O2S2, [M+H] = 399.58; Observed = 399.2Intermediate

[0045]

[0398] As depicted in Scheme 6: To a solution of, 4-(2-((tert-butoxycarbonyl)amino)ethyl)-l- chloropiperazin-l-ium (1.5g, 3.8 mmol) in anhydrous dichloromethane (15 mL) was slowly added the Trifluoro acetic acid (840 mg, 75 mmol) at room temperature. Stirred the reaction mixture 2-3h. After completion of the reaction as monitored by MS. The reaction mixture was diluted with DCM washed with NaHCO₃ solution, water and brine. The organic layer was dried over anhydrous Na2SO4, concentrated, and the crude compound

[0045] (950 mg, 85%) was pushed to next step. A small sample was purified for 1H-NMR analysis (eluent: 10% MeOH in DCM) to obtained pure compound as a color less oil. It was confirmed by 1H-NMR and MS analysis.

[0399] TH NMR (400 MHz, CDCI3): 68.39 (s, 1H), 7.61 (td, J = 14.8, 6.8 Hz, 2H), 7.05 (d, J = 8.0 Hz, 1H), 3.42 (q, J = 6.5 Hz, 4H), 3.09 - 2.85 (m, 5H), 2.85 - 2.71 (m, 2H), 2.57 (d, J = 23.5 Hz, 9H).

[0400] ESI-MS analysis: Calculated for CI3H22N4S2, [M+H] = 299.47; Observed = 299.1Intermediate [471

[0401] As depicted in Scheme 6: To a solution of 4-(bis(2-((tert- butyldimethylsilyl)oxy)dodecyl)amino)butanoic acid

[0046] (1.0 g, 1.428 mmol) in DCM (10 mL) were added 2-[4-[2-(2-pyridyldisulfanyl)ethyl]piperazin-l-yl]ethanamine

[0045] (511 mg, 1.7136 mmol) in DCM (5 mL), EDC (821.24 mg, 4.284 mmol), DMAP (34.89 mg, 0.2856 mmol), DIPEA (1.2 mL, 7.14 mmol) and stirred at room temperature for 16 hours. After completion of the reaction as monitored by MS. The reaction mixture was diluted with DCM washed with NaHCO₃ solution, water and brine. The organic layer was dried over anhydrous Na2SO4, concentrated, and the crude compound was purified (eluent: 10% MeOH in DCM) to obtain pure compound

[0047] as a color less oil (1.1 g, 79%). It was confirmed by 1H-NMR and MS analysis.Results:

[0402] XH NMR (400 MHz, CDCI3): 68.41 (s, 1H), 7.75 - 7.54 (m, 2H), 7.04 (s, 1H), 3.46 (d, J = 101.0 Hz, 4H), 3.02 - 2.04 (m, 22H), 1.72 (s, 4H), 1.21 (d, J = 7.1 Hz, 34H), 0.83 (d, J = 7.6 Hz, 24H).

[0403] ESI-MS analysis: Calculated C53H105N5O3S2Si2, [M+H] = 981.75, Observed = 981.6Intermediate [481

[0404] As depicted in Scheme 6: To a stirred solution of

[0047] (0.5g, 0.50 mmol) in chloroform (10 mL) were added triethylamine (170 pL, 1.15 mmol) and

[0011] (0.265 g, 0.61 mmol) and stirred at room temperature for 2 hours. After completion of the reaction as monitored by MS. The reaction mixture was concentrated, and the crude compound was purified (eluent: 2-10% MeOH in DCM) to obtain pure compound

[0048] as a colorless oil (325 mg, 49%). It was confirmed by MS analysis.Results:

[0405] ESI-MS analysis: Calculated for C72H151N5O5S2Si2, [M+H] = 1288.32; Observed = 1289.9Compound 13OH

[0406] As depicted in Scheme 6: To a 20 ml polypropylene scintillation vial was added the

[0047] (0.325 g, 0.211 mmol, 1.0 eq) along with 5 mL of dry tetrahydrofuran. The vial was cooled to 0-5 °C and HF / pyridine (1.1 mL, 41.68 mmol) was added dropwise. After addition, the reaction vial was allowed to warm to room temperature and stirred for 18 hours. Afterwards, the reaction mixture was cooled back to 0 °C and neutralized with solid sodium bicarbonate, diluted with dichloromethane washed with NaHCO₃ solution, water and brine. The organic layer was dried over anhydrous Na2SO4and concentrated. The crude product was purified to obtain Compound 13 (0.190 g, 68%). It was confirmed byTH NMR and MS analysis.Results:

[0407] TH NMR (400 MHz, CDCI3): 66.41 (s, 1H), 3.62 (s, 4H), 3.35 (s, 2H), 2.86 - 2.16 (m, 34H), 1.74 (d, J = 38.5 Hz, 7H), 1.27 (s, 64H), 0.88 (q, J = 5.7 Hz, 12H).

[0408] ESI-MS analysis: Calculated for C60H123N5O5S2, [M+H] = 1059.79; Observed = 1059.8Example 1G. Synthesis of Compound 16

[0409] For example, Compound 16 may be prepared according to Scheme 7 (as depicted in Fig. 7).Intermediate (501

[0410] As depicted in Scheme 7: To a stirred solution of

[0044] (2.0g, 5.02 mmol) in chloroform (20 mL) were added triethylamine (160 pL, 1.15 mmol) and

[0011] (2.31 g, 5.52 mmol) and stirred at room temperature for 2 hours. After completion of the reaction as monitored by MS. The reaction mixture was concentrated, and the crude compound was purified (eluent: 2-10% MeOH in DCM) to obtain pure compound

[0050] as a colorless oil (2.1g, 59%). It was confirmed by MS analysis.Results:

[0411] ESI-MS analysis: Calculated for C37H76N4O4S2, [M+H] = 705.16; Observed = 705.2Compound 16OH

[0412] As depicted in Scheme 7: To a solution of

[0052] (0.5 g, 0.64 mmol) in DMF (10 mL) were added intermediate

[0051] (0.46 g, 0.70 mmol), HATU (0.38g, 0.769 mmol), DIPEA (0.246 mL, 1.6 mmol.) and stirred at room temperature for 16 hours. After completion of the reaction as monitored by MS. The reaction mixture was diluted with DCM washed with ice cold water (2x20 mL) and brine (20 mL). The organic layer was dried over anhydrous Na2SO4, concentrated, and the crude compound was purified (eluent: 10% MeOH in DCM) to obtain pure Compound 16 as a color less oil (0.445 g, 50%). It was confirmed by 1H-NMR and MS analysis.Results:

[0413] 1H NMR (400 MHz, CDCl3) δ 4.06 (t, J = 6.8 Hz, 4H), 3.71 - 3.55 (m, 2H), 3.34 (t, J = 5.9 Hz, 2H), 2.88 - 2.15 (m, 35H), 1.87 - 1.16 (m, 100H), 0.87 (d, J = 7.1 Hz, 18H).

[0414] ESI-MS analysis: Calculated for C37H76N4O4S2, [M+H] = 1367.30; Observed = 1367.5Example 1H. Synthesis of Compound 17

[0415] For example, Compound 17 may be prepared according to Scheme 8 (as depicted in Fig. 8).Compound 17OH

[0416] As depicted in Scheme 8: To a solution of

[0054] (0.5 g, 0.66 mmol) in DMF (10 mL) were added intermediate

[0051] (0.46 g, 0.73 mmol), HATU (0.303g, 0.797 mmol), DIPEA (2.29 mL, 1.6 mmol.) and stirred at room temperature for 16 hours. After completion of the reaction as monitored by MS. The reaction mixture was diluted with DCM washed with ice cold water (2x20 mL) and brine (20 mL). The organic layer was dried over anhydrous Na2SO4, concentrated, and the crude compound was purified (eluent: 10% MeOH in DCM) to obtain pure Compound 17 as a color less oil (0.345 g, 38%). It was confirmed by 1H-NMR and MS analysis.Results:

[0417] NMR (400 MHz, CDCI3) 66.35 (s, 1H), 4.85 (q, J = 6.0 Hz, 1H), 4.05 (t, J = 6.5 Hz, 2H), 3.71 - 3.26 (m, 5H), 2.88 - 2.11 (m, 36H), 1.89 - 1.18 (m, 97H), 0.88 (t, J = 6.3 Hz, 15H).

[0418] ESI-MS analysis: Calculated for C37H76N4O4S2, [M+H] = 1339.35; Observed = 1339.4Example 1J. Synthesis of Compound 18

[0419] For example, Compound 18 may be prepared according to Scheme 9 (as depicted in Fig. 9).Compound 18OH

[0420] As depicted in Scheme 9: To a 20 ml polypropylene scintillation vial was added the

[0057] (0.30 g, 0.184 mmol, 1.0 eq) along with 5 mL of dry tetrahydrofuran. The vial was cooled to 0-5 °C and HF / pyridine (1.0 mL, 41.68 mmol) was added dropwise. After addition, the reaction vial was allowed to warm to room temperature and stirred for 18 hours. Afterwards, the reaction mixture was cooled back to 0 °C and neutralized with solid sodium bicarbonate, diluted with dichloromethane washed with NaHCO₃ solution, water and brine. The organic layer was dried over anhydrous Na2SO4and concentrated. The crude product was purified to obtain Compound 18 (0.110 g, 42.6%). It was confirmed by MS analysis.Results:

[0421] ESI-MS analysis: Calculated for CsoHisgNsOgSz, [M+H] = 1399.15; Observed = 1399.1Example IK. Synthesis of Compound 48

[0422] For example, Compound 48 may be prepared according to Scheme 10 (as depicted in Fig.10).Intermediate [3A1

[0423] As depicted in Scheme 10: To a stirred solution of piperazine (5 g, 2 eq., 58 mmol) in chloroform (50 ml) and methanol (50 mL), was added tert-butyl acrylate (3.72 g, 29 mmol). The reaction mass was allowed to stir at 45° for 16 h. The progress of reaction was monitored by ELSD / TLC (SM was consumed). The reaction mixture was concentrated to remove excess of methanol. Residue was diluted with water (50 mL) and extracted with DCM (2x 50 mL). The separated organic layer was dried over Na2SO4and concentrated under reduced pressure. The resulting crude was purified over silica using 5-15% MeOH in DCM to obtain [3A] (5 g, 80.39%), as a pale yellow liquid.Results:

[0424] ELSD analysis: Purity 97.85 %, Calculated Formula: C11H22N2O2 = 214.17, Observed = 215.00 (m / z, M+H+).Intermediate [5A]

[0425] As depicted in Scheme 10: A stirred solution of [3A] (5 g, 23.3 mmol) and thirane (1.54 g, 1.1 eq., 25.7 mmol) in toluene (50 mL) was heated to 80°C for 16h. The progress of the reaction was monitored by ELSD / TLC (SM was consumed). The reaction mixture was quenched with water (50 ml) and extracted with EtOAc (2x 25 mL). The separated organic layer was dried over Na2SO4and concentrated under reduce pressure. The resulting crude was purified over silica using 5% ethyl acetate in n-hexane to obtain [5A] (2.5 g, 39.05%).Results:

[0426] 1H-NMR (400MHz, CDCl3): δ 2.84 (t, J = 8.36 Hz, 2H), 2.69 (t, J = 7.52 Hz, 4H), 2.55 (s, 8H), 2.44 (t, J = 7.04 Hz, 2H), 1.45 (s, 9H).Intermediate [8A1

[0427] As depicted in Scheme 10: A stirred solution of triphenylmethanethiol (40 g, 145 mmol) in dimethylformamide (280 mL) was cooled to 0°C and sodium hydride 60% w / w (6.95 g, 1.2 eq., 174 mmol) added portion wise under a nitrogen atmosphere. The reaction was stirred for 15 min, then 2-(4-bromobutyl)-2,3-dihydro-lH-isoindole-l, 3-dione (40.8 g, 145 mmol) was added under an inert atmosphere at the same temperature. The reaction mass was allowed to stir at RT for 16 h. The progress of the reaction was monitored with TLC (SM was consumed completely). The reaction mixture was poured in ice cold water (2.0 Lit) and extracted with EtOAc (2 x 250 mL). The combined organic layer was washed with brine (3 x 250 mL) and dried over sodium sulphate. The organic layer was concentrated under reduced pressure. The crude compound was taken in ethanol and the precipitate filtered and dried to get [8A] (40 g, 50.9%) as an off white solid.Results:

[0428] 1H NMR (400 MHz, CDCl3): δ 8.07 (m, 2H), 7.51- 7.49 (m, 2H), 7.43-7.33 (m, 6H), 7.25 (t, J = 6.84 Hz, 6H), 7.18 (t, J = 7.12 Hz, 3H) 3.30 (m, 2H), 2.20 (t, J = 7.16 Hz, 2H), 1.56- 1.5 (m, 2H), 1.45-1.38 (m, 2H).Intermediate (9A1

[0429] As depicted in Scheme 10: To a stirred suspension of [8A] (40 g, 83.7 mmol) in ethanol (421 mL,) was added hydrazine hydrate (21 g, 5 eq., 419 mmol) at RT. The reaction was stirred at 90°C for 16h. The progress of the reaction was monitored by TLC (SM was consumed). The solid suspension was filtered through a sintered funnel and the filtrate was concentrated. The residuewas taken in DCM (500 mL), and a white precipitate came out which was again filtered off through a sintered funnel. The filtrate was concentrated under vacuum to get [9A] (27 g, 92.7%) as a light green viscous oil.Results:

[0430] NMR (400 MHz, CDCl3): δ 7.47-7.41 (m, 6H), 7.37-7.23 (m, 6H), 7.20-7.12 (m, 3H), 2.54 (t, J = 6.36 Hz, 2H), 2.16-2.10 (m, 2H), 1.43-1.22 (m, 4H).Intermediate (11A1x -, OHLkxx, SCPh3 / - - ■. ■ ' yJOH

[0431] As depicted in Scheme 10: To a stirred solution of [9A] (20 g, 57.5 mmol) in IPA (150 mL), was added 2-octyloxirane (22.5 g, 2.5 eq., 144 mmol) at RT, then the reaction mixture was refluxed at 90°C for 24 h. The progress of the reaction was monitored by TLC / ELSD. The reaction mixture was concentrated under reduced pressure and purified over a silica gel column using 2- 5% methanol in DCM to afford [11A] (13 g, 34.22%) as a colourless sticky liquid.Results:

[0432] ELSD analysis: Purity 99.23 %, Calculated C43H65NO2S = 659.47, Observed = 660.30 (m / z, M+H+).Intermediate (12A1„OTBSk xx, SCPh3N' YOTBS

[0433] As depicted in Scheme 10: To a stirred solution of [11A] (6 g, 1.1 eq., 9.09 mmol) in dichloromethane (60 mL), were added tert-butyl(chloro)bis(methyl)silane (10 g, 8 eq., 66.6 mmol) and imidazole (9.06 g, 16 eq., 133 mmol) at 0°C under nitrogen atmosphere. The reaction mass was allowed to stir at room temperature for 16h. The progress of reaction was monitoredby ELSD / TLC (SM was consumed). The reaction mixture was filtered through sintered funnel and water (100 mL) was added to the filtrate. The organic layer was extracted, dried over Na2SO4and concentrated under reduced pressure. The resulting crude was purified over silica using (1-5)% ethyl acetate in n-hexane to obtain [12A] (6 g, 81.15%) as a colourless liquid.Results:

[0434] ELSD analysis: Purity 79.23 %, Calculated C55H93NO2SSi2= 887.65, Observed = 889.10 (m / z, M+H+).Intermediate (13A1OTBS

[0435] As depicted in Scheme 10: To a stirred solution of [12A] (6 g, 6.75 mmol) in dichloromethane (60 mL) were added trifluoroacetic acid (12.9 mL, 25 eq., 169 mmol) and triethylsilane (1.29 mL, 1.2 eq., 8.1 mmol) at 0°C and the reaction mixture was allowed to stir for 10 min at RT. The reaction progress was monitored by TLC. SM was consumed completely. The reaction mass was quenched with aqueous sodium bicarbonate (100 mL) and extracted with DCM (2x 100 mL). The combined organic layer was dried over sodium sulphate and evaporated under reduced pressure to afford crude of [13A] (4.2 g, crude) as a colourless semisolid. The crude was forwarded to next step as such.Results:

[0436] ELSD analysis: Purity 92.21 %, Calculated C36H79NO2SSi2= 645.54, Observed = 646.70 (m / z, M+H+).Intermediate (15A1OTBS

[0437] As depicted in Scheme 10: To a stirred solution of [13A] (3.55 g, 5.49 mmol) in dichloromethane (40 mL). were added triethylamine (3.83 mL, 5 eq., 27.5 mmol) and 2-(2- pyridyldithio)pyridine (1.82 g, 1.5 eq., 8.24 mmol) at 0°C under a nitrogen atmosphere, then allowed to stir at RT for 16 h. The progress of the reaction was monitored by TLC / ELSD. The reaction mass was evaporated under reduced pressure and purified with silica gel flash column chromatography using 7-8 % MeOH in DCM to afford [15A] (3.5 g, 84.35%) as a colourless liquid.Results:

[0438] ELSD analysis: Purity 98.68 %, Calculated C4iH82N2O2S2Si2= 754.54, Observed = 755.95 (m / z, M+H+).Intermediate [16A]O TH SO,0 Y 'iN"OTBS

[0439] As depicted in Scheme 10: A stirred solution of [5A] (1 g, 3.64 mmol) and [15A] (2.75 g, 3.64 mmol) in dichloromethane (20 mL) was allowed to stir for 16 h. The reaction mass was allowed to stir at room temperature for 16 h. Progress of the reaction was monitored by ELSD / TLC (SM was consumed). Water (50 ml) was added to the reaction mixture and extracted with DCM (2x 30 mL). The separated organic layer was dried over Na2SO4and concentrated under reduced pressure. The resulting crude was purified over silica using 5% ethyl acetate in n-hexane to obtain [16A] (2.5 g, 74.68%) as a colourless liquid.Results:

[0440] ELSD analysis: Purity 97, 1 %, Calculated Formula: C4gHio3N304S2Si2= 917.69 Observed = 918.80 (m / z, M+H+).Intermediate [17A]TBSO' N'OTBS

[0441] As depicted in Scheme 10: To a stirred solution of [16A] (1 g, 1.09 mmol) in dichloromethane (20 mL), was added TFA (248 mg, 2 eq., 2.18 mmol) at 0°C. The reaction mass was allowed to stir at room temperature for 16h. Progress of the reaction was monitored by ELSD / TLC (SM was consumed). The reaction mass was concentrated under reduced pressure. The residue was diluted with DCM (2x 20 mL) and washed with aq. NaHCOg (2x 25 mL). The separated organic layer was dried over Na2SO4and concentrated under reduced pressure to afford [17A] (0.9 g, 95.85%) as a yellow oil.Results:

[0442] ELSD analysis: Purity 92.43 %, Calculated Formula: C45Hg5N3O4S2Si2 = 861.63, Observed = 862.75 (m / z, M+H+).Intermediate [20A]OH

[0443] As depicted in Scheme 10: A solution of 2-decyloxirane (13.2 g, 2.5 eq., 71.7 mmol) and tertbutyl (3-aminopropyl)carbamate (5 g, 28.7 mmol) in isopropanol (50 mL) was allowed to stir at 90°C for 16 h. Progress of the reaction was monitored by ELSD / TLC (SM was consumed). The reaction mass was concentrated under reduced pressure to obtain crude product [20A]. The resulting crude was forwarded to next step without further purification.Results:

[0444] ELSD analysis: Purity 99.82 %, Calculated Formula: C32H66N2O4 = 542.50, Observed = 543.45 (m / z, M+H+).Intermediate [21A],-OTBSvN^x-x^NHBocOTBS

[0445] As depicted in Scheme 10: To a stirred solution of [20A] (15.6 g, 28.7 mmol) in dichloromethane (150 mL), were added tert-butyl(chloro)bis(methyl)silane (34.6 g, 8 eq., 230 mmol) and imidazole (31.3 g, 16 eq 459 mmol). The reaction mass was allowed to stir at room temperature for 16 h. Progress of the reaction was monitored by ELSD / TLC (SM was consumed). The reaction mixture was quenched with water (500 mL) and extracted with DCM (3x 100 mL). The separated organic layer was dried over Na₂SO₄U and concentrated under reduced pressure. The resulting crude was purified over silica using 5% ethyl acetate in n-hexane to obtain [21A] (15.6 g, 68%) as a colourless liquid.Results:

[0446] ELSD analysis: Purity 95.06 %, Calculated Formula: C44H94N2O4Si2 = 770.68, Observed =771.85 (m / z, M+H+).Intermediate [22A],„OTBS"'NZ‘Z'X^'''NH2OTBS

[0447] As depicted in Scheme 10: To a stirred solution of [21A] (22.1 g, 28.7 mmol) in dichloromethane (150 mL), was added TFA (32.7 g, 10 eq., 287 mmol) at 0°C. The reaction mass was allowed to stir at room temperature for 16 h. Progress of the reaction was monitored by ELSD / TLC (SM was consumed). The reaction mixture was diluted with DCM (200 mL) and washed with a saturated solution of NaHCO₃ (2x 150 mL) followed by fresh water (2x 150 mL). The organic layer was dried over sodium sulphate and concentrated under reduced pressure. The resulting crude [22A] (20.0 g) was forwarded to next step without further purification.Results:

[0448] ELSD analysis: Purity 61.39 %, Calculated C39H86N2O2Si2= 670.62, Observed = 671.20 (m / z, M+H+).Intermediate (23A1OTB S

[0449] As depicted in Scheme 10: To a stirred solution of [17A] (900 mg, 1.04 mmol) in dichloromethane (25 mL) were added 2-methyl-2, 6, 8-triaza-6,7-decadiene— hydrogen chloride (1 / 1) (313 mg, 1.5 eq., 1.63 mmol) and N, N-dimethyl-4-pyridylamine (133 mg, 1.09 mmol) at room temperature. After 15 min, [22A] (731 mg, 1.09 mmol) was added to the resulting reaction mixture and allowed to stir at room temperature for 16 h. Progress of the reaction was monitored by ELSD / TLC (SM was consumed). The reaction mixture was quenched with water (500 mL) and extracted with DCM (3x 100 mL). The separated organic layer was dried over Na2SO4and concentrated under reduced pressure. The resulting crude was purified over silica using 5% ethyl acetate in n-hexane to obtain [23A] (1.2 g, 76%).

[0450] Note: One more batch was performed by the same process to get 2.0 g of [23A],Results:

[0451] ELSD analysis: Purity 97.55 %, Calculated Formula: C84Hi79N5OsS2Si4= 1514.24, Observed = 1515.90 (m / z, M+H+).Compound 48OH

[0452] As depicted in Scheme 10: To a stirred solution of [23A] (2 g, 1.32 mmol) in tetrahydrofuran (45 mL), was added hydrogen fluoride— pyridine (1 / 1) (1.31 g, 10 eq., 13.2 mmol) at 0°C. The reaction mass was allowed to stir at room temperature for 16 h. Progress of the reaction wasmonitored by ELSD / TLC (SM was consumed). The reaction mixture was diluted with diethyl ether (50 mL) and washed with water (2x 25 mL). The organic layer was washed with a saturated solution of NaHCO₃ x 25 mL), dried over Na₂SO₄ and concentrated under reduced pressure. The resulting crude was purified over silica using 5% MeOH in DCM to obtain Compound 48 (0.9 g, 64.43%).Results:

[0453] NMR (400 MHz, CDCl3): δ 8.24-8.20 (m, 1H), 3.69-3.36 (m, 4H), 3.51-3.43 (m, 1H), 3.36- 3.27 (m, 1H), 3.25-3.15 (m, 1H), 2.88-2.79 (m, 2H), 2.73-2.26 (m, 30H), 1.78-1.53 (m, 6H), 1.47- 1.22 (m, 66H), 0.87 (t, J= 6.32 Hz, 12H).

[0454] ELSD analysis: Purity 99.86 %, Calculated Formula: C60H123N5O5S2 = 1057.90, Observed = 1058.75 (m / z, M+H+).Example 1L. Synthesis of Compound 49

[0455] For example, Compound 49 may be prepared according to Scheme 11 (as depicted in Fig.11).Intermediate (10B1

[0456] As depicted in Scheme 11: The synthesis of [10B] is discussed in Example IK as Intermediate [ISA].Intermediate (13B1

[0457] As depicted in Scheme 11: To a stirred solution of 1,3-isoindolinedione (4 g, 27.2 mmol) in dimethylformamide (0.1 L), were added dipotassium carbonate (11.3 g, 3 eq., 81.6 mmol) and 1,7-dibromoheptane (21 g, 3 eq., 81.6 mmol) at room temperature, then allowed to stir at room temperature for 16h. Progress of the reaction was monitored by ELSD / TLC (SM was consumed). The reaction mixture was quenched with ice cold water (100 mL) and extracted with ethyl acetate (2x 50 mL). The collected organic layer was washed with (3x 50 mL) ice cold water. The separated organic layer was dried over Na2SO4and concentrated under reduced pressure. Theresulting crude was purified over silica gel using 5-20% ethyl acetate in heptane [13B] (6.2 g, 70.34%) as a colourless liquid.Results:

[0458] ELSD analysis: Purity 99.72 %, Calculated Formula: C₁₅H₁₈BrNO₂ = 323.05, Observed = 323.80 (m / z, M+H+).Intermediate (15B1

[0459] As depicted in Scheme 11: To a stirred solution of tert-butyl 1-piperazinecarboxylate (1 g, 5.37 mmol) in dimethylformamide (10 mL),. were added dipotassium carbonate (7.42 g, 10 eq., 53.7 mmol) and [13B] (2.26 g, 1.3 eq., 6.98 mmol) at RT, then the reaction mass was allowed to stir at room temperature for 16h. Progress of the reaction was monitored by ELSD / TLC. The reaction mixture was quenched with water (50 mL) and extracted with ethyl acetate (2x 20 mL). The organic layer was washed with water (3x 25 mL), dried over Na₂SO₄ and concentrated under reduced pressure. The resulting crude was purified over silica gel using (5-50%) ethyl acetate in n-hexane to obtain [15B] (1.4 g, 60.7%) as a pure product.Results:

[0460] ELSD analysis: Purity 99.83 %, Calculated Formula: C24H35N3O4 = 429.56, Observed = 430.10 (m / z, M+H+).Intermediate (16B1H2NxNL. ^NBoc

[0461] As depicted in Scheme 11: To a stirred solution of [15B] (1.4 g, 3.26 mmol) in ethanol (14 mL), was added diazane-water (1 / 1) (1.02 mL, 5 eq., 16.3 mmol) at room temperature, then the reaction mixture was heated at 80°C for 16h. Progress of the reaction was monitored by TLC & ELSD. After completion of the reaction, the mixture was filtered & washed by DCM. The filtrate was concentrated & filtered again. The filtrate was concentrated under reduced pressure. The resulting crude was diluted with water (25 mL) and extracted with 10% methanol in DCM (2x 25mL). The collected organic layer was dried over sodium sulfate and concentrated under reduced pressure to afford [16B] (0.8 g, 81.97%) as a colourless liquid.Results:

[0462] ELSD analysis: Purity 97.71 %, Calculated Formula: C16H33N3O2 = 299.26, Observed = 299.95 (m / z, M+H+).Intermediate (18B1OH

[0463] As depicted in Scheme 11: To a stirred solution of [16B] (0.8 g, 2.67 mmol) in isopropanol (7 mL), was added 2-decyloxirane (1.23 g, 2.5 eq., 6.68 mmol) and allowed to stir at 90°C for 16h. Reaction progress was monitored by TLC. The reaction mixture was evaporated under reduced pressure. The resulting crude was purified over silica gel by using 1-5% methanol in DCM to afford [18B] (1.7 g, 95.25%) as a colourless liquid.Results:

[0464] ELSD analysis: Purity 99.97 %, Calculated Formula: C40H81N3O4 = 667.62, Observed = 669.10 (m / z, M+H+).Intermediate [19B]OTBDMS

[0465] As depicted in Scheme 11: To stirred solution of [18B] (1.6 g, 2.39 mmol) in DCM (30 mL), were added imidazole (2.61 g, 16 eq., 38.3 mmol) & tert-butyl(chloro)bis(methyl)silane (2.89 g, 8 eq., 19.2 mmol), then allowed to stir at RT for 16h. Reaction progress was monitored by TLC and ELSD data. The reaction mass was quenched with water (100 mL) and extracted with DCM (3 x 50 mL). The organic layer was dried over sodium sulfate and evaporated under reducedpressure. The resulting crude was purified by silica gel flash column chromatography using 10 % EtOAc in heptane to afford [19B] (1.7 g, 79.17%) as a colourless liquid.Results:

[0466] ELSD analysis: Purity 99.94 %, Calculated Formula: C₅₂H₁₀₉N₃O₄Si₂ = 895.80, Observed = 897.15.00 (m / z, M+H+).Intermediate [20B]OTBDMS

[0467] As depicted in Scheme 11: To a stirred solution of [19B] (0.8 g, 892 pmol) in dichloromethane (15 mL), was added TFA (1.37 mL, 20 eq., 17.8 mmol) at 0°C. The reaction mass was allowed to stir at room temperature for lh. Progress of the reaction was monitored by ELSD / TLC (SM was consumed). The reaction mixture was concentrated under reduced pressure with repeated addition of DCM (3x 5 mL) to [20B] (430 mg, 60.52%) as a yellow gummy mass.Results:

[0468] ELSD analysis: Purity 99.56 %, Calculated Formula: C₄₇H₁₀₁N₃O₂Si₂ = 795.74, Observed = 797.00 (m / z, M+H+).Intermediate [22B]BrxX'S.'SCPh3

[0469] As depicted in Scheme 11: To a stirred solution of triphenylmethanethiol (25 g, 90.4 mmol) in ethanol (0.2 L) and water (0.2 L), was added sodium hydroxide (7.24 g, 2 eq., 181 mmol). After addition, the reaction mass was left for 30 min at room temperature, then 1,2-dibromoethane (25.5 g, 1.5 eq., 136 mmol) was added dropwise. The reaction mixture was allowed to stir at room temperature for 16 h. Reaction progress was monitored by TLC (after SM consumed). The reaction mixture was evaporated to remove excess ethanol. The residue was diluted with DCM (100 ml) and washed with water (2x 50 ml). The organic layer was dried over anhydrous sodiumsulfate and distilled out under reduced pressure. The crude was crystalized in methanol to afford [22B] (30 g, 86.52%) as an off white solid.Results:

[0470] NMR (400 MHz, CDCl3): δ 7.50-7.43 (d, 7 = 7.52 Hz, 6H), 7.36-7.23 (m, 9H), 2.89 (t, J = 7.56 Hz, 2H), 2.74 (t, J = 8.52 Hz, 2H).Intermediate (23B1'xSCPh3

[0471] As depicted in Scheme 11: To a stirred solution of [20B] (1.2 g, 1.51 mmol) in dimethylformamide (12 mL) were added dipotassium carbonate (625 mg, 3 eq., 4.52 mmol) and [22B] (866 mg, 1.5 eq., 2.26 mmol) at 0°C. The reaction mass was allowed to stir at room temperature for 16 h. Progress of the reaction was monitored by ELSD / TLC (SM was consumed). The reaction mixture was quenched with ice cold water (50 ml) and extracted with ethyl acetate (3x 25 mL). The separated organic layer was dried over Na2SO4and concentrated under reduced pressure. The resulting crude was purified over silica using (3-15%) ethyl acetate in n-hexane to obtain [23B] (0.9 g, 54.36%) as a colourless viscous liquid.Results:

[0472] ELSD analysis: Purity 99.53%, Calculated Formula: C₆₅H₁₁₉N₃O₄SSi₂ = 1097.86, Observed = 1099.25 (m / z, M+H+).Intermediate [24B]O'i'BDMS

[0473] As depicted in Scheme 11: To a stirred solution of [23B] (0.8 g, 728 pmol) in dichloromethane (8 mL), were added triethylsilane (140 pL, 1.2 eq., 874 pmol) & trifluoroacetic acid (1.39 mL, 25 eq., 18.2 mmol) at 0°C and the reaction mixture was allowed to stir for 10 min.Reaction progress was monitored by TLC. SM was consumed completely. The reaction mass was evaporated under reduced pressure with repeated addition of DCM (3x 20 mL) to afford [24B] (620 mg, 99%) as a colourless semisolid. The crude was forwarded to next step without purification.Results:

[0474] ELSD analysis: Purity 96.75 %, Calculated Formula: C₄₉H₁₀₅N₃O₂SSi₂ = 855.75, Observed = 857.00 (m / z, M+H+).Intermediate (25B1QTHDM5

[0475] As depicted in Scheme 11: To a stirred solution of [24B] (620 mg, 724 pmol) in dichloromethane (11.2 mL), was added [10B] (547 mg, 724 pmol) at room temperature under a nitrogen atmosphere. The reaction mass was allowed to stir for 16 h. Progress of the reaction was monitored by TLC / ELSD. The reaction mass was evaporated under reduced pressure and purified over a silica gel flash column using 5-30 % ethyl acetate in n-heptane to afford [25B] (0.3g, 27.62%) as a yellow liquid.Results:

[0476] ELSD analysis: Purity 98.64 %, Calculated Formula: C85Hi82N4O4S2Si4= 1499.27, Observed = 1501.15 (m / z, M+H+).Compound 49OH

[0477] As depicted in Scheme 11: To a stirred solution of [25B] (0.3 g, 0.2 mmol) in tetrahydrofuran (10.0 mL), was added hydrogen fluoride— pyridine (1 / 1) (540 pL, 30 eq., 6 mmol) at roomtemperature, then allowed to stir for 16h. Progress of the reaction was monitored by TLC / ELSD. The reaction mixture was diluted with diethyl ether (20 mL) and washed with water (2x 25 mL). The organic layer was further washed with sat. NaHCO₃ solution (2x 15 mL) followed by water (15 mL), dried over anhydrous Na₂SO₄ and concentrated under reduced pressure. Residue was dissolved in pentane (5 mL) and washed with ACN (5 mL). The pentane layer was evaporated under reduced pressure. The resulting crude was purified over silica gel flash chromatography by using 4-7% methanol in DCM to afford Compound 49 (145 mg, 69.05%) as a light yellow liquid.Results:

[0478] NMR (400 MHz, CDCl3): δ 3.72-3.58 (m, 4H), 2.83-2.79 (m, 2H), 2.71-2.67 (m, 4H), 2.65- 2.50 (m, 12H), 2.48-2.34 (m, 10H), 1.78-1.10 (m, 82H), 0.87 (t, J= 6.64 Hz, 12H).

[0479] ELSD analysis: Purity 99.38 %, Calculated Formula: C61H126N4O4S2 = 1042.92, Observed = 1044.50 (m / z, M+H+).Example IM. Synthesis of Compound 50

[0480] For example, Compound 50 may be prepared according to Scheme 12 (as depicted in Fig.12).Intermediate (3C1O

[0481] As depicted in Scheme 12: A stirred solution of tert-butyl 1-piperazinecarboxylate (7 g, 37.6 mmol) and thirane (2.46 mL, 1.1 eq., 41.3 mmol) in toluene (70 mL) was allowed to stir at 90°C for 24h. Reaction progress was monitored by TLC. The reaction mixture was evaporated under reduced pressure, and the resulting crude was purified over silica gel by using (5-50%) ethyl acetate in heptane to afford [3C] (3.2 g, 35%) as a colourless liquid.Results:

[0482] 1H-NMR (400MHz, CDCl3): δ 5.52-5.35 (m, 4H). 2.83-2.81 (d J = 4.0 Hz, 2H), 2.69-2.68 (d, 2H), 2.52-2.42 (m, 4H), 0.87 (s, 9H).Intermediate [13C]

[0483] As depicted in Scheme 12: The synthesis of [13C] is discussed in Example IK as Intermediate [ISA].Intermediate (14C1l O" J. ^x TBSO.. xx / -x,- x x-- -Q N; XL N •. S J'S' N'OTBS

[0484] As depicted in Scheme 12: A solution of [3C] (1 g, 4.06 mmol) and [13C] (3.07 g, 4.06 mmol) in dichloromethane (30 mL) was allowed to stir at room temperature for 16h. Progress of the reaction was monitored by ELSD / TLC (SM was consumed). Water (50 ml) was added to the reaction mixture and extracted with DCM (2x 30 mL). The separated organic layer was dried over Na₂SO₄ and concentrated under reduced pressure. The resulting crude was purified over silica using (5-50%) ethyl acetate in n-hexane to obtain [14C] (2.2 g, 61%) as a colourless liquid.Results:

[0485] ELSD analysis: Purity 100 %, Calculated Formula: C₄₇H₉₉N₃O₄S₂Si₂ = 889.66, Observed =890.15 (m / z, M+H+).Intermediate [15C]TBSO..HNNs'"sNOTBS

[0486] As depicted in Scheme 12: To a stirred solution of [14C] (0.9 g, 1.01 mmol) in dichloromethane (18.6 mL), was added TFA (230 mg, 2 eq., 2.02 mmol) at 0°C. The reaction mass was allowed to stir at room temperature for 6 h. Progress of the reaction was monitored by ELSD / TLC (SM was consumed). The reaction mixture was concentrated under reduced pressure. The residue was diluted with DCM (25 mL) and washed with saturated sodium bicarbonate solution until pH 7-8. The separated organic layer was dried over Na2SO4and concentratedunder reduced pressure to obtain [15C] (790 mg, crude). The resulting crude was forwarded the the next step without further purification.Results:

[0487] ELSD analysis: Purity 99.69 %, Calculated Formula: C42HgiN3O2S2Si2 = 789.61, Observed = 791.0 (m / z, M+H+).Intermediate (17C1OTBS

[0488] As depicted in Scheme 12: To a stirred solution of [15C] (790 mg, 999 pmol) in dimethylformamide (8 mL), were added N-ethyldiisopropylamine (690 pL, 4 eq., 4 mmol) and 2- iodoethanol (258 mg, 1.5 eq., 1.5 mmol) at room temperature. The reaction mass was allowed to stir at room temperature for 16 h. Progress of the reaction was monitored by ELSD / TLC (SM was consumed). The reaction mixture was quenched with ice cold water (25 mL) and extracted with diethyl ether (2x 25 mL). The separated organic layer was dried over Na2SO4and concentrated under reduced pressure. The resulting crude was purified over silica using (5-50%) ethyl acetate in n-hexane to obtain [17C] (530 mg, 64%) as pure product.Results:

[0489] ELSD analysis: Purity 99.70 %, Calculated Formula: C₄₄H₉₅N₃O₃S₂Si₂ = 833.64, Observed = 834.75 (m / z, M+H+).Intermediate [22C]

[0490] As depicted in Scheme 12: The synthesis of [22C] is discussed in Example IK as Intermediate[22A],Intermediate [23C]OTBS.oms IM I'""XI\T A. J " N N" 'O' TBSO''0TBS

[0491] As depicted in Scheme 12: To a stirred solution of [17C] (440 mg, 474 pmol) in tetrahydrofuran (7 mL) were added triethylamine (132 pL, 2 eq., 948 pmol) & [22C] (318 mg, 474 pmol) at RT, then allowed to stir at RT for lh. Reaction progress was monitored by TLC. The reaction mixture was evaporated under reduced pressure. The residue was dissolved in ethyl acetate (15 mL) and washed with water (2x 10 mL). The combined organic layer was dried over sodium sulfate and evaporated under reduced pressure. The resulting crude was purified over silica gel by using 1-5% methanol in DCM to afford [23C] (430 mg, 59.24%) as a colourless liquid.Results:

[0492] ELSD analysis: Purity 99.77 %, Calculated C₈₄H₁₇₉N₅O₆S₂Si₄ = 1530.24, Observed = 1531.80 (m / z, M+H+).Compound 50OH

[0493] As depicted in Scheme 12: To a stirred solution of [23C] (390 mg, 255 pmol) in tetrahydrofuran (6 mL), was added hydrogen fluoride— pyridine (1 / 1) (688 pL, 30 eq., 7.64 mmol) dropwise at room temperature, then was allowed to stir for 16h. Progress of the reaction was monitored by TLC / ELSD. The reaction mixture was diluted with diethyl ether (25 mL) and washed with water (2x 20 mL). The organic layer was further washed with sat. NaHCOa solution (2x 10 mL), followed by water (2x 10 mL). The separated organic layer was dried over anhydrous Na2SO4and concentrated under reduced pressure. The residue was dissolved in pentane (10 mL) and washed with ACN (2x 10 mL). The pentane layer was evaporated under reduced pressure. The resulting crude was purified by using 4-7% methanol in DCM to afford Compound 50 (240 mg, 87.71%) as a light yellow liquid.Results:

[0494] TH NMR (400 MHz, CDCI3): 66.12-5.84 (m, 1H), 4.34-4.00 (m, 2H), 3.74-3.54 (m, 4H), 3.48- 3.14 (m, 3H), 2.84-2.78 (m, 2H), 2.73-2.48 (m, 18H), 2.48-2.37 (m, 6H), 2.32- 2.25 (m, 1H), 1.80- 1.48 (m, 8H), 1.47-1.20 (m, 66H), 0.87 (t, J = 6.56 Hz, 12H).

[0495] ELSD analysis: Purity 99.73%, Calculated Formula: C60H123N5O6S2 = 1073.89, Observed = 1075.30 (m / z, M+H+).Example IN. Synthesis of Compound 51

[0496] For example, Compound 51 may be prepared according to Scheme 13 (as depicted in Fig.13).Intermediate [20D]

[0497] As depicted in Scheme 13: The synthesis of [20D] is discussed in Example IK as Intermediate [ISA].Intermediate [3D]N^^NHBOC

[0498] As depicted in Scheme 13: A stirred solution of 2-decyloxirane (14.4 g, 2.5 eq., 78 mmol) and tert-butyl (2-aminoethyl)carbamate (5 g, 31.2 mmol) in isopropanol (50 mL) was heated to 90°C for 16 h. Progress of the reaction was monitored by ELSD / TLC (SM was consumed). The solvent was concentrated under reduced pressure to obtain crude of [3D] (9 g, 55%) as a yellowed liquid, which was forwarded to the next step without further purification.Results:

[0499] ELSD analysis: Purity 99.94 %, Calculated Formula: C31H64N2O4 = 528.49, Observed = 529.45 (m / z, M+H+).Intermediate [4D]XOTBDMS. NHBocNOTBDMS

[0500] As depicted in Scheme 13: To a stirred solution of [3D] (9 g, 17 mmol) in dichloromethane (109 mL), were added tert-butyl(chloro)bis(methyl)silane (20.5 g, 8 eq., 136 mmol) and imidazole (18.5 g, 16 eq., 272 mmol) at 0 °C under a nitrogen atmosphere, then the reaction mass was allowed to stir at room temperature for 16 h. Progress of the reaction was monitored by ELSD / TLC (SM was consumed). The reaction mixture was diluted with water (500 mL) and extracted with DCM (3x100 mL). The separated organic layer was dried over Na₂SO₄ and concentrated under reduced pressure. The resulting crude was purified over silica using 5% ethyl acetate in n-hexane to obtain [4D] (10.2g, 79%) as a clear liquid.Results:

[0501] ELSD analysis: Purity 99.91 %, Calculated Formula: C43H92N2O4Si2 = 756.66, Observed =757.95 (m / z, M+H+).Intermediate [5D]OTBDMS

[0502] As depicted in Scheme 13: To a stirred solution of [4D] (10 g, 13.2 mmol) in dichloromethane (84.6 mL), was added trifluoroacetic acid (15.1 g, 10 eq., 132 mmol) at 0°C. The reaction mass was allowed to stir at room temperature for 6 h. Progress of the reaction was monitored by ELSD / TLC (SM was consumed). The reaction mass was concentrated under reduced pressure. The resulting residue was dissolved in DCM (50 mL) and then washed with saturated solution of NaHCO₃ (3x 25 mL). The organic layer was separated and dried over sodium sulphate and concentrated under reduced pressure to afford [5D] (7 g, 81%).Results:

[0503] CAD analysis: Purity 91.12 %, Calculated Formula: C38H84N2O2Si2 = 656.61, Observed = 657.77 (m / z, M+H+).Intermediate (7D1 / X. „, OTBDMSTH H. x- x_,.... / 'xTJ O L „NBocOTBDMS

[0504] As depicted in Scheme 13: To a stirred solution of tert-butyl 4-(2-aminoethyl)-l- piperazinecarboxylate (0.8 g, 3.49 mmol) in tetrahydrofuran (8 mL, 98.3 mmol) was added bis(l- imidazolyl)methanone (1.13 g, 2 eq., 6.98 mmol) at RT and the reaction mixture was stirred for lh at RT. The reaction mixture was monitored by TLC and a new non polar UV active spot was formed. The reaction mixture was evaporated under reduced pressure. The residue was diluted with ethyl acetate (30 mL) and washed with water (2x 25 mL). The combined organic layer was dried over sodium sulfate and evaporated under reduced pressure to get crude of tert-butyl 4- [2-(l-imidazolylcarbonylamino)ethyl]-l-piperazinecarboxylate (1.1 g,) as a colourless liquid. To the solution of resulting crude intermediate (1.1 g, 3.4 mmol) in tetrahydrofuran (20 mL, 246 mmol), were added [5D] (2.24 g, 3.4 mmol) and triethylamine (1.42 mL, 3 eq., 10.2 mmol) at RT then allowed to stir at RT for 16 h. Progress of the reaction was monitored by TLC. The reaction mixture was diluted with ethyl acetate (50 mL) and then washed with water (2x 30 mL). The combined organic layer was dried over sodium sulfate and evaporated under reduced pressure. The resulting crude was purified by silica gel flash column chromatography using (2-5% methanol in DCM) to afford [7D] (1.3 g, 41.8%) as a colourless liquid.Results:

[0505] ELSD analysis: Purity 99.10 %, Calculated Formula: C50H105N5O5Si2 = 911.77, Observed =912.95 (m / z, M+H+).Intermediate [8D]OTBDMS

[0506] As depicted in Scheme 13: To a stirred solution of [7D] (1.3 g, 1.42 mmol) in dichloromethane (26.2 mL), was added TFA (2.18 mL, 20 eq., 28.5 mmol) at 0°C. The reaction mass was allowed to stir at room temperature for 16 h. Progress of the reaction was monitored by ELSD / TLC (SM was consumed). The reaction mixture was concentrated under reduced pressure with repeated addition of DCM (3x 5 mL) and dried under high vacuum to afford [8D] (1.1 g, crude), which was forwarded to the next step as is.Results:

[0507] ELSD analysis: Purity 99.61%, Calculated Formula: C45H97N5O3Si2 = 811.71, Observed = 812.90 (m / z, M+H+).Intermediate [IIP]SCPhj

[0508] As depicted in Scheme 13: To a stirred solution of triphenylmethanethiol (25 g, 90.4 mmol) in ethanol (0.2 L) and water (0.2 L), was added sodium hydroxide (7.24 g, 2 eq., 181 mmol). After addition, the reaction mass was left for 30 min at room temperature, then 1,2-dibromoethane (25.5 g, 1.5 eq., 136 mmol) was added dropwise. The reaction mixture was allowed to stir at room temperature for 16 h. Reaction progress was monitored by TLC (after SM consumed). The reaction mixture was evaporated to remove excess ethanol. The residue was diluted with DCM (100 ml) and washed with water (2x 50 ml). The organic layer was dried over anhydrous sodium sulfate and distilled out under reduced pressure. The crude was crystalized in methanol to afford [11D] (30 g, 86.52%) as an off white solid.Results:

[0509] TH NMR (400 MHz, CDCl3): δ 7.50-7.43 (d, 7 = 7.52 Hz, 6H), 7.36-7.23 (m, 9H), 2.89 (t, J = 7.56 Hz, 2H), 2.74 (t, J = 8.52 Hz, 2H).Intermediate (12D1,. OTBDMSI H HkXN„, NN J O N SCPh3OTBDMS

[0510] As depicted in Scheme 13: To a stirred solution of [8D] (1.1 g, 1.35 mmol) in dimethylformamide (10 mL), were added dipotassium carbonate (936 mg, 5 eq., 6.77 mmol) and [11D] (779 mg, 1.5 eq., 2.03 mmol) at 0°C, then allowed to stir at room temperature for 16 h. Progress of the reaction was monitored by ELSD / TLC. The reaction mixture was quenched with ice cold water (25 mL) and extracted with ethyl acetate (2x 20 mL). The separated organic layer was dried over Na₂SO₄U and concentrated under reduced pressure. The resulting crude was purified over silica using (3-7%) methanol in DCM to obtain [12D] (0.9 g, 60%) as pure product.Results:

[0511] ELSD analysis: Purity 55.75 %, Calculated Formula: C66H118N5O4SSi2 = 1113.83, Observed = 1115.30 (m / z, M+H+).Intermediate [13D]OTBDMS

[0512] As depicted in Scheme 13: To a stirred solution of [12D] (0.9 g, 807 pmol) in dichloromethane (9 mL, 141 mmol), were added triethylsilane (155 pL, 1.2 eq., 969 pmol) & trifluoroacetic acid (1.54 mL, 25 eq., 20.2 mmol) at 0°C and the reaction mixture was allowed to stir for 5 min. Reaction progress was monitored by TLC. SM was consumed completely. The reaction mass was directly evaporated under reduced pressure to get crude [13D] (0.7 g) as a colourless semisolid, which was forwarded to the next step as such.Results:

[0513] ELSD analysis: Purity 35.30%, Calculated Formula: C47H101N5O3SSi2 = 871.72, Observed = 872.10 (m / z, M+H+).Intermediate [21D]. OTBDMS TBDMSO,.OTBDMSOTBOMS

[0514] As depicted in Scheme 13: To a stirred solution of [13D] (0.7 g, 802 pmol) in dichloromethane (7 mL), was added [20D] (606 mg, 802 pmol) and allowed to stir at room temperature for 16 h. Progress of the reaction was monitored by TLC / ELSD. The reaction mass was evaporated under reduced pressure and the resulting crude was purified with silica gel flash column chromatography using 2-7% methanol in DCM to afford [21D] (0.3 g, 24.65%) as a yellow viscous gummy mass.Results:

[0515] ELSD analysis: Purity 99.79 %, Calculated C83H178N6O5S2Si4 = 1515.24, Observed = 1517.10 (m / z, M+H+).Compound 51.. OH H HXN., HO..6 '’N'

[0516] As depicted in Scheme 13: To a stirred solution of [21D] (0.3 g, 198 pmol) in tetrahydrofuran (6.98 mL), was added hydrogen fluoride— pyridine (1 / 1) (535 pL, 30 eq., 5.93 mmol) dropwise at room temperature. Then reaction mixture was allowed to stir at room temperature for 16h. The reaction mixture was diluted with diethyl ether (20 mL) and washed with water (2x 15 mL). The separated organic layer was further washed with sat. NaHCO₃ solution (2x 15 mL) followed by water (2x 15 mL). The organic layer was dried over anhydrous Na2SO4and concentrated under reduced pressure. The residue was dissolved in pentane (15 mL) and washed with ACN (2x 10 mL). The pentane layer was evaporated under reduced pressure. The resulting crude was purified by silica gel flash chromatography by using 4-7% methanol in DCM to afford Compound 51 (150 mg, 71.56%) as a light yellow liquid.Results:

[0517] TH NMR (400 MHz, CDCI3): 65.77-5.46 (m, 2H), 3.70-3.54 (m, 4H), 3.35-3.18 (m, 4H), 3.00- 2.26 (m, 28H), 1.79-1.65 (m, 2H), 1.64-1.55 (m, 2H), 1.50-1.18 (s, 68H), 0.86 (t, J = 7.0 Hz, 12H).

[0518] ELSD analysis: Purity 99.61 %, Calculated C59H122N6O5S2 = 1058.89, Observed = 1060.40 (m / z, M+H+).Example 10. Synthesis of Compound 43

[0519] For example, Compound 43 may be prepared according to Scheme 14 (as depicted in Fig.14).Intermediate 9E

[0520] As depicted in Scheme 14: The synthesis of [9E] is discussed in Example IK as Intermediate [ISA].Intermediate [HE]OH

[0521] As depicted in Scheme 14: To a stirred solution of 2-decyloxirane (15.1 g, 2.5 eq., 81.9 mmol) and 2-aminoethanol (2 g, 32.7 mmol) in isopropanol (25 mL) was added and heated to 90°C. The reaction mass was allowed to stir at 90 °C for 16 h. Progress of the reaction was monitored by ELSD / TLC (SM was consumed). The reaction mixture was concentrated under reduced pressure. The resulting crude [11E] (14.0 g, 99.5%) was forwarded to the next step without further purification.Results:

[0522] ELSD analysis: Purity 95.13 %, Calculated Formula: C25H53NO3 = 429.42, Observed = 430.30 (m / z, M+H+).Intermediate [12E]L, OTritNOH

[0523] As depicted in Scheme 14: To a stirred solution of [HE] (14.0 g, 32.7 mmol) in dichloromethane (0.1 L), were added chlorotriphenylmethane (18.3 g, 2 eq., 65.5 mmol) and pyridine (15.5 g, 6 eq., 196 mmol). The reaction mass was allowed to stir at RT for 16 h. Progress of the reaction was monitored by ELSD / TLC (SM was consumed). The reaction mixture was concentrated under reduced pressure. The resulting crude [12E] (21.0 g, 95.22%) was forwarded to next step without further purification.Results:

[0524] ELSD analysis: Purity 82.12 %, Calculated Formula: C45H69NO3 = 671.53, Observed = 672.20 (m / z, M+H+).Intermediate [13E],-OTritNOTBDMS

[0525] As depicted in Scheme 14: To a stirred solution of [12E] (22 g, 32.7 mmol) in dichloromethane (0.1 L), were added tert-butyl(chloro)bis(methyl)silane (39.5 g, 8 eq., 262 mmol) and imidazole (35.7 g, 16 eq., 524 mmol) at 0 °C under nitrogen atmosphere. The reaction mass was allowed to stir at RT for 16 h. Progress of the reaction was monitored by ELSD / TLC (SM was consumed). The reaction mixture was concentrated under reduced pressure. The resulting crude was purified over silica gel using 0-15% EtOAc in heptane to afford [13E] (19 g, 64.44%) as a yellow oil.Results:

[0526] ELSD analysis: Purity 78.53%, Calculated Formula: C57H97NO4Si2 = 899.70, Observed = 900.20 (m / z, M+H+).Intermediate [14E]

[0527] As depicted in Scheme 14: To a stirred solution of [13E] (19 g, 21.1 mmol) in dichloromethane (64.4 mL), were added triethylsilane (4.05 mL, 1.2 eq., 25.3 mmol) and trifluoroacetic acid (40.4 mL, 25 eq., 527 mmol) successively at 0°C under a nitrogen atmosphere. The reaction mass was stirred for 5 min at the same temperature. Progress of the reaction was monitored by TLC (SM was consumed). The reaction mass was concentrated under reduced pressure. The residue was dissolved in DCM (100 mL) and washed with saturated solution of NaHCO₃ (2x 50 mL). The organic layer was dried over Na₂SO₄ and concentrated under reduced pressure. The resulting crude was purified over silica gel column using 5% ethyl acetate in hexane to obtain [14E] (12 g, 86.41%) as a colourless liquid.Results:

[0528] ELSD analysis: Purity 99.55%, Calculated Formula: C35H73NO3Si = 657.59, Observed = 658.55 (m / z, M+H+).Intermediate (16E1H X O, N.N' Y NOTBDMS

[0529] As depicted in Scheme 14: To a stirred solution of [14E] (4 g, 6.08 mmol) in tetrahydrofuran (40 ml) was added bis(l-imidazolyl)methanone (1.48 g, 1.5 eq., 9.11 mmol) followed by triethylamine (1.69 mL, 2 eq., 12.2 mmol). The reaction mass was allowed to stir for lh. Progress of the reaction mass was monitored by ELSD. SM was consumed. The reaction mixture was concentrated under reduced pressure. The crude was dissolved in ethyl acetate (50 mL) and washed with water (2x 30 mL). The organic layer was dried over sodium sulphate, filtered and concentrated. To the solution of this crude intermediate in THF (40 mL), was added tert-butyl 4- (2-aminoethyl)-l-piperazinecarboxylate (1.53 g, 1.1 eq., 6.68 mmol) and the mixture was allowed to stir at room temperature for 16 h. Progress of the reaction was monitored byELSD / TLC (SM was consumed). The reaction mixture was quenched with water (50 mL) and extracted with EtOAc (2x 30 mL). The separated organic layer was dried over Na2SO4and concentrated under reduced pressure. The resulting crude was purified over silica using 5% ethyl acetate in n-hexane to obtain [16E] (3 g, 54.04%) as a colourless oil.Results:

[0530] ELSD analysis: Purity 95.49 %, Calculated Formula: C50H104N4O5Si2 = 912.75, Observed =913.90 (m / z, M+H+).Intermediate (17E1,., OTBDMS7 Hk O,hkN‘ Y N‘•X.. J O kv, NHOTBDMS

[0531] As depicted in Scheme 14: To a stirred solution of [16E] (3 g, 3.28 mmol) in dichloromethane (50 mL), was added trifluoroacetic acid (2.51 mL, 10 eq., 32.8 mmol) at 0°C. The reaction mass was allowed to stir at RT for 4h. Progress of the reaction was monitored by TLC (SM was consumed). The reaction mass was concentrated under reduced pressure. The residue was dissolved in DCM (100 mL) and washed with saturated aq. solution of NaHCC>3(3x 25 mL) followed by water (2x 25 mL). The organic layer was dried over Na2SO4and concentrated under reduced pressure. The resulting crude [17E] (2.1 g, 78.62%) was forwarded to the next step without further purification.Results:

[0532] ELSD analysis: Purity 92.07 %, Calculated Formula: C45H96N4O4Si2 = 812.70, Observed =813.75 (m / z, M+H+).Intermediate [19E]Br'SCPh3

[0533] As depicted in Scheme 14: To a stirred solution of triphenylmethanethiol (25 g, 90.4 mmol) in ethanol (0.2 L) and water (0.2 L), was added sodium hydroxide (7.24 g, 2 eq., 181 mmol). After addition the reaction mass was left for 30 min at room temperature, then 1,2-dibromoethane (25.5 g, 1.5 eq., 136 mmol) was added dropwise. The reaction mixture was allowed to stir atroom temperature for 16 h. Reaction progress was monitored by TLC (after SM consumed). The reaction mixture was evaporated to remove excess ethanol. The residue was diluted with DCM (100 ml) and washed with water (2x 50 ml). The organic layer was dried over anhydrous sodium sulfate and distilled out under reduced pressure. The crude was crystalized in methanol to afford [19E] (30 g, 86.52%) as an off white solid.Results:

[0534] NMR (400 MHz, CDCl3): δ 7.50-7.43 (d, 7 = 7.52 Hz, 6H), 7.36-7.23 (m, 9H), 2.89 (t, J = 7.56 Hz, 2H), 2.74 (t, J = 8.52 Hz, 2H).Intermediate (20E1SCPh3OTBDMS

[0535] As depicted in Scheme 14: To a stirred solution of [17E] (2.1 g, 2.58 mmol) in dimethylformamide (40 mL) was added dipotassium carbonate (1.07 g, 3 eq., 7.74 mmol) followed by addition of [19E] (1.19 g, 1.2 eq., 3.1 mmol). The reaction mass was allowed to stir at room temperature for 16 h. Progress of the reaction was monitored by ELSD / TLC (SM was consumed). The reaction mixture was quenched with water (100 mL) and extracted with diethyl ether (2x 30 mL). The separated organic layer was dried over Na2SO4and concentrated under reduced pressure. The resulting crude was purified over silica using 5% MeOH in DCM to obtain [20E] (2.1 g, 72.9%) as a pale yellow oil.Results:

[0536] ELSD analysis: Purity 90.95 %, Calculated Formula: C66H114N4O4SSi2= 1114.81, Observed = 1116.15 (m / z, M+H+).Intermediate (21E1OTBDMS

[0537] As depicted in Scheme 14: To a stirred solution of [20E] (2.3 g, 2.06 mmol) in dichloromethane (16 mL), were added triethylsilane (396 pL, 1.2 eq., 2.47 mmol) and trifluoroacetic acid (3.94 mL, 25 eq., 51.5 mmol) simultaneously at 0°C under a nitrogen atmosphere. The reaction mass was allowed to stir for 5 min at the same temperature. Progress of the reaction was monitored by TLC (SM was consumed). The reaction mass was concentrated under reduced pressure. The residue was dissolved in DCM (100 mL) and washed with aq. saturated solution of NaHCO₃ (2x 20 mL) and water (2x 20 mL). The organic layer was dried over Na2SO4and concentrated under reduced pressure to obtain [21E] 2.3 g (crude), which was forwarded to the next step without further purification.Results:

[0538] ELSD analysis: Purity 48.66%, Calculated Formula: C47H100N4O4SSi2= 872.70, Observed = 873.05 (m / z, M+H+).Intermediate [22E]OTBDMS

[0539] As depicted in Scheme 14: A stirred solution of [21E] (1.8 g, 2.06 mmol) and [9E] (1.56 g, 2.06 mmol) in dichloromethane (16 mL) was allowed to stir at RT for 16h. Progress of the reaction was monitored by TLC / ELSD. The reaction mixture was concentrated under reduced pressure. The resulting crude was purified on a silica gel flash column by using 0-20 % EtOAc in Heptane to afford [22E] (1.2 g, 38.36%) as a colourless oil.Results:

[0540] ELSD analysis: Purity 90.50 %, Calculated Formula: C83H177N5O6S2Si4= 1516.22, Observed = 1517.95 (m / z, M+H+).Compound 43OH

[0541] As depicted in Scheme 14: To a stirred solution of [22E] (0.5 g, 329 pmol) in tetrahydrofuran (20 mL) was added 1-pyridinium fluoride (326 mg, 10 eq., 3.29 mmol) dropwise at 0°C, then the reaction mixture was brought to room temperature and stirred for 16 h. Reaction progress was monitored by TLC / ELSD. The reaction mass was diluted with diethyl ether (25 mL) and washed with water (2x 10 mL). T...

Claims

CLAIMS1. A compound having a structure according to Formula (I):or a pharmaceutically acceptable salt thereof, wherein:H NA1is selected from a bond,, -O-, -S-, and -(C1-C6alkylene)-, wherein the left hand side of each depicted structure is bound to the -(CH2)a-;wherein the right hand side of each depicted structure is bound to the -(CH2)a-;each R is independently selected from:o" r'-' Ojr(i)d, wherein each R1is independently selected from optionally substituted alkyl, optionally substituted alkenyl, and optionally substituted alkynyl;(ii) OH, wherein each R2is independently selected from optionally substituted alkyl, optionally substituted alkenyl, optionally substituted alkynyl, and -W1-X1, wherein each W1is independently selected from optionally substituted alkylene and optionally substituted alkenylene, andeach X1is independently selected from -*O-(C=O)-optionally substituted alkyl, - (*C=O)-O-optionally substituted alkyl, -*O-(C=O)-optionally substituted alkenyl, and - (*C=O)-O-optionally substituted alkenyl, wherein the atom marked with a * is connected to W1;(iii), wherein each R3is independently selected from optionally substituted alkyl, optionally substituted alkenyl, and optionally substituted alkynyl; and(iv)SIT, wherein each R4is independently selected from optionally substituted alkyl, optionally substituted alkenyl, and optionally substituted alkynyl;(v) optionally substituted alkyl or optionally substituted alkenyl;each R5is selected from hydrogen and optionally substituted (Ci-Cs) alkyl;each R6is selected from optionally substituted (Ci-Cs) alkyl;each a is independently selected from 1, 2, 3, 4, 5, 6, 7, 8, 9 and 10;each b is 1, 2, 3, 4 or 5;each c is 1, 2, 3, 4, or 5;each d is independently selected from 2, 3, 4, 5, 6, 7, 8, 9 and 10;each e is independently selected from 2, 3, 4, 5, 6, 7, 8, 9 and 10; andeach f is independently selected from 2, 3, 4, 5, 6, 7, 8, 9 and 10.

2. The compound of claim 1, wherein the compound has a structure according to Formula (IC):(IC)or a pharmaceutically acceptable salt thereof, wherein each R2A, R2B, R2Cand R2Dis independently selected from optionally substituted alkyl, optionally substituted alkenyl, optionally substituted alkynyl, and -W1-X1,wherein each W1is independently selected from optionally substituted alkylene and optionally substituted alkenylene, andeach X1is independently selected from -*O-(C=O)-optionally substituted alkyl, -(*C=O)-O-optionally substituted alkyl, -*O-(C=O)-optionally substituted alkenyl, and -(*C=O)-O-optionally substituted alkenyl, wherein the atom marked with a * is connected to W1.

3. The compound of claim 1, wherein the compound has a structure according to Formula (ID):(ID) or a pharmaceutically acceptable salt thereof, wherein each R2A, R2B, R2Cand R2Dis independently selected from optionally substituted alkyl, optionally substituted alkenyl, optionally substituted alkynyl, and -W1-X1,wherein each W1is independently selected from optionally substituted alkylene and optionally substituted alkenylene, andeach X1is independently selected from -*O-(C=O)-optionally substituted alkyl, -(*C=O)-O-optionally substituted alkyl, -*O-(C=O)-optionally substituted alkenyl, and -(*C=O)-O-optionally substituted alkenyl, wherein the atom marked with a * is connected to W1.

4. The compound of any one of the preceding claims, or a pharmaceutically acceptable salt thereof, wherein each a is independently selected from 2, 3 and 4.

5. The compound of any one of the preceding claims, or a pharmaceutically acceptable salt thereof, wherein b is selected from 2, 3 and 4.

6. The compound of any one of the preceding claims, or a pharmaceutically acceptable salt thereof, wherein c is selected from 2, 3 and 4.

7. The compound of any one of the preceding claims, or a pharmaceutically acceptable salt thereof, wherein Z1is -S-S-.

8. The compound of any one of the preceding claims, or a pharmaceutically acceptable salt thereof, wherein(a) R5and R6are both methyl; or(b) R5is hydrogen and R6is methyl.

9. A compound selected from those listed in Table 1, or a pharmaceutically acceptable salt thereof.

10. A composition comprising the compound of any one of the preceding claims, or a pharmaceutically acceptable salt thereof, and further comprising:(i) one or more non-cationic lipids,(ii) one or more cholesterol-based lipids, and(iii) one or more PEG-modified lipids.

11. The composition of claim 10, wherein the composition is a lipid nanoparticle, optionally a liposome.

12. The composition of claim 11, wherein(a) the one or more cationic lipid(s) constitute(s) about 30 mol %-60 mol % of the lipid nanoparticle;(b) the one or more non-cationic lipid(s) constitute(s) about 10 mol %-50 mol % of the lipid nanoparticle;(c) the one or more PEG-modified lipid(s) constitute(s) about 1 mol %-10 mol % of the lipid nanoparticle; and / or(d) the cholesterol-based lipid constitutes about 10 mol %-50 mol% of the lipid nanoparticle.

13. The composition of claim 11 or 12, wherein the lipid nanoparticle encapsulates an mRNA encoding a peptide or protein, optionally for use in a vaccine, optionally wherein the lipid nanoparticles have an encapsulation percentage for mRNA of(i) at least 50%;(ii) at least 55%;(iii) at least 60%;(iv) at least 65%;(v) at least 70%;(vi) at least 75%;(vii) at least 80%;(viii) at least 85%;(ix) at least 90%; or(x) at least 95%.

14. A method for treating, preventing or ameliorating a disease, disorder or infection, wherein said method comprises administering to a subject in need thereof the composition of claim 13 and wherein the disease, disorder or infection is amenable to treatment or prevention by the peptide or protein encoded by the mRNA, optionally wherein the disease is (a) a protein deficiency, optionally wherein the protein deficiency affects the liver, lung, brain or muscle, (b) an autoimmune disease, (c) an infectious disease, or (d) cancer, optionally wherein the composition is administered intranasally, intravenously, intrathecally or intramuscularly, or by pulmonary delivery, optionally through nebulization, for example wherein the composition is administered:(i) intramuscularly, or(ii) intravenously.

15. A method for inducing an immune response in a subject wherein said method comprises administering to a subject in need thereof the composition of claim 13, optionally wherein the composition is administered intranasally, intravenously, intrathecally or intramuscularly, or by pulmonary delivery, optionally through nebulization, for example wherein the composition is administered:(i) intramuscularly, or(ii) intravenously.

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