Cationic lipids based on phenolic acid lipids

Abstract

The present invention provides, in part, phenolic acid lipid compounds of Formula (I) and their sub-formulae or pharmaceutically acceptable salts thereof. The compounds provided herein are useful for the delivery and expression of mRNA and encoded proteins, e.g., as components of liposomal delivery vehicles, and are thus useful in the treatment of various diseases, disorders and conditions, such as diseases, disorders and conditions associated with the deficiency of one or more proteins.

Description

[0001] This application is a divisional application of Chinese patent application No. 202180039080.X, entitled "Catonic Lipids Based on Phenolic Acid Lipids", filed on March 31, 2021 (PCT application No. PCT / US2021 / 025128).

[0002] Cross-referencing related applications

[0003] This application claims priority to U.S. Provisional Patent Application 63 / 003,698, filed April 1, 2020, which is incorporated herein by reference in its entirety. Background Technology

[0004] Nucleic acid delivery has been extensively explored as a potential therapeutic option for certain disease states. Specifically, messenger RNA (mRNA) therapy has become an increasingly important option for treating a variety of diseases, including those associated with the deficiency of one or more proteins.

[0005] Efficient delivery of nucleic acids encapsulated in liposomes remains an active area of ​​research. Cationic lipid components play a crucial role in facilitating the efficient encapsulation of nucleic acids during liposome loading. Furthermore, cationic lipids can play a vital role in the efficient release of nucleic acid cargo from liposomes into the cytoplasm of target cells. Various cationic lipids suitable for in vivo use have been identified. However, there is still a need to identify lipids that can be synthesized efficiently and inexpensively without forming potentially toxic byproducts.

[0006] Phenolic acids possess many advantageous properties, making them a good starting point for the synthesis of cationic lipids for use in the in vivo environment. For example, phenolic acids have not shown toxicity, are readily available, and are easily derivatized. Broadly speaking, phenolic acids can be divided into two groups: benzoic acid and cinnamic acid, and their derivatives.

[0007] Examples of benzoic acid that can be used to synthesize the cationic lipids of the present invention include:

[0008]

[0009]

[0010] Examples of cinnamic acid that can be used to synthesize the cationic lipids of the present invention include:

[0011] In some embodiments, examples of cinnamic acid that can be used to synthesize the cationic lipids of the present invention include:

[0012]

[0013] In some embodiments, examples of cinnamic acid that can be used to synthesize the cationic lipids of the present invention include:

[0014]

[0015] In some embodiments, examples of cinnamic acid that can be used to synthesize the cationic lipids of the present invention include:

[0016]

[0017] In some embodiments, examples of cinnamic acid that can be used to synthesize the cationic lipids of the present invention include:

[0018] Summary of the Invention

[0019] Among other things, this invention provides a novel class of cationic lipid compounds for in vivo delivery of therapeutic agents such as nucleic acids. These compounds are expected to be delivered efficiently in vivo while maintaining favorable toxicity characteristics.

[0020] The cationic lipids of the present invention can be synthesized from readily available starting reagents such as phenolic acids, benzoic acid, and cinnamic acid. The cationic lipids of the present invention also exhibit unexpectedly high encapsulation efficiency. The cationic lipids of the present invention further include cleavable groups (e.g., esters and disulfides), which are considered to improve biodegradability and thus contribute to their advantageous toxicity characteristics.

[0021] On the one hand, cationic lipids having a structure according to formula (I) are provided:

[0022]

[0023] Where L1 is a bond, (C1-C6)alkyl or (C2-C6)alkenyl;

[0024] Where X is O or S;

[0025] Where R 1 R 2 R 3 R 4 and R 5 Each is independently selected from H, OH, optionally substituted (C1-C6)alkyl, optionally substituted (C2-C6)alkenyl, optionally substituted (C2-C6)ynyl, optionally substituted (C1-C6)alkoxy and -OC(O)R';

[0026] Where R 1 R 2 R 3 R 4 or R 5At least one of them is -OC(O)R';

[0027] Where R' is

[0028]

[0029] Where R 6 for

[0030]

[0031] Where m and p are each independently 0, 1, 2, 3, 4 or 5;

[0032] Where R 7 Selected from H, optionally substituted (C1-C6)alkyl, optionally substituted (C2-C6)alkenyl, optionally substituted (C2-C6)ynyl, optionally substituted (C1-C6)acyl, -(CH2). k R A Or -(CH2) k CH(OR 11 )R A ;

[0033] Where R 8 Selected from H, optionally substituted (C1-C6)alkyl, optionally substituted (C2-C6)alkenyl, optionally substituted (C2-C6)ynyl, optionally substituted (C1-C6)acyl, -(CH2). n R B Or -(CH2) n CH(OR 12 )R B ;

[0034] Where R 9 Selected from H, optionally substituted (C1-C6)alkyl, optionally substituted (C2-C6)alkenyl, optionally substituted (C2-C6)ynyl, optionally substituted (C1-C6)acyl, -(CH2). q R C Or -(CH2) q CH(OR 13 )R C ;

[0035] Where R 10 Selected from H, optionally substituted (C1-C6)alkyl, optionally substituted (C2-C6)alkenyl, optionally substituted (C2-C6)ynyl, optionally substituted (C1-C6)acyl, -(CH2). r R D Or -(CH2) r CH(OR 14 )R D ;

[0036] Where k, n, q and r are each 1, 2, 3, 4 or 5 independently;

[0037] Or one of them (i)R 7 and R 8 Or (ii)R 9 and R 10 Together they form optionally substituted 5- or 6-membered heterocyclic alkyl or heteroaryl groups, wherein the heterocyclic alkyl or heteroaryl group comprises 1 to 3 heteroatoms selected from N, O and S;

[0038] Where R 11 R 12 R 13 and R 14 Each is independently selected from H, methyl, ethyl, or propyl;

[0039] Where R A R B R C and R D Each is independently selected from the arbitrarily substituted (C6-C) 20 )alkyl, optionally substituted (C6-C 20 alkenyl, optionally substituted (C6-C) 20 ) ynyl group, optionally substituted (C6-C 20 Acyl, optionally substituted -OC(O)alkyl, optionally substituted -OC(O)alkenyl, optionally substituted (C1-C6)monoalkylamino, optionally substituted (C1-C6)dialkylamino, optionally substituted (C1-C6)alkoxy, -OH, -NH2;

[0040] Where R 7 R 8 R 9 R 10 At least one of them includes R A R B R C or R D Part, wherein the R A R B R C or R D Independently selected from optional substitutions (C6-C) 20 )alkyl, optionally substituted (C6-C 20 alkenyl, optionally substituted (C6-C) 20 ) ynyl group, optionally substituted (C6-C 20 Acyl group, optionally substituted -OC(O)(C6-C 20 )alkyl or optionally substituted -OC(O)(C6-C 20 alkenyl;

[0041] Or its pharmaceutically acceptable salt.

[0042] On the one hand, this article provides cationic lipids as pharmaceutically acceptable salts of formula (I).

[0043] On one hand, this document provides compositions comprising the cationic lipids of the present invention, one or more non-cationic lipids, one or more cholesterol-based lipids, and one or more PEG-modified lipids. On the other hand, said compositions are lipid nanoparticles, optionally liposomes.

[0044] On the one hand, the composition comprising the cationic lipids of the present invention can be used for therapeutic purposes. Attached Figure Description

[0045] Figure 1 In vivo protein expression following intratracheal administration of lipid nanoparticles comprising one of cationic lipid compounds 1-12 was demonstrated. Based on positive luciferase activity, lipid nanoparticles comprising the cationic lipids described herein efficiently delivered FFL mRNA in vivo. Detailed Implementation

[0046] definition

[0047] To facilitate understanding of the invention, certain terms are defined below. Further definitions of the following and other terms are set forth throughout this specification. Publications and other references cited herein to describe the background art and provide additional details about its implementation are incorporated herein by reference.

[0048] Amino acids: 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 a l-amino acid. "Standard amino acid" refers to any of the twenty standard I-amino acids commonly found in naturally occurring peptides. "Non-standard amino acid" refers to any amino acid other than a standard amino acid, whether it is synthetically prepared 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 (e.g., amides), and / or substitutes. Amino acids, including carboxyl 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 alter the cyclic half-life of the peptide without adversely affecting its activity. Amino acids can participate in disulfide bonds. Amino acids may contain one or more post-translational modifications, such as those associated with one or more chemical entities (e.g., methyl groups, acetate groups, acetyl groups, phosphate groups, formyl moieties, isoprene-like groups, sulfate groups, polyethylene glycol moieties, lipid moieties, carbohydrate moieties, and biotin moieties, etc.). The terms "amino acid" and "amino acid residue" are used interchangeably and can refer to free amino acids and / or amino acid residues of peptides. Whether the term refers to free amino acids or peptide residues will be apparent from the context in which the term is used.

[0049] Animal: As used herein, the term "animal" means any member of the animal kingdom. In some embodiments, "animal" means a human being at any developmental stage. In some embodiments, "animal" means a non-human animal at any developmental stage. In some embodiments, non-human animals are mammals (e.g., rodents, mice, rats, rabbits, monkeys, dogs, cats, sheep, cattle, primates, and / or pigs). In some embodiments, animals include, but are not limited to, mammals, birds, reptiles, amphibians, fish, insects, and / or worms. In some embodiments, animals may be transgenic animals, genetically engineered animals, and / or clones.

[0050] Approximately or about: As used herein, when applied to one or more target values, the term “approximately” or “about” refers to a value similar to the reference value. In some embodiments, the term “approximately” or “about” refers to a range of values ​​falling 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 value, unless otherwise stated or otherwise obvious from the context (unless the number exceeds 100% of the possible value).

[0051] Biologically active: As used herein, the term "biologically active" refers to the characteristic of any agent that is active in a biological system, particularly in an organism. For example, an agent that has a biological effect on an organism when applied to that organism is considered biologically active.

[0052] Delivery: As used herein, the term “delivery” encompasses both local delivery and systemic delivery. For example, mRNA delivery encompasses situations where the mRNA is delivered to a target tissue and the encoded protein is expressed and retained within the target tissue (also known as “local distribution” or “local delivery”), and situations where the mRNA is delivered to a target tissue and the encoded protein is expressed and secreted into the patient’s circulatory system (e.g., serum), and then distributed systemically and absorbed by other tissues (also known as “systemic distribution” or “systemic delivery”).

[0053] Expression: As used herein, “expression” of a nucleic acid sequence refers to post-translational modifications that translate mRNA into a polypeptide, assemble multiple polypeptides into a complete protein (e.g., an enzyme), and / or a polypeptide or a fully assembled protein (e.g., an enzyme). In this application, the terms “expression” and “production” and their grammatical equivalents are used interchangeably.

[0054] Functionality: As used herein, a “functional” biomolecule is a biomolecule that exhibits the form that characterizes its properties and / or activities.

[0055] Half-life: As used herein, the term “half-life” is the time required for the concentration or activity of a nucleic acid or protein to decrease to half of its value as measured at the beginning of a time period.

[0056] Supporting lipids: As used herein, the term "supporting lipid" refers to any neutral or zwitterionic lipid material containing cholesterol. Without being bound by any particular theory, supporting lipids can increase the stability, rigidity, and / or fluidity within lipid bilayers / nanoparticles.

[0057] Improvement, increase, or decrease: As used herein, the terms “improvement,” “increase,” or “decrease,” or their grammatical equivalents, refer to a value 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 subjects) in the absence of the treatment described herein. A “control individual” is an individual suffering from the same form of disease as the treated individual and of approximately the same age as the treated individual.

[0058] In vitro: As used herein, the term “in vitro” refers to events that occur in an artificial environment, such as in a test tube or reaction vessel, in a cell culture, etc., rather than in a multicellular organism.

[0059] In vivo: As used herein, the term "in vivo" refers to events occurring within multicellular organisms such as humans and non-human animals. In the context of cell-based systems, the term can be used to refer to events occurring within living cells (as opposed to, for example, in vitro systems).

[0060] Separated: As used herein, the term “separated” means (1) a substance and / or entity that is separated from at least some of the components that were originally associated with it at the time of its production (whether natural and / or in an experimental setting), and / or (2) a substance and / or entity that is artificially produced, prepared, and / or manufactured. Separated substances and / or entities may be separated from other components that were originally associated with them by about 10%, about 20%, about 30%, about 40%, about 50%, about 60%, about 70%, about 80%, about 90%, about 90%, about 91%, about 92%, about 93%, about 94%, about 95%, about 96%, about 97%, about 98%, about 99%, or more than about 99%. In some embodiments, the purity of the separated reagent is about 80%, about 85%, about 90%, about 91%, about 92%, about 93%, about 94%, about 95%, about 96%, about 97%, about 98%, about 99%, or more than 99%. As used herein, a substance is "pure" if it is substantially free of other components. As used herein, the calculation of the purity percentage of the separated substance and / or entity should not include excipients (e.g., buffers, solvents, water, etc.).

[0061] Liposomes: As used herein, the term "liposome" refers to any layered, multilayered, or solid nanoparticle vesicle. Typically, as used herein, liposomes can be formed by mixing one or more lipids or by mixing one or more lipids and polymers. In some embodiments, liposomes suitable for use in the present invention contain one or more cationic lipids and optionally one or more non-cationic lipids, optionally one or more cholesterol-based lipids, and / or optionally one or more PEG-modified lipids.

[0062] Messenger RNA (mRNA): As used herein, the term “messenger RNA (mRNA)” or “mRNA” refers to a polynucleotide encoding at least one polypeptide. As used herein, mRNA includes modified RNA and unmodified RNA. The term “modified mRNA” refers to mRNA containing at least one chemically modified nucleotide. mRNA may contain one or more coding and noncoding regions. mRNA may be purified from natural sources, produced using recombinant expression systems, and optionally purified, chemically synthesized, etc. Where appropriate, such as in the case of chemically synthesized molecules, mRNA may contain nucleoside analogs, such as analogs with chemically modified bases or sugars, backbone modifications, etc. Unless otherwise stated, the mRNA sequence is displayed in the 5' to 3' orientation. In some embodiments, the mRNA is or comprises a natural nucleoside (e.g., adenosine, guanosine, cytidine, uridine); or a nucleoside analogue (e.g., 2-aminoadenosine, 2-thiothymidine, inosine, pyrrolopyrimidine, 3-methyladenosine, 5-methylcytidine, C-5-propynylcytidine, C-5-propynyl-uridine, 2-aminoadenosine, C5-bromouridine, C5-fluorouridine, C5-iodouridine, C5-propynyluridine, C5-propynylcytidine, C5-methyl ...propynylcytidine, C5-methyluridine, C5-propynylcytidine, C5-propynylcytidine, C5-propynylcytidine, C5-propynylcytidine, C5-propynylcytidine, C5-propynylcytidine, C5-propynylcytidine, C5-propynylcytidine, C5-propynylcytidine, C5-propynylcytidine, C5-propynylcytidine, C5-propynylcytidine, C5-propynylcytidine, C5-propynylcytidine, C5-propynylcytidine Cytidine, 2-aminoadenosine, 7-deadenosine, 7-deadenosine, 8-oxoadenosine, 8-oxoguanosine, O(6)-methylguanosine and 2-thiocytidine); chemically modified bases; biologically modified bases (e.g., methylated bases); inserted bases; modified sugars (e.g., 2'-fluororibose, ribose, 2'-deoxyribose, arabinose and hexose); and / or modified phosphate groups (e.g., thiophosphates and 5'-N-phosphoramide bonds).

[0063] Nucleic acid: As used herein, the term “nucleic acid” in its broadest sense refers to any compound and / or substance that can be incorporated into a polynucleotide chain. In some embodiments, nucleic acid is a compound and / or substance incorporated into or that can be incorporated into a polynucleotide chain by a phosphodiester bond. In some embodiments, “nucleic acid” refers to a single nucleic acid residue (e.g., nucleotide and / or nucleoside). In some embodiments, “nucleic acid” refers to a polynucleotide chain containing a single nucleic acid residue. In some embodiments, “nucleic acid” encompasses RNA as well as single-stranded and / or double-stranded DNA and / or cDNA. In some embodiments, “nucleic acid” encompasses ribonucleic acid (RNA), including but not limited to any one or more of interfering RNA (RNAi), small interfering RNA (siRNA), short hairpin RNA (shRNA), antisense RNA (aRNA), messenger RNA (mRNA), modified messenger RNA (mmRNA), long non-coding RNA (lncRNA), microRNA (miRNA), polynucleotide coding (MCNA), polymeric nucleotide coding (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, a portion of plasmid DNA, pre-condensed DNA, a polymerase chain reaction (PCR) product, a vector (e.g., P1, PAC, BAC, YAC, artificial chromosome), an expression cassette, a chimeric sequence, chromosomal DNA, or derivatives of these groups.In embodiments, RNA may be messenger RNA (mRNA), ribosomal RNA (rRNA), signal recognition particle RNA (7SL RNA or SRP RNA), transfer RNA (tRNA), transfer messenger RNA (tmRNA), small nucleoRNA (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), splicing leader RNA (SL RNA), antisense RNA (aRNA or asRNA), cis-natural antisense transcript (cis-NAT), CRISPR RNA (crRNA), long noncoding RNA (lncRNA), microRNA (miRNA), piwi-interacting RNA (piRNA), small interfering RNA (siRNA), transaction siRNA (tasiRNA), repeat-associated siRNA (rasiRNA), 73K RNA, retrotransposons, viral genomes, viroids, satellite RNA, or derivatives of these groups. In some embodiments, nucleic acid is mRNA encoding proteins such as enzymes.

[0064] Patient: As used herein, the terms "patient" or "subject" refer to any organism to which the provided composition may be applied, for example, for experimental, diagnostic, preventative, 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, the patient is a human. Humans include prenatal and postnatal forms.

[0065] Pharmaceutically acceptable: As used herein, the term “pharmaceutically acceptable” means a substance that, in accordance with reasonable benefit / risk ratios and within the bounds of reasonable medical judgment, is suitable for contact with tissues in humans and animals without excessive toxicity, irritation, allergic reactions, or other problems or complications.

[0066] Pharmaceutically acceptable salts: Pharmaceutically acceptable salts are well known in the art. For example, SMBerge et al. described pharmaceutically acceptable salts in detail in the *Journal of Pharmaceutical Sciences*, (1977) 66:1-19. Pharmaceutically acceptable salts of the compounds of the present invention include those derived from suitable inorganic and organic acids and bases. Examples of pharmaceutically acceptable, non-toxic acid addition salts are those 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 formed by other methods used in the art, such as ion exchange, and containing an amino group. Other pharmaceutically acceptable salts include adipate, alginate, ascorbate, aspartate, benzenesulfonate, benzoate, hydrogen sulfate, borate, butyrate, camphorate, camphorsulfonate, citrate, cyclopentanepropionate, digluconate, dodecyl sulfate, ethanesulfonate, formate, fumarate, glucono-heptate, glyceryl phosphate, gluconate, hemisulfate, heptaate, hydroiodate, 2-hydroxyethanesulfonate, lactobionate, lactate, laurate, lauryl sulfate, malate, maleate, malonate, methanesulfonate, 2-naphthalenesulfonate, nicotinate, nitrate, oleate, oxalate, palmitate, dihydroxynaphthalate, pectate, persulfate, 3-phenylpropionate, phosphate, picrate, neopentanoate, propionate, stearate, succinate, sulfate, tartrate, thiocyanate, p-toluenesulfonate, undecanoate, valerate, etc. Salts derived from suitable bases include alkali metal salts, alkaline earth metal salts, ammonium salts, and N salts. + (C 1-4 Alkyl)4 salts. Representative alkali metal or alkaline earth metal salts include sodium, lithium, potassium, calcium, and magnesium salts. Where appropriate, other pharmaceutically acceptable salts include non-toxic ammonium, quaternary ammonium, and amine cations formed using counterions such as halides, hydroxides, carboxylates, sulfates, phosphates, nitrates, sulfonates, and arylsulfonates. Other pharmaceutically acceptable salts include those formed by the quaternization of amines using suitable electrophilic agents (e.g., alkyl halides) to form quaternized alkylated amino salts.

[0067] Whole-body distribution or delivery: As used herein, the terms “whole-body distribution” or “whole-body delivery” or their grammatical equivalents refer to a mechanism or method of delivery or distribution that affects the whole body or the entire organism. Generally, whole-body distribution or delivery is accomplished via the body’s circulatory system (e.g., blood flow). This contrasts with the definition of “local distribution or delivery.”

[0068] Subject: As used herein, the term “subject” refers to a human or any non-human animal (e.g., mouse, rat, rabbit, dog, cat, cow, pig, sheep, horse, or primate). Human includes both prenatal and postnatal forms. In many embodiments, the individual is a person. An individual can be a patient, which is a person who refers to a healthcare provider for diagnosis or treatment of a disease. The term “individual” is used interchangeably with “person” or “patient” herein. An individual may have or be susceptible to a disease or disorder, but may or may not exhibit symptoms of that disease or disorder.

[0069] Essentially: As used herein, the term "essentially" refers to qualitative conditions that exhibit all or nearly all of the target features or characteristics of a range or degree. Those skilled in the art of biology will understand that biological and chemical phenomena rarely (if ever) complete and / or continue to complete or achieve or avoid absolute results. Therefore, the term "essentially" is used herein to capture the inherent lack of completeness in many biological and chemical phenomena.

[0070] Target tissue: As used herein, the term “target tissue” refers to any tissue affected by the disease to be treated. In some embodiments, target tissue includes those tissues that exhibit disease-related pathology, symptoms, or features.

[0071] Therapeutic effective amount: As used herein, the term "therapeutic effective amount" means an amount sufficient to treat, diagnose, prevent, and / or delay the onset of symptoms of a disease, disorder, and / or condition when administered to a subject who has a disease, disorder, and / or condition or is susceptible to such disease, disorder, and / or condition. Those skilled in the art will recognize that a therapeutic effective amount is typically administered via a dosing regimen comprising at least one unit dose.

[0072] Treatment: As used herein, the term "treatment" means any method used to partially or completely alleviate, improve, reduce, suppress, prevent, delay the onset of, reduce the severity of, and / or decrease the incidence of one or more symptoms or features of a particular disease, disorder, and / or condition. Treatment may be administered to subjects who do not exhibit signs of disease and / or only exhibit early signs of disease in order to reduce the risk of developing a pathology associated with the disease.

[0073] Chemical definition

[0074] Acyl group: As used herein, the term "acyl group" refers to R Z -(C=O)-, where R Z For example, any alkyl, alkenyl, ynyl, heteroalkyl, or heteroalkylene.

[0075] Aliphatic: As used in this article, the term aliphatic refers to C 1- C 40Hydrocarbons, including both saturated and unsaturated hydrocarbons. Aliphatic hydrocarbons can be straight-chain, branched, or cyclic. For example, C1-C 20 Aliphatic can include C1-C 20 Alkyl groups (e.g., straight-chain or branched C1-C) 20 saturated alkyl), C2-C 20 Alkenyl (e.g., straight-chain or branched C4-C) 20 Diene-based, straight-chain or branched C6-C 20 Trienyl groups, etc., and C2-C 20 Alkyne groups (e.g., straight-chain or branched C2-C) 20 Alkyne group). C1-C 20 Aliphatic bacteria can include C3-C 20 Cyclic aliphatic (e.g., C3-C) 20 cycloalkyl, C4-C 20 Cycloalkenyl or C8-C 20 (Cycloalkyne). In some embodiments, the aliphatic group may comprise one or more cyclic aliphatic groups 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, halogen, alkoxy, hydroxyl, amino, aryl, ether, ester, or amide. The aliphatic group is unsubstituted or substituted with one or more substituents as described herein. For example, the aliphatic group may be substituted with one or more of the following (e.g., 1, 2, 3, 4, 5, or 6 independently selected substituents): halogen, -COR", -CO2H, -CO2R", -CN, -OH, -OR", -OCOR', -OCO2R", -NH2, -NHR", -N(R")2, -SR", or -SO2R", wherein each instance of R" is independently C1-C1. 20 Aliphatic (e.g., C1-C) 20 Alkyl, C1-C 15 Alkyl, C1-C 10 Alkyl or C1-C3 alkyl). In the examples, R” is independently an unsubstituted alkyl group (e.g., unsubstituted C1-C3 alkyl). 20 Alkyl, C1-C 15 Alkyl, C1-C 10 Alkyl or C1-C3 alkyl). In the examples, "R" is independently an unsubstituted C1-C3 alkyl. In the examples, aliphatic is unsubstituted. In the examples, aliphatic does not include any heteroatoms. Alkyl: As used herein, the term "alkyl" means a noncyclic straight-chain and branched hydrocarbon group, such as "C1-C3 alkyl". 30"Alkyl" refers to an alkyl group having 1 to 30 carbon atoms. Alkyl groups can be straight-chain 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-pentyl, hexyl, isohexyl, etc. The term "lower alkyl" refers to a straight-chain or branched alkyl group having 1 to 6 carbon atoms. Other alkyl groups will be apparent to those skilled in the art in light of the beneficial effects of this disclosure. Alkyl groups can be unsubstituted or substituted with one or more substituents as described herein. For example, an alkyl group can be substituted with one or more of the following (e.g., 1, 2, 3, 4, 5, or 6 independently selected substituents): halogen, -COR", -CO2H, -CO2R", -CN, -OH, -OR", -OCOR', -OCO2R", -NH2, -NHR", -N(R")2, -SR", or -SO2R", wherein each instance of R" is independently C1-C2. 20 Aliphatic (e.g., C1-C) 20 Alkyl, C1-C 15 Alkyl, C1-C 10 Alkyl or C1-C3 alkyl). In the examples, R” is independently an unsubstituted alkyl group (e.g., unsubstituted C1-C3 alkyl). 20 Alkyl, C1-C 15 Alkyl, C1-C 10 Alkyl or C1-C3 alkyl). In the examples, "R" is independently an unsubstituted C1-C3 alkyl. In the examples, the alkyl group is substituted (e.g., substituted by 1, 2, 3, 4, 5, or 6 substituent groups as described herein). In the examples, the alkyl group is substituted with a -OH group and may also be referred to herein as "hydroxyalkyl", wherein the prefix indicates a -OH group, and "alkyl" is as described herein.

[0076] As used herein, the term "alkyl" also refers to a free radical ("C1-C50") of a straight-chain or branched saturated hydrocarbon group having 1 to 50 carbon atoms. 50 Alkyl group (“alkyl”). In some embodiments, the alkyl group has 1 to 40 carbon atoms (“C1-C4”). 40 Alkyl group (“alkyl”). In some embodiments, the alkyl group has 1 to 30 carbon atoms (“C1-C3”). 30 Alkyl group (“alkyl”). In some embodiments, the alkyl group has 1 to 20 carbon atoms (“C1-C2”). 20 Alkyl group (“alkyl”). In some embodiments, the alkyl group has 1 to 10 carbon atoms (“C1-C1”). 10The alkyl group (“C1-C9 alkyl”) has 1 to 9 carbon atoms in some embodiments. In some embodiments, the alkyl group has 1 to 8 carbon atoms (“C1-C8 alkyl”). In some embodiments, the alkyl group has 1 to 7 carbon atoms (“C1-C7 alkyl”). In some embodiments, the alkyl group has 1 to 6 carbon atoms (“C1-C6 alkyl”). In some embodiments, the alkyl group has 1 to 5 carbon atoms (“C1-C5 alkyl”). In some embodiments, the alkyl group has 1 to 4 carbon atoms (“C1-C4 alkyl”). In some embodiments, the alkyl group has 1 to 3 carbon atoms (“C1-C3 alkyl”). In some embodiments, the alkyl group has 1 to 2 carbon atoms (“C1-C2 alkyl”). In some embodiments, the alkyl group has 1 carbon atom (“C1 alkyl”). In some embodiments, the alkyl group has 2 to 6 carbon atoms (“C2-C6 alkyl”). Examples of C1-C6 alkyl groups include, but are not limited to, methyl (C1), ethyl (C2), n-propyl (C3), isopropyl (C3), n-butyl (C4), tert-butyl (C4), sec-butyl (C4), isobutyl (C4), n-pentyl (C5), 3-pentyl (C5), pentyl (C5), neopentyl (C5), 3-methyl-2-butyl (C5), tert-pentyl (C5), and n-hexyl (C6). Further examples of alkyl groups include n-heptyl (C7), n-octyl (C8), etc. Unless otherwise stated, each example of an alkyl group is independently unsubstituted (“unsubstituted alkyl”) or substituted with one or more substituents (“substituted alkyl”). In some embodiments, the alkyl group is an unsubstituted C1-C6 alkyl group. 50 Alkyl group. In some embodiments, the alkyl group is a substituted C1-C alkyl group. 50 alkyl.

[0077] Adding the prefix "a" to a group indicates that the group is a divalent moiety. For example, arylene is a divalent moiety of aryl, and heteroarylene is a divalent moiety of heteroaryl.

[0078] Alkylene: As used herein, the term "alkylene" refers to a saturated divalent straight-chain or branched hydrocarbon group, and exemplified by methylene, ethylene, isopropylene, etc. Similarly, as used herein, the term "alkenylene" refers to an unsaturated divalent straight-chain or branched hydrocarbon group having one or more unsaturated carbon-carbon double bonds, which may be present at any stable point along the chain, and the term "alkynylene" herein refers to an unsaturated divalent straight-chain or branched hydrocarbon group having one or more unsaturated carbon-carbon triple bonds, which may be present at any stable point along the chain. In some embodiments, alkylene, alkenylene, or alkynylene 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, halogen, alkoxy, hydroxyl, amino, aryl, ether, ester, or amide. For example, an alkylene, alkenylene, or ynylene group may be substituted with one or more of the following (e.g., 1, 2, 3, 4, 5, or 6 independently selected substituents): halogen, -COR", -CO2H, -CO2R", -CN, -OH, -OR", -OCOR", -OCO2R", -NH2, -NHR", -N(R")2, -SR", or -SO2R", wherein each instance of R" is independently C1-C2. 20 Aliphatic (e.g., C1-C) 20 Alkyl, C1-C 15 Alkyl, C1-C 10 Alkyl or C1-C3 alkyl). In the examples, R” is independently an unsubstituted alkyl group (e.g., unsubstituted C1-C3 alkyl). 20 Alkyl, C1-C 15 Alkyl, C1-C 10 Alkyl or C1-C3 alkyl). In the examples, "R" is independently an unsubstituted C1-C3 alkyl. In some examples, the alkylene, alkenyl, or ynylene is unsubstituted. In some examples, the alkylene, alkenyl, or ynylene does not include any heteroatoms. Alkenyl: As used herein, "alkenyl" means any straight or branched hydrocarbon chain having one or more unsaturated carbon-carbon double bonds, which may appear at any stable point along the chain, for example: "C2-C 30"Alkenyl" refers to an alkenyl group having 2-30 carbon atoms. Examples of alkenyl groups include prop-2-enyl, but-2-enyl, but-3-enyl, 2-methylprop-2-enyl, hex-2-enyl, hex-5-enyl, 2,3-dimethylbut-2-enyl, etc. In embodiments, the alkenyl group comprises 1, 2, or 3 carbon-carbon double bonds. In embodiments, the alkenyl group comprises a single carbon-carbon double bond. In embodiments, multiple double bonds (e.g., 2 or 3) are conjugated. The alkenyl group may be unsubstituted or substituted with one carbon atom. Or multiple substituents described herein. For example, the alkenyl group may be substituted by one or more of the following (e.g., 1, 2, 3, 4, 5 or 6 independently selected substituents): halogen, -COR", -CO2H, -CO2R", -CN, -OH, -OR", -OCOR", -OCO2R", -NH2, -NHR", -N(R")2, -SR" or -SO2R", wherein each instance of R" is independently C1-C 20 Aliphatic (e.g., C1-C) 20 Alkyl, C1-C 15 Alkyl, C1-C 10 Alkyl or C1-C3 alkyl). In the examples, R” is independently an unsubstituted alkyl group (e.g., unsubstituted C1-C3 alkyl). 20 Alkyl, C1-C 15 Alkyl, C1-C 10 Alkyl or C1-C3 alkyl). In the examples, "R" is independently an unsubstituted C1-C3 alkyl. In the examples, the alkenyl group is unsubstituted. In the examples, the alkenyl group is substituted (e.g., substituted by 1, 2, 3, 4, 5, or 6 substituent groups as described herein). In the examples, the alkenyl group is substituted with a -OH group and may also be referred to herein as "hydroxyalkenyl," where the prefix indicates a -OH group, and "alkenyl" is as described herein.

[0079] As used herein, “alkenyl” also refers to a free radical (“C2-C”) of a straight-chain or branched hydrocarbon group having 2 to 50 carbon atoms and one or more carbon-carbon double bonds (e.g., 1, 2, 3 or 4 double bonds). 50 Alkenyl group (“Alkenyl”). In some embodiments, the alkenyl group has 2 to 40 carbon atoms (“C2-C”). 40 Alkenyl group (“Alkenyl”). In some embodiments, the alkenyl group has 2 to 30 carbon atoms (“C2-C”). 30 Alkenyl group (“Alkenyl”). In some embodiments, the alkenyl group has 2 to 20 carbon atoms (“C2-C”). 20 Alkenyl group (“Alkenyl”). In some embodiments, the alkenyl group has 2 to 10 carbon atoms (“C2-C”). 10The alkenyl group is referred to as "alkenyl". In some embodiments, the alkenyl group has 2 to 9 carbon atoms ("C2-C9 alkenyl"). In some embodiments, the alkenyl group has 2 to 8 carbon atoms ("C2-C8 alkenyl"). In some embodiments, the alkenyl group has 2 to 7 carbon atoms ("C2-C7 alkenyl"). In some embodiments, the alkenyl group has 2 to 6 carbon atoms ("C2-C6 alkenyl"). In some embodiments, the alkenyl group has 2 to 5 carbon atoms ("C2-C5 alkenyl"). In some embodiments, the alkenyl group has 2 to 4 carbon atoms ("C2-C4 alkenyl"). In some embodiments, the alkenyl group has 2 to 3 carbon atoms ("C2-C3 alkenyl"). In some embodiments, the alkenyl group has 2 carbon atoms ("C2 alkenyl"). One or more carbon-carbon double bonds may be located internally. Examples of C2-C4 alkenyl groups include, but are not limited to, vinyl (C2), 1-propenyl (C3), 2-propenyl (C3), 1-butenyl (C4), 2-butenyl (C4), butadienyl (C4), etc. Examples of C2-C6 alkenyl groups include the aforementioned C2-C4 alkenyl groups as well as pentenyl (C5), pentadienyl (C5), hexenyl (C6), etc. Other examples of alkenyl groups include heptenyl (C7), octenyl (C8), octtrienyl (C8), etc. Unless otherwise stated, each example of an alkenyl group is independently unsubstituted (“unsubstituted alkenyl”) or substituted with one or more substituents (“substituted alkenyl”). In some embodiments, the alkenyl group is an unsubstituted C2-C4 alkenyl group. 50 Alkenyl group. In some embodiments, the alkenyl group is a substituted C2-C group. 50 Alkenyl group.

[0080] Alkynyl: As used herein, “alkynyl” means any straight-chain or branched hydrocarbon chain having one or more carbon-carbon triple bonds present at any stable point along the chain, such as “C2-C”. 30 "Alynyl" refers to an alkynyl group having 2-30 carbon atoms. Examples of alkynyl groups include prop-2-alkynyl, but-2-alkynyl, but-3-alkynyl, pent-2-alkynyl, 3-methylpent-4-alkynyl, hex-2-alkynyl, hex-5-alkynyl, etc. In the embodiments, the alkynyl group contains a carbon-carbon triple bond. The alkynyl group may be unsubstituted or substituted with one or more substituents as described herein. For example, the alkynyl group may be substituted with one or more of the following (e.g., 1, 2, 3, 4, 5, or 6 independently selected substituents): halogen, -COR", -CO2H, -CO2R", -CN, -OH, -OR", -OCOR", -OCO2R", -NH2, -NHR", -N(R)2, -SR, or -SO2R", wherein each instance of R is independently C1-C2. 20 Aliphatic (e.g., C1-C) 20 Alkyl, C1-C15 Alkyl, C1-C 10 Alkyl or C1-C3 alkyl). In the examples, R” is independently an unsubstituted alkyl group (e.g., unsubstituted C1-C3 alkyl). 20 Alkyl, C1-C 15 Alkyl, C1-C 10 Alkyl or C1-C3 alkyl). In the examples, R” is independently an unsubstituted C1-C3 alkyl. In the examples, the alkynyl group is unsubstituted. In the examples, the alkynyl group is substituted (e.g., substituted by 1, 2, 3, 4, 5 or 6 substituent groups as described herein).

[0081] As used herein, “alkynyl” also refers to a free radical (“C2-C”) of a straight-chain or branched hydrocarbon group having 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). 50 The alkynyl group ("alkynyl group") is also referred to as "ene-alkynyl". In some embodiments, the alkynyl group has 2 to 40 carbon atoms ("C2-C"). 40 The alkynyl group (“acetylinyl”) is present in some embodiments. In some embodiments, the alkynyl group has 2 to 30 carbon atoms (“C2-C”). 30 The alkynyl group (“acetylinyl”) has 2 to 20 carbon atoms (“C2-C”) in some embodiments. 20 The alkynyl group (“acetylation group”) has 2 to 10 carbon atoms (“C2-C”) in some embodiments. 10The alkynyl group has 2 to 9 carbon atoms (“C2-C9 alkynyl”). In some embodiments, the alkynyl group has 2 to 8 carbon atoms (“C2-C8 alkynyl”). In some embodiments, the alkynyl group has 2 to 7 carbon atoms (“C2-C7 alkynyl”). In some embodiments, the alkynyl group has 2 to 6 carbon atoms (“C2-C6 alkynyl”). In some embodiments, the alkynyl group has 2 to 5 carbon atoms (“C2-C5 alkynyl”). In some embodiments, the alkynyl group has 2 to 4 carbon atoms (“C2-C4 alkynyl”). In some embodiments, the alkynyl group has 2 to 3 carbon atoms (“C2-C3 alkynyl”). In some embodiments, the alkynyl group has 2 carbon atoms (“C2 alkynyl”). Multiple carbon-triple bonds may be located internally (e.g., in a 2-butynyl group) or terminally (e.g., in a 1-butynyl group). Examples of C2-C4 alkynyl groups include, but are not limited to, ethynyl (C2), 1-propynyl (C3), 2-propynyl (C3), 1-butynyl (C4), 2-butynyl (C4), etc. Examples of C2-C6 alkenyl groups include the aforementioned C2-C4 alkynyl groups as well as pentynyl (C5), hexynyl (C6), etc. Further examples of alkynyl groups include heptynyl (C7), octyynyl (C8), etc. Unless otherwise stated, each example of an alkynyl group is independently unsubstituted (“unsubstituted alkynyl”) or substituted with one or more substituents (“substituted alkynyl”). In some embodiments, the alkynyl group is an unsubstituted C2-C... 50 Alkyne group. In some embodiments, the alkynyl group is a substituted C2-C group. 50 Alkyne group.

[0082] Aryl: The term "aryl" used alone or as part of a larger portion of a term such as "araneyl" refers to a monocyclic, bicyclic, or tricyclic carbocyclic ring system having a total of six to fourteen ring members, wherein the ring system has a single connection point to the rest of the molecule, at least one ring in the system is aromatic, and each ring in the system contains four to seven ring members. In embodiments, the aryl group has six ring carbon atoms ("C6 aryl", e.g., phenyl). In some embodiments, the aryl group has ten ring carbon atoms ("C6 aryl"). 10 "Aryl", for example, naphthyl, such as 1-naphthyl and 2-naphthyl). In some embodiments, the aryl group has 14 cyclic carbon atoms ("C"). 14 "Aryl" (e.g., anthracene). "Aryl" also includes ring systems in which an aromatic ring as defined above is fused with one or more carbocyclic or heterocyclic groups, wherein the linking radical or linking point is on the aromatic ring, and in this case, the number of carbon atoms continues to represent the number of carbon atoms in the aromatic ring system. Exemplary aryl groups include phenyl, naphthyl, and anthracene.

[0083] As used herein, “aryl” also refers to a radical (“C6-C”) of a monocyclic or polycyclic (e.g., bicyclic or tricyclic) 4n+2 aromatic ring system (e.g., having 6, 10, or 14π electrons shared in the ring array) having 6 to 14 ring carbon atoms and zero heteroatoms provided in the aromatic ring system. 14 Aryl group (“C6 aryl”). In some embodiments, the aryl group has 6 ring carbon atoms (“C6 aryl”; for example, phenyl). In some embodiments, the aryl group has 10 ring carbon atoms (“C6 aryl”). 10 "Aryl"; for example, naphthyl, such as 1-naphthyl and 2-naphthyl). In some embodiments, the aryl group has 14 ring carbon atoms ("C"). 14 "Aryl"; for example, anthracene. "Aryl" also includes ring systems in which an aromatic ring as defined above is fused with one or more carbocyclic or heterocyclic groups, wherein the linking radical or linker is located on the aromatic ring, and in this case, the number of carbon atoms continues to represent the number of carbon atoms in the aromatic ring system. Unless otherwise stated, each instance of an aryl is independently unsubstituted ("unsubstituted aryl") or substituted with one or more substituents ("substituted aryl"). In some embodiments, the aryl is an unsubstituted C6-C 14 Aryl group. In some embodiments, the aryl group is a substituted C6-C group. 14 Aryl.

[0084] arylene: As used herein, the term "arylene" refers to a divalent aryl group (i.e., having two connection sites with the molecule). Exemplary arylenes contain phenylene (e.g., unsubstituted or substituted phenylene).

[0085] Carbocyclic group: As used herein, "carbocyclic group" or "carbocyclic" refers to a non-aromatic ring system having 3 to 10 ring carbon atoms ("C3-C"). 10 A free radical consisting of a carbocyclic group (“C3-C8 carbocyclic”) and a non-aromatic cyclic hydrocarbon group with zero heteroatoms. In some embodiments, the carbocyclic group has 3 to 8 cyclic carbon atoms (“C3-C7 carbocyclic”). In some embodiments, the carbocyclic group has 3 to 7 cyclic carbon atoms (“C3-C7 carbocyclic”). In some embodiments, the carbocyclic group has 3 to 6 cyclic carbon atoms (“C3-C6 carbocyclic”). In some embodiments, the carbocyclic group has 4 to 6 cyclic carbon atoms (“C4-C6 carbocyclic”). In some embodiments, the carbocyclic group has 5 to 6 cyclic carbon atoms (“C5-C6 carbocyclic”). In some embodiments, the carbocyclic group has 5 to 10 cyclic carbon atoms (“C5-C6 carbocyclic”). 10The C3-C6 carbocyclic group includes, but is not limited to, cyclopropyl (C3), cyclopropenyl (C3), cyclobutyl (C4), cyclobutenyl (C4), cyclopentyl (C5), cyclopentenyl (C5), cyclohexyl (C6), cyclohexenyl (C6), cyclohexadienyl (C6), etc. The C3-C8 carbocyclic group includes, but is not limited to, the aforementioned C3-C6 carbocyclic groups, as well as cycloheptyl (C7), cycloheptenyl (C7), cycloheptadienyl (C7), cycloheptanetrienyl (C7), cyclooctyl (C8), cyclooctenyl (C8), bicyclo[2.2.1]heptyl (C7), bicyclo[2.2.2]octyl (C8), etc. The C3-C6 carbocyclic group includes, but is not limited to, cyclopropyl (C3), cyclopropenyl (C3), cyclobutyl (C4), cyclobutenyl (C4), cyclopentenyl (C5), cyclopentenyl (C5), cyclohexyl (C6), cyclohexenyl (C6), bicyclo[2.2.2]octyl (C8), etc. 10 Carbocyclic groups include, but are not limited to, the aforementioned C3-C8 carbocyclic groups, as well as cyclononyl (C9), cyclononenyl (C9), and cyclodecyl (C9). 10 ), cyclodecenyl (C 10 ), octahydro-1H-indenyl (C9), decahydronaphthyl (C9) 10 ), spiro[4.5]decyl(C 10 As illustrated in the foregoing examples, in some embodiments, the carbocyclic group is monocyclic (“monocyclic carbocyclic”) or polycyclic (e.g., containing fused, bridged, or spirocyclic systems, such as bicyclic systems (“bicyclic carbocyclic”) or tricyclic systems (“tricyclic carbocyclic”)), and may be saturated or may contain one or more carbon-carbon double or triple bonds. “Carbocyclic” also includes ring systems in which the carbocyclic ring as defined above is fused with one or more aryl or heteroaryl groups, wherein the attachment point is on the carbocyclic ring, and in these cases, the number of carbons always represents the number of carbons in the carbocyclic system. Unless otherwise stated, each instance of a carbocyclic group is independently unsubstituted (“unsubstituted carbocyclic”) or substituted with one or more substituents (“substituted carbocyclic”). In some embodiments, the carbocyclic group is unsubstituted C3-C 10 Carbocyclic group. In some embodiments, the carbocyclic group is a substituted C3-C group. 10 Carbon cyclic group.

[0086] In some embodiments, "carbocyclic" or "carbocyclic" refers to "cycloalkyl", that is, a monocyclic saturated carbocyclic group ("C3-C4") having 3 to 10 ring carbon atoms. 10 The cycloalkyl group has 3 to 8 cyclic carbon atoms (“C3-C8 cycloalkyl”). In some embodiments, the cycloalkyl group has 3 to 6 cyclic carbon atoms (“C3-C6 cycloalkyl”). In some embodiments, the cycloalkyl group has 4 to 6 cyclic carbon atoms (“C4-C6 cycloalkyl”). In some embodiments, the cycloalkyl group has 5 to 6 cyclic carbon atoms (“C5-C6 cycloalkyl”). In some embodiments, the cycloalkyl group has 5 to 10 cyclic carbon atoms (“C5-C6 cycloalkyl”). 10Examples of C5-C6 cycloalkyl groups include cyclopentyl (C5) and cyclohexyl (C5). Examples of C3-C6 cycloalkyl groups include the aforementioned C5-C6 cycloalkyl groups, 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 stated, each example of a cycloalkyl group is independently unsubstituted (“unsubstituted cycloalkyl”) or substituted with one or more substituents (“substituted cycloalkyl”). In some embodiments, the cycloalkyl group is an unsubstituted C3-C6 cycloalkyl group. 10 Cycloalkyl. In some embodiments, the cycloalkyl group is a substituted C3-C. 10 Cycloalkyl.

[0087] Halogen: As used in this article, the term “halogen” refers to fluorine, chlorine, bromine or iodine.

[0088] Heteroalkyl: The term "heteroalkyl" refers to a branched or unbranched alkyl, alkenyl, or alkynyl group having one to fourteen carbon atoms in addition to one, two, three, or four heteroatoms independently selected from the group consisting of N, O, S, and P. Heteroalkyl groups include tertiary amines, secondary amines, ethers, thioethers, amides, thioamides, carbamates, thiocarbamates, hydrazones, imines, phosphate diesters, aminophosphates, sulfonamides, and disulfides. Heteroalkyl groups may optionally include monocyclic, bicyclic, or tricyclic rings, wherein each ring ideally has three to six members. Examples of heteroalkyl groups include polyethers such as methoxymethyl and ethoxyethyl.

[0089] Heteroalkyl: As used herein, the term “heteroalkyl” refers to the divalent form of a heteroalkyl group as described herein.

[0090] Heteroaryl: As used herein, the term “heteroaryl” is a fully unsaturated heteroatom-containing ring in which at least one ring atom is a heteroatom, such as, but not limited to, nitrogen and oxygen.

[0091] As used herein, “heteroaryl” also refers to a radical (e.g., having 6, 10, or 14π electrons shared in the ring array) of a 5- to 14-membered monocyclic or polycyclic (e.g., bicyclic or tricyclic) 4n+2 aromatic ring system having a cyclic carbon atom provided in the aromatic ring system and one or more (e.g., 1, 2, 3, or 4) cyclic heteroatoms, wherein each heteroatom is independently selected from oxygen, sulfur, nitrogen, boron, silicon, and phosphorus (“5- to 14-membered heteroaryl”). In heteroaryls containing one or more nitrogen atoms, the attachment point may be a carbon or nitrogen atom, where the valence allows. A heteroaryl polycyclic system may contain one or more heteroatoms in one or two rings. “Hyperaryl” includes a ring system in which a heteroaryl ring as defined above is fused with one or more carbocyclic or heterocyclic groups, wherein the attachment point is on the heteroaryl ring, and in such cases, the number of ring members always indicates the number of ring members in the heteroaryl ring system. "Heteroaryl" also includes ring systems in which a heteroaryl ring, as defined above, is fused with one or more aryl groups, wherein the attachment point is on the aryl or heteroaryl ring, and in such cases, the number of ring members indicates the number of ring members in the fused polycyclic (aryl / heteroaryl) ring system. One ring in a polycyclic heteroaryl ring does not contain a heteroatom (e.g., indolyl, quinolinyl, carbazolyl, etc.), and the attachment point can be on either ring, i.e., a ring with a heteroatom (e.g., 2-indolyl) or a ring without a heteroatom (e.g., 5-indolyl).

[0092] In some embodiments, a heteroaryl group is a 5- to 10-membered aromatic ring system having a cyclic carbon atom provided in the aromatic ring system and one or more (e.g., 1, 2, 3, 4) cyclic heteroatoms, wherein each heteroatom is independently selected from oxygen, sulfur, nitrogen, boron, silicon, and phosphorus (“5- to 10-membered heteroaryl”). In some embodiments, a heteroaryl group is a 5- to 8-membered aromatic ring system having a cyclic carbon atom provided in the aromatic ring system and one or more (e.g., 1, 2, 3, 4) cyclic heteroatoms, wherein each heteroatom is independently selected from oxygen, sulfur, nitrogen, boron, silicon, and phosphorus (“5- to 8-membered heteroaryl”). In some embodiments, a heteroaryl group is a 5- to 6-membered aromatic ring system having a cyclic carbon atom provided in the aromatic ring system and one or more (e.g., 1, 2, 3, 4) cyclic heteroatoms, wherein each heteroatom is independently selected from oxygen, sulfur, nitrogen, boron, silicon, and phosphorus (“5- to 6-membered heteroaryl”). In some embodiments, the 5- to 6-membered heteroaryl group has one or more (e.g., one, two, or three) cyclic heteroatoms selected from oxygen, sulfur, nitrogen, boron, silicon, and phosphorus. In some embodiments, the 5- to 6-membered heteroaryl group has one or two cyclic heteroatoms selected from oxygen, sulfur, nitrogen, boron, silicon, and phosphorus. In some embodiments, the 5- to 6-membered heteroaryl group has one cyclic heteroatom selected from oxygen, sulfur, nitrogen, boron, silicon, and phosphorus. Unless otherwise stated, each instance of a heteroaryl group is independently unsubstituted (“unsubstituted heteroaryl”) or substituted with one or more substituents (“substituted heteroaryl”). In some embodiments, the heteroaryl group is an unsubstituted 5- to 14-membered heteroaryl group. In some embodiments, the heteroaryl group is a substituted 5- to 14-membered heteroaryl group.

[0093] Exemplary 5-membered heteroaryl groups containing one heteroatom include, but are not limited to, pyrroleyl, furanyl, and thiopheneyl. Exemplary 5-membered heteroaryl groups containing two heteroatoms include, but are not limited to, imidazolyl, pyrazolyl, oxazolyl, isoxazolyl, thiazolyl, and isothiazolyl. Exemplary 5-membered heteroaryl groups containing three heteroatoms include, but are not limited to, triazolyl, oxadiazolyl, and thiadiazolyl. Exemplary 5-membered heteroaryl groups containing four heteroatoms include, but are not limited to, tetrazolyl. Exemplary 6-membered heteroaryl groups containing one heteroatom include, but are not limited to, pyridinyl. Exemplary 6-membered heteroaryl groups containing two heteroatoms include, but are not limited to, pyridazinyl, pyrimidinyl, and pyrazinyl. Exemplary 6-membered heteroaryl groups containing three or four heteroatoms include, but are not limited to, triazinyl and tetraazinyl. Exemplary 7-membered heteroaryl groups containing one heteroatom include, but are not limited to, azirmonyl, oxadimonyl, and thiopheneyl. Exemplary 5,6-bicyclic heteroaryl groups include, but are not limited to, indolyl, isoindolyl, indazole, benzotriazolyl, benzothiophenyl, isobenzothiophenyl, benzofuranyl, benzoisofuranyl, benzoimidazolyl, benzoxazolyl, benzoisoxazolyl, benzoxadiazole, benzothiazolyl, benzoisothiazolyl, benzothiadiazolyl, indazinyl, and purinyl. Exemplary 6,6-bicyclic heteroaryl groups include, but are not limited to, naphridinyl, piperidinyl, quinolinyl, isoquinolinyl, quinolinyl, quinoxalinyl, phthalazinyl, and quinazolinyl. Exemplary tricyclic heteroaryl groups include, but are not limited to, phenanthridinel, dibenzofuranyl, carbazoleyl, acridinel, phenothiazinyl, phenotoxazinyl, and phenotazinyl.

[0094] As used herein, a “heterocyclic group” or “heterocycle” refers to a radical of a 3- to 14-membered nonaromatic ring system having a ring carbon atom and one or more (e.g., one, two, three, or four) ring heteroatoms, wherein each heteroatom is independently selected from oxygen, sulfur, nitrogen, boron, silicon, and phosphorus (“3- to 14-membered heterocyclic groups”). In heterocyclic groups containing one or more nitrogen atoms, the attachment point may be a carbon atom or a nitrogen atom, where the valence allows. Heterocyclic groups may be monocyclic (“monocyclic heterocyclic groups”) or polycyclic (e.g., fused, bridged, or spirocyclic systems, such as bicyclic systems (“bicyclic heterocyclic groups”) or tricyclic systems (“tricyclic heterocyclic groups”)) and may be saturated or may contain one or more carbon-carbon double or triple bonds. Heterocyclic polycyclic systems may contain one or more heteroatoms in one or two rings. "Heterocyclic group" also includes ring systems, wherein a heterocyclic ring as defined above is fused with one or more carbocyclic groups, wherein the attachment point is on the carbocyclic group or the heterocyclic ring, or the ring system; wherein a heterocyclic ring as defined above is fused with one or more aryl or heteroaryl groups, wherein the attachment point is on the heterocyclic ring; and in such cases, the number of ring members always represents the number of ring members in the heterocyclic ring system. Unless otherwise stated, each instance of a heterocyclic group is independently unsubstituted ("unsubstituted heterocyclic group") or substituted with one or more substituents ("substituted heterocyclic group"). In some embodiments, the heterocyclic group is an unsubstituted 3- to 14-membered heterocyclic group. In some embodiments, the heterocyclic group is a substituted 3- to 14-membered heterocyclic group.

[0095] In some embodiments, the heterocyclic group is a 5-10 membered non-aromatic ring system having a cyclic carbon atom and one or more (e.g., 1, 2, 3, or 4) cyclic heteroatoms, wherein each heteroatom is independently selected from oxygen, sulfur, nitrogen, boron, silicon, and phosphorus (“5-10 membered heterocyclic group”). In some embodiments, the heterocyclic group is a 5-8 membered non-aromatic ring system having a cyclic carbon atom and one or more (e.g., 1, 2, 3, or 4) cyclic heteroatoms, wherein each heteroatom is independently selected from oxygen, sulfur, nitrogen, boron, silicon, and phosphorus (“5-8 membered heterocyclic group”). In some embodiments, the heterocyclic group is a 5-6 membered non-aromatic ring system having a cyclic carbon atom and one or more (e.g., 1, 2, 3, or 4) cyclic heteroatoms, wherein each heteroatom is independently selected from oxygen, sulfur, nitrogen, boron, silicon, and phosphorus (“5-6 membered heterocyclic group”). In some embodiments, the 5- to 6-membered heterocyclic group has one or more (e.g., one, two, or three) cyclic heteroatoms selected from oxygen, sulfur, nitrogen, boron, silicon, and phosphorus. In some embodiments, the 5- to 6-membered heterocyclic group has one or two cyclic heteroatoms selected from oxygen, sulfur, nitrogen, boron, silicon, and phosphorus. In some embodiments, the 5- to 6-membered heterocyclic group has one cyclic heteroatom selected from oxygen, sulfur, nitrogen, boron, silicon, and phosphorus.

[0096] Exemplary 3-membered heterocyclic groups containing one heteroatom include, but are not limited to, aziridinyl, ethylene oxide, and thioenyl. Exemplary 4-membered heterocyclic groups containing one heteroatom include, but are not limited to, azirrobutyl, oxetyl, and thiobutyl. Exemplary 5-membered heterocyclic groups containing one heteroatom include, but are not limited to, tetrahydrofuranyl, dihydrofuranyl, tetrahydrothiophenyl, dihydrothiophenyl, pyrrolyl, dihydropyrrolyl, and pyrrolyl-2,5-diketone. Exemplary 5-membered heterocyclic groups containing two heteroatoms include, but are not limited to, dioxopranyl, oxothiophene, and dithiophene. Exemplary 5-membered heterocyclic groups containing three heteroatoms include, but are not limited to, triazolinyl, oxadiazolinyl, and thiadiazolinyl. Exemplary 6-membered heterocyclic groups containing one heteroatom include, but are not limited to, piperidinyl, tetrahydropyranyl, dihydropyridinyl, and thiophene. Exemplary 6-membered heterocyclic groups containing two heteroatoms include, but are not limited to, piperazinyl, morpholinyl, dithiohexane, and dioxanehexane. Exemplary 6-membered heterocyclic groups containing two heteroatoms include, but are not limited to, triazineyl. Exemplary 7-membered heterocyclic groups containing one heteroatom include, but are not limited to, azirheptanyl, oxetaneheptyl, and thioheptanyl. Exemplary 8-membered heterocyclic groups containing one heteroatom include, but are not limited to, azirheptanyl, oxetaneheptyl, and thioheptanyl. Exemplary bicyclic heterocyclic groups include, but are not limited to, indole, isoindole, dihydrobenzofuranyl, dihydrobenzothiophenyl, tetrahydrobenzothiophenyl, tetrahydrobenzofuranyl, tetrahydroindole, tetrahydroquinolinyl, tetrahydroisoquinolinyl, decahydroquinolinyl, decahydroisoquinolinyl, octahydrochromenyl, octahydroisochromenyl, decahydronaphthyridyl, decahydro-1,8-naphthyridyl, decahydropyrrolo[3,2-b]pyrrole, indole, phthalimide, naphthimide, benzodihydropyranyl, benzothiophene, 1H-benzo[e][1,4]diazapyridyl, 1,4,5,7-tetrahydropyranolo[3,4-b]pyrrole. 5,6-Dihydro-4H-furano[3,2-b]pyrrolithyl, 6,7-Dihydro-5H-furano[3,2-b]pyrrolithyl, 5,7-Dihydro-4H-thieno[2,3-c]pyrrolithyl, 2,3-Dihydro-1H-pyrroli[2,3-b]pyridyl, 2,3-Dihydrofurano[2,3-b]pyridyl, 4,5,6,7-Tetrahydro-1H-pyrroli[2,3-b]pyridyl, 4,5,6,7-Tetrahydrofurano[3,2-c]pyridyl, 4,5,6,7-Tetrahydrothieno[3,2-b]pyridyl, 1,2,3,4-Tetrahydro-1,6-naphthidyl, etc.

[0097] Heterocyclic alkyl: As used herein, the term "heterocyclic alkyl" is a non-aromatic ring in which at least one atom is a heteroatom, such as, but not limited to, nitrogen, oxygen, sulfur, or phosphorus, and the remaining atoms are carbon. Heterocyclic alkyl can be substituted or unsubstituted.

[0098] As understood above, alkyl, alkenyl, alkynyl, acyl, carbocyclic, heterocyclic, aryl, and heteroaryl groups as defined herein are optionally substituted in some embodiments. Optional substitution means that the group 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” carbocyclic, “substituted” or “unsubstituted” heterocyclic, “substituted” or “unsubstituted” aryl, or “substituted” or “unsubstituted” heteroaryl). Generally, the term "substituted" refers to a group in which at least one hydrogen atom is substituted by a permissible substituent, such as a substituent that, upon substitution, produces a stable compound, for example, a compound that does not spontaneously undergo transformation by rearrangement, cyclization, elimination, or other reactions. Unless otherwise stated, a "substituted" group has a substituent at one or more substituted 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 encompass substitution with all permissible substituents of an organic compound, resulting in the formation of a stable compound by any of the substituents described herein. The invention contemplates any and all of these combinations to obtain a stable compound. For the purposes of this invention, a heteroatom (such as nitrogen) may have a hydrogen substituent and / or satisfy the valence of the heteroatom and such that a suitable substituent as described herein forms a stable moiety.

[0099] 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(Rbb2), -NRbbC(=O)Raa, -NR bbCO2Raa, -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)(N Rbb)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 carbocyclic, 3-14 heterocyclic, C6-C14 aryl and 5-14 heteroaryl, wherein each alkyl, alkenyl, alkynyl, carbocyclic, heterocyclic, aryl and heteroaryl is independently substituted by 0, 1, 2, 3, 4 or 5 Rdd groups;

[0100] Alternatively, the two twin hydrogens on the carbon atom can be replaced by the groups =O, =S, =NN(Rbb)2, =NNRbbC(=O)Raa, =NNRbbC(=O)ORaa, =NNRbbS(=O)2Raa, =NRbb, or =NORcc;

[0101] Each instance of Raa is independently selected from C1-C50 alkyl, C2-C50 alkenyl, C2-C50 alkynyl, C3-C10 carbocyclic, 3-14-membered heterocyclic, C6-C14 aryl, and 5-14-membered heteroaryl, or two Raa groups are linked to form a 3-14-membered heterocyclic or 5-14-membered heteroaryl ring, wherein each alkyl, alkenyl, alkynyl, carbocyclic, heterocyclic, aryl, and heteroaryl group is independently substituted by 0, 1, 2, 3, 4, or 5 Rdd groups;

[0102] Each instance of Rbb is 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 carbocyclic, 3-14-membered heterocyclic, C6-C14 aryl and 5-14-membered heteroaryl, or two Rbb groups and the heteroatom attached thereto form a 3-14-membered heterocyclic or 5-14-membered heteroaryl ring, wherein each alkyl, alkenyl, alkynyl, carbocyclic, heterocyclic, aryl and heteroaryl group is independently substituted by 0, 1, 2, 3, 4 or 5 Rdd groups;

[0103] Each instance of Rcc is independently selected from hydrogen, C1-C50 alkyl, C2-C50 alkenyl, C2-C50 alkynyl, C3-C10 carbocyclic, 3-14-membered heterocyclic, C6-C14 aryl, and 5-14-membered heteroaryl, or two Rcc groups together with the heteroatom attached to them form a 3-14-membered heterocyclic or 5-14-membered heteroaryl ring, wherein each alkyl, alkenyl, alkynyl, carbocyclic, heterocyclic, aryl, and heteroaryl group is independently substituted by 0, 1, 2, 3, 4, or 5 Rdd groups;

[0104] Each instance of Rdd is independently selected from halogens, -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)(Re e)2、-OP(=O)(ORee)2、C1-C50 alkyl、C2-C50 alkenyl、C2-C50 alkynyl、C3-C10 carbocyclic、3-10 heterocyclic、C6-C10 aryl、5-10 heteroaryl、wherein each alkyl、alkenyl、alkynyl、carbocyclic、heterocyclic、aryl、heteroaryl is independently substituted by 0, 1, 2, 3, 4 or 5 Rgg groups, or may be connected with two twin Rdd substituents to form =O or =S;

[0105] Each instance of Ree is independently selected from C1-C50 alkyl, C2-C50 alkenyl, C2-C50 alkynyl, C3-C10 carbocyclic, C6-C10 aryl, 3-10 heterocyclic and 3-10 heteroaryl groups, wherein each alkyl, alkenyl, alkynyl, carbocyclic, heterocyclic, aryl and heteroaryl group is independently substituted by 0, 1, 2, 3, 4 or 5 Rgg groups;

[0106] Each instance of Rff is independently selected from hydrogen, C1-C50 alkyl, C2-C50 alkenyl, C2-C50 alkynyl, C3-C10 carbocyclic, 3-10 heterocyclic, C6-C10 aryl, and 5-10 heteroaryl, or two Rff groups together with the heteroatom they are attached to form a 3-14 heterocyclic or 5-14 heteroaryl ring, wherein each alkyl, alkenyl, alkynyl, carbocyclic, heterocyclic, aryl, and heteroaryl group is independently substituted by 0, 1, 2, 3, 4, or 5 Rgg groups; and

[0107] Each instance of Rgg is independently a halogen, -CN, -NO2, -N3, -SO2H, -SO3H, -OH, -OC1-C50 alkyl, -ON(C1-C50 alkyl)2, -N(C1-C50 alkyl)2, -N(C1-C50 alkyl)3+X-, -NH(C1-C50 alkyl)2+X-, -NH2(C1-C50 alkyl)+X-, -NH3+X-, -N(OC1-C50 alkyl)(C1-C50 alkyl), -N(OH)(C1- C50 alkyl), -NH(OH), -SH, -SC1-C50 alkyl, -SS(C1-C50 alkyl), -C(=O)(C1-C50 alkyl), -CO2H, -CO2(C1-C50 alkyl), -OC(=O)(C1-C50 alkyl), -OCO2(C1-C50 alkyl), -C(=O)NH2, -C(=O)N(C1-C50 alkyl)2, -OC(=O)NH(C1-C50 alkyl), -NHC(=O)(C1-C50 alkyl) -0 alkyl), -N(C1-C50 alkyl)C(=O)(C1-C50 alkyl), -NHCO2(C1-C50 alkyl), -NHC(=O)N(C1-C50 alkyl)2, -NHC(=O)NH(C1-C50 alkyl), -NHC(=O)NH2, -C(=NH)O(C1-C50 alkyl), -OC(=NH)(C1-C50 alkyl), -OC(=NH)OC1-C50 alkyl, -C(=NH)N(C1-C50 alkyl)2 -C(=NH)NH (C1-C50 alkyl), -C(=NH)NH2, -OC(=NH)N (C1-C50 alkyl)2, -OC(NH)NH (C1-C50 alkyl), -OC(NH)NH2, -NHC(NH)N (C1-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 (C1-C6 alkyl), -SO (C1-C6 alkyl), -Si (C1-C50 alkyl)3, -OSi (C1-C6 alkyl)3, -C(=S)N (C1-C50 alkyl)2, C(=S)NH (C1-C50 alkyl), C(=S)NH2, -C(=O)S (C1-C6 alkyl), -C(=S)S (C1-C6 alkyl), -SC(=S)S (C1-C50 alkyl) -C6 alkyl), -P(=O)2(C1-C50 alkyl), -P(=O)(C1-C50 alkyl)2, -OP(=O)(C1-C50 alkyl)2, -OP(=O)(OC1-C50 alkyl)2, C1-C50 alkyl, C2-C50 alkenyl, C2-C50 alkynyl, C3-C10 carbocyclic, C6-C10 aryl, 3-10 heterocyclic, 5-10 heteroaryl; or two twin Rgg substituents can be linked to form =O or =S; where X- is a counterion.

[0108] As used herein, the term “halogen” or “halogen” refers to fluorine (fluorinated, -F), chlorine (chlorinated, -Cl), bromine (brominated, -Br), or iodine (iodinated, -I).

[0109] As used herein, a “counterion” is a negatively charged group that associates with a positively charged quaternary ammonium 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-camphorsulfonate, naphthalene-2-sulfonate, naphthalene-1-sulfonate-5-sulfonate, acetyl-1-sulfonate-2-sulfonate, etc.) and carboxylate ions (e.g., acetate, acetate, propionate, benzoate, glycerate, lactate, tartrate, glycolate, etc.).

[0110] Where valence permits, the nitrogen atom can be substituted or unsubstituted, and may 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, and -P(=O)(Raa). 2. -P(=O)2N(Rcc)2, -P(=O)(NRcc)2, C1-C50 alkyl, C2-C50 alkenyl, C2-C50 alkynyl, C3-C10 carbocyclic, 3-14 heterocyclic, C6-C14 aryl and 5-14 heteroaryl, or two Rcc groups together with the N atom to which they are attached to form a 3-14 heterocyclic or 5-14 heteroaryl ring, wherein each alkyl, alkenyl, alkynyl, carbocyclic, heterocyclic, aryl and heteroaryl group is independently substituted by 0, 1, 2, 3, 4 or 5 Rdd groups, and wherein Raa, Rbb, Rcc and Rdd are as defined above.

[0111] In some embodiments, the substituents present on the nitrogen atom are nitrogen protecting groups (also known as amino protecting groups). Nitrogen protecting groups are those well known in the art and include those described in detail in Protecting Groups in Organic Synthesis, TW Greene and PGM Watts, 3rd Edition, John Wiley & Sons, 1999, which are incorporated herein by reference.

[0112] 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-phenylpropionamide, pyridine amide, 3-pyridylformamide, N-benzoylphenylalanyl derivatives, benzamide, p-phenylbenzamide, o-nitrophenylacetamide, o-nitrophenoxyacetamide, acetylacetamide, (N'-dithiobenzyloxyacetamide)acetamide, 3-(p-hydroxyphenyl)propionamide, 3-(o-nitrophenyl)propionamide, 2-methyl-2-(o-nitrophenoxy)propionamide, 2-methyl-2-(o-phenylphenoxy)propionamide, 4-chlorobutyramide, 3-methyl-3-nitrobutyramide, o-nitrocinnamamide, N-acetylated methionine derivatives, o-nitrobenzamide, and o-(benzoyl)benzamide.

[0113] Nitrogen-protecting groups such as urethane groups (e.g., -C(=O)ORaa) include, but are not limited to: methyl carbamate, ethyl carbamate, 9-fluorenylmethyl carbamate (Fmoc), 9-(2-sulfonyl)fluorenylmethyl carbamate, 9-(2,7-dibromo)fluoroenylmethyl carbamate, 2,7-di-tert-butyl-[9-(10,10-dioxo-10,10,10,10-tetrahydrothioxanthanna)]methyl carbamate (DBD-Tmoc), 4-methoxybenzoyl carbamate (Phenoc), 2,2,2-trichloroethyl carbamate (Troc), 2-trimethylsilylethyl carbamate, etc. Ester (Teoc), 2-Phenylacetic Ethyl Carbamate (hZ), 1-(1-adamantyl)-1-methylethylcarbamate (Adpoc), 1,1-Dimethyl-2-ethylcarbamate, 1,1-Dimethyl-2,2-dibromoethylcarbamate (DB-t-BOC), 1,1-Dimethyl-2,2,2-trichloroethylcarbamate (TCBOC), ethyl 1-methyl-1-(4-biphenyl)carbamate (Bpoc), 1-(3,5-di-tert-butylphenyl)-1-methylethylcarbamate (t-Bumeoc), ethyl 2-(2'- and 4'-pyridyl)carbamate (Pyoc) ), 2-(N,N-dicyclohexylformamido)carbamate, tert-butyl carbamate (BOC), 1-adamantyl carbamate (Adoc), vinyl carbamate (Voc), allyl carbamate (Alloc), 1-isopropylallyl carbamate (Ipaoc), cinnamyl carbamate (Coc), 4-nitrocinnamyl carbamate (Noc), 8-quinolinyl carbamate, N-hydroxypiperidinyl carbamate, alkyl dithiocarbamate, benzyl carbamate (Cbz), p-methoxybenzyl carbamate (Moz), p-nitrobenzyl carbamate, p-bromobenzyl carbamate, p- Chlorobenzyl carbamate, 2,4-dichlorobenzyl carbamate, 4-methylsulfinylbenzyl carbamate (Msz), 9-anthraylmethyl carbamate, diphenylmethyl carbamate, 2-methylthioethyl carbamate, 2-methylsulfonylethyl carbamate, ethyl 2-(p-toluenesulfonyl)carbamate, [2-(1,3-dithiaalkyl)]methyl carbamate (Dmoc), 4-methylthiophenyl carbamate (Mtpc), 2,4-dimethylthiophenyl carbamate (Bmpc), 2-phosphonoethyl carbamate (Peoc), 2-triphenylphosphonoisopropyl carbamate (Ppoc), 1,1-Dimethyl-2-cyanoethyl carbamate, m-chloro-p-acyloxybenzyl carbamate, p-(dihydroxyformyl)benzyl carbamate, 5-benzisoxazolyl methyl carbamate, 2-(trifluoromethyl)-6-bromomethyl carbamate (Tcroc), m-nitrophenyl carbamate, 3,5-dimethoxybenzyl carbamate, o-carbamate nitrobenzyl ester, 3,4-dimethoxy-6-nitrobenzyl carbamate, phenyl (o-nitro) 1,1-dimethyl-3-(N,N-dimethylformamido)carbamate, tert-amylcarbamate, S-benzylthiocarbamate, p-cyanobenzylcarbamate, cyclobutylcarbamate, cyclohexylcarbamate, cyclopentylcarbamate, cyclopropylmethylcarbamate, p-decoxybenzylcarbamate, 2,2-dimethoxyylvinylcarbamate, o-(N,N-dimethylformamido)carbamate, 1,1-dimethyl-3-(N,N-dimethylformamido)propylcarbamate 1,1-Dimethylpropynylcarbamate, 2-pyridylmethylcarbamate, 2-furanylmethylcarbamate, 2-iodoethylcarbamate, isobutylcarbamate, isobutylcarbamate, isonicotinamide carbamate, p-(p'-methoxyphenylazo)benzylcarbamate, 1-methylcyclobutylcarbamate, 1-methylcyclohexylcarbamate, 1-1-methyl-1-cyclopropylmethylcarbamate 1-Methyl-1-(3,5-dimethoxyphenyl)ethylcarbamate, 1-methyl-1-(p-phenylazophenyl)ethylcarbamate, 1-methyl-1-phenylethylcarbamate, ethyl 1-methyl-1-(4-pyridyl)carbamate, phenyl carbamate, benzyl p-(phenylazo)carbamate, 2,4,6-tri-tert-butylphenyl carbohydrate, 4-(trimethylammonium)benzylcarbamate, and 2,4,6-trimethylbenzylcarbamate.

[0114] 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), and 4-methoxybenzenesulfonamide (Mbs). ), 2,4,6-trimethylbenzenesulfonamide (Mts), 2,6-dimethoxy-4-methylbenzenesulfonamide (iMds), 2,2,5,7,8-pentamethylbenzodihydropyran-6-sulfonamide (Pmc), methanesulfonamide (Ms), β-trimethylsilylsulfonamide (SES), 9-anthracitesulfonamide, 4-(4',8'-dimethoxynaphthylmethyl)benzenesulfonamide (DNMBS), benzylsulfonamide, trifluoromethylsulfonamide and benzoylsulfonamide.

[0115] Other nitrogen-protecting groups include, but are not limited to: phenthiazinyl-(10)-acyl derivatives, N'-p-toluenesulfonylaminoacyl derivatives, N'-phenylaminothioacyl derivatives, N-benzoylphenylalanyl derivatives, N-acetylmethionine derivatives, 4,5-diphenyl-3-oxazoline-2-one, N-phthalimide, N-dithiopheneimide (Dts), N-2,3-diphenylmaleimide, N-2,5-dimethylpyrrole, N-1,1,4,4-tetramethyldisiloxylcyclopentane adduct (STABASE), 5-substituted 1,3-dimethyl-1,3,5-triazacyclohexane-2-one, 5 -substituted 1,3-dibenzyl-1,3,5-triazacyclohexane-2-one, 1-substituted 3,5-dinitro-4-pyridinone, N-methylamine, N-allylamine, N-[2-(trimethylsilyl)ethoxy]methylamine (SEM), N-3-acetoxypropylamine, N-(1-isopropyl-4-nitro-2-oxo-3-pyrrololin-3-yl)amine, quaternary ammonium salts, N-benzylamine, N-di(4-methoxyphenyl)methylamine, N-5-dibenzofuranamine, N-triphenylmethylamine (Tr), N-[(4-methoxyphenyl)diphenylmethyl]amine (MMTr), N-9-phenylfluorofluoreneamine (PhF), N-2,7-dichloro- 9-fluorenylmethyleneamine, N-ferrocenemethylamino (Fcm), N-2-pyridylamino N'-oxide, N-1,1-dimethylthiomethyleneamine, N-benzylamine, Np-methoxybenzylamine, N-diphenylmethyleneamine, N-[(2-pyridyl)trimethyl]methyleneamine, N-(N',N'-dimethylaminomethylene)amine, N,N'-isopropylidenediamine, Np-nitrobenzylamine, N-salicylamine, N-5-chlorosulfinamide, N-(5-chloro-2-hydroxyphenyl)phenylmethyleneamine, N-cyclohexylamine, N-(5,5-dimethyl-3-oxo-1-cyclohexenyl)amine, N-borane derivatives, N-diphenylboronic acid derivatives, N-[phenyl(pentaacylchromium- or tungsten)acyl]amine, N-copper chelate, N-zinc chelate, N-nitroamine, N-nitrosoamine, amine N-oxide, diphenylphosphamide (Dpp), dimethylthiophosphamide (Mpt), diphenylthiophosphamide (Ppt), dialkylaminophosphate, dibenzylaminophosphate, diphenylaminophosphate, benzenesulfonamide, o-nitrobenzenesulfonamide (Nps), 2,4-dinitrobenzenesulfonamide, pentachlorobenzenesulfonamide, 2-nitro-4-methoxybenzenesulfonamide, triphenylmethylsulfinamide, and 3-nitropyridinesulfinamide (Npys).

[0116] In some embodiments, the substituent present on the oxygen atom is an oxygen protecting group (also known 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, TW Greene and PGM Uts, 3rd Edition, John Wiley & Sons, 1999, which are incorporated herein by reference.

[0117] Exemplary oxygen protecting groups include, but are not limited to: methyl, methoxymethyl (MOM), methylthiomethyl (MTM), tert-butylthiomethyl, (phenyldimethylsilyl)methoxymethyl (SMOM), benzyloxymethyl (BOM), p-methoxybenzyloxymethyl (PMBM), (4-methoxyphenoxy)methyl (p-AOM), guaiacol methyl (GUM), tert-butoxymethyl, 4-pentenoxymethyl (POM), siloxymethyl, 2-methoxyethoxymethyl (MEM), 2,2,2-trichloroethoxymethyl, bis(2-chloroethoxy)methyl, 2-(trimethylsilyl)ethoxymethyl (SEMOR), tetrahydropyridine THP, 3-bromotetrahydropyranyl, tetrahydrothiaranyl, 1-methoxycyclohexyl, 4-methoxytetrahydropyranyl (MTHP), 4-methoxytetrahydrothiaranyl, 4-methoxytetrahydrothiaranyl S, S-dioxide, 1-[(2-chloro-4-methyl)phenyl]-4-methoxypiperidin-4-yl (CTMP), 1,4-dioxane-2-yl, tetrahydrofuranyl, tetrahydrothiopheneyl, 2,3,3a,4,5,6,7,7α-octahydro-7,8,8-trimethyl-4,7-methylenebenzofuran-2-yl, 1-ethoxyethyl, 1-(2-chloroethoxy)ethyl, 1-methyl-1-methoxyethyl, 1-methyl 1-Benzyloxyethyl, 1-methyl-1-benzyloxy-2-fluoroethyl ester, 2,2,2-trichloroethyl, 2-trimethylsilylethyl, 2-(phenylselenoethyl)ethyl, tert-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-pyridinylmethyl, 4-pyridinylmethyl, 3-methyl-2-pyridinylmethyl N-oxonium, diphenylmethyl, p,p'-dinitrodiphenylmethyl, 5-dibenzocycloheptan, triphenylmethyl, α-naphthalene diphenylmethyl, p-methoxyphenyl diphenylmethyl, di(p-methoxyphenyl)phenylmethyl, tri(p-methoxyphenyl)methyl, 4-(4'-bromobenzoylmethylphenoxy)diphenylmethyl, 4,4',4”-tris(4,5-dichlorophthaliminophenyl)methyl, 4,4',4”-tris(acetylpropionyloxyphenyl)methyl, 4,4',4”-tris(benzyloxy)methyl, 3-(imidazol-1-yl)bis(4',4”-dimethoxyphenyl)methyl, 1,1-bis(4-methoxyphenyl)-1'-pyrenemethyl, 9-anthrayl, 9-(9-phenyl)tonyl, 9-(9-phenyl-10-oxo)anthrayl, 1,3-Benzodithiofuran-2-yl, Benzisothiazolyl S, S-dioxane, Trimethylsilyl (TMS), Triethylsilyl (TES), Triisopropylsilyl (TIPS), Dimethylisopropyl (IPDMS), Diethylisopropyl (DEIPS), Dimethylhexylsilyl, Tert-butyldimethylsilyl (TBDMS), Tert-butyldiphenylsilyl (TBDPS), Tribenzylsilyl, Tri-p-xylylsilyl, Triphenylsilyl, Diphenylmethyl (DPMS), Tert-butylmethoxyphenylsilyl (TBMPS), Formate, Benzoyl, Acetate, Chloroacetic acid, Dichloroacetic acid, Trichloroacetic acid, Trifluoroacetate, Methoxyacetic acid, Triphenylmethoxyacetic acid Phenoxy, p-chlorophenoxy, 3-phenylpropionate, 4-oxovalerate (acetylpropionate), 4,4-(ethylidene)valerate (L-dithioacetal), neovalerate, adamantane, crotonate, methyl 4-crotonate, benzoate, p-phenyl, 2,4,6-trimethylbenzoate (2,4,6-trimethylbenzoate or mesitoate), alkyl methyl carbonate, 9-fluorenyl ester (Fmoc), alkyl ethyl carbonate, alkyl 2,2,2-trichloroethyl carbonate (Troc), 2-(trimethylsilyl)ethyl carbonate (TMSEC), 2-(phenylsulfonyl)ethyl carbonate (Psec), 2-(triphenylphosphino)ethyl carbonate Ethyl carbonate (Peoc), alkyl isobutyl carbonate, alkyl ethylene carbonate, alkyl allyl carbonate, alkyl p-nitrobenzene carbonate, alkyl benzyl carbonate, alkyl p-methoxybenzyl carbonate, alkyl 3,4-dimethoxybenzyl carbonate, alkyl o-nitrobenzyl carbonate, alkyl p-nitrobenzyl carbonate, alkyl S-thiobenzyl carbonate, 4-ethoxy-1-naphthyl carbonate, dithiocarbonate, methyl 2-iodobenzoic acid, 4-azidobutyrate, 4-nitro-4-methylvalerate, o-(dibromomethyl)benzoate, 2-formylbenzenesulfonate, 2-(methylthiomethoxy)ethyl, 4-(methylthiomethoxy)butyrate, 2-(methylthiomethoxymethyl)benzoate, 2,6- Dichloro-4-methylphenoxyacetic acid ester, 2,6-dichloro-4-(1,1,3,3-tetramethylbutyl)phenoxyacetic acid ester, 2,4-bis(1,1-dimethylpropyl)phenoxyacetic acid ester, chlorodiphenylacetic acid ester, isobutyrate, monomalate, (E)-2-methyl-2-butenoate, o-(methoxy)benzoate, α-naphthalene, nitrates, alkyl N,N,N',N'-tetramethylphosphoric acid diamine, alkyl N-phenylcarbamate, borates, dimethylphosphine thiocyanate, alkyl 2,4-dinitrophenyl sulfinates, sulfates, methanesulfonate (or mesylate), benzyl sulfonate, and toluenesulfonate (Ts).

[0118] In some embodiments, the substituent present on the sulfur atom is a sulfur protecting group (also known 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, TW Greene and PGM Uts, 3rd Edition, John Willie & Sons, 1999, which are incorporated herein by reference.

[0119] 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-pyridylmethyl, 2-quinolinylmethyl, 2-pyridylmethyl N-oxonium, 9-anthraylmethyl, 9-fluorenylmethyl, xanthyl, ferroceneylmethyl, diphenylmethyl, bis(4-methoxyphenyl)methyl, 5-dibenzosuccinyl, triphenylmethyl, diphenyl-4-pyridylmethyl, phenyl, 2,4-dinitrophenyl, tert-butyl, 1-adamantyl, methoxymethyl (MOM), isobutoxymethyl, benzyloxymethyl, 2-tetrahydropyranyl, benzylthiomethyl, phenylthiomethyl, thiazolidinyl, acetaminomethyl, trimethylacetamidemethyl, benzamidemethyl. Allyloxycarbonylaminomethyl, phenylacetamide methyl, phthalimide methyl, acetylmethyl, carboxymethyl, cyanomethyl, (2-nitro-1-phenyl)ethyl, 2-(2,4-dinitrophenyl)ethyl, 2-cyanoethyl, 2-(trimethylsilyl)ethyl, 2,2-bis(ethoxy)ethyl, (1-m-nitrophenyl-2-benzoyl)ethyl, 2-benzenesulfonylethyl, 2-(4-methylbenzenesulfonyl)-2-methylpropyl-2-yl, acetyl, benzoyl, trifluoroacetyl, N-[[(p-biphenyl)isopropoxy]carbonyl]-N-methyl]-γ-aminothiobutyrate, 2,2,2-trichloroethoxycarbonyl, tert-butoxycarbonyl, benzyloxycarbonyl, p-methoxybenzyloxycarbonyl, N-ethyl, N-methoxymethyl, sulfonate, thionyl carbonate, 3-nitro-2-pyridine thiosulfate, oxythiophene.

[0120] The compounds of the present invention

[0121] Liposome-based mediators are considered attractive carriers for therapeutics and require continued development efforts. While liposome-based mediators incorporating certain lipid components have shown promising results in terms of encapsulation, stability, and site localization, there remains a significant need to improve liposome-based delivery systems. For example, significant drawbacks of liposome delivery systems involve the construction of liposomes with sufficient cell culture or in vivo stability to achieve the desired target cells and / or intracellular compartments, and the ability of such liposome delivery systems to efficiently release their encapsulated substances into such target cells.

[0122] Specifically, there remains a need for improved lipid compounds that exhibit enhanced pharmacokinetic properties and can deliver macromolecules (such as nucleic acids) to a variety of cell types and tissues with increased efficiency. Importantly, there remains a particular need for novel lipid compounds characterized by reduced toxicity and the ability to efficiently deliver encapsulated nucleic acids and polynucleotides to target cells, tissues, and organs.

[0123] This article describes a novel class of cationic lipid compounds for improved in vivo delivery of therapeutic agents such as nucleic acids. Specifically, the cationic lipids described herein can optionally be used in conjunction with other lipids to formulate lipid-based nanoparticles (e.g., liposomes) for encapsulating therapeutic agents, such as nucleic acids (e.g., DNA, siRNA, mRNA, microRNA) for therapeutic purposes.

[0124] In the embodiments, the compounds of the present invention as described herein may provide one or more desired properties or characteristics. That is, in some embodiments, the compounds of the present invention described herein may be characterized as having one or more properties that provide advantages over other similarly classified lipids. For example, the compounds disclosed herein may allow control and customization of the properties of liposome compositions (e.g., lipid nanoparticles) in which they are components. Specifically, the compounds disclosed herein may be characterized by enhanced transfection efficiency and their ability to induce specific biological outcomes. Such outcomes may include, for example, enhanced cellular uptake, endosome / lysosome disruption, and / or promotion of the release of intracellular encapsulated materials (e.g., polynucleotides). In addition, the compounds disclosed herein have favorable pharmacokinetic properties, biodistribution, and efficiency (e.g., due to the different dissociation rates of the polymer groups used).

[0125] This application demonstrates that the cationic lipids of the present invention can not only be synthesized from readily available starting materials, but also possess unexpectedly high encapsulation efficiency.

[0126] Furthermore, the cationic lipids of the present invention have cleavable groups, such as ester groups and disulfides. These cleavable groups (e.g., esters and disulfides) are considered to improve biodegradability and thus contribute to their advantageous toxicity characteristics.

[0127] The compounds of the present invention

[0128] This document provides compounds that are cationic lipids. For example, the cationic lipids of the present invention comprise compounds having a structure according to formula (I):

[0129]

[0130] Where L1 is a bond, (C1-C6)alkyl or (C2-C6)alkenyl;

[0131] Where X is O or S;

[0132] Where R 1 R 2 R 3 R 4 and R 5 Each is independently selected from H, OH, optionally substituted (C1-C6)alkyl, optionally substituted (C2-C6)alkenyl, optionally substituted (C2-C6)ynyl, optionally substituted (C1-C6)alkoxy and -OC(O)R';

[0133] Where R 1 R 2 R 3 R 4 or R 5 At least one of them is -OC(O)R';

[0134] Where R' is

[0135]

[0136] Where R 6 for

[0137]

[0138] Where m and p are each independently 0, 1, 2, 3, 4 or 5;

[0139] Where R 7 Selected from H, optionally substituted (C1-C6)alkyl, optionally substituted (C2-C6)alkenyl, optionally substituted (C2-C6)ynyl, optionally substituted (C1-C6)acyl, -(CH2). k R A Or -(CH2) k CH(OR 11 )R A ;

[0140] Where R 8 Selected from H, optionally substituted (C1-C6)alkyl, optionally substituted (C2-C6)alkenyl, optionally substituted (C2-C6)ynyl, optionally substituted (C1-C6)acyl, -(CH2). n R B Or -(CH2) n CH(OR 12 )R B ;

[0141] Where R 9Selected from H, optionally substituted (C1-C6)alkyl, optionally substituted (C2-C6)alkenyl, optionally substituted (C2-C6)ynyl, optionally substituted (C1-C6)acyl, -(CH2). q R C Or -(CH2) q CH(OR 13 )R C ;

[0142] Where R 10 Selected from H, optionally substituted (C1-C6)alkyl, optionally substituted (C2-C6)alkenyl, optionally substituted (C2-C6)ynyl, optionally substituted (C1-C6)acyl, -(CH2). r R D Or -(CH2) r CH(OR 14 )R D ;

[0143] Where k, n, q and r are each 1, 2, 3, 4 or 5 independently;

[0144] Or one of them (i)R 7 and R 8 Or (ii)R 9 and R 10 Together they form optionally substituted 5- or 6-membered heterocyclic alkyl or heteroaryl groups, wherein the heterocyclic alkyl or heteroaryl group comprises 1 to 3 heteroatoms selected from N, O and S;

[0145] Where R 11 R 12 R 13 and R 14 Each is independently selected from H, methyl, ethyl, or propyl;

[0146] Where R A R B R C and R D Each is independently selected from the arbitrarily substituted (C6-C) 20 )alkyl, optionally substituted (C6-C 20 alkenyl, optionally substituted (C6-C) 20 ) ynyl group, optionally substituted (C6-C 20 Acyl, optionally substituted -OC(O)alkyl, optionally substituted -OC(O)alkenyl, optionally substituted (C1-C6)monoalkylamino, optionally substituted (C1-C6)dialkylamino, optionally substituted (C1-C6)alkoxy, -OH, -NH2;

[0147] Where R 7 R 8 R9 R 10 At least one of them includes R A R B R C or R D Part, wherein the R A R B R C or R D Independently selected from optional substitutions (C6-C) 20 )alkyl, optionally substituted (C6-C 20 alkenyl, optionally substituted (C6-C) 20 ) ynyl group, optionally substituted (C6-C 20 Acyl group, optionally substituted -OC(O)(C6-C 20 )alkyl or optionally substituted -OC(O)(C6-C 20 alkenyl;

[0148] Or its pharmaceutically acceptable salt.

[0149] In the embodiments, any alkyl, alkenyl, alkynyl, acyl, alkoxy, monoalkylamino, dialkylamino, heterocyclic alkyl, or heteroaryl group is optionally substituted by one or more substituents selected from the group consisting of: (C1-C6)alkyl, (C2-C6)alkenyl, (C2-C6)alkynyl, (C1-C6)acyl, (C1-C6)alkoxy, halogen, -COR, -CO2H, -CO2R, -CN, -OH, -OR, -OCOR, -OCO2R, -NH2, -NHR, -N(R)2, -SR, or -SO2R, or the two twin hydrogens on the carbon atom are substituted by the group =NH, wherein each instance of R is independently a C1-C10 aliphatic alkyl group.

[0150] In this embodiment, L1 is independently the key.

[0151] In the examples, L1 is a (C1-C6) alkyl group.

[0152] In the embodiments, L1 is (C2-C6) alkenyl.

[0153] In the examples, L1 is C2 alkenyl.

[0154] In this embodiment, RA and RB are the same. In this embodiment, RC and RD are the same. In this embodiment, RA and RB are the same, and RC and RD are the same.

[0155] In this embodiment, RA and RB are different. In this embodiment, RC and RD are different. In this embodiment, RA and RB are different, and RC and RD are different.

[0156] In the embodiments, RA, RB, RC, and RD are the same.

[0157] In the embodiments, RA, RB, RC, and RD are different.

[0158] In the embodiments, RA, RB, RC or RD are each independently selected from optionally substituted (C6-C20)alkyl, optionally substituted (C6-C20)alkenyl, optionally substituted (C6-C20)ynyl, optionally substituted (C6-C20)acyl, optionally substituted -OC(O)(C6-C20)alkyl or optionally substituted -OC(O)(C6-C20)alkenyl.

[0159] In the embodiments, RA, RB, RC, or RD are the same and selected from optionally substituted (C6-C20)alkyl, optionally substituted (C6-C20)alkenyl, optionally substituted (C6-C20)ynyl, optionally substituted (C6-C20)acyl, optionally substituted -OC(O)(C6-C20)alkyl, or optionally substituted -OC(O)(C6-C20)alkenyl.

[0160] In the embodiments, RA and RB are each independently selected from optionally substituted (C6-C20) alkyl, optionally substituted (C6-C20) alkenyl, or optionally substituted (C6-C20) ynyl.

[0161] In the examples, RA and RB are the same and are selected from optionally substituted (C6-C20) alkyl, optionally substituted (C6-C20) alkenyl, and optionally substituted (C6-C20) ynyl.

[0162] In the examples, RA and RB are each independently an optionally substituted (C6-C20) alkyl group.

[0163] In the examples, RA and RB are identical and are optionally substituted (C6-C20) alkyl groups.

[0164] In the embodiments, RA and RB are each independently an optionally substituted (C6-C20) alkenyl group.

[0165] In the examples, RA and RB are identical and are optionally substituted (C6-C20) alkenyl groups.

[0166] In the embodiments, RA and RB are each independently an optionally substituted (C6-C20) ynyl group.

[0167] In the embodiments, RA and RB are identical and are optionally substituted (C6-C20) ynyl groups.

[0168] In the embodiments, RA and RB are each independently an optionally substituted (C6-C20) acyl group.

[0169] In the embodiments, RA and RB are identical and are optionally substituted (C6-C20) acyl groups.

[0170] In the embodiments, RA and RB are each independently an optionally substituted -OC(O)(C6-C20) alkyl group.

[0171] In the examples, RA and RB are identical and are optionally substituted -OC(O)(C6-C20) alkyl groups.

[0172] In the embodiments, RA and RB are each independently an optionally substituted -OC(O)(C6-C20) alkenyl group.

[0173] In the embodiments, RA and RB are identical and are optionally substituted -OC(O)(C6-C20) alkenyl groups.

[0174] In the embodiment, R7 = -(CH2)kCH(OR11)RA, R8 is -(CH2)nCH(OR12)RB, and RA and RB are each independently selected from:

[0175]

[0176] In the embodiment, R7 = -(CH2)kCH(OR11)RA, R8 is -(CH2)nCH(OR12)RB, and RA and RB are the same and selected from:

[0177]

[0178]

[0179] In the embodiment, R7 = -(CH2)kCH(OR11)RA, R8 is -(CH2)nCH(OR12)RB, and both RA and RB are C8H17.

[0180] In the embodiment, R7 = -(CH2)kCH(OR11)RA, R8 is -(CH2)nCH(OR12)RB, and both RA and RB are C10H21.

[0181] In the embodiment, R7 = -(CH2)kCH(OR11)RA, R8 is -(CH2)nCH(OR12)RB, and both RA and RB are C12H25.

[0182] In the embodiment, R7 = -(CH2)kCH(OR11)RA, R8 is -(CH2)nCH(OR12)RB, and both RA and RB are...

[0183] In this embodiment, X is 0.

[0184] In this embodiment, X is S.

[0185] In the embodiment, only one of R1, R2, R3, R4, and R5 is -OC(O)R'. In the embodiment, only one of R1, R2, R3, R4, and R5 is -OC(O)R', and none of R1, R2, R3, R4, or R5 is OH.

[0186] In the embodiment, two of R1, R2, R3, R4, and R5 are -OC(O)R'. In the embodiment, two of R1, R2, R3, R4, and R5 are -OC(O)R', and none of R1, R2, R3, R4, or R5 are OH.

[0187] In the embodiment, three of R1, R2, R3, R4 and R5 are -OC(O)R'.

[0188] In this embodiment, R1 is -OC(O)R'. In this embodiment, R5 is -OC(O)R'. In this embodiment, both R1 and R5 are -OC(O)R'.

[0189] In this embodiment, R2 is -OC(O)R'. In this embodiment, R4 is -OC(O)R'. In this embodiment, both R2 and R4 are -OC(O)R'.

[0190] In the embodiment, R3 is -OC(O)R'.

[0191] In the embodiment, R3 is -OC(O)R' and R2 is OMe.

[0192] In the embodiment, L1 is a bond, R3 is -OC(O)R', and R2 is OMe.

[0193] In the embodiment, R3 is -OC(O)R', and R2 and R4 are OMe.

[0194] In the embodiment, L1 is a bond, R3 is -OC(O)R', and R2 and R4 are OMe.

[0195] In the embodiments, L1 is (C2-C6) alkenyl, R3 is -OC(O)R', and R2 and R4 are OMe.

[0196] In the examples, L1 is C2-alkenyl, R3 is -OC(O)R', and R2 and R4 are OMe.

[0197] In the embodiment, R7 is -(CH2)kCH(OR11)RA.

[0198] In the embodiment, R7 is -(CH2)1CH(OR11)RA.

[0199] In the embodiment, R7 is -(CH2)1CH(OH)RA.

[0200] In the embodiment, R8 is -(CH2)nCH(OR12)RB.

[0201] In the embodiment, R8 is -(CH2)1CH(OR12)RB.

[0202] In the embodiment, R8 is -(CH2)1CH(OH)RB.

[0203] In the embodiment, R7 is -(CH2)kCH(OR11)RA and R8 is -(CH2)nCH(OR12)RB.

[0204] In the embodiment, R7 is -(CH2)1CH(OR11)RA and R8 is -(CH2)1CH(OR12)RB.

[0205] In the embodiments, R7 is -(CH2)1CH(OH)RA and R8 is -(CH2)1CH(OH)RB.

[0206] In the embodiments, R7 and R8 are each optionally substituted (C1-C6) alkyl groups, for example, -CO2R aa Substituted (C1-C6) alkyl groups, wherein R aa For C1-C 50 Alkyl group. In the examples, R 7 and R 8 Each for itself - CO2R aa Substituted (C1-C6) alkyl groups, wherein R aa C1-C 40 Alkyl group. In the examples, R 7 and R 8 Each for itself - CO2R aa Substituted (C1-C6) alkyl groups, wherein R aa C1-C 30 Alkyl group. In the examples, R 7 and R 8 Each for itself - CO2R aa Substituted (C1-C6) alkyl groups, wherein R aa C1-C 20 alkyl.

[0207] In the embodiments, R7 and R8 are identical and each is an optionally substituted (C1-C6) alkyl group, for example, a (C1-C6) alkyl group substituted with -CO2Raa, wherein Raa is a C1-C50 alkyl group. In the embodiments, R7 and R8 are identical and each is a (C1-C6) alkyl group substituted with -CO2Raa, wherein Raa is a C1-C40 alkyl group. In the embodiments, R7 and R8 are identical and each is a (C1-C6) alkyl group substituted with -CO2Raa, wherein Raa is a C1-C30 alkyl group. In the embodiments, R7 and R8 are each a (C1-C6) alkyl group substituted with -CO2Raa, wherein Raa is a C1-C20 alkyl group.

[0208] In the embodiment, R7 and R8 are each

[0209] In the embodiment, R7 and R8 are each

[0210] In the embodiments, R9 and R10 are each independently selected from H, optionally substituted (C1-C6) alkyl, optionally substituted (C2-C6) alkenyl, and optionally substituted (C2-C6) ynyl.

[0211] In the embodiments, R9 and R10 are each independently an optionally substituted (C1-C6) alkyl or an optionally substituted (C2-C6) alkenyl.

[0212] In the examples, R9 and R10 are both optionally substituted (C1-C6) alkyl or optionally substituted (C2-C6) alkenyl groups.

[0213] In the examples, R9 and R10 are both optionally substituted (C1-C6) alkyl groups.

[0214] In the embodiment, both R9 and R10 are -CH3.

[0215] In the embodiment, R7 is -(CH2)kCH(OR11)RA, R8 is -(CH2)nCH(OR12)RB, and R9 and R10 are both -CH3.

[0216] In the embodiment, R7 is -(CH2)1CH(OR11)RA, R8 is -(CH2)1CH(OR12)RB, and R9 and R10 are both -CH3.

[0217] In the embodiments, R7 is -(CH2)1CH(OH)RA, R8 is -(CH2)1CH(OH)RB, and R9 and R10 are both -CH3.

[0218] In the embodiment, R7 is -(CH2)kCH(OR11)RA, R8 is -(CH2)nCH(OR12)RB, RA and RB are both C8H17, and R9 and R10 are both -CH3.

[0219] In the embodiments, R7 is -(CH2)1CH(OH)RA, R8 is -(CH2)1CH(OH)RB, RA and RB are both C8H17, and R9 and R10 are both -CH3.

[0220] In the embodiment, R7 is -(CH2)kCH(OR11)RA, R8 is -(CH2)nCH(OR12)RB, RA and RB are both C10H21, and R9 and R10 are both -CH3.

[0221] In the embodiments, R7 is -(CH2)1CH(OH)RA, R8 is -(CH2)1CH(OH)RB, RA and RB are both C10H21, and R9 and R10 are both -CH3.

[0222] In the embodiment, R7 is -(CH2)kCH(OR11)RA, R8 is -(CH2)nCH(OR12)RB, RA and RB are both C12H25, and R9 and R10 are both -CH3.

[0223] In the embodiments, R7 is -(CH2)1CH(OH)RA, R8 is -(CH2)1CH(OH)RB, RA and RB are both C12H25, and R9 and R10 are both -CH3.

[0224] In the embodiment, R7 is -(CH2)kCH(OR11)RA, R8 is -(CH2)nCH(OR12)RB, RA and RB are both C16H29, and R9 and R10 are both -CH3.

[0225] In the embodiments, R7 is -(CH2)1CH(OH)RA, R8 is -(CH2)1CH(OH)RB, RA and RB are both C16H29, and R9 and R10 are both -CH3.

[0226] In one embodiment, p, g, and r are the same. In another embodiment, one or more of p, q, and r are different. In another embodiment, q and r are the same, and p is different. In another embodiment, p and q are the same, and r is different. In another embodiment, p and r are the same, and q is different. In yet another embodiment, p, q, and r are different.

[0227] In one embodiment, k, m, and n are the same. In another embodiment, one or more of k, m, and n are different. In another embodiment, k and m are the same, and n is different. In another embodiment, m and n are the same, and k is different. In another embodiment, k and n are the same, and m is different. In yet another embodiment, k, m, and n are different.

[0228] In this embodiment, m is 1, 2, 3, 4, or 5. In this embodiment, m is 0. In this embodiment, m is 1. In this embodiment, m is 2. In this embodiment, m is 3. In this embodiment, m is 4. In this embodiment, m is 5. In this embodiment, m is 0, 1, 2, 3, or 4.

[0229] In the embodiments, p is 1, 2, 3, 4, or 5. In the embodiments, p is 0. In the embodiments, p is 1. In the embodiments, p is 2. In the embodiments, p is 3. In the embodiments, p is 4. In the embodiments, p is 5. In the embodiments, p is 0, 1, 2, 3, or 4.

[0230] In the embodiment, m is 2 and p is 2.

[0231] In the embodiment, m is 3 and p is 2.

[0232] In the embodiment, k and n = 1, and m = 2.

[0233] In the embodiment, k and n = 1, and m = 3.

[0234] In the embodiment, q and r = 1, and p = 2.

[0235] In the embodiment, k, n, q and r are each 1, m is 2 or 3, and p is 2.

[0236] In the embodiment, R' is:

[0237]

[0238] In the embodiment, R' is And k and n = 1, and m = 2 or 3.

[0239] In the embodiment, R' is And k and n = 1, and m = 2.

[0240] In the embodiment, R' is And k and n = 1, and m = 3.

[0241] In the embodiment, R' is And R 11 and R 12 For H.

[0242] In the embodiment, R' is And k and n = 1, m = 2 or 3, and R 11 and R 12 For H.

[0243] In the embodiment, R' is And k and n = 1, m = 2, and R 11 and R 12 For H.

[0244] In the embodiment, R' is And k and n = 1, m = 3, and R 11 and R 12 For H.

[0245] In the embodiment, R' is:

[0246]

[0247] R' has the following structure In any of the above embodiments, R A and R B It can be defined as in any of paragraphs

[0104] to

[0129] .

[0248] In the embodiment, R 6 for:

[0249]

[0250] In the embodiment, R 6 for q and r = 1, and p = 2.

[0251] In the embodiment, R 6 for And R 13 and R 14 For H.

[0252] In the embodiment, R 6 for q and r = 1, p = 2, and R 13 and R 14 For H.

[0253] In the embodiment, R 6 Choose from the following groups:

[0254]

[0255] In the embodiment, R 6 Choose from the following groups:

[0256] In the embodiment, R 6 Choose from the following groups:

[0257] In the embodiment, R 6 for:

[0258]

[0259] In the embodiment, R 6 for:

[0260]

[0261] In the embodiment, R 6 for:

[0262]

[0263] In the embodiment, R 6 Choose from the following groups:

[0264] In the embodiment, R 6 for:

[0265]

[0266] In the embodiment, R 6 for:

[0267]

[0268] In the embodiment, R 6 for:

[0269]

[0270] In the embodiment, R 6 for:

[0271]

[0272] In the embodiment, R 6 for:

[0273]

[0274] In the embodiment, R 6 Same as R'.

[0275] In the embodiment, R 6 for R' is m is 2, and p is 2.

[0276] In the embodiment, R 6 for R' is m is 3 and p is 2.

[0277] In the embodiment, L1 key, R 3 For -OC(O)R', R 2 and R 4 For OMe, R 6 for And R' is

[0278] In the embodiment, L1 is the key, R 3 For -OC(O)R', R 2 and R 4 For OMe, R 6 for And R' is

[0279] In this embodiment, X = 0, L1 is the bond, and R 3 For -OC(O)R', R 2 and R 4 For OMe, R 6 for And R' is

[0280] In this embodiment, X = 0, L1 is the bond, and R 3 For -OC(O)R', R 2 and R 4 For OMe, R 1 and R 5 For H, R 6 for And R' is

[0281] In this embodiment, X = 0, L1 is the bond, and R 3 For -OC(O)R', R 2 and R 4 For OMe, R 6 for And R' is

[0282] In this embodiment, X = 0, L1 is the bond, and R 3 For -OC(O)R', R 2 and R 4 For OMe, R 1 and R 5 For H, R 6 for And R' is

[0283] In the embodiment, L1 is the key, R 3 For -OC(O)R', R 2 For OMe, R 6 for And R' is

[0284] In this embodiment, X = 0, L1 is the bond, and R 3 For -OC(O)R', R 2 For OMe, R 6 for And R' is

[0285] In this embodiment, X = 0, L1 is the bond, and R 3 For -OC(O)R', R 2 For OMe, R 1 R 4 and R 5 For H, R 6 for And R' is

[0286] In the embodiment, L1 is the key, R 3 For -OC(O)R', R 2 For OMe, R 6 for And R' is

[0287] In this embodiment, X = 0, L1 is the bond, and R 3 For -OC(O)R', R 2 For OMe, R 6 for And R' is

[0288] In this embodiment, X = 0, L1 is the bond, and R 3 For -OC(O)R', R 2 For OMe, R 1 R 4 and R 5 For H, R 6 for And R' is

[0289] In the examples, L1 is C2 alkenyl, R 3 For -OC(O)R', R 2 and R 4 For OMe, R 6 for And R' is

[0290] In the examples, L1 is C2 alkenyl, R 3 For -OC(O)R', R 2 and R 4 For OMe, R 6 for And R' is

[0291] In the embodiments, X = O, L1 is C2 alkenyl, and R 3 For -OC(O)R', R 2 and R 4 For OMe, R 6 for And R ' for

[0292] In the embodiments, X = O, L1 is C2 alkenyl, and R 3 For -OC(O)R', R 2 and R 4 For OMe, R 1 and R 5 For H, R 6 for And R' is

[0293] In the embodiments, X = O, L1 is C2 alkenyl, and R 3 For -OC(O)R', R 2 and R 4 For OMe, R 6 for And R' is

[0294] In the embodiments, X = O, L1 is C2 alkenyl, and R 3 For -OC(O)R', R 2 and R 4 For OMe, R 1 and R 5 For H, R 6 for And R' is

[0295] In the embodiment, R 6 for R' is R 7 and R 8 Each for itself - CO2R aa Substituted (C1-C6) alkyl groups, wherein R aa For C1-C 50 alkyl.

[0296] In the embodiment, R 6 for R' is R 7 and R 8 Each for itself - CO2R aa Substituted (C1-C6) alkyl groups, wherein R aa For C1-C 50 Alkyl group, m is 2, and p is 2. In some embodiments, R 7 and R 8 Same. In some embodiments, L1 is the key, R 3 For -OC(O)R', R 2 and R 4 For OMe.

[0297] In the embodiment, L1 is the key, R 3 For -OC(O)R', R 2 and R 4 For OMe, R 6 for R' is And R 7 and R 8 Each for itself - CO2R aa Substituted (C1-C6) alkyl groups, wherein R aa For C1-C 50 Alkyl group. In some embodiments, R 7 and R 8 same.

[0298] In the embodiment, L1 is the key, R 3 For -OC(O)R', R 2 and R 4 For OMe, R 6 for R' is And R 7 and R 8 Each for itself - CO2R aa Substituted (C1-C6) alkyl groups, wherein R aa For C1-C 50 Alkyl group, and m is 2. In some embodiments, R 7 and R 8 same.

[0299] In this embodiment, X = 0, L1 is the bond, and R 3 For -OC(O)R', R 2 and R 4 For OMe, R 6 for R' for And R 7 and R 8 Each for itself - CO2R aa Substituted (C1-C6) alkyl groups, wherein R aa For C1-C 50 Alkyl group. In some embodiments, R 7 and R 8 Same. In some embodiments, m is 2.

[0300] In this embodiment, X = 0, L1 is the bond, and R 3 For -OC(O)R', R 2 and R 4 For OMe, R 1 and R 5 For H, R 6 for R' is And R 7 and R 8 Each for itself - CO2R aa Substituted (C1-C6) alkyl groups, wherein R aa For C1-C 50 Alkyl group. In some embodiments, R 7 and R 8 Same. In some embodiments, m is 2.

[0301] In this embodiment, X = 0, L1 is the bond, and R 3 For -OC(O)R', R 2 and R 4 For OMe, R 6 for R' is And R 7 and R 8 Each for itself - CO2R aa Substituted (C1-C6) alkyl groups, wherein R aa For C1-C 50 Alkyl group, and m is 2. In some embodiments, R 7 and R 8 same.

[0302] In this embodiment, X = 0, L1 is the bond, and R 3 For -OC(O)R', R 2 and R 4 For OMe, R 1 and R 5 For H, R 6 for R' is R 7 and R8 Each for itself - CO2R aa Substituted (C1-C6) alkyl groups, wherein R aa For C1-C 50 Alkyl group, and m is 2. In some embodiments, R 7 and R 8 same.

[0303] In the embodiment, L1 is the key, R 3 For -OC(O)R', R 2 For OMe, R 6 for R' is And R 7 and R 8 Each for itself - CO2R aa Substituted (C1-C6) alkyl groups, wherein R aa For C1-C 50 Alkyl group. In some embodiments, R 7 and R 8 Same. In some embodiments, m is 2.

[0304] In this embodiment, X = 0, L1 is the bond, and R 3 For -OC(O)R', R 2 For OMe, R 6 for R' is And R 7 and R 8 Each for itself - CO2R aa Substituted (C1-C6) alkyl groups, wherein R aa For C1-C 50 Alkyl group. In some embodiments, R 7 and R 8 Same. In some embodiments, m is 2.

[0305] In this embodiment, X = 0, L1 is the bond, and R 3 For -OC(O)R', R 2 For OMe, R 1 R 4 and R 5 For H, R 6 for R' is And R 7 and R 8 Each for itself - CO2R aa Substituted (C1-C6) alkyl groups, wherein R aa For C1-C 50 Alkyl group. In some embodiments, R 7 and R8 Same. In some embodiments, m is 2.

[0306] In the embodiment, L1 is the key, R 3 For -OC(O)R', R 2 For OMe, R 6 for R' is And R 7 and R 8 Each for itself - CO2R aa Substituted (C1-C6) alkyl groups, wherein R aa For C1-C 50 Alkyl group, and m is 2. In some embodiments, R 7 and R 8 same.

[0307] In this embodiment, X = 0, L1 is the bond, and R 3 For -OC(O)R', R 2 For OMe, R 1 R 4 and R 5 For H, R 6 for R' is R 7 and R 8 Each for itself - CO2R aa Substituted (C1-C6) alkyl groups, wherein R aa For C1-C 50 Alkyl group, and m is 2. In some embodiments, R 7 and R 8 same.

[0308] In the examples, L1 is C2 alkenyl, R 3 For -OC(O)R', R 2 and R 4 For OMe, R 6 for R' is And R 7 and R 8 Each for itself - CO2R aa Substituted (C1-C6) alkyl groups, wherein R aa For C1-C 50 Alkyl group. In some embodiments, R 7 and R 8 same.

[0309] In the examples, L1 is C2 alkenyl, R 3 For -OC(O)R', R 2 and R 4For OMe, R 6 for R' is And R 7 and R 8 Each for itself - CO2R aa Substituted (C1-C6) alkyl groups, wherein R aa For C1-C 50 Alkyl group, and m is 2. In some embodiments, R 7 and R 8 same.

[0310] In the embodiments, X = O, L1 is C2 alkenyl, and R 3 For -OC(O)R', R 2 and R 4 For OMe, R 6 for R' is And R 7 and R 8 Each for itself - CO2R aa Substituted (C1-C6) alkyl groups, wherein R aa For C1-C 50 Alkyl group. In some embodiments, R 7 and R 8 same.

[0311] In the embodiments, X = O, L1 is C2 alkenyl, and R 3 For -OC(O)R',R 2 and R 4 For OMe, R 1 and R 5 For H, R 6 for R ' for And R 7 and R 8 Each for itself - CO2R aa Substituted (C1-C6) alkyl groups, wherein R aa For C1-C 50 Alkyl group. In some embodiments, R 7 and R 8 same.

[0312] In the embodiments, X = O, L1 is C2 alkenyl, and R 3 For -OC(O)R', R 2 and R 4 For OMe, R 6 for R ' for R 7 and R8 Each for itself - CO2R aa Substituted (C1-C6) alkyl groups, wherein R aa For C1-C 50 Alkyl group, and m is 2. In some embodiments, R 7 and R 8 same.

[0313] In the embodiments, X = O, L1 is C2 alkenyl, and R 3 For -OC(O)R',R 2 and R 4 For OMe, R 1 and R 5 For H, R 6 for R ' for R 7 and R 8 Each for itself - CO2R aa Substituted (C1-C6) alkyl groups, wherein R aa For C1-C 50 Alkyl group, and m is 2. In some embodiments, R 7 and R 8 same.

[0314] In which R 7 and R 8 For -CO2R aa Substituted (C1-C6) alkyl groups, wherein R aa For C1-C 50 In any of the above embodiments of alkyl groups, R aa C1-C can be used as an alternative. 40 alkyl.

[0315] In which R 7 and R 8 For -CO2R aa Substituted (C1-C6) alkyl groups, wherein R aa For C1-C 50 In any of the above embodiments of alkyl groups, R aa C1-C can be used as an alternative. 30 alkyl.

[0316] In which R 7 and R 8 For -CO2R aa Substituted (C1-C6) alkyl groups, wherein R aa For C1-C 50 In any of the above embodiments of alkyl groups, R aa C1-C can be used as an alternative.20 alkyl.

[0317] In which R 7 and R 8 For -CO2R aa Substituted (C1-C6) alkyl groups, wherein R aa For C1-C 50 In any of the embodiments of alkyl groups, R 7 and R 8 Each can do it themselves

[0318] In which R 7 and R 8 Each for itself - CO2R aa Substituted (C1-C6) alkyl groups, wherein R aa For C1-C 50 In any of the embodiments of alkyl groups, R 7 and R 8 Each can do it themselves

[0319] In embodiments, the cationic lipids of the present invention comprise compounds having the structure according to formula (II):

[0320]

[0321] Or a pharmaceutically acceptable salt thereof, wherein R 1 -R 6 And X as defined in this article.

[0322] In embodiments, the cationic lipids of the present invention comprise compounds having a structure according to formula (IIA):

[0323]

[0324] Or a pharmaceutically acceptable salt thereof, wherein R', R 6 And X as defined in this article.

[0325] In embodiments, the cationic lipids of the present invention comprise compounds having a structure according to formula (IIB), (IIC), (IID), (IIE), (IIJ), or (IIK):

[0326]

[0327]

[0328]

[0329] Or its pharmaceutically acceptable salt.

[0330] In the embodiments, the cationic lipid of the present invention has the following structure:

[0331] Or its pharmaceutically acceptable salt.

[0332] In the embodiments, the cationic lipid of the present invention has the following structure:

[0333] Or its pharmaceutically acceptable salt.

[0334] In the embodiments, the cationic lipid of the present invention has the following structure:

[0335]

[0336] Or its pharmaceutically acceptable salt.

[0337] In the embodiments, the cationic lipid of the present invention has the following structure:

[0338]

[0339] Or its pharmaceutically acceptable salt.

[0340] In embodiments, the cationic lipids of the present invention comprise compounds having a structure according to formula (IIF):

[0341]

[0342] Or a pharmaceutically acceptable salt thereof, wherein R', R 6 And X as defined in this article.

[0343] In embodiments, the cationic lipids of the present invention comprise compounds having a structure according to formula (IIG):

[0344]

[0345] Or a pharmaceutically acceptable salt thereof, wherein R', R 6 And X as defined in this article.

[0346] In embodiments, the cationic lipids of the present invention comprise compounds having a structure according to formula (IIH):

[0347]

[0348] Where one of Y and Z is OH and the other is -OC(O)R', or where Y and Z are each independently -OC(O)R', and where R' and R 6 And X as defined in this article.

[0349] In embodiments, the cationic lipids of the present invention comprise compounds having a structure according to formula (III):

[0350]

[0351] Or a pharmaceutically acceptable salt thereof, wherein R 1 -R 6 And X as defined in this article.

[0352] In embodiments, the cationic lipids of the present invention comprise compounds having a structure according to formula (IIIA):

[0353]

[0354] Or a pharmaceutically acceptable salt thereof, wherein R', R 6 And X as defined in this article.

[0355] In embodiments, the cationic lipids of the present invention comprise compounds having a structure according to formula (IIIB):

[0356]

[0357] Or a pharmaceutically acceptable salt thereof, wherein R A R B p is as defined in this paper.

[0358] In the embodiments, the cationic lipid of the present invention has the following structure:

[0359]

[0360] Or a pharmaceutically acceptable salt thereof, wherein R A and R B As defined in this article.

[0361] In the embodiments, the cationic lipid of the present invention has the following structure:

[0362]

[0363] Or a pharmaceutically acceptable salt thereof, wherein R A and R B As defined in this article.

[0364] In the embodiments, the cationic lipid of the present invention has the following structure:

[0365]

[0366] Or its pharmaceutically acceptable salt.

[0367] In the embodiments, the cationic lipid of the present invention has the following structure:

[0368]

[0369] Or its pharmaceutically acceptable salt.

[0370] In embodiments, the cationic lipids of the present invention comprise compounds having a structure according to formula (IIID):

[0371]

[0372] Or a pharmaceutically acceptable salt thereof, wherein R', R 6 And X as defined in this article.

[0373] In embodiments, the cationic lipids of the present invention comprise compounds having a structure according to formula (IIIE), (IIIF), (IIIG), (IIIH), (IIII), (IIIJ), or (IIIK):

[0374]

[0375]

[0376]

[0377] Or its pharmaceutically acceptable salt.

[0378] In the embodiments, the cationic lipid of the present invention has the following structure:

[0379]

[0380] In the embodiments, the cationic lipid of the present invention has the following structure:

[0381]

[0382] In the embodiments, the cationic lipid of the present invention has the following structure:

[0383]

[0384] In the embodiments, the cationic lipid of the present invention has the following structure:

[0385]

[0386] In the embodiments, the cationic lipid of the present invention has the following structure:

[0387]

[0388] In the embodiments, the cationic lipid of the present invention has the following structure:

[0389]

[0390] In the embodiments, the cationic lipid of the present invention has the following structure:

[0391]

[0392] In embodiments, the cationic lipids of the present invention comprise compounds having a structure according to formula (IIIL):

[0393]

[0394] Or a pharmaceutically acceptable salt thereof, wherein R', R 6 And X as defined in this article.

[0395] In embodiments, the cationic lipids of the present invention comprise compounds having the structure according to formula (IV):

[0396]

[0397] Where M is selected from H, OH, OMe, or Me.

[0398] Or a pharmaceutically acceptable salt thereof, wherein R A R B m and p are as defined in this paper.

[0399] In the embodiments, the cationic lipids of the present invention comprise compounds having a structure according to formula (VI), (VII), (VIII), (IX) or (X):

[0400]

[0401]

[0402] Or its pharmaceutically acceptable salt.

[0403] Where one of Y and Z is OH and the other is -OC(O)R', or where Y and Z are each independently -OC(O)R', and where R ' R 6 And X as defined herein, or its pharmaceutically acceptable salt.

[0404] In the embodiment, one of Y and Z is OH and the other is -OC(O)R'.

[0405] In the embodiments, Y is OH and Z is -OC(O)R'.

[0406] In the embodiments, Y is -OC(O)R' and Z is OH.

[0407] In the embodiment, both Y and Z are -OC(O)R'.

[0408] In an embodiment, a composition is provided comprising any one of the cationic lipids described in the foregoing embodiments, one or more non-cationic lipids, one or more cholesterol-based lipids, and one or more PEG-modified lipids. In an embodiment, this composition is lipid nanoparticles. In an embodiment, one or more cationic lipids constitute about 30 mol%-60 mol% of the lipid nanoparticles. In an embodiment, one or more non-cationic lipids constitute 10 mol%-50 mol% of the lipid nanoparticles. In an embodiment, one or more PEG-modified lipids constitute 1 mol%-10 mol% of the lipid nanoparticles. In an embodiment, cholesterol-based lipids constitute 10 mol%-50 mol% of the lipid nanoparticles. In an embodiment, the lipid nanoparticles encapsulate nucleic acids, optionally mRNA encoding peptides or proteins. In an embodiment, the encapsulation percentage of mRNA by the lipid nanoparticles is at least 70%. In an embodiment, the encapsulation percentage of mRNA by the lipid nanoparticles is at least 75%. In an embodiment, the encapsulation percentage of mRNA by the lipid nanoparticles is at least 80%. In an embodiment, the encapsulation percentage of mRNA by the lipid nanoparticles is at least 85%. In an embodiment, the encapsulation percentage of mRNA by the lipid nanoparticles is at least 90%. In the examples, the encapsulation percentage of mRNA by the lipid nanoparticles is at least 95%.

[0409] In the embodiments, the composition according to any one of the foregoing embodiments is used for treatment.

[0410] In the embodiments, the composition according to any one of the foregoing embodiments is used for a method of treating or preventing a disease suitable for treatment or prevention by means of a peptide or protein encoded by mRNA, wherein the disease is optionally: (a) a protein deficiency, wherein the protein deficiency optionally affects the liver, lungs, brain or muscles; (b) an autoimmune disease; (c) an infectious disease; or (d) cancer.

[0411] In the embodiments, the composition is administered intravenously, intrathecally, or intramuscularly, or delivered via the lungs, optionally via nebulization.

[0412] Exemplary compounds

[0413] The exemplary compounds include those described in Tables 1-8.

[0414] Table 1

[0415]

[0416]

[0417]

[0418]

[0419] Table 2

[0420]

[0421]

[0422]

[0423]

[0424] Table 3

[0425]

[0426]

[0427]

[0428]

[0429] Table 4

[0430]

[0431]

[0432]

[0433]

[0434] Table 5

[0435]

[0436]

[0437]

[0438]

[0439] Table 6

[0440]

[0441]

[0442]

[0443]

[0444] Table 7

[0445]

[0446]

[0447] Table 8

[0448]

[0449] Any of the compounds identified in Tables 1 to 8 above may be provided in the form of pharmaceutically acceptable salts, and such salts are intended to be covered by this invention.

[0450] Unless otherwise stated, R = C 16 H 29 It has a structure:

[0451]

[0452] Unless otherwise stated, R = C 16 H 31 It has a structure:

[0453]

[0454] The compounds of the present invention as described herein can be prepared according to methods known in the art, including exemplary synthesis of the examples provided herein.

[0455] Nucleic acid

[0456] The compounds of the present invention, as described herein, can be used to prepare compositions that can be used to deliver nucleic acids.

[0457] Nucleic acid synthesis

[0458] Nucleic acids according to the invention can be synthesized according to any known method. For example, mRNA according to the invention can be synthesized by in vitro transcription (IVT). Briefly, IVT is typically performed using a linear or circular DNA template containing a promoter, a library of ribonucleotide triphosphates, a buffer system possibly containing DTT and magnesium ions, and a suitable RNA polymerase (e.g., T3, T7, mutant T7, or SP6 RNA polymerase), DNAse I, pyrophosphatase, and / or RNase inhibitors. The exact conditions will vary depending on the specific application.

[0459] In some embodiments, to prepare the mRNA according to the invention, a DNA template is transcribed in vitro. A suitable DNA template typically has a promoter for in vitro transcription, such as a T3, T7, mutated T7, or SP6 promoter, followed by the desired nucleotide sequence of the desired mRNA and a termination signal.

[0460] The desired mRNA sequence according to the invention can be determined using standard methods and incorporated into a DNA template. For example, virtual reverse translation is performed based on a degenerate genetic code, starting from the desired amino acid sequence (e.g., an enzyme sequence). An optimization algorithm can then be used to select suitable codons. Typically, the G / C ratio can be optimized to achieve the highest possible G / C ratio, while also considering the frequency of tRNA usage based on codon usage. The optimized RNA sequence can be constructed and displayed, for example, using a suitable display device, and compared with the original (wild-type) sequence. Secondary structures can also be analyzed to separately calculate stable and destabilized regions of the RNA.

[0461] Modified mRNA

[0462] In some embodiments, the mRNA according to the invention can be synthesized as unmodified mRNA or modified mRNA. Modified mRNA contains nucleotide modifications within the RNA. Therefore, the modified mRNA according to the invention can include nucleotide modifications, such as backbone modifications, sugar modifications, or base modifications. In some embodiments, mRNAs can be synthesized from naturally occurring nucleotides and / or nucleotide analogs (modified nucleotides), including but not limited to purines (adenine (A), guanine (G)) or pyrimidines (thymine (T), cytosine (C), uracil (U)), as modified nucleotide analogs or derivatives of purines and pyrimidines, such as 1-methyl-adenine, 2-methyl-adenine, 2-methylthio-N-6-isopentenyl-adenine, N6-methyl-adenine, N6-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), dihydrouracil, 2-thiouracil, 4-thiouracil, 5-carboxymethylaminomethyl-2-thiouracil, 5-(carboxyhydroxymethyl)uracil, 5-fluorouracil, 5-bromouracil, 5-carboxymethylaminomethyluracil, 5-methyl-2-thiouracil, 5-methyluracil, N-uracil-5-oxyacetic acid methyl ester, 5-methylaminomethyluracil 5-Methoxyaminomethyl-2-thio-uracil, 5'-methoxycarbonylmethyl-uracil, 5-methoxy-uracil, methyl uracil-5-oxyacetate, uracil-5-oxyacetate (v), 1-methyl-pseudouracil, piracetamidine, β-D-mannosyl-piracetamidine, huaitoxy glycoside, as well as aminophosphates, thiophosphates, peptide nucleotides, methylphosphonates, 7-denitroguanosine, 5-methylcytosine, and inosine. The preparation of such analogues is known to those skilled in the art, for example, according to U.S. Patent Nos. 4,373,071, 4,401,796, 4,415,732, 4,458,066, 4,500,707, 4,668,777, 4,973,679, 5,047,524, 5,132,418, 5,153,319, 5,262,530, and 5,700,642, the disclosures of which are incorporated herein by reference in their entirety.

[0463] Pharmaceutical formulations of cationic lipids and nucleic acids

[0464] In some embodiments, compounds of the present invention as described herein, as well as pharmaceutical and liposome compositions comprising such lipids, can be used in formulations to facilitate the delivery of encapsulating materials (e.g., one or more polynucleotides, such as mRNA) to one or more target cells and subsequently transfect them. For example, in some embodiments, the cationic lipids described herein (and compositions comprising such lipids, such as liposome compositions) are characterized by one or more of the following: receptor-mediated endocytosis, clathrin-mediated and pitot-mediated endocytosis, phagocytosis and macropinocytosis, fusion-promoting properties, endosome or lysosomal disruption, and / or releasability, which provide such compounds with advantages over other similarly classified lipids.

[0465] According to the present invention, nucleic acids as described herein, such as mRNA encoding proteins (e.g., full-length, fragment, or portion of a protein), can be delivered by delivery media comprising compounds of the present invention as described herein.

[0466] As used herein, the terms “delivery medium,” “transfer medium,” “nanoparticle,” or their grammatical equivalents are used interchangeably.

[0467] For example, the present invention provides compositions comprising the compounds described herein and one or more polynucleotides (e.g., pharmaceutical compositions). The compositions (e.g., pharmaceutical compositions) may also comprise one or more cationic lipids, one or more non-cationic lipids, one or more cholesterol-based lipids, and / or one or more PEG-modified lipids.

[0468] In some embodiments, the compositions exhibit enhanced (e.g., improved) transfection ability of one or more target cells. Therefore, methods for transfecting one or more target cells are also provided herein. Such methods generally involve contacting one or more target cells with the cationic lipid and / or pharmaceutical compositions disclosed herein (e.g., liposome formulations comprising one or more polynucleotides encapsulated therein), such that one or more target cells are transfected with the encapsulated material (e.g., one or more polynucleotides). As used herein, the term “transfection” refers to the introduction of one or more encapsulating materials (e.g., nucleic acids and / or polynucleotides) into cells, or preferably into target cells. The introduced polynucleotides may be stably or transiently retained in the target cells. The term “transfection efficiency” refers to the relative amount of such encapsulating material (e.g., polynucleotides) taken up, introduced, and / or expressed by the transfected target cells. In practice, transfection efficiency can be estimated by the amount of reporter polynucleotide products produced by the target cells after transfection. In some embodiments, the compounds and pharmaceutical compositions described herein exhibit high transfection efficiency, thereby increasing the likelihood that an appropriate dose of encapsulating material (e.g., one or more polynucleotides) will be delivered to the pathological site and subsequently expressed, while minimizing potential systemic side effects or toxicities associated with the compound or its encapsulated contents.

[0469] Following transfection of one or more target cells with polynucleotides, for example, encapsulated in one or more lipid nanoparticles constituting the pharmaceutical or liposome compositions disclosed herein, the production of products (e.g., peptides or proteins) encoded by such polynucleotides can preferably be stimulated, and the ability of such target cells to express polynucleotides and produce, for example, peptides or proteins of interest can be enhanced. For example, transfection of target cells with one or more compounds or pharmaceutical compositions encapsulating mRNA will enhance (i.e. increase) the production of proteins or enzymes encoded by such mRNAs.

[0470] Additionally, the delivery media described herein (e.g., liposome delivery media) can be prepared to preferentially distribute to other target tissues, cells, or organs, such as the heart, lungs, kidneys, and spleen. In embodiments, lipid nanoparticles of the present invention can be prepared to achieve enhanced delivery to target cells and tissues. For example, polynucleotides (e.g., mRNA) encapsulated in one or more compounds or pharmaceutical compositions and liposome compositions described herein can be delivered to and / or transfected into target cells or tissues. In some embodiments, the encapsulated polynucleotides (e.g., mRNA) are capable of being expressed by target cells and produced by (and in some cases secreted by) functional polypeptide products, thereby conferring, for example, beneficial properties to the target cells or tissues. Such encapsulated polynucleotides (e.g., mRNA) may encode, for example, hormones, enzymes, receptors, polypeptides, peptides, or other proteins of interest.

[0471] Liposome delivery mediators

[0472] In some embodiments, the composition is a suitable delivery medium. In examples, the composition is a liposome delivery medium, such as lipid nanoparticles.

[0473] The terms “liposome delivery medium” and “liposome composition” are used interchangeably.

[0474] Using one or more of the cationic lipids disclosed herein to enrich liposome compositions can be used as a means of improving (e.g., reducing) toxicity or otherwise imparting one or more desired properties to such enriched liposome compositions (e.g., improving the delivery of encapsulated polynucleotides to one or more target cells and / or reducing the in vivo toxicity of the liposome composition). Therefore, pharmaceutical compositions comprising one or more of the cationic lipids disclosed herein, particularly liposome compositions, are also contemplated.

[0475] Therefore, in some embodiments, the compounds of the present invention as described herein can be used as components of liposome compositions to facilitate or enhance the delivery and release of encapsulating materials (e.g., one or more therapeutic agents) to one or more target cells (e.g., by penetrating or fusing with the lipid membrane of such target cells).

[0476] As used herein, liposome delivery mediators (e.g., lipid nanoparticles) are typically characterized as microscopic vesicles with an internal water space isolated from an external medium by one or more bilayer membranes. The bilayer membranes of liposomes are typically formed from amphiphilic molecules, such as synthetic or naturally derived lipids comprising spatially separated hydrophilic and hydrophobic domains (Lasic, Trends Biotechnol., 16:307-321, 1998). The bilayer membranes of liposomes can also be formed from amphiphilic polymers and surfactants (e.g., polymeric vesicles, liposomes, etc.). In the context of this invention, liposome delivery mediators are generally used to transport desired mRNA to target cells or tissues.

[0477] In some embodiments, such compositions (e.g., liposome compositions) are loaded or otherwise encapsulated with materials such as one or more bioactive polynucleotides (e.g., mRNA).

[0478] In embodiments, the composition (e.g., a pharmaceutical composition) comprises mRNA encoding a protein encapsulated in liposomes. In embodiments, the liposomes comprise one or more cationic lipids, one or more non-cationic lipids, one or more cholesterol-based lipids, and one or more PEG-modified lipids, wherein at least one cationic lipid is a compound of the invention as described herein. In embodiments, the composition comprises mRNA encoding a protein (e.g., any protein described herein). In embodiments, the composition comprises mRNA encoding a cystic fibrosis transmembrane conduction regulator (CFTR) protein. In embodiments, the composition comprises mRNA encoding an ornithine transcarbamylase (OTC) protein.

[0479] In the embodiments, the composition (e.g., a pharmaceutical composition) comprises nucleic acid encapsulated in liposomes, wherein the liposomes contain the compounds described herein.

[0480] In these embodiments, the nucleic acid is mRNA encoding a peptide or protein. In these embodiments, the mRNA encodes a peptide or protein for delivery to or treatment of the lungs or lung cells of a subject (e.g., mRNA encoding a cystic fibrosis transmembrane conductance modulator (CFTR) protein). In these embodiments, the mRNA encodes a peptide or protein for delivery to or treatment of the liver or hepatocytes of a subject (e.g., mRNA encoding an ornithine transcarbamate (OTC) protein). Other exemplary mRNAs are also described herein.

[0481] In the embodiments, the liposome delivery medium (e.g., lipid nanoparticles) may have a net positive charge.

[0482] In the embodiments, the liposome delivery medium (e.g., lipid nanoparticles) may have a net negative charge.

[0483] In the embodiments, the liposome delivery medium (e.g., lipid nanoparticles) may have a net neutral charge.

[0484] In the embodiments, the lipid nanoparticles encapsulating nucleic acids (e.g., mRNA encoding peptides or proteins) comprise one or more compounds of the present invention as described herein.

[0485] For example, the amount of the compounds of the present invention as described herein in a composition can be described as a percentage (“wt%”) of the combined dry weight of all lipids in the composition (e.g., the combined dry weight of all lipids present in a liposome composition).

[0486] In the embodiments of the pharmaceutical compositions described herein, the compounds of the invention as described herein are present in an amount of 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 the composition (e.g., a liposome composition).

[0487] In the embodiments, the compounds of the present invention as described herein are present in amounts of 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 the composition (e.g., a liposome composition). In the embodiments, the compounds of the present invention as described herein are present in amounts of 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 the composition (e.g., a liposome delivery medium).

[0488] In the embodiments, the compounds of the present invention as described herein are present in amounts of 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 the composition (e.g., a liposome composition).

[0489] In the embodiments, the compounds of the present invention as described herein are present in amounts not exceeding 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 the composition (e.g., a liposome composition).

[0490] In examples, the composition (e.g., a liposome delivery medium, such as lipid nanoparticles) comprises about 0.1 wt% to about 20 wt% (e.g., about 0.1 wt% to about 15 wt%) of the compounds described herein. In examples, the delivery medium (e.g., a liposome delivery medium, such as lipid nanoparticles) comprises about 0.5 wt%, about 1 wt%, about 3 wt%, about 5 wt%, or about 10 wt% of the compounds described herein. In examples, the delivery medium (e.g., a liposome delivery medium, such as lipid nanoparticles) 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 the compounds described herein. In examples, this percentage results in improved beneficial effects (e.g., improved delivery to target tissues, such as the liver or lungs).

[0491] The amount of the compounds of the present invention as described herein in the composition can also be described as a percentage (“mol%”) of the combined molar amount of the total lipids in the composition (e.g., the combined molar amount of all lipids present in a liposome delivery medium).

[0492] In the embodiments of the pharmaceutical compositions described herein, the compounds of the invention as described herein are present in an amount of about 0.5 mol% to about 50 mol% (e.g., about 0.5 mol% to about 20 mol%) of the combined molar amount of all lipids present in the composition (e.g., a liposome delivery medium).

[0493] In the embodiments, the compounds of the present invention as described herein are present in amounts of 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%. In the embodiments, the compounds of the present invention as described herein are present in amounts of 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%.

[0494] In some embodiments, the compounds of the present invention as described herein may comprise from about 0.1 mol% to about 50 mol% of the total lipids in the composition (e.g., a liposome delivery medium), or from about 0.5 mol% to about 50 mol%, or from about 1 mol% to about 25 mol%, or from about 1 mol% to about 10 mol%.

[0495] In some embodiments, the compounds of the present invention as described herein may constitute a greater than about 0.1 mol%, or greater than about 0.5 mol%, or greater than about 1 mol%, or greater than about 5 mol%, or greater than about 10 mol%, or greater than about 20 mol%, or greater than about 30 mol%, or greater than about 40 mol% of the total lipids in the lipid nanoparticles.

[0496] In some embodiments, the compound as described may constitute 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%, or 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 lipids in the composition (e.g., a liposome delivery medium).

[0497] In the embodiments, the compounds of the present invention as described herein are present in amounts of 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 amount of total lipids in the composition (e.g., a liposome composition).

[0498] In the embodiments, the amounts of the compounds of the present invention as described herein are present in the combined molar amount of total lipids in the composition (e.g., a liposome composition) not exceeding 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%.

[0499] In the embodiments, this percentage leads to improved beneficial effects (e.g., improved delivery to target tissues such as the liver or lungs).

[0500] In a typical embodiment, the compositions of the present invention (e.g., liposome compositions) comprise one or more cationic lipids, one or more non-cationic lipids, one or more cholesterol-based lipids, and one or more PEG-modified lipids, wherein at least one cationic lipid is a compound of the present invention as described herein. For example, compositions suitable for practicing the present invention have four lipid components, including the compounds of the present invention described herein as cationic lipid components, non-cationic lipids, cholesterol-based lipids, and PEG-modified lipids. Non-cationic lipids may be DOPE or DEPE. Cholesterol-based lipids may be cholesterol. PEG-modified lipids may be DMG-PEG2K.

[0501] In the embodiments, the composition of the present invention includes the cationic lipid of the present invention, DMG-PEG2000, cholesterol and DOPE, and the molar ratio of cationic lipid:DMG-PEG2000:cholesterol:DOPE is 40:5:25:30.

[0502] In another embodiment, the pharmaceutical (e.g., liposome) composition comprises one or more of PEG-modified lipids, non-cationic lipids, and cholesterol lipids. In other embodiments, such pharmaceutical (e.g., liposome) compositions comprise: one or more PEG-modified lipids; one or more non-cationic lipids; and one or more cholesterol lipids. In yet another embodiment, such pharmaceutical (e.g., liposome) compositions comprise: one or more PEG-modified lipids and one or more cholesterol lipids.

[0503] In the embodiments, the composition (e.g., lipid nanoparticles) encapsulating nucleic acids (e.g., mRNA encoding peptides or proteins) includes one or more compounds of the present invention described herein and one or more lipids selected from the group consisting of cationic lipids, non-cationic lipids and polyethylene glycol-modified lipids.

[0504] In the embodiments, compositions encapsulating nucleic acids (e.g., mRNA encoding peptides or proteins) (e.g., lipid nanoparticles) include one or more compounds of the present invention as described herein, one or more lipids selected from the group consisting of cationic lipids, non-cationic lipids, and polyethylene glycol-modified lipids; and further include cholesterol-based lipids. Typically, such compositions have four lipid components, including compounds of the present invention as described herein as cationic lipid components, non-cationic lipids (e.g., DOPE), cholesterol-based lipids (e.g., cholesterol), and PEG-modified lipids (e.g., DMG-PEG2K).

[0505] In the embodiments, the composition encapsulating nucleic acids (e.g., mRNA encoding peptides or proteins) (e.g., lipid nanoparticles) includes one or more compounds of the present invention as described herein, and one or more lipids selected from the group consisting of cationic lipids, non-cationic lipids, polyethylene glycol-modified lipids, and cholesterol-based lipids.

[0506] According to various embodiments, the selection of cationic lipids, non-cationic lipids, and / or PEG-modified lipids comprising lipid nanoparticles, and the relative molar ratios of these lipids to each other, are based on the characteristics of the selected lipids, the nature of the intended target cells, and the characteristics of the mRNA to be delivered. Other considerations include, for example, the saturation of the alkyl chain and the size, charge, pH, pKa, fusion, and toxicity of the selected lipids. Therefore, the molar ratios can be adjusted accordingly.

[0507] In some embodiments, the ratios of cationic lipids to non-cationic lipids to cholesterol-based lipids to PEG-modified lipids may be between approximately 30-60:20-40:20-30:1-10. In some embodiments, the ratios of one or more cationic lipids to one or more non-cationic lipids to one or more cholesterol-based lipids to one or more PEG-modified lipids are approximately 40:30:20:10. In some embodiments, the ratios of one or more cationic lipids to one or more non-cationic lipids to one or more cholesterol-based lipids to one or more PEG-modified lipids are approximately 40:30:25:5. In some embodiments, the ratios of one or more cationic lipids to one or more non-cationic lipids to one or more cholesterol-based lipids to one or more PEG-modified lipids are approximately 40:32:25:3. In some embodiments, the ratios of one or more cationic lipids to one or more non-cationic lipids to one or more cholesterol-based lipids to one or more PEG-modified lipids are approximately 50:25:20:5.

[0508] cationic lipids

[0509] In addition to any of the compounds of the present invention as described herein, the composition may include one or more additional cationic lipids.

[0510] In some embodiments, liposomes may comprise one or more additional cationic lipids. As used herein, the phrase "cationic lipid" refers to any of a variety of lipid substances 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.

[0511] Suitable additional cationic lipids for use in the composition include cationic lipids as described in the literature.

[0512] assist lipids

[0513] Compositions (e.g., liposome compositions) may also include one or more accessory lipids. Such accessory lipids comprise noncationic lipids. As used herein, the phrase "noncationic lipid" refers to any neutral, zwitterionic, or anionic lipid. As used herein, the phrase "anionic lipid" refers to any of a variety of lipid substances that carry a net negative charge at a selected pH, such as physiological pH. Noncationic lipids include, but are not limited to, distearylphosphatidylcholine (DSPC), dioleoylphosphatidylcholine (DOPC), dipalmitoylphosphatidylcholine (DPPC), dioleoylphosphatidylglycerol (DOPG), dipalmitoylphosphatidylglycerol (DPPG), dioleoylphosphatidylethanolamine (DOPE), 1,2-diethylene glycol-sn-glycerol-3-phosphate ethanolamine (DEPE), palmitoyloleoylphosphatidylcholine (POPC), and palmitoyloleoylphosphatidylethanolamine (P... Dioleoylphosphatidylethanolamine (DOPE), dioleoylphosphatidylethanolamine 4-(N-maleimidemethyl)-cyclohexane-1-carboxylate (DOPE-mal), dipalmitoylphosphatidylethanolamine (DPPE), dimyristoylphosphatidylethanolamine (DMPE), distearate-phosphatidylethanolamine (DSPE), 16-O-monomethylPE, 16-O-dimethylPE, 18-1-transPE, L-stearoyl-2-oleoylphosphatidylethanolamine (SOPE), or mixtures thereof. The non-cationic or auxiliary lipid suitable for practicing the present invention is dioleoylphosphatidylethanolamine (DOPE). Alternatively, 1,2-diethylene glycol-sn-glycerol-3-phosphate ethanolamine (DEPE) can be used as a non-cationic or auxiliary lipid.

[0514] In some embodiments, the non-cationic lipid is a neutral lipid, i.e., a lipid that does not carry a net charge under the conditions of formulation and / or application of the composition.

[0515] In some embodiments, non-cationic lipids may be present in the composition at 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 the composition. In some embodiments, the total non-cationic lipids may be present in the composition at 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 the composition. In some embodiments, the percentage of non-cationic lipids in the liposomes 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 total non-cationic lipids in the liposomes 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 lipids in the liposomes may not exceed about 5 mol%, not exceed about 10 mol%, not exceed about 20 mol%, not exceed about 30 mol%, or not exceed about 40 mol%. In some embodiments, the percentage of total non-cationic lipids in the liposomes may not exceed about 5 mol%, not exceed about 10 mol%, not exceed about 20 mol%, not exceed about 30 mol%, or not exceed about 40 mol%.

[0516] In some embodiments, non-cationic lipids may be present in the composition at a weight percentage (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 the composition. In some embodiments, the percentage of non-cationic lipids in the liposomes may be greater than about 5% by weight, greater than about 10% by weight, greater than about 20% by weight, greater than about 30% by weight, or greater than about 40% by weight. In some embodiments, the percentage of total non-cationic lipids in the liposomes may be greater than about 5% by weight, greater than about 10% by weight, greater than about 20% by weight, greater than about 30% by weight, or greater than about 40% by weight. In some embodiments, the percentage of non-cationic lipids in the liposomes may not exceed about 5% by weight, not more than about 10% by weight, not more than about 20% by weight, not more than about 30% by weight, or not more than about 40% by weight. In some embodiments, the percentage of total non-cationic lipids in the liposomes may not exceed about 5% by weight, not more than about 10% by weight, not more than about 20% by weight, not more than about 30% by weight, or not more than about 40% by weight.

[0517] Cholesterol-based lipids

[0518] In some embodiments, the composition (e.g., a liposome composition) comprises one or more cholesterol-based lipids. For example, a suitable cholesterol-based lipid for practicing the present invention is cholesterol. Other suitable cholesterol-based lipids include, for example, DC-Chol (N,N-dimethyl-N-ethylcarboxyamide cholesterol), 1,4-bis(3-N-oleoaminopropyl)piperazine (Gao et al., Biochem.Biophys.Res.Comm. 179,280 (1991); Wolf et al., BioTechniques 23,139 (1997); U.S. Patent No. 5,744,335), or imidazole cholesterol ester (ICE) having the following structure:

[0519]

[0520] In some embodiments, cholesterol-based lipids may be present at a molar ratio (mol%) of about 1% to about 30% or about 5% to about 20% of the total lipids present in the liposomes. In some embodiments, the percentage of cholesterol-based lipids in the lipid nanoparticles 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 lipids in the lipid nanoparticles may not exceed about 5 mol%, not exceed about 10 mol%, not exceed about 20 mol%, not exceed about 30 mol%, or not exceed about 40 mol%.

[0521] In some embodiments, cholesterol-based lipids may be present at a weight percentage (wt%) of about 1% to about 30% or about 5% to about 20% of the total lipids present in the liposomes. In some embodiments, the percentage of cholesterol-based lipids in the lipid nanoparticles 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 lipids in the lipid nanoparticles may not exceed about 5% wt, not more than about 10% wt, not more than about 20% wt, not more than about 30% wt, or not more than about 40% wt.

[0522] PEGylated lipids

[0523] In some embodiments, the composition (e.g., a liposome composition) comprises one or more additional PEGylated lipids. A suitable PEG-modified or PEGylated lipid for practicing the present invention is 1,2-dimyristoyl-rac-glycerol-3-methoxypolyethylene glycol-2000 (DMG-PEG2K).

[0524] For example, the present invention also contemplates the use of polyethylene glycol (PEG)-modified phospholipids and derived lipids, such as derived ceramides (PEG-CER), comprising N-octanoyl-sphingosine-1-[succinyl(methoxypolyethylene glycol)-2000] (C8 PEG-2000 ceramide) with one or more compounds of the present invention described herein, and in some embodiments, in combination with other lipids including liposomes. In some embodiments, particularly useful exchangeable lipids are those having shorter acyl chains (e.g., C8 PEG-2000 ceramide). 14 Or C 18 PEG-ceramide.

[0525] The anticipated additional PEG-modified lipids (also referred to herein as polyethylene glycol-modified lipids, a term used interchangeably with PEG-modified lipids) include, but are not limited to, covalently linked to lipids having C6-C6. 20The lipids have alkyl chains of up to 5 kDa in length, consisting of polyethylene glycol chains. In some embodiments, the PEG-modified lipids or PEGylated lipids are PEGylated cholesterol or PEG-2K. The addition of such components can prevent complex aggregation and can also provide a means of increasing cycle life and increasing the delivery of lipid-nucleic acid compositions to target cells (Klibanov et al. (1990), FEBS Letters, 268(1):235-237), or the components can be selected for rapid in vivo exchange of the formulation (see U.S. Patent No. 5,885,613).

[0526] The additional PEG-modified phospholipids and derived lipids of the present invention may be present in a molar ratio (mol%) of about 0% to about 10%, about 0.5% to about 10%, about 1% to about 10%, about 2% to about 10%, or about 3% to about 5% of the total lipids present in the composition (e.g., liposome composition).

[0527] Pharmaceutical preparations and therapeutic uses

[0528] The compounds of the present invention as described herein can be used to prepare compositions (e.g., constructing liposome compositions) that facilitate or enhance the delivery and release of encapsulating materials (e.g., one or more therapeutic polynucleotides) into one or more target cells (e.g., by permeation or fusion with the lipid membrane of such target cells).

[0529] For example, when a liposome composition (e.g., lipid nanoparticles) contains or otherwise enriches one or more of the compounds disclosed herein, a phase transition in the lipid bilayer of one or more target cells can facilitate the delivery of encapsulating materials (e.g., one or more therapeutic polynucleotides encapsulated in lipid nanoparticles) to one or more target cells.

[0530] Similarly, in some embodiments, the compounds of the present invention described herein can be used to prepare liposomes characterized by their reduced in vivo toxicity. In some embodiments, the reduced toxicity is a function of the high transfection efficiency associated with the compositions disclosed herein, making it possible to administer reduced amounts of such compositions to subjects to achieve the desired therapeutic response or outcome.

[0531] Therefore, pharmaceutical formulations including the described compounds and the nucleic acids provided by this invention can be used for a variety of therapeutic purposes. To facilitate the delivery of nucleic acids in vivo, the compounds and nucleic acids described herein can be formulated in combination with one or more additional pharmaceutical carriers, targeting ligands, or stabilizers. In some embodiments, the compounds described herein can be formulated using a premixed lipid solution. In other embodiments, post-insertion techniques can be used to formulate compositions including the compounds described herein into lipid membranes of nanoparticles. Techniques for formulating and administering pharmaceuticals are available in Remington's Pharmaceutical Sciences, Mack Publishing Co., Easton, Pa., latest edition.

[0532] Suitable routes of administration include, for example, oral, rectal, vaginal, transmucosal, pulmonary (including intratracheal or inhalation), or enteral administration; parenteral delivery, including intradermal, transdermal (local), intramuscular, subcutaneous, and intramedullary injection; and intrathecal, direct intravenous, intravenous, intraperitoneal, or intranasal administration. In certain embodiments, intramuscular administration is to muscle selected from the group consisting of skeletal muscle, smooth muscle, and cardiac muscle. In some embodiments, administration results in nucleic acid delivery to muscle cells. In some embodiments, administration results in nucleic acid delivery to hepatocytes (i.e., liver cells).

[0533] The common route of administration for the liposome composition of the present invention is intravenous delivery, specifically when treating metabolic disorders, particularly those affecting the liver (e.g., ornithine transcarbamate (OTC) deficiency). Alternatively, depending on the disease or condition to be treated, the liposome composition may be administered via pulmonary delivery (e.g., for the treatment of cystic fibrosis). For vaccination, the liposome composition of the present invention is typically administered intramuscularly. Diseases or conditions affecting the eye may be treated by intravitreal administration of the liposome composition of the present invention.

[0534] Alternatively or otherwise, the pharmaceutical formulations of the present invention can be administered locally rather than systemically, for example, by direct injection into the target tissue, preferably in the form of a sustained-release formulation. Local delivery can be influenced in various ways depending on the target tissue. Exemplary tissues in which the delivered mRNA can 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. In an example, the target tissue is in the liver. For example, an aerosol containing the composition of the present invention can be inhaled (for nasal, tracheal, or bronchial delivery); the composition of the present invention can be injected into the site of injury, disease manifestation, or pain; the composition can be provided in the form of lozenges for oral, tracheal, or esophageal application; it can be supplied to the stomach or intestine in the form of a liquid, tablet, or capsule, or to the rectum or vagina in the form of a suppository; or even delivered to the eyes using creams, drops, or even injections.

[0535] The compositions described herein may include mRNA-encoded peptides, including peptides (e.g., polypeptides, such as proteins) as described herein.

[0536] In the examples, mRNA encodes a polypeptide.

[0537] In the examples, mRNA encodes proteins.

[0538] This article describes exemplary peptides (e.g., exemplary proteins encoded by mRNA) that are encoded by mRNA.

[0539] The present invention provides a method for delivering a composition having a full-length mRNA molecule encoding a peptide or protein of interest, the composition being used to treat a subject, such as a human subject or cells of a human subject, or cells of a human subject that have been treated and delivered to the human subject's cells.

[0540] Therefore, in some embodiments, the present invention provides a method for preparing a therapeutic composition comprising full-length mRNA encoding a peptide or protein, the therapeutic composition being delivered to or used to treat the lungs or lung cells of a subject. In some embodiments, the present invention provides a method for preparing a therapeutic composition having full-length mRNA encoding a cystic fibrosis transmembrane conduction regulator (CFTR) protein. In some embodiments, the present invention provides a method for preparing a therapeutic composition having full-length mRNA encoding a member of the ATP-binding box subfamily A 3 protein. In some embodiments, the present invention provides a method for preparing a therapeutic composition having full-length mRNA encoding a dynein axon intermediate chain 1 protein. In some embodiments, the present invention provides a method for preparing a therapeutic composition having full-length mRNA encoding a dynein axon heavy chain 5 (DNAH5) protein. In some embodiments, the present invention provides a method for preparing a therapeutic composition having full-length mRNA encoding an α-1-antitrypsin protein. In some embodiments, the present invention provides a method for preparing a therapeutic composition having full-length mRNA encoding a forkhead box P3 (FOXP3) protein. In some embodiments, the present invention provides a method for producing a therapeutic composition having a full-length mRNA encoding one or more surfactant proteins (e.g., surfactant protein A, surfactant protein B, surfactant protein C, and surfactant protein D).

[0541] In some embodiments, the present invention provides a method for preparing a therapeutic composition having full-length mRNA encoding a peptide or protein, the therapeutic composition being delivered to or used to treat the liver or hepatocytes of a subject. Such peptides and polypeptides may include those associated with urea cycle disorders, lysosomal storage disorders, glycogen storage disorders, amino acid metabolism disorders, lipid metabolism or fibrosis disorders, methylmalonic acidemia, or any other metabolic disorder, for which delivery of enriched full-length mRNA to or treatment of the liver or hepatocytes provides a therapeutic benefit.

[0542] In some embodiments, the present invention provides a method for preparing a therapeutic composition having full-length mRNA encoding a protein associated with urea cycle disorder. In some embodiments, the present invention provides a method for preparing a therapeutic composition having full-length mRNA encoding ornithine transcarbamate (OTC) protein. In some embodiments, the present invention provides a method for preparing a therapeutic composition having full-length mRNA encoding argininosuccinate synthase 1 protein. In some embodiments, the present invention provides a method for preparing a therapeutic composition having full-length mRNA encoding carbamoyl phosphate synthase I protein. In some embodiments, the present invention provides a method for preparing a therapeutic composition having full-length mRNA encoding argininosuccinate lyase protein. In some embodiments, the present invention provides a method for preparing a therapeutic composition having full-length mRNA encoding arginase protein.

[0543] In some embodiments, the present invention provides a method for preparing a therapeutic composition having full-length mRNA encoding a protein associated with lysosomal storage disorders. In some embodiments, the present invention provides a method for preparing a therapeutic composition having full-length mRNA encoding α-galactosidase protein. In some embodiments, the present invention provides a method for preparing a therapeutic composition having full-length mRNA encoding glucocerebrosidase protein. In some embodiments, the present invention provides a method for preparing a therapeutic composition having full-length mRNA encoding isocyanate-2-sulfatase protein. In some embodiments, the present invention provides a method for preparing a therapeutic composition having full-length mRNA encoding idurosidase protein. In some embodiments, the present invention provides a method for preparing a therapeutic composition having full-length mRNA encoding N-acetyl-α-D-glucosamine sidase protein. In some embodiments, the present invention provides a method for preparing a therapeutic composition having full-length mRNA encoding heparin N-sulfatase protein. In some embodiments, the present invention provides a method for preparing a therapeutic composition having full-length mRNA encoding galactosamine-6-sulfatase protein. In some embodiments, the present invention provides a method for preparing a therapeutic composition having full-length mRNA encoding β-galactosidase protein. In some embodiments, the present invention provides a method for preparing a therapeutic composition having full-length mRNA encoding lysosomal lipase protein. In some embodiments, the present invention provides a method for preparing a therapeutic composition having full-length mRNA encoding arylsulfatase B (N-acetylgalactosamine-4-sulfatase) protein. In some embodiments, the present invention provides a method for preparing a therapeutic composition having full-length mRNA encoding transcription factor EB (TFEB).

[0544] In some embodiments, the present invention provides a method for preparing a therapeutic composition having full-length mRNA encoding a protein associated with glycogen storage impairment. In some embodiments, the present invention provides a method for preparing a therapeutic composition having full-length mRNA encoding an acidic α-glucosidase protein. In some embodiments, the present invention provides a method for preparing a therapeutic composition having full-length mRNA encoding a glucose-6-phosphatase (G6PC) protein. In some embodiments, the present invention provides a method for preparing a therapeutic composition having full-length mRNA encoding a liver glycogen phosphorylase protein. In some embodiments, the present invention provides a method for preparing a therapeutic composition having full-length mRNA encoding a muscle phosphoglycerate mutant enzyme protein. In some embodiments, the present invention provides a method for preparing a therapeutic composition having full-length mRNA encoding a glycogen debranching enzyme.

[0545] In some embodiments, the present invention provides a method for preparing a therapeutic composition having full-length mRNA encoding a protein related to amino acid metabolism. In some embodiments, the present invention provides a method for preparing a therapeutic composition having full-length mRNA encoding phenylalanine hydroxylase. In some embodiments, the present invention provides a method for preparing a therapeutic composition having full-length mRNA encoding glutaryl-CoA dehydrogenase. In some embodiments, the present invention provides a method for preparing a therapeutic composition having full-length mRNA encoding propionyl-CoA carboxylase. In some embodiments, the present invention provides a method for preparing a therapeutic composition having full-length mRNA encoding oxalate alanine-glyoxylate aminotransferase.

[0546] In some embodiments, the present invention provides a method for preparing a therapeutic composition having full-length mRNA encoding a protein associated with lipid metabolism or fibrosis disorders. In some embodiments, the present invention provides a method for preparing a therapeutic composition having full-length mRNA encoding an mTOR inhibitor. In some embodiments, the present invention provides a method for preparing a therapeutic composition having full-length mRNA encoding the ATPase phospholipid transporter 8B1 (ATP8B1) protein. In some embodiments, the present invention provides a method for preparing a therapeutic composition having full-length mRNA encoding one or more NF-κB inhibitors, such as I-κBα, interferon-associated developmental regulator 1 (IFRD1), and Sirtuin 1 (SIRT1). In some embodiments, the present invention provides a method for preparing a therapeutic composition having full-length mRNA encoding a PPAR-γ protein or an active variant.

[0547] In some embodiments, the present invention provides a method for preparing a therapeutic composition having full-length mRNA encoding a protein associated with methylmalonic acidemia. For example, in some embodiments, the present invention provides a method for preparing a therapeutic composition having full-length mRNA encoding a methylmalonyl-CoA mutant enzyme protein. In some embodiments, the present invention provides a method for preparing a therapeutic composition having full-length mRNA encoding a methylmalonyl-CoA epimerase protein.

[0548] In some embodiments, the present invention provides a method for preparing a therapeutic composition having full-length mRNA, the delivery of which to or treatment of the liver may provide a therapeutic benefit. In some embodiments, the present invention provides a method for preparing a therapeutic composition having full-length mRNA encoding ATP7B protein (also known as Wilson's disease protein). In some embodiments, the present invention provides a method for preparing a therapeutic composition having full-length mRNA encoding bilirubinogen deaminase. In some embodiments, the present invention provides a method for preparing a therapeutic composition having full-length mRNA encoding one or more coagulating enzymes, such as factors VIII, IX, VII, and X. In some embodiments, the present invention provides a method for preparing a therapeutic composition having full-length mRNA encoding human hemochromatosis (HFE) protein.

[0549] In some embodiments, the present invention provides a method for preparing a therapeutic composition having full-length mRNA encoding a peptide or protein, the therapeutic composition being delivered to or treating the cardiovascular system or cardiovascular cells of a subject. In some embodiments, the present invention provides a method for preparing a therapeutic composition having full-length mRNA encoding vascular endothelial growth factor A protein. In some embodiments, the present invention provides a method for preparing a therapeutic composition having full-length mRNA encoding relaxin protein. In some embodiments, the present invention provides a method for preparing a therapeutic composition having full-length mRNA encoding bone morphogenetic protein 9 protein. In some embodiments, the present invention provides a method for preparing a therapeutic composition having full-length mRNA encoding bone morphogenetic protein 2 receptor protein.

[0550] In some embodiments, the present invention provides a method for preparing a therapeutic composition having full-length mRNA encoding a peptide or protein, the therapeutic composition being delivered to or treating muscle or muscle cells of a subject. In some embodiments, the present invention provides a method for preparing a therapeutic composition having full-length mRNA encoding a dystrophin. In some embodiments, the present invention provides a method for preparing a therapeutic composition having full-length mRNA encoding a human mitochondrial protein (frataxin). In some embodiments, the present invention provides a method for preparing a therapeutic composition having full-length mRNA encoding a peptide or protein, the therapeutic composition being delivered to or treating cardiac muscle or cardiomyocytes of a subject. In some embodiments, the present invention provides a method for preparing a therapeutic composition having full-length mRNA encoding a protein regulating one or both of potassium and sodium channels in muscle tissue or muscle cells. In some embodiments, the present invention provides a method for preparing a therapeutic composition having full-length mRNA encoding a protein regulating the Kv7.1 channel in muscle tissue or muscle cells. In some embodiments, the present invention provides a method for preparing a therapeutic composition having full-length mRNA encoding a protein regulating the Nav1.5 channel in muscle tissue or muscle cells.

[0551] In some embodiments, the present invention provides a method for preparing a therapeutic composition having full-length mRNA encoding a peptide or protein, the therapeutic composition being delivered to or treating the nervous system or nervous system cells of a subject. For example, in some embodiments, the present invention provides a method for preparing a therapeutic composition having full-length mRNA encoding a viable motor neuron 1 protein. For example, in some embodiments, the present invention provides a method for preparing a therapeutic composition having full-length mRNA encoding a viable motor neuron 2 protein. In some embodiments, the present invention provides a method for preparing a therapeutic composition having full-length mRNA encoding a human mitochondrial protein (frataxin). In some embodiments, the present invention provides a method for preparing a therapeutic composition having full-length mRNA encoding an ATP-binding box subfamily D member 1 (ABCD1) protein. In some embodiments, the present invention provides a method for preparing a therapeutic composition having full-length mRNA encoding a CLN3 protein.

[0552] In some embodiments, the present invention provides a method for preparing a therapeutic composition having full-length mRNA encoding a peptide or protein, the therapeutic composition being delivered to or treating the blood or bone marrow or blood cells or bone marrow cells of a subject. In some embodiments, the present invention provides a method for preparing a therapeutic composition having full-length mRNA encoding a β-globulin. In some embodiments, the present invention provides a method for preparing a therapeutic composition having full-length mRNA encoding a Bruton's tyrosine kinase protein. In some embodiments, the present invention provides a method for preparing a therapeutic composition having full-length mRNA encoding one or more coagulating enzymes, such as factor VIII, factor IX, factor VII, and factor X.

[0553] In some embodiments, the present invention provides a method for preparing a therapeutic composition having full-length mRNA encoding a peptide or protein, the therapeutic composition being delivered to or used to treat the kidney or kidney cells of a subject. In some embodiments, the present invention provides a method for preparing a therapeutic composition having full-length mRNA encoding a type IV collagen α5 chain (COL4A5) protein.

[0554] In some embodiments, the present invention provides a method for preparing a therapeutic composition having full-length mRNA encoding a peptide or protein, the therapeutic composition being delivered to or treating the eye or ocular cells of a subject. In some embodiments, the present invention provides a method for preparing a therapeutic composition having full-length mRNA encoding an ATP-binding box subfamily A member 4 (ABCA4) protein. In some embodiments, the present invention provides a method for preparing a therapeutic composition having full-length mRNA encoding a retinaldehyde chitin protein. In some embodiments, the present invention provides a method for preparing a therapeutic composition having full-length mRNA encoding a retinal pigment epithelium-specific 65 kDa (RPE65) protein. In some embodiments, the present invention provides a method for preparing a therapeutic composition having full-length mRNA encoding a 290 kDa centsosome protein (CEP290).

[0555] In some embodiments, the present invention provides a method for preparing a therapeutic composition having full-length mRNA encoding a peptide or protein, the therapeutic composition being used for delivering a vaccine to a subject or the subject's cells, or for treating with a vaccine. For example, in some embodiments, the present invention provides a method for preparing a therapeutic composition having full-length mRNA encoding an antigen from an infectious source such as a virus. In some embodiments, the present invention provides a method for preparing a therapeutic composition having full-length mRNA encoding an antigen from an influenza virus. In some embodiments, the present invention provides a method for producing a therapeutic composition having full-length mRNA encoding an antigen from a respiratory syncytial virus. In some embodiments, the present invention provides a method for preparing a therapeutic composition having full-length mRNA encoding an antigen from a rabies virus. In some embodiments, the present invention provides a method for preparing a therapeutic composition having full-length mRNA encoding an antigen from a cytomegalovirus. In some embodiments, the present invention provides a method for producing a therapeutic composition having full-length mRNA encoding an antigen from a rotavirus. In some embodiments, the present invention provides a method for preparing a therapeutic composition having full-length mRNA encoding an antigen from a hepatitis virus, such as hepatitis A virus, hepatitis B virus, or hepatitis C virus. In some embodiments, the present invention provides a method for preparing a therapeutic composition having full-length mRNA encoding an antigen from a human papillomavirus. In some embodiments, the present invention provides a method for preparing a therapeutic composition having full-length mRNA encoding an antigen from a herpes simplex virus, such as herpes simplex virus 1 or herpes simplex virus 2. In some embodiments, the present invention provides a method for preparing a therapeutic composition having full-length mRNA encoding an antigen from a human immunodeficiency virus, such as human immunodeficiency virus type 1 or human immunodeficiency virus type 2. In some embodiments, the present invention provides a method for preparing a therapeutic composition having full-length mRNA encoding an antigen from a human pneumonia virus. In some embodiments, the present invention provides a method for preparing a therapeutic composition having full-length mRNA encoding an antigen from a human parainfluenza virus, such as human parainfluenza virus type 1, human parainfluenza virus type 2, or human parainfluenza virus type 3. In some embodiments, the present invention provides a method for preparing a therapeutic composition having full-length mRNA encoding an antigen from a malaria virus. In some embodiments, the present invention provides a method for preparing a therapeutic composition having full-length mRNA encoding an antigen from a Zika virus. In some embodiments, the present invention provides a method for preparing a therapeutic composition having full-length mRNA encoding an antigen from a Chikungunya virus.

[0556] In some embodiments, the present invention provides a method for preparing a therapeutic composition having full-length mRNA encoding an antigen associated with a subject's cancer or an antigen identified from the subject's cancer cells. In some embodiments, the present invention provides a method for preparing a therapeutic composition having full-length mRNA encoding an antigen determined from the subject's own cancer cells, i.e., providing a personalized cancer vaccine. In some embodiments, the present invention provides a method for preparing a therapeutic composition having full-length mRNA encoding an antigen expressed from a mutated KRAS gene.

[0557] In some embodiments, the present invention provides a method for preparing a therapeutic composition having a full-length mRNA encoding an antibody. In some embodiments, the antibody may be a bispecific antibody. In some embodiments, the antibody may be part of a fusion protein. In some embodiments, the present invention provides a method for preparing a therapeutic composition having a full-length mRNA encoding an antibody against OX40. In some embodiments, the present invention provides a method for preparing a therapeutic composition having a full-length mRNA encoding an antibody against VEGF. In some embodiments, the present invention provides a method for preparing a therapeutic composition having a full-length mRNA encoding an antibody against tissue necrosis factor α. In some embodiments, the present invention provides a method for preparing a therapeutic composition having a full-length mRNA encoding an antibody against CD3. In some embodiments, the present invention provides a method for preparing a therapeutic composition having a full-length mRNA encoding an antibody against CD19.

[0558] In some embodiments, the present invention provides a method for preparing a therapeutic composition having a full-length mRNA encoding an immunomodulatory agent. In some embodiments, the present invention provides a method for preparing a therapeutic composition having a full-length mRNA encoding interleukin-12. In some embodiments, the present invention provides a method for preparing a therapeutic composition having a full-length mRNA encoding interleukin-23. In some embodiments, the present invention provides a method for preparing a therapeutic composition having a full-length mRNA encoding a constitutively active variant of one or more interferon gene (STING) protein stimulators.

[0559] In some embodiments, the present invention provides a method for preparing a therapeutic composition having a full-length mRNA encoding a nuclease. In some embodiments, the present invention provides a method for preparing a therapeutic composition having a full-length mRNA encoding an RNA-directed DNA nuclease protein, such as Cas9 protein. In some embodiments, the present invention provides a method for preparing a therapeutic composition having a full-length mRNA encoding a wide range of nuclease proteins. In some embodiments, the present invention provides a method for preparing a therapeutic composition having a full-length mRNA encoding a transcription activator-like effector nuclease protein. In some embodiments, the present invention provides a method for preparing a therapeutic composition having a full-length mRNA encoding a zinc finger nuclease protein.

[0560] delivery method

[0561] The delivery routes used in the methods of this invention allow for non-invasive self-administration of the compounds of this invention. In some embodiments, the methods involve intratracheal or pulmonary administration of a composition comprising mRNA encoding a therapeutic protein in a suitable transfection or lipocarrier medium, as described above, via nebulization, vaporization, or infusion. In some embodiments, the protein is encapsulated in liposomes. In some embodiments, the liposomes comprise lipids which are compounds of this invention. As used below, administration of the compounds of this invention comprises administration of a composition comprising the compounds of this invention.

[0562] Although local lung cells and tissues represent potential targets for biological reservoirs or repositories of mRNA-encoded proteins, the applicant has found that application of the compounds of the present invention to the lungs via nebulization, vaporization, or infusion results in the distribution of even non-secreting proteins outside lung cells. Without wishing to be bound by any particular theory, it is contemplated that the nanoparticle compositions of the present invention cross the lung airway-blood barrier to achieve intact nanoparticle transfer to non-lung cells and tissues, such as the heart, liver, and spleen, resulting in the production of encoded proteins in these non-lung tissues. Therefore, the uses of the compounds and methods of the present invention extend beyond the production of therapeutic proteins in lung cells and lung tissues and can be used for delivery to non-lung target cells and / or tissues. They can be used to manage and treat a wide range of diseases, and specifically peripheral diseases caused by deficiencies in secretory and non-secreting proteins and / or enzymes (e.g., one or more lysosomal storage disorders). In some embodiments, the compounds of the present invention used in the methods of the present invention achieve the distribution of mRNA-encapsulated nanoparticles in the liver, spleen, heart, and / or other non-lung cells and the production of encoded proteins in the liver, spleen, heart, and / or other non-lung cells. For example, administering the compounds of the present invention to the lungs via nebulization, infusion, or drip will result in the composition itself and its protein products (e.g., functional β-galactosidase protein) being detectable in local lung cells and tissues, as well as peripheral target cells, tissues, and organs, due to the translocation of mRNA and delivery medium to non-lung cells.

[0563] In some embodiments, the compounds of the present invention can be used in the methods of the present invention to specifically target peripheral cells or tissues. Following pulmonary delivery, it is envisioned that the compounds of the present invention cross the pulmonary airway-blood barrier and distribute to cells other than local lung cells. Therefore, the compounds disclosed herein can be administered to a subject via the pulmonary administration route using a variety of methods known to those skilled in the art (e.g., by inhalation) and distributed to local target cells and tissues of the lungs, as well as to cells in peripheral non-pulmonary cells and tissues (e.g., liver, spleen, kidney, heart, skeletal muscle, lymph nodes, brain, cerebrospinal fluid, and plasma). Thus, both local lung cells and peripheral non-pulmonary cells can act as biological reservoirs or repositories capable of producing and / or secreting translational products encoded by one or more polynucleotides. Therefore, the present invention is not limited to the treatment of lung diseases or symptoms, but can be used as a non-invasive means of promoting the delivery of polynucleotides or the production of enzymes and proteins encoded by them in peripheral organs, tissues, and cells (e.g., hepatocytes) that would otherwise only be achievable through systemic administration. Exemplary peripheral non-lung cells include, but are not limited to, hepatocytes, epithelial cells, hematopoietic cells, epithelial cells, endothelial cells, osteocytes, stem cells, mesenchymal cells, nerve cells, cardiomyocytes, lipid cells, vascular smooth muscle cells, cardiomyocytes, skeletal muscle cells, β cells, pituitary cells, synovial endothelial cells, ovarian cells, testicular cells, fibroblasts, B cells, T cells, reticulocytes, leukocytes, granulocytes, and tumor cells.

[0564] Following administration of the composition to a subject, and for at least approximately one to seven days or longer after administration of the compound to the subject, a protein product (e.g., a functional protein or enzyme) encoded by mRNA can be detected in peripheral target tissues. The amount of protein product required to achieve a therapeutic effect will vary depending on the condition being treated, the encoded protein, and the patient's condition. For example, for at least approximately 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 after administration of the compound to the subject, the amount may be at least 0.025–1.5 μg / ml (e.g., at least 0.050 μg / ml, at least 0.075 μg / ml, at least 0.1 μg / ml, at least 0. Protein products were detected in peripheral target tissues at concentrations (e.g., therapeutic concentrations) of at least 2 μg / ml, at least 0.3 μg / ml, at least 0.4 μg / ml, at least 0.5 μg / ml, at least 0.6 μg / ml, at least 0.7 μg / ml, at least 0.8 μg / ml, at least 0.9 μg / ml, at least 1.0 μg / ml, at least 1.1 μg / ml, at least 1.2 μg / ml, at least 1.3 μg / ml, at least 1.4 μg / ml, or at least 1.5 μg / ml.

[0565] It has been demonstrated that nucleic acids can be delivered to the lungs by intratracheal administration of a liquid suspension of the compound and by inhalation of an aerosol mist generated by a liquid nebulizer or by using a dry powder device (such as the device described in U.S. Patent 5,780,014, which is incorporated herein by reference).

[0566] In some embodiments, the compounds of the invention can be formulated such that they can be atomized or otherwise delivered as particulate liquids or solids before or after administration to a subject. Such compounds can be administered with the aid of one or more suitable devices for administering such solid or liquid particulate compositions (e.g., atomized aqueous solutions or suspensions) to produce particles readily breathable or inhaled by the subject. In some embodiments, such devices (e.g., metered-dose inhalers, jet nebulizers, ultrasonic nebulizers, dry powder inhalers, propellant-based inhalers, or blowpipes) facilitate the administration of a predetermined mass, volume, or dose of the composition (e.g., about 0.5 mg / kg of mRNA per dose). For example, in some 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 some embodiments, the compounds of the invention can be formulated as particulate powders for inhalation (e.g., inhalable dry particles). In some embodiments, the compositions of the invention formulated into inhalable particles have an appropriate size such that they can be inhaled by a subject or delivered using a suitable device (e.g., with an average D50 or D90 particle size of less than about 500 μm, 400 μm, 300 μm, 250 μm, 200 μm, 150 μm, 100 μm, 75 μm, 50 μm, 25 μm, 20 μm, 15 μm, 12.5 μm, 10 μm, 5 μm, 2.5 μm or smaller). In yet other embodiments, the compounds of the invention are formulated to contain one or more pulmonary surfactants (e.g., lamellar bodies). In some embodiments, the compound of the invention is administered to a subject at 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, or 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.In some embodiments, the compound of the invention is 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, at least 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 of mRNA is administered in one or more doses.

[0567] Example

[0568] Although certain compounds, compositions, and methods of the present invention have been specifically described with reference to some embodiments, the following embodiments are for illustrative purposes only and are not intended to limit the scope of the invention.

[0569] Synthesis scheme of vanillic acid lipid

[0570]

[0571] Synthesis of 3-(dimethylamino)propyl 4-hydroxy-3-methoxybenzoate (3)

[0572]

[0573] Oxaloyl chloride (2.0 mL, 23.8 mmol) was added to a suspension of vanillic acid 1 (1.0 g, 5.9 mmol) in 25 mL of dichloromethane at 0 °C, followed by the addition of dimethylformamide (1 drop), and the resulting mixture was stirred at this temperature for 2 hours. The reaction mixture was evaporated to dryness, and the residue was dissolved in 20 mL of dichloromethane. After cooling to 0 °C, 3-(dimethylamino)prop-1-ol 2 (0.7 mL, 5.9 mmol) was slowly added, and the reaction mixture was stirred overnight at room temperature. The precipitate was filtered to yield 3-(dimethylamino)propyl 4-hydroxy-3-methoxybenzoic acid 3 (1.18 g, 79%) as a white solid.

[0574] Synthesis of 4-((4-(bis(2-((tert-butyldimethylsilyl)oxy)dodecyl)amino)butyryl)oxy)-3-methoxybenzoic acid 3-(dimethylamino)propyl ester (4)

[0575]

[0576] Oxaloyl chloride (0.15 mL, 1.72 mmol) was added to a solution of 4-(bis(2-((tert-butyldimethylsilyl)oxy)dodecyl)amino)butyric acid AIM-3-E12 (1.0 g, 1.43 mmol) in 20 mL of dichloromethane at 0 °C, followed by the addition of dimethylformamide (1 drop), and the mixture was stirred at 0 °C for 2 hours. The reaction mixture was evaporated to dryness, and the residue was dissolved in 20 mL of dichloromethane. After cooling to 0 °C, 3-(dimethylamino)propyl 4-hydroxy-3-methoxybenzoate (0.18 g, 0.7 mmol) was added, followed by the addition of pyridine (0.4 mL, 4.9 mmol), and the reaction mixture was stirred overnight at room temperature. Ice was added to quench the reaction, and the organic layer was washed with brine and dried over anhydrous sodium sulfate. After filtration and concentration, the crude product was purified by rapid chromatography to obtain 4-((4-(bis(2-((tert-butyldimethylsilyl)oxy)dodecyl)amino)butyryl)oxy)-3-methoxybenzoic acid 3-(dimethylamino)propyl 4 (330 mg, 50%), which is a pale yellow oil.

[0577] Synthesis of 4-((4-(bis(2-hydroxydodecyl)amino)butyryl)oxy)-3-methoxybenzoic acid 3-(dimethylamino)propyl ester (VA-3-E12-DMAPr)

[0578]

[0579] 4-((4-(bis(2-((tert-butyldimethylsilyl)oxy)dodecyl)amino)butyryl)oxy)-3-methoxybenzoic acid 3-(dimethylamino)propyl 4 (330 mg, 0.35 mmol) was added dropwise to 10 mL of tetrahydrofuran solution with pyridine (70%, 2.5 mL) containing hydrofluoric acid, and the mixture was stirred overnight at room temperature. Saturated sodium bicarbonate solution was added to pH 7-8, and the mixture was extracted with ethyl acetate. The organic layer was washed with brine and dried over anhydrous sodium sulfate. After filtration and concentration, the crude product was purified by reversed-phase column chromatography (C18: 5-95% MeCN / water / 0.1% TFA) to give 4-((4-(bis(2-hydroxydodecyl)amino)butyryl)oxy)-3-methoxybenzoic acid 3-(dimethylamino)propyl 4 (120 mg, 48%) as a TFA salt. Store this compound in 2-butanol to prevent decomposition.

[0580] All other lipids were prepared in similar yields according to a representative procedure.

[0581] Synthesis scheme of eugenol lipid

[0582]

[0583] Synthesis of 3-(dimethylamino)propyl4-hydroxy-3,5-dimethoxybenzoate (6)

[0584]

[0585] Oxaloyl chloride (12.8 mL, 0.15 mol) was added to a suspension of eugenol 5 (7.5 g, 0.04 mol) in 100 mL of dichloromethane at 0 °C, followed by the addition of dimethylformamide (5 drops), and the resulting mixture was stirred at this temperature for 2 hours. The reaction mixture was evaporated to dryness, and the residue was dissolved in 100 mL of dichloromethane. After cooling to 0 °C, 3-(dimethylamino)prop-1-ol 2 (4.5 mL, 40 mmol) was slowly added, and the reaction mixture was stirred overnight at room temperature. The precipitate was filtered to yield 6-(dimethylamino)propyl 4-hydroxy-3,5-dimethoxybenzoic acid as a white solid (6.2 g, 58%).

[0586] Synthesis of 3-(dimethylamino)propyl 4-((4-(bis(2-((tert-butyldimethylsilyl)oxy)dodecyl)amino)butyryl)oxy)-3,5-dimethoxybenzoic acid (7)

[0587]

[0588] Oxaloyl chloride (0.15 mL, 1.72 mmol) was added to a solution of 4-(bis(2-((tert-butyldimethylsilyl)oxy)dodecyl)amino)butyric acid AIM-3-E12 (0.99 g, 1.41 mmol) in 20 mL of dichloromethane at 0 °C, followed by the addition of dimethylformamide (1 drop), and the mixture was stirred at 0 °C for 2 hours. The reaction mixture was evaporated to dryness, and the residue was dissolved in 20 mL of dichloromethane. After cooling to 0 °C, 6-(dimethylamino)propyl 4-hydroxy-3,5-dimethoxybenzoate (0.2 g, 0.7 mmol) was added, followed by the addition of pyridine (0.35 mL, 4.34 mmol), and the reaction mixture was stirred overnight at room temperature. Ice was added to quench the reaction, and the organic layer was washed with brine and dried over anhydrous sodium sulfate. After filtration and concentration, the crude product was purified by rapid chromatography to obtain 4-((4-(bis(2-((tert-butyldimethylsilyl)oxy)dodecyl)amino)butyryl)oxy)-3,5-dimethoxybenzoic acid 3-(dimethylamino)propyl ester 7 (380 mg, 50%), which is a pale yellow oil.

[0589] Synthesis of 4-((4-(bis(2-hydroxydodecyl)amino)butyryl)oxy)-3,5-dimethoxybenzoic acid 3-(dimethylamino)propyl ester (SA-3-E12-DMAPr)

[0590]

[0591] 380 mg (0.39 mmol) of 4-((4-(bis(2-((tert-butyldimethylsilyl)oxy)dodecyl)amino)butyryl)oxy)-3,5-dimethoxybenzoic acid 3-(dimethylamino)propyl ester was added dropwise to 10 mL of tetrahydrofuran solution at 0 °C with pyridine (70%, 2.5 mL) containing hydrofluoric acid, and the mixture was stirred overnight at room temperature. Saturated sodium bicarbonate solution was added to pH 7-8, and the mixture was extracted with ethyl acetate. The organic layer was washed with brine and dried over anhydrous sodium sulfate. After filtration and concentration, the crude product was purified by reversed-phase column chromatography (C18: 5-95% MeCN / water / 0.1% TFA) to give 4-((4-(bis(2-hydroxydodecyl)amino)butyryl)oxy)-3,5-dimethoxybenzoic acid 3-(dimethylamino)propyl ester (94 mg, 32%) as a TFA salt.

[0592] All other lipids were prepared in similar yields according to a representative procedure.

[0593] Synthesis scheme of sinapic acid lipids

[0594]

[0595] Synthesis of (E)-3-(4-hydroxy-3,5-dimethoxyphenyl)acrylate 3-(dimethylamino)propyl ester (9)

[0596]

[0597] Oxaloyl chloride (7.5 mL, 90 mmol) was added to a suspension of sinapic acid 8 (5 g, 22 mmol) in 100 mL of dichloromethane at 0 °C, followed by the addition of dimethylformamide (5 drops), and the resulting mixture was stirred at this temperature for 2 hours. The reaction mixture was evaporated to dryness, and the residue was dissolved in 100 mL of dichloromethane. After cooling to 0 °C, 3-(dimethylamino)prop-1-ol 2 (2.64 mL, 22 mmol) was slowly added, and the reaction mixture was stirred overnight at room temperature. The precipitate was filtered to yield (E)-3-(4-hydroxy-3,5-dimethoxyphenyl)acrylate 3-(dimethylamino)propyl 9 (2.66 g, 39%) as a pale yellow solid.

[0598] Synthesis of 4-(bis(2-((tert-butyldimethylsilyl)oxy)dodecyl)amino)butyric acid (E)-4-(3-(3-(dimethylamino)propoxy)-3-oxopropyl-1-en-1-yl)-2,6-dimethoxyphenyl ester (10)

[0599]

[0600] Oxaloyl chloride (0.3 mL, 3.09 mmol) was added to a solution of 4-(bis(2-((tert-butyldimethylsilyl)oxy)dodecyl)amino)butyric acid AIM-3-E12 (1.8 g, 2.59 mmol) in 20 mL of dichloromethane at 0 °C, followed by the addition of dimethylformamide (1 drop), and the mixture was stirred at 0 °C for 2 hours. The reaction mixture was evaporated to dryness, and the residue was dissolved in 20 mL of dichloromethane. After cooling to 0 °C, 9 g of (E)-3-(4-hydroxy-3,5-dimethoxyphenyl)acrylate 3-(dimethylamino)propyl ester (0.4 g, 1.29 mmol) was added, followed by the addition of pyridine (0.62 mL, 7.7 mmol), and the reaction mixture was stirred overnight at room temperature. Ice was added to quench the reaction, and the organic layer was washed with brine and dried over anhydrous sodium sulfate. After filtration and concentration, the crude product was purified by rapid chromatography to obtain 10 (540 mg, 42%) of 4-(bis(2-((tert-butyldimethylsilyl)oxy)dodecyl)amino)butyric acid (E)-4-(3-(3-(dimethylamino)propoxy)-3-oxopropyl-1-en-1-yl)-2,6-dimethoxyphenyl ester, which is a pale yellow oil.

[0601] Synthesis of 4-(bis(2-hydroxydodecyl)amino)butyric acid (E)-4-(3-(3-(dimethylamino)propoxy)-3-oxopropyl-1-en-1-yl)-2,6-dimethoxyphenyl ester (SI-3-E12-DMAPr)

[0602]

[0603] At 0 °C, 10 (540 mg, 0.55 mmol) of 4-(bis(2-((tert-butyldimethylsilyl)oxy)dodecyl)amino)butyric acid (E)-4-(3-(3-(dimethylamino)propoxy)-3-oxopropyl-1-en-1-yl)-2,6-dimethoxyphenyl ester 10 was added dropwise to 10 mL of tetrahydrofuran solution, and the mixture was stirred overnight at room temperature. Saturated sodium bicarbonate solution was added to adjust the pH to 7-8, and the mixture was extracted with ethyl acetate. The organic layer was washed with brine and dried over anhydrous sodium sulfate. After filtration and concentration, the crude product was purified by reversed-phase column chromatography (C18: 5-95% MeCN / water / 0.1% TFA) to obtain 4-(bis(2-hydroxydodecyl)amino)butyric acid (E)-4-(3-(3-(dimethylamino)propoxy)-3-oxopropyl-1-en-1-yl)-2,6-dimethoxyphenyl ester (330 mg, 80%) as a TFA salt.

[0604] All other lipids were prepared in similar yields according to a representative procedure.

[0605] Synthesis scheme A of phenolic lipids

[0606]

[0607] Synthesis of intermediate (3a) in synthetic scheme A:

[0608]

[0609] Oxalate chloride (0.98 mL, 4.0 equivalence) was added dropwise to a solution of (1) (2.00 g, 2.86 mmol) in anhydrous CH2Cl2 (10 mL) at 0 °C. The reaction mixture was then slowly heated to room temperature and stirred for 2 hours. Excess solvent and oxalate chloride were removed under reduced pressure, and the remaining residue was redissolved in anhydrous CH2Cl2 (30 mL). (2a) (555 mg, 1.0 equivalence), DMAP (349 mg, 1.0 equivalence), and then triethylamine (3.18 mL, 8.0 equivalence) were added to the stirred acyl chloride solution at 0 °C. The reaction mixture was slowly warmed to room temperature and then stirred at the same temperature for 16 hours. After 16 hours, the reaction mixture was diluted with CH2Cl2 and saturated with NaHCO3. 3(溶液) Solution washing. The separated organic layer was washed with brine, dried over Na2SO4, and concentrated under reduced pressure to obtain the crude product material. The crude product material was first purified with hexane containing 50% EtOAc, and then further purified with CH2Cl2 containing 15% EtOAc to obtain (3a) (623 mg, 25%) as a viscous oil. C60 C60 calculated by MS (ESI+) 50 H 93 NO7Si2, [M+H] + =876.65, observed value =876.6.

[0610] Synthesis of intermediate (3b) in synthetic scheme A:

[0611]

[0612] (2b) followed the procedure of (3a) to provide (3b) as a viscous oil (557 mg, 23%). C calculated by MS (ESI+) 49 H 91 NO6Si2, [M+H] + =846.64, observed value =846.6.

[0613] Synthesis of intermediate (5a) in synthetic scheme A:

[0614]

[0615] Oxaloyl chloride (0.15 mL, 5.0 equivalents) was added to a solution of (3a) (308 mg, 0.351 mmol) in anhydrous CH2Cl2 (3 mL) at room temperature, and the mixture was stirred for 2 hours at the same temperature. Excess solvent and oxaloyl chloride were removed under reduced pressure, and the remaining residue was redissolved in anhydrous CH2Cl2 (3 mL). 3-Dimethylaminopropanol (4) (109 mg, 3 equivalents) was added to the stirred acyl chloride solution at 0 °C, followed by triethylamine (0.10 mL, 2.0 equivalents). The reaction mixture was slowly warmed to room temperature and then stirred at the same temperature for 16 hours. After the reaction was complete as monitored by MS, the reaction mixture was concentrated to dryness under reduced pressure and purified using CH2Cl2 containing 0–10% MeOH to obtain (5a) (204 mg, 60%) as a viscous oil. C5 was calculated by MS (ESI+). 55 H 104 N₂O₇Si₂, [M+H] + =961.74, observed value =961.7.

[0616] Synthesis of intermediate (5b) in synthetic scheme A:

[0617]

[0618] (3b) followed the procedure of (5a) to provide (5b) as a viscous oil (248 mg, 90%). C calculated by MS (ESI+) 54 H 102 N₂O₆Si₂, [M+H] + =931.73, observed value =931.7.

[0619] Synthesis of compound 355(6a) of TBL-0731 in synthetic scheme A:

[0620]

[0621] Pyridine (1.09 mL, 197 equivalents) containing 70% hydrogen fluoride was added dropwise to a stirred solution of (5a) (204 mg, 0.212 mmol) in anhydrous THF (3 mL) at 0 °C, followed by slow heating to room temperature. The reaction mixture was stirred at room temperature for 16 hours. After the reaction was complete as monitored by MS, the reaction mixture was cooled to 0 °C and quenched by the addition of solid NaHCO3 in batches. After minimizing gas formation, the resulting mixture was diluted with EtOAc and quenched with saturated NaHCO3. 3(水溶液)Solution neutralization. The separated organic layer was washed with brine, dried over Na2SO4, and concentrated under reduced pressure to obtain the crude product material. The crude product material was purified using CH2Cl2 containing 0-20% MeOH to obtain TBL-0731(6a) (132 mg, 85%) as a viscous oil. C60 was calculated by MS (ESI+). 43 H 76 N₂O₇, [M+H] + =733.57, Observed value =733.5. 1 H NMR(500MHz, CDCl3)δ7.64(d,J=15.9Hz,1H),7.14–7.08(m,2H),7.05(d,J=8.1, 3.5Hz,1H),6.37(d,J=16.0Hz,1H),4.27(t,J=6.4Hz,2H),3.86(s,3H),3.72–3.6 3(m,2H),2.78–2.70(m,2H),2.67–2.47(m,6H),2.46–2.40(m,2H),2.36(s,6H), 2.02–1.90(m,4H),1.49–1.35(m,4H),1.34–1.15(m,32H),0.87(t,J=6.9Hz,6H).

[0622] Synthesis of compound 467(6b) of TBL-0750 in synthetic scheme A:

[0623]

[0624] TBL-0750(6b) (153 mg, 82%) was supplied as a viscous oil using the procedure (5b) following (6a). CS calculated by MS (ESI+) 42 H 74 N₂O₆, [M+H] + =703.55, observed value =703.6. 1HNMR(400MHz, CDCl3) δ7.66(d,J=16.0Hz,1H),7.53(d,J=8.7,1.3Hz,2H),7.12( d,J=8.6,1.5Hz,2H),6.38(d,J=16.0,0.9Hz,1H),4.26(t,J=6.4Hz,2H),3.70–3. 63(m,2H),2.73–2.64(m,2H),2.64–2.46(m,6H),2.45–2.40(m,2H),2.34(s,6H), 1.99–1.88(m,4H),1.43–1.34(m,4H),1.31–1.21(m,32H),0.87(t,J=6.8Hz,6H).

[0625] Synthesis scheme B of phenolic lipids

[0626]

[0627] Synthesis of intermediate (3) in synthesis scheme B:

[0628]

[0629] Oxaloyl chloride (1.04 mL, 2 equivalents) was added to a solution of acid intermediate (1) (4.58 g) in CH2Cl2 (30 mL, anhydrous) and stirred at room temperature for 2 hours. All volatiles were removed under reduced pressure, and the remaining residue was redissolved in CH2Cl2 (30 mL, anhydrous). Syringic acid (2) (1.32 g, 1.1 equivalents) was then added to this solution, followed by pyridine (2.93 mL, 6 equivalents), and the resulting mixture was stirred overnight at room temperature. After stirring overnight, the solvent was removed under reduced pressure, and the remaining residue was dissolved in a minimal amount of CH2Cl2 and filtered through a short silica stopper using 100% EtOAc as the eluent. The clarified filtrate was concentrated to dryness to provide a crude product material, which was purified by MPLC using a 10-100% EtOAc / hexane gradient via 10CV to provide acid product (3) (3.70 g, 78%). CS calculated by MS (ESI+) 53 H 101 NO8Si2, [M+H] + =936.7, Observed value =936.7.

[0630] Synthesis of intermediate (5a) in synthetic scheme B:

[0631]

[0632] Oxaloyl chloride (0.18 mL, 5 equivalents) was added to a solution of benzoic acid (3) (400 mg) in CH2Cl2 (4 mL, anhydrous) and stirred at room temperature for 2 hours. All volatiles were removed under reduced pressure and the remaining residue was redissolved in CH2Cl2 (3 mL, anhydrous). The resulting solution was cooled in an ice bath and then 2-aminoethanol (4a) (76 mg) was added to a solution of CH2Cl2 (1 mL). The reaction mixture was warmed to room temperature and stirred at the same temperature for 16 hours. After the starting material was completely consumed as monitored by LC-MS, the reaction mixture was concentrated to dryness and the crude product material was purified by MPLC using a 0-20% MeOH / CH2Cl2 gradient via 12CV to provide (5a) (225 mg, 52%) as a viscous oil. C60 was calculated by MS (ESI+). 57 H 110 N₂O₈Si₂, [M+H] + =1007.8, observed value =1007.6.

[0633] Synthesis of intermediate (5b) in synthetic scheme B:

[0634]

[0635] Using 4-aminobutanol (4b) following the synthetic procedure (5a) to provide the product as a viscous oil (5b) (360 mg, 81%). C60 was calculated by MS (ESI+). 59 H 114 N₂O₈Si₂, [M+H] + =1035.8, observed value =1035.6.

[0636] Synthesis of intermediate (5c) in synthetic scheme B:

[0637]

[0638] The synthetic procedure of (5a) was followed using intermediate (3) (500 mg) and 3-morpholinopropanol (4c) to provide product (5c) (350 mg, 62%). C2 was calculated by MS (ESI+). 60 H 114 N₂O₉Si₂, [M+H] + =1063.8, observed value =1063.6.

[0639] Synthesis of intermediate (5d) in synthetic scheme B:

[0640]

[0641] The synthetic procedure (5a) was followed using intermediate (3) (500 mg) and 2-pyridinemethanol (4d) to provide product (5d) (237 mg, 43%). C5 was calculated by MS (ESI+). 60 H 108 N₂O₈Si₂, [M+H] + =1041.8, observed value =1041.6.

[0642] Synthesis of intermediate (5e) in synthetic scheme B:

[0643]

[0644] The synthetic procedure of 5a was followed using intermediate (3) (843 mg) and 4-methylpiperazine ethanol (4e) to provide product (5e) (346 mg, 36%). C2 was calculated by MS (ESI+). 60 H 115 N3O8Si2, [M+H] + =1062.8, observed value =1062.7.

[0645] Synthesis of compound 48(6a) of TBL-0507 in synthetic scheme B:

[0646]

[0647] Triethylamine (0.16 mL, 5 equivalents) was added dropwise to a stirred solution of TBS-protected intermediate (5a) (225 mg) in THF (3 mL, anhydrous) within a plastic polymer scintillation bottle (non-glass), followed by the dropwise addition of triethylamine-3HF (0.36 mL, 10 equivalents). The reaction mixture was stirred overnight at 50 °C. The reaction was carried out by reacting EtOAc with NaHCO3. 3(水溶液) Two drops of the reaction mixture were suspended between the layers to monitor the reaction and analyze the organic layer (TLC or LC-MS). Once the starting material was consumed, excess HF and volatiles were removed by purging with N2 gas in a fume hood, and the remaining material was diluted with EtOAc and neutralized with a saturated aqueous solution of NaHCO3 (checked with pH paper). The separated organic layer was washed with brine, dried with Na2SO4, and concentrated under reduced pressure to provide the crude product material. The crude product material was purified by MPLC at 10 CV using a 0–40% MeOH / CH2Cl2 gradient to provide TBL-0507(6a) (90 mg, 52%). C60 was calculated by MS (ESI+). 45 H 82 N₂O₈, [M+H] + =779.6, observed value =779.5.

[0648] Synthesis of compound 49(6b) of TBL-0508 in synthetic scheme B:

[0649]

[0650] The intermediate (5b) (360 mg) protected by TBS was followed according to procedure (6a) to provide TBL-0508 (6b) (71 mg, 25%). C0 was calculated by MS (ESI+). 47 H 86 N₂O₈, [M+H] + =807.6, observed value =807.5.

[0651] Synthesis of compound 562(6c) of TBL-0517 in synthetic scheme B:

[0652]

[0653] The intermediate (5c) (350 mg) protected by TBS was followed in procedure (6a) and purified using a 0–10% MeOH / CH2Cl2 gradient to provide TBL-0517 (6c) (164 mg, 60%). CL calculated by MS (ESI+) 48 H 86 N₂O₉, [M+H] + =835.6, observed value =835.5.

[0654] Synthesis of compound 563(6d) of TBL-0518 in synthetic scheme B:

[0655]

[0656] The intermediate (5d) (237 mg) protected with TBS was followed in procedure (6a) and purified using a 0–10% MeOH / CH2Cl2 gradient to provide TBL-0518 (6d) (111 mg, 60%). CS calculated by MS (ESI+) 48 H 80 N₂O₈, [M+H] + =813.6, observed value =813.5.

[0657] Synthesis of compound 564(6e) of TBL-0535 in synthetic scheme B:

[0658]

[0659] The intermediate (5e) (346 mg) protected by TBS was purified using the procedure (6a) and purified with a 0–20% MeOH / CH2Cl2 gradient to provide TBL-0535 (6e) (88 mg, 32%). C0 was calculated by MS (ESI+). 48 H 87 N3O8, [M+H] + =834.6, Observed value =834.6.

[0660] Synthesis scheme C for phenolic lipids

[0661]

[0662] Synthesis of intermediate (9a) in synthetic scheme C:

[0663]

[0664] Isopropanol (5 mL) and triethylamine (1.35 mL, 2 equivalents) were added to a flask containing amino acid (7) (500 mg) and dodecyl acrylate (8a) (2.91 g, 2.5 equivalents). The resulting mixture was heated at 90 °C for 3 hours. After the reaction was complete as monitored by MS, the reaction mixture was cooled to room temperature and concentrated under reduced pressure. The remaining material was purified by MPLC using 0–12% MeOH / CH2Cl2 to provide product (9a) (987 mg, 35%). CS calculated by MS (ESI+) 34 H 65 NO. 6, [M+H] + =584.5, Observed value =584.5.

[0665] Synthesis of intermediate (9b) in synthetic scheme C:

[0666]

[0667] The product (9b) (1.80 g, 29%) was prepared using amino acid (7) (1.00 g), tetradecyl acrylate (8b) (6.51 g, 2.5 equivalents), isopropanol (10 mL), and triethylamine (2.70 mL, 2 equivalents) following the procedure in (9a). CS was calculated by MS (ESI+). 38 H 73 NO. 6, [M+H] + =640.5, Observed value =640.5.

[0668] Synthesis of compound 565(11a) of TBL-0484 in synthetic scheme C:

[0669]

[0670] Oxaloyl chloride (1.0 mL, 23 equivalences) was added dropwise to a solution of intermediate (9a) (300 mg) in anhydrous CH2Cl2 (3 mL) at room temperature, and the mixture was stirred for 2 hours at the same temperature. Excess solvent and oxaloyl chloride were removed under reduced pressure, and the remaining residue was redissolved in anhydrous CH2Cl2 (3 mL). Phenolic acid (10) (146 mg, 1.0 equivalences) and pyridine (0.21 mL, 5 equivalences) were added to the stirred acyl chloride solution at room temperature, and the mixture was stirred for 16 hours at the same temperature. After the reaction was monitored by MS for completion, the reaction mixture was concentrated under reduced pressure. The remaining crude product material was purified by MPLC using 0–10% MeOH / CH2Cl2 to provide product TBL-0484 (11a) (90 mg, 21%). CS calculated by MS (ESI+) 48 H 84 N2O 10 [M+H] + =849.6, observed value =849.5.

[0671] Synthesis of compound 566(11b) of TBL-0485 in synthetic scheme C:

[0672]

[0673] TBL-0485 (11b) (80 mg, 19%) was provided using intermediate (9b) (300 mg) following the procedure of (11a). CS was calculated by MS (ESI+). 52 H 92 N2O 10 [M+H] + =905.7, observed value =905.6.

[0674] Example 1: Lipid nanoparticle formulations

[0675] The cationic lipids described herein can be used to prepare lipid nanoparticles according to methods known in the art. For example, suitable methods include those described in International Publication No. WO 2018 / 089801, the entire text of which is incorporated herein by reference.

[0676] An exemplary method for lipid nanoparticle formulations is Method A of WO 2018 / 089801 (see, for example, Example 1 and Example 2 of WO 2018 / 089801). Figure 1Method A (“A”) relates to a conventional method for encapsulating mRNA by mixing it with a lipid mixture without first pre-forming the lipids into lipid nanoparticles. In an exemplary method, an ethanol lipid solution and a buffered aqueous solution of mRNA are prepared separately. A solution of the lipid mixture (cationic lipids, cofactor lipids, zwitterionic lipids, PEG lipids, etc.) is prepared by dissolving the lipids in ethanol. An mRNA solution is prepared by dissolving the mRNA in citrate buffer. The mixture is then heated to 65°C before mixing. The two solutions are then mixed using a pump system. In some cases, a gear pump system is used to mix the two solutions. In some embodiments, a 'T' manifold (or a 'Y' manifold) is used to mix the two solutions. The mixture is then purified by percolation using a TFF method. The resulting formulation is concentrated and stored at 2–8°C until further use.

[0677] A second exemplary method for the formulation of lipid nanoparticles is Method B of WO 2018 / 089801 (see, for example, Example 2 and Figure 2 of WO 2018 / 089801). Method B (“B”) refers to the process of encapsulating messenger RNA (mRNA) by mixing pre-formed lipid nanoparticles with mRNA. A range of different conditions can be used in Method B, such as varying temperatures (i.e., heating or not heating the mixture), buffers, and concentrations. In the exemplary method, lipids dissolved in ethanol and citrate buffer are mixed using a pump system. The instantaneous mixing of the two streams results in the formation of empty lipid nanoparticles, a self-assembly process. The resulting formulation mixture consists of empty lipid nanoparticles in citrate buffer containing alcohol. The formulation is then subjected to a TFF purification process, in which buffer exchange occurs. The resulting suspension of pre-formed empty lipid nanoparticles is then mixed with mRNA using a pump system. For some cationic lipids, heating the solution after mixing results in a higher percentage of mRNA-containing lipid nanoparticles and a higher total mRNA yield.

[0678] The lipid nanoparticle formulations listed in Table 5 were prepared using either method A or B. Each formulation comprised mRNA (FFL mRNA) encoding firefly luciferase protein and lipids (cationic lipids: DMG-PEG2000; cholesterol: DOPE) at mol% ratios listed in Table 5.

[0679] Table 5. Exemplary lipid nanoparticle formulations for intratracheal administration

[0680]

[0681]

[0682] FFL mRNA delivered via intratracheal administration

[0683] By means of A single intratracheal aerosol administration was performed to administer the lipid nanoparticle formulation containing FFL mRNA (50 μL / animal) as shown in Table 5 to anesthetized male CD1 mice (6–8 weeks old). Approximately 24 hours after administration, animals were given 150 mg / kg (60 mg / ml) of luciferin via intraperitoneal injection at 2.5 ml / kg. All animals were imaged using an IVIS imaging system 5–15 minutes later to measure luciferase production in the lungs. Figure 1 This study demonstrates the efficient in vivo delivery of FFL mRNA by lipid nanoparticles comprising cationic lipids as described herein, based on positive luciferase activity.

[0684] Numbered Examples

[0685] 1. A cationic lipid having a structure according to formula (I):

[0686]

[0687] Where L1 is a bond, (C1-C6)alkyl or (C2-C6)alkenyl;

[0688] Where X is O or S;

[0689] Where R 1 R 2 R 3 R 4 and R 5 Each is independently selected from H, OH, optionally substituted (C1-C6)alkyl, optionally substituted (C2-C6)alkenyl, optionally substituted (C2-C6)ynyl, optionally substituted (C1-C6)alkoxy and -OC(O)R';

[0690] Where R 1 R 2 R 3 R 4 or R 5 At least one of them is -OC(O)R';

[0691] Where R' is

[0692]

[0693] Where R 6 for

[0694]

[0695] Where m and p are each independently 0, 1, 2, 3, 4 or 5;

[0696] Where R7 Selected from H, optionally substituted (C1-C6)alkyl, optionally substituted (C2-C6)alkenyl, optionally substituted (C2-C6)ynyl, optionally substituted (C1-C6)acyl, -(CH2). k R A Or -(CH2) k CH(OR 11 )R A ;

[0697] Where R 8 Selected from H, optionally substituted (C1-C6)alkyl, optionally substituted (C2-C6)alkenyl, optionally substituted (C2-C6)ynyl, optionally substituted (C1-C6)acyl, -(CH2). n R B Or -(CH2) n CH(OR 12 )R B ;

[0698] Where R 9 Selected from H, optionally substituted (C1-C6)alkyl, optionally substituted (C2-C6)alkenyl, optionally substituted (C2-C6)ynyl, optionally substituted (C1-C6)acyl, -(CH2). q R C Or -(CH2) q CH(OR 13 )R C ;

[0699] Where R 10 Selected from H, optionally substituted (C1-C6)alkyl, optionally substituted (C2-C6)alkenyl, optionally substituted (C2-C6)ynyl, optionally substituted (C1-C6)acyl, -(CH2). r R D Or -(CH2) r CH(OR 14 )R D ;

[0700] Where k, n, q and r are each 1, 2, 3, 4 or 5 independently;

[0701] Or one of them (i)R 7 and R 8 Or (ii)R 9 and R 10 Together they form optionally substituted 5- or 6-membered heterocyclic alkyl or heteroaryl groups, wherein the heterocyclic alkyl or heteroaryl group comprises 1 to 3 heteroatoms selected from N, O and S;

[0702] Where R 11 R 12 R13 and R 14 Each is independently selected from H, methyl, ethyl, or propyl;

[0703] Where R A R B R C and R D Each is independently selected from the arbitrarily substituted (C6-C) 20 )alkyl, optionally substituted (C6-C 20 alkenyl, optionally substituted (C6-C) 20 ) ynyl group, optionally substituted (C6-C 20 Acyl, optionally substituted -OC(O)alkyl, optionally substituted -OC(O)alkenyl, optionally substituted (C1-C6)monoalkylamino, optionally substituted (C1-C6)dialkylamino, optionally substituted (C1-C6)alkoxy, -OH, -NH2;

[0704] Where R 7 R 8 R 9 R 10 At least one of them includes R A R B R C or R D Part, wherein the R A R B R C or R D Independently selected from optional substitutions (C6-C) 20 )alkyl, optionally substituted (C6-C 20 alkenyl, optionally substituted (C6-C) 20 ) ynyl group, optionally substituted (C6-C 20 Acyl group, optionally substituted -OC(O)(C6-C 20 )alkyl or optionally substituted -OC(O)(C6-C 20 alkenyl;

[0705] Or its pharmaceutically acceptable salt.

[0706] 2. The cationic lipid or a pharmaceutically acceptable salt thereof according to Example 1 with numbering, wherein any alkyl, alkenyl, alkynyl, acyl, alkoxy, monoalkylamino, dialkylamino, heterocyclic alkyl, or heteroaryl group is optionally substituted by one or more substituents selected from the group consisting of: (C1-C6)alkyl, (C2-C6)alkenyl, (C2-C6)alkynyl, (C1-C6)acyl, (C1-C6)alkoxy, halogen, -COR, -CO2H, -CO2R, -CN, -OH, -OR, -OCOR, -OCO2R, -NH2, -NHR, -N(R)2, -SR, or -SO2R, or the two twin hydrogens on the carbon atom are substituted by the group =NH, wherein each instance of R is independently C1-C 10 Aliphatic alkyl groups.

[0707] 3. The cationic lipid or a pharmaceutically acceptable salt thereof according to any one of the foregoing numbered examples, wherein:

[0708] i)R A and R B Same; and / or

[0709] ii)R C and R D same.

[0710] 4. The cationic lipid or a pharmaceutically acceptable salt thereof according to Example 1 or 2 with numbering, wherein:

[0711] i)R A and R B Different; and / or

[0712] ii)R C and R D different.

[0713] 5. A cationic lipid or a pharmaceutically acceptable salt thereof according to any one of the numbered Examples 1 to 3, wherein R A R B R C and R D same.

[0714] 6. A cationic lipid or a pharmaceutically acceptable salt thereof according to any one of the numbered Examples 1 to 4, wherein R A R B R C and R D One or more of them may differ.

[0715] 7. The cationic lipid or a pharmaceutically acceptable salt thereof according to any of the preceding numbered examples, wherein X is O.

[0716] 8. A cationic lipid or a pharmaceutically acceptable salt thereof according to any one of the numbered Examples 1 to 6, wherein X is S.

[0717] 9. A cationic lipid or a pharmaceutically acceptable salt thereof according to any one of the foregoing numbered examples, wherein R 1 R 2 R 3 R 4 and R 5 Only one of them is -OC(O)R'.

[0718] 10. The cationic lipid or a pharmaceutically acceptable salt thereof according to Example 9 (numbered), wherein R 1 R 2 R 3 R 4 or R 5 Neither of them are OH.

[0719] 11. A cationic lipid or a pharmaceutically acceptable salt thereof according to any one of the numbered Examples 1 to 8, wherein R 1 R 2 R 3 R 4 and R 5 The two in the equation are -OC(O)R'.

[0720] 12. The cationic lipid or a pharmaceutically acceptable salt thereof according to Example 11 (numbered), wherein R 1 R 2 R 3 R 4 or R 5 Neither of them are OH.

[0721] 13. A cationic lipid or a pharmaceutically acceptable salt thereof according to any one of the numbered Examples 1 to 8, wherein R 1 R 2 R 3 R 4 and R 5 The three are -OC(O)R'.

[0722] 14. A cationic lipid or a pharmaceutically acceptable salt thereof according to any one of the foregoing numbered examples, wherein R 1 and / or R 5 It is -OC(O)R'.

[0723] 15. A cationic lipid or a pharmaceutically acceptable salt thereof according to any one of the foregoing numbered examples, wherein R 2 and / or R 4It is -OC(O)R'.

[0724] 16. A cationic lipid or a pharmaceutically acceptable salt thereof according to any one of the foregoing numbered examples, wherein R 3 It is -OC(O)R'.

[0725] 17. The cationic lipid or a pharmaceutically acceptable salt thereof according to any one of the preceding numbered examples, wherein:

[0726] i) p, q, and r are the same; or

[0727] ii) One or more of p, q, and r are different; or

[0728] iii) q and r are the same, but p is different.

[0729] 18. The cationic lipid or a pharmaceutically acceptable salt thereof according to any one of the foregoing numbered examples, wherein:

[0730] i) k, m, and n are the same; or

[0731] ii) One or more of k, m, and n are different; or

[0732] iii) k and n are the same, but m is different.

[0733] 19. A cationic lipid or a pharmaceutically acceptable salt thereof according to any of the preceding numbered examples, wherein m is 1, 2 or 3.

[0734] 20. A cationic lipid or a pharmaceutically acceptable salt thereof according to any of the preceding numbered examples, wherein p is 1, 2 or 3.

[0735] 21. The cationic lipid or a pharmaceutically acceptable salt thereof according to any one of the foregoing numbered examples, wherein R' is:

[0736]

[0737] 22. The cationic lipid or a pharmaceutically acceptable salt thereof according to Example 21 (numbered), wherein:

[0738] i) k, m, and n = 1; or

[0739] ii) k, m, and n = 1, and R 11 and R 12 =H; or

[0740] iii) k and n = 1, and m = 2; or

[0741] iv) k and n = 1, m = 2, and R11 and R 12 =H; or

[0742] v)k and n = 1, and m = 3; or

[0743] vi) k and n = 1, m = 3, and R 11 and R 12 =H.

[0744] 23. A cationic lipid or a pharmaceutically acceptable salt thereof according to any one of the foregoing numbered examples, wherein R 6 for:

[0745]

[0746] 24. The cationic lipid or a pharmaceutically acceptable salt thereof according to Example 23 (numbered), wherein:

[0747] i) p, q, and r = 1; or

[0748] ii) p, q and r = 1, and R 13 and R 14 For H; or

[0749] iii) q and r = 1, and p = 2; or

[0750] iv) q and r = 1, p = 2, and R 13 and R 14 For H.

[0751] 25. A cationic lipid or a pharmaceutically acceptable salt thereof according to any one of the numbered Examples 1 to 22, wherein R 6 Choose from the following groups:

[0752]

[0753] 26. The cationic lipid or a pharmaceutically acceptable salt thereof according to Example 25 with numbering, wherein R 6 for:

[0754]

[0755] 27. The cationic lipid according to any one of the foregoing numbered examples, having a structure according to formula (II):

[0756]

[0757] Or its pharmaceutically acceptable salt.

[0758] 28. The cationic lipid according to Example 27 (numbered), having a structure according to formula (IIA):

[0759]

[0760] Or its pharmaceutically acceptable salt.

[0761] 29. The cationic lipid according to Example 28 (numbered), having a structure according to one of formulas (IIB), (IIC), (IID), or (IIE):

[0762]

[0763] Or its pharmaceutically acceptable salt.

[0764] 30. The cationic lipid according to Example 27 (numbered), having a structure according to formula (IIF):

[0765]

[0766] Or its pharmaceutically acceptable salt.

[0767] 31. The cationic lipid according to Example 27 (numbered), having a structure according to formula (IIG):

[0768]

[0769] Or its pharmaceutically acceptable salt.

[0770] 32. The cationic lipid according to Example 27 (numbered), having a structure according to formula (IIH):

[0771]

[0772] One of Y and Z is OH and the other is -OC(O)R', or both Y and Z are independently -OC(O)R', or their pharmaceutically acceptable salts.

[0773] 33. The cationic lipid according to any one of the numbered Examples 1 to 26, having a structure according to formula (III):

[0774]

[0775] Or its pharmaceutically acceptable salt.

[0776] 34. The cationic lipid according to Example 33 (numbered), having a structure according to formula (IIIA):

[0777]

[0778] Or its pharmaceutically acceptable salt.

[0779] 35. The cationic lipid according to Example 33 or Example 34, which has a structure according to formula (IIIB):

[0780]

[0781] Or its pharmaceutically acceptable salt.

[0782] 36. The cationic lipid according to Example 35 (numbered), having a structure according to formula (IIIC):

[0783]

[0784] Or its pharmaceutically acceptable salt.

[0785] 37. The cationic lipid according to Example 33 (numbered), having a structure according to formula (IIID):

[0786]

[0787] Or its pharmaceutically acceptable salt.

[0788] 38. The cationic lipid according to Example 37 (numbered), having a structure selected from formulas (IIIE), (IIIF), (IIIG), (IIIH), (IIII), (IIIJ), or (IIIK):

[0789]

[0790]

[0791]

[0792] Or its pharmaceutically acceptable salt.

[0793] 39. The cationic lipid according to Example 33 (numbered), having a structure according to formula (IIIL):

[0794]

[0795] Or its pharmaceutically acceptable salt.

[0796] 40. The cationic lipid according to Example 33 (numbered), having a structure according to formula (IV):

[0797]

[0798] M is selected from H, OH, OMe or Me, or a pharmaceutically acceptable salt thereof.

[0799] 41. The cationic lipid according to Example 33 (numbered), having a structure according to formula (VI), (VII), (VIII), (IX) or (X):

[0800]

[0801]

[0802] One of Y and Z is OH and the other is -OC(O)R', or both Y and Z are independently -OC(O)R', or their pharmaceutically acceptable salts.

[0803] 42. The cationic lipid or a pharmaceutically acceptable salt thereof according to Example 41 with numbering, wherein one of Y and Z is OH and the other is -OC(O)R'.

[0804] 43. The cationic lipid or a pharmaceutically acceptable salt thereof according to Example 42 with numbering, wherein Y is OH and Z is -OC(O)R'.

[0805] 44. The cationic lipid or a pharmaceutically acceptable salt thereof according to Example 42 with numbering, wherein Y is -OC(O)R' and Z is OH.

[0806] 45. The cationic lipid or a pharmaceutically acceptable salt thereof according to Example 41 with numbering, wherein both Y and Z are -OC(O)R'.

[0807] 46. ​​A compound selected from the compounds listed in Tables 1 to 8, or a pharmaceutically acceptable salt thereof.

[0808] 47. A composition comprising any of the preceding numbered examples, a cationic lipid, one or more non-cationic lipids, one or more cholesterol-based lipids, and one or more PEG-modified lipids.

[0809] 48. The composition according to Example 47 with numbering, wherein the composition is lipid nanoparticles, optionally liposomes.

[0810] 49. The composition according to Example 48 (numbered), wherein the one or more cationic lipids constitute about 30 mol% to 60 mol% of the lipid nanoparticles.

[0811] 50. The composition according to any one of the numbered Examples 48 or 49, wherein the one or more non-cationic lipids constitute 10 mol%-50 mol% of the lipid nanoparticles.

[0812] 51. The composition according to any one of the numbered Examples 48 to 50, wherein the one or more PEG-modified lipids constitute 1 mol% to 10 mol% of the lipid nanoparticles.

[0813] 52. The composition according to any one of the numbered Examples 48 to 51, wherein the cholesterol-based lipid constitutes 10 mol% to 50 mol% of the lipid nanoparticles.

[0814] 53. The composition according to any one of the numbered Examples 48 to 52, wherein the lipid nanoparticles encapsulate nucleic acids, optionally mRNA encoding peptides or proteins.

[0815] 54. The composition according to any one of the numbered Examples 48 to 52, wherein the lipid nanoparticles encapsulate mRNA encoding peptides or proteins.

[0816] 55. The composition according to Example 54 (numbered), wherein the lipid nanoparticles encapsulate at least 70% of the mRNA.

[0817] 56. The composition according to Example 54 (numbered), wherein the lipid nanoparticles encapsulate at least 75% of the mRNA.

[0818] 57. The composition according to Example 54 (numbered), wherein the lipid nanoparticles encapsulate at least 80% of the mRNA.

[0819] 58. The composition according to Example 54 (numbered), wherein the lipid nanoparticles encapsulate at least 85% of the mRNA.

[0820] 59. The composition according to Example 54 (numbered), wherein the lipid nanoparticles encapsulate at least 90% of the mRNA.

[0821] 60. The composition according to Example 54 (numbered), wherein the lipid nanoparticles encapsulate at least 95% of the mRNA.

[0822] 61. The composition according to any one of the numbered Examples 54 to 60, used for treatment.

[0823] 62. A method of treating or preventing a disease suitable for treatment or prevention by means of a peptide or protein encoded by mRNA, according to any one of the numbered Examples 54 to 60, wherein the disease is optionally: (a) a protein deficiency, wherein the protein deficiency optionally affects the liver, lungs, brain or muscles; (b) an autoimmune disease; (c) an infectious disease; or (d) cancer.

[0824] 63. The composition for use according to Example 61 or 62 with numbering, wherein the composition is administered intravenously, intrathecally or intramuscularly, or delivered via the lungs, optionally by nebulization.

[0825] 64. A method for treating or preventing a disease, wherein the method comprises administering to a subject in need a composition of any one of the numbered Examples 54 to 60, and wherein the disease is suitable for treatment or prevention by means of a peptide or protein encoded by mRNA, optionally wherein the disease is: (a) a protein deficiency, optionally wherein the protein deficiency affects the liver, lungs, brain or muscles; (b) an autoimmune disease; (c) an infectious disease; or (d) cancer.

[0826] 65. The method according to Example 64 with number, wherein the composition is administered intravenously, intrathecally, or intramuscularly, or delivered via the lungs, optionally by nebulization.

Claims

1. A cationic lipid having a structure according to formula (I): Where L1 is a bond, (C1-C6)alkyl or (C2-C6)alkenyl; Where X is O or S; Where R 1 R 2 R 3 R 4 and R 5 Each is independently selected from H, OH, optionally substituted (C1-C6)alkyl, optionally substituted (C2-C6)alkenyl, optionally substituted (C2-C6)ynyl, optionally substituted (C1-C6)alkoxy and -OC(O)R'; Where R 1 R 2 R 3 R 4 or R 5 At least one of them is -OC(O)R'; Where R' is Where R 6 for Where m and p are each independently 0, 1, 2, 3, 4 or 5; Where R 7 Selected from H, optionally substituted (C1-C6)alkyl, optionally substituted (C2-C6)alkenyl, optionally substituted (C2-C6)ynyl, optionally substituted (C1-C6)acyl, -(CH2). k R A Or -(CH2) k CH(OR 11 )R A ; Where R 8 Selected from H, optionally substituted (C1-C6)alkyl, optionally substituted (C2-C6)alkenyl, optionally substituted (C2-C6)ynyl, optionally substituted (C1-C6)acyl, -(CH2). n R B Or -(CH2) n CH(OR 12 )R B ; Where R 9 Selected from H, optionally substituted (C1-C6)alkyl, optionally substituted (C2-C6)alkenyl, optionally substituted (C2-C6)ynyl, optionally substituted (C1-C6)acyl, -(CH2). q R C Or -(CH2) q CH(OR 13 )R C ; Where R 10 Selected from H, optionally substituted (C1-C6)alkyl, optionally substituted (C2-C6)alkenyl, optionally substituted (C2-C6)ynyl, optionally substituted (C1-C6)acyl, -(CH2). r R D Or -(CH2) r CH(OR 14 )R D ; Where k, n, q and r are each 1, 2, 3, 4 or 5 independently; Or one of them (i)R 7 and R 8 Or (ii)R 9 and R 10 Together they form optionally substituted 5- or 6-membered heterocyclic alkyl or heteroaryl groups, wherein the heterocyclic alkyl or heteroaryl group comprises 1 to 3 heteroatoms selected from N, O and S; Where R 11 R 12 R 13 and R 14 Each is independently selected from H, methyl, ethyl, or propyl; Where R A R B R C and R D Each is independently selected from the arbitrarily substituted (C6-C) 20 )alkyl, optionally substituted (C6-C 20 alkenyl, optionally substituted (C6-C) 20 ) ynyl group, optionally substituted (C6-C 20 Acyl, optionally substituted -OC(O)alkyl, optionally substituted -OC(O)alkenyl, optionally substituted (C1-C6)monoalkylamino, optionally substituted (C1-C6)dialkylamino, optionally substituted (C1-C6)alkoxy, -OH, -NH2; Where R 7 R 8 R 9 R 10 At least one of them includes R A R B R C or R D Part, wherein the R A R B R C or R D Independently selected from optional substitutions (C6-C) 20 )alkyl, optionally substituted (C6-C 20 alkenyl, optionally substituted (C6-C) 20 ) ynyl group, optionally substituted (C6-C 20 Acyl group, optionally substituted -OC(O)(C6-C 20 )alkyl or optionally substituted -OC(O)(C6-C 20 alkenyl; Or its pharmaceutically acceptable salt.

2. The cationic lipid of claim 1 or a pharmaceutically acceptable salt thereof, wherein X is O.

3. The cationic lipid or a pharmaceutically acceptable salt thereof according to claim 1 or claim 2, wherein m is 1, 2 or 3.

4. The cationic lipid or a pharmaceutically acceptable salt thereof according to any one of the preceding claims, wherein p is 1, 2 or 3.

5. The cationic lipid or a pharmaceutically acceptable salt thereof according to any one of the preceding claims, wherein R ' for:

6. The cationic lipid of claim 5 or a pharmaceutically acceptable salt thereof, wherein: vii)k, m and n = 1; or viii) k, m and n = 1, and R 11 and R 12 =H; or ix)k and n=1, and m=2; or x)k and n=1, m=2, and R 11 and R 12 =H; or xi)k and n=1, and m=3; or xii)k and n=1, m=3, and R 11 and R 12 =H.

7. The cationic lipid or a pharmaceutically acceptable salt thereof according to any one of claims 1 to 4, wherein R ' for And R 7 and R 8 Each for itself - CO2R aa The substituted (C1-C6) alkyl group is optionally substituted, wherein R aa For C1-C 50 Alkyl, preferably wherein R 7 and R 8 Each for itself - CO2R aa Substituted (C1-C6) alkyl groups, wherein R aa C1-C 20 Alkyl, more preferably wherein R 7 and R 8 Each is:

8. The cationic lipid or a pharmaceutically acceptable salt thereof according to any one of the preceding claims, wherein R 6 for:

9. The cationic lipid according to any one of the preceding claims, having a structure according to the following: (a) Formula (II): Or its pharmaceutically acceptable salt. (b) Equation (IIA): Or its pharmaceutically acceptable salt. (c) Equation (IIF): Or its pharmaceutically acceptable salt. (d) Formula (IIG): Or its pharmaceutically acceptable salt. (e) Equation (IIH): Where one of Y and Z is OH and the other is -OC(O)R', or where both Y and Z are independently -OC(O)R', or their pharmaceutically acceptable salts. (f) Equation (III): Or a pharmaceutically acceptable salt thereof, (g) formula (IIIA): Or its pharmaceutically acceptable salt, (h) formula (IIIB): Or a pharmaceutically acceptable salt thereof, (i) formula (IIID): Or its pharmaceutically acceptable salt. (j) Equation (IIIL): Or its pharmaceutically acceptable salt, or (k) Equation (IV): M is selected from H, OH, OMe or Me, or a pharmaceutically acceptable salt thereof.

10. The cationic lipid according to any one of claims 1 to 8, having a structure according to formula (VI), (VII), (VIII), (IX) or (X): One of Y and Z is OH and the other is -OC(O)R', or both Y and Z are independently -OC(O)R', or their pharmaceutically acceptable salts.

Images