Novel homocysteine-based lipids and their use for delivering nucleic acids

Abstract

Lipids are disclosed that can be used to deliver and express mRNA and encoded proteins, e.g., as components of liposome delivery vehicles, and accordingly can be used to treat various diseases, disorders, and conditions, such as those associated with deficiency of one or more proteins.

Description

Background Technology

[0001] Nucleic acid delivery has been extensively explored as a potential therapeutic option for certain disease states. In particular, messenger RNA (mRNA) therapy has become an increasingly important option for treating a variety of diseases, including those associated with deficiencies in one or more proteins.

[0002] Lipid nanoparticles (LNPs) are spherical vesicles made of ionizable lipids that are positively charged at low pH (to enable RNA complexation) and neutral at physiological pH (reducing potential toxicity compared to positively charged lipids such as liposomes). LPNs are considered attractive carriers for therapeutics and remain a subject of ongoing development.

[0003] There is a need to continuously improve the transfection of nucleic acids (such as mRNA). Summary of the Invention

[0004] In particular, this invention provides homocysteine-based lipids that can be used to deliver mRNA. The delivery of mRNA provided by the lipids described herein can result in targeted delivery, reduced administration frequency, improved patient tolerability, and provide more effective and less toxic mRNA vaccines or therapies for the prevention and / or treatment of a variety of diseases, including but not limited to influenza and respiratory syncytial virus (RSV) vaccines.

[0005] In one objective, the present invention provides a compound having formula (I):

[0006] (I)

[0007] in

[0008] m is an integer selected from 1 to 6; preferably 2 to 4;

[0009] n is an integer selected from 1 to 6; preferably 2 to 4;

[0010] p is an integer selected from 1 to 6; preferably 2 to 4;

[0011] Same or different R 1 and R 2 Independently selected from the group consisting of: straight-chain or branched (C1-C30) alkyl and straight-chain or branched (C2-C30) alkenyl.

[0012] Each of the alkyl and alkenyl groups is optionally separated by one or more groups selected from the following: –C=O-, -C=OO-, and –O-, and / or

[0013] Each of the alkyl and alkenyl groups is optionally substituted by one or more substituents selected from the following: –OR, -CN-, -(C1-C6)alkyl-OH, -CF3, -NO2, -COOR, -SR, halogen atom and -NRR';

[0014] R 3 The group consisting of (C1-C6) alkyl groups selected from H, optionally substituted with one or more of the following substituents: –OR, -CN-, -(C1-C6)alkyl-OH, -CF3, -NO2, -COOR, -SR, halogen atom and -NRR';

[0015] Same or different R 4 and R 5 Independently selected from the group consisting of: straight-chain or branched (C1-C6) alkyl and straight-chain or branched (C2-C6) alkenyl, each alkyl or alkenyl optionally substituted by one or more substituents selected from the group consisting of: –OR, -CN-, -(C1-C6)alkyl-OH, -CF3, -NO2, -COOR, -SR, halogen atom and -NRR';

[0016] Or R 4 and R 5 Together with the N atoms to which they are attached, they form the following:

[0017] Five- to six-membered cycloalkyl or heterocyclic compounds containing one to four heteroatoms selected from O, N, and S, or

[0018] Five- to six-membered aryl or heteroaryl groups containing one to four heteroatoms selected from O, N, and S.

[0019] The cycloalkyl, heterocyclic, aryl, or heteroaryl groups may optionally be substituted by one or more substituents selected from the following: -(C1-C6)alkyl, –OR, -CN-, -(C1-C6)alkyl-OH, -CF3, -NO2, -COOR, -SR, halogen atom, and -NRR';

[0020] Same or different R 6 and R 7 Independently selected from the group consisting of: straight-chain or branched (C1-C30) alkyl and straight-chain or branched (C2-C30) alkenyl.

[0021] Each of the alkyl and alkenyl groups is optionally separated by one or more groups selected from the following: –C=O-, -C=OO-, and –O-, and / or

[0022] Each of the alkyl and alkenyl groups is optionally substituted by one or more substituents selected from the following: –OR, -CN-, -C1-C6 alkyl-OH, -CF3, -NO2, -COOR, -SR, halogen atom and -NRR';

[0023] Same or different R 8 and R 9 Independently select the group composed of H and -OR;

[0024] --- indicates an optional single bond, provided that, when present, compound (I) behaves as if formed by X. - The positive charge that counteracts the ion balance + The form represents the quaternary ammonium form, and when it is absent, compound (I) is in a neutral form and X is absent;

[0025] X - These are anions that can be present at will;

[0026] R 10 Choose from the group consisting of H and straight-chain or branched (C1-C30) alkyl groups;

[0027] The same or different R and R' are independently selected from H and (C1-C6) alkyl groups;

[0028] Or its pharmaceutically acceptable salts or esters.

[0029] In another objective, the invention is further characterized by a method for preparing a compound having formula (I), the method comprising the steps of: taking a compound having formula (II):

[0030] (II)

[0031] Reaction with compounds having formula (III):

[0032] (III)

[0033] Where m, n, p, R 1 R 2 R 3 R 4 R 5 R 6 and R 7 Defined in this article;

[0034] or

[0035] The method includes the following one-pot step: preparing a compound having formula (III):

[0036] (III)

[0037] Compounds having formula (IV)

[0038] (IV)

[0039] Compounds having formula (V):

[0040] (V)

[0041] Reaction with compounds having formula (VI):

[0042] (VI)

[0043] Where m, n, p, R 1 R 2 R 3 R 4 R 5 R 6 and R 7 Defined in this article.

[0044] In a further objective, the present invention also relates to a lipid nanoparticle (LNP), such as a liposome, said LNP comprising a compound having formula (I) as defined herein, and optionally encapsulating nucleic acids.

[0045] For yet another purpose, the present invention relates to a pharmaceutical composition comprising an LNP as defined herein, and one or more pharmaceutically acceptable excipients.

[0046] In still other purposes, the present invention also relates to LNPs as defined herein for delivering nucleic acids into the cells of a patient, particularly as vaccines, such as vaccines against influenza or respiratory syncytial virus (RSV) vaccines. Detailed Implementation

[0047] definition

[0048] To facilitate understanding of the invention, certain terms are defined below. Further definitions of the following and other terms are set forth throughout the specification. Publications and other references cited herein to describe the background of the invention and to provide further details about its implementation are hereby incorporated by reference.

[0049] 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 universal 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 l-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 (such as amides), and / or substitutes. Amino acids (including carboxyl-terminal 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 association 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, 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. It is obvious from the context in which the term is used whether it refers to free amino acids or peptide residues.

[0050] 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, a non-human animal is a mammal (e.g., rodents, mice, rats, rabbits, monkeys, dogs, cats, sheep, cattle, primates, and / or pigs). In some embodiments, an animal includes, but is not limited to, mammals, birds, reptiles, amphibians, fish, insects, and / or worms. In some embodiments, an animal may be a transgenic animal, a genetically engineered animal, and / or a clone.

[0051] Approximately or about: As used herein, the term “approximately” or “about” when applied to one or more target values ​​refers to a value similar to the stated 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 of the stated reference value in either direction, unless otherwise stated or otherwise apparent from the context (except where such a number would exceed 100% of the possible value).

[0052] 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 administered to that organism is considered biologically active.

[0053] Delivery: As used herein, the term “delivery” encompasses both local delivery and systemic delivery. For example, mRNA delivery encompasses the delivery of mRNA to a target tissue and the expression and retention of the encoded protein within that target tissue (also known as “local distribution” or “local delivery”), as well as the delivery of mRNA to a target tissue and the expression and secretion of the encoded protein into the patient’s circulatory system (e.g., serum) and its systemic distribution and uptake by other tissues (also known as “systemic distribution” or “systemic delivery”).

[0054] Expression: As used herein, “expression” of a nucleic acid sequence refers to the translation of mRNA into a polypeptide, the assembly of multiple polypeptides into a complete protein (e.g., an enzyme), and / or the post-translational modification of a polypeptide or a fully assembled protein (e.g., an enzyme). In this application, the terms “expression” and “production”, as well as grammatical equivalents, are used interchangeably.

[0055] Functionality: As used herein, a “functional” biomolecule is a biomolecule that exhibits the properties and / or activities that characterize it.

[0056] 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 measured at the beginning of a time period.

[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, which is, for example, 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 subject” is a subject with the same form of disease as the subject being treated and who is approximately the same age as the subject being treated.

[0058] In vitro: As used herein, the term “in vitro” refers to events that occur in an artificial environment (e.g., in test tubes or reaction vessels, in cell cultures, etc.) rather than within a multicellular organism.

[0059] In vivo: As used herein, the term "in vivo" refers to events that occur 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 that occur 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 has been separated from at least some of the components associated with it during initial production (whether in nature and / or in an experimental setting) and / or (2) by hand production, preparation, and / or manufacture. The separated substance and / or entity may be separated from other components that are approximately 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or more than 99% of their initial association. In some embodiments, the separated agent is approximately 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or more than 99% pure. As used herein, a substance is considered "pure" if it contains substantially no other components. As used herein, the calculation of the percentage purity of isolated substances and / or entities 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, liposomes as used herein can be formed by mixing one or more lipids or by mixing one or more lipids with one or more 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 both modified 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 a recombinant expression system and optionally purified, chemically synthesized, etc. Where appropriate, for example, 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 indicated, mRNA sequences are presented in a 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-propynyl-cytidine, C-5-propynyl-uridine, 2-aminoadenosine, C5-bromouridine, C5-fluorouridine, C5-iodouridine, C5-propynyl-uridine, C5-propynyl-cytidine, C5-methylcytidine). , 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., thiophosphate and 5'-N-phosphoramide linked).

[0063] Nucleic acid: As used herein, the term “nucleic acid” in its broadest sense refers to any compound and / or substance incorporated into or potentially incorporated into a polynucleotide chain. In some embodiments, a nucleic acid is a compound and / or substance incorporated into a polynucleotide chain via or potentially via a phosphodiester bond. In some embodiments, “nucleic acid” refers to a single nucleic acid residue (e.g., a 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), multimeric coding nucleic acid (MCNA), polymeric coding nucleic acid (PCNA), guide RNA (gRNA), and CRISPR RNA (crRNA). In some embodiments, "nucleic acid" encompasses deoxyribonucleic acid (DNA), including but not limited to any one or more of single-stranded DNA (ssDNA), double-stranded DNA (dsDNA), and complementary DNA (cDNA). In some embodiments, "nucleic acid" encompasses both RNA and DNA. In embodiments, DNA may be in the form of antisense DNA, plasmid DNA, 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 in the form of 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 Cahalosome-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 non-coding RNA (lncRNA), microRNA (miRNA), piwi-interacting RNA (piRNA), small interfering RNA (siRNA), transcribed 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 term "patient" or "subject" means any biological organism to which the provided composition may be administered, 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 both prenatal and postnatal forms.

[0065] Pharmaceutically acceptable: As used herein, the term “pharmaceutically acceptable” means, to the extent of reasonable medical judgment, a substance suitable for use in contact with human and animal tissues without excessive toxicity, irritation, allergic reactions, or other problems or complications commensurate with a reasonable benefit / risk ratio.

[0066] Pharmaceutically acceptable salts: Pharmaceutically acceptable salts are well known in the art. For example, SM Berge et al. described pharmaceutically acceptable salts in detail in J. 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 salts formed by reacting an amino group with an inorganic acid (such as hydrochloric acid, hydrobromic acid, phosphoric acid, sulfuric acid, and perchloric acid) or with an organic acid (such as acetic acid, oxalic acid, maleic acid, tartaric acid, citric acid, succinic acid, or malonic acid), or by using other methods used in the art (such as ion exchange). Other pharmaceutically acceptable salts include adipate, alginate, ascorbate, aspartate, benzenesulfonate, benzoate, hydrogen sulfate, borate, butyrate, camphorate, camphorsulfonate, citrate, cyclopentanepropionate, digluconate, dodecyl sulfate, ethanesulfonate, formate, fumarate, glucono-heptate, glyceryl phosphate, gluconate, hemisulfate, heptaate, hydroiodate, 2-hydroxy-ethanesulfonate, lacturonate, lactate, laurate, lauryl sulfate, malate, maleate, malonate, methanesulfonate, 2-naphthalenesulfonate, nicotinate, nitrate, oleate, oxalate, palmitate, dihydroxynaphthalate, pectate, persulfate, 3-phenylpropionate, phosphate, picrate, pentanoate, 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, magnesium, etc. Where appropriate, other pharmaceutically acceptable salts include non-toxic ammonium, quaternary ammonium, and amine cations formed using counterions (such as halide, hydroxide, carboxyl, sulfate, phosphate, nitrate, sulfonate, and arylsulfonate). Other pharmaceutically acceptable salts include those formed by quaternizing amines with suitable electrophilic agents (e.g., alkyl halides) to form quaternized alkylamino salts.

[0067] Whole-body distribution or delivery: As used herein, the terms “whole-body distribution,” “whole-body delivery,” or their grammatical equivalents refer to a mechanism or method of delivery or distribution that affects the whole body or the whole organism. Typically, whole-body distribution or delivery is accomplished via the body’s circulatory system (e.g., blood flow). Compare this to 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 subject is a human being. A subject can be a patient, referring to a person who goes to a healthcare provider for diagnosis or treatment of a disease. The term "subject" is used interchangeably herein with "individual" or "patient." A subject may have or be susceptible to a disease or disorder, but may or may not exhibit symptoms of the disease or disorder.

[0069] Essentially: As used herein, the term “essentially” refers to a qualitative condition that exhibits all or nearly all of the intended characteristics or properties, or to a degree or extent. Those skilled in the art of biology will understand that biological and chemical phenomena rarely (if at all) complete and / or proceed to completion or achieve or avoid an absolute result. Therefore, the term “essentially” is used herein to capture the potential lack of completeness inherent 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 pathology, symptoms, or features associated with the disease.

[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 one or more symptoms of a disease, disorder, and / or condition when administered to a subject who has or is susceptible to such disease, disorder, and / or condition. Those skilled in the art will understand 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, relieve, reduce, suppress, prevent, or delay the onset, severity, and / or 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 for the purpose of reducing the risk of developing disease-related pathology.

[0073] Aliphatic: As used in this article, the term aliphatic refers to C 1- C 40 Hydrocarbons, 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 groups), C2-C 20Alkenyl (e.g., straight-chain or branched C4-C) 20 Dieneyl, 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 compounds can include C3-C 20 Cyclic aliphatic (e.g., C3-C) 20 cycloalkyl, C4-C 20 Cycloalkenyl, or C8-C 20 Cycloalkyne group). 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 substituent groups as described herein. For example, the aliphatic group may be substituted with one or more of halogen, -COR', -CO2H, -CO2R', -CN, -OH, -OR', -OCOR', -OCO2R', -NH2, -NHR', -N(R')2, -SR', or -SO2R' (e.g., 1, 2, 3, 4, 5, or 6 independently selected substituents), wherein each example 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 (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.

[0074] Alkyl: As used herein, the term "alkyl" refers to acyclic straight-chain and branched hydrocarbon groups, such as "C1-C". 20"Alkyl" refers to an alkyl group having 1 to 20 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. Other alkyl groups will be apparent to those skilled in the art in light of the benefits of this disclosure. Alkyl groups can be unsubstituted or substituted with one or more substituent groups as described herein. For example, an alkyl group can be substituted with one or more of halogen, -COR', -CO2H, -CO2R', -CN, -OH, -OR', -OCOR', -OCO2R', -NH2, -NHR', -N(R')2, -SR', or -SO2R' (e.g., 1, 2, 3, 4, 5, or 6 independently selected substituents), wherein each example 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 (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., 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 a “hydroxyalkyl” group, wherein the prefix indicates a –OH group and “alkyl” is as described herein.

[0075] Alkylene: As used herein, the term "alkylene" refers to a saturated divalent straight-chain or branched hydrocarbon group, such as methylene, ethylene, isopropylene, etc. Similarly, the term "alkenylene" as used herein 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 on the chain; while the term "alkynylene" as used 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 on 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, the 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 example 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 (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.

[0076] Alkenyl: As used herein, "alkenyl" means any straight or branched hydrocarbon chain having one or more unsaturated carbon-carbon double bonds that can appear at any stable point on the chain, such as "C2-C". 20"Alkenyl" refers to an alkenyl group having 2-20 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 concatenated. The alkenyl group may be unsubstituted. Or it may be substituted by one or more substituent groups as described herein. For example, the alkenyl group may be substituted by one or more of halogen, -COR', -CO2H, -CO2R', -CN, -OH, -OR', -OCOR', -OCO2R', -NH2, -NHR', -N(R')2, -SR', or -SO2R' (e.g., 1, 2, 3, 4, 5, or 6 independently selected substituents), wherein each example 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 (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., 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 a “hydroxyalkenyl” group, wherein the prefix indicates a –OH group and “alkenyl” is as described herein.

[0077] Alkynyl: As used herein, "alkynyl" means any straight-chain or branched hydrocarbon chain having one or more carbon-carbon triple bonds, which can be present at any stable point on the chain, such as "C2-C". 20"Alynyl" refers to an alkynyl group having 2 to 20 carbons. 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 examples, the alkynyl group contains a carbon-carbon triple bond. The alkynyl group may be unsubstituted or substituted with one or more substituent groups as described herein. For example, the alkynyl group may be substituted with one or more of halogen, -COR', -CO2H, -CO2R', -CN, -OH, -OR', -OCOR', -OCO2R', -NH2, -NHR', -N(R')2, -SR', or -SO2R' (e.g., 1, 2, 3, 4, 5, or 6 independently selected substituents), wherein each example 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 (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., 1, 2, 3, 4, 5, or 6 substituent groups as described herein).

[0078] Aryl: The terms “aryl” and “aromatic”, used alone or as part of a larger part, such as “arylalkyl,” “arylalkoxy,” or “aryloxyalkyl,” refer to a C-shaped compound containing one to three aromatic rings with optional substitutions. 6–14 Aromatic hydrocarbon moiety. For example, the aryl group is C. 6–10 Aryl groups (i.e., phenyl and naphthyl). Aryl groups include, but are not limited to, optionally substituted phenyl, naphthyl, or anthracene groups. As used herein, the terms “aryl” and “aromatic” also include groups in which the aryl ring is fused with one or more cyclic aliphatic rings to form optionally substituted cyclic structures such as tetrahydronaphthyl, indenyl, or indenyl rings. The term “aryl” may be used interchangeably with the terms “aryl group,” “aryl ring,” and “aromatic ring.”

[0079] Cycloalkyl: As used herein, the term "cycloalkyl" refers to a non-aromatic saturated cyclic group, such as "C3-C". 10"Cycloalkyl". In the examples, the cycloalkyl group is monocyclic. In the examples, the cycloalkyl group is polycyclic (e.g., bicyclic or tricyclic). In polycyclic cycloalkyl groups, the single ring can be fused, bridged, or spirocyclic. Examples of cycloalkyl groups include cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, norbornel, bicyclo[3.2.1]octyl, octahydro-cyclopentadienyl, and spiro[4.5]decyl, etc. The term "cycloalkyl" may be used interchangeably with the term "carbocyclic". The cycloalkyl group can be Unsubstituted or substituted with one or more substituent groups as described herein. For example, the cycloalkyl group may be substituted with one or more of halogen, -COR', -CO2H, -CO2R', -CN, -OH, -OR', -OCOR', -OCO2R', -NH2, -NHR', -N(R')2, -SR', or -SO2R' (e.g., 1, 2, 3, 4, 5, or 6 independently selected substituents), wherein each example 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 (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 cycloalkyl is unsubstituted. In the examples, the cycloalkyl is substituted (e.g., 1, 2, 3, 4, 5, or 6 substituent groups as described herein).

[0080] Halogen: As used in this article, the term “halogen” means fluorine, chlorine, bromine or iodine.

[0081] Heteroalkenyl. The term "heteroalkenyl" refers to a branched or unbranched alkenyl group having 2 to 14 carbon atoms in addition to 1, 2, 3, or 4 heteroatoms independently selected from the group consisting of N, O, S, and P. Heteroalkenyl groups may optionally include monocyclic, bicyclic, or tricyclic rings, wherein each ring preferably has three to six members. Heteroalkenyl groups may be substituted or unsubstituted.

[0082] Zeoyynyl. The term "zeoyynyl" refers to a branched or unbranched ynyl group having 2 to 14 carbon atoms in addition to 1, 2, 3, or 4 heteroatoms independently selected from the group consisting of N, O, S, and P. Zeoyynyl groups may optionally include monocyclic, bicyclic, or tricyclic rings, wherein each ring preferably has three to six members. Zeoyynyl groups may be substituted or unsubstituted.

[0083] Heteroalkyl. The term "heteroalkyl" means a branched or unbranched alkyl group having 1 to 14 carbon atoms in addition to 1, 2, 3, or 4 heteroatoms independently selected from the group consisting of N, O, S, and P. Heteroalkyl groups include, but are not limited to, tertiary amines, secondary amines, ethers, thioethers, amides, thioamides, carbamates, thiocarbamates, hydrazones, imines, phosphate diesters, phosphoramides, sulfonamides, and disulfides. Heteroalkyl groups may optionally include monocyclic, bicyclic, or tricyclic rings, wherein each ring is preferably composed of three to six members. Heteroalkyl groups may be substituted or unsubstituted. Examples of heteroalkyl groups include, but are not limited to, polyethers such as methoxymethyl and ethoxyethyl.

[0084] Heteroaryl: The terms “heteroaryl” and “heteroaryl-”, used alone or as part of a larger group, such as “heteroarylalkyl” or “heteroarylalkoxy,” refer to a group that: has 5 to 14 ring atoms (preferably 5, 6, 9, or 10 ring atoms); has 6, 10, or 14 π electrons shared in a cyclic array; and has one to five heteroatoms in addition to a carbon atom. Heteroaryl groups can be monocyclic, bicyclic, tricyclic, or polycyclic, for example, monocyclic, bicyclic, or tricyclic (e.g., monocyclic or bicyclic). The term “heteroatom” refers to nitrogen, oxygen, or sulfur, and includes any oxidized form of nitrogen or sulfur, as well as any quaternized form of basic nitrogen. For example, the nitrogen atom of a heteroaryl group can be a basic nitrogen atom and can optionally be oxidized to the corresponding N-oxide. When a heteroaryl group is substituted with a hydroxyl group, it also includes its corresponding tautomer. As used herein, the terms “heteroaryl” and “heteroary-” also include groups in which a heteroaryl ring is fused with one or more aryl, cycloaliphatic or heterocyclic aliphatic rings. Non-limiting examples of heteroaryl groups include thiophene, furanyl, pyrrole, imidazolyl, pyrazolyl, triazolyl, tetrazolyl, oxazolyl, isoxazolyl, oxadiazolyl, thiazolyl, isothiazolyl, thiazolyl, pyridinyl, pyrazinyl, indoleazinyl, purine, naphridinyl, pteridinyl, indole, isoindole, benzothiophene, benzofuranyl, dibenzofuranyl, inzolyl, benzimidazolyl, benzothiazolyl, quinolinyl, isoquinolinyl, terolinyl, phthalazinyl, quinazolinyl, quinoxolinyl, 4H-quinazinyl, carbazolyl, acridineyl, phenazinyl, phenothiazinyl, phenothiazinyl, tetrahydroquinolinyl, tetrahydroisoquinolinyl and pyrido[2,3-b]-1,4-oxazin-3(4H)-one. The term "heteroaryl" may be used interchangeably with the terms "heteroaryl ring," "heteroaryl group," or "heteroaromatic," any of which includes an optionally substituted ring. The term "heteroarylalkyl" refers to an alkyl group substituted with a heteroaryl group, wherein the alkyl and heteroaryl portions are optionally substituted independently.

[0085] Heterocyclic group. As used herein, the terms “heterocycle,” “heterocyclic group,” “heterocyclic radical,” and “heterocyclic ring” are used interchangeably and refer to a stable 3- to 8-membered monocyclic or 7- to 10-membered bicyclic heterocyclic portion that is saturated or partially unsaturated and has one or more (e.g., one to four) heteroatoms as defined above, in addition to a carbon atom. When used to refer to the ring atom of a heterocycle, the term “nitrogen” includes substituted nitrogen. As an example, in a saturated or partially unsaturated ring having 0–3 heteroatoms selected from oxygen, sulfur, or nitrogen, nitrogen can be N (e.g., in 3,4-dihydro-2H-pyrrole), NH (e.g., in pyrrolealkyl), or NR. + (e.g., in N-substituted pyrrolidinyl groups).

[0086] Heterocycles can have their side groups attached at any heteroatom or carbon atom that results in a stable structure, and any one of the ring atoms can optionally be substituted. Examples of such saturated or partially unsaturated heterocyclic radicals include, but are not limited to, tetrahydrofuranyl, tetrahydrothiophenyl, piperidinyl, decahydroquinolinyl, oxazolidinyl, piperazine, dioxane, dioxopentyl, diazazyl, oxonitrilezyl, thioazinotrilezyl, morpholinyl, and thiomorpholinyl. The heterocyclic group can be monocyclic, bicyclic, tricyclic, or polycyclic, preferably monocyclic, bicyclic, or tricyclic, more preferably monocyclic or bicyclic. The term "heterocyclic alkyl" refers to an alkyl group substituted with a heterocyclic group, wherein the alkyl and heterocyclic portions are optionally substituted independently. Additionally, heterocycles also include groups in which the heterocycle is fused with one or more aryl rings.

[0087] The lipids of the present invention

[0088] While liposome-based mediators containing lipid components have shown promising results in terms of encapsulation, stability, and site localization, lipid-based delivery systems still require significant improvement. For example, major drawbacks of liposome delivery systems involve the construction of liposomes that are sufficiently stable in cell culture or in vivo to reach the desired target cells and / or intracellular compartments, and the ability of such liposome delivery systems to efficiently release their encapsulating material into these target cells.

[0089] In particular, there remains a need for improved lipids that exhibit improved pharmacokinetic properties and are capable of delivering macromolecules such as nucleic acids to a variety of cell types and tissues with enhanced efficiency. Importantly, there is also a particular need for novel lipids characterized by reduced toxicity and the ability to efficiently deliver encapsulated nucleic acids and polynucleotides to target cells, tissues, and organs.

[0090] This document describes novel lipids, compositions comprising such lipids, and related uses. In the examples, the compounds described herein can be used as LPNs to facilitate delivery to one or more target cells and subsequent transfection of one or more target cells.

[0091] The lipids disclosed herein contain basic ionizable functional groups (e.g., amines or nitrogen-containing heteroaryl groups as described herein) existing in neutral or charged forms. For example, a basic ionizable functional group may refer to a nitrogen-containing functional group (e.g., NH2, guanidine, amidine, monoalkylamine or dialkylamine, 5- to 6-membered heterocyclic alkyl, or 5- to 6-membered nitrogen-containing heteroaryl group), which can be converted into a charged group by protonation with an acid or deprotonation with a base.

[0092] In the embodiments, compounds having formula (I) as described herein may provide one or more desired features or properties. That is, in some embodiments, the lipids described herein may be characterized by having one or more properties that provide an advantage over other lipids of similar classification. For example, the lipids disclosed herein may allow control and customization of the properties of LPNs in which they are components. In particular, the lipids disclosed herein may be characterized by enhanced transfection efficiency and their ability to elicit specific biological outcomes. Such outcomes may include, for example, enhanced cellular uptake, endosome / lysosome disruption, and / or promotion of the release of intracellular encapsulating materials (e.g., polynucleotides).

[0093] The lipids of the present invention have formula (I):

[0094] (I)

[0095] in

[0096] m is an integer selected from 1 to 6; preferably 2 to 4;

[0097] n is an integer selected from 1 to 6; preferably 2 to 4;

[0098] p is an integer selected from 1 to 6; preferably 2 to 4;

[0099] Same or different R 1 and R 2 Independently selected from the group consisting of: straight-chain or branched (C1-C30) alkyl and straight-chain or branched (C2-C30) alkenyl.

[0100] Each of the alkyl and alkenyl groups is optionally separated by one or more groups selected from the following: –C=O-, -C=OO-, and –O-, and / or

[0101] Each of the alkyl and alkenyl groups is optionally substituted by one or more substituents selected from the following: –OR, -CN-, -(C1-C6)alkyl-OH, -CF3, -NO2, -COOR, -SR, halogen atom and -NRR';

[0102] R 3The group consisting of (C1-C6) alkyl groups selected from H, optionally substituted with one or more of the following substituents: –OR, -CN-, -(C1-C6)alkyl-OH, -CF3, -NO2, -COOR, -SR, halogen atom and -NRR';

[0103] Same or different R 4 and R 5 Independently selected from the group consisting of: straight-chain or branched (C1-C6) alkyl and straight-chain or branched (C2-C6) alkenyl, each alkyl or alkenyl optionally substituted by one or more substituents selected from the group consisting of: –OR, -CN-, -(C1-C6)alkyl-OH, -CF3, -NO2, -COOR, -SR, halogen atom and -NRR';

[0104] Or R 4 and R 5 Together with the N atoms to which they are attached, they form the following:

[0105] Five- to six-membered cycloalkyl or heterocyclic compounds containing one to four heteroatoms selected from O, N, and S, or

[0106] Five- to six-membered aryl or heteroaryl groups containing one to four heteroatoms selected from O, N, and S.

[0107] The cycloalkyl, heterocyclic, aryl, or heteroaryl groups may optionally be substituted by one or more substituents selected from the following: -(C1-C6)alkyl, –OR, -CN-, -(C1-C6)alkyl-OH, -CF3, -NO2, -COOR, -SR, halogen atom, and -NRR';

[0108] Same or different R 6 and R 7 Independently selected from the group consisting of: straight-chain or branched (C1-C30) alkyl and straight-chain or branched (C2-C30) alkenyl.

[0109] Each of the alkyl and alkenyl groups is optionally separated by one or more groups selected from the following: –C=O-, -C=OO-, and –O-, and / or

[0110] Each of the alkyl and alkenyl groups is optionally substituted by one or more substituents selected from the following: –OR, -CN-, -C1-C6 alkyl-OH, -CF3, -NO2, -COOR, -SR, halogen atom and -NRR';

[0111] Same or different R 8 and R 9 Independently select the group composed of H and -OR;

[0112] --- indicates an optional single bond, provided that, when present, compound (I) behaves as if formed by X. - The positive charge that counteracts the ion balance + The form represents the quaternary ammonium form, and when it is absent, compound (I) is in a neutral form and X is absent;

[0113] X - These are anions that can be present at will;

[0114] R 10 Choose from the group consisting of H and straight-chain or branched (C1-C30) alkyl groups;

[0115] The same or different R and R' are independently selected from H and (C1-C6) alkyl groups;

[0116] Or its pharmaceutically acceptable salts or esters.

[0117] The expressions “lipids having formula (I) as described herein” and “compounds having formula (I)” are used interchangeably and still refer to lipids / compounds having formulas (IA)-(II) as disclosed above.

[0118] In the embodiments, in equation (I), the same or different R 1 and R 2 Independently selected from the group consisting of: straight-chain or branched (C1-C30) alkyl and straight-chain or branched (C2-C30) alkenyl.

[0119] Each of the alkyl and alkenyl groups is optionally separated by one or more groups selected from –C=O- and –O-, and / or

[0120] Each of the alkyl and alkenyl groups is optionally substituted by one or more substituents selected from –OH.

[0121] In the embodiments, the same or different R 6 and R 7 Independently selected from the group consisting of: straight-chain or branched (C1-C30) alkyl and straight-chain or branched (C2-C30) alkenyl.

[0122] Each of the alkyl and alkenyl groups is optionally separated by one or more groups selected from –C=O- and –O-.

[0123] In the embodiment, R 3 Select from the group consisting of H and (C1-C6) alkyl groups;

[0124] Same or different R 4 and R 5The group consisting of straight-chain or branched (C1-C6) alkyl groups is independently selected from the group consisting of one or more substituents selected from the group consisting of -OH.

[0125] Or R 4 and R 5 Together with the N atoms to which they are attached, they form the following:

[0126] Five- to six-membered cycloalkyl or heterocyclic compounds containing one to two heteroatoms selected from O, N, and S.

[0127] Five- to six-membered aryl or heteroaryl groups containing one to two heteroatoms selected from O, N, and S.

[0128] The cycloalkyl, heterocyclic, aryl, or heteroaryl groups may optionally be substituted by one or more substituents selected from the following: OH, -(C1-C6)alkyl, and C1-C6alkyl-OH.

[0129] In the embodiment, X - Derived from halogen atoms, such as Cl - ,Br - F - Or I - Alternatively, it can be selected from trifluoroacetate, formate, p-toluenesulfonate, acetate, succinate, and chlorate.

[0130] In the embodiment, R 1 R 2 R 6 and R 7 Each of the following groups is independently selected:

[0131]

[0132]

[0133]

[0134]

[0135] In the embodiment, R 4 and R 5 Independently selected from one of the following groups:

[0136]

[0137] Or they can form any of the following rings together with the N atoms to which they are attached:

[0138]

[0139] The premise is that any of the above rings may optionally be substituted by one or more substituents selected from the following: - –OR, -CN-, -(C1-C6)alkyl-OH, -CF3, -NO2, -COOR, -SR, halogen atom and –NRR', where R and R' are as defined herein.

[0140] In the embodiments, the compound having formula (I) may be selected from those having the following formulas (IA), (I-A'), (I-A''), (IB), (IC), (ID), (IE), (IF), (IG), (IH), (II), or (IJ):

[0141] (IA)

[0142] (I-A')

[0143] (I-A'')

[0144] (IB)

[0145]

[0146] (IC)

[0147]

[0148] (ID)

[0149]

[0150] (IE)

[0151] (IF)

[0152] (IG)

[0153] (IH)

[0154] (II)

[0155]

[0156] (IJ)

[0157] Where m, n, p, R 1 R 2 R 3 R 4 R 5 R 6R 7 R 8 R 9 、- - -、R、R' and optional X、R 10 As defined in this article;

[0158] Furthermore, the same or different q and q' independently represent integers equal to 7, 8, 9, 10, or 11;

[0159] Or its pharmaceutically acceptable salts or esters.

[0160] In the embodiments, the compound having formula (I) may be selected from the group consisting of:

[0161] 18-((2-(diethylamino)ethyl)(methyl)carbamoyl)-7,26-dihydroxy-9-(2-hydroxy-8-(((Z)-non-2-en-1-yl)oxy)-8-oxooctyl)-24-(2-hydroxydodecyl)-20-oxo-14,15-dithia-9,19,24-triazahexadecanoic acid(Z)-non-2-en-1-yl ester

[0162] 18-((2-(diethylamino)ethyl)(methyl)carbamoyl)-7,26-dihydroxy-9-(2-hydroxy-8-(((Z)-non-2-en-1-yl)oxy)-8-oxooctyl)-24-(2-hydroxydodecyl)-20-oxo-14,15-dithia-9,19,24-triazahexadecanoic acid(Z)-non-2-en-1-yl ester

[0163] 18-((2-(diethylamino)ethyl)carbamoyl)-7,26-dihydroxy-9-(2-hydroxy-8-(((Z)-non-2-en-1-yl)oxy)-8-oxooctyl)-24-(2-hydroxydodecyl)-20-oxo-14,15-dithia-9,19,24-triazahexadecanoic acid(Z)-non-2-en-1-yl ester

[0164] N-(1-((3-(1H-imidazol-1-yl)propyl)amino)-4-((4-(bis(2-hydroxytetradecyl)amino)butyl)dithioalkyl)-1-oxobutyl-2-yl)-5-(bis(2-hydroxydecyl)amino)pentanamide)

[0165] 18-((3-(diethylamino)propyl)carbamoyl)-7,26-dihydroxy-9-(2-hydroxy-8-(((Z)-non-2-en-1-yl)oxy)-8-oxooctyl)-24-(2-hydroxydodecyl)-20-oxo-14,15-dithia-9,19,24-triazahexadecanoic acid(Z)-non-2-en-1-yl ester

[0166] 18-((2-(dimethylamino)ethyl)carbamoyl)-7,26-dihydroxy-9-(2-hydroxy-8-(((Z)-non-2-en-1-yl)oxy)-8-oxooctyl)-24-(2-hydroxydodecyl)-20-oxo-14,15-dithia-9,19,24-triazahexadecanoic acid(Z)-non-2-en-1-yl ester

[0167] 18-((2-(dimethylamino)ethyl)(methyl)carbamoyl)-7,26-dihydroxy-9-(2-hydroxy-8-(((Z)-non-2-en-1-yl)oxy)-8-oxooctyl)-24-(2-hydroxydodecyl)-20-oxo-14,15-dithia-9,19,24-triazahexadecanoic acid(Z)-non-2-en-1-yl ester

[0168] 18-((3-(1H-imidazol-1-yl)propyl)carbamoyl)-7,26-dihydroxy-9-(2-hydroxy-8-(((Z)-non-2-en-1-yl)oxy)-8-oxooctyl)-24-(2-hydroxydodecyl)-20-oxo-14,15-dithia-9,19,24-triazahexahexadecanoic acid (Z)-non-2-en-1-yl ester

[0169] 7,26-Dihydroxy-9-(2-hydroxy-8-(((Z)-non-2-en-1-yl)oxy)-8-oxooctyl)-24-(2-hydroxydodecyl)-20-oxo-18-((2-(piperidin-1-yl)ethyl)carbamoyl)-14,15-dithia-9,19,24-triazahexahexadecanoic acid(Z)-non-2-en-1-yl ester

[0170] 7,26-Dihydroxy-9-(2-hydroxy-8-(((Z)-non-2-en-1-yl)oxy)-8-oxooctyl)-24-(2-hydroxydodecyl)-18-((4-morpholinobutyl)carbamoyl)-20-oxo-14,15-dithia-9,19,24-triazahexahexadecanoic acid (Z)-non-2-en-1-yl ester

[0171] 7,25-Dihydroxy-9-(2-hydroxy-8-(((Z)-non-2-en-1-yl)oxy)-8-oxooctyl)-23-(2-hydroxydodecyl)-19-oxo-17-((4-(pyrrolidone-1-yl)butyl)carbamoyl)-14-thia-9,18,23-triazapentadecanoic acid (Z)-non-2-en-1-yl ester

[0172] 5-(bis(2-hydroxydecyl)amino)-N-(19-hydroxy-17-(2-hydroxytetradecyl)-2-methyl-7-oxo-11,12-dithia-2,6,17-triazatrionecan-8-yl)pentanamide

[0173] 5-(bis(2-hydroxydecyl)amino)-N-(3-ethyl-19-hydroxy-17-(2-hydroxytetradecyl)-7-oxo-11,12-dithia-3,6,17-triazatrionecan-8-yl)pentanamide

[0174] 5-(bis(2-hydroxydecyl)amino)-N-(3-ethyl-20-hydroxy-18-(2-hydroxytetradecyl)-8-oxo-12,13-dithia-3,7,18-triazatridodecane-9-yl)pentanamide

[0175] 5-(bis(2-hydroxydecyl)amino)-N-(18-hydroxy-16-(2-hydroxytetradecyl)-2-methyl-6-oxo-10,11-dithia-2,5,16-triazatriacontane-7-yl)pentanamide

[0176] 5-(bis(2-hydroxydecyl)amino)-N-(18-hydroxy-16-(2-hydroxytetradecyl)-2,5-dimethyl-6-oxo-10,11-dithia-2,5,16-triazatriacontane-7-yl)pentanamide

[0177] 5-(bis(2-hydroxydecyl)amino)-N-(3-ethyl-19-hydroxy-17-(2-hydroxytetradecyl)-6-methyl-7-oxo-11,12-dithia-3,6,17-triazatrionecan-8-yl)pentanamide

[0178] 5-(bis(2-hydroxydecyl)amino)-N-(1,19-dihydroxy-17-(2-hydroxytetradecyl)-3-methyl-7-oxo-11,12-dithia-3,6,17-triazatrionecan-8-yl)pentanamide

[0179] 5-(bis(2-hydroxydecyl)amino)-N-(4-((4-(bis(2-hydroxytetradecyl)amino)butyl)dithioalkyl)-1-((4-morpholinobutyl)amino)-1-oxobutyl-2-yl)pentanamide

[0180] 5-(bis(2-hydroxydecyl)amino)-N-(4-((4-(bis(2-hydroxytetradecyl)amino)butyl)dithioalkyl)-1-oxo-1-((4-(pyrrolidone-1-yl)butyl)amino)but-2-yl)pentanamide

[0181] 4-((4-(bis((9Z,12Z)-2-hydroxyoctadec-9,12-dien-1-yl)amino)butyl)dithioalkyl)-2-(4-(bis(2-hydroxydodecyl)amino)butamido)-N-(3-(dimethylamino)propyl)butamido)

[0182] 4-((4-(bis((9Z,12Z)-2-hydroxyoctadec-9,12-dien-1-yl)amino)butyl)dithioalkyl)-2-(4-(bis(2-hydroxydodecyl)amino)butamido)-N-(2-(diethylamino)ethyl)butamido)

[0183] 4-((4-(bis((9Z,12Z)-2-hydroxyoctadec-9,12-dien-1-yl)amino)butyl)dithioalkyl)-2-(4-(bis(2-hydroxydodecyl)amino)butamido)-N-(3-(diethylamino)propyl)butamido)

[0184] 4-((4-(bis((9Z,12Z)-2-hydroxyoctadec-9,12-dien-1-yl)amino)butyl)dithioalkyl)-2-(4-(bis(2-hydroxydodecyl)amino)butamido)-N-(2-(dimethylamino)ethyl)butamido)

[0185] 4-((4-(bis((9Z,12Z)-2-hydroxyoctadec-9,12-dien-1-yl)amino)butyl)dithioalkyl)-2-(4-(bis(2-hydroxydodecyl)amino)butamido)-N-(2-(dimethylamino)ethyl)-N-methylbutamido)

[0186] 4-((4-(bis((9Z,12Z)-2-hydroxyoctadec-9,12-dien-1-yl)amino)butyl)dithioalkyl)-2-(4-(bis(2-hydroxydodecyl)amino)butamido)-N-(2-(diethylamino)ethyl)-N-methylbutamido)

[0187] N-(3-(1H-imidazol-1-yl)propyl)-4-((4-(bis((9Z,12Z)-2-hydroxyoctadec-9,12-dien-1-yl)amino)butyl)dithioalkyl)-2-(4-(bis(2-hydroxydodecyl)amino)butyramide)butyramide

[0188] 4-((4-(bis((9Z,12Z)-2-hydroxyoctadec-9,12-dien-1-yl)amino)butyl)dithioalkyl)-2-(4-(bis(2-hydroxydodecyl)amino)butamido)-N-(4-morpholinobutyl)butamido)

[0189] 4-((4-(bis((9Z,12Z)-2-hydroxyoctadec-9,12-dien-1-yl)amino)butyl)dithioalkyl)-2-(4-(bis(2-hydroxydodecyl)amino)butamido)-N-(4-(pyrrolidone-1-yl)butyl)butamido)

[0190] (Z)-18-((3-(dimethylamino)propyl)carbamoyl)-26-hydroxy-24-(2-hydroxydodecyl)-9-(octadec-9-en-1-yl)-20-oxo-14,15-dithia-9,19,24-triazahexadecanoic acid heptadecanoyl ester

[0191] (Z)-18-((2-(diethylamino)ethyl)carbamoyl)-26-hydroxy-24-(2-hydroxydodecyl)-9-(octadec-9-en-1-yl)-20-oxo-14,15-dithia-9,19,24-triazahexadecanoic acid heptadecanoyl ester

[0192] (Z)-18-((3-(diethylamino)propyl)carbamoyl)-26-hydroxy-24-(2-hydroxydodecyl)-9-(octadec-9-en-1-yl)-20-oxo-14,15-dithia-9,19,24-triazahexadecanoic acid heptadecanoyl ester

[0193] (Z)-18-((2-(dimethylamino)ethyl)carbamoyl)-26-hydroxy-24-(2-hydroxydodecyl)-9-(octadec-9-en-1-yl)-20-oxo-14,15-dithia-9,19,24-triazahexadecanoic acid heptadecanoyl ester

[0194] (Z)-18-((2-(dimethylamino)ethyl)(methyl)carbamoyl)-26-hydroxy-24-(2-hydroxydodecyl)-9-(octadec-9-en-1-yl)-20-oxo-14,15-dithia-9,19,24-triazahexadecanoic acid heptadecanoyl ester

[0195] (Z)-18-((2-(diethylamino)ethyl)(methyl)carbamoyl)-26-hydroxy-24-(2-hydroxydodecyl)-9-(octadec-9-en-1-yl)-20-oxo-14,15-dithia-9,19,24-triazahexadecanoic acid heptadecanoyl ester

[0196] (Z)-18-((3-(1H-imidazol-1-yl)propyl)carbamoyl)-26-hydroxy-24-(2-hydroxydodecyl)-9-(octadec-9-en-1-yl)-20-oxo-14,15-dithia-9,19,24-triazahexadecanoic acid heptadecanyl ester

[0197] (Z)-26-hydroxy-24-(2-hydroxydodecyl)-18-((2-((2-hydroxyethyl)(methyl)amino)ethyl)carbamoyl)-9-(octadec-9-en-1-yl)-20-oxo-14,15-dithia-9,19,24-triazahexadecanoic acid heptadecanyl ester

[0198] (Z)-26-hydroxy-24-(2-hydroxydodecyl)-18-((4-morpholinobutyl)carbamoyl)-9-(octadec-9-en-1-yl)-20-oxo-14,15-dithia-9,19,24-triazahexadecanoic acid heptadecanoyl ester

[0199] (Z)-26-hydroxy-24-(2-hydroxydodecyl)-9-(octadec-9-en-1-yl)-20-oxo-18-((4-(pyrrolidin-1-yl)butyl)carbamoyl)-14,15-dithia-9,19,24-triazahexadecanoic acid heptadecanoyl ester

[0200] 18-((3-(dimethylamino)propyl)carbamoyl)-25-hydroxy-23-(2-hydroxydecyl)-9-(8-(((Z)-non-2-en-1-yl)oxy)-8-oxooctyl)-20-oxo-14,15-dithia-9,19,23-triazatriacontaneanoic acid (Z)-non-2-en-1-yl ester

[0201] 18-((2-(diethylamino)ethyl)carbamoyl)-25-hydroxy-23-(2-hydroxydecyl)-9-(8-(((Z)-non-2-en-1-yl)oxy)-8-oxooctyl)-20-oxo-14,15-dithia-9,19,23-triazatriacontaneanoic acid (Z)-non-2-en-1-yl ester

[0202] 18-((3-(diethylamino)propyl)carbamoyl)-25-hydroxy-23-(2-hydroxydecyl)-9-(8-(((Z)-non-2-en-1-yl)oxy)-8-oxooctyl)-20-oxo-14,15-dithia-9,19,23-triazatriacontaneanoic acid (Z)-non-2-en-1-yl ester

[0203] 18-((2-(dimethylamino)ethyl)carbamoyl)-25-hydroxy-23-(2-hydroxydecyl)-9-(8-(((Z)-non-2-en-1-yl)oxy)-8-oxooctyl)-20-oxo-14,15-dithia-9,19,23-triazatriacontaneanoic acid (Z)-non-2-en-1-yl ester

[0204] 18-((2-(dimethylamino)ethyl)(methyl)carbamoyl)-25-hydroxy-23-(2-hydroxydecyl)-9-(8-(((Z)-non-2-en-1-yl)oxy)-8-oxooctyl)-20-oxo-14,15-dithia-9,19,23-triazatriacontaracanoic acid (Z)-non-2-en-1-yl ester

[0205] 18-((2-(diethylamino)ethyl)(methyl)carbamoyl)-25-hydroxy-23-(2-hydroxydecyl)-9-(8-(((Z)-non-2-en-1-yl)oxy)-8-oxooctyl)-20-oxo-14,15-dithia-9,19,23-triazatriacontaneanoic acid (Z)-non-2-en-1-yl ester

[0206] 18-((3-(1H-imidazol-1-yl)propyl)carbamoyl)-25-hydroxy-23-(2-hydroxydecyl)-9-(8-(((Z)-non-2-en-1-yl)oxy)-8-oxooctyl)-20-oxo-14,15-dithia-9,19,23-triazatriacontaneanoic acid (Z)-non-2-en-1-yl ester

[0207] 25-Hydroxy-23-(2-Hydroxydecyl)-9-(8-(((Z)-Non-2-en-1-yl)oxy)-8-oxooctyl)-20-oxo-18-((2-(piperidin-1-yl)ethyl)glycyl)-14,15-dithia-9,19,23-triazatriacontaracanoic acid (Z)-non-2-en-1-yl ester

[0208] 25-Hydroxy-23-(2-Hydroxydecyl)-18-((4-morpholinobutyl)carbamoyl)-9-(8-(((Z)-non-2-en-1-yl)oxy)-8-oxooctyl)-20-oxo-14,15-dithia-9,19,23-triazatriacontaracanoic acid (Z)-non-2-en-1-yl ester

[0209] 25-Hydroxy-23-(2-Hydroxydecyl)-9-(8-(((Z)-Non-2-en-1-yl)oxy)-8-oxooctyl)-20-oxo-18-((4-(pyrrolidone-1-yl)butyl)carbamoyl)-14,15-dithia-9,19,23-triazatriacontaracanonicoic acid (Z)-non-2-en-1-yl ester

[0210] 18-((3-(dimethylamino)propyl)carbamoyl)-26-hydroxy-24-(2-hydroxy-8-(((Z)-non-2-en-1-yl)oxy)-8-oxooctyl)-9-(8-(((Z)-non-2-en-1-yl)oxy)-8-oxooctyl)-20-oxo-14,15-dithia-9,19,24-triazatridodecanoic acid di((Z)-non-2-en-1-yl) ester

[0211] 18-((2-(diethylamino)ethyl)carbamoyl)-26-hydroxy-24-(2-hydroxy-8-(((Z)-non-2-en-1-yl)oxy)-8-oxooctyl)-9-(8-(((Z)-non-2-en-1-yl)oxy)-8-oxooctyl)-20-oxo-14,15-dithia-9,19,24-triazatridodecanoic acid di((Z)-non-2-en-1-yl) ester

[0212] 18-((3-(diethylamino)propyl)carbamoyl)-26-hydroxy-24-(2-hydroxy-8-(((Z)-non-2-en-1-yl)oxy)-8-oxooctyl)-9-(8-(((Z)-non-2-en-1-yl)oxy)-8-oxooctyl)-20-oxo-14,15-dithia-9,19,24-triazatridodecanoic acid di((Z)-non-2-en-1-yl) ester

[0213] 18-((2-(dimethylamino)ethyl)carbamoyl)-26-hydroxy-24-(2-hydroxy-8-(((Z)-non-2-en-1-yl)oxy)-8-oxooctyl)-9-(8-(((Z)-non-2-en-1-yl)oxy)-8-oxooctyl)-20-oxo-14,15-dithia-9,19,24-triazatridodecanoic acid di((Z)-non-2-en-1-yl) ester

[0214] 18-((2-(dimethylamino)ethyl)(methyl)carbamoyl)-26-hydroxy-24-(2-hydroxy-8-(((Z)-non-2-en-1-yl)oxy)-8-oxooctyl)-9-(8-(((Z)-non-2-en-1-yl)oxy)-8-oxooctyl)-20-oxo-14,15-dithia-9,19,24-triazatridodecanoic acid di((Z)-non-2-en-1-yl) ester

[0215] 18-((2-(diethylamino)ethyl)(methyl)carbamoyl)-26-hydroxy-24-(2-hydroxy-8-(((Z)-non-2-en-1-yl)oxy)-8-oxooctyl)-9-(8-(((Z)-non-2-en-1-yl)oxy)-8-oxooctyl)-20-oxo-14,15-dithia-9,19,24-triazatridodecanoic acid di((Z)-non-2-en-1-yl) ester

[0216] 18-((3-(1H-imidazol-1-yl)propyl)carbamoyl)-26-hydroxy-24-(2-hydroxy-8-(((Z)-non-2-en-1-yl)oxy)-8-oxooctyl)-9-(8-((((Z)-non-2-en-1-yl)oxy)-8-oxooctyl)-20-oxo-14,15-dithia-9,19,24-triazatridodecanoic acid di((Z)-non-2-en-1-yl) ester

[0217] 26-Hydroxy-24-(2-hydroxy-8-(((Z)-non-2-en-1-yl)oxy)-8-oxooctyl)-9-(8-(((Z)-non-2-en-1-yl)oxy)-8-oxooctyl)-20-oxo-18-((2-(piperidin-1-yl)ethyl)glycyl)-14,15-dithia-9,19,24-triazatridodecanoic acid di((Z)-non-2-en-1-yl) ester

[0218] 26-Hydroxy-24-(2-hydroxy-8-(((Z)-non-2-en-1-yl)oxy)-8-oxooctyl)-18-((4-morpholinobutyl)carbamoyl)-9-(8-(((Z)-non-2-en-1-yl)oxy)-8-oxooctyl)-20-oxo-14,15-dithia-9,19,24-triazatridodecanoic acid di((Z)-non-2-en-1-yl) ester

[0219] 26-Hydroxy-24-(2-hydroxy-8-(((Z)-non-2-en-1-yl)oxy)-8-oxooctyl)-9-(8-(((Z)-non-2-en-1-yl)oxy)-8-oxooctyl)-20-oxo-18-((4-(pyrrolidone-1-yl)butyl)carbamoyl)-14,15-dithia-9,19,24-triazatridodecanoic acid di((Z)-non-2-en-1-yl) ester

[0220] 18-((3-(dimethylamino)propyl)carbamoyl)-7,26-dihydroxy-9,24-bis(2-hydroxy-8-(((Z)-non-2-en-1-yl)oxy)-8-oxooctyl)-20-oxo-14,15-dithia-9,19,24-triazatridodecanoic acid di((Z)-non-2-en-1-yl) ester

[0221] 18-((2-(diethylamino)ethyl)carbamoyl)-7,26-dihydroxy-9,24-bis(2-hydroxy-8-(((Z)-non-2-en-1-yl)oxy)-8-oxooctyl)-20-oxo-14,15-dithia-9,19,24-triazatridodecanoic acid di((Z)-non-2-en-1-yl) ester

[0222] 18-((3-(dimethylamino)propyl)carbamoyl)-26-hydroxy-24-(2-hydroxydodecyl)-9-(8-(((Z)-non-2-en-1-yl)oxy)-8-oxooctyl)-20-oxo-14,15-dithia-9,19,24-triazahexadecanoic acid (Z)-non-2-en-1-yl ester

[0223] 18-((2-(diethylamino)ethyl)carbamoyl)-26-hydroxy-24-(2-hydroxydodecyl)-9-(8-(((Z)-non-2-en-1-yl)oxy)-8-oxooctyl)-20-oxo-14,15-dithia-9,19,24-triazahexadecanoic acid (Z)-non-2-en-1-yl ester

[0224] 18-((3-(diethylamino)propyl)carbamoyl)-26-hydroxy-24-(2-hydroxydodecyl)-9-(8-(((Z)-non-2-en-1-yl)oxy)-8-oxooctyl)-20-oxo-14,15-dithia-9,19,24-triazahexadecanoic acid (Z)-non-2-en-1-yl ester

[0225] 18-((2-(dimethylamino)ethyl)carbamoyl)-26-hydroxy-24-(2-hydroxydodecyl)-9-(8-(((Z)-non-2-en-1-yl)oxy)-8-oxooctyl)-20-oxo-14,15-dithia-9,19,24-triazahexadecanoic acid (Z)-non-2-en-1-yl ester

[0226] 18-((2-(dimethylamino)ethyl)(methyl)carbamoyl)-26-hydroxy-24-(2-hydroxydodecyl)-9-(8-(((Z)-non-2-en-1-yl)oxy)-8-oxooctyl)-20-oxo-14,15-dithia-9,19,24-triazahexadecanoic acid(Z)-non-2-en-1-yl ester

[0227] 18-((2-(diethylamino)ethyl)(methyl)carbamoyl)-26-hydroxy-24-(2-hydroxydodecyl)-9-(8-(((Z)-non-2-en-1-yl)oxy)-8-oxooctyl)-20-oxo-14,15-dithia-9,19,24-triazahexadecanoic acid (Z)-non-2-en-1-yl ester

[0228] 18-((3-(1H-imidazol-1-yl)propyl)carbamoyl)-26-hydroxy-24-(2-hydroxydodecyl)-9-(8-(((Z)-non-2-en-1-yl)oxy)-8-oxooctyl)-20-oxo-14,15-dithia-9,19,24-triazahexadecanoic acid(Z)-non-2-en-1-yl ester

[0229] 26-Hydroxy-24-(2-Hydroxydodecyl)-9-(8-(((Z)-Non-2-en-1-yl)oxy)-8-oxooctyl)-20-oxo-18-((2-(piperidin-1-yl)ethyl)carbamoyl)-14,15-dithia-9,19,24-triazahexahexadecanoic acid (Z)-non-2-en-1-yl ester

[0230] 26-Hydroxy-24-(2-Hydroxydodecyl)-18-((4-morpholinobutyl)carbamoyl)-9-(8-(((Z)-non-2-en-1-yl)oxy)-8-oxooctyl)-20-oxo-14,15-dithia-9,19,24-triazahexadecanoic acid (Z)-non-2-en-1-yl ester

[0231] 26-Hydroxy-24-(2-Hydroxydodecyl)-9-(8-(((Z)-Non-2-en-1-yl)oxy)-8-oxooctyl)-20-oxo-18-((4-(pyrrolidone-1-yl)butyl)carbamoyl)-14,15-dithia-9,19,24-triazahexahexadecanoic acid (Z)-non-2-en-1-yl ester

[0232] 5-(bis(2-hydroxydecyl)amino)-N-(19-hydroxy-17-(2-hydroxydodecyl)-2-methyl-7-oxo-11,12-dithia-2,6,17-triazanonadecan-8-yl)pentanamide

[0233] 5-(bis(2-hydroxydecyl)amino)-N-(3-ethyl-19-hydroxy-17-(2-hydroxydodecyl)-7-oxo-11,12-dithia-3,6,17-triazanonadecan-8-yl)pentanamide

[0234] 5-(bis(2-hydroxydecyl)amino)-N-(3-ethyl-20-hydroxy-18-(2-hydroxydodecyl)-8-oxo-12,13-dithia-3,7,18-triazatriacontane-9-yl)pentanamide

[0235] 5-(bis(2-hydroxydecyl)amino)-N-(18-hydroxy-16-(2-hydroxydodecyl)-2-methyl-6-oxo-10,11-dithia-2,5,16-triazaoctacosane-7-yl)pentanamide

[0236] 5-(bis(2-hydroxydecyl)amino)-N-(18-hydroxy-16-(2-hydroxydodecyl)-2,5-dimethyl-6-oxo-10,11-dithia-2,5,16-triazaoctacosane-7-yl)pentanamide

[0237] 5-(bis(2-hydroxydecyl)amino)-N-(3-ethyl-19-hydroxy-17-(2-hydroxydodecyl)-6-methyl-7-oxo-11,12-dithia-3,6,17-triazanonadecan-8-yl)pentanamide

[0238] N-(1-((3-(1H-imidazol-1-yl)propyl)amino)-4-((4-(bis(2-hydroxydodecyl)amino)butyl)dithioalkyl)-1-oxobutyl-2-yl)-5-(bis(2-hydroxydecyl)amino)pentanamide)

[0239] 5-(bis(2-hydroxydecyl)amino)-N-(4-((4-(bis(2-hydroxydodecyl)amino)butyl)dithioalkyl)-1-oxo-1-((3-(piperidin-1-yl)propyl)amino)but-2-yl)pentanamide

[0240] 5-(bis(2-hydroxydecyl)amino)-N-(4-((4-(bis(2-hydroxydodecyl)amino)butyl)dithioalkyl)-1-oxo-1-((2-(pyrrolidone-1-yl)ethyl)amino)but-2-yl)pentanamide

[0241] 5-(bis(2-hydroxydecyl)amino)-N-(4-((4-(bis(2-hydroxydodecyl)amino)butyl)dithioalkyl)-1-oxo-1-((4-(pyrrolidone-1-yl)butyl)amino)but-2-yl)pentanamide

[0242] 18-((3-(dimethylamino)propyl)carbamoyl)-25-hydroxy-23-(2-hydroxydecyl)-20-oxo-9-(6-oxo-6-(undecyloxy)hexyl)-14,15-dithia-9,19,23-triazatriacontane-octadecanoate-9-yl ester

[0243] 18-((2-(diethylamino)ethyl)carbamoyl)-25-hydroxy-23-(2-hydroxydecyl)-20-oxo-9-(6-oxo-6-(undecyloxy)hexyl)-14,15-dithia-9,19,23-triazatriacontane-octadecanoate-9-yl ester

[0244] 8-[4-[[3-[3-[bis(2-hydroxydecyl)amino]propionylamino]-4-[3-(diethylamino)propylamino]-4-oxo-butyl]dithioalkyl]butyl-(6-oxo-6-undecoxy-hexyl)amino]octanoic acid 1-octylnonyl ester

[0245] 8-[4-[[3-[3-[bis(2-hydroxydecyl)amino]propionylamino]-4-[2-(dimethylamino)ethylamino]-4-oxo-butyl]dithioalkyl]butyl-(6-oxo-6-undecoxy-hexyl)amino]octanoic acid 1-octylnonyl ester

[0246] 8-[4-[[3-[3-[bis(2-hydroxydecyl)amino]propionylamino]-4-[2-(dimethylamino)ethyl-methyl-amino]-4-oxo-butyl]dithioalkyl]butyl-(6-oxo-6-undecoxy-hexyl)amino]octanoic acid 1-octylnonyl ester

[0247] 8-[4-[[3-[3-[bis(2-hydroxydecyl)amino]propionylamino]-4-[2-(diethylamino)ethyl-methyl-amino]-4-oxo-butyl]dithioalkyl]butyl-(6-oxo-6-undecoxy-hexyl)amino]octanoic acid 1-octylnonyl ester

[0248] 8-[4-[[3-[3-[bis(2-hydroxydecyl)amino]propionylamino]-4-(3-imidazol-1-ylpropylamino)-4-oxo-butyl]dithioalkyl]butyl-(6-oxo-6-undecoxy-hexyl)amino]octanoic acid 1-octylnonyl ester

[0249] 8-[4-[[3-[3-[bis(2-hydroxydecyl)amino]propionylamino]-4-(4-morpholinobutylamino)-4-oxo-butyl]dithioalkyl]butyl-(6-oxo-6-undecoxy-hexyl)amino]octanoic acid 1-octylnonyl ester

[0250] 8-[4-[[3-[3-[bis(2-hydroxydecyl)amino]propionylamino]-4-oxo-4-(4-pyrrolidine-1-ylbutylamino)butyl]dithioalkyl]butyl-(6-oxo-6-undecoxy-hexyl)amino]octanoic acid 1-octylnonyl ester

[0251] 8-[[4-[[3-[4-[bis(2-hydroxytetradecyl)amino]butyldithioalkyl]-1-[3-(dimethylamino)propylcarbamoyl]propyl]amino]-4-oxo-butyl]-[2-hydroxy-8-[(Z)-non-2-enoxy]-8-oxo-octyl]amino]-7-hydroxy-octanoic acid[(Z)-non-2-enyl] ester

[0252] 8-[[4-[[3-[4-[bis(2-hydroxytetradecyl)amino]butyldithioalkyl]-1-[2-(diethylamino)ethylcarbamoyl]propyl]amino]-4-oxo-butyl]-[2-hydroxy-8-[(Z)-non-2-enoxy]-8-oxo-octyl]amino]-7-hydroxy-octanoic acid[(Z)-non-2-enyl] ester

[0253] 8-[[4-[[3-[4-[bis(2-hydroxytetradecyl)amino]butyldithioalkyl]-1-[3-(diethylamino)propylcarbamoyl]propyl]amino]-4-oxo-butyl]-[2-hydroxy-8-[(Z)-non-2-enoxy]-8-oxo-octyl]amino]-7-hydroxy-octanoic acid[(Z)-non-2-enyl] ester

[0254] 8-[[4-[[3-[4-[bis(2-hydroxytetradecyl)amino]butyldithioalkyl]-1-[2-(dimethylamino)ethylcarbamoyl]propyl]amino]-4-oxo-butyl]-[2-hydroxy-8-[(Z)-non-2-enoxy]-8-oxo-octyl]amino]-7-hydroxy-octanoic acid[(Z)-non-2-enyl] ester

[0255] 8-[[4-[[3-[4-[bis(2-hydroxytetradecyl)amino]butyldithioalkyl]-1-[2-(dimethylamino)ethyl-methyl-carbamoyl]propyl]amino]-4-oxo-butyl]-[2-hydroxy-8-[(Z)-non-2-enoxy]-8-oxo-octyl]amino]-7-hydroxy-octanoic acid[(Z)-non-2-enyl] ester

[0256] 8-[[4-[[3-[4-[bis(2-hydroxytetradecyl)amino]butyldithioalkyl]-1-[2-(diethylamino)ethyl-methyl-carbamoyl]propyl]amino]-4-oxo-butyl]-[2-hydroxy-8-[(Z)-non-2-enoxy]-8-oxo-octyl]amino]-7-hydroxy-octanoic acid[(Z)-non-2-enyl] ester

[0257] 8-[[4-[[3-[4-[bis(2-hydroxytetradecyl)amino]butyldithioalkyl]-1-(3-imidazol-1-ylpropylcarbamoyl)propyl]amino]-4-oxo-butyl]-[2-hydroxy-8-[(Z)-non-2-enoxy]-8-oxo-octyl]amino]-7-hydroxy-octanoic acid[(Z)-non-2-enyl] ester

[0258] 8-[[4-[[3-[4-[bis(2-hydroxytetradecyl)amino]butyldithioalkyl]-1-[2-(1-piperidinyl)ethylcarbamoyl]propyl]amino]-4-oxo-butyl]-[2-hydroxy-8-[(Z)-non-2-enoxy]-8-oxo-octyl]amino]-7-hydroxy-octanoic acid[(Z)-non-2-enyl] ester

[0259] 8-[[4-[[3-[4-[bis(2-hydroxytetradecyl)amino]butyldithioalkyl]-1-(4-morpholinobutylcarbamoyl)propyl]amino]-4-oxo-butyl]-[2-hydroxy-8-[(Z)-non-2-enoxy]-8-oxo-octyl]amino]-7-hydroxy-octanoic acid[(Z)-non-2-enyl] ester

[0260] 8-[[4-[[3-[4-[bis(2-hydroxytetradecyl)amino]butyldithioalkyl]-1-(4-pyrrolidine-1-ylbutylcarbamoyl)propyl]amino]-4-oxo-butyl]-[2-hydroxy-8-[(Z)-non-2-enoxy]-8-oxo-octyl]amino]-7-hydroxy-octanoic acid[(Z)-non-2-enyl] ester

[0261] 8-[[4-[[1-[3-(dimethylamino)propylcarbamoyl]-3-[4-[[8-(1-octylnonoxy)-8-oxo-octyl]-(6-oxo-6-undecoxy-hexyl)amino]butyldithioyl]propyl]amino]-4-oxo-butyl]-[2-hydroxy-8-[(Z)-non-1-enoxy]-8-oxo-octyl]amino]-7-hydroxy-octanoic acid[(Z)-non-1-enyl] ester

[0262] 8-[[4-[[1-[2-(diethylamino)ethylcarbamoyl]-3-[4-[[8-(1-octylnonoxy)-8-oxo-octyl]-(6-oxo-6-undecoxy-hexyl)amino]butyldithioyl]propyl]amino]-4-oxo-butyl]-[2-hydroxy-8-[(Z)-non-2-enoxy]-8-oxo-octyl]amino]-7-hydroxy-octanoic acid[(Z)-non-2-enyl] ester]

[0263] 8-[[4-[[1-[3-(diethylamino)propylcarbamoyl]-3-[4-[[8-(1-octylnonoxy)-8-oxo-octyl]-(6-oxo-6-undecoxy-hexyl)amino]butyldithioyl]propyl]amino]-4-oxo-butyl]-[2-hydroxy-8-[(Z)-non-1-enoxy]-8-oxo-octyl]amino]-7-hydroxy-octanoic acid[(Z)-non-1-enyl] ester]

[0264] 18-((2-(dimethylamino)ethyl)carbamoyl)-26-hydroxy-24-(2-hydroxy-8-(((Z)-non-2-en-1-yl)oxy)-8-oxooctyl)-20-oxo-9-(6-oxo-6-(undecyloxy)hexyl)-14,15-dithia-9,19,24-triazatridodecanoic acid 1-(heptadecano-9-yl)32-((Z)-non-2-en-1-yl) ester

[0265] 8-[[4-[[1-[2-(dimethylamino)ethyl-methyl-carbamoyl]-3-[4-[[8-(1-octylnonoxy)-8-oxo-octyl]-(6-oxo-6-undecoxy-hexyl)amino]butyldithioyl]propyl]amino]-4-oxo-butyl]-[2-hydroxy-8-[(Z)-non-2-enoxy]-8-oxo-octyl]amino]-7-hydroxy-octanoic acid[(Z)-non-2-enyl] ester]

[0266] 8-[[4-[[1-[2-(diethylamino)ethyl-methyl-carbamoyl]-3-[4-[[8-(1-octylnonoxy)-8-oxo-octyl]-(6-oxo-6-undecoxy-hexyl)amino]butyldithioyl]propyl]amino]-4-oxo-butyl]-[2-hydroxy-8-[(Z)-non-2-enoxy]-8-oxo-octyl]amino]-7-hydroxy-octanoic acid[(Z)-non-2-enyl] ester]

[0267] 7-Hydroxy-8-[[2-Hydroxy-8-[(Z)-Non-2-enoxy]-8-oxo-octyl]-[4-[[1-(3-imidazol-1-ylpropylcarbamoyl)-3-[4-[[8-(1-octylnonoxy)-8-oxo-octyl]-(6-oxo-6-undecoxy-hexyl)amino]butyldithioyl]propyl]amino]-4-oxo-butyl]amino]octanoic acid[(Z)-non-2-enyl] ester

[0268] 7-Hydroxy-8-[[2-Hydroxy-8-[(Z)-Non-2-enoxy]-8-oxo-octyl]-[4-[[3-[4-[[8-(1-octylnonoxy)-8-oxo-octyl]-(6-oxo-6-undecoxy-hexyl)amino]butyldithioyl]-1-[2-(1-piperidinyl)ethylcarbamoyl]propyl]amino]-4-oxo-butyl]amino]octanoic acid[(Z)-non-2-enyl] ester

[0269] 7-Hydroxy-8-[[2-Hydroxy-8-[(Z)-Non-2-enoxy]-8-oxo-octyl]-[4-[[1-(4-morpholinobutylcarbamoyl)-3-[4-[[8-(1-octylnonoxy)-8-oxo-octyl]-(6-oxo-6-undecoxy-hexyl)amino]butyldithioyl]propyl]amino]-4-oxo-butyl]amino]octanoic acid[(Z)-non-2-enyl] ester

[0270] 7-Hydroxy-8-[[2-Hydroxy-8-[(Z)-Non-2-enoxy]-8-oxo-octyl]-[4-[[3-[4-[[8-(1-octylnonoxy)-8-oxo-octyl]-(6-oxo-6-undecoxy-hexyl)amino]butyldithioalkyl]-1-(4-pyrrolidine-1-ylbutylcarbamoyl)propyl]amino]-4-oxo-butyl]amino]octanoic acid[(Z)-non-2-enyl] ester

[0271] 8-[4-[[3-[4-[bis(2-hydroxydodecyl)amino]butyrylamino]-4-[3-(dimethylamino)propylamino]-4-oxo-butyl]dithioalkyl]butyl-(6-oxo-6-undecoxy-hexyl)amino]octanoic acid 1-octylnonyl ester

[0272] 8-[4-[[3-[4-[bis(2-hydroxydodecyl)amino]butyrylamino]-4-[2-(diethylamino)ethylamino]-4-oxo-butyl]dithioalkyl]butyl-(6-oxo-6-undecoxy-hexyl)amino]octanoic acid 1-octylnonyl ester

[0273] 8-[4-[[3-[4-[bis(2-hydroxydodecyl)amino]butyrylamino]-4-[3-(diethylamino)propylamino]-4-oxo-butyl]dithioalkyl]butyl-(6-oxo-6-undecoxy-hexyl)amino]octanoic acid 1-octylnonyl ester

[0274] 8-[4-[[3-[4-[bis(2-hydroxydodecyl)amino]butyrylamino]-4-[2-(dimethylamino)ethylamino]-4-oxo-butyl]dithioalkyl]butyl-(6-oxo-6-undecoxy-hexyl)amino]octanoic acid 1-octylnonyl ester

[0275] 8-[4-[[3-[4-[bis(2-hydroxydodecyl)amino]butyrylamino]-4-[2-(dimethylamino)ethyl-methyl-amino]-4-oxo-butyl]dithioalkyl]butyl-(6-oxo-6-undecoxy-hexyl)amino]octanoic acid 1-octylnonyl ester

[0276] 8-[4-[[3-[4-[bis(2-hydroxydodecyl)amino]butyrylamino]-4-[2-(diethylamino)ethyl-methyl-amino]-4-oxo-butyl]dithioalkyl]butyl-(6-oxo-6-undecoxy-hexyl)amino]octanoic acid 1-octylnonyl ester

[0277] 8-[4-[[3-[4-[bis(2-hydroxydodecyl)amino]butyrylamino]-4-(3-imidazol-1-ylpropylamino)-4-oxo-butyl]dithioalkyl]butyl-(6-oxo-6-undecoxy-hexyl)amino]octanoic acid 1-octylnonyl ester

[0278] 8-[4-[[3-[4-[bis(2-hydroxydodecyl)amino]butyrylamino]-4-oxo-4-[2-(1-piperidinyl)ethylamino]butyl]dithioalkyl]butyl-(6-oxo-6-undecoxy-hexyl)amino]octanoic acid 1-octylnonyl ester

[0279] 8-[4-[[3-[4-[bis(2-hydroxydodecyl)amino]butyrylamino]-4-(4-morpholinobutylamino)-4-oxo-butyl]dithioalkyl]butyl-(6-oxo-6-undecoxy-hexyl)amino]octanoic acid 1-octylnonyl ester

[0280] 8-[4-[[3-[4-[bis(2-hydroxydodecyl)amino]butyrylamino]-4-oxo-4-(4-pyrrolidine-1-ylbutylamino)butyl]dithioalkyl]butyl-(6-oxo-6-undecoxy-hexyl)amino]octanoic acid 1-octylnonyl ester

[0281] 18-((3-(2,7a-dihydro-1H-benzo[d]imidazol-1-yl)propyl)carbamoyl)-27-hydroxy-25-(2-hydroxydecyl)-20-oxo-9-(6-oxo-6-(undecyloxy)hexyl)-14,15-dithia-9,19,25-triazapentadecanoic acid heptadecano-9-yl ester

[0282] 18-((3-(1H-imidazol-1-yl)propyl)carbamoyl)-27-hydroxy-25-(2-hydroxydecyl)-20-oxo-9-(6-oxo-6-(undecyloxy)hexyl)-14,15-dithia-9,19,25-triazapentadecanoic acid heptadecano-9-yl ester

[0283] N-(3-(1H-imidazol-1-yl)propyl)-4-((4-(bis(2-hydroxydecyl)amino)butyl)dithio)-2-(4-(bis(2-hydroxydodecyl)amino)butyramido)butyramide

[0284] N-(1-((4-(1H-imidazol-1-yl)butyl)amino)-4-((4-(bis(2-hydroxytetradecyl)amino)butyl)dithioalkyl)-1-oxobutyl-2-yl)-5-(bis(2-hydroxydecyl)amino)pentanamide

[0285] N-(1-((2-(1H-imidazol-1-yl)ethyl)amino)-4-((4-(bis(2-hydroxytetradecyl)amino)butyl)dithioalkyl)-1-oxobutyl-2-yl)-5-(bis(2-hydroxydecyl)amino)pentanamide

[0286] 5-(bis(2-hydroxydecyl)amino)-N-(4-((4-(bis(2-hydroxytetradecyl)amino)butyl)dithioalkyl)-1-((3-(2-methyl-1H-imidazol-1-yl)propyl)amino)-1-oxobut-2-yl)pentanamide

[0287] N-(1-((2-(1H-indol-3-yl)ethyl)amino)-4-((4-(bis(2-hydroxytetradecyl)amino)butyl)dithioalkyl)-1-oxobutyl-2-yl)-5-(bis(2-hydroxydecyl)amino)pentanamide)

[0288] 2-[4-[bis(2-hydroxydodecyl)amino]butyrylamino]-4-[4-[bis[(Z)-2-hydroxyoctadec-9-enyl]amino]butyldithioalkyl]-N-[3-(dimethylamino)propyl]butyramide

[0289] 2-[4-[bis(2-hydroxydodecyl)amino]butyrylamino]-4-[4-[bis[(Z)-2-hydroxyoctadec-9-enyl]amino]butyldithioalkyl]-N-[2-(diethylamino)ethyl]butyramide

[0290] 2-[4-[bis(2-hydroxydodecyl)amino]butyrylamino]-4-[4-[bis[(Z)-2-hydroxyoctadec-9-enyl]amino]butyldithioalkyl]-N-[3-(diethylamino)propyl]butyramide

[0291] 2-[4-[bis(2-hydroxydodecyl)amino]butyrylamino]-4-[4-[bis[(Z)-2-hydroxyoctadec-9-enyl]amino]butyldithioalkyl]-N-[2-(dimethylamino)ethyl]butyramide

[0292] 2-[4-[bis(2-hydroxydodecyl)amino]butyrylamino]-4-[4-[bis[(Z)-2-hydroxyoctadec-9-enyl]amino]butyldithioalkyl]-N-[2-(dimethylamino)ethyl]-N-methylbutyramide; methane

[0293] 2-[4-[bis(2-hydroxydodecyl)amino]butyrylamino]-4-[4-[bis[(Z)-2-hydroxyoctadec-9-enyl]amino]butyldithioalkyl]-N-[2-(diethylamino)ethyl]-N-methylbutyramide

[0294] N-(3-(1H-imidazol-1-yl)propyl)-4-((4-(bis((Z)-2-hydroxyoctadec-9-en-1-yl)amino)butyl)dithioyl)-2-(4-(bis(2-hydroxydodecyl)amino)butyramido)butyramide

[0295] 2-[4-[bis(2-hydroxydodecyl)amino]butyrylamino]-4-[4-[bis[(Z)-2-hydroxyoctadec-9-enyl]amino]butyldithioalkyl]-N-[3-(2-methylimidazol-1-yl)propyl]butyramide

[0296] 2-[4-[bis(2-hydroxydodecyl)amino]butyrylamino]-4-[4-[bis[(Z)-2-hydroxyoctadec-9-enyl]amino]butyldithioalkyl]-N-(4-morpholinobutyl)butyramide

[0297] 2-[4-[bis(2-hydroxydodecyl)amino]butyrylamino]-4-[4-[bis[(Z)-2-hydroxyoctadec-9-enyl]amino]butyldithioalkyl]-N-(4-pyrrolidine-1-ylbutyl)butyramide

[0298] 4-((4-(bis(2-hydroxydecyl)amino)butyl)dithioalkyl)-2-(4-(bis(2-hydroxydodecyl)amino)butamido)-N-(3-(dimethylamino)propyl)butamido)

[0299] 4-((4-(bis(2-hydroxydecyl)amino)butyl)dithioalkyl)-2-(4-(bis(2-hydroxydodecyl)amino)butamido)-N-(2-(diethylamino)ethyl)butamido)

[0300] 4-((4-(bis(2-hydroxydecyl)amino)butyl)dithioalkyl)-2-(4-(bis(2-hydroxydodecyl)amino)butamido)-N-(3-(diethylamino)propyl)butamido)

[0301] 4-((4-(bis(2-hydroxydecyl)amino)butyl)dithioalkyl)-2-(4-(bis(2-hydroxydodecyl)amino)butamido)-N-(2-(dimethylamino)ethyl)butamido)

[0302] 4-((4-(bis(2-hydroxydecyl)amino)butyl)dithioalkyl)-2-(4-(bis(2-hydroxydodecyl)amino)butamido)-N-(2-(dimethylamino)ethyl)-N-methylbutamido)

[0303] 4-((4-(bis(2-hydroxydecyl)amino)butyl)dithioalkyl)-2-(4-(bis(2-hydroxydodecyl)amino)butamido)-N-(2-(diethylamino)ethyl)-N-methylbutamido)

[0304] 4-((4-(bis(2-hydroxydecyl)amino)butyl)dithioalkyl)-2-(4-(bis(2-hydroxydodecyl)amino)butamido)-N-(3-(2-methyl-1H-imidazol-1-yl)propyl)butamido

[0305] 4-((4-(bis(2-hydroxydecyl)amino)butyl)dithioalkyl)-2-(4-(bis(2-hydroxydodecyl)amino)butamido)-N-(4-morpholinobutyl)butamido)

[0306] 4-((4-(bis(2-hydroxydecyl)amino)butyl)dithioalkyl)-2-(4-(bis(2-hydroxydodecyl)amino)butamido)-N-(4-(pyrrolidone-1-yl)butyl)butamido)

[0307] 2-(4-(bis(2-hydroxydodecyl)amino)butamido)-4-((4-(bis(2-hydroxydodecyl)amino)butyl)dithio)-N-(3-(dimethylamino)propyl)butamido)

[0308] 2-(4-(bis(2-hydroxydodecyl)amino)butyramido)-4-((4-(bis(2-hydroxydodecyl)amino)butyl)dithio)-N-(2-(diethylamino)ethyl)butyramido)

[0309] 2-(4-(bis(2-hydroxydodecyl)amino)butyramido)-4-((4-(bis(2-hydroxydodecyl)amino)butyl)dithioalkyl)-N-(3-(diethylamino)propyl)butyramido

[0310] 2-(4-(bis(2-hydroxydodecyl)amino)butyramido)-4-((4-(bis(2-hydroxydodecyl)amino)butyl)dithio)-N-(2-(dimethylamino)ethyl)butyramido

[0311] 2-(4-(bis(2-hydroxydodecyl)amino)butamido)-4-((4-(bis(2-hydroxydodecyl)amino)butyl)dithio)-N-(2-(dimethylamino)ethyl)-N-methylbutamido)

[0312] 2-(4-(bis(2-hydroxydodecyl)amino)butamido)-4-((4-(bis(2-hydroxydodecyl)amino)butyl)dithio)-N-(2-(diethylamino)ethyl)-N-methylbutamido)

[0313] 2-(4-(bis(2-hydroxydodecyl)amino)butamido)-4-((4-(bis(2-hydroxydodecyl)amino)butyl)dithio)-N-(3-(4-(2-hydroxyethyl)piperazin-1-yl)propyl)butamido)

[0314] 2-(4-(bis(2-hydroxydodecyl)amino)butamido)-4-((4-(bis(2-hydroxydodecyl)amino)butyl)dithio)-N-(4-morpholinobutyl)butamido)

[0315] 2-(4-(bis(2-hydroxydodecyl)amino)butamido)-4-((4-(bis(2-hydroxydodecyl)amino)butyl)dithioalkyl)-N-(4-(pyrrolidone-1-yl)butyl)butamido)

[0316] And its pharmaceutically acceptable salts and esters.

[0317] lipid synthesis

[0318] The lipids described herein, namely homocysteine-based lipids having formula (I), such as compounds having formulas (IA)-(II), can be prepared according to methods known in the art.

[0319] More specifically, the present invention provides a first method for preparing a compound having formula (I) as defined herein, the method comprising the step of: taking a compound having formula (II):

[0320] (II)

[0321] Reaction with compounds having formula (III):

[0322] (III)

[0323] Where m, n, p, R 1 R 2 R 3 R 4 R 5 R 6 and R 7 As defined in this article.

[0324] In the examples, the reaction can be carried out in an organic solvent, preferably a nonpolar solvent such as chloroform. Compound (III) can be used advantageously in excess relative to compounds having formula (II).

[0325] Typically, the reaction can be carried out over a wide temperature range from 0°C to the reflux temperature of the reaction mixture, preferably at room temperature.

[0326] In an embodiment, the method further comprises preparing a compound having formula (II) by: making a compound having formula (IV)

[0327] (IV)

[0328] and

[0329] Compounds having formula (V):

[0330] (V)

[0331] Reaction with compounds having formula (VI)

[0332] (VI)

[0333] Where m, n, and R 3 R 4 R 5 R 6 and R7 As defined in this article.

[0334] In the examples, the reaction can be carried out in an organic solvent, preferably a nonpolar solvent such as chloroform.

[0335] Typically, the reaction can be carried out over a wide temperature range from 0°C to the reflux temperature of the reaction mixture, typically at room temperature.

[0336] The present invention also provides an alternative method for preparing compounds having formula (I), the method comprising the following one-pot step: preparing a compound having formula (III):

[0337] (III)

[0338] Compounds having formula (IV)

[0339] (IV)

[0340] Compounds having formula (V):

[0341] (V)

[0342] Reaction with compounds having formula (VI):

[0343] (VI)

[0344] Where m, n, p, R 1 R 2 R 3 R 4 R 5 R 6 and R 7 As defined in this article.

[0345] In the examples, the reaction can be carried out in an organic solvent over a wide temperature range from 0°C to the reflux temperature of the reaction mixture.

[0346] In two alternative embodiments, the method may further include the step of preparing a compound having formula (IV) by: in the presence of a coupling agent, causing a compound having formula (VII) to be prepared.

[0347] (VII)

[0348] Where X represents an H or hydroxyl protecting group, and R 6 and R 7 As defined in equation (I) in this paper;

[0349] Reaction with compounds having formula (VIII)

[0350] (VIII)

[0351] Hal represents a halogen atom.

[0352] Optionally, the obtained product is then deprotected in an acidic medium, wherein X is a hydroxyl protecting group.

[0353] The reaction of compound (VII) with compound (VIII) typically produces an adduct with the following formula:

[0354] HHal

[0355] Where X represents an H or hydroxyl protecting group, and R 6 and R 7 As defined in equation (I).

[0356] The reaction can be carried out in a polar aprotic solvent (such as dichloromethane (DCM)) in the presence of a nucleophilic catalyst (such as 4-dimethylaminopyridine (DMAP)) and a carboxyl activator (such as 1-ethyl-3-(3-dimethylaminopropyl)carbodiimide (EDC)).

[0357] In the embodiments, the deprotection step may be performed in one pot on the adducts generated by the reaction of compounds (VII) and (VIII), or after the adducts have been isolated and purified.

[0358] In an embodiment, the deprotection can be carried out by reacting the obtained product with an acid, such as hydrogen fluoride, typically in the form of a 70% HF-pyridine complex. This reaction can be carried out in an organic solvent, preferably a polar aprotic solvent such as tetrahydrofuran (THF). Typically, the reaction can be carried out over a wide temperature range from 0°C to the reflux temperature of the reaction mixture.

[0359] Compounds having formula (III) can be prepared by: typically in a polar aprotic solvent such as dichloromethane (DCM), making the compound having formula (III)... compounds

[0360] Tri represents the triphenylmethyl group, which reacts with acids such as trifluoroacetic acid (TFA) and triethylsilane.

[0361] Particles

[0362] Lipids having formula (I) as described herein can be used to prepare compositions that can be used to deliver nucleic acids.

[0363] In the examples, the composition comprises mRNA encoding an antigen (e.g., an antigen from an infectious agent).

[0364] In some embodiments, the composition is a suitable delivery medium. In an example, the composition is an LPN, such as a liposome delivery medium.

[0365] The terms “liposome delivery medium” and “liposome composition” are used interchangeably.

[0366] In another aspect, the present invention is characterized by a lipid nanoparticle (LNP), such as a liposome, the LNP comprising a compound having formula (I) according to the present invention, and optionally encapsulating nucleic acids.

[0367] In the embodiments, the nucleic acid is mRNA encoding a peptide or protein.

[0368] In embodiments, the LNP may further comprise one or more components selected from: stealth lipids (e.g., PEGylated lipids); structural lipids (e.g., cholesterol-based lipids); and accessory lipids.

[0369] In the embodiments, the composition or LNP may further comprise one or more lipids selected from the group consisting of one or more cationic lipids, one or more non-cationic lipids, and one or more PEG-modified lipids.

[0370] In some embodiments, lipids having formula (I) as described herein, as well as pharmaceutical and liposome compositions comprising such lipids, can be used in formulations to facilitate the delivery of encapsulated material (e.g., one or more polynucleotides such as mRNA) to one or more target cells and subsequently transfect one or more target cells. For example, in some embodiments, the lipids (and compositions, such as liposome compositions comprising such lipids) described herein are characterized by causing one or more of the following: receptor-mediated endocytosis, clathrin-mediated and pit-mediated endocytosis, phagocytosis and macropinocytosis, fusogenicity, endosome or lysosomal disruption, and / or releasability, which provides advantages of such compounds over other similarly classified lipids.

[0371] According to the present invention, nucleic acids, such as mRNA encoding proteins (e.g., full-length, fragments, or portions of proteins), as described herein, can be delivered via a delivery medium comprising lipids having formula (I) as described herein.

[0372] As used herein, the terms “delivery medium,” “transfer medium,” “nanoparticle,” or grammatical equivalents are used interchangeably.

[0373] For example, the present invention provides a composition (e.g., a pharmaceutical composition) comprising a lipid having formula (I) as described herein and one or more polynucleotides. The composition (e.g., a pharmaceutical composition) may further 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.

[0374] In some embodiments, the composition exhibits an enhanced (e.g., increased) ability to transfect 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 a lipid and / or pharmaceutical composition disclosed herein (e.g., an LPN formulation comprising a lipid having formula (I) as described herein, encapsulating one or more polynucleotides), such that the one or more target cells are transfected by the encapsulated material (e.g., one or more polynucleotides). As used herein, the term “transfect” or “transfection” refers to the introduction of one or more encapsulated materials (e.g., nucleic acids and / or polynucleotides) into cells, or preferably into target cells. The introduced polynucleotides may be stably or transiently maintained in the target cells. The term “transfection efficiency” refers to the relative amount of such encapsulated material (e.g., polynucleotides) absorbed, introduced into, and / or expressed by the transfected target cells. In practice, transfection efficiency can be estimated by the amount of reporter polynucleotide product 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 the encapsulated material (e.g., one or more polynucleotides) will be delivered to the pathological site and subsequently expressed, while minimizing potential systemic adverse reactions or toxicities associated with the compound or its encapsulated contents.

[0375] Transfection of one or more target cells with polynucleotides, for example, encapsulated in one or more lipid nanoparticles containing the pharmaceutical or liposome compositions disclosed herein, can preferably stimulate the production of products (e.g., peptides or proteins) encoded by such polynucleotides and enhance the ability of such target cells to express polynucleotides and produce, for example, the target peptide or protein. 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 mRNA.

[0376] Furthermore, 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, the 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 of the compounds or pharmaceutical 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 producing (and in some cases excreted) 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 target proteins.

[0377] One or more liposome compositions enriched with the lipids disclosed herein can be used as a means to improve (e.g., reduce) the toxicity of such enriched liposome compositions or otherwise impart one or more desired properties to such enriched liposome compositions (e.g., improved delivery of encapsulated polynucleotides to one or more target cells and / or reduced in vivo toxicity of the liposome compositions). Therefore, pharmaceutical compositions comprising one or more of the lipids disclosed herein, particularly liposome compositions, are also contemplated.

[0378] Therefore, in some embodiments, the compounds described herein (e.g., lipids having formula (I) as described herein) are lipids that can be used as components of liposome compositions to promote 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 permeation or fusion with the lipid membrane of such target cells).

[0379] As used herein, liposome delivery mediators (e.g., lipid nanoparticles) are typically characterized as microvesicles with an internal aqueous space isolated from an external medium by a membrane having one or more bilayers. The bilayer membrane of a liposome is typically formed of amphiphilic molecules, such as synthetic or naturally derived lipids containing spatially separated hydrophilic and hydrophobic domains (Lasic, Trends Biotechnol. 16: 307-321, 1998). The bilayer membrane of a liposome can also be formed of amphiphilic polymers and surfactants (e.g., polymers, nonionic surfactant vesicles, etc.). In the context of this invention, liposome delivery mediators are typically used to transport desired mRNA to target cells or tissues.

[0380] In some embodiments, such compositions (e.g., liposome compositions) are loaded with or otherwise encapsulated with materials such as, for example, one or more biologically active polynucleotides (e.g., mRNA).

[0381] 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, and at least one lipid is a lipid as described herein (e.g., a lipid having formula (I) 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 an antigen from influenza virus or respiratory syncytial virus (RSV).

[0382] In the embodiments, the composition (e.g., a pharmaceutical composition) comprises nucleic acid encapsulated in liposomes, wherein the liposomes comprise lipids having formula (I) as described herein.

[0383] In the embodiments, the liposome delivery medium (e.g., lipid nanoparticles) may have a net positive charge.

[0384] In the embodiments, the liposome delivery medium (e.g., lipid nanoparticles) may have a net negative charge.

[0385] In the embodiments, the liposome delivery medium (e.g., lipid nanoparticles) may have a net neutral charge.

[0386] In the embodiments, the lipid nanoparticles encapsulating nucleic acids (e.g., mRNA encoding peptides or polypeptides) comprise one or more lipids having formula (I) as described herein.

[0387] For example, the amount of lipids having formula (I) in a composition as described herein 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 the liposome composition).

[0388] In the embodiments of the pharmaceutical compositions described herein, lipids having formula (I) 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., liposome composition).

[0389] In embodiments, lipids having formula (I) 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 embodiments, lipids having formula (I) 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 molar amount of all lipids present in the composition (such as a liposome delivery medium).

[0390] In the embodiments, the amount of lipids having formula (I) as described herein is present in an amount 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).

[0391] In the embodiments, the amount of lipids having formula (I) as described herein is present in an amount 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 the total lipids in the composition (e.g., liposome composition).

[0392] In embodiments, 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 lipids having formula (I) as described herein. In embodiments, 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 lipids having formula (I) as described herein. In embodiments, 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 lipids having formula (I) as described herein. In embodiments, this percentage results in improved beneficial effects (e.g., improved delivery to target tissues such as the liver or lungs).

[0393] The amount of lipids having formula (I) in the composition as described herein can also be described as a percentage (“mol%”) of the total molar amount of lipids in the composition (e.g., the total molar amount of all lipids present in the liposome delivery medium).

[0394] In the embodiments of the pharmaceutical compositions described herein, lipids having formula (I) as described herein are present in an amount of about 0.5 mol% to about 30 mol% (e.g., about 0.5 mol% to about 20 mol%) of the combined molar amount of all lipids present in the composition (such as liposome delivery media).

[0395] In embodiments, lipids having formula (I) as described herein are present in an amount of about 0.5 mol% to about 5 mol%, about 1 mol% to about 10 mol%, about 5 mol% to about 20 mol%, or about 10 mol% to about 20 mol% of the combined molar weight of all lipids present in the composition (such as a liposome delivery medium). In embodiments, lipids having formula (I) as described herein are present in an amount of about 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%, or about 5 mol% to about 25 mol% of the combined dry weight of all lipids present in the composition (such as a liposome delivery medium).

[0396] In some embodiments, lipids having formula (I) as described herein may comprise about 0.1 mol% to about 50 mol% of the total lipids in the composition (e.g., a liposome delivery medium), or 0.5 mol% to about 50 mol%, or about 1 mol% to about 25 mol%, or about 1 mol% to about 10 mol%.

[0397] In some embodiments, lipids having formula (I) as described herein may comprise more than about 0.1 mol%, or more than about 0.5 mol%, or more than about 1 mol%, or more than about 5 mol% of the total lipids in the lipid nanoparticles.

[0398] In some embodiments, lipids having formula (I) as described herein may comprise 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).

[0399] In the embodiments, the amount of lipids having formula (I) as described herein is present in an amount 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 dry weight of total lipids in the composition (e.g., a liposome composition).

[0400] In the embodiments, the amount of lipids having formula (I) as described herein is present in an amount 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% of the combined dry weight of the total lipids in the composition (e.g., liposome composition).

[0401] In the embodiments, this percentage leads to improved beneficial effects (e.g., improved delivery to target tissues such as the liver or lungs).

[0402] In the embodiments, the composition further comprises one or more lipids (e.g., one or more lipids selected from the group consisting of one or more cationic lipids, one or more non-cationic lipids and one or more PEG-modified lipids).

[0403] In some embodiments, such pharmaceutical (e.g., liposome) compositions comprise one or more of PEG-modified lipids, non-cationic lipids, and cholesterol lipids. In some 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 some embodiments, such pharmaceutical (e.g., liposome) compositions comprise: one or more PEG-modified lipids and one or more cholesterol lipids.

[0404] In the embodiments, the composition (e.g., lipid nanoparticles) encapsulating nucleic acids (e.g., mRNA encoding peptides or polypeptides) comprises one or more lipids having formula (I) as described herein, and one or more lipids selected from the group consisting of cationic lipids, non-cationic lipids, and PEGylated lipids.

[0405] In the embodiments, the composition encapsulating nucleic acid (e.g., mRNA encoding peptide or polypeptide) (e.g., lipid nanoparticles) comprises one or more lipids having formula (I) as described herein; one or more lipids selected from the group consisting of cationic lipids, non-cationic lipids and PEGylated lipids; and further comprises cholesterol-based lipids.

[0406] In the embodiments, the lipid nanoparticles encapsulating nucleic acids (e.g., mRNA encoding peptides or polypeptides) comprise one or more lipids having formula (I) as described herein, and one or more lipids selected from the group consisting of cationic lipids, non-cationic lipids, PEGylated lipids, and cholesterol-based lipids.

[0407] According to various embodiments, the selection of cationic lipids, non-cationic lipids, and / or PEG-modified lipids constituting the lipid nanoparticles, and the relative molar ratios of such lipids to each other, are based on the characteristics of one or more 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, fusionogenicity, and toxicity of the selected one or more lipids. Therefore, the molar ratios can be adjusted accordingly.

[0408] In addition to any lipid having formula (I) as described, the composition may contain one or more other cationic lipids.

[0409] In some embodiments, LPN 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.

[0410] Suitable additional cationic lipids for use in the compositions include those described in International Patent Publication WO 2010 / 144740, which is incorporated herein by reference. In some embodiments, these compositions comprise the cationic lipid 4-(dimethylamino)butyric acid (6Z,9Z,28Z,31Z)-heptadecane-6,9,28,31-tetraen-19-yl ester having the following compound structure:

[0411]

[0412] And its pharmaceutically acceptable salts.

[0413] Other suitable additional cationic lipids for use in the composition include ionizable cationic lipids as described in International Patent Publication WO2013 / 149140, which is incorporated herein by reference. In some embodiments, these compositions comprise cationic lipids having one of the following formulas:

[0414] ,

[0415] Or a pharmaceutically acceptable salt thereof, wherein R1 and R2 are each independently selected from hydrogen, optionally substituted, variable saturated or unsaturated C1-C 20 Alkyl groups and optionally substituted C6-C groups that are either saturated or unsaturated 20 The group consisting of acyl groups; wherein L1 and L2 are each independently selected from hydrogen, optionally substituted C1-C 30 Alkyl groups, optionally substituted, variable unsaturated C1-C 30 Alkenyl and optionally substituted C1-C 30 The group consisting of alkynyl groups; wherein m and o are each independently selected from the group consisting of zero and any positive integers (e.g., where m is three); and wherein n is zero or any positive integer (e.g., where n is one). In some embodiments, these compositions comprise a cationic lipid (15Z, 18Z)-N,N-dimethyl-6-(9Z, 12Z)-octadec-9,12-dien-1-yl) tetradec-15,18-dien-1-amine (“HGT5000”):

[0416] (HGT-5000)

[0417] And its pharmaceutically acceptable salts. In some embodiments, these compositions comprise a cationic lipid (15Z, 18Z)-N,N-dimethyl-6-((9Z, 12Z)-octadec-9,12-dien-1-yl)tetradec-4,15,18-trien-1-amine (“HGT5001”) having the following compound structure:

[0418] (HGT-5001)

[0419] And its pharmaceutically acceptable salts. In some embodiments, it includes a cationic lipid having the following compound structure and (15Z,18Z)-N,N-dimethyl-6-((9Z,12Z)-octadec-9,12-dien-1-yl)tetradec-5,15,18-trien-1-amine (“HGT5002”):

[0420] (HGT-5002)

[0421] And its pharmaceutically acceptable salts.

[0422] Other suitable additional cationic lipids for use in the compositions include cationic lipids described as amino alcohol lipids in International Patent Publication WO2010 / 053572, which is incorporated herein by reference. In some embodiments, these compositions comprise cationic lipids having the following compound structures:

[0423]

[0424] And its pharmaceutically acceptable salts.

[0425] Other suitable additional cationic lipids for use in the composition include cationic lipids as described in International Patent Publication WO2016 / 118725, which is incorporated herein by reference. In some embodiments, these compositions comprise cationic lipids having the following compound structures:

[0426]

[0427] And its pharmaceutically acceptable salts.

[0428] Other suitable additional cationic lipids for use in the compositions include cationic lipids as described in International Patent Publication WO2016 / 118724, which is incorporated herein by reference. In some embodiments, these compositions comprise cationic lipids having the following compound structures:

[0429]

[0430] And its pharmaceutically acceptable salts.

[0431] Other suitable cationic lipids for use in the composition include cationic lipids having the following formula: 14,25-ditridecyl-15,18,21,24-tetraaza-octacosan, and pharmaceutically acceptable salts thereof.

[0432] Other suitable additional cationic lipids for use in the compositions include cationic lipids as described in International Patent Publications WO2013 / 063468 and WO 2016 / 205691, which are incorporated herein by reference. In some embodiments, these compositions comprise cationic lipids having the following formula:

[0433]

[0434] Or a pharmaceutically acceptable salt thereof, wherein R L Independently for the optionally substituted C6-C in each case 40 Alkenyl groups. In some embodiments, these compositions comprise cationic lipids having the following compound structures:

[0435] (cKK-E12)

[0436] And its pharmaceutically acceptable salts.

[0437] In some embodiments, these compositions comprise cationic lipids having the following compound structures:

[0438] (OF-02)

[0439] And its pharmaceutically acceptable salts.

[0440] In some embodiments, these compositions comprise cationic lipids having the following compound structures:

[0441]

[0442] And its pharmaceutically acceptable salts.

[0443] In some embodiments, these compositions comprise cationic lipids having the following compound structures:

[0444]

[0445] And its pharmaceutically acceptable salts.

[0446] Other suitable additional cationic lipids for use in the composition include cationic lipids as described in International Patent Publication WO2015 / 184256, which is incorporated herein by reference. In some embodiments, these compositions comprise cationic lipids having the following formula:

[0447]

[0448] Or a pharmaceutically acceptable salt thereof, wherein each X is independently O or S; each Y is independently O or S; each m is independently 0 to 20; each n is independently 1 to 6; each R A Independently, it is hydrogen, optionally substituted C1-50 alkyl, optionally substituted C2-50 alkenyl, optionally substituted C2-50 ynyl, optionally substituted C3-10 carbocyclic, optionally substituted 3-14-membered heterocyclic, optionally substituted C6-14 aryl, optionally substituted 5-14-membered heteroaryl, or halogen; and each R B Independently, it is hydrogen, optionally substituted C1-50 alkyl, optionally substituted C2-50 alkenyl, optionally substituted C2-50 alkynyl, optionally substituted C3-10 carbocyclic, optionally substituted 3-14-membered heterocyclic, optionally substituted C6-14 aryl, optionally substituted 5-14-membered heteroaryl, or halogen. In some embodiments, these compositions comprise a cationic lipid “target 23” having the following compound structure:

[0449] (Target 23)

[0450] And its pharmaceutically acceptable salts.

[0451] Other suitable additional cationic lipids for use in the composition include cationic lipids as described in International Patent Publication WO2016 / 004202, which is incorporated herein by reference. In some embodiments, these compositions comprise cationic lipids having the following compound structures:

[0452] ,in

[0453] Or its pharmaceutically acceptable salt.

[0454] In some embodiments, these compositions comprise cationic lipids having the following compound structures:

[0455]

[0456] Or its pharmaceutically acceptable salt.

[0457] In some embodiments, these compositions comprise cationic lipids having the following compound structures:

[0458]

[0459] Or its pharmaceutically acceptable salt.

[0460] Other suitable additional cationic lipids for use in the composition include cationic lipids as described in J. McClellan, MCKing, Cell 2010, 141, 210-217 and Whitehead et al., Nature Communications (2014) 5:4277, which are incorporated herein by reference. In some embodiments, the cationic lipids of these compositions comprise cationic lipids having the following compound structures:

[0461]

[0462] And its pharmaceutically acceptable salts.

[0463] Other suitable additional cationic lipids for use in the compositions include cationic lipids as described in International Patent Publication WO2015 / 199952, which is incorporated herein by reference. In some embodiments, these compositions comprise cationic lipids having the following compound structures:

[0464]

[0465] And its pharmaceutically acceptable salts.

[0466] In some embodiments, these compositions comprise cationic lipids having the following compound structures:

[0467]

[0468] And its pharmaceutically acceptable salts.

[0469] In some embodiments, these compositions comprise cationic lipids having the following compound structures:

[0470]

[0471] And its pharmaceutically acceptable salts.

[0472] In some embodiments, these compositions comprise cationic lipids having the following compound structures:

[0473]

[0474] And its pharmaceutically acceptable salts.

[0475] In some embodiments, these compositions comprise cationic lipids having the following compound structures:

[0476]

[0477] And its pharmaceutically acceptable salts.

[0478] In some embodiments, these compositions comprise cationic lipids having the following compound structures:

[0479]

[0480] And its pharmaceutically acceptable salts.

[0481] In some embodiments, these compositions comprise cationic lipids having the following compound structures:

[0482]

[0483] And its pharmaceutically acceptable salts.

[0484] In some embodiments, these compositions comprise cationic lipids having the following compound structures:

[0485]

[0486] And its pharmaceutically acceptable salts.

[0487] In some embodiments, these compositions comprise cationic lipids having the following compound structures:

[0488]

[0489] And its pharmaceutically acceptable salts.

[0490] In some embodiments, these compositions comprise cationic lipids having the following compound structures:

[0491]

[0492] And its pharmaceutically acceptable salts.

[0493] In some embodiments, these compositions comprise cationic lipids having the following compound structures:

[0494]

[0495] And its pharmaceutically acceptable salts.

[0496] In some embodiments, these compositions comprise cationic lipids having the following compound structures:

[0497]

[0498] And its pharmaceutically acceptable salts.

[0499] In some embodiments, these compositions comprise cationic lipids having the following compound structures:

[0500]

[0501] And its pharmaceutically acceptable salts.

[0502] Other suitable additional cationic lipids for use in the composition include cationic lipids as described in International Patent Publication WO2017 / 004143, which is incorporated herein by reference.

[0503] In some embodiments, these compositions comprise cationic lipids having the following compound structures:

[0504]

[0505] And its pharmaceutically acceptable salts.

[0506] In some embodiments, these compositions comprise cationic lipids having the following compound structures:

[0507]

[0508] And its pharmaceutically acceptable salts.

[0509] In some embodiments, these compositions comprise cationic lipids having the following compound structures:

[0510]

[0511] And its pharmaceutically acceptable salts.

[0512] In some embodiments, these compositions comprise cationic lipids having the following compound structures:

[0513]

[0514] And its pharmaceutically acceptable salts.

[0515] In some embodiments, these compositions comprise cationic lipids having the following compound structures:

[0516]

[0517] And its pharmaceutically acceptable salts.

[0518] In some embodiments, these compositions comprise cationic lipids having the following compound structures:

[0519]

[0520] And its pharmaceutically acceptable salts.

[0521] In some embodiments, these compositions comprise cationic lipids having the following compound structures:

[0522]

[0523] And its pharmaceutically acceptable salts.

[0524] In some embodiments, these compositions comprise cationic lipids having the following compound structures:

[0525]

[0526] And its pharmaceutically acceptable salts.

[0527] In some embodiments, these compositions comprise cationic lipids having the following compound structures:

[0528]

[0529] And its pharmaceutically acceptable salts.

[0530] In some embodiments, these compositions comprise cationic lipids having the following compound structures:

[0531]

[0532] And its pharmaceutically acceptable salts.

[0533] In some embodiments, these compositions comprise cationic lipids having the following compound structures:

[0534]

[0535] And its pharmaceutically acceptable salts.

[0536] In some embodiments, these compositions comprise cationic lipids having the following compound structures:

[0537]

[0538] And its pharmaceutically acceptable salts.

[0539] In some embodiments, these compositions comprise cationic lipids having the following compound structures:

[0540]

[0541] And its pharmaceutically acceptable salts.

[0542] In some embodiments, these compositions comprise cationic lipids having the following compound structures:

[0543]

[0544] And its pharmaceutically acceptable salts.

[0545] In some embodiments, these compositions comprise cationic lipids having the following compound structures:

[0546]

[0547] And its pharmaceutically acceptable salts.

[0548] In some embodiments, these compositions comprise cationic lipids having the following compound structures:

[0549]

[0550] And its pharmaceutically acceptable salts.

[0551] In some embodiments, these compositions comprise cationic lipids having the following compound structures:

[0552]

[0553] And its pharmaceutically acceptable salts.

[0554] Other suitable additional cationic lipids for use in the composition include cationic lipids as described in International Patent Publication WO2017 / 075531, which is incorporated herein by reference. In some embodiments, these compositions comprise cationic lipids having the following formula:

[0555]

[0556] Or its pharmaceutically acceptable salt, wherein L 1 or L 2 One of them is -O(C=O)-, -(C=O)O-, -C(=O)-, -O-, -S(O) x , -SS-, -C(=O)S-, -SC(=O)-, -NR a C(=O)-、-C(=O)NR a -、NR a C(=O)NR a -、-OC(=O)NR a -or-NR aC(=O)O-; and L 1 or L 2 The other two are -O(C=O)-, -(C=O)O-, -C(=O)-, -O-, and -S(O). x , -SS-, -C(=O)S-, SC(=O)-, -NR a C(=O)-、-C(=O)NR a -、NR a C(=O)NR a -、-OC(=O)NR a -or-NR a C(=O)O- or direct bond; G 1 and G 2 Each is an independent, unreplaced C1-C 12 Alkylene or C1-C 12 alkenyl; G 3 It is C1-C 24 Alkylene, C1-C 24 alkenyl, C3-C8 cycloalkylene, C3-C8 cycloalkenyl; R a Is it H or C1-C? 12 Alkyl; R 1 and R 2 Each is independently C6-C 24 Alkyl or C6-C 24 alkenyl; R 3 Is it H or OR? 5 CN, -C(=O)OR 4 -OC(=O)R 4 or -NR 5 C(=O)R 4 ;R 4 It is C1-C 12 Alkyl; R 5 It is H or C1-C6 alkyl; and x is 0, 1 or 2.

[0557] Other suitable additional cationic lipids for use in the compositions include cationic lipids as described in International Patent Publication WO2017 / 117528, which is incorporated herein by reference. In some embodiments, these compositions comprise cationic lipids having the following compound structures:

[0558]

[0559] And its pharmaceutically acceptable salts.

[0560] In some embodiments, these compositions comprise cationic lipids having the following compound structures:

[0561]

[0562] And its pharmaceutically acceptable salts.

[0563] In some embodiments, these compositions comprise cationic lipids having the following compound structures:

[0564]

[0565] And its pharmaceutically acceptable salts.

[0566] Other suitable additional cationic lipids for use in the composition include cationic lipids as described in International Patent Publication WO2017 / 049245, which is incorporated herein by reference. In some embodiments, the cationic lipids in the compositions and methods of the present invention comprise compounds having one of the following formulas:

[0567] ,

[0568] ,

[0569] ,as well as

[0570] ,

[0571] And its pharmaceutically acceptable salts. With respect to any of these four formulas, R4 is independently selected from -(CH2). n Q and -(CH2) n CHQR; Q can be selected freely from -OR, -OH, or -O(CH2). n The group consisting of N(R)2, -OC(O)R, -CX3, -CN, -N(R)C(O)R, -N(H)C(O)R, -N(R)S(O)2R, -N(H)S(O)2R, -N(R)C(O)N(R)2, -N(H)C(O)N(R)2, -N(H)C(O)N(H)(R), -N(R)C(S)N(R)2, -N(H)C(S)N(R)2, -N(H)C(S)N(H)(R) and heterocyclic rings; R is independently selected from the group consisting of: C 1-3 Alkyl, C 2-3 Alkenyl and H; and n is 1, 2 or 3.

[0572] In some embodiments, these compositions comprise cationic lipids having the following compound structures:

[0573]

[0574] And its pharmaceutically acceptable salts.

[0575] In some embodiments, these compositions comprise cationic lipids having the following compound structures:

[0576]

[0577] And its pharmaceutically acceptable salts.

[0578] In some embodiments, these compositions comprise cationic lipids having the following compound structures:

[0579]

[0580] And its pharmaceutically acceptable salts.

[0581] In some embodiments, these compositions comprise cationic lipids having the following compound structures:

[0582]

[0583] And its pharmaceutically acceptable salts.

[0584] Other suitable additional cationic lipids for use in the composition include cationic lipids as described in International Patent Publications WO2017 / 173054 and WO 2015 / 095340, which are incorporated herein by reference.

[0585] In some embodiments, these compositions comprise cationic lipids having the following compound structures:

[0586]

[0587] And its pharmaceutically acceptable salts.

[0588] In some embodiments, these compositions comprise cationic lipids having the following compound structures:

[0589]

[0590] And its pharmaceutically acceptable salts.

[0591] In some embodiments, these compositions comprise cationic lipids having the following compound structures:

[0592]

[0593] And its pharmaceutically acceptable salts.

[0594] In some embodiments, these compositions comprise cationic lipids having the following compound structures:

[0595]

[0596] And its pharmaceutically acceptable salts.

[0597] Other suitable additional cationic lipids for use in the composition include cholesterol-based cationic lipids. In some embodiments, these compositions comprise imidazole cholesterol esters or "ICE" having the following compound structure:

[0598] (ICE)

[0599] And its pharmaceutically acceptable salts.

[0600] Other suitable additional cationic lipids for use in the composition include cleavable cationic lipids as described in International Patent Publication WO2012 / 170889, which is incorporated herein by reference. In some embodiments, these compositions comprise cationic lipids having the following formula:

[0601] ,

[0602] R1 is selected from the group consisting of imidazole, guanidine, amino, imine, enamine, optionally substituted alkylamino (e.g., alkylamino, such as dimethylamino) and pyridyl; R2 is selected from the group consisting of one of the following two formulas:

[0603] and

[0604] Furthermore, R3 and R4 are each independently selected from the group consisting of: optionally substituted variable saturated or unsaturated C6-C. 20 Alkyl groups and optionally substituted C6-C groups that are variablely saturated or unsaturated 20 Acyl group; and where n is zero or any positive integer (e.g., one, two, three, four, five, six, seven, eight, nine, ten, eleven, twelve, thirteen, fourteen, fifteen, sixteen, seventeen, eighteen, nineteen, twenty or greater).

[0605] In some embodiments, these compositions comprise a cationic lipid “HGT4001” having the following compound structure:

[0606] (HGT4001)

[0607] And its pharmaceutically acceptable salts.

[0608] In some embodiments, these compositions comprise a cationic lipid “HGT4002” having the following compound structure:

[0609] (HGT4002)

[0610] And its pharmaceutically acceptable salts.

[0611] In some embodiments, these compositions comprise a cationic lipid “HGT4003” having the following compound structure:

[0612] (HGT4003)

[0613] And its pharmaceutically acceptable salts.

[0614] In some embodiments, these compositions comprise a cationic lipid “HGT4004” having the following compound structure:

[0615] (HGT4004)

[0616] And its pharmaceutically acceptable salts.

[0617] In some embodiments, these compositions comprise a cationic lipid “HGT4005” having the following compound structure:

[0618] (HGT4005)

[0619] And its pharmaceutically acceptable salts.

[0620] In some embodiments, these compositions comprise the cationic lipid N-[l-(2,3-dioleoxy)propyl]-N,N,N-trimethylammonium chloride (“DOTMA”). Feigner et al. (Proc. Nat'l Acad. Sci. [Proceedings of the National Academy of Sciences] 84, 7413 (1987); U.S. Patent No. 4,897,355, each of which is incorporated herein by reference. DOTMA can be formulated alone or in combination with neutral lipids (e.g., dioleoylphosphatidylethanolamine or "DOPE") or other cationic or noncationic lipids into liposome transfer mediators or lipid nanoparticles, and such liposomes can be used to enhance the delivery of nucleic acids to target cells. Other cationic lipids suitable for the composition include, for example, 5-carboxy-spermine-glycine bis(octadecylamide) ("DOGS"); 2,3-dioleoyloxy-N-[2(spermine-formylamino)ethyl]-N,N-dimethyl-1-propylaminoonium ("DOSPA") (Behr et al. Proc. Nat'l Acad. Sci. [Proceedings of the National Academy of Sciences] 86, 6982). (1989), U.S. Patent No. 5,171,678; U.S. Patent No. 5,334,761); 1,2-dioleoyl-3-dimethylammonium-propane (“DODAP”); 1,2-dioleoyl-3-trimethylammonium-propane (“DOTAP”).

[0621] Other exemplary cationic lipids suitable for the composition include: 1,2-distearyloxy-N,N-dimethyl-3-aminopropane (“DSDMA”); 1,2-dioleyloxy-N,N-dimethyl-3-aminopropane (“DODMA”); 1,2-dilinoleyloxy-N,N-dimethyl-3-aminopropane (“DLinDMA”); 1,2-di-linoleyloxy-N,N-dimethyl-3-aminopropane (“DLenDMA”); N-dioleyl-N,N-dimethylammonium chloride (“DODAC”); N,N -Distearyl-N,N-dimethylammonium bromide (“DDAB”); N-(1,2-dimyristoyloxypropyl-3-yl)-N,N-dimethyl-N-hydroxyethylammonium bromide (“DMRIE”); 3-dimethylamino-2-(cholest-5-en-3-β-oxybut-4-oxy)-1-(cis,cis-9,12-octadecadienoxy)propane (“CLinDMA”); 2-[5'-(cholest-5-en-3-β-oxy)-3'-oxaproxy)-3-dimethyl-l-(cis,cis-9',l-2'-octadecadienoxy)propane Carbodienoxy)propane (“CpLinDMA”); N,N-dimethyl-3,4-dioleoyloxybenzylamine (“DMOBA”); 1,2-N,N'-dioleoylcarbamoyl-3-dimethylaminopropane (“DOcarbDAP”); 2,3-di-linoleyloxy-N,N-dimethylpropylamine (“DLinDAP”); 1,2-N,N'-dilinoleoylcarbamoyl-3-dimethylaminopropane (“DLincarbDAP”); 1,2-dilinoleoylcarbamoyl-3-dimethylaminopropane (“DLin”) CDAP”); 2,2-Dilinoleyl-4-dimethylaminomethyl-[1,3]-dioxacyclopentane (“DLin-K-DMA”); 2-((8-[(3β)-cholesterol-5-en-3-yloxy]octyl)oxy)-N,N-dimethyl-3-[(9Z,12Z)-octadec-9,12-dien-1-yloxy]propane-1-amine (“Octyl-CLinDMA”); (2R)-2-((8-[(3β)-cholesterol-5-en-3-yloxy]octyl)oxy)-N,N-dimethyl-3-[(9Z,12Z)-octadec-9,12-dien-1-yloxy]propane-1-amine (“Octyl-CLinDMA”); (2R)-2-((8-[(3β)-cholesterol-5-en-3-yloxy]octyl)oxy)-N,N-dimethyl-3-[(9Z,12Z)-1-propane-1-amine) (12Z)-octadec-9,12-dien-1-yloxy]prop-1-amine (“Octyl-CLinDMA(2R)”); (2S)-2-((8-[(3β)-cholesterol-5-en-3-yloxy]octyl)oxy)-N,fsl-dimethyl-3-[(9Z, 12Z)-octadec-9,12-dien-1-yloxy]prop-1-amine (“Octyl-CLinDMA(2S)”);2,2-Dilinoleoyl-4-dimethylaminoethyl-[1,3]-dioxacyclopentane (“DLin-K-XTC2-DMA”); and 2-(2,2-di((9Z,12Z)-octadec-9,12-dien-1-yl)-1,3-dioxacyclopentan-4-yl)-N,N-dimethylethylamine (“DLin-KC2-DMA”) (see WO 2010 / 042877, which is incorporated herein by reference; Semil et al., Nature Biotech. [Nature Biotech] 28: 172-176 (2010)). (Heyes, J. et al., J Controlled Release 107: 276-287 (2005); Morrissey, DV. et al., Nat. Biotechnol. 23(8): 1003-1007 (2005); International Patent Publication WO 2005 / 121348). In some embodiments, one or more of the cationic lipids comprise at least one of an imidazole, a dialkylamino, or a guanidine moiety.

[0622] In some embodiments, one or more cationic lipids suitable for the composition include 2,2-dilinole-4-dimethylaminoethyl-[1,3]-dioxacyclopentane (“XTC”); (3aR,5s,6aS)-N,N-dimethyl-2,2-bis((9Z,12Z)-octadec-9,12-dienyl)tetrahydro-3aH-cyclopenta[d][1,3]dioxacyclopenta-5-amine (“ALNY-100”); and / or 4,7,13-tris(3-oxo-3-(undecapylamino)propyl)-N1,N16-teicocarbato-4,7,10,13-tetraazahexadecane-1,16-diamide (“NC98-5”).

[0623] In some embodiments, the percentage of total cationic lipids in the composition (e.g., a liposome composition) may be no more than 10%, no more than 20%, no more than 30%, no more than 40%, no more than 50%, no more than 60%, no more than 70%, no more than 80%, no more than 90%, or no more than 95% of the total lipids, as measured by molar ratio (mol%) or by weight (wt%).

[0624] In some embodiments, the percentage of total cationic lipids in the composition (e.g., a liposome composition) may be greater than 10%, greater than 20%, greater than 30%, greater than 40%, greater than 50%, greater than 60%, greater than 70%, greater than 80%, greater than 90%, or greater than 95% of the total lipids, as measured by molar ratio (mol%) or by weight (wt%).

[0625] In some embodiments, by weight, total cationic lipids constitute about 30%-50% of the liposomes (e.g., about 30%-45%, about 30%-40%, about 35%-50%, about 35%-45%, or about 35%-40%). In some embodiments, by molar ratio, cationic lipids constitute about 30%, about 35%, about 40%, about 45%, or about 50% of the composition (e.g., the liposome composition). In some embodiments, by weight, total cationic lipids constitute about 30%-50% of the liposomes (e.g., about 30%-45%, about 30%-40%, about 35%-50%, about 35%-45%, or about 35%-40%). In some embodiments, by weight, cationic lipids constitute about 30%, about 35%, about 40%, about 45%, or about 50% of the composition (e.g., the liposome composition).

[0626] The composition (e.g., an LPN composition) may also contain one or more non-cationic (“auxiliary”) lipids. As used herein, the phrase “non-cationic lipid” refers to any neutral, zwitterionic, or anionic lipid. As used herein, the phrase “anionic lipid” refers to any of a variety of lipid substances that carry a net negative charge at a selected pH (such as physiological pH). Non-cationic lipids include, but are not limited to, distearylphosphatidylcholine (DSPC), dioleoylphosphatidylcholine (DOPC), dipalmitoylphosphatidylcholine (DPPC), dioleoylphosphatidylglycerol (DOPG), dipalmitoylphosphatidylglycerol (DPPG), dioleoylphosphatidylethanolamine (DOPE), palmitoyloleoylphosphatidylcholine (POPC), palmitoyloleoyl-phosphatidylethanolamine (POPE), dioleoyl-phosphatidylethanolamine 4-(N-maleimidemethyl)-cyclohexane-l-carboxylate (DOPE-mal), dipalmitoylphosphatidylethanolamine (DPPE), dimyristoylphosphatidylethanolamine (DMPE), distearyl-phosphatidyl-ethanolamine (DSPE), 16-O-monomethylPE, 16-O-dimethylPE, 18-1-transPE, l-stearoyl-2-oleoyl-phosphatidylethanolamine (SOPE), or mixtures thereof.

[0627] In the examples, the non-cationic lipid or auxiliary lipid is dioleoylphosphatidylethanolamine (DOPE).

[0628] 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.

[0629] In some embodiments, non-cationic lipids may be present in the following molar ratios (mol%): 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, total non-cationic lipids may be present in the following molar ratios (mol%): 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%. 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 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 liposomes may be no more than about 5 mol%, no more than about 10 mol%, no more than about 20 mol%, no more than about 30 mol%, or no more than about 40 mol%. In some embodiments, the percentage of total non-cationic lipids in liposomes may be no more than about 5 mol%, no more than about 10 mol%, no more than about 20 mol%, no more than about 30 mol%, or no more than about 40 mol%.

[0630] In some embodiments, non-cationic lipids may be present in the following weight percentages (wt%): 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, total non-cationic lipids may be present in the following weight percentages (wt%): 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%. In some embodiments, the percentage of non-cationic lipids in the liposomes 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 total non-cationic lipids in liposomes may be greater than about 5 wt%, greater than about 10 wt%, greater than about 20 wt%, greater than about 30 wt%, or greater than about 40 wt%. In some embodiments, the percentage of non-cationic lipids in liposomes may be no more than about 5 wt%, no more than about 10 wt%, no more than about 20 wt%, no more than about 30 wt%, or no more than about 40 wt%. In some embodiments, the percentage of total non-cationic lipids in liposomes may be no more than about 5 wt%, no more than about 10 wt%, no more than about 20 wt%, no more than about 30 wt%, or no more than about 40 wt%.

[0631] Structural lipids

[0632] The structural lipid component provides stability to the lipid bilayer structure within the lipid nanoparticles. In some embodiments, the LNP comprises one or more structural lipids. In some embodiments, the structural lipids are cholesterol-based lipids. Suitable cholesterol-based lipids include, for example: DC-Choi (N,N-dimethyl-N-ethylformamidocholesterol), 1,4-bis(3-N-oleenylaminopropyl)piperazine (Gao et al., Biochem Biophys Res Comm. [Biochemical and Biophysical Research Communications] (1991) 179:280; Wolf et al., BioTechniques [Biotechnology] (1997) 23:139; US Patent 5,744,335), imidazole cholesterol ester (“ICE”; WO 2011 / 068810), sitosterol (22,23-dihydrostigmasterol), β-stigmasterol, sitosterol, fucosterol, stigmasterol (stigmasterane-5,22-dien-3-ol), ergosterol, lanosterol (3β-hydroxy-5,24-cholestadiene), lanosterol (8,24-lanosterdien-3β-ol), 7-dehydrocholesterol (Δ5,7-cholesterol), dihydrolanosterol (24,25-dihydrolanosterol), yeast sterol (5α-cholest-8,24-dien-3β-ol) ), cholesterol (5α-cholest-7-en-3β-ol), diosgenin ((3β,25R)-spirost-5-en-3-ol), campesterol (campester-5-en-3β-ol), campesteranol (5a-campesteran-3b-ol), 24-methylene cholesterol (5,24(28)-cholestadien-24-methylene-3β-ol), heptadecanol (heptadecanol-5-en-3β-yl ester), cholesterol oleate, cholesterol stearate and other modified cholesterol forms. In some embodiments, suitable cholesterol-based lipids include cholesterol and, for example, DC-Chol (N,N-dimethyl-N-ethylformamidocholesterol), 1,4-bis(3-N-oleoylaminopropyl)piperazine (Gao et al. Biochem. Biophys. Res. Comm. [Biochemistry and Biophysics Research Communications] 179, 280 (1991); Wolf et al. BioTechniques [Biotechnology] 23, 139 (1997); U.S. Patent No. 5,744,335), or imidazole cholesterol ester (ICE) having the following structure.

[0633] ("ICE").

[0634] In the embodiments, the cholesterol-based lipid is cholesterol.

[0635] In some embodiments, cholesterol-based lipids may be present in the following molar ratios (mol%): about 1% to about 30% of the total lipids present in the liposomes, or about 5% to about 20%. 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 be no more than about 5 mol%, no more than about 10 mol%, no more than about 20 mol%, no more than about 30 mol%, or no more than about 40 mol%.

[0636] In some embodiments, cholesterol-based lipids may be present in the following weight percentages (wt%): about 1% to about 30% of the total lipids present in the liposomes, or about 5% to about 20%. 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 be no more than about 5 wt%, no more than about 10 wt%, no more than about 20 wt%, no more than about 30 wt%, or no more than about 40 wt%.

[0637] Hidden lipids

[0638] Stealth lipid components provide control over the particle size and stability of nanoparticles. The addition of such components can prevent complex aggregation and provides a means to increase the cycle life of lipid-nucleic acid drug compositions and enhance their delivery to target tissues.

[0639] In some embodiments, the stealth lipids are polyethylene glycol-conjugated (PEGylated) lipids. These components can be selected to rapidly exchange the pharmaceutical composition in vivo (see, for example, U.S. Patent 5,885,613).

[0640] The PEGylated lipids considered include, but are not limited to, polyethylene glycol (PEG) chains up to 5 kDa in length, covalently attached to one or more alkyl chains having a length of C6-C20 (e.g., C8, C10, C12, C14, C16, or C18), such as derivatized ceramides (e.g., N-octanoyl-sphingosine-1-[succinyl(methoxy)polyethylene glycol)] (C8 PEG ceramide)). In some embodiments, the PEGylated lipids are 1,2-dimyristoyl-racemic-glycerol-3-methoxypolyethylene glycol (DMG-PEG); 1,2-distearyl-sn-glycerol-3-phosphate ethanolamine-polyethylene glycol (DSPE-PEG); 1,2-dilauryl-sn-glycerol-3-phosphate ethanolamine-polyethylene glycol (DLPE-PEG); or 1,2-distearyl-racemic-glycerol-polyethylene glycol (DSG-PEG).

[0641] In some embodiments, the PEG has a high molecular weight, for example, 2000-2400 g / mol. In some embodiments, the PEG is PEG2000 (or PEG-2K). In some embodiments, the PEGylated lipid is DMG-PEG2000, DSPE-PEG2000, DLPE-PEG2000, DSG-PEG2000, or C8 PEG2000. In some embodiments, the PEGylated lipid is dimyristoyl-PEG2000 (DMG-PEG2000).

[0642] In some embodiments, the stealth lipid is a polyoxazoline polymer-conjugated lipid. Polyoxazoline polymer-conjugated lipids suitable for the LNP compositions disclosed herein are described, for example, in WO 2022 / 173667 and WO 2023 / 031394.

[0643] In some embodiments, the stealth lipid is a polysarcosine-conjugated (pSar) lipid. In some embodiments, the polysarcosine comprises 25-45 sarcosine units. In some embodiments, the polysarcosine comprises 25 sarcosine units. In some embodiments, the polysarcosine comprises 35 sarcosine units. In some embodiments, the polysarcosine comprises 45 sarcosine units. Non-limiting examples of pSar lipids include N-tetradecyl-pSar25, N-hexadecyl-pSar25, N-octadecyl-pSar25, N-dodecyl-pSar25, 1,2-dimyristoyl-sn-glycerol-3-succinoyl-N-polysarcosine-25 (DMG-pSar25), 1,2-dioleoyl-sn-glycerol-3-phosphoethanolamine-N-polysarcosine-25 (18:1 PE (DOPE) pSar25), N,N-tetracosylamine-N-succinoyl[methyl(polysarcosine)45], N,N-tetracosylamine-N-succinoyl[methyl(polysarcosine)35], and N,N-tetracosyl-polysarcosine-25. Further examples of pSar lipids suitable for the LNP compositions disclosed herein are described in WO 2020 / 070040.

[0644] In some embodiments, the composition (e.g., an LPN composition) comprises one or more PEGylated lipids.

[0645] For example, the present invention also envisions the use of polyethylene glycol (PEG) modified phospholipids and derived lipids (such as derived ceramides (PEG-CER), including N-octanoyl-sphingosine-1-[succinyl(methoxypolyethylene glycol)-2000] (C8 PEG-2000 ceramide)) in combination with one or more cationic lipids, and in some embodiments, in combination with other lipids comprising liposomes. In some embodiments, particularly usable exchangeable lipids are those having shorter acyl chains (e.g., C8 PEG-2000 ceramide). 14 Or C 18 PEG-ceramide.

[0646] In the examples, the PEG-modified lipid is 1,2-dimyristoyl-sn-glycerol, methoxy polyethylene glycol (DMG-PEG2000).

[0647] The envisioned PEG-modified lipids (also referred to herein as PEGylated lipids, a term that may be used interchangeably with PEG-modified lipids) include, but are not limited to, those with C6-C... 20A polyethylene glycol chain up to 5 kDa covalently attached to one or more alkyl chains of a length. In some embodiments, the PEG-modified or PEGylated lipid is 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 enhancing the delivery of lipid-nucleic acid compositions to target cells (Klibanov et al. (1990) FEBS Letters [European Federation of Biochemical Societies Letters], 268 (1): 235-237), or they can be selected for rapid in vivo exchange of formulations (see U.S. Patent No. 5,885,613).

[0648] The PEG-modified phospholipids and derived lipids of the present invention may be present in a molar ratio (mol%) of about 0% to about 15%, about 0.5% to about 15%, about 1% to about 15%, about 4% to about 10%, or about 2% of the total lipids present in the composition (e.g., liposome composition).

[0649] The PEG-modified phospholipids and derived lipids of the present invention may be present in weight percentages (wt%) of about 0% to about 15%, about 0.5% to about 15%, about 1% to about 15%, about 4% to about 10%, or about 2% of the total lipids present in the composition (e.g., liposome composition).

[0650] Pharmaceutical preparations and therapeutic uses

[0651] According to one purpose, the present invention relates to a pharmaceutical composition comprising an LNP according to the invention, and one or more pharmaceutically acceptable excipients.

[0652] According to another objective, the present invention also relates to an LNP according to the invention for delivering nucleic acids into a patient's cells.

[0653] In an embodiment, the LNP can be used as a vaccine, such as a vaccine against influenza or a respiratory syncytial virus (RSV) vaccine.

[0654] Lipids having formula (I) as described herein can be used to prepare compositions (e.g., LPN compositions) that promote or enhance the delivery and release of encapsulated materials (e.g., one or more therapeutic polynucleotides) to one or more target cells (e.g., by permeation or fusion with the lipid membrane of such target cells).

[0655] For example, when an LPN composition (e.g., liposome nanoparticles) contains or otherwise enriches one or more of the compounds of formula (I) disclosed herein, a phase transition in the lipid bilayer of the one or more target cells can facilitate the delivery of encapsulated material (e.g., one or more therapeutic polynucleotides encapsulated in lipid nanoparticles) into the one or more target cells.

[0656] Similarly, in some embodiments, lipids having formula (I) as described herein can be used to prepare liposomal mediators 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, allowing for the administration of reduced amounts of such compositions to subjects to achieve the desired therapeutic response or outcome.

[0657] Therefore, pharmaceutical formulations comprising lipids of formula (I) as described herein and nucleic acids provided by the present invention can be used for a variety of therapeutic purposes. To facilitate the delivery of nucleic acids in vivo, lipids of formula (I) as described herein and nucleic acids can be formulated in combination with one or more additional drug carriers, targeting ligands, or stabilizing agents. In some embodiments, lipids of formula (I) as described herein can be formulated via a premixed lipid solution. In other embodiments, a post-insertion technique can be used to formulate compositions comprising lipids of formula (I) as described herein into lipid membranes of nanoparticles. Pharmaceutical formulation and administration techniques can be found in the latest edition of “Remington's Pharmaceutical Sciences”, Mack Publishing Co., Easton, Pa.

[0658] Suitable routes of administration include, for example, oral, rectal, vaginal, transmucosal, pulmonary (including tracheal or inhalation), or enteral administration; parenteral delivery, including intradermal, percutaneous (local), intramuscular, subcutaneous, intramedullary injection, as well as intrathecal, direct intravenous, intravenous, intraperitoneal, or intranasal administration.

[0659] In the embodiments, the composition can be formulated for intravenous (IV) administration.

[0660] In the embodiments, the composition can be formulated for intramuscular (IM) administration.

[0661] In the embodiments, the composition can be formulated for oral administration.

[0662] In the embodiments, the composition may be formulated for administration by inhalation (e.g., the composition may be formulated for nebulization).

[0663] In certain embodiments, intramuscular administration is directed against a muscle selected from the group consisting of skeletal muscle, smooth muscle, and cardiac muscle. In some embodiments, administration results in the delivery of nucleic acids to muscle cells. In some embodiments, administration results in the delivery of nucleic acids to hepatocytes (i.e., liver cells). In some embodiments, administration is intramuscular. In some embodiments, administration is intravenous.

[0664] Alternatively or additionally, the pharmaceutical formulations of the present invention can be administered locally rather than systemically, for example, by direct injection of the pharmaceutical formulation into the targeted tissue (preferably in a sustained-release formulation). Depending on the tissue to be targeted, local delivery can be achieved in various ways. Exemplary tissues in which the delivered and / or expressed mRNA can be delivered include, but are not limited to, the liver, kidneys, heart, spleen, serum, brain, skeletal muscle, lymph nodes, skin, and / or cerebrospinal fluid. In an example, the targeted 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); for example, 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; can be provided in the form of liquids, tablets, or capsules for gastric or intestinal administration; can be provided in the form of suppositories for rectal or vaginal application; or can even be delivered to the eyes using creams, drops, or even injections.

[0665] In embodiments, administration is via pulmonary delivery. As used herein, pulmonary delivery refers to delivery to the lungs via, for example, the nasal cavity, trachea, bronchi, bronchioles, and / or other pulmonary systems. In embodiments, the compositions described herein are formulated for nebulization. In embodiments, the delivery medium may be in a nebulized composition that can be inhaled. In embodiments, pulmonary delivery involves inhalation (e.g., for nasal, tracheal, or bronchial delivery). In embodiments, the composition is nebulized prior to inhalation.

[0666] This invention provides a method for delivering a composition having a full-length mRNA molecule encoding a target peptide or polypeptide for treating a subject, such as a human subject or cells of a human subject, or cells of a human subject that are being treated and delivered to the human subject's cells. In some aspects, the invention is characterized by methods for preventing and / or treating a disease in a subject, methods comprising administering to the subject the composition as described herein (e.g., a pharmaceutical composition).

[0667] The compositions described herein may contain mRNA encoding peptides, including those described herein (e.g., polypeptides such as proteins).

[0668] In the examples, mRNA encodes a polypeptide.

[0669] In this embodiment, the mRNA encodes a peptide. In this embodiment, the peptide is an antigen.

[0670] In the examples, mRNA encodes proteins.

[0671] The present invention provides a method for delivering a composition having a full-length mRNA molecule encoding a target peptide or protein for treating a subject, such as a human subject or a cell of a human subject, or for treating and delivering to a cell of a human subject.

[0672] In some embodiments, the present invention provides a method for producing a therapeutic composition having full-length mRNA encoding a peptide or polypeptide for delivering a vaccine to a subject or the subject's cells or for treating the subject with a vaccine. For example, in some embodiments, the present invention provides a method for producing a therapeutic composition having full-length mRNA encoding an antigen derived from an infectious agent, such as a virus. In some embodiments, the present invention provides a method for producing a therapeutic composition having full-length mRNA encoding an antigen derived from an influenza virus or RSV.

[0673] Nucleic acid synthesis

[0674] The nucleic acids according to the invention can be synthesized according to any known method. For example, the mRNA according to the invention can be synthesized via in vitro transcription (IVT). In short, IVT is typically performed using: a linear or circular DNA template containing a promoter, a ribonucleotide triphosphate pool, a buffer system that may include 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.

[0675] 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 (e.g., T3, T7, mutated T7, or SP6 promoter), followed by the desired nucleotide sequence for the desired mRNA and a termination signal.

[0676] One or more desired mRNA sequences according to the invention can be determined and incorporated into a DNA template using standard methods. For example, starting with the desired amino acid sequence (e.g., an enzyme sequence), a virtual reverse translation is performed based on the degenerate genetic code. An optimization algorithm can then be used to select suitable codons. Typically, in one aspect, the G / C content can be optimized to achieve the highest possible G / C content, and in another aspect, the frequency of tRNA can be optimally considered based on codon usage. The optimized RNA sequence can be established and displayed, for example, by means of a suitable display device, and compared with the original (wild-type) sequence. Secondary structures can also be analyzed to calculate stable and unstable properties, or the regions of RNA can be calculated separately.

[0677] As stated above, the term "nucleic acid" in its broadest sense refers to any compound and / or substance incorporated into or potentially incorporated into a polynucleotide chain. DNA can be in the form of antisense DNA, plasmid DNA, portions of plasmid DNA, pre-condensed DNA, polymerase chain reaction (PCR) products, vectors (e.g., P1, PAC, BAC, YAC, artificial chromosomes), expression cassettes, chimeric sequences, chromosomal DNA, or derivatives of these groups. RNA can be in the form of messenger RNA (mRNA), ribosomal RNA (rRNA), signal recognition particle RNA (7SL RNA or SRP RNA), transfer RNA (tRNA), transfer messenger RNA (tmRNA), small nuclear RNA (snRNA), small nucleolar RNA (snoRNA), SmY RNA, small Cahalosome-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 non-coding RNA (lncRNA), microRNA (miRNA), piwi-interacting RNA (piRNA), small interfering RNA (siRNA), transcribed siRNA (tasiRNA), repeat-associated siRNA (rasiRNA), 73K RNA, retrotransposons, viral genomes, viroids, satellite RNA, or derivatives of these groups. In some embodiments, the nucleic acid is mRNA encoding a peptide, polypeptide, or protein.

[0678] In some embodiments, mRNA encoding peptides or polypeptides is used in vaccines. In some embodiments, the mRNA encodes an antigen (e.g., an antigen derived from an infectious agent).

[0679] mRNA synthesis

[0680] The mRNA according to the invention can be synthesized according to any of a variety of known methods. For example, the mRNA according to the invention can be synthesized via in vitro transcription (IVT). Briefly, IVT is typically performed using: a linear or circular DNA template containing a promoter, a ribonucleotide triphosphate pool, a buffer system including DTT and magnesium ions, and a suitable RNA polymerase (e.g., T3, T7, or SP6 RNA polymerase), DNase I, pyrophosphatase, and / or an RNase inhibitor. The exact conditions will vary depending on the specific application. The presence of these reagents is undesirable in the final product according to several embodiments and can therefore be referred to as an impurity, and preparations containing one or more of these impurities can be referred to as impure preparations. In some embodiments, in vitro transcription occurs in single batches.

[0681] 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 (e.g., a T3, T7, or SP6 promoter), followed by the desired nucleotide sequence for the desired mRNA and a termination signal.

[0682] One or more desired mRNA sequences according to the invention can be determined and incorporated into a DNA template using standard methods. For example, starting with the desired amino acid sequence (e.g., an enzyme sequence), a virtual reverse translation is performed based on the degenerate genetic code. An optimization algorithm can then be used to select suitable codons. Typically, in one aspect, the G / C content can be optimized to achieve the highest possible G / C content, and in another aspect, the frequency of tRNA can be optimally considered based on codon usage. The optimized RNA sequence can be established and displayed, for example, by means of a suitable display device, and compared with the original (wild-type) sequence. Secondary structures can also be analyzed to calculate stable and unstable properties, or the regions of RNA can be calculated separately.

[0683] Modified mRNA

[0684] In some embodiments, the mRNA according to the invention can be synthesized as unmodified or modified mRNA. Modified mRNA includes nucleotide modifications in 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, mRNA 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)) and modified nucleotides, purine and pyrimidine analogs or derivatives, 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-thio-cytosine, etc. 4-Thiouracil, 5-Carboxymethylaminomethyl-2-thiouracil, 5-(Carboxyhydroxymethyl)-uracil, 5-Fluorouracil, 5-Bromouracil, 5-Carboxymethylaminomethyl-uracil, 5-Methyl-2-thiouracil, 5-Methyluracil, N-uracil-5-oxyacetic acid methyl ester, 5-Methylaminomethyl-uracil, 5-Methoxyaminomethyl-2-thiouracil Uracil, 5'-methoxycarbonylmethyl-uracil, 5-methoxy-uracil, uracil-5-oxyacetic acid methyl ester, uracil-5-oxyacetic acid (v), 1-methyl-pseudouracil, queosine, β-D-mannosyl-queosine, wybutoxosine, and phosphoramide, thiophosphate, peptide nucleotide, methylphosphonate, 7-denitroguanosine, 5-methylcytosine, and inosine. Those skilled in the art will know the preparation of such analogues, for example, from the following documents: 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.

[0685] In some embodiments, mRNA may contain RNA backbone modifications. Typically, backbone modifications are modifications in which the phosphate esters of the nucleotide backbone contained in the RNA are chemically modified. Exemplary backbone modifications typically include, but are not limited to, modifications from the group consisting of: methylphosphonates, methylphosphonamides, phosphoramides, thiophosphates (e.g., cytidine 5'-O-(1-thiophosphate)), borate phosphates, positively charged guanidinium groups, etc., which means by replacing the phosphodiester bonds with other anionic, cationic, or neutral groups.

[0686] In some embodiments, mRNA may contain sugar modifications. Typical sugar modifications are chemical modifications of the sugars in the nucleotides it contains, including but not limited to sugar modifications selected from the group consisting of: 4'-thio-ribonucleotides (see, for example, U.S. Patent Application Publication No. US 2016 / 0031928, which is incorporated herein by reference), 2'-deoxy-2'-fluoro-oligonucleotides (2'-fluoro-2'-deoxycytidine 5'-triphosphate, 2'-fluoro-2'-deoxyuridine 5'-triphosphate), 2'-deoxy-2'-deamin-oligonucleotides (2'-amino-2'-deoxycytidine 5'-triphosphate, 2'-amino-2'-deoxyuridine 5'-triphosphate), 2'-O-alkyl oligonucleotides, 2'- Deoxy-2'-C-alkyl oligonucleotides (2'-O-methylcytidine 5'-triphosphate, 2'-methyluridine 5'-triphosphate), 2'-C-alkyl oligonucleotides, and their isomers (2'-aracytidine 5'-triphosphate, 2'-arylurogidine 5'-triphosphate), or azidotriphosphates (2'-azido-2'-deoxycytidine 5'-triphosphate, 2'-azido-2'-deoxyuridine 5'-triphosphate).

[0687] In some embodiments, mRNA may contain modifications of nucleotide bases (base modifications). Modified nucleotides containing base modifications are also referred to as base-modified nucleotides. Examples of such base-modified nucleotides include, but are not limited to, 2-amino-6-chloropurine nucleotide 5'-triphosphate, 2-aminoadenosine 5'-triphosphate, 2-thiocytidine 5'-triphosphate, 2-thiouridine 5'-triphosphate, 4-thiouridine 5'-triphosphate, 5-aminoallylcytidine 5'-triphosphate, 5-aminoallyluridine 5'-triphosphate, 5-bromocytidine 5'-triphosphate, 5-bromouridine 5'-triphosphate, 5-iodocytidine 5'-triphosphate, 5-iodouridine 5'-triphosphate, 5-methylcytidine 5'-triphosphate, and 5-methyluridine 5'-triphosphate. Phosphate, 6-azacytidine 5'-triphosphate, 6-azauridine 5'-triphosphate, 6-chloropurine nucleoside 5'-triphosphate, 7-deadenosine 5'-triphosphate, 7-deadenosine 5'-triphosphate, 8-azaadenosine 5'-triphosphate, 8-azidoadenosine 5'-triphosphate, benzimidazole nucleoside 5'-triphosphate, N1-methyladenosine 5'-triphosphate, N1-methylguanosine 5'-triphosphate, N6-methyladenosine 5'-triphosphate, O6-methylguanosine 5'-triphosphate, pseudouridine 5'-triphosphate, puromycin 5'-triphosphate, or flavin 5'-triphosphate.

[0688] Typically, mRNA synthesis involves adding a "cap" at the N-terminus (5') and a "tail" at the C-terminus (3'). The presence of the cap is important in providing resistance to nucleases found in most eukaryotic cells. The presence of the tail protects the mRNA from degradation by exonucleases.

[0689] Therefore, in some embodiments, the mRNA includes a 5' cap structure. Typically, the 5' cap is added as follows: first, an RNA terminal phosphatase removes one terminal phosphate group from the 5' nucleotide, leaving two terminal phosphate groups; then, guanosine triphosphate (GTP) is added to the terminal phosphate group via guanylate transferase, creating a 5'5'5 triphosphate bond; then, the 7-nitrogen of guanine is methylated by a methyltransferase. Examples of cap structures include, but are not limited to, m7G(5')ppp(5'(A,G(5')ppp(5')A and G(5')ppp(5')G.

[0690] In some embodiments, the mRNA includes a 3' poly(A) tail. The poly(A) tail at the 3' end of the mRNA typically includes about 10 to 300 adenosine nucleotides (e.g., about 10 to 200 adenosine nucleotides, about 10 to 150 adenosine nucleotides, about 10 to 100 adenosine nucleotides, about 20 to 70 adenosine nucleotides, or about 20 to 60 adenosine nucleotides). In some embodiments, the mRNA includes a 3' poly(C) tail. A suitable poly(C) tail at the 3' end of the mRNA typically includes about 10 to 200 cytosine nucleotides (e.g., about 10 to 150 cytosine nucleotides, about 10 to 100 cytosine nucleotides, about 20 to 70 cytosine nucleotides, about 20 to 60 cytosine nucleotides, or about 10 to 40 cytosine nucleotides). The poly(C) tail may be added to the poly(A) tail or may replace the poly(A) tail.

[0691] In some embodiments, the mRNA includes a 5' and / or 3' untranslated region. In some embodiments, the 5' untranslated region includes one or more elements that affect the stability or translation of the mRNA, such as iron response elements. In some embodiments, the length of the 5' untranslated region may be between about 50 and 500 nucleotides.

[0692] In some embodiments, the 3' untranslated region includes one or more of the following: a polyadenylation signal, a protein binding site that affects the positional stability of mRNA in the cell, or one or more binding sites of miRNA. In some embodiments, the length of the 3' untranslated region may be between 50 and 500 nucleotides or longer.

[0693] hat structure

[0694] In some embodiments, the mRNA includes a 5' cap structure. Typically, the 5' cap is added as follows: first, an RNA terminal phosphatase removes one terminal phosphate group from the 5' nucleotide, leaving two terminal phosphate groups; then, guanosine triphosphate (GTP) is added to the terminal phosphate group via guanylate transferase, creating a 5'5'5 triphosphate bond; then, the 7-nitrogen of guanine is methylated by a methyltransferase. Examples of cap structures include, but are not limited to, m7G(5')ppp(5'(A,G(5')ppp(5')A and G(5')ppp(5')G.

[0695] The naturally occurring cap structure contains 7-methylguanosine, which is linked to the 5' end of the first transcribed nucleotide via a triphosphate bridge, producing m 7The G(5')ppp(5')N dinucleotide cap, where N is any nucleoside. In vivo, the cap is enzymatically added. The cap is added to the cell nucleus and catalyzed by guanylate transferase. The cap is added to the 5' end of the RNA immediately after transcription initiation. The terminal nucleoside is typically guanosine and is in the opposite orientation to all other nucleotides, i.e., G(5')ppp(5')GpNpNp.

[0696] The common cap for mRNA produced through in vitro transcription is m 7 G(5')ppp(5')G, which has been used in vitro transcribed with dinucleotide caps by T7 or SP6 RNA polymerase to obtain RNA with a capped structure at its 5' end. Popular methods for the in vitro synthesis of capped mRNA employ m 7 Pre-formed dinucleotides in the form of G(5')ppp(5')G (“m 7 GpppG was used as the transcription initiator.

[0697] To date, the common form of synthetic dinucleotide caps used in in vitro translation experiments is the anti-reverse cap analog (“ARCA”) or a modified ARCA, which is typically a modified cap analog in which the 2' or 3' OH group is replaced by -OCH3.

[0698] Other cap analogues include, but are not limited to, chemical structures selected from the group consisting of: m 7 GpppG, m 7 GpppA, m 7 GpppC; unmethylated cap analogues (e.g., GpppG); dimethylated cap analogues (e.g., m 2, 7 GpppG), trimethylated cap analogues (e.g., m 2,2,7 GpppG), dimethylated symmetric cap analogues (e.g., m 7 Gpppm 7 G) or anti-reverse cap analogues (e.g., ARCA; m 7 , 2'Ome GpppG, m 72'd GpppG, m 7,3'Ome GpppG, m 7,3' dGpppG and its tetraphosphate derivatives (see, for example, Jemielity, J. et al., “Novel 'anti-reverse' cap analogs with superior translational properties”, RNA, 9: 1108-1122 (2003)).

[0699] In some embodiments, a suitable cap is 7-methylguanylic acid (“m 7 G”, which is linked to the 5'-end of the first transcribed nucleotide via a triphosphate bridge, produces m 7 G(5')ppp(5')N, where N is any nucleoside. The m used in the embodiments of the present invention... 7 The preferred embodiment of the G-cap is m 7 G(5')ppp(5')G.

[0700] In some embodiments, the cap is a cap 0 structure. The cap 0 structure lacks the 2'-O-methyl residues attached to the ribose at bases 1 and 2. In some embodiments, the cap is a cap 1 structure. The cap 1 structure has a 2'-O-methyl residue at base 2. In some embodiments, the cap is a cap 2 structure. The cap 2 structure has 2'-O-methyl residues attached to both bases 2 and 3.

[0701] Multiple m 7 G-cap analogues are known in the art, and many are commercially available. These include the m described above. 7 GpppG, and ARCA 3'-OCH3 and 2'-OCH3 cap analogs (Jemielity, J. et al., RNA, 9: 1108-1122 (2003)). Other cap analogs used in embodiments of the invention include N7-benzylated dinucleotide tetraphosphate analogs (described in Grudzien, E. et al., RNA, 10: 1479-1487 (2004)), thiophosphate cap analogs (described in Grudzien-Nogalska, E. et al., RNA, 13: 1745-1755 (2007)), and cap analogs (including biotinylated cap analogs) described in U.S. Patent Nos. 8,093,367 and 8,304,529, which are incorporated herein by reference.

[0702] Tail structure

[0703] Typically, the presence of a "tail" protects mRNA from exonuclease degradation. Poly-A tails are thought to stabilize both natural messenger RNA and synthetic sense RNA. Therefore, in some embodiments, long poly-A tails can be added to mRNA molecules to enhance RNA stability. A variety of techniques recognized in the art can be used to add poly-A tails. For example, long poly-A tails can be added to synthetic or in vitro transcribed RNA using poly-A polymerase (Yokoe et al., Nature Biotechnology. 1996; 14: 1252-1256). Transcription vectors can also encode long poly-A tails. Alternatively, poly-A tails can be added by transcription directly from PCR products. Poly-A tails can also be ligated to the 3' end of sense RNA using RNA ligase (see, for example, Molecular Cloning A Laboratory Manual, 2nd edition, edited by Sambrook, Fritsch, and Maniatis (Cold Spring Harbor Laboratory Press: 1991)).

[0704] In some embodiments, the mRNA includes a 3' poly(A) tail. Typically, the length of the poly(A) tail can be at least about 10, 50, 100, 200, 300, 400, or at least 500 nucleotides. In some embodiments, the 3' end of the mRNA typically includes about 10 to 300 adenosine nucleotides (e.g., about 10 to 200 adenosine nucleotides, about 10 to 150 adenosine nucleotides, about 10 to 100 adenosine nucleotides, about 20 to 70 adenosine nucleotides, or about 20 to 60 adenosine nucleotides). In some embodiments, the mRNA includes a 3' poly(C) tail. A suitable poly-C tail at the 3' end of mRNA typically comprises about 10 to 200 cytosine nucleotides (e.g., about 10 to 150 cytosine nucleotides, about 10 to 100 cytosine nucleotides, about 20 to 70 cytosine nucleotides, about 20 to 60 cytosine nucleotides, or about 10 to 40 cytosine nucleotides). The poly-C tail may be added to or may replace the poly-A tail.

[0705] In some embodiments, the length of the poly-A tail or poly-C tail is adjusted to control the stability of the modified sense mRNA molecule of the present invention, and thus control protein transcription. For example, since the length of the poly-A tail can affect the half-life of the sense mRNA molecule, the length of the poly-A tail can be adjusted to modify the level of mRNA resistance to nucleases, thereby controlling the timing of polynucleotide expression and / or peptide production in target cells.

[0706] 5' and 3' non-translation areas

[0707] In some embodiments, the mRNA includes a 5' and / or 3' untranslated region. In some embodiments, the 5' untranslated region includes one or more elements that affect the stability or translation of the mRNA, such as iron response elements. In some embodiments, the length of the 5' untranslated region may be between about 50 and 500 nucleotides.

[0708] In some embodiments, the 3' untranslated region includes one or more of the following: a polyadenylation signal, a protein binding site that affects the positional stability of mRNA in the cell, or one or more binding sites of miRNA. In some embodiments, the length of the 3' untranslated region may be between 50 and 500 nucleotides or longer.

[0709] Exemplary 3' and / or 5' UTR sequences can be derived from stable mRNA molecules (e.g., globin, actin, GAPDH, tubulin, histone, or citrate cycling enzymes) to increase the stability of sense mRNA molecules. For example, a 5' UTR sequence may include a portion of the CMV, i.e., the early 1 (IE1) gene, or a fragment thereof, to improve the nuclease resistance and / or half-life of the polynucleotide. Including a sequence encoding human growth hormone (hGH) or a fragment thereof at the 3' end or untranslated region of a polynucleotide (e.g., mRNA) is also considered to further stabilize the polynucleotide. Typically, these modifications improve the stability and / or pharmacokinetic properties (e.g., half-life) of the polynucleotide relative to its unmodified counterpart, and include modifications, for example, to improve resistance to nuclease digestion in vivo.

[0710] While certain compounds, compositions, and methods of the present invention have been specifically described according to certain embodiments, the following examples are for illustrative purposes only and are not intended to limit the scope of the invention.

[0711] Example

[0712] Example 1: Exemplary synthesis of cationic lipids (4)

[0713] The cationic lipids described herein can be based on the following compounds. #4 The following exemplary synthesis is given for preparation:

[0714]

[0715]

[0716] #4

[0717] Amide synthesis (3)

[0718]

[0719] Amine (2) (1.00 g, 6.51 mmol) was added to a solution of acid (1) (5.14 g, 7.81 mmol), 4-dimethylaminopyridine (DMAP) (1.60 g, 13.02 mmol), and 1-ethyl-3-(3-dimethylaminopropyl)carbodiimide (EDC) (2.50 g, 13.02 mmol) in dichloromethane (DCM). The resulting mixture was stirred overnight at room temperature. After 16 h, MS analysis indicated the reaction was complete. The reaction mixture was diluted with DCM and washed with saturated sodium bicarbonate (NaHCO3) solution, water, and aqueous brine. The organic layer was dried over anhydrous sodium sulfate (Na2SO4) and concentrated. The crude residue was purified by column chromatography using an 80 g silica gel column, and the desired product was eluted with 9% ethyl acetate in hexane. The purest fraction was concentrated to obtain 3.71 g (75.3%) of pure product.

[0720] result:

[0721] ESI-MS analysis: Calculated value C 41 H 84 N2O4SSi2, [M + H + = 757.58, Observed value = 757.5

[0722] Amide deprotection (4)

[0723]

[0724] Hydrogen fluoride (70% HF.py complex, 4.41 mL, 48.99 mmol) was added to a solution of amide 3 (3.71 g, 4.90 mmol) in tetrahydrofuran (12 mL) at 0°C, and the mixture was stirred at the same temperature for 10 min. The reaction mixture was then warmed to room temperature and stirred for 16 h. MS analysis indicated that the reaction was complete. The reaction mixture was diluted with ethyl acetate and quenched by the slow addition of solid NaHCO3 at 0°C followed by the addition of a saturated NaHCO3 solution. The organic layer was washed with a saturated NaHCO3 solution, water, and brine. The resulting solution was then dried over anhydrous Na2SO4 and concentrated. The crude residue was purified, and the desired product was eluted with 6% methanol in DCM. The fraction containing the purified product was concentrated to obtain 2.07 g (79.9%) of pure product.

[0725] result:

[0726] ESI-MS analysis: Calculated value C 29 H 56 N₂O₄S, [M + H] + [ ] = 529.40, Observed value = 529.3

[0727] Thiol solution synthesis(8)

[0728]

[0729] Trifluoroacetic acid (TFA) (5.40 g, 47.38 mmol) was added to a solution of S-triphenylmethyl compound (7) (0.79 g, 1.02 mmol) in DCM (4 mL), and the mixture was stirred at room temperature for 30 min. Triethylsilane (0.14 g, 1.18 mmol) was then slowly added, and the mixture was stirred at room temperature for one hour. MS analysis indicated that the reaction was complete. The reaction mixture was concentrated via a rotary evaporator and dissolved in chloroform (12 mL) for later use.

[0730] result:

[0731] ESI-MS analysis: Calculated value C 32 H 67 NO2S, [M + H + = 530.50, Observed value = 530.5

[0732] Synthesis of R-SS-Py solution (6)

[0733]

[0734] A solution of thiolactone (4) (0.59 g, 1.116 mmol) in chloroform (20 mL) was added to dipyridyl disulfide (0.74 g, 3.35 mmol) and triethylamine (TEA) (0.640 g, 3.35 mmol), and stirred for 1 hour. Then, amine (5) (0.84 g, 6.70 mmol) was added and stirred overnight at room temperature. MS analysis indicated the reaction was complete. The reaction mixture was diluted with DCM and washed with saturated ammonium chloride solution, water, and aqueous salt solution. The organic layer was dried over anhydrous sodium sulfate (Na₂SO₄) and concentrated. The crude residue was purified by column chromatography using a 12 g silica gel column, and the desired product was eluted with 7% methanol in DCM. The purest fraction was concentrated to obtain 0.72 g (84.6%) of pure product.

[0735] result:

[0736] ESI-MS analysis: Calculated value C 40 H 70 N6O4S2, [M + H + = 763.50, Observed value = 763.5

[0737] Synthesis of thiolactone final products (9)

[0738] ≠4

[0739] A solution of thiol (8) in chloroform (8 mL) was added to a solution of R-SS-Py (6) (0.72 g, 0.943 mmol) in chloroform (10 mL) at room temperature. The resulting reaction mixture was stirred overnight at room temperature for 16 h. MS analysis indicated that the reaction was complete. The reaction mixture was concentrated. The crude residue was purified by column chromatography using a 24 g column, and the desired product was eluted with 16% MeOH in DCM. The fraction containing the product was concentrated to obtain 0.61 g (54.9%) of pure product.

[0740] result:

[0741] ESI-MS: Calculated value C 67 H 132 N6O6S2, [M + H + [M / 2 + H] = 1181.97, observed value = 1181.8, + ] = 591.5, and [M / 3 + H + = 394.9

[0742] The following compounds (Table 1) were prepared by applying and / or modifying the above synthetic procedures:

[0743]

[0744]

[0745]

[0746]

[0747]

[0748]

[0749]

[0750]

[0751]

[0752]

[0753]

[0754]

[0755]

[0756]

[0757]

[0758]

[0759]

[0760]

[0761]

[0762]

[0763]

[0764]

[0765]

[0766]

[0767]

[0768]

[0769]

[0770]

[0771] Yield and analytical data are summarized in Table 2 below:

[0772]

[0773]

[0774]

[0775]

[0776]

[0777]

[0778]

[0779]

[0780]

[0781] Example 2: Lipid nanoparticle formulation

[0782] 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 (which is hereby incorporated herein by reference in its entirety).

[0783] The lipid nanoparticles in this invention were formulated using Method A of WO 2018 / 089801 (see, for example, Example 1 and Figure 1 of WO 2018 / 089801). Method A (“A”) relates to a conventional method of encapsulating mRNA by mixing mRNA with a lipid mixture without pre-forming lipids into lipid nanoparticles. In one exemplary method, an ethanolic solution of a lipid mixture (cationic lipids, phosphatidylethanolamine, cholesterol, and polyethylene glycol-lipids) is combined with an aqueous buffer solution of the target mRNA at a fixed lipid-to-mRNA ratio under controlled conditions at an acidic pH to produce a homogeneous suspension of LNPs. After ultrafiltration and percolation into a suitable dilution system, the resulting nanoparticle suspension is diluted to the final concentration, filtered, and frozen at -80°C until use.

[0784] The lipid nanoparticle formulations in Table X were prepared using method A. All lipid nanoparticle formulations contained hEPO mRNA and different lipids (cationic lipids: DMG-PEG2000: cholesterol: DOPE / DSPC) in the mol% ratio specified in Table 3.

[0785] Table 3: Characterization of exemplary lipid nanoparticles

[0786]

[0787] The N / P ratio is defined as the ratio of the amount of nitrogen in cationic lipids to the amount of phosphate in nucleic acids.

[0788] The cationic lipids of the present invention were evaluated using lipid nanoparticle formulation 1. MC3 was evaluated using lipid nanoparticle formulation 2, which is a typical formulation of this lipid.

[0789] Example 3. Delivery of hEPO mRNA via intramuscular administration

[0790] In summary, lipid screening was conducted using 6-8 week old female BALB / cJ mice (Jax #000651). Mice were administered 0.1 µg of LNP in 30 µL via a single intramuscular (IM) injection into the gastrocnemius leg muscle. Blood samples were collected at 6 and 24 hours post-injection, and serum hEPO levels were measured using an ELISA assay according to the manufacturer's protocol (R&D Systems, catalog number DEP-00). WO 2022 / 099003 A1 also describes an in vivo assay for intramuscular administration (e.g., page 46, paragraph

[00206] ). The measured serum hEPO expression data are presented in Table 4 below. These data demonstrate that the disclosed lipids can induce the expression of the delivered mRNA.

[0791] Table 4. hEPO expression

[0792]

[0793] .

Claims

1. A compound having formula (I): (I) in m is an integer selected from 1 to 6; preferably 2 to 4; n is an integer selected from 1 to 6; preferably 2 to 4; p is an integer selected from 1 to 6; preferably 2 to 4; Same or different R 1 and R 2 Independently selected from the group consisting of: straight-chain or branched (C1-C30) alkyl and straight-chain or branched (C2-C30) alkenyl. Each of the alkyl and alkenyl groups is optionally separated by one or more groups selected from the following: –C=O-, -C=OO-, and –O-, and / or Each of the alkyl and alkenyl groups is optionally substituted by one or more substituents selected from the following: –OR, -CN-, -(C1-C6)alkyl-OH, -CF3, -NO2, -COOR, -SR, halogen atom and -NRR'; R 3 The group consisting of (C1-C6) alkyl groups selected from H, optionally substituted with one or more of the following substituents: –OR, -CN-, -(C1-C6)alkyl-OH, -CF3, -NO2, -COOR, -SR, halogen atom and -NRR'; Same or different R 4 and R 5 Independently selected from the group consisting of: straight-chain or branched (C1-C6) alkyl and straight-chain or branched (C2-C6) alkenyl, each alkyl or alkenyl optionally substituted by one or more substituents selected from the group consisting of: –OR, -CN-, -(C1-C6)alkyl-OH, -CF3, -NO2, -COOR, -SR, halogen atom and -NRR'; Or R 4 and R 5 Together with the N atoms to which they are attached, they form the following: Five- to six-membered cycloalkyl or heterocyclic compounds containing one to four heteroatoms selected from O, N, and S, or Five- to six-membered aryl or heteroaryl groups containing one to four heteroatoms selected from O, N, and S. The cycloalkyl, heterocyclic, aryl, or heteroaryl groups may optionally be substituted by one or more substituents selected from the following: -(C1-C6)alkyl, –OR, -CN-, -(C1-C6)alkyl-OH, -CF3, -NO2, -COOR, -SR, halogen atom, and -NRR'; Same or different R 6 and R 7 Independently selected from the group consisting of: straight-chain or branched (C1-C30) alkyl and straight-chain or branched (C2-C30) alkenyl. Each of the alkyl and alkenyl groups is optionally separated by one or more groups selected from the following: –C=O-, -C=OO-, and –O-, and / or Each of the alkyl and alkenyl groups is optionally substituted by one or more substituents selected from the following: –OR, -CN-, -C1-C6 alkyl-OH, -CF3, -NO2, -COOR, -SR, halogen atom and -NRR'; Same or different R 8 and R 9 Independently select the group composed of H and -OR; --- indicates an optional single bond, provided that, when present, compound (I) behaves as if formed by X. - The positive charge that counteracts the ion balance + The form represents the quaternary ammonium form, and when it is absent, compound (I) is in a neutral form and X is absent; X - These are anions that can be present at will; R 10 Choose from the group consisting of H and straight-chain or branched (C1-C30) alkyl groups; The same or different R and R' are independently selected from H and (C1-C6) alkyl groups; Or its pharmaceutically acceptable salts or esters.

2. A compound having formula (I) according to any one of the preceding claims, wherein Same or different R 1 and R 2 Independently selected from the group consisting of: straight-chain or branched (C1-C30) alkyl and straight-chain or branched (C2-C30) alkenyl. Each of the alkyl and alkenyl groups is optionally separated by one or more groups selected from –C=O- and –O-, and / or Each of the alkyl and alkenyl groups is optionally substituted by one or more substituents selected from –OH.

3. The compound having formula (I) according to claim 1 or 2, wherein the same or different R 6 and R 7 Independently selected from the group consisting of: straight-chain or branched (C1-C30) alkyl and straight-chain or branched (C2-C30) alkenyl. Each of the alkyl and alkenyl groups is optionally separated by one or more groups selected from –C=O- and –O-.

4. The compound having formula (I) according to claim 1, 2 or 3, wherein R 3 Select from the group consisting of H and (C1-C6) alkyl groups; Same or different R 4 and R 5 The group consisting of straight-chain or branched (C1-C6) alkyl groups is independently selected from the group consisting of one or more substituents selected from the group consisting of -OH. Or R 4 and R 5 Together with the N atoms to which they are attached, they form the following: Five- to six-membered cycloalkyl or heterocyclic compounds containing one to two heteroatoms selected from O, N, and S. Five- to six-membered aryl or heteroaryl groups containing one to two heteroatoms selected from O, N, and S. The cycloalkyl, heterocyclic, aryl, or heteroaryl groups may optionally be substituted by one or more substituents selected from the following: OH, -(C1-C6)alkyl, and C1-C6alkyl-OH.

5. A compound having formula (I) according to any one of the preceding claims, wherein X - Selected from Cl - ,Br - F - I - Trifluoroacetate, formate, p-toluenesulfonate, acetate, succinate, chlorate.

6. A compound having formula (I) according to any one of the preceding claims, wherein R 1 R 2 R 6 and R 7 Each of the following groups is independently selected: 。 7. A compound having formula (I) according to any one of the preceding claims, wherein R 4 and R 5 Independently selected from one of the following groups: Or they can form any of the following rings together with the N atoms to which they are attached: The premise is that any of the above rings may optionally be substituted by one or more substituents selected from: -–OR, -CN-, -(C1-C6)alkyl-OH, -CF3, -NO2, -COOR, -SR, halogen atom and -NRR', wherein R and R' are as defined in any one of claims 1 to 6.

8. A compound having formula (I) according to any one of the preceding claims, wherein the compound has formula (IA), (I-A'), (I-A''), (IB), (IC), (ID), (IE), (IF), (IG), (IH), (II), or (IJ): (I-A) (I-A’) (I-A’’) (I-B) (IC) (ID) (IE) (I-F) (I-G) (I-H) (I-I) (IJ) Where m, n, p, R 1 R 2 R 3 R 4 R 5 R 6 R 7 R 8 R 9 、- - -、R、R' and optional X、R 10 As defined in any one of claims 1 to 4; Furthermore, the same or different q and q' independently represent integers equal to 7, 8, 9, 10, or 11; Or its pharmaceutically acceptable salts or esters.

9. A compound having formula (I) according to any one of the preceding claims, wherein the compound is selected from the group consisting of: 18-((2-(diethylamino)ethyl)(methyl)carbamoyl)-7,26-dihydroxy-9-(2-hydroxy-8-(((Z)-non-2-en-1-yl)oxy)-8-oxooctyl)-24-(2-hydroxydodecyl)-20-oxo-14,15-dithia-9,19,24-triazahexadecanoic acid(Z)-non-2-en-1-yl ester 18-((2-(diethylamino)ethyl)(methyl)carbamoyl)-7,26-dihydroxy-9-(2-hydroxy-8-(((Z)-non-2-en-1-yl)oxy)-8-oxooctyl)-24-(2-hydroxydodecyl)-20-oxo-14,15-dithia-9,19,24-triazahexadecanoic acid(Z)-non-2-en-1-yl ester 18-((2-(diethylamino)ethyl)carbamoyl)-7,26-dihydroxy-9-(2-hydroxy-8-(((Z)-non-2-en-1-yl)oxy)-8-oxooctyl)-24-(2-hydroxydodecyl)-20-oxo-14,15-dithia-9,19,24-triazahexadecanoic acid(Z)-non-2-en-1-yl ester N-(1-((3-(1H-imidazol-1-yl)propyl)amino)-4-((4-(bis(2-hydroxytetradecyl)amino)butyl)dithioalkyl)-1-oxobutyl-2-yl)-5-(bis(2-hydroxydecyl)amino)pentanamide) 18-((3-(diethylamino)propyl)carbamoyl)-7,26-dihydroxy-9-(2-hydroxy-8-(((Z)-non-2-en-1-yl)oxy)-8-oxooctyl)-24-(2-hydroxydodecyl)-20-oxo-14,15-dithia-9,19,24-triazahexadecanoic acid(Z)-non-2-en-1-yl ester 18-((2-(dimethylamino)ethyl)carbamoyl)-7,26-dihydroxy-9-(2-hydroxy-8-(((Z)-non-2-en-1-yl)oxy)-8-oxooctyl)-24-(2-hydroxydodecyl)-20-oxo-14,15-dithia-9,19,24-triazahexadecanoic acid(Z)-non-2-en-1-yl ester 18-((2-(dimethylamino)ethyl)(methyl)carbamoyl)-7,26-dihydroxy-9-(2-hydroxy-8-(((Z)-non-2-en-1-yl)oxy)-8-oxooctyl)-24-(2-hydroxydodecyl)-20-oxo-14,15-dithia-9,19,24-triazahexadecanoic acid(Z)-non-2-en-1-yl ester 18-((3-(1H-imidazol-1-yl)propyl)carbamoyl)-7,26-dihydroxy-9-(2-hydroxy-8-(((Z)-non-2-en-1-yl)oxy)-8-oxooctyl)-24-(2-hydroxydodecyl)-20-oxo-14,15-dithia-9,19,24-triazahexahexadecanoic acid (Z)-non-2-en-1-yl ester 7,26-Dihydroxy-9-(2-hydroxy-8-(((Z)-non-2-en-1-yl)oxy)-8-oxooctyl)-24-(2-hydroxydodecyl)-20-oxo-18-((2-(piperidin-1-yl)ethyl)carbamoyl)-14,15-dithia-9,19,24-triazahexahexadecanoic acid(Z)-non-2-en-1-yl ester 7,26-Dihydroxy-9-(2-hydroxy-8-(((Z)-non-2-en-1-yl)oxy)-8-oxooctyl)-24-(2-hydroxydodecyl)-18-((4-morpholinobutyl)carbamoyl)-20-oxo-14,15-dithia-9,19,24-triazahexahexadecanoic acid (Z)-non-2-en-1-yl ester 7,25-Dihydroxy-9-(2-hydroxy-8-(((Z)-non-2-en-1-yl)oxy)-8-oxooctyl)-23-(2-hydroxydodecyl)-19-oxo-17-((4-(pyrrolidone-1-yl)butyl)carbamoyl)-14-thia-9,18,23-triazapentadecanoic acid (Z)-non-2-en-1-yl ester 5-(bis(2-hydroxydecyl)amino)-N-(19-hydroxy-17-(2-hydroxytetradecyl)-2-methyl-7-oxo-11,12-dithia-2,6,17-triazatrionecan-8-yl)pentanamide 5-(bis(2-hydroxydecyl)amino)-N-(3-ethyl-19-hydroxy-17-(2-hydroxytetradecyl)-7-oxo-11,12-dithia-3,6,17-triazatrionecan-8-yl)pentanamide 5-(bis(2-hydroxydecyl)amino)-N-(3-ethyl-20-hydroxy-18-(2-hydroxytetradecyl)-8-oxo-12,13-dithia-3,7,18-triazatridodecane-9-yl)pentanamide 5-(bis(2-hydroxydecyl)amino)-N-(18-hydroxy-16-(2-hydroxytetradecyl)-2-methyl-6-oxo-10,11-dithia-2,5,16-triazatriacontane-7-yl)pentanamide 5-(bis(2-hydroxydecyl)amino)-N-(18-hydroxy-16-(2-hydroxytetradecyl)-2,5-dimethyl-6-oxo-10,11-dithia-2,5,16-triazatriacontane-7-yl)pentanamide 5-(bis(2-hydroxydecyl)amino)-N-(3-ethyl-19-hydroxy-17-(2-hydroxytetradecyl)-6-methyl-7-oxo-11,12-dithia-3,6,17-triazatrionecan-8-yl)pentanamide 5-(bis(2-hydroxydecyl)amino)-N-(1,19-dihydroxy-17-(2-hydroxytetradecyl)-3-methyl-7-oxo-11,12-dithia-3,6,17-triazatrionecan-8-yl)pentanamide 5-(bis(2-hydroxydecyl)amino)-N-(4-((4-(bis(2-hydroxytetradecyl)amino)butyl)dithioalkyl)-1-((4-morpholinobutyl)amino)-1-oxobutyl-2-yl)pentanamide 5-(bis(2-hydroxydecyl)amino)-N-(4-((4-(bis(2-hydroxytetradecyl)amino)butyl)dithioalkyl)-1-oxo-1-((4-(pyrrolidone-1-yl)butyl)amino)but-2-yl)pentanamide 4-((4-(bis((9Z,12Z)-2-hydroxyoctadec-9,12-dien-1-yl)amino)butyl)dithioalkyl)-2-(4-(bis(2-hydroxydodecyl)amino)butamido)-N-(3-(dimethylamino)propyl)butamido) 4-((4-(bis((9Z,12Z)-2-hydroxyoctadec-9,12-dien-1-yl)amino)butyl)dithioalkyl)-2-(4-(bis(2-hydroxydodecyl)amino)butamido)-N-(2-(diethylamino)ethyl)butamido) 4-((4-(bis((9Z,12Z)-2-hydroxyoctadec-9,12-dien-1-yl)amino)butyl)dithioalkyl)-2-(4-(bis(2-hydroxydodecyl)amino)butamido)-N-(3-(diethylamino)propyl)butamido) 4-((4-(bis((9Z,12Z)-2-hydroxyoctadec-9,12-dien-1-yl)amino)butyl)dithioalkyl)-2-(4-(bis(2-hydroxydodecyl)amino)butamido)-N-(2-(dimethylamino)ethyl)butamido) 4-((4-(bis((9Z,12Z)-2-hydroxyoctadec-9,12-dien-1-yl)amino)butyl)dithioalkyl)-2-(4-(bis(2-hydroxydodecyl)amino)butamido)-N-(2-(dimethylamino)ethyl)-N-methylbutamido) 4-((4-(bis((9Z,12Z)-2-hydroxyoctadec-9,12-dien-1-yl)amino)butyl)dithioalkyl)-2-(4-(bis(2-hydroxydodecyl)amino)butamido)-N-(2-(diethylamino)ethyl)-N-methylbutamido) N-(3-(1H-imidazol-1-yl)propyl)-4-((4-(bis((9Z,12Z)-2-hydroxyoctadec-9,12-dien-1-yl)amino)butyl)dithioalkyl)-2-(4-(bis(2-hydroxydodecyl)amino)butyramide)butyramide 4-((4-(bis((9Z,12Z)-2-hydroxyoctadec-9,12-dien-1-yl)amino)butyl)dithioalkyl)-2-(4-(bis(2-hydroxydodecyl)amino)butamido)-N-(4-morpholinobutyl)butamido) 4-((4-(bis((9Z,12Z)-2-hydroxyoctadec-9,12-dien-1-yl)amino)butyl)dithioalkyl)-2-(4-(bis(2-hydroxydodecyl)amino)butamido)-N-(4-(pyrrolidone-1-yl)butyl)butamido) (Z)-18-((3-(dimethylamino)propyl)carbamoyl)-26-hydroxy-24-(2-hydroxydodecyl)-9-(octadec-9-en-1-yl)-20-oxo-14,15-dithia-9,19,24-triazahexadecanoic acid heptadecanoyl ester (Z)-18-((2-(diethylamino)ethyl)carbamoyl)-26-hydroxy-24-(2-hydroxydodecyl)-9-(octadec-9-en-1-yl)-20-oxo-14,15-dithia-9,19,24-triazahexadecanoic acid heptadecanoyl ester (Z)-18-((3-(diethylamino)propyl)carbamoyl)-26-hydroxy-24-(2-hydroxydodecyl)-9-(octadec-9-en-1-yl)-20-oxo-14,15-dithia-9,19,24-triazahexadecanoic acid heptadecanoyl ester (Z)-18-((2-(dimethylamino)ethyl)carbamoyl)-26-hydroxy-24-(2-hydroxydodecyl)-9-(octadec-9-en-1-yl)-20-oxo-14,15-dithia-9,19,24-triazahexadecanoic acid heptadecanoyl ester (Z)-18-((2-(dimethylamino)ethyl)(methyl)carbamoyl)-26-hydroxy-24-(2-hydroxydodecyl)-9-(octadec-9-en-1-yl)-20-oxo-14,15-dithia-9,19,24-triazahexadecanoic acid heptadecanoyl ester (Z)-18-((2-(diethylamino)ethyl)(methyl)carbamoyl)-26-hydroxy-24-(2-hydroxydodecyl)-9-(octadec-9-en-1-yl)-20-oxo-14,15-dithia-9,19,24-triazahexadecanoic acid heptadecanoyl ester (Z)-18-((3-(1H-imidazol-1-yl)propyl)carbamoyl)-26-hydroxy-24-(2-hydroxydodecyl)-9-(octadec-9-en-1-yl)-20-oxo-14,15-dithia-9,19,24-triazahexadecanoic acid heptadecanyl ester (Z)-26-hydroxy-24-(2-hydroxydodecyl)-18-((2-((2-hydroxyethyl)(methyl)amino)ethyl)carbamoyl)-9-(octadec-9-en-1-yl)-20-oxo-14,15-dithia-9,19,24-triazahexadecanoic acid heptadecanyl ester (Z)-26-hydroxy-24-(2-hydroxydodecyl)-18-((4-morpholinobutyl)carbamoyl)-9-(octadec-9-en-1-yl)-20-oxo-14,15-dithia-9,19,24-triazahexadecanoic acid heptadecanoyl ester (Z)-26-hydroxy-24-(2-hydroxydodecyl)-9-(octadec-9-en-1-yl)-20-oxo-18-((4-(pyrrolidin-1-yl)butyl)carbamoyl)-14,15-dithia-9,19,24-triazahexadecanoic acid heptadecanoyl ester 18-((3-(dimethylamino)propyl)carbamoyl)-25-hydroxy-23-(2-hydroxydecyl)-9-(8-(((Z)-non-2-en-1-yl)oxy)-8-oxooctyl)-20-oxo-14,15-dithia-9,19,23-triazatriacontaneanoic acid (Z)-non-2-en-1-yl ester 18-((2-(diethylamino)ethyl)carbamoyl)-25-hydroxy-23-(2-hydroxydecyl)-9-(8-(((Z)-non-2-en-1-yl)oxy)-8-oxooctyl)-20-oxo-14,15-dithia-9,19,23-triazatriacontaneanoic acid (Z)-non-2-en-1-yl ester 18-((3-(diethylamino)propyl)carbamoyl)-25-hydroxy-23-(2-hydroxydecyl)-9-(8-(((Z)-non-2-en-1-yl)oxy)-8-oxooctyl)-20-oxo-14,15-dithia-9,19,23-triazatriacontaneanoic acid (Z)-non-2-en-1-yl ester 18-((2-(dimethylamino)ethyl)carbamoyl)-25-hydroxy-23-(2-hydroxydecyl)-9-(8-(((Z)-non-2-en-1-yl)oxy)-8-oxooctyl)-20-oxo-14,15-dithia-9,19,23-triazatriacontaneanoic acid (Z)-non-2-en-1-yl ester 18-((2-(dimethylamino)ethyl)(methyl)carbamoyl)-25-hydroxy-23-(2-hydroxydecyl)-9-(8-(((Z)-non-2-en-1-yl)oxy)-8-oxooctyl)-20-oxo-14,15-dithia-9,19,23-triazatriacontaracanoic acid (Z)-non-2-en-1-yl ester 18-((2-(diethylamino)ethyl)(methyl)carbamoyl)-25-hydroxy-23-(2-hydroxydecyl)-9-(8-(((Z)-non-2-en-1-yl)oxy)-8-oxooctyl)-20-oxo-14,15-dithia-9,19,23-triazatriacontaneanoic acid (Z)-non-2-en-1-yl ester 18-((3-(1H-imidazol-1-yl)propyl)carbamoyl)-25-hydroxy-23-(2-hydroxydecyl)-9-(8-(((Z)-non-2-en-1-yl)oxy)-8-oxooctyl)-20-oxo-14,15-dithia-9,19,23-triazatriacontaneanoic acid (Z)-non-2-en-1-yl ester 25-Hydroxy-23-(2-Hydroxydecyl)-9-(8-(((Z)-Non-2-en-1-yl)oxy)-8-oxooctyl)-20-oxo-18-((2-(piperidin-1-yl)ethyl)glycyl)-14,15-dithia-9,19,23-triazatriacontaracanoic acid (Z)-non-2-en-1-yl ester 25-Hydroxy-23-(2-Hydroxydecyl)-18-((4-morpholinobutyl)carbamoyl)-9-(8-(((Z)-non-2-en-1-yl)oxy)-8-oxooctyl)-20-oxo-14,15-dithia-9,19,23-triazatriacontaracanoic acid (Z)-non-2-en-1-yl ester 25-Hydroxy-23-(2-Hydroxydecyl)-9-(8-(((Z)-Non-2-en-1-yl)oxy)-8-oxooctyl)-20-oxo-18-((4-(pyrrolidone-1-yl)butyl)carbamoyl)-14,15-dithia-9,19,23-triazatriacontaracanonicoic acid (Z)-non-2-en-1-yl ester 18-((3-(dimethylamino)propyl)carbamoyl)-26-hydroxy-24-(2-hydroxy-8-(((Z)-non-2-en-1-yl)oxy)-8-oxooctyl)-9-(8-(((Z)-non-2-en-1-yl)oxy)-8-oxooctyl)-20-oxo-14,15-dithia-9,19,24-triazatridodecanoic acid di((Z)-non-2-en-1-yl) ester 18-((2-(diethylamino)ethyl)carbamoyl)-26-hydroxy-24-(2-hydroxy-8-(((Z)-non-2-en-1-yl)oxy)-8-oxooctyl)-9-(8-(((Z)-non-2-en-1-yl)oxy)-8-oxooctyl)-20-oxo-14,15-dithia-9,19,24-triazatridodecanoic acid di((Z)-non-2-en-1-yl) ester 18-((3-(diethylamino)propyl)carbamoyl)-26-hydroxy-24-(2-hydroxy-8-(((Z)-non-2-en-1-yl)oxy)-8-oxooctyl)-9-(8-(((Z)-non-2-en-1-yl)oxy)-8-oxooctyl)-20-oxo-14,15-dithia-9,19,24-triazatridodecanoic acid di((Z)-non-2-en-1-yl) ester 18-((2-(dimethylamino)ethyl)carbamoyl)-26-hydroxy-24-(2-hydroxy-8-(((Z)-non-2-en-1-yl)oxy)-8-oxooctyl)-9-(8-(((Z)-non-2-en-1-yl)oxy)-8-oxooctyl)-20-oxo-14,15-dithia-9,19,24-triazatridodecanoic acid di((Z)-non-2-en-1-yl) ester 18-((2-(dimethylamino)ethyl)(methyl)carbamoyl)-26-hydroxy-24-(2-hydroxy-8-(((Z)-non-2-en-1-yl)oxy)-8-oxooctyl)-9-(8-(((Z)-non-2-en-1-yl)oxy)-8-oxooctyl)-20-oxo-14,15-dithia-9,19,24-triazatridodecanoic acid di((Z)-non-2-en-1-yl) ester 18-((2-(diethylamino)ethyl)(methyl)carbamoyl)-26-hydroxy-24-(2-hydroxy-8-(((Z)-non-2-en-1-yl)oxy)-8-oxooctyl)-9-(8-(((Z)-non-2-en-1-yl)oxy)-8-oxooctyl)-20-oxo-14,15-dithia-9,19,24-triazatridodecanoic acid di((Z)-non-2-en-1-yl) ester 18-((3-(1H-imidazol-1-yl)propyl)carbamoyl)-26-hydroxy-24-(2-hydroxy-8-(((Z)-non-2-en-1-yl)oxy)-8-oxooctyl)-9-(8-((((Z)-non-2-en-1-yl)oxy)-8-oxooctyl)-20-oxo-14,15-dithia-9,19,24-triazatridodecanoic acid di((Z)-non-2-en-1-yl) ester 26-Hydroxy-24-(2-hydroxy-8-(((Z)-non-2-en-1-yl)oxy)-8-oxooctyl)-9-(8-(((Z)-non-2-en-1-yl)oxy)-8-oxooctyl)-20-oxo-18-((2-(piperidin-1-yl)ethyl)glycyl)-14,15-dithia-9,19,24-triazatridodecanoic acid di((Z)-non-2-en-1-yl) ester 26-Hydroxy-24-(2-hydroxy-8-(((Z)-non-2-en-1-yl)oxy)-8-oxooctyl)-18-((4-morpholinobutyl)carbamoyl)-9-(8-(((Z)-non-2-en-1-yl)oxy)-8-oxooctyl)-20-oxo-14,15-dithia-9,19,24-triazatridodecanoic acid di((Z)-non-2-en-1-yl) ester 26-Hydroxy-24-(2-hydroxy-8-(((Z)-non-2-en-1-yl)oxy)-8-oxooctyl)-9-(8-(((Z)-non-2-en-1-yl)oxy)-8-oxooctyl)-20-oxo-18-((4-(pyrrolidone-1-yl)butyl)carbamoyl)-14,15-dithia-9,19,24-triazatridodecanoic acid di((Z)-non-2-en-1-yl) ester 18-((3-(dimethylamino)propyl)carbamoyl)-7,26-dihydroxy-9,24-bis(2-hydroxy-8-(((Z)-non-2-en-1-yl)oxy)-8-oxooctyl)-20-oxo-14,15-dithia-9,19,24-triazatridodecanoic acid di((Z)-non-2-en-1-yl) ester 18-((2-(diethylamino)ethyl)carbamoyl)-7,26-dihydroxy-9,24-bis(2-hydroxy-8-(((Z)-non-2-en-1-yl)oxy)-8-oxooctyl)-20-oxo-14,15-dithia-9,19,24-triazatridodecanoic acid di((Z)-non-2-en-1-yl) ester 18-((3-(dimethylamino)propyl)carbamoyl)-26-hydroxy-24-(2-hydroxydodecyl)-9-(8-(((Z)-non-2-en-1-yl)oxy)-8-oxooctyl)-20-oxo-14,15-dithia-9,19,24-triazahexadecanoic acid (Z)-non-2-en-1-yl ester 18-((2-(diethylamino)ethyl)carbamoyl)-26-hydroxy-24-(2-hydroxydodecyl)-9-(8-(((Z)-non-2-en-1-yl)oxy)-8-oxooctyl)-20-oxo-14,15-dithia-9,19,24-triazahexadecanoic acid (Z)-non-2-en-1-yl ester 18-((3-(diethylamino)propyl)carbamoyl)-26-hydroxy-24-(2-hydroxydodecyl)-9-(8-(((Z)-non-2-en-1-yl)oxy)-8-oxooctyl)-20-oxo-14,15-dithia-9,19,24-triazahexadecanoic acid (Z)-non-2-en-1-yl ester 18-((2-(dimethylamino)ethyl)carbamoyl)-26-hydroxy-24-(2-hydroxydodecyl)-9-(8-(((Z)-non-2-en-1-yl)oxy)-8-oxooctyl)-20-oxo-14,15-dithia-9,19,24-triazahexadecanoic acid (Z)-non-2-en-1-yl ester 18-((2-(dimethylamino)ethyl)(methyl)carbamoyl)-26-hydroxy-24-(2-hydroxydodecyl)-9-(8-(((Z)-non-2-en-1-yl)oxy)-8-oxooctyl)-20-oxo-14,15-dithia-9,19,24-triazahexadecanoic acid(Z)-non-2-en-1-yl ester 18-((2-(diethylamino)ethyl)(methyl)carbamoyl)-26-hydroxy-24-(2-hydroxydodecyl)-9-(8-(((Z)-non-2-en-1-yl)oxy)-8-oxooctyl)-20-oxo-14,15-dithia-9,19,24-triazahexadecanoic acid (Z)-non-2-en-1-yl ester 18-((3-(1H-imidazol-1-yl)propyl)carbamoyl)-26-hydroxy-24-(2-hydroxydodecyl)-9-(8-(((Z)-non-2-en-1-yl)oxy)-8-oxooctyl)-20-oxo-14,15-dithia-9,19,24-triazahexadecanoic acid(Z)-non-2-en-1-yl ester 26-Hydroxy-24-(2-Hydroxydodecyl)-9-(8-(((Z)-Non-2-en-1-yl)oxy)-8-oxooctyl)-20-oxo-18-((2-(piperidin-1-yl)ethyl)carbamoyl)-14,15-dithia-9,19,24-triazahexahexadecanoic acid (Z)-non-2-en-1-yl ester 26-Hydroxy-24-(2-Hydroxydodecyl)-18-((4-morpholinobutyl)carbamoyl)-9-(8-(((Z)-non-2-en-1-yl)oxy)-8-oxooctyl)-20-oxo-14,15-dithia-9,19,24-triazahexadecanoic acid (Z)-non-2-en-1-yl ester 26-Hydroxy-24-(2-Hydroxydodecyl)-9-(8-(((Z)-Non-2-en-1-yl)oxy)-8-oxooctyl)-20-oxo-18-((4-(pyrrolidone-1-yl)butyl)carbamoyl)-14,15-dithia-9,19,24-triazahexahexadecanoic acid (Z)-non-2-en-1-yl ester 5-(bis(2-hydroxydecyl)amino)-N-(19-hydroxy-17-(2-hydroxydodecyl)-2-methyl-7-oxo-11,12-dithia-2,6,17-triazanonadecan-8-yl)pentanamide 5-(bis(2-hydroxydecyl)amino)-N-(3-ethyl-19-hydroxy-17-(2-hydroxydodecyl)-7-oxo-11,12-dithia-3,6,17-triazanonadecan-8-yl)pentanamide 5-(bis(2-hydroxydecyl)amino)-N-(3-ethyl-20-hydroxy-18-(2-hydroxydodecyl)-8-oxo-12,13-dithia-3,7,18-triazatriacontane-9-yl)pentanamide 5-(bis(2-hydroxydecyl)amino)-N-(18-hydroxy-16-(2-hydroxydodecyl)-2-methyl-6-oxo-10,11-dithia-2,5,16-triazaoctacosane-7-yl)pentanamide 5-(bis(2-hydroxydecyl)amino)-N-(18-hydroxy-16-(2-hydroxydodecyl)-2,5-dimethyl-6-oxo-10,11-dithia-2,5,16-triazaoctacosane-7-yl)pentanamide 5-(bis(2-hydroxydecyl)amino)-N-(3-ethyl-19-hydroxy-17-(2-hydroxydodecyl)-6-methyl-7-oxo-11,12-dithia-3,6,17-triazanonadecan-8-yl)pentanamide N-(1-((3-(1H-imidazol-1-yl)propyl)amino)-4-((4-(bis(2-hydroxydodecyl)amino)butyl)dithioalkyl)-1-oxobutyl-2-yl)-5-(bis(2-hydroxydecyl)amino)pentanamide) 5-(bis(2-hydroxydecyl)amino)-N-(4-((4-(bis(2-hydroxydodecyl)amino)butyl)dithioalkyl)-1-oxo-1-((3-(piperidin-1-yl)propyl)amino)but-2-yl)pentanamide 5-(bis(2-hydroxydecyl)amino)-N-(4-((4-(bis(2-hydroxydodecyl)amino)butyl)dithioalkyl)-1-oxo-1-((2-(pyrrolidone-1-yl)ethyl)amino)but-2-yl)pentanamide 5-(bis(2-hydroxydecyl)amino)-N-(4-((4-(bis(2-hydroxydodecyl)amino)butyl)dithioalkyl)-1-oxo-1-((4-(pyrrolidone-1-yl)butyl)amino)but-2-yl)pentanamide 18-((3-(dimethylamino)propyl)carbamoyl)-25-hydroxy-23-(2-hydroxydecyl)-20-oxo-9-(6-oxo-6-(undecyloxy)hexyl)-14,15-dithia-9,19,23-triazatriacontane-octadecanoate-9-yl ester 18-((2-(diethylamino)ethyl)carbamoyl)-25-hydroxy-23-(2-hydroxydecyl)-20-oxo-9-(6-oxo-6-(undecyloxy)hexyl)-14,15-dithia-9,19,23-triazatriacontane-octadecanoate-9-yl ester 8-[4-[[3-[3-[bis(2-hydroxydecyl)amino]propionylamino]-4-[3-(diethylamino)propylamino]-4-oxo-butyl]dithioalkyl]butyl-(6-oxo-6-undecoxy-hexyl)amino]octanoic acid 1-octylnonyl ester 8-[4-[[3-[3-[bis(2-hydroxydecyl)amino]propionylamino]-4-[2-(dimethylamino)ethylamino]-4-oxo-butyl]dithioalkyl]butyl-(6-oxo-6-undecoxy-hexyl)amino]octanoic acid 1-octylnonyl ester 8-[4-[[3-[3-[bis(2-hydroxydecyl)amino]propionylamino]-4-[2-(dimethylamino)ethyl-methyl-amino]-4-oxo-butyl]dithioalkyl]butyl-(6-oxo-6-undecoxy-hexyl)amino]octanoic acid 1-octylnonyl ester 8-[4-[[3-[3-[bis(2-hydroxydecyl)amino]propionylamino]-4-[2-(diethylamino)ethyl-methyl-amino]-4-oxo-butyl]dithioalkyl]butyl-(6-oxo-6-undecoxy-hexyl)amino]octanoic acid 1-octylnonyl ester 8-[4-[[3-[3-[bis(2-hydroxydecyl)amino]propionylamino]-4-(3-imidazol-1-ylpropylamino)-4-oxo-butyl]dithioalkyl]butyl-(6-oxo-6-undecoxy-hexyl)amino]octanoic acid 1-octylnonyl ester 8-[4-[[3-[3-[bis(2-hydroxydecyl)amino]propionylamino]-4-(4-morpholinobutylamino)-4-oxo-butyl]dithioalkyl]butyl-(6-oxo-6-undecoxy-hexyl)amino]octanoic acid 1-octylnonyl ester 8-[4-[[3-[3-[bis(2-hydroxydecyl)amino]propionylamino]-4-oxo-4-(4-pyrrolidine-1-ylbutylamino)butyl]dithioalkyl]butyl-(6-oxo-6-undecoxy-hexyl)amino]octanoic acid 1-octylnonyl ester 8-[[4-[[3-[4-[bis(2-hydroxytetradecyl)amino]butyldithioalkyl]-1-[3-(dimethylamino)propylcarbamoyl]propyl]amino]-4-oxo-butyl]-[2-hydroxy-8-[(Z)-non-2-enoxy]-8-oxo-octyl]amino]-7-hydroxy-octanoic acid[(Z)-non-2-enyl] ester 8-[[4-[[3-[4-[bis(2-hydroxytetradecyl)amino]butyldithioalkyl]-1-[2-(diethylamino)ethylcarbamoyl]propyl]amino]-4-oxo-butyl]-[2-hydroxy-8-[(Z)-non-2-enoxy]-8-oxo-octyl]amino]-7-hydroxy-octanoic acid[(Z)-non-2-enyl] ester 8-[[4-[[3-[4-[bis(2-hydroxytetradecyl)amino]butyldithioalkyl]-1-[3-(diethylamino)propylcarbamoyl]propyl]amino]-4-oxo-butyl]-[2-hydroxy-8-[(Z)-non-2-enoxy]-8-oxo-octyl]amino]-7-hydroxy-octanoic acid[(Z)-non-2-enyl] ester 8-[[4-[[3-[4-[bis(2-hydroxytetradecyl)amino]butyldithioalkyl]-1-[2-(dimethylamino)ethylcarbamoyl]propyl]amino]-4-oxo-butyl]-[2-hydroxy-8-[(Z)-non-2-enoxy]-8-oxo-octyl]amino]-7-hydroxy-octanoic acid[(Z)-non-2-enyl] ester 8-[[4-[[3-[4-[bis(2-hydroxytetradecyl)amino]butyldithioalkyl]-1-[2-(dimethylamino)ethyl-methyl-carbamoyl]propyl]amino]-4-oxo-butyl]-[2-hydroxy-8-[(Z)-non-2-enoxy]-8-oxo-octyl]amino]-7-hydroxy-octanoic acid[(Z)-non-2-enyl] ester 8-[[4-[[3-[4-[bis(2-hydroxytetradecyl)amino]butyldithioalkyl]-1-[2-(diethylamino)ethyl-methyl-carbamoyl]propyl]amino]-4-oxo-butyl]-[2-hydroxy-8-[(Z)-non-2-enoxy]-8-oxo-octyl]amino]-7-hydroxy-octanoic acid[(Z)-non-2-enyl] ester 8-[[4-[[3-[4-[bis(2-hydroxytetradecyl)amino]butyldithioalkyl]-1-(3-imidazol-1-ylpropylcarbamoyl)propyl]amino]-4-oxo-butyl]-[2-hydroxy-8-[(Z)-non-2-enoxy]-8-oxo-octyl]amino]-7-hydroxy-octanoic acid[(Z)-non-2-enyl] ester 8-[[4-[[3-[4-[bis(2-hydroxytetradecyl)amino]butyldithioalkyl]-1-[2-(1-piperidinyl)ethylcarbamoyl]propyl]amino]-4-oxo-butyl]-[2-hydroxy-8-[(Z)-non-2-enoxy]-8-oxo-octyl]amino]-7-hydroxy-octanoic acid[(Z)-non-2-enyl] ester 8-[[4-[[3-[4-[bis(2-hydroxytetradecyl)amino]butyldithioalkyl]-1-(4-morpholinobutylcarbamoyl)propyl]amino]-4-oxo-butyl]-[2-hydroxy-8-[(Z)-non-2-enoxy]-8-oxo-octyl]amino]-7-hydroxy-octanoic acid[(Z)-non-2-enyl] ester 8-[[4-[[3-[4-[bis(2-hydroxytetradecyl)amino]butyldithioalkyl]-1-(4-pyrrolidine-1-ylbutylcarbamoyl)propyl]amino]-4-oxo-butyl]-[2-hydroxy-8-[(Z)-non-2-enoxy]-8-oxo-octyl]amino]-7-hydroxy-octanoic acid[(Z)-non-2-enyl] ester 8-[[4-[[1-[3-(dimethylamino)propylcarbamoyl]-3-[4-[[8-(1-octylnonoxy)-8-oxo-octyl]-(6-oxo-6-undecoxy-hexyl)amino]butyldithioyl]propyl]amino]-4-oxo-butyl]-[2-hydroxy-8-[(Z)-non-1-enoxy]-8-oxo-octyl]amino]-7-hydroxy-octanoic acid[(Z)-non-1-enyl] ester 8-[[4-[[1-[2-(diethylamino)ethylcarbamoyl]-3-[4-[[8-(1-octylnonoxy)-8-oxo-octyl]-(6-oxo-6-undecoxy-hexyl)amino]butyldithioyl]propyl]amino]-4-oxo-butyl]-[2-hydroxy-8-[(Z)-non-2-enoxy]-8-oxo-octyl]amino]-7-hydroxy-octanoic acid[(Z)-non-2-enyl] ester] 8-[[4-[[1-[3-(diethylamino)propylcarbamoyl]-3-[4-[[8-(1-octylnonoxy)-8-oxo-octyl]-(6-oxo-6-undecoxy-hexyl)amino]butyldithioyl]propyl]amino]-4-oxo-butyl]-[2-hydroxy-8-[(Z)-non-1-enoxy]-8-oxo-octyl]amino]-7-hydroxy-octanoic acid[(Z)-non-1-enyl] ester] 18-((2-(dimethylamino)ethyl)carbamoyl)-26-hydroxy-24-(2-hydroxy-8-(((Z)-non-2-en-1-yl)oxy)-8-oxooctyl)-20-oxo-9-(6-oxo-6-(undecyloxy)hexyl)-14,15-dithia-9,19,24-triazatridodecanoic acid 1-(heptadecano-9-yl)32-((Z)-non-2-en-1-yl) ester 8-[[4-[[1-[2-(dimethylamino)ethyl-methyl-carbamoyl]-3-[4-[[8-(1-octylnonoxy)-8-oxo-octyl]-(6-oxo-6-undecoxy-hexyl)amino]butyldithioyl]propyl]amino]-4-oxo-butyl]-[2-hydroxy-8-[(Z)-non-2-enoxy]-8-oxo-octyl]amino]-7-hydroxy-octanoic acid[(Z)-non-2-enyl] ester] 8-[[4-[[1-[2-(diethylamino)ethyl-methyl-carbamoyl]-3-[4-[[8-(1-octylnonoxy)-8-oxo-octyl]-(6-oxo-6-undecoxy-hexyl)amino]butyldithioyl]propyl]amino]-4-oxo-butyl]-[2-hydroxy-8-[(Z)-non-2-enoxy]-8-oxo-octyl]amino]-7-hydroxy-octanoic acid[(Z)-non-2-enyl] ester] 7-Hydroxy-8-[[2-Hydroxy-8-[(Z)-Non-2-enoxy]-8-oxo-octyl]-[4-[[1-(3-imidazol-1-ylpropylcarbamoyl)-3-[4-[[8-(1-octylnonoxy)-8-oxo-octyl]-(6-oxo-6-undecoxy-hexyl)amino]butyldithioyl]propyl]amino]-4-oxo-butyl]amino]octanoic acid[(Z)-non-2-enyl] ester 7-Hydroxy-8-[[2-Hydroxy-8-[(Z)-Non-2-enoxy]-8-oxo-octyl]-[4-[[3-[4-[[8-(1-octylnonoxy)-8-oxo-octyl]-(6-oxo-6-undecoxy-hexyl)amino]butyldithioyl]-1-[2-(1-piperidinyl)ethylcarbamoyl]propyl]amino]-4-oxo-butyl]amino]octanoic acid[(Z)-non-2-enyl] ester 7-Hydroxy-8-[[2-Hydroxy-8-[(Z)-Non-2-enoxy]-8-oxo-octyl]-[4-[[1-(4-morpholinobutylcarbamoyl)-3-[4-[[8-(1-octylnonoxy)-8-oxo-octyl]-(6-oxo-6-undecoxy-hexyl)amino]butyldithioyl]propyl]amino]-4-oxo-butyl]amino]octanoic acid[(Z)-non-2-enyl] ester 7-Hydroxy-8-[[2-Hydroxy-8-[(Z)-Non-2-enoxy]-8-oxo-octyl]-[4-[[3-[4-[[8-(1-octylnonoxy)-8-oxo-octyl]-(6-oxo-6-undecoxy-hexyl)amino]butyldithioalkyl]-1-(4-pyrrolidine-1-ylbutylcarbamoyl)propyl]amino]-4-oxo-butyl]amino]octanoic acid[(Z)-non-2-enyl] ester 8-[4-[[3-[4-[bis(2-hydroxydodecyl)amino]butyrylamino]-4-[3-(dimethylamino)propylamino]-4-oxo-butyl]dithioalkyl]butyl-(6-oxo-6-undecoxy-hexyl)amino]octanoic acid 1-octylnonyl ester 8-[4-[[3-[4-[bis(2-hydroxydodecyl)amino]butyrylamino]-4-[2-(diethylamino)ethylamino]-4-oxo-butyl]dithioalkyl]butyl-(6-oxo-6-undecoxy-hexyl)amino]octanoic acid 1-octylnonyl ester 8-[4-[[3-[4-[bis(2-hydroxydodecyl)amino]butyrylamino]-4-[3-(diethylamino)propylamino]-4-oxo-butyl]dithioalkyl]butyl-(6-oxo-6-undecoxy-hexyl)amino]octanoic acid 1-octylnonyl ester 8-[4-[[3-[4-[bis(2-hydroxydodecyl)amino]butyrylamino]-4-[2-(dimethylamino)ethylamino]-4-oxo-butyl]dithioalkyl]butyl-(6-oxo-6-undecoxy-hexyl)amino]octanoic acid 1-octylnonyl ester 8-[4-[[3-[4-[bis(2-hydroxydodecyl)amino]butyrylamino]-4-[2-(dimethylamino)ethyl-methyl-amino]-4-oxo-butyl]dithioalkyl]butyl-(6-oxo-6-undecoxy-hexyl)amino]octanoic acid 1-octylnonyl ester 8-[4-[[3-[4-[bis(2-hydroxydodecyl)amino]butyrylamino]-4-[2-(diethylamino)ethyl-methyl-amino]-4-oxo-butyl]dithioalkyl]butyl-(6-oxo-6-undecoxy-hexyl)amino]octanoic acid 1-octylnonyl ester 8-[4-[[3-[4-[bis(2-hydroxydodecyl)amino]butyrylamino]-4-(3-imidazol-1-ylpropylamino)-4-oxo-butyl]dithioalkyl]butyl-(6-oxo-6-undecoxy-hexyl)amino]octanoic acid 1-octylnonyl ester 8-[4-[[3-[4-[bis(2-hydroxydodecyl)amino]butyrylamino]-4-oxo-4-[2-(1-piperidinyl)ethylamino]butyl]dithioalkyl]butyl-(6-oxo-6-undecoxy-hexyl)amino]octanoic acid 1-octylnonyl ester 8-[4-[[3-[4-[bis(2-hydroxydodecyl)amino]butyrylamino]-4-(4-morpholinobutylamino)-4-oxo-butyl]dithioalkyl]butyl-(6-oxo-6-undecoxy-hexyl)amino]octanoic acid 1-octylnonyl ester 8-[4-[[3-[4-[bis(2-hydroxydodecyl)amino]butyrylamino]-4-oxo-4-(4-pyrrolidine-1-ylbutylamino)butyl]dithioalkyl]butyl-(6-oxo-6-undecoxy-hexyl)amino]octanoic acid 1-octylnonyl ester 18-((3-(2,7a-dihydro-1H-benzo[d]imidazol-1-yl)propyl)carbamoyl)-27-hydroxy-25-(2-hydroxydecyl)-20-oxo-9-(6-oxo-6-(undecyloxy)hexyl)-14,15-dithia-9,19,25-triazapentadecanoic acid heptadecano-9-yl ester 18-((3-(1H-imidazol-1-yl)propyl)carbamoyl)-27-hydroxy-25-(2-hydroxydecyl)-20-oxo-9-(6-oxo-6-(undecyloxy)hexyl)-14,15-dithia-9,19,25-triazapentadecanoic acid heptadecano-9-yl ester N-(3-(1H-imidazol-1-yl)propyl)-4-((4-(bis(2-hydroxydecyl)amino)butyl)dithio)-2-(4-(bis(2-hydroxydodecyl)amino)butyramido)butyramide N-(1-((4-(1H-imidazol-1-yl)butyl)amino)-4-((4-(bis(2-hydroxytetradecyl)amino)butyl)dithioalkyl)-1-oxobutyl-2-yl)-5-(bis(2-hydroxydecyl)amino)pentanamide N-(1-((2-(1H-imidazol-1-yl)ethyl)amino)-4-((4-(bis(2-hydroxytetradecyl)amino)butyl)dithioalkyl)-1-oxobutyl-2-yl)-5-(bis(2-hydroxydecyl)amino)pentanamide 5-(bis(2-hydroxydecyl)amino)-N-(4-((4-(bis(2-hydroxytetradecyl)amino)butyl)dithioalkyl)-1-((3-(2-methyl-1H-imidazol-1-yl)propyl)amino)-1-oxobut-2-yl)pentanamide N-(1-((2-(1H-indol-3-yl)ethyl)amino)-4-((4-(bis(2-hydroxytetradecyl)amino)butyl)dithioalkyl)-1-oxobutyl-2-yl)-5-(bis(2-hydroxydecyl)amino)pentanamide) 2-[4-[bis(2-hydroxydodecyl)amino]butyrylamino]-4-[4-[bis[(Z)-2-hydroxyoctadec-9-enyl]amino]butyldithioalkyl]-N-[3-(dimethylamino)propyl]butyramide 2-[4-[bis(2-hydroxydodecyl)amino]butyrylamino]-4-[4-[bis[(Z)-2-hydroxyoctadec-9-enyl]amino]butyldithioalkyl]-N-[2-(diethylamino)ethyl]butyramide 2-[4-[bis(2-hydroxydodecyl)amino]butyrylamino]-4-[4-[bis[(Z)-2-hydroxyoctadec-9-enyl]amino]butyldithioalkyl]-N-[3-(diethylamino)propyl]butyramide 2-[4-[bis(2-hydroxydodecyl)amino]butyrylamino]-4-[4-[bis[(Z)-2-hydroxyoctadec-9-enyl]amino]butyldithioalkyl]-N-[2-(dimethylamino)ethyl]butyramide 2-[4-[bis(2-hydroxydodecyl)amino]butyrylamino]-4-[4-[bis[(Z)-2-hydroxyoctadec-9-enyl]amino]butyldithioalkyl]-N-[2-(dimethylamino)ethyl]-N-methylbutyramide; methane 2-[4-[bis(2-hydroxydodecyl)amino]butyrylamino]-4-[4-[bis[(Z)-2-hydroxyoctadec-9-enyl]amino]butyldithioalkyl]-N-[2-(diethylamino)ethyl]-N-methylbutyramide N-(3-(1H-imidazol-1-yl)propyl)-4-((4-(bis((Z)-2-hydroxyoctadec-9-en-1-yl)amino)butyl)dithioyl)-2-(4-(bis(2-hydroxydodecyl)amino)butyramido)butyramide 2-[4-[bis(2-hydroxydodecyl)amino]butyrylamino]-4-[4-[bis[(Z)-2-hydroxyoctadec-9-enyl]amino]butyldithioalkyl]-N-[3-(2-methylimidazol-1-yl)propyl]butyramide 2-[4-[bis(2-hydroxydodecyl)amino]butyrylamino]-4-[4-[bis[(Z)-2-hydroxyoctadec-9-enyl]amino]butyldithioalkyl]-N-(4-morpholinobutyl)butyramide 2-[4-[bis(2-hydroxydodecyl)amino]butyrylamino]-4-[4-[bis[(Z)-2-hydroxyoctadec-9-enyl]amino]butyldithioalkyl]-N-(4-pyrrolidine-1-ylbutyl)butyramide 4-((4-(bis(2-hydroxydecyl)amino)butyl)dithioalkyl)-2-(4-(bis(2-hydroxydodecyl)amino)butamido)-N-(3-(dimethylamino)propyl)butamido) 4-((4-(bis(2-hydroxydecyl)amino)butyl)dithioalkyl)-2-(4-(bis(2-hydroxydodecyl)amino)butamido)-N-(2-(diethylamino)ethyl)butamido) 4-((4-(bis(2-hydroxydecyl)amino)butyl)dithioalkyl)-2-(4-(bis(2-hydroxydodecyl)amino)butamido)-N-(3-(diethylamino)propyl)butamido) 4-((4-(bis(2-hydroxydecyl)amino)butyl)dithioalkyl)-2-(4-(bis(2-hydroxydodecyl)amino)butamido)-N-(2-(dimethylamino)ethyl)butamido) 4-((4-(bis(2-hydroxydecyl)amino)butyl)dithioalkyl)-2-(4-(bis(2-hydroxydodecyl)amino)butamido)-N-(2-(dimethylamino)ethyl)-N-methylbutamido) 4-((4-(bis(2-hydroxydecyl)amino)butyl)dithioalkyl)-2-(4-(bis(2-hydroxydodecyl)amino)butamido)-N-(2-(diethylamino)ethyl)-N-methylbutamido) 4-((4-(bis(2-hydroxydecyl)amino)butyl)dithioalkyl)-2-(4-(bis(2-hydroxydodecyl)amino)butamido)-N-(3-(2-methyl-1H-imidazol-1-yl)propyl)butamido 4-((4-(bis(2-hydroxydecyl)amino)butyl)dithioalkyl)-2-(4-(bis(2-hydroxydodecyl)amino)butamido)-N-(4-morpholinobutyl)butamido) 4-((4-(bis(2-hydroxydecyl)amino)butyl)dithioalkyl)-2-(4-(bis(2-hydroxydodecyl)amino)butamido)-N-(4-(pyrrolidone-1-yl)butyl)butamido) 2-(4-(bis(2-hydroxydodecyl)amino)butamido)-4-((4-(bis(2-hydroxydodecyl)amino)butyl)dithio)-N-(3-(dimethylamino)propyl)butamido) 2-(4-(bis(2-hydroxydodecyl)amino)butyramido)-4-((4-(bis(2-hydroxydodecyl)amino)butyl)dithio)-N-(2-(diethylamino)ethyl)butyramido) 2-(4-(bis(2-hydroxydodecyl)amino)butyramido)-4-((4-(bis(2-hydroxydodecyl)amino)butyl)dithioalkyl)-N-(3-(diethylamino)propyl)butyramido 2-(4-(bis(2-hydroxydodecyl)amino)butyramido)-4-((4-(bis(2-hydroxydodecyl)amino)butyl)dithio)-N-(2-(dimethylamino)ethyl)butyramido 2-(4-(bis(2-hydroxydodecyl)amino)butamido)-4-((4-(bis(2-hydroxydodecyl)amino)butyl)dithio)-N-(2-(dimethylamino)ethyl)-N-methylbutamido) 2-(4-(bis(2-hydroxydodecyl)amino)butamido)-4-((4-(bis(2-hydroxydodecyl)amino)butyl)dithio)-N-(2-(diethylamino)ethyl)-N-methylbutamido) 2-(4-(bis(2-hydroxydodecyl)amino)butamido)-4-((4-(bis(2-hydroxydodecyl)amino)butyl)dithio)-N-(3-(4-(2-hydroxyethyl)piperazin-1-yl)propyl)butamido) 2-(4-(bis(2-hydroxydodecyl)amino)butamido)-4-((4-(bis(2-hydroxydodecyl)amino)butyl)dithio)-N-(4-morpholinobutyl)butamido) 2-(4-(bis(2-hydroxydodecyl)amino)butamido)-4-((4-(bis(2-hydroxydodecyl)amino)butyl)dithioalkyl)-N-(4-(pyrrolidone-1-yl)butyl)butamido) And its pharmaceutically acceptable salts and esters.

10. A method for preparing a compound according to any one of the preceding claims, the method comprising the following steps: Compounds having formula (II): (II) Reaction with compounds having formula (III): (III) Where m, n, p, R 1 R 2 R 3 R 4 R 5 R 6 and R 7 As defined in any one of claims 1 to 8.

11. The method according to claim 10, further comprising preparing the compound having formula (II) by: making the compound having formula (IV) (IV) and Compounds having formula (V): (V) Reaction with compounds having formula (VI) (WE) Where m, n, and R 3 R 4 R 5 R 6 and R 7 As defined in any one of claims 1 to 6.

12. A method for preparing a compound according to any one of claims 1 to 9, the method comprising the following one-pot step: preparing a compound having formula (III): (III) Compounds having formula (IV) (IV) Compounds having formula (V): (V) Reaction with compounds having formula (VI): (WE) Where m, n, p, R 1 R 2 R 3 R 4 R 5 R 6 and R 7 As defined in any one of claims 1 to 8.

13. The preparation method according to claim 11 or claim 12, further comprising the step of preparing the compound having formula (IV) by: in the presence of a coupling agent, causing the compound having formula (VII) to: (VII) Where X represents an H or hydroxyl protecting group, and R6 and R7 are as defined in formula (I); Reaction with compounds having formula (VIII) (VIII) Where R 6 and R 7 As defined in any one of claims 1 to 6, and Hal represents a halogen atom. Optionally, the obtained product is then deprotected in an acidic medium, wherein X is a hydroxyl protecting group.

14. A lipid nanoparticle (LNP), such as a liposome, said LNP comprising a compound having formula (I) according to any one of claims 1 to 9, and optionally encapsulating a nucleic acid.

15. The LNP of claim 14, wherein the nucleic acid is mRNA encoding a peptide or protein.

16. The LNP according to claim 14 or 15, further comprising one or more components selected from: stealth lipids, structural lipids, and accessory lipids.

17. A pharmaceutical composition comprising LNP according to any one of claims 14 to 16, and one or more pharmaceutically acceptable excipients.

18. The LNP according to any one of claims 14 to 16, for delivering nucleic acids into a patient's cells.

19. The LNP according to any one of claims 14 to 16, used as a vaccine, such as a vaccine against influenza or a respiratory syncytial virus (RSV) vaccine.

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