Lipids, formulations and uses

Abstract

The present disclosure provides compounds, compositions, formulations, and methods for delivering active agents (e.g., nucleic acids). In particular, the present disclosure provides modified lipids (e.g., lipids conjugated with 1-methyl-D-tryptophan) for use in the delivery of active agents and compositions and vaccines thereof.

Description

[Technical field]

[0001] CROSS-REFERENCE TO RELATED APPLICATIONS This application claims the benefit of U.S. Provisional Application No. 63 / 334,437, filed April 25, 2022, the contents of which are incorporated herein by reference in their entirety.

[0002] The present disclosure provides compounds, compositions, formulations, and methods for delivering active agents (eg, nucleic acids). [Background technology]

[0003] The ability to deliver macromolecules and active drugs (e.g., nucleic acids) into cells is of great interest for therapy, diagnostics, reagents, and biological assays. Recently, the delivery of siRNA and mRNA-based therapeutics is currently in clinical use for coronavirus disease (COVID-19) and cancer. However, the delivery of nucleic acids is challenging due to instability, low permeability, and unwanted activation of the innate immune response. To facilitate the in vivo delivery of nucleic acids, a safe, effective, and stable delivery system is needed.

[0004] Lipid nanoparticles (LNPs) and cationic liposomes have already been demonstrated as effective delivery systems for nucleic acids. The composition of lipid nanoparticles or cationic liposomes includes cationic lipids or ionized lipids and other types of lipids, which are functionalized to encapsulate nucleic acids and induce efficient transfection in vivo. Although various such lipid nanoparticle compositions have been demonstrated, improvements in safety, efficacy, and specificity are still lacking. Summary of the Invention

[0005] In one embodiment, a compound of formula (I): [ka] or a pharma- ceutically acceptable salt thereof is disclosed herein. (In the formula, R 1 is hydrogen or C1-C6 alkyl; X is O, NR 2 , S, and a bond; R 2 is hydrogen, C1-C6 alkyl, C2-C6 alkenyl, C2-C6 alkynyl, C3-C6 cycloalkyl, or halo-C1-C6-alkyl; L is C8-C 80 Alkyl, C8-C 80 Alkenyl, C8-C 80 Alkynyl, C8-C 80 Heteroalkyl, C8-C 80 Heteroalkenyl, and C8-C 80 heteroalkynyl, each of which is optionally substituted with one or more substituents selected from hydroxy and amino.

[0006] In some embodiments, R 1 is methyl.

[0007] In some embodiments, L is C 12 -C 40 Alkyl and C 12 -C 40 In some embodiments, L is selected from the group consisting of aryl, aryl, and alkenyl. In some embodiments, L has the formula (A): [ka] (In the formula, n is 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, or 40; R a and R b are each independently C6-C 40 Alkyl, C6-C 40 Alkenyl, C6-C 40 Heteroalkyl and C6-C 40 heteroalkenyl). In some embodiments, n is 1, 2, 3, 4, 5, 6, 7, or 8.

[0008] In some embodiments, L has the formula (D), (E), or (F): [ka] (In the formula, n, p, and q are each independently 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, or 40; R a1 and R a2 are each independently C6-C 40 Alkyl and C6-C 40 alkenyl). In some embodiments, n, p, and q are each independently 1, 2, 3, 4, 5, 6, 7, or 8.

[0009] In some embodiments, L is selected from: [ka] [ka] [ka]

[0010] In some embodiments, the compound is selected from the group consisting of: [ka]

[0011] In another aspect, a lipid composition is disclosed that comprises a compound disclosed herein (e.g., a compound of Formula (I)) or a pharma- ceutically acceptable salt thereof. In some embodiments, the lipid composition further comprises one or more of a cationic lipid and / or an ionizable lipid, a phospholipid, a neutral or non-cationic lipid, or a combination thereof.

[0012] In some embodiments, the lipid composition further comprises a structured lipid, hi some embodiments, the structured lipid is selected from the group consisting of cholesterol, fecosterol, sitosterol, ergosterol, campesterol, stigmasterol, brassicasterol, tomatidine, cholesteryl hemisuccinate, cholesteryl sulfate, ursolic acid, alpha-tocopherol, and mixtures thereof.

[0013] In some embodiments, the lipid composition further comprises a polyethylene glycol (PEG)-lipid conjugate.

[0014] In some embodiments, the lipid composition further comprises at least one active agent. In some embodiments, the at least one active agent is encapsulated in a liposome, lipid nanoparticle, or micelle in the lipid composition. In some embodiments, the at least one active agent comprises a nucleic acid or a protein. In some embodiments, the nucleic acid encodes an antigen or a functional fragment thereof.

[0015] In some embodiments, the at least one active agent comprises an RNA. In some embodiments, the RNA is selected from the group consisting of small interfering RNA (siRNA), asymmetric interfering RNA (aiRNA), microRNA (miRNA), dicer substrate RNA (dsRNA), small hairpin RNA (shRNA), messenger RNA (mRNA), ribozyme, and mixtures thereof.

[0016] In some embodiments, the at least one active agent comprises an immunomodulatory agent, a chemotherapeutic agent, a steroid, an analgesic agent, an antimicrobial agent, or a combination thereof.

[0017] In another aspect, disclosed herein is a pharmaceutical composition comprising an effective amount of a compound disclosed herein (e.g., a compound of Formula (I)) or a pharma- ceutically acceptable salt thereof, or a lipid composition disclosed herein, and a pharma- ceutically acceptable carrier.

[0018] In another aspect, disclosed herein is a vaccine comprising an effective amount of a compound disclosed herein (e.g., a compound of formula (I), or a pharma- ceutically acceptable salt thereof), a lipid composition described herein, or a pharmaceutical composition disclosed herein (e.g., a pharmaceutical composition comprising a compound of formula (I), or a pharma- ceutically acceptable salt thereof, or a lipid composition described herein); and an antigen or a nucleic acid encoding same.

[0019] In some embodiments, the antigen is a tumor antigen, an autoantigen, or an antigen from an infectious disease. In some embodiments, the nucleic acid is messenger RNA (mRNA).

[0020] In another aspect, disclosed herein is a method for delivering an active agent to a cell, the method comprising contacting the cell with a lipid composition disclosed herein, or a pharmaceutical composition disclosed herein (e.g., a pharmaceutical composition comprising a compound of Formula (I), or a pharma- ceutical acceptable salt thereof, or a lipid composition described herein), or a vaccine disclosed herein (e.g., a vaccine comprising a compound of Formula (I), or a pharma- ceutical acceptable salt thereof, or a lipid composition described herein) in a subject in need thereof.

[0021] In another aspect, disclosed herein is a method for delivering an active agent to a subject, the method comprising administering to a subject in need thereof a lipid composition disclosed herein, or a pharmaceutical composition disclosed herein (e.g., a pharmaceutical composition comprising a compound of formula (I), or a pharma- ceutical acceptable salt thereof, or a lipid composition described herein), or a vaccine disclosed herein (e.g., a vaccine comprising a compound of formula (I), or a pharma- ceutical acceptable salt thereof, or a lipid composition described herein).

[0022] In another aspect, disclosed herein is a method for producing a polypeptide of interest in a cell, the method comprising contacting the cell with a lipid composition disclosed herein, or a pharmaceutical composition disclosed herein (e.g., a pharmaceutical composition comprising a compound of Formula (I) or a pharma- ceutically acceptable salt thereof, or a lipid composition described herein), or a vaccine disclosed herein (e.g., a vaccine comprising a compound of Formula (I) or a pharma- ceutically acceptable salt thereof, or a lipid composition described herein) in a subject in need thereof, wherein the lipid composition or pharmaceutical composition comprises an mRNA encoding the polypeptide of interest. In some embodiments, the cell is present in a subject, and the contacting comprises administration to the subject.

[0023] In another aspect, a method of treating or preventing a disease or disorder is disclosed herein, the method comprising administering to a subject in need thereof an effective amount of a compound disclosed herein (e.g., a compound of formula (I), or a pharma- ceutically acceptable salt thereof), or a lipid composition disclosed herein, or a pharmaceutical composition disclosed herein (e.g., a pharmaceutical composition comprising a compound of formula (I), or a pharma- ceutically acceptable salt thereof, or a lipid composition described herein), or a vaccine disclosed herein (e.g., a vaccine comprising a compound of formula (I), or a pharma- ceutically acceptable salt thereof, or a lipid composition described herein).

[0024] In some embodiments, the disease or disorder comprises cancer, an autoimmune disease, an inflammatory disease, or an infectious disease. In some embodiments, the disease or disorder is cancer. In some embodiments, the subject has cancer, has had cancer, is susceptible to cancer, or has a family history of cancer. In some embodiments, the cancer comprises a solid tumor or a hematological cancer. In some embodiments, the cancer is a metastatic cancer. In some embodiments, the method inhibits or eliminates metastasis of the cancer, reduces tumor growth, prevents tumor recurrence, or any combination thereof. In some embodiments, the administration comprises an initial immunization and at least one subsequent immunization.

[0025] In some embodiments, the subject is a human.

[0026] In some embodiments, the method further comprises administering at least one additional active agent, in some embodiments, the at least one additional active agent comprises an immunomodulatory agent, a chemotherapeutic agent, a nucleic acid, a steroid, an analgesic agent, an antimicrobial agent, or a combination thereof.

[0027] Other aspects and embodiments of the present disclosure will become apparent in light of the following detailed description. DETAILED DESCRIPTION OF THE PREFERRED EMBODIMENTS

[0028] Described herein are modified lipids for use in liposomal and lipid nanoparticle delivery of active agents (e.g., miRNA) that facilitate efficient encapsulation, improved delivery, and enhanced downstream efficacy (e.g., by macrophage polarization). The modified lipids include cationic or ionizable lipids conjugated to amino acids or IDO inhibitors (e.g., 1-methyl-D-tryptophan).

[0029] The section headings used in this section and throughout this disclosure are for organizational purposes only and are not intended to be limiting.

[0030] 1.Definition As used herein, the terms "comprise," "include," "having," "has," "can," "contain," and variations thereof are intended to be open-ended transitional phrases, terms, or words that do not exclude the possibility of additional acts or structures. The singular forms "a," "and," and "the" also include plural referents unless the context clearly indicates otherwise. The present disclosure also contemplates other embodiments that "comprising," "consisting of," and "consisting essentially of" the embodiments or elements presented herein, whether or not expressly stated.

[0031] With respect to references to numerical ranges herein, each intervening numerical value therebetween is expressly contemplated with the same degree of precision. For example, for the range 6 to 9, the numbers 7 and 8 are contemplated in addition to 6 and 9, and for the range 6.0 to 7.0, the numbers 6.0, 6.1, 6.2, 6.3, 6.4, 6.5, 6.6, 6.7, 6.8, 6.9, and 7.0 are expressly contemplated.

[0032] Unless otherwise defined herein, scientific and technical terms used in connection with this disclosure shall have the meanings commonly understood by those skilled in the art. The meaning and scope of the terms must be clear, but in the event of any potential ambiguity, the definitions provided herein shall take precedence over any dictionary or external definitions. Furthermore, unless otherwise required by context, singular terms shall include the plural, and plural terms shall include the singular. All publications, patent applications, patents, and other references mentioned herein are incorporated by reference in their entirety.

[0033] The term "contact" as used herein means to contact or to be in contact with, to be in contact with, or to come into contact with. The term "contact" as used herein refers to the state or condition of contact or direct or local proximity.

[0034] As used herein, "polynucleotide", "oligonucleotide" or "nucleic acid" refers to at least two nucleotides covalently linked together. A polynucleotide may be DNA, both genomic and cDNA, RNA, or hybrid, and a polynucleotide may contain a combination of deoxyribonucleotides and ribonucleotides, and a combination of bases (e.g., uracil, adenine, thymine, cytosine, guanine, inosine, xanthine, hypoxanthine, isocytosine, and isoguanine). A nucleic acid may contain non-natural nucleotides, modified nucleotides, and / or non-nucleotide building blocks (e.g., "nucleotide analogs") that allow DNA or RNA to exhibit the same function as natural nucleotides. A nucleic acid may be obtained by chemical synthesis or recombinant methods. A polynucleotide may be single-stranded or double-stranded, or may contain portions of both double-stranded and single-stranded sequences. The depiction of a single strand also defines the sequence of the complementary strand. Thus, a nucleic acid also contains the complementary strand of a depicted single strand. Many variants of a nucleic acid can be used for the same purpose as a given nucleic acid. Thus, a nucleic acid also encompasses substantially identical nucleic acids and their complements.

[0035] A "peptide" or "polypeptide" is a linked sequence of two or more amino acids joined by peptide bonds. Peptides and polypeptides include proteins such as binding proteins, receptors, and antibodies. The terms "polypeptide" and "protein" are used interchangeably herein.

[0036] As used herein, "nucleic acid" or "nucleic acid sequence" refers to a polymer or oligomer of pyrimidine and / or purine bases, preferably cytosine, thymine, and uracil, and adenine and guanine, respectively (see Albert L. Lehninger, Principles of Biochemistry, at 793-800 (Worth Pub. 1982)). The present technology contemplates any deoxyribonucleotide, ribonucleotide, or peptide nucleic acid component, and any chemical variants thereof, such as methylated, hydroxymethylated, or glycosylated forms of these bases. The polymer or oligomer may be heterogeneous or homogeneous in composition, and may be isolated from naturally occurring sources or may be artificially or synthetically produced. Furthermore, the nucleic acid may be DNA or RNA, or a mixture thereof, and may exist permanently or transiently in single-stranded or double-stranded form, including homoduplexes, heteroduplexes, and hybrid states. In some embodiments, the nucleic acid or nucleic acid sequence comprises other types of nucleic acid structures, such as, for example, a DNA / RNA helix, a peptide nucleic acid (PNA), a morpholino nucleic acid (see, e.g., Braaschand Corey, Biochemistry, 41(14):4503-4510 (2002), and U.S. Pat. No. 5,034,506), a locked nucleic acid (LNA; see, Wahlestedtetal., Proc. Natl. Acad. Sci. USA, 97:5633-5638 (2000)), a cyclohexenyl nucleic acid (see, Wang, J. Am. Chem. Soc., 122:8595-8602 (2000)), and / or a ribozyme.Thus, the term "nucleic acid" or "nucleic acid sequence" may also encompass a strand that includes non-natural nucleotides, modified nucleotides, and / or non-nucleotide building blocks (e.g., "nucleotide analogs") that may perform the same function as natural nucleotides; furthermore, the term "nucleic acid sequence" as used herein refers to an oligonucleotide, nucleotide, or polynucleotide, and fragments or portions thereof, as well as DNA or RNA of genomic or synthetic origin, which may be single-stranded or double-stranded and may represent the sense or antisense strand. The terms "nucleic acid," "polynucleotide," "nucleotide sequence," and "oligonucleotide" are used interchangeably. They refer to polymeric forms of nucleotides of any length, either deoxyribonucleotides or ribonucleotides, or analogs thereof.

[0037] As used herein, the terms "providing," "administering," and "introducing" are used interchangeably herein and refer to the placement of an active agent in a subject by a method or route that results in at least partial localization to a desired site.

[0038] A "subject" or "patient" may be human or non-human, and may include animal strains or species used as "model systems" for research purposes, such as the mouse model described herein. Similarly, a patient may include an adult or a minor (e.g., a child). Furthermore, a patient may refer to any organism, preferably a mammal (e.g., human and non-human), that may benefit from administration of the compositions contemplated herein. Examples of mammals include, but are not limited to, any member of the class Mammalia; humans, non-human primates (e.g., chimpanzees and other ape and monkey species); livestock animals (e.g., cows, horses, sheep, goats, wild boars); farm animals (e.g., rabbits, dogs, and cats); laboratory animals, including rodents (e.g., rats, mice, and guinea pigs, etc.). Examples of non-mammals include, but are not limited to, birds, fish, and the like. In one embodiment, the mammal is a human.

[0039] As used herein, "treat", "treating" and the like refer to slowing, stopping, or reversing the progression of a disease or disorder when a compound or composition described herein is provided to a suitable control subject. The term also refers to reversing the progression of such a disease or disorder to the point where symptoms are eliminated or significantly reduced. Thus, "treating" refers to applying or administering a composition described herein to a subject, the subject having a disease or symptoms of a disease, with the purpose of curing, curing, alleviating, mitigating, altering, treating, improving, enhancing, or affecting the disease or symptoms of the disease.

[0040] Definitions of certain functional groups and chemical terms are described in more detail below. For purposes of this disclosure, chemical elements are defined as defined in the Periodic Table of the Elements (CAS version, Handbook of Chemistry and Physics, 75 th In addition, general principles of organic chemistry, as well as specific functional moieties and reactivities, are described in: Sorrell, Organic Chemistry, 2001, pp. 111-115, 1997; and in: Sorrell, Organic Chemistry, 2001, pp. 111-115, 1997. nd edition,University Science Books,Sausalito,2006;Smith,March's Advanced Organic Chemistry:Reactions,Mechanism,and Structure,7 th Edition,John Wiley & Sons,Inc.,New York,2013;Larock,Comprehensive Organic Transformations,3 rd Edition, John Wiley & Sons, Inc., New York, 2018; and Carruthers, Some Modern Methods of Organic Synthesis, 3 rdEdition, Cambridge University Press, Cambridge, 1987 (the entire contents of each of which are incorporated herein by reference).

[0041] The term "alkyl" as used herein refers to a saturated straight or branched hydrocarbon chain. Representative examples of alkyl include, but are not limited to, methyl, ethyl, n-propyl, isopropyl, n-butyl, sec-butyl, isobutyl, tert-butyl, n-pentyl, isopentyl, neopentyl, n-hexyl, 3-methylhexyl, 2,2-dimethylpentyl, 2,3-dimethylpentyl, 4,4-dimethylpentan-2-yl, n-heptyl, n-octyl, n-nonyl, n-decyl, undecyl, dodecyl, tridecyl, tetradecyl, pentadecyl, hexadecyl, heptadecyl, octadecyl, nonadecyl, and icosyl.

[0042] The term "alkenyl" as used herein refers to a straight or branched hydrocarbon chain containing at least one carbon-carbon double bond. The double bond(s) can be located at any position of the hydrocarbon chain. Representative examples of alkenyl include, but are not limited to, ethenyl, 2-propenyl, 2-methyl-2-propenyl, 3-butenyl, 4-pentenyl, 5-hexenyl, 2-heptenyl, 2-methyl-1-heptenyl, and 3-decenyl.

[0043] The term "alkynyl" as used herein refers to a straight or branched hydrocarbon chain containing at least one carbon-carbon triple bond. The triple bond(s) may be located at any position of the hydrocarbon chain. Representative examples of alkynyl include, but are not limited to, ethynyl, propynyl, and butynyl.

[0044] The term "amino" as used herein refers to the group -NH2. The term "alkylamino" as used herein refers to the group -NHR, where R is an alkyl group as defined herein. The term "dialkylamino" as used herein refers to the group -NR2, where each R is independently an alkyl group as defined herein.

[0045] The term "cycloalkyl" as used herein refers to a saturated carbocyclic ring system containing 3 to 10 carbon atoms and 0 heteroatoms. Cycloalkyls can be monocyclic, bicyclic, bridged, fused, or spirocyclic. Representative examples of cycloalkyls include, but are not limited to, cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, cycloheptyl, cyclooctyl, cyclononyl, cyclodecyl, adamantyl, bicyclo[2.2.1]heptanyl, bicyclo[3.2.1]octanyl, and bicyclo[5.2.0]nonanyl.

[0046] The term "halogen" or "halo" as used herein means F, Cl, Br, or I.

[0047] The term "haloalkyl" as used herein refers to an alkyl group, as defined herein, in which at least one hydrogen atom (e.g., 1, 2, 3, 4, 5, 6, 7, or 8 hydrogen atoms) is replaced with a halogen. In some embodiments, each hydrogen atom of the alkyl group is replaced with a halogen. Representative examples of haloalkyl include, but are not limited to, fluoromethyl, difluoromethyl, trifluoromethyl, 2-fluoroethyl, 2,2,2-trifluoroethyl, and 3,3,3-trifluoropropyl.

[0048] As used herein, the term "heteroalkyl" refers to an alkyl group in which one or more of the carbon atoms (and associated hydrogen atoms) are each independently selected from the group consisting of a heteroatom group (e.g., -NH-, -O-, -S-, -S(O)-, -S(O)2-, -OP(O)(O -)O-, etc. As an example, 1, 2, 3, 4, 5, 6, or more carbon atoms may be independently replaced with the same or different heteroatom groups. Heteroalkyl groups can also contain one or more carbonyl moieties (i.e., a carbon atom of the alkyl group is oxidized to a -C(O)- group).

[0049] The term "heteroalkenyl" as used herein refers to an alkenyl group, as defined herein, in which one or more of the carbon atoms (and any associated hydrogen atoms) are each independently replaced with a heteroatom group, such as -NH-, -O-, -S-, -S(O)-, -S(O)2-. As an example, 1, 2, 3, 4, 5, 6, or more carbon atoms may be independently replaced with the same or different heteroatom groups. Heteroalkenyl groups may also contain one or more carbonyl moieties (i.e., a carbon atom of an alkyl group is oxidized to a -C(O)- group).

[0050] The term "heteroalkynyl" as used herein refers to an alkynyl group, as defined herein, in which one or more of the carbon atoms (and any associated hydrogen atoms) are each independently replaced with a heteroatom group, such as -NH-, -O-, -S-, -S(O)-, -S(O)2-. As an example, 1, 2, 3, 4, 5, 6, or more carbon atoms may be independently replaced with the same or different heteroatom groups. Heteroalkynyl groups may also contain one or more carbonyl moieties (i.e., a carbon atom of an alkyl group is oxidized to a -C(O)- group).

[0051] As used herein, the term "hydroxy" refers to an --OH group.

[0052] As used herein, the term "substituent" refers to a group substituted on an atom of a designated group.

[0053] Where a group or moiety can be substituted, the term "substituted" indicates that one or more (e.g., 1, 2, 3, 4, 5, or 6, in some embodiments, 1, 2, or 3, and in other embodiments, 1 or 2) hydrogen atoms on the group designated by the phrase can be replaced with the specified group recited or with suitable substituents known to those of skill in the art (e.g., one or more of the groups recited below), provided that the normal valence of the given atom is not exceeded. Substituents include, but are not limited to, alkyl, alkenyl, alkynyl, alkoxy, acyl, amino, amido, amidino, aryl, azido, carbamoyl, carboxyl, carboxyl ester, cyano, cycloalkyl, cycloalkenyl, guanidino, halo, haloalkyl, haloalkoxy, heteroalkyl, heteroaryl, heterocyclyl, hydroxy, hydrazino, imino, oxo, nitro, phosphate, phosphonate, sulfonic acid, sulfonamide, thiol, thione, thioxo, or combinations thereof.

[0054] As used herein, in chemical structures, the indications: [ka] represents the point of attachment of one moiety to another.

[0055] In some cases, the number of carbon atoms in a hydrocarbyl substituent (e.g., alkylalkenyl) is indicated by the prefix “C x -C y " (where x is the minimum number of carbon atoms in the substituent and y is the maximum number of carbon atoms in the substituent). Thus, for example, "C1-C3 alkyl" refers to an alkyl substituent containing 1 to 3 carbon atoms.

[0056] In the compounds described herein, groups and substituents thereof may be selected according to the allowed valences of atoms and substituents such that the selection and substitution result in stable compounds that do not spontaneously undergo transformation, e.g., by rearrangement, cyclization, elimination, and the like.

[0057] Where substituents are designated in a conventional chemical formula written from left to right, they optionally include the substituents resulting from writing the structure from right to left. For example, -CHO- is intended to include -OCH-, -C(O)NH- is intended to include -NHC(O)-.

[0058] Preferred methods and materials are described below, although methods and materials similar or equivalent to those described herein can be used in the practice or testing of this disclosure. All publications, patent applications, patents, and other references mentioned herein are incorporated by reference in their entirety. The materials, methods, and examples disclosed herein are illustrative only and are not intended to be limiting.

[0059] 2.Compound In one embodiment, a compound of formula (I): [ka] or a pharma- ceutically acceptable salt thereof, wherein: R 1 is hydrogen or C1-C6 alkyl; X is O, NR 2 , S, and a bond; R 2 is hydrogen, C1-C6 alkyl, C2-C6 alkenyl, C2-C6 alkynyl, C3-C6 cycloalkyl, or haloC1-C6 alkyl; L is C8-C 80 Alkyl, C8-C 80 Alkenyl, C8-C 80 Alkynyl, C8-C 80 Heteroalkyl, C8-C 80 Heteroalkenyl, and C8-C 80 heteroalkynyl, each of which is optionally substituted with one or more substituents selected from hydroxy and amino.

[0060] In some embodiments, R 1 is hydrogen. In some embodiments, R1 is C-C alkyl, and in some embodiments, R 1 is methyl.

[0061] In some embodiments, X is O or a bond. In some embodiments, X is O. In some embodiments, X is a bond. In some embodiments, X is NR 2 and R 2 is selected from hydrogen and C1-C6 alkyl. In some embodiments, X is NH. In some embodiments, X is NR 2 and R 2 In some embodiments, X is S.

[0062] In some embodiments, L is a lipid moiety having at least 8 carbon atoms, and the lipid moiety can be derived from any suitable lipid (e.g., fatty alcohol, fatty acid, phospholipid, steroid, or synthetic lipid). In some embodiments, the lipid moiety is derived from a lipid having a functional group (e.g., a hydroxyl group, a carboxylic acid group, or an amino group), and the lipid moiety is attached to the compound of formula (I) via the functional group. For example, in some embodiments, L is a lipid moiety having at least 8 carbon atoms, and in such embodiments, the group X in formula (I) is derived from a functional group; for example, when the lipid moiety is derived from an aliphatic alcohol, X is O.

[0063] In some embodiments, L is C-C 80 Alkyl, C8-C 80 Alkenyl, C8-C 80 Alkynyl, C8-C 80 Heteroalkyl, C8-C 80 Heteroalkenyl, and C8-C 80 heteroalkynyl, each of which is optionally substituted with one or more substituents selected from hydroxy and amino.

[0064] For example, in some embodiments, L is C-C 80Alkyl and C8-C 80 In some embodiments, L is selected from C-C alkenyl. 40 Alkyl and C8-C 40 In some embodiments, L is selected from C 12 -C 40 Alkyl and C 12 -C 40In some such embodiments, L is derived from a saturated or unsaturated fatty alcohol. In some embodiments, L is derived from: linoleyl alcohol ((9Z,12Z)-octadeca-9,12-dien-1-ol), myristyl alcohol (1-tetradecanol), palmitoleyl alcohol ((Z)-hexadec-9-en-1-ol), oleyl alcohol ((Z)-octadec-9-en-1-ol), elaidyl alcohol (trans-9-octadecenol), cis-butyl alcohol (cis-butyl alcohol ... Xenyl alcohol (cis-11-octadecenol), gadoleyl alcohol ((Z)-icos-9-en-1-ol), 11-eicosenol, erucyl alcohol (cis-13-docosenol), 15-tetracosen-1-ol, eicosadienyl alcohol (icosa-11,14-dien-1-ol), linolenyl alcohol ((9Z,12Z,15Z)-9,12,15-octadecatrien-1-ol), ol), γ-linolenyl alcohol ((6E,9E,12E)-octadeca-6,9,12-trien-1-ol), eleostearyl alcohol (octadeca-9,11,13-trien-1-ol), icos-5,8,11-trien-1-ol, icos-13-en-1-ol, icos-11,14-17-trien-1-ol, octadeca-6,9,12,15-tetraen-1-ol, arachidonic acid 4E,6Z-Hexadecadien-1-ol, icosa-5,8,11,14,17-pentaen-1-ol, docosahexaenoyl alcohol (docosa-4,7,10,13,16,19-hexaen-1-ol), docosa-7,10,13,16,19-pentaen-1-ol, tetracosa-6,9,12,15,18,21-Hexaen-1-ol, Capryl Alcohol (1-Octanol), Pelargonic Alcohol (1-Nonanol), Decyl Alcohol (1-Decanol), Undecyl Alcohol (1-Undecanol), Lauryl Alcohol (1-Dodecanol), Tridecyl Alcohol (1-Tridecanol), Myristyl Alcohol (1-Tetradecanol), Pentadecyl Alcohol (1-Pentadecanol), Cetyl Alcohol (1-Hexadecanol), Palmitoleic Alcohol (cis-9-Hexadecen-1-ol), Heptadecyl Alcohol (1-n-Heptadecanol), Stearyl Alcohol (1-Octadecanol), Oleyl Alcohol (1-Octadecenol), Nonadecanyl Alcohol (1-Nonadecanol), Arachidyl Alcohol (1-Eicosanol), Heneicosyl Alcohol 1-Heneicosanol, Behenyl Alcohol, 1-Docosanol, Erucyl Alcohol, 1-Tricosyl Alcohol, 1-Tetracosanol, Pentacosyl Alcohol, 1-Pentacosanol, Seryl Alcohol, 1-Heptacosanol, Montanyl Alcohol, 1-Octacosanol, 1-Nonacosanol, Myricyl Alcohol, 1-Triacontanol, 1-Hentriacontanol, 1-Dotriacontanol, Laceryl Alcohol, 1-Tritriacontanol, Gedil Alcohol, 1-Tetratriacontanol, 1-Hexatriacontanol, 1-Heptatriacontanol, 1-Octatriacontanol, Nonatriacontan-1-ol, or 1-Tetracontanol.

[0065] In some embodiments, L is C-C 80 Heteroalkyl, C8-C 80 Heteroalkenyl, and C8-C 80 In such embodiments, L may be derived from a lipid that includes one or more heteroatom groups (e.g., -O-, -NH-, -C(O)-, etc., or combinations thereof (e.g., -C(O)O- groups).

[0066] For example, in some embodiments, L has the formula (A): [ka] (In the formula, n is 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, or 40; R a and R b are each independently C6-C 40 Alkyl, C6-C 40 Alkenyl, C6-C 40 Heteroalkyl and C6-C 40 heteroalkenyl).

[0067] For example, in some embodiments, n is 1, 2, 3, 4, 5, 6, 7, or 8. In some embodiments, n is 2, 3, or 4. In some embodiments, n is 2. In some embodiments, n is 3. In some embodiments, n is 4.

[0068] In some embodiments, R a and R b are each independently C6-C 40 Alkyl and C6-C 40 For example, in some embodiments, R a and R bare each independently selected from the following: n-hexyl, n-heptyl, n-octyl, n-nonyl, n-decyl, undecyl, dodecyl, tridecyl, tetradecyl, pentadecyl, hexadecyl, heptadecyl, octadecyl, nonadecyl, icosyl, henicosyl, docosyl, tricosyl, tetracosyl, pentacosyl, hexacosyl, heptacosyl, octacosyl, nonacosyl, triacontyl, hentriacontyl, dotriacontyl, tritriacontyl, tetratriacontyl, pentatriacontyl, pentyl, hexatriacontyl, heptatriacontyl, octatriacontyl, nonatriacontyl, tetracontyl, linoleyl ((9Z,12Z)-octadeca-9,12-dien-1-yl), palmitoleyl ((Z)-hexadec-9-en-1-yl), oleyl ((Z)-octadec-9-en-1-yl), elaidyl (trans-9-octadecenyl), cis-vaccenyl (cis-11-octadecenyl), gadoleyl ((Z)-icos-9-en-1-yl), 11-eicosenyl, elci cis-13-docosenyl, 15-tetracosen-1-yl, eicosadienyl (icosa-11,14-dien-1-yl), linolenyl ((9Z,12Z,15Z)-9,12,15-octadecatrien-1-yl), γ-linolenyl ((6E,9E,12E)-octadeca-6,9,12-trien-1-yl), eleostearyl (octadeca-9,11,13-trien-1-yl), icos-5,8,11-trien-1-yl, eicos-13-en-1-yl, icos-11,14-17- In some embodiments, R is selected from the group consisting of icosatetraen-1-yl, octadeca-6,9,12,15-tetraen-1-yl, arachidonyl ((5Z,8Z,11Z,14Z)-icosatetraen-1-yl), 4E,6Z-hexadecadien-1-yl, icosa-5,8,11,14,17-pentaen-1-yl, docosahexaenoyl (docosa-4,7,10,13,16,19-hexaen-1-yl), docosa-7,10,13,16,19-pentaen-1-yl, and tetracosa-6,9,12,15,18,21-hexaen-1-yl. a and R b are linoleyl, respectively.

[0069] In some embodiments, L has the formula (B) or (C): [ka] (In the formula, n and p are each independently 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, or 40; R a1 is C6-C 40 Alkyl and C6-C 40 alkenyl, R b is C6-C 40 Alkyl, C6-C 40 Alkenyl, C6-C 40 Heteroalkyl and C6-C 40 heteroalkenyl).

[0070] In some embodiments, n is 1, 2, 3, 4, 5, 6, 7, or 8. In some embodiments, n is 2, 3, or 4. In some embodiments, n is 2. In some embodiments, n is 3. In some embodiments, n is 4.

[0071] In some embodiments, p is 1, 2, 3, 4, 5, 6, 7, or 8. In some embodiments, p is 3, 4, 5, 6, or 7. In some embodiments, p is 3. In some embodiments, p is 4. In some embodiments, p is 5. In some embodiments, p is 6. In some embodiments, p is 7.

[0072] In some embodiments, R a1 is C6-C 24 Alkyl or C6-C 24 In some embodiments, R is alkenyl. a1is C9-C 22 Alkyl and C9-C 22 In some embodiments, R is selected from the group consisting of alkenyl and aryl. a1 are linear or branched C6, C7, C8, C9, C 10 , C 11 , C 12 , C 13 , C 14 , C 15 , C 16 , C 17 , C 18 , C 19 , C 20 , C 21 , C 22 , C 23 , or C 24 In some embodiments, R a1 are linear or branched C6, C7, C8, C9, C 10 , C 11 , C 12 , C 13 , C 14 , C 15 , C 16 , C 17 , C 18 , C 19 , C 20 , C 21 , C 22 , C 23 , or C 24 In some embodiments, R is selected from the group consisting of alkenyl and aryl. a1 is selected from linoleyl, n-nonyl, n-undecyl, henicosan-11-yl, pentadecan-7-yl, and heptadecan-9-yl.

[0073] In some embodiments, R b is C6-C 40 Alkyl and C6-C 40 For example, in some embodiments, R bis selected from the following: n-hexyl, n-heptyl, n-octyl, n-nonyl, n-decyl, undecyl, dodecyl, tridecyl, tetradecyl, pentadecyl, hexadecyl, heptadecyl, octadecyl, nonadecyl, icosyl, henicosyl, docosyl, tricosyl, tetracosyl, pentacosyl, hexacosyl, heptacosyl, octacosyl, nonacosyl, triacontyl, hentriacontyl, dotriacontyl, tritriacontyl, tetratriacontyl, pentatriacontyl, hexatriacontyl, Satriacontyl, heptatriacontyl, octatriacontyl, nonatriacontyl, tetracontyl, linoleyl ((9Z,12Z)-octadeca-9,12-dien-1-yl), palmitoleyl ((Z)-hexadec-9-en-1-yl), oleyl ((Z)-octadec-9-en-1-yl), elaidyl (trans-9-octadecenyl), cis-vaccenyl (cis-11-octadecenyl), gadoleyl ((Z)-icos-9-en-1-yl), 11-eicosenyl, erucyl (cis -13-docosenyl), 15-tetracosen-1-yl, eicosadienyl (icosa-11,14-dien-1-yl), linolenyl ((9Z,12Z,15Z)-9,12,15-octadecatrien-1-yl), γ-linolenyl ((6E,9E,12E)-octadeca-6,9,12-trien-1-yl), eleostearyl (octadeca-9,11,13-trien-1-yl), icos-5,8,11-trien-1-yl, eicos-13-en-1-yl, icos-11,14-17-trien-1-yl In some embodiments, R is selected from the group consisting of octadeca-6,9,12,15-tetraen-1-yl, arachidonyl ((5Z,8Z,11Z,14Z)-icosatetraen-1-yl), 4E,6Z-hexadecadien-1-yl, icosa-5,8,11,14,17-pentaen-1-yl, docosahexaenoyl (docosa-4,7,10,13,16,19-hexaen-1-yl), docosa-7,10,13,16,19-pentaen-1-yl, and tetracosa-6,9,12,15,18,21-hexaen-1-yl. b is linoleyl.

[0074] In some embodiments, L has the formula (D), (E), or (F): [ka] (In the formula, n, p, and q are each independently 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, or 40; R a1 and R a2 are each independently C6-C 40 Alkyl and C6-C 40 alkenyl).

[0075] In some embodiments, L has formula (D). In some embodiments, L has formula (E). In some embodiments, L has formula (F). In some embodiments, n is 1, 2, 3, 4, 5, 6, 7, or 8. In some embodiments, n is 2, 3, or 4. In some embodiments, n is 2. In some embodiments, n is 3. In some embodiments, n is 4.

[0076] In some embodiments, p and q are each independently selected from 1, 2, 3, 4, 5, 6, 7, or 8. In some embodiments, p and q are each independently selected from 3, 4, 5, 6, and 7. In some embodiments, p and q are each independently selected from 3, 5, and 7. In some embodiments, p and q are each 5. In some embodiments, p and q are each 6. In some embodiments, p and q are each 7.

[0077] In some embodiments, R a1 and R a2 are each independently C6-C 24 Alkyl and C6-C 24In some embodiments, R is selected from the group consisting of alkenyl and aryl. a1 and R a2 are each independently C9-C 22 Alkyl and C9-C 22 In some embodiments, R is selected from the group consisting of alkenyl and aryl. a1 and R a2 are each independently a linear or branched C6, C7, C8, C9, C 10 , C 11 , C 12 , C 13 , C 14 , C 15 , C 16 , C 17 , C 18 , C 19 , C 20 , C 21 , C 22 , C 23 , or C 24 In some embodiments, R a1 and R a2 are each independently a linear or branched C6, C7, C8, C9, C 10 , C 11 , C 12 , C 13 , C 14 , C 15 , C 16 , C 17 , C 18 , C 19 , C 20 , C 21 , C 22 , C 23 , or C 24 In some embodiments, R is selected from the group consisting of alkenyl and aryl. a1 and R a2 are each independently selected from linoleyl, n-nonyl, n-undecyl, henicosan-11-yl, pentadecan-7-yl, and heptadecan-9-yl.

[0078] In some embodiments, L is derived from a steroid, for example, in some embodiments, L is derived from cholesterol, beta-sisterol, or BHEM cholesterol.

[0079] In some embodiments, L is selected from: [ka] [ka] [ka]

[0080] Compounds may exist as stereoisomers in which asymmetric or chiral centers exist. Stereoisomers are "R" or "S" depending on the configuration of the substituents around the chiral carbon atom. The terms "R" and "S" as used herein are configurations as defined in: IUPAC 1974 Recommendations for Section E, Fundamental Stereochemistry, in Pure Appl. Chem., 1976, 45:13-30. The present disclosure contemplates various stereoisomers and mixtures thereof, which are specifically included within the scope of the present disclosure. Stereoisomers include enantiomers and diastereomers, as well as mixtures of enantiomers or diastereomers. Individual stereoisomers of the compounds may be prepared synthetically from commercially available starting materials containing asymmetric or chiral centers, or by preparing racemic mixtures followed by resolution methods well known to those skilled in the art. These resolution techniques are exemplified by: (1) binding the mixture of enantiomers to a chiral auxiliary, separating the resulting mixture of diastereomers by recrystallization or chromatography, and, optionally, isolating the optically pure products from the auxiliary as described in Furniss, Hannaford, Smith, and Tatchell, “Vogel's Textbook of Practical Organic Chemistry,” 5th edition (1989), Longman Scientific & Technical, Essex CM20 2JE, England (or newer versions); or (2) directly separating the mixture of optical enantiomers on a chiral chromatographic column; or (3) fractional recrystallization techniques.

[0081] It should be understood that the compounds may have tautomeric and geometric isomeric forms and that these also constitute embodiments of the present disclosure.

[0082] The present disclosure also includes isotopically labeled compounds that are identical to those set forth in formula (I) except for the fact that one or more atoms have been replaced with an atom having an atomic mass or mass number different from the atomic mass or mass number normally found in nature. Exemplary isotopes include isotopes of hydrogen, carbon, nitrogen, oxygen, phosphorus, sulfur, fluorine, and chlorine (such as, but not limited to, isotopes of hydrogen, carbon, nitrogen, oxygen, phosphorus, sulfur, fluorine, and chlorine, respectively. 2 H, 3 H, 13 C. 14 C. 15 N, 18 O. 17 O. 31 P, 32 P, 35 S, 18 F, 36 Heavier isotopes (e.g., deuterium, e.g., 2 H) may provide certain therapeutic advantages resulting in improved metabolic stability, e.g., increased in vivo half-life, or reduced dosage requirements, and therefore may be preferred in some circumstances. Positron-emitting isotopes may be incorporated into the compounds for medical imaging and positron emission tomography (PET) studies to determine receptor distribution. Suitable positron-emitting isotopes that may be incorporated into compounds of formula (I) are: 11 C. 13 N, 15 O, and 18 F. Isotopically labeled compounds of formula (I) can generally be prepared by conventional techniques known to those skilled in the art or by processes analogous to those described in the accompanying Examples, using appropriate isotopically labeled reagents in place of non-isotopically labeled reagents.

[0083] The disclosed compounds may exist as pharma- ceutically acceptable salts. The term "pharma- ceutically acceptable salts" refers to salts or zwitterions of compounds that are water- or oil-soluble or dispersible, suitable for the treatment of disorders without undue toxicity, irritation, and allergic responses, commensurate with a reasonable benefit / risk ratio, and effective for the intended use. The salts may be prepared during the final isolation and purification of the compounds, or may be prepared separately by reacting the amino group of the compounds with a suitable acid. For example, the compounds may be dissolved in a suitable solvent (e.g., but not limited to, methanol and water) and treated with at least one equivalent of an acid, such as hydrochloric acid. The resulting salts may be precipitated, isolated by filtration, and dried under reduced pressure. Alternatively, the solvent and excess acid may be removed under reduced pressure to provide the salts. Representative salts include acetate, adipate, alginate, citrate, aspartate, benzoate, benzenesulfonate, bisulfate, butyrate, camphorate, camphorsulfonate, digluconate, glycerophosphate, hemisulfate, heptanoate, hexanoate, formate, isethionate, fumarate, lactate, maleate, methanesulfonate, naphthylenesulfonate, nicotinate, oxalate, pamoate, pectinate, persulfate, 3-phenylpropionate, picrate, oxalate, maleate, pivalate, propionate, succinate, tartrate, trichloroacetate, trifluoroacetate, glutamate, para-toluenesulfonate, undecanoate, hydrochloride, hydrobromide, sulfate, phosphate, and the like. The amino groups of the compounds may also be quaternized with alkyl chlorides, bromides, and iodides, such as methyl, ethyl, propyl, isopropyl, butyl, lauryl, myristyl, stearyl, and the like.

[0084] Base addition salts can be prepared during the final isolation and purification of the disclosed compounds by reaction of the carboxyl group with a suitable base (e.g., hydroxide, carbonate, or bicarbonate) of a metal cation (e.g., lithium, sodium, potassium, calcium, magnesium, or aluminum) or an organic primary, secondary, or tertiary amine. Quaternary amine salts can be prepared, such as those derived from methylamine, dimethylamine, trimethylamine, triethylamine, diethylamine, ethylamine, tributylamine, pyridine, N,N-dimethylaniline, N-methylpiperidine, N-methylmorpholine, dicyclohexylamine, procaine, dibenzylamine, N,N-dibenzylphenethylamine, 1-ephenamine, and N,N'-dibenzylethylenediamine, ethylenediamine, ethanolamine, diethanolamine, piperidine, piperazine, and the like.

[0085] The compounds may be synthesized according to a variety of methods, including those shown in the Examples. The reaction conditions and reaction times for each individual step may vary depending on the specific reactants employed and the substituents present in the reactants used. Specific procedures are provided in the Examples section. The reactions may be worked up in a conventional manner, for example by removing the solvent from the residue, and further purified according to methodologies generally known in the art (for example, but not limited to, crystallization, distillation, extraction, trituration, and chromatography). Unless otherwise indicated, starting materials and reagents are either commercially available or can be prepared by one skilled in the art from commercially available materials using methods described in the chemical literature. If the starting materials are not commercially available, they can be prepared by procedures selected from standard organic chemistry techniques, procedures analogous to the synthesis of known structurally similar compounds, or procedures analogous to those described in the Schemes or Synthetic Examples sections above.

[0086] Routine experimentation (e.g., proper manipulation of reaction conditions, reagents, and order of synthetic pathways, protection of any chemical functional groups that are not compatible with the reaction conditions, and deprotection at suitable points in the reaction sequence of the method) is within the scope of the present disclosure. Suitable protecting groups and methods for protecting and deprotecting various substituents using such suitable protecting groups are well known to those skilled in the art; examples of this can be found in PGM Wuts and TW Greene, in Greene's book titled Protective Groups in Organic Synthesis (4th ed.), John Wiley & Sons, NY (2006) (incorporated herein by reference in its entirety). Synthesis of the compounds of the present disclosure can be accomplished by methods similar to those described in the above synthetic schemes and specific examples.

[0087] The optically active forms of the disclosed compounds can be obtained by carrying out one of the procedures described herein using, if necessary, optically active starting materials (prepared, for example, by asymmetric induction of a suitable reaction step), or by resolving a stereoisomeric mixture of the compound or intermediate using standard procedures (for example, chromatographic separation, recrystallization, or enzymatic resolution).

[0088] Similarly, if a pure geometric isomer of a compound is required, it can be obtained by carrying out one of the procedures described above using a pure geometric isomer as a starting material, or by resolving a mixture of geometric isomers of the compound or intermediates using standard procedures such as chromatographic separation.

[0089] It can be understood that the synthetic schemes and specific examples described are illustrative and should not be construed as limiting the scope of the disclosure as defined in the appended claims. All alternatives, modifications, and equivalents of the synthetic methods and specific examples are intended to be within the scope of the claims.

[0090] 3. Composition The present disclosure relates to a compound of formula (II): [ka] or a pharma- ceutically acceptable salt thereof, wherein: W is an amino acid; X is O, NR 2 , S, and a bond; R 2 is hydrogen, C1-C6 alkyl, C2-C6 alkenyl, C2-C6 alkynyl, C3-C6 cycloalkyl, or halo-C1-C6-alkyl; L is C8-C 80 Alkyl, C8-C 80 Alkenyl, C8-C 80 Alkynyl, C8-C 80 Heteroalkyl, C8-C 80 Heteroalkenyl, and C8-C 80 heteroalkynyl, each of which is optionally substituted with one or more substituents selected from hydroxy and amino.

[0091] The term "amino acid" as used herein refers to any and all amino acids, including naturally occurring amino acids (e.g., α-amino acids), non-natural amino acids, modified amino acids, and non-natural amino acids. In some embodiments, the term "amino acid" refers to any molecule that contains both an amine-terminal functional group and a carboxyl-terminal functional group. In some embodiments, the term "amino acid" refers to any amino acid-like compound that is similar in structure and / or overall shape to a naturally occurring amino acid. An amino acid can be a D- or L-amino acid. Natural amino acids include those found in nature, such as, for example, amino acids that are linked into peptide chains to form the building blocks of countless proteins. These are primarily L-stereoisomers, although a small number of D-amino acids occur in bacterial envelopes and some antibiotics. "Non-standard" natural amino acids include, for example, pyrrolysine (present in methanogens and other eukaryotes), selenocysteine ​​(present in many non-eukaryotes as well as most eukaryotes), and N-formylmethionine (encoded by the start codon AUG in bacteria, mitochondria, and chloroplasts). "Unnatural" or "non-natural" amino acids are non-proteinogenic amino acids (e.g., amino acids that are not naturally encoded or found in the genetic code) that are either naturally occurring or chemically synthesized. Over 140 unnatural amino acids are known, with thousands more possible combinations. Examples of "unnatural" amino acids include the β-amino acids (β 3 and β 2 ), homoamino acids, proline and pyruvate derivatives, 3-substituted alanine derivatives, glycine derivatives, ring-substituted phenylalanine and tyrosine derivatives, linear core amino acids, diamino acids, D-amino acids, alpha-methyl amino acids and N-methyl amino acids. Unnatural or non-natural amino acids also include modified amino acids. "Modified" amino acids include amino acids (e.g., natural amino acids) that have been chemically modified to include a group(s) or chemical moiety that is not naturally present in an amino acid.

[0092] Reference to a particular amino acid encompasses both the covalent and non-covalent states of that amino acid. Thus, the term "amino acid" refers to the residue of an amino acid whose structural part participates in the bond, often losing an atom from the amino acid itself, for example, the equivalent of a water molecule when forming a peptide bond in a protein. The amino acid may be conjugated to the lipid moiety at any suitable functional group in the amino acid. In some embodiments, the amino acid is conjugated at the carboxy terminus. In some embodiments, the amino acid is conjugated at the carbonyl carbon atom of a carboxylic acid residue.

[0093] For the most part, the names of natural and unnatural aminoacyl residues used herein follow the naming conventions proposed by the IUPAC Organic Chemical Nomenclature Commission and the IUPAC-IUB Biochemical Nomenclature Commission, as set forth in "Nomenclature of α-Amino Acids (Recommendations, 1974)" Biochemistry, 14(2), (1975). To the extent that the names and abbreviations of amino acids and aminoacyl residues used in this specification and the appended claims differ from those suggested, they will be made clear to the reader.

[0094] Throughout this specification, naturally occurring amino acids are designated by their conventional three letter or one letter abbreviations (e.g., Ala or A for alanine, Arg or R for arginine, etc.) unless they are referred to by their full name (e.g., alanine, arginine, etc.). As used herein, the term "L-amino acid" refers to the "L" isomer of a peptide, and conversely, the term "D-amino acid" refers to the "D" isomer of a peptide (e.g., Dphe, (D)Phe, D-Phe, or D-Phe for the D isomer of phenylalanine). D The D-isomer amino acid residues can be substituted with any L-amino acid residue as long as the desired function is maintained.

[0095] For less common or non-naturally occurring amino acids, unless referred to by their full name (e.g., sarcosine, ornithine, etc.), frequently used three or four letter codes are used for the residue, including Sar or Sarc (sarcosine, e.g., N-methylglycine), Aib (α-aminoisobutyric acid), Dab (2,4-diaminobutyric acid), Dapa (2,3-diaminopropanoic acid), γ-Glu (γ-glutamic acid), Gaba (γ-aminobutyric acid), β-Pro (pyrrolidine-3-carboxylic acid), and 8Ado (8-amino-3,6-dioxaoctanoic acid), Abu (2-aminobutyric acid), βhPro (β-homoproline), βhPhe (β-homophenylalanine) and Bip (β,β diphenylalanine), and Ida (iminodiacetic acid).

[0096] In some embodiments, W is tryptophan, proline, lysine, histidine, arginine, or a variant thereof. In some embodiments, W is tryptophan or a variant thereof.

[0097] In some embodiments, the compound formula (II) is a compound of formula (IIa) and W is 1-methyl-D-tryptophan. In some embodiments, the compound of formula (IIa) is: [ka] or a pharma- ceutically acceptable salt thereof.

[0098] In some embodiments, X is O or a bond. In some embodiments, X is O. In some embodiments, X is a bond. In some embodiments, X is NR 2 and R 2 is selected from hydrogen and C1-C6 alkyl. In some embodiments, X is NH. In some embodiments, X is NR 2 and R 2 In some embodiments, X is S.

[0099] In some embodiments, L is a lipid moiety having at least 8 carbon atoms, and the lipid moiety can be derived from any suitable lipid (e.g., fatty alcohol, fatty acid, phospholipid, steroid, or synthetic lipid). In some embodiments, the lipid moiety is derived from a lipid having a functional group (e.g., a hydroxyl group, a carboxylic acid group, or an amino group), and the lipid moiety is attached to the compound of formula (II) via the functional group. For example, in some embodiments, L is a lipid moiety having at least 8 carbon atoms, and in such embodiments, the group X in formula (I) is derived from a functional group; for example, when the lipid moiety is derived from an aliphatic alcohol, X is O.

[0100] In some embodiments, L is C-C 80 Alkyl, C8-C 80 Alkenyl, C8-C 80 Alkynyl, C8-C 80 Heteroalkyl, C8-C 80 Heteroalkenyl, and C8-C 80 heteroalkynyl, each of which is optionally substituted with one or more substituents selected from hydroxy and amino.

[0101] For example, in some embodiments, L is C-C 80 Alkyl and C8-C 80 In some embodiments, L is selected from C-C alkenyl. 40 Alkyl and C8-C 40 In some embodiments, L is selected from C 12 -C 40 Alkyl and C 12 -C 40In some such embodiments, L is derived from a saturated or unsaturated fatty alcohol. In some embodiments, L is derived from: linoleyl alcohol ((9Z,12Z)-octadeca-9,12-dien-1-ol), myristyl alcohol (1-tetradecanol), palmitoleyl alcohol ((Z)-hexadec-9-en-1-ol), oleyl alcohol ((Z)-octadec-9-en-1-ol), elaidyl alcohol (trans-9-octadecenol), cis-butyl alcohol (cis-butyl alcohol ... Xenyl alcohol (cis-11-octadecenol), gadoleyl alcohol ((Z)-icos-9-en-1-ol), 11-eicosenol, erucyl alcohol (cis-13-docosenol), 15-tetracosen-1-ol, eicosadienyl alcohol (icosa-11,14-dien-1-ol), linolenyl alcohol ((9Z,12Z,15Z)-9,12,15-octadecatrien-1-ol), ol), γ-linolenyl alcohol ((6E,9E,12E)-octadeca-6,9,12-trien-1-ol), eleostearyl alcohol (octadeca-9,11,13-trien-1-ol), icos-5,8,11-trien-1-ol, icos-13-en-1-ol, icos-11,14-17-trien-1-ol, octadeca-6,9,12,15-tetraen-1-ol, arachidonic acid 4E,6Z-Hexadecadien-1-ol, icosa-5,8,11,14,17-pentaen-1-ol, docosahexaenoyl alcohol (docosa-4,7,10,13,16,19-hexaen-1-ol), docosa-7,10,13,16,19-pentaen-1-ol, tetracosa-6,9,12,15,18,21-Hexaen-1-ol, Capryl Alcohol (1-Octanol), Pelargonic Alcohol (1-Nonanol), Decyl Alcohol (1-Decanol), Undecyl Alcohol (1-Undecanol), Lauryl Alcohol (1-Dodecanol), Tridecyl Alcohol (1-Tridecanol), Myristyl Alcohol (1-Tetradecanol), Pentadecyl Alcohol (1-Pentadecanol), Cetyl Alcohol (1-Hexadecanol), Palmitoleic Alcohol (cis-9-Hexadecen-1-ol), Heptadecyl Alcohol (1-n-Heptadecanol), Stearyl Alcohol (1-Octadecanol), Oleyl Alcohol (1-Octadecenol), Nonadecanyl Alcohol (1-Nonadecanol), Arachidyl Alcohol (1-Eicosanol), Heneicosyl Alcohol 1-Heneicosanol, Behenyl Alcohol, 1-Docosanol, Erucyl Alcohol, 1-Tricosyl Alcohol, 1-Tetracosanol, Pentacosyl Alcohol, 1-Pentacosanol, Seryl Alcohol, 1-Heptacosanol, Montanyl Alcohol, 1-Octacosanol, 1-Nonacosanol, Myricyl Alcohol, 1-Triacontanol, 1-Hentriacontanol, 1-Dotriacontanol, Laceryl Alcohol, 1-Tritriacontanol, Gedil Alcohol, 1-Tetratriacontanol, 1-Hexatriacontanol, 1-Heptatriacontanol, 1-Octatriacontanol, Nonatriacontan-1-ol, or 1-Tetracontanol.

[0102] In some embodiments, L is C-C 80 Heteroalkyl, C8-C 80 Heteroalkenyl, and C8-C 80 In such embodiments, L may be derived from a lipid that includes one or more heteroatom groups (e.g., -O-, -NH-, -C(O)-, etc., or combinations thereof (e.g., -C(O)O- groups).

[0103] For example, in some embodiments, L has the formula (A): [ka] (In the formula, n is 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, or 40; R a and R b are each independently C6-C 40 Alkyl, C6-C 40 Alkenyl, C6-C 40 Heteroalkyl and C6-C 40 heteroalkenyl).

[0104] For example, in some embodiments, n is 1, 2, 3, 4, 5, 6, 7, or 8. In some embodiments, n is 2, 3, or 4. In some embodiments, n is 2. In some embodiments, n is 3. In some embodiments, n is 4.

[0105] In some embodiments, R a and R b are each independently C6-C 40 Alkyl and C6-C 40 For example, in some embodiments, R a and R bare each independently selected from the following: n-hexyl, n-heptyl, n-octyl, n-nonyl, n-decyl, undecyl, dodecyl, tridecyl, tetradecyl, pentadecyl, hexadecyl, heptadecyl, octadecyl, nonadecyl, icosyl, henicosyl, docosyl, tricosyl, tetracosyl, pentacosyl, hexacosyl, heptacosyl, octacosyl, nonacosyl, triacontyl, hentriacontyl, dotriacontyl, tritriacontyl, tetratriacontyl, pentatriacontyl, pentyl, hexatriacontyl, heptatriacontyl, octatriacontyl, nonatriacontyl, tetracontyl, linoleyl ((9Z,12Z)-octadeca-9,12-dien-1-yl), palmitoleyl ((Z)-hexadec-9-en-1-yl), oleyl ((Z)-octadec-9-en-1-yl), elaidyl (trans-9-octadecenyl), cis-vaccenyl (cis-11-octadecenyl), gadoleyl ((Z)-icos-9-en-1-yl), 11-eicosenyl, elci cis-13-docosenyl, 15-tetracosen-1-yl, eicosadienyl (icosa-11,14-dien-1-yl), linolenyl ((9Z,12Z,15Z)-9,12,15-octadecatrien-1-yl), γ-linolenyl ((6E,9E,12E)-octadeca-6,9,12-trien-1-yl), eleostearyl (octadeca-9,11,13-trien-1-yl), icos-5,8,11-trien-1-yl, eicos-13-en-1-yl, icos-11,14-17- In some embodiments, R is selected from the group consisting of icosatetraen-1-yl, octadeca-6,9,12,15-tetraen-1-yl, arachidonyl ((5Z,8Z,11Z,14Z)-icosatetraen-1-yl), 4E,6Z-hexadecadien-1-yl, icosa-5,8,11,14,17-pentaen-1-yl, docosahexaenoyl (docosa-4,7,10,13,16,19-hexaen-1-yl), docosa-7,10,13,16,19-pentaen-1-yl, and tetracosa-6,9,12,15,18,21-hexaen-1-yl. a and R b are linoleyl, respectively.

[0106] In some embodiments, L has the formula (B) or (C): [ka] (In the formula, n and p are each independently 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, or 40; R a1 is C6-C 40 Alkyl and C6-C 40 alkenyl, R b is C6-C 40 Alkyl, C6-C 40 Alkenyl, C6-C 40 Heteroalkyl and C6-C 40 heteroalkenyl).

[0107] In some embodiments, n is 1, 2, 3, 4, 5, 6, 7, or 8. In some embodiments, n is 2, 3, or 4. In some embodiments, n is 2. In some embodiments, n is 3. In some embodiments, n is 4.

[0108] In some embodiments, p is 1, 2, 3, 4, 5, 6, 7, or 8. In some embodiments, p is 3, 4, 5, 6, or 7. In some embodiments, p is 3. In some embodiments, p is 4. In some embodiments, p is 5. In some embodiments, p is 6. In some embodiments, p is 7.

[0109] In some embodiments, R a1 is C6-C 24 Alkyl or C6-C 24 In some embodiments, R is alkenyl. a1is C9-C 22 Alkyl and C9-C 22 In some embodiments, R is selected from the group consisting of alkenyl and aryl. a1 are linear or branched C6, C7, C8, C9, C 10 , C 11 , C 12 , C 13 , C 14 , C 15 , C 16 , C 17 , C 18 , C 19 , C 20 , C 21 , C 22 , C 23 , or C 24 In some embodiments, R a1 are linear or branched C6, C7, C8, C9, C 10 , C 11 , C 12 , C 13 , C 14 , C 15 , C 16 , C 17 , C 18 , C 19 , C 20 , C 21 , C 22 , C 23 , or C 24 In some embodiments, R is selected from the group consisting of alkenyl and aryl. a1 is selected from linoleyl, n-nonyl, n-undecyl, henicosan-11-yl, pentadecan-7-yl, and heptadecan-9-yl.

[0110] In some embodiments, R b is C6-C 40 Alkyl and C6-C 40 For example, in some embodiments, R bis selected from the following: n-hexyl, n-heptyl, n-octyl, n-nonyl, n-decyl, undecyl, dodecyl, tridecyl, tetradecyl, pentadecyl, hexadecyl, heptadecyl, octadecyl, nonadecyl, icosyl, henicosyl, docosyl, tricosyl, tetracosyl, pentacosyl, hexacosyl, heptacosyl, octacosyl, nonacosyl, triacontyl, hentriacontyl, dotriacontyl, tritriacontyl, tetratriacontyl, pentatriacontyl, hexatriacontyl, Satriacontyl, heptatriacontyl, octatriacontyl, nonatriacontyl, tetracontyl, linoleyl ((9Z,12Z)-octadeca-9,12-dien-1-yl), palmitoleyl ((Z)-hexadec-9-en-1-yl), oleyl ((Z)-octadec-9-en-1-yl), elaidyl (trans-9-octadecenyl), cis-vaccenyl (cis-11-octadecenyl), gadoleyl ((Z)-icos-9-en-1-yl), 11-eicosenyl, erucyl (cis -13-docosenyl), 15-tetracosen-1-yl, eicosadienyl (icosa-11,14-dien-1-yl), linolenyl ((9Z,12Z,15Z)-9,12,15-octadecatrien-1-yl), γ-linolenyl ((6E,9E,12E)-octadeca-6,9,12-trien-1-yl), eleostearyl (octadeca-9,11,13-trien-1-yl), icos-5,8,11-trien-1-yl, eicos-13-en-1-yl, icos-11,14-17-trien-1-yl In some embodiments, R is selected from the group consisting of octadeca-6,9,12,15-tetraen-1-yl, arachidonyl ((5Z,8Z,11Z,14Z)-icosatetraen-1-yl), 4E,6Z-hexadecadien-1-yl, icosa-5,8,11,14,17-pentaen-1-yl, docosahexaenoyl (docosa-4,7,10,13,16,19-hexaen-1-yl), docosa-7,10,13,16,19-pentaen-1-yl, and tetracosa-6,9,12,15,18,21-hexaen-1-yl. b is linoleyl.

[0111] In some embodiments, L has the formula (D), (E), or (F): [ka] (In the formula, n, p, and q are each independently 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, or 40; R a1 and R a2 are each independently C6-C 40 Alkyl and C6-C 40 alkenyl).

[0112] In some embodiments, L has formula (D). In some embodiments, L has formula (E). In some embodiments, L has formula (F). In some embodiments, n is 1, 2, 3, 4, 5, 6, 7, or 8. In some embodiments, n is 2, 3, or 4. In some embodiments, n is 2. In some embodiments, n is 3. In some embodiments, n is 4.

[0113] In some embodiments, p and q are each independently selected from 1, 2, 3, 4, 5, 6, 7, or 8. In some embodiments, p and q are each independently selected from 3, 4, 5, 6, and 7. In some embodiments, p and q are each independently selected from 3, 5, and 7. In some embodiments, p and q are each 5. In some embodiments, p and q are each 6. In some embodiments, p and q are each 7.

[0114] In some embodiments, R a1 and R a2 are each independently C6-C 24 Alkyl and C6-C 24In some embodiments, R is selected from the group consisting of alkenyl and aryl. a1 and R a2 are each independently C9-C 22 Alkyl and C9-C 22 In some embodiments, R is selected from the group consisting of alkenyl and aryl. a1 and R a2 are each independently a linear or branched C6, C7, C8, C9, C 10 , C 11 , C 12 , C 13 , C 14 , C 15 , C 16 , C 17 , C 18 , C 19 , C 20 , C 21 , C 22 , C 23 , or C 24 In some embodiments, R a1 and R a2 are each independently a linear or branched C6, C7, C8, C9, C 10 , C 11 , C 12 , C 13 , C 14 , C 15 , C 16 , C 17 , C 18 , C 19 , C 20 , C 21 , C 22 , C 23 , or C 24 In some embodiments, R is selected from the group consisting of alkenyl and aryl. a1 and R a2 are each independently selected from linoleyl, n-nonyl, n-undecyl, henicosan-11-yl, pentadecan-7-yl, and heptadecan-9-yl.

[0115] In some embodiments, L is derived from a steroid, for example, in some embodiments, L is derived from cholesterol, beta-sisterol, or BHEM cholesterol.

[0116] In some embodiments, L is selected from: [ka] [ka] [ka]

[0117] The composition may take the form of liposomes, lipid nanoparticles, micelles, etc. The lipid composition manufacturing method includes, for example, lipid film hydration (optionally combined with sonication or extrusion), solvent evaporation (e.g., ethanol injection, ether injection, or reverse phase evaporation), solvent diffusion method, high temperature homogenization process, detergent removal method, or combinations thereof. Natural or synthetic vesicle-forming lipids, or combinations thereof, can be used. One or more vesicle-forming lipids can be selected from bialiphatic chain lipids (e.g., phospholipids); diglycerides; bialiphatic glycolipids; single lipids (e.g., sphingomyelin or glycosphingolipids); steroid lipids; hydrophilic polymer-derivatized lipids; or mixtures thereof.

[0118] The lipid compositions of the present disclosure may comprise one or more cationic and / or ionizable lipids, phospholipids, neutral or non-cationic lipids, polyethylene glycol (PEG) lipid conjugates, and / or structured lipids.

[0119] Cationic lipids and / or ionizable lipids include, for example, amine-containing lipids, which can be readily protonated and have a positive or partial positive charge at physiological pH, since they have pKa values ​​between pH 5 and 8. The polar head groups of the cationic lipids preferably include amine derivatives, such as primary amines, secondary amines, and / or tertiary amines, quaternary ammonium, various combinations of amines, amidinium salts, or guanidine and / or imidazole groups, as well as pyridinium, piperazine, and amino acid head groups (e.g., lysine, arginine, ornithine, and / or tryptophan). Cationic lipids include, but are not limited to, 1,2-dimyristoyl-sn-glycero-3-ethylphosphocholine (DMEPC), 1,2-di-O-octadecenyl-3-trimethylammonium propane (DOTMA) and / or 1,2-dioleoyl-3-trimethylammonium propane (DOTAP), 1,2-dimyristoyl-3-trimethylammonium propane (DMTAP), 2,3-di(tetradecoxy)propyl-(2-hydroxyethyl)dimethylazanium bromide (DMRIE), didodecyl(dimethyl)ammonium bromide (DDAB), 1,2-dioleyloxypropyl-3-dimethylhydroxyethylammonium bromide (DORIE), 3β-[N-(N,N′-dimethylaminoethane)carbamoyl]cholesterol (DC-Chol), or dioleyl ether phosphatidylcholine (DOEPC). Ionizable lipids include, but are not limited to, 1,2-dioleyloxy-3-dimethylaminopropane (DODMA).

[0120] In some embodiments, the disclosed compounds are incorporated into lipid compositions comprising one or more phospholipids. The phospholipid comprises a phospholipid moiety and one or more fatty acid moieties. The phospholipid moiety may comprise, but is not limited to, phosphatidylcholine, phosphatidylethanolamine, phosphatidylglycerol, phosphatidylserine, phosphatidic acid, 2-lysophosphatidylcholine, and sphingomyelin. The fatty acid moiety may comprise, but is not limited to, lauric acid, myristic acid, myristoleic acid, palmitic acid, palmitoleic acid, stearic acid, oleic acid, linoleic acid, alpha-linolenic acid, erucic acid, phytanic acid, arachidic acid, arachidonic acid, eicosapentaenoic acid, behenic acid, docosapentaenoic acid, and docosahexaenoic acid.

[0121] Phospholipids suitable for use in the composition may include, but are not limited to, phosphatidylglycerol (PG), (e.g., dimyristoylphosphatidylglycerol (DMPG) and 1,2-dioleoyl-sn-glycero-3-phospho-rac-(1-glycerol) sodium salt (DOPG)); phosphatidylcholine (PC), (e.g., egg yolk phosphatidylcholine, dimyristoylphosphatidylcholine (DMPC), 1,2-distearoyl-sn-glycero-3-phospho-rac-(1-glycerol) sodium salt (DOPG)); 1,2-dilinoleoyl-sn-glycero-3-phosphocholine (DLPC), 1,2-dioleoyl-sn-glycero-3-phosphocholine (DOPC), 1,2-dipalmitoyl-sn-glycero-3-phosphocholine (DPPC), 1,2-diundecanoyl-sn-glycero-phosphocholine (DUPC), 1-palmitoyl-2-oleoyl-sn-glycero-3-phosphocholine (POPC), 1,2-di-O-octadecenyl-sn-glycero-3-phosphocholine (18:0 diether PC), 1-oleoyl-2-cholesterylhemisuccinoyl-sn-glycero-3-phosphocholine (OChemsPC), 1-hexadecyl-sn-glycero-3-phosphocholine (C16 Lyso PC), 1,2-dilinolenoyl-sn-glycero-3-phosphocholine, 1,2-didocosahexaenoyl-sn-glycero-3-phosphocholine, 1,2-diarachidonoyl-sn-glycero-3-phosphocholine; phosphatidylethanolamines (PE) (e.g., 1,2-dioleoyl-sn-glycero-3-phosphoethanolamine (DOPE), 1,2-diphytanoyl-sn-glycero-3-phosphoethanolamine (ME 16.0 PE), 1,2-distearoyl-sn-glycero-3-phosphoethanolamine, 1,2-dilinoleoyl-sn-glycero-3-phosphoethanolamine, 1,2-dilinolenoyl-sn-glycero-3-phosphoethanolamine, 1,2-diarachidonoyl-sn-glycero-3-phosphoethanolamine, 1,2-didocosahexaenoyl-sn-glycero-3-phosphoethanolamine); phosphatidic acid (PA); phosphatidylinositol (PI); phosphatidylserine (PS); and sphingomyelin (SM).

[0122] In some embodiments, the lipid composition comprises a structured lipid. In some embodiments, the structured lipid is a sterol. The sterol may comprise cholesterol, fecosterol, ergosterol, campesterol, sitosterol, stigmasterol, brassicasterol, or sterol esters (e.g., cholesteryl hemisuccinate, cholesteryl sulfate, ursolic acid, α-tocopherol) or any other derivative of cholesterol.

[0123] In some embodiments, the lipid composition comprises a polyethylene glycol (PEG)-lipid conjugate. The PEG-lipid conjugate may include, but is not limited to, a PEG-modified phosphatidylethanolamine, a PEG-modified phosphatidic acid, a PEG-modified ceramide, a PEG-modified dialkylamine, a PEG-modified diacylglycerol, a PEG-modified dialkylglycerol, and mixtures thereof. For example, the PEG lipid may be a PEG-DMG (1,2-dimyristoyl-rac-glycero-3-methoxypolyethylene glycol), PEG-c-DOMG (R-3-[(ω-methoxypoly(ethylene glycol)2000)carbamoyl)]-1,2-dimyristyloxypropyl-3-amine), PEG-DMA (PEG-dimethacrylate), PEG-DLPE (1,2-didodecanoyl-sn-glycero-3-phosphoethanolamine-PEG), PEG-DMPE (PEG-1,2-dimyristoyl-sn-glycero-3-phosphoethanolamine), PEG-DPPC (PEG-dipalmitoylphosphatidylcholine), PEG-N,N-di(tetradecyl)acetamide, or PEG-DSPE (1,2-distearoyl-sn-glycero-3-phosphoethanolamine-poly(ethylene glycol)) lipid. In some embodiments, the lipid nanoparticles comprise PEG-DMG and / or PEG-N,N-di(tetradecyl)acetamide.

[0124] The lipid structures described herein may also contain other components that are typically used in the formation of vesicles (e.g., for stabilization). Examples of such other components include, but are not limited to, fatty alcohols, fatty acids, and / or any other pharma- ceutically acceptable excipients that can affect surface charge, membrane fluidity, and aid in the incorporation of lipids into lipid assemblies.

[0125] The lipid composition may also be targeted, e.g., contain one or more targeting moieties or biodistribution modifiers on the surface. The targeting moiety may be any agent capable of specifically binding or interacting with a desired target, and is generally known in the art, e.g., a ligand (e.g., folic acid, a protein, an antibody, or an antibody fragment, etc.). In some embodiments, the targeting moiety is an immune cell epitope (e.g., B cell and T cell epitopes). In selected embodiments, the targeting moiety comprises one or more epitopes from microbial agents (e.g., Clostridioides difficile, Bacillus anthracis, clostridium botulinum, Helicobacter pylori, Rotavirus sp., Coronaviridae).

[0126] The lipid composition may have any structure (e.g., a structure having an interior space separated from the external medium by one or more lipid bilayers) or any microcapsule having a semipermeable membrane with a lipophilic central portion that separates the interior of the membrane. In some embodiments, the lipid composition may comprise unilamellar liposomes having a single lipid layer. In some embodiments, the lipid composition may comprise liposomes having a lipid bilayer. In some embodiments, the lipid composition comprises micelles.

[0127] 4. Active drugs Any of the compositions disclosed herein may further comprise at least one active agent. In some embodiments, the at least one active agent comprises a nucleic acid (mRNA, aptamer, antisense oligonucleotide, ribozyme nucleic acid, interfering RNA, antisense, and antigene nucleic acid), a protein, an immunomodulator, a chemotherapeutic drug, a steroid, an analgesic drug, an antibacterial drug, or a combination thereof. In some embodiments, the at least one active agent is encapsulated in a liposome, lipid nanoparticle, or micelle in a lipid composition.

[0128] In some embodiments, at least one active agent comprises a nucleic acid (e.g., ribonucleic acid or deoxyribonucleic acid). Exemplary nucleic acids used in accordance with the present disclosure include, but are not limited to, one or more of deoxyribonucleic acid (DNA), ribonucleic acid (RNA) (e.g., messenger mRNA (mRNA)), hybrids thereof, RNAi inducers, RNAi agents, siRNA, shRNA, miRNA, antisense RNA, ribozymes, catalytic DNA, RNA that induces triple helix formation, aptamers, vectors, and the like. In some embodiments, the nucleic acid encodes an antigen or a functional fragment thereof.

[0129] In some embodiments, at least one active agent is RNA. The RNA useful in the compositions and methods described herein can be selected from the group consisting of, but not limited to: shortmer, antagomir, antisense RNA, ribozyme, small interfering RNA (siRNA), asymmetric interfering RNA (aiRNA), microRNA (miRNA), dicer substrate RNA (dsRNA), small hairpin RNA (shRNA), transfer RNA (tRNA), messenger RNA (mRNA), and mixtures thereof.

[0130] In certain embodiments, at least one active agent is an mRNA.The mRNA can code for any polypeptide of interest, including any naturally occurring or non-naturally occurring or otherwise modified polypeptide.The polypeptide coded by the mRNA can be of any size and have any secondary structure or activity.In some embodiments, the polypeptide coded by the mRNA can have a therapeutic effect when expressed in a cell.

[0131] In other embodiments, at least one active agent is an siRNA. The siRNA can selectively knock down or down regulate the expression of a gene of interest. For example, the siRNA can be selected to silence a gene associated with a particular disease, disorder, or condition when a nanoparticle composition comprising the siRNA is administered to a subject in need thereof. The siRNA can include a sequence that is complementary to the mRNA sequence that codes for the gene or protein of interest. In some embodiments, the siRNA can be an immunomodulatory siRNA.

[0132] In some embodiments, at least one active agent is shRNA or its encoding vector or plasmid.shRNA can be produced in target cell when appropriate construct is delivered to nucleus.The structure and mechanism related to shRNA are well known in the related art.

[0133] In some embodiments, at least one active agent is an immunomodulator. Exemplary immunomodulators include: signal transducer and activator of transcription 3 (Stat3) inhibitors and analogs thereof (e.g., SM-36 and analogs thereof); toll-like receptor (TLR) agonists and analogs thereof (e.g., imiquimod, resiquimod, sergantolimod, gardiquimod, SM-360320, TMX-101, TMX-202, TMX-302, TMX-306, GSK2245035, CL097, 852A, AZD-8848, DSP-3025, GS-9620, RO7020531, RO6871765, ANA773, DSP-0509, NJH395, BNT411, TQ-A3334, JNJ-4964, LHC165, CV8102, VTX-1463, VTX-2337, IMO-8400, IMO-3100, IRS-954, and their analogs); and statins or other lipid-lowering drugs and their analogs (e.g., atorvastatin, pravastatin, fluvastatin, simvastatin, lovastatin, mevastatin, pitavastatin, rosuvastatin, and their analogs).

[0134] In some embodiments, the at least one active agent comprises at least one chemotherapeutic agent. As used herein, the term "chemotherapeutic agent" or "anticancer agent" includes any small molecule or other agent used in the treatment or prevention of cancer. Chemotherapeutic agents include, but are not limited to, cyclophosphamide, methotrexate, 5-fluorouracil, doxorubicin, docetaxel, daunorubicin, bleomycin, vinblastine, dacarbazine, cisplatin, paclitaxel, raloxifene hydrochloride, tamoxifen citrate, abemacirb, afinitor (everolimus), alpelisib, anastrozole, pamidronate, anastrozole, exemestane, capecitabine, epirubicin hydrochloride, eribulin mesylate, toremifene, fulvestrant, letrozole, gemcitabine, goserelin, ixabepilone, emtansine, lapatinib, olaparib, megestrol, neratinib, palbociclib, ribociclib, talazoparib, thiotepa, toremifene, methotrexate, and tucatinib.

[0135] In some embodiments, at least one active agent comprises an antimicrobial agent (eg, an antiviral or antibacterial agent). In some embodiments, the antibacterial agent is an antiviral agent (e.g., abacavir, acyclovir, adefovir, amantadine, amprenavir, atazanavir, baloxavir marboxil, bictarvy, boceprevir, brevirtide, cidofovir, cobicistat, daclatasvir, darunavir, delavirdine, didanosine, docosanol, dolutegravir, doravirine, edoxudine, efavirenz, elvitegravir, emtricitabine, enfuvirtide, entecavir, etravirine, famciclovir, fomivirsen, fosamprenavir, foscarnet, ganciclovir, ibacitabine, ibalizumab, idoxuridine, imiquimod, imunovir, indinavir, lamivudine, letermovir, lopinavir, loviride, maraviroc, mevir, serovar ... tisane, moroxydine, nelfinavir, nevirapine, nexavir, nitazoxanide, oseltamivir, penciclovir, peramivir, penciclovir, peramivir, pleconaril, podophyllotoxin, raltegravir, remdesivir, ribavirin, rilpivirine, rimantadine, lintatolimod, ritonavir, saquinavir, simeprevir, sofosbuvir, stavudine, taribavirin, telaprevir, telbivudine, tenofovir (e.g., tenofovir alafenamide or tenofovir disoproxil), tipranavir, trifluridine, tromantadine, umifenovir, valacyclovir, valganciclovir, vicriviroc, vidarabine, zalcitabine, zanamivir, and zidovudine, and combinations thereof.

[0136] In some embodiments, the antibacterial agent is an antimicrobial agent. Examples of antibacterial agents include sulfonamides, amphenicols, spectinomycin, trimethoprim, glycylcyclines, macrolides (e.g., erythromycin, clarithromycin, azithromycin, roxithromycin), oxazolidinones (e.g., linezolid), tetracyclines (e.g., doxycycline, tetracycline, minocycline), β-lactams (e.g., penicillin, methicillin, cloxacillin), carbapenems (e.g., imipenem, meropenem, aztreonam), aminoglycosides (e.g., gentamicin, tobramycin, amikacin), quinolones and fluoroquinolones (e.g., levofloxacin, ciprofloxacin, moxifloxacin), glycopeptides (e.g., vancomycin), and polymyxins (e.g., polymyxin, colistin).

[0137] 5. Pharmaceutical Compositions The lipid composition can be incorporated into a pharma- ceutical acceptable composition. Pharmaceutical compositions and formulations may include a pharma- ceutical acceptable carrier. As used herein, the term "pharma- ceutical acceptable carrier" refers to a non-toxic, inert solid, semi-solid or liquid filler, diluent, encapsulating material, surfactant, cyclodextrin, or any type of formulation auxiliary. Some examples of substances that may function as pharma- ceutically acceptable carriers include sugars (e.g., but are not limited to, lactose, glucose, and sucrose); starches (e.g., but are not limited to, corn starch and potato starch); cellulose and its derivatives (e.g., but are not limited to, sodium carboxymethylcellulose, ethylcellulose, and cellulose acetate); powdered tragacanth; malt; gelatin; talc; excipients (e.g., but are not limited to, cocoa butter and suppository wax); oils (e.g., but are not limited to, peanut oil, cottonseed oil, safflower oil, sesame oil, olive oil, corn oil, and soybean oil); surfactants (e.g., but are not limited to, Cremophor EL, Cremophor RH 60, Solutol HS 15, and Polysorbate 80); Chlodextrins (for example, but not limited to, alpha-CD, beta-CD, gamma-CD, HP-beta-CD, SBE-beta-CD); glycols (for example, but not limited to, propylene glycol); esters (for example, but not limited to, ethyl oleate and ethyl laurate); agar; buffers (for example, but not limited to, magnesium hydroxide and aluminum hydroxide); alginic acid; pyrogen-free water; isotonic saline; Ringer's solution; ethyl alcohol, and phosphate buffers, as well as other non-toxic compatible lubricants (for example, but not limited to, sodium lauryl sulfate and magnesium stearate), as well as coloring agents, release agents, coating agents, sweetening agents, flavoring and perfuming agents, preservatives, and antioxidants may also be present in the composition, at the discretion of the formulator.

[0138] The route by which the composition is administered and the form of the composition will determine the type of carrier to be used. The composition may be in a variety of forms suitable for, for example, systemic administration (e.g., oral, rectal, nasal, sublingual, buccal, implant, or parenteral injection) or local administration (e.g., transdermal, pulmonary, nasal, otic, ocular, liposome delivery system, or iontophoresis).

[0139] Carriers for systemic administration typically include at least one of a diluent, lubricant, binder, disintegrant, colorant, flavorant, sweetener, antioxidant, preservative, glidant, solvent, suspending agent, wetting agent, surfactant, cyclodextrin, combinations thereof, etc. All carriers are optional in the composition.

[0140] Suitable diluents include sugars (e.g., glucose, lactose, dextrose, and sucrose); diols (e.g., propylene glycol); calcium carbonate; sodium carbonate; sugar alcohols (e.g., glycerin, mannitol, and sorbitol). The amount of diluent(s) in a systemic or topical composition is typically about 50 to about 90%.

[0141] Suitable lubricants include silica, talc, stearic acid and its magnesium and calcium salts, calcium sulfate; and liquid lubricants such as polyethylene glycols, and vegetable oils such as peanut oil, cottonseed oil, sesame oil, olive oil, corn oil, and theobroma oil. The amount of lubricant(s) in a systemic or topical composition is typically about 5 to about 10%.

[0142] Suitable binders include polyvinylpyrrolidone, magnesium aluminum silicate, starches (e.g., corn starch and potato starch), gelatin, tragacanth, and cellulose and its derivatives (e.g., sodium carboxymethylcellulose, ethylcellulose, methylcellulose, microcrystalline cellulose, and sodium carboxymethylcellulose). The amount of binder(s) in the systemic composition is typically about 5 to about 50%.

[0143] Suitable disintegrants include agar, alginic acid and its sodium salt, effervescent mixtures, croscarmelose, crospovidone, sodium carboxymethyl starch, sodium starch glycolate, clays, and ion exchange resins. The amount of disintegrant(s) in a systemic or topical composition is typically about 0.1 to about 10%.

[0144] Suitable coloring agents include colorants such as the FD&C dyes. If used, the amount of coloring agent in a systemic or topical composition is typically about 0.005 to about 0.1%.

[0145] Suitable flavorings include menthol, peppermint, and fruit flavors. The amount of flavoring(s) used in a systemic or topical composition is typically from about 0.1 to about 1.0%.

[0146] Suitable sweeteners include aspartame and saccharin. The amount of sweetener(s) in a systemic or topical composition is typically from about 0.001 to about 1%.

[0147] Suitable antioxidants include butylated hydroxyanisole ("BHA"), butylated hydroxytoluene ("BHT"), and vitamin E. The amount of antioxidant(s) in a systemic or topical composition is typically from about 0.1 to about 5%.

[0148] Suitable preservatives include benzalkonium chloride, methylparaben, and sodium benzoate. The amount of preservative(s) in a systemic or topical composition is typically about 0.01 to about 5%.

[0149] Suitable glidants include silicon dioxide. The amount of lubricant(s) in a systemic or topical composition is typically about 1 to about 5%.

[0150] Suitable solvents include water, isotonic saline, ethyl oleate, glycerin, hydroxylated castor oil, alcohol (e.g., ethanol), dimethylsulfoxide, N-methyl-2-pyrrolidone, dimethylacetamide, and phosphate (or other suitable buffer). The amount of solvent(s) in a systemic or topical composition is typically from about 0 to about 100%.

[0151] Suitable suspending agents include AVICEL RC-591 (FMC Corporation, Philadelphia, Pa.) and sodium alginate. The amount of suspending agent(s) in a systemic or topical composition is typically about 1 to about 8%.

[0152] Suitable surfactants include lecithin, polysorbate 80, and sodium lauryl sulfate, as well as TWEENS (Atlas Powder Company, Wilmington, Del.). Suitable surfactants include those disclosed in: CTFA Cosmetic Ingredient Handbook, 1992, pp. 587-592; Remington's Pharmaceutical Sciences, 15th Ed. 1975, pp. 335-337; and McCutcheon's Volume 1, Emulsifiers & Detergents, 1994, North American Edition, pp. 236-239. The amount of surfactant(s) in a systemic or topical composition is typically about 0.1% to about 5%.

[0153] Suitable cyclodextrins include alpha-CD, beta-CD, gamma-CD, hydroxypropyl betadex (HP-beta-CD), and sulfobutyl ether beta-cyclodextrin (SBE-beta-CD). The amount of cyclodextrin in a systemic or topical composition is typically about 0% to about 40%.

[0154] Compositions for oral administration can be in liquid form. For example, suitable liquid forms include aqueous solutions, emulsions, suspensions, solutions reconstituted from non-effervescent granules, suspensions reconstituted from non-effervescent granules, effervescent preparations reconstituted from effervescent granules, elixirs, tinctures, syrups, etc. Liquid compositions for oral administration usually include the disclosed compounds and carriers (i.e. carriers selected from diluents, colorants, flavorants, sweeteners, preservatives, solvents, suspending agents, and surfactants). Oral liquid compositions preferably include one or more components selected from colorants, flavorants, and sweeteners.

[0155] Other compositions useful for achieving systemic delivery of the subject compound include sublingual, buccal, and nasal dosage forms. Such compositions usually include one or more of soluble fillers (e.g., diluents including sucrose, sorbitol, and mannitol) and binders (e.g., acacia, microcrystalline cellulose, carboxymethylcellulose, and hydroxypropylmethylcellulose). Such compositions may further include lubricants, colorants, flavorings, sweeteners, antioxidants, and glidants.

[0156] The amount of carrier used in conjunction with the disclosed compositions is sufficient to provide a practical amount of the composition for administration per unit dose of compound. Techniques and compositions for preparing dosage forms useful in the methods of the present invention are described in the following references: Modern Pharmaceutics, Chapters 9 and 10, Banker & Rhodes, eds. (1979); Lieberman et al., Pharmaceutical Dosage Forms: Tablets (1981); and Ansel, Introduction to Pharmaceutical Dosage Forms, 2nd Ed., (1976).

[0157] The carrier may comprise a single component or a combination of two or more components.

[0158] 6. Vaccines The composition may also be used in a vaccine. The vaccine comprises the disclosed composition and an antigen or a nucleic acid encoding the same. Suitable antigens include microbial pathogens, bacteria, viruses, proteins, glycoproteins, lipoproteins, peptides, glycopeptides, lipopeptides, toxoids, carbohydrates, and tumor-specific antigens. A mixture of two or more antigens may be used.

[0159] Antigens can be derived from and / or isolated from essentially any desired source, depending on the infectious disease, autoimmune disease, condition, cancer, pathogen, or disease to be treated with the particular vaccine composition.

[0160] The vaccines described herein may be capable of providing immunity against one or more conditions associated with an infectious disease (e.g., but not limited to, influenza, measles, human papilloma virus (HPV), rabies, meningitis, whooping cough, tetanus, plague, hepatitis, and tuberculosis) and may include antigens and / or epitopes from an infectious disease, or nucleic acids encoding them.

[0161] The vaccines described herein may also induce an immune response against cancer cells and may include antigens, epitopes, and / or neoepitopes, or portions thereof, or nucleic acids encoding antigens, epitopes, and / or neoepitopes from tumor cells. Tumor antigens are surface molecules that are differentially expressed on tumor cells compared to non-tumor tissues. Tumor antigens immunologically distinguish tumor cells from normal cells and provide targets for the diagnosis and treatment of human cancer. Tumor antigens have been characterized as membrane proteins or altered carbohydrate molecules of glycoproteins or glycolipids on the cell surface. Cancer cells often have characteristic tumor antigens on their surface (e.g., truncated epidermal growth factor, folate binding protein, epithelial mucin, melanopherin, carcinoembryonic antigen, prostate specific membrane antigen, HER2-neu), which are candidates for use in therapeutic cancer vaccines. Because tumor antigens are normal or associated with normal components of the body, the immune system often cannot mount an effective immune response against those antigens to destroy tumor cells. Illustrative cancer types in which this approach can be used include prostate cancer, colon cancer, breast cancer, ovarian cancer, pancreatic cancer, brain cancer, head and neck cancer, melanoma, leukemia, lymphoma, and the like.

[0162] In other embodiments, the antigen present in the vaccine composition is not a foreign antigen, but an autoantigen, for example, the vaccine composition is directed to an autoimmune disease. Examples of autoimmune diseases include type 1 diabetes, conventional organ-specific autoimmunity, neurological diseases, rheumatic diseases / connective tissue diseases, autoimmune cytopenias, and related autoimmune diseases. Such conventional organ-specific autoimmunity may include thyroiditis (Graves' disease + Hashimoto's disease), gastritis, adrenalitis (Addison's disease), oophoritis, primary biliary cirrhosis, myasthenia gravis, hypogonadism, hypoparathyroidism, alopecia, malabsorption syndrome, pernicious anemia, hepatitis, anti-receptor antibody disease, and vitiligo. Such neurological diseases may include schizophrenia, Alzheimer's disease, depression, hypopituitarism, diabetes insipidus, sicca syndrome, and multiple sclerosis. Such rheumatic / connective tissue diseases may include rheumatoid arthritis, systemic lupus erythematosus (SLE) or lupus, scleroderma, polymyositis, inflammatory bowel disease, dermatomyositis, ulcerative colitis, Crohn's disease, vasculitis, psoriatic arthritis, exfoliative psoriatic dermatitis, pemphigus vulgaris, Sjogren's syndrome. Other autoimmune-related diseases may include autoimmune uveoretinitis, glomerulonephritis, post-myocardial infarction cardiotomy syndrome, pulmonary hemosiderosis, amyloidosis, sarcoidosis, aphthous stomatitis, and other immune-related diseases presented herein and known in the relevant art.

[0163] In one embodiment, the antigen in the vaccine composition is a peptide, polypeptide, or immunogenic portion thereof. As used herein, an "immunogenic portion" is a portion of a protein that is recognized (e.g., specifically binds) by a B-cell and / or T-cell surface antigen receptor. Such an immunogenic portion generally comprises at least 5 amino acid residues, more preferably at least 10, and even more preferably at least 20 amino acid residues of an antigenic protein or a variant thereof.

[0164] Immunogenic portions of an antigenic polypeptide may generally be identified using well-known techniques, such as those summarized in Paul, Fundamental Immunology, 3rd ed., 243-247 (Raven Press, 1993) and references cited therein. Such techniques include screening the polypeptide for the ability to react with antigen-specific antibodies, antisera, and / or T-cell lines or clones. As used herein, antisera and antibodies are "antigen-specific" if they specifically bind to the antigen (e.g., react with the protein in an ELISA or other immunoassay and show no detectable reaction with unrelated proteins). Such antisera and antibodies may be prepared using well-known techniques. An immunogenic portion of a protein is one that reacts with such antisera and / or T cells (e.g., in an ELISA and / or T-cell reactivity assay) at a level not substantially lower than the reactivity of the full-length polypeptide. Such immunogenic portions may react within such assays at a level equal to or greater than the reactivity of the full-length polypeptide. Such screening may generally be performed using methods well known to those of skill in the art, such as those described in Harlow and Lane, Antibodies: A Laboratory Manual, Cold Spring Harbor Laboratory, 1988. For example, the polypeptide may be immobilized on a solid support and contacted with patient serum to allow binding of antibodies in the serum to the immobilized polypeptide. Unbound serum may then be removed, e.g., by removing sera from the patient. 125 I-labeled protein A is used to detect bound antibody.

[0165] Peptide and polypeptide antigens can be prepared using any of a variety of well-known techniques. Recombinant polypeptides encoded by DNA sequences can be readily prepared from isolated DNA sequences using any of a variety of expression vectors known to those skilled in the art. Expression can be achieved in any suitable host cell that has been transformed or transfected with an expression vector containing a DNA molecule encoding a recombinant polypeptide. Suitable host cells include prokaryotes, yeast, and higher eukaryotic cells (e.g., mammalian cells and plant cells). Preferably, the host cells used are E. coli, yeast, or mammalian cell lines (e.g., COS or CHO).

[0166] Portions and other variants of protein antigens having fewer than about 100 amino acids, generally fewer than about 50 amino acids, can also be produced by synthetic means using techniques well known to those of skill in the art. For example, such polypeptides can be synthesized using any of the commercially available solid-phase methods, such as the Merrifield solid-phase synthesis method, in which amino acids are added sequentially to a growing amino acid chain. See, Merrifield, J. Am. Chem. Soc. 85:2149-2146, 1963. Automated polypeptide synthesizers are commercially available from suppliers, such as Perkin Elmer / Applied BioSystems Division (Foster City, Calif.), and can be operated according to the manufacturer's instructions.

[0167] In certain embodiments, the nucleic acid encoding the antigen is DNA. This type of exemplary DNA-based vaccine contains DNA encoding one or more polypeptide antigens, so that the antigen is generated in situ. Alternatively, the vaccine may be an RNA-based vaccine. In certain embodiments, the nucleic acid encoding the antigen is mRNA. The mRNA may encode any polypeptide antigen of interest, including any naturally occurring or non-naturally occurring or otherwise modified polypeptide. The polypeptide encoded by the mRNA may be of any size and may have any secondary structure or activity. In some embodiments, the polypeptide encoded by the mRNA may stimulate an immune response when expressed in a cell.

[0168] The vaccine compositions of the present disclosure may also contain other compounds, which may be biologically active or inactive. The vaccine or medicament may include an adjuvant or immunostimulant, or a polynucleotide encoding an adjuvant or immunostimulant (e.g., an adjuvant polypeptide). Adjuvants and immunostimulants are compounds or compositions that directly or indirectly stimulate the immune system's response to a co-administered antigen. In some embodiments, the vaccine is unadjuvanted or self-adjuvanted.

[0169] Suitable adjuvants are commercially available, for example, as glucopyranosyl lipid adjuvant (GLA); Pam3CSK4; Freund's incomplete and complete adjuvant (Difco Laboratories, Detroit, Mich.); Merck adjuvant 65 (Merck and Company, Inc., Rahway, NJ); AS-2 (SmithKline Beecham; mineral salts (e.g., aluminum, silica, kaolin, and carbon); aluminum salts (e.g., aluminum hydroxide gel (alum), AlK(SO4)2, AlNa(SO4)2, AlNH4(SO4), and Al(OH)3); calcium salts (e.g., Ca3(PO4)2); iron or zinc; insoluble suspensions of acylated tyrosines; acylated sugars; cationic or anionic derivatized polysaccharides; polynucleotides (e.g., poly IC, poly AU acid, and CpG oligodeoxynucleotides (e.g., class A or B); polyphosphazenes; cyanoacrylates; polymerase-(DL-lactide-co-glycoside); bovine serum albumin; diphtheria toxoid; tetanus toxoid; edestin; keyhole limpet hemocyanin; Pseudomonas aeruginosa toxin A; choleragenoids; cholera toxin; pertussis toxin; viral proteins; Quil A, aminoalkyl glucosamine phosphate compounds. Additionally, adjuvants such as cytokines (e.g., GM-CSF or interleukin-2, interleukin-7, or interleukin-12), interferons, or tumor necrosis factors may also be used as adjuvants. Protein and polypeptide adjuvants may be obtained from natural or recombinant sources according to methods well known to those of skill in the art. If obtained from recombinant sources, the adjuvant may include a protein fragment that contains at least the immunostimulatory portion of the molecule.

[0170] Other known immunostimulatory polymers that can be used include, but are not limited to, polysaccharides, tRNA, non-metabolizable synthetic polymers (e.g., polyvinylamine, polymethacrylic acid, polyvinylpyrrolidone), mixed (relatively high molecular weight) polycondensates of 4',4-diaminodiphenylmethane-3,3'-dicarboxylic acid and 4-nitro-2-aminobenzoic acid (see Sela, M., Science 166:1365-1374 (1969)), or glycolipids, lipids, or carbohydrates.

[0171] In some embodiments, the adjuvant polypeptide comprises an immune activating protein (e.g., CD70, CD40 ligand, and constitutively active TLR4) or a polycationic peptide (e.g., protamine). In some embodiments, the adjuvant polypeptide is a flagellin polypeptide. Commercially available mRNA encoding the adjuvant polypeptide is available, for example, as TriMix (see Bonehill, A. et al. Mol. Ther. 16, 1170-1180 (2008), incorporated herein by reference). In some embodiments, the vaccine may comprise at least two separate polynucleotides, one encoding the anti-Mullerian hormone receptor II extracellular domain (AMHR2-ED) as described above, and the other encoding the adjuvant polypeptide (e.g., a flagellin polypeptide or an immune activating protein).

[0172] Vaccine preparation is a highly developed art, and general guidelines on vaccine preparation and formulation are readily available from any of a variety of sources. One such example is New Trends and Developments in Vaccines, edited by Volier et al. University Park Press, Baltimore, Md., USA 1978. Vaccine compositions may generally be used for prophylactic and therapeutic purposes.

[0173] The amount of antigen in each vaccine administration is generally selected as an amount that will induce an immunoprotective response without significant side effects in a typical vaccine. Such an amount will vary depending on which particular immunogen is used and how it is presented. Of course, the dosage administered may vary with age, weight, type of concurrent treatment, if any, and the nature of the antigen administered.

[0174] The immunogenic activity of a given amount of the vaccine composition can be readily determined, for example, by monitoring the increase in antibody titers against the antigen used in the vaccine composition (Dalsgaard, K. Acta Veterinia Scandinavica 69:1-40 (1978)). Another common method involves intradermal injection of various amounts of the vaccine composition into CD-1 mice, after which serum is collected from the mice and tested for anti-immunogen antibodies, for example, by ELISA. These and other similar approaches will be apparent to those skilled in the art.

[0175] 7.How to use The present disclosure provides a method for delivering an active agent to a cell. The method includes contacting the cell with a composition disclosed herein. In some embodiments, the cell is in vitro. In some embodiments, the cell is in vivo. Thus, the present disclosure further provides a method for delivering an active agent to a subject. The method may include administering to the subject a composition described herein.

[0176] The present disclosure also provides a method of producing a polypeptide of interest in a cell. The method includes contacting a cell with a composition disclosed herein, the composition includes an mRNA encoding the polypeptide of interest. In some embodiments, the cell is in vitro. In some embodiments, the cell is in vivo. Thus, the present disclosure provides a method of producing a polypeptide of interest in a subject, the method includes administering a composition described herein to a subject, the composition includes an mRNA encoding the polypeptide of interest.

[0177] The present disclosure further provides a method for treating a disease or disorder comprising administering a composition disclosed herein to a subject in need thereof. In some embodiments, the subject is a human.

[0178] The disease or disorder may include cancer, autoimmune diseases, inflammatory diseases, and infectious diseases.

[0179] In some embodiments, the disease or disorder is an inflammatory disease or disorder. Inflammatory diseases are characterized by activation of the immune system of a tissue or organ to abnormal levels that can cause abnormal function and / or disease of the tissue or organ. Inflammatory diseases and disorders that can be treated by the methods of the present invention include, but are not limited to, arthritis, rheumatoid arthritis, asthma, inflammatory bowel disease (Crohn's disease or ulcerative colitis), chronic obstructive pulmonary disease (COPD), allergic rhinitis, vasculitis (polyarteritis nodosa, temporal arteritis, Wegener's granulomatosis, Takayasu's arteritis, or Behcet's syndrome), inflammatory neuropathy, psoriasis, systemic lupus erythematosus (SLE), chronic thyroiditis, Hashimoto's thyroiditis, Addison's disease, polymyalgia rheumatica, Sjogren's syndrome, or Churg-Strauss syndrome.

[0180] In some embodiments, the disease or disorder is an autoimmune disease or disorder. Autoimmune diseases and disorders refer to a condition in a subject characterized by cell, tissue, and / or organ damage caused by the subject's immune response to its own cells, tissues, and / or organs. Autoimmune diseases and disorders that may be treated by the methods of the present invention include, but are not limited to, alopecia areata, ankylosing spondylitis, antiphospholipid syndrome, autoimmune Addison's disease, autoimmune disease of the adrenal gland, autoimmune hemolytic anemia, autoimmune hepatitis, autoimmune oophoritis and orchitis, autoimmune thrombocytopenia, Behcet's disease, bullous pemphigoid, cardiomyopathy, celiac sprue dermatitis, chronic fatigue immune deficiency syndrome (CFIDS), chronic inflammatory demyelinating polyneuropathy, Churg-Strauss syndrome, cicatricial pemphigoid, CREST syndrome, cold agglutinin disease, Crohn's disease, discoid lupus, mixed essential cryoglobulinemia, fibromyalgia-fibromyositis, glomerulonephritis, Graves' disease, Guillain-Barre syndrome, Hashimoto's thyroiditis, idiopathic pulmonary fibrosis, idiopathic thrombocytopenic purpura. Irritable bowel disease (ITP), Irritable bowel disease (IBD), IgA neuropathy, juvenile arthritis, lichen planus, lupus erythematosus, Meniere's disease, mixed connective tissue disease, multiple sclerosis, type 1 or immune-mediated diabetes mellitus, myasthenia gravis, pemphigus vulgaris, pernicious anemia, polyarteritis nodosa, polychondritis, polyglandular syndrome, polymyalgia rheumatica, polymyositis and dermatomyositis, primary agammaglobulinemia, Primary biliary cirrhosis, psoriasis, psoriatic arthritis, Raynaud's phenomenon, Reiter's syndrome, rheumatoid arthritis, sarcoidosis, scleroderma, Sjogren's syndrome, stiff man syndrome, systemic lupus erythematosus, lupus erythematosus, Takayasu's arteritis, temporal arteritis / giant cell arteritis, ulcerative colitis, uveitis, vasculitis (e.g., dermatitis herpetiformis, vasculitis), vitiligo, and Wegener's granulomatosis.

[0181] Some autoimmune disorders are also associated with inflammatory conditions.Examples of inflammatory disorders that are also autoimmune disorders that can be prevented, treated or managed according to the method of the present invention include, but are not limited to, asthma, encephalitis, inflammatory bowel disease, chronic obstructive pulmonary disease (COPD), allergy disorders, pulmonary fibrosis, undifferentiated spondyloarthropathy, undifferentiated arthropathy, arthritis, inflammatory osteolysis, and chronic inflammation caused by chronic viral or bacterial infection.Examples of types of psoriasis that can be treated according to the composition and method of the present invention include, but are not limited to, psoriasis vulgaris, pustular psoriasis, erythrodermic psoriasis, guttate psoriasis, and inverse psoriasis.

[0182] In some embodiments, the disease or disorder is cancer. In some embodiments, the cancer comprises a solid tumor. In some embodiments, the cancer comprises a hematological cancer or lymphoma. In some embodiments, the cancer is a metastatic cancer. In some embodiments, the disclosed compounds, compositions, or methods result in the inhibition of removal of metastases. In some embodiments, the disclosed compounds, compositions, or methods result in a reduction in tumor growth. In some embodiments, the disclosed compounds, compositions, or methods prevent tumor recurrence.

[0183] The compounds and compositions herein may be useful to treat a wide variety of cancers (e.g., carcinoma, sarcoma, lymphoma, leukemia, melanoma, mesothelioma, multiple myeloma, or seminoma). The cancer may be of the bladder, blood, bone, brain, breast, cervix, colon / rectum, endometrium, head and neck, kidney, liver, lung, lymph node, muscle tissue, ovary, pancreas, prostate, skin, spleen, stomach, testis, thyroid, or uterus.

[0184] In some embodiments, the cancer is an invasive and / or metastatic cancer (e.g., stage II cancer, stage III cancer, or stage IV cancer). In some embodiments, the cancer is an early stage cancer (e.g., stage 0 cancer, stage I cancer) and / or is not an invasive and / or metastatic cancer.

[0185] In some embodiments, the disease or disorder is an infectious disease. Infectious diseases that can be treated or prevented by the methods of the present invention are caused by infectious agents (such as, but not limited to, viruses, bacteria, fungi, protozoa, helminths, and parasites). The present invention is not limited to treating or preventing infectious diseases caused by intracellular or extracellular pathogens. Infectious diseases can be derived from bacteria (e.g., Mycobacterium tuberculosis, Chlamydia, Francisella tularensis), DNA viruses (e.g., Herpesviridae (Herpes simplex virus type 1, Kaposi's sarcoma-associated virus, and Epstein-Barr virus), Papillomaviridae (human papillomavirus), adenovirus, and Hepadnaviridae (Hepatitis B virus)), or RNA viruses (e.g., Retroviridae (human immunodeficiency virus), Flaviviridae (dengue virus, hepatitis C virus), Orthomyxoviridae (influenza), and Coronaviridae (human coronavirus and SARS coronavirus).

[0186] The compositions disclosed herein can be administered to a subject in a variety of ways. In any of the uses or methods described herein, administration can be by a variety of routes known to those skilled in the art, including, but not limited to, oral, inhaled, intravenous, intramuscular, topical, subcutaneous, systemic, and / or intraperitoneal administration to a subject in need thereof.

[0187] The amount of the disclosed composition required for use in the methods described herein will vary depending on the particular compound selected, as well as the route of administration, the nature and / or symptoms of the disease, and the age and condition of the patient, and will ultimately be left to the discretion of the attending physician or clinician. The determination of effective dosage levels (dosage levels required to achieve the desired results) can be performed by those skilled in the art using routine methods (e.g., human clinical trials, in vivo studies, and in vitro studies). For example, effective dosages can be determined by comparing in vitro activity and in vivo activity in animal models.

[0188] Dosage and dosing intervals can be individually adjusted to provide plasma levels of the active moiety sufficient to maintain the modulating effect, or minimum effective concentration (MEC). The MEC will vary from compound to compound, but can be estimated from in vivo and / or in vitro data. The dosage required to achieve the MEC will be determined by individual characteristics and route of administration. However, FIPLC assays or bioassays can be used to determine plasma concentrations. Dosing intervals can also be determined using the MEC value. The composition should be administered using a regimen that maintains plasma levels above the MEC 10-90% of the time, preferably 30-90%, most preferably 50-90%. In the case of local administration or selective uptake, the effective local concentration of the drug may not be related to the plasma concentration.

[0189] It should be noted that the attending physician would know how and when to terminate, interrupt, or adjust administration due to toxicity or organ dysfunction. Conversely, the attending physician would also know to adjust treatment to higher levels if the clinical response is not adequate (precluding toxicity). The magnitude of the administered dose in the management of the disorder of interest will vary with the severity of the condition to be treated and the route of administration. Furthermore, the dose, and perhaps the frequency of administration, will also vary with the age, weight, and response of the individual patient. Programs comparable to those discussed above may also be used in veterinary medicine.

[0190] The compositions disclosed herein can be evaluated for efficacy and toxicity using known methods. For example, the toxicity of a particular compound, or a subset of compounds sharing a particular chemical moiety, or compositions thereof, can be established by determining in vitro toxicity on a cell line (e.g., a mammalian cell line, preferably a human cell line). The results of such studies are often predictive of toxicity to animals (e.g., mammals), and more specifically, humans. Alternatively, the toxicity of a particular compound in an animal model (e.g., mice, rats, rabbits, dogs, or monkeys) can be determined using known methods. Efficacy can be established using several recognized methods (e.g., in vitro methods, animal models, or human clinical trials). When selecting a model to determine efficacy, one skilled in the art can be guided by the state of the art to select the appropriate model, dose, route of administration, and / or regimen.

[0191] A wide range of second therapies can be used in combination with the compounds and methods of the present disclosure. Second therapies can be administration of additional active agents or second therapies that are not associated with administration of another therapeutic agent. Such second therapies include, but are not limited to, surgery, immunotherapy, and radiation therapy.

[0192] The second therapy may be administered simultaneously with the first therapy, in the same composition, or in a separate composition that is administered substantially simultaneously with the first composition. In some embodiments, the second therapy may precede or follow the treatment of the first therapy by a time interval ranging from a few hours to a few months.

[0193] In some embodiments, the at least one additional active agent comprises an immunomodulatory agent, a chemotherapeutic agent, a nucleic acid (e.g., an mRNA, an aptamer, an antisense oligonucleotide, a ribozyme nucleic acid, an interfering RNA, an antigenic nucleic acid), a steroid, an analgesic agent, and an antimicrobial agent, or a combination thereof.

[0194] Exemplary immunomodulatory agents include: signal transducer and activator of transcription 3 (Stat3) inhibitors and analogs thereof (e.g., SM-36 and analogs thereof); Toll-like receptor (TLR) agonists and analogs thereof (e.g., imiquimod, resiquimod, sergantolimod, gardiquimod, SM-360320, TMX-101, TMX-202, TMX-302, TMX-306, GSK2245035, CL097, 852A, AZD-8848, DSP-3025, GS-9620, RO7020531, RO6871765, ANA773, DSP-0509, NJH395, BNT411, TQ-A3334, JNJ-4964, LHC165, CV8102, VTX-1463, VTX-2337, IMO-8400, IMO-3100, IRS-954, and their analogs); and statins or other lipid-lowering drugs and their analogs (e.g., atorvastatin, pravastatin, fluvastatin, simvastatin, lovastatin, mevastatin, pitavastatin, rosuvastatin, and their analogs).

[0195] In some embodiments, the additional active agent comprises at least one chemotherapeutic agent. As used herein, the term "chemotherapeutic agent" or "anticancer agent" includes any small molecule or other agent used in the treatment or prevention of cancer. Chemotherapeutic agents include, but are not limited to, cyclophosphamide, methotrexate, 5-fluorouracil, doxorubicin, docetaxel, daunorubicin, bleomycin, vinblastine, dacarbazine, cisplatin, paclitaxel, raloxifene hydrochloride, tamoxifen citrate, abemacirb, afinitor (everolimus), alpelisib, anastrozole, pamidronate, anastrozole, exemestane, capecitabine, epirubicin hydrochloride, eribulin mesylate, toremifene, fulvestrant, letrozole, gemcitabine, goserelin, ixabepilone, emtansine, lapatinib, olaparib, megestrol, neratinib, palbociclib, ribociclib, talazoparib, thiotepa, toremifene, methotrexate, and tucatinib. In selected embodiments, the chemotherapy agent comprises paclitaxel.

[0196] In some embodiments, the compositions can be administered in combination with an antimicrobial agent (eg, an antiviral or antibacterial agent). In some embodiments, the antibacterial agent is an antiviral agent (e.g., abacavir, acyclovir, adefovir, amantadine, amprenavir, atazanavir, baloxavir marboxil, bictarvy, boceprevir, brevirtide, cidofovir, cobicistat, daclatasvir, darunavir, delavirdine, didanosine, docosanol, dolutegravir, doravirine, edoxudine, efavirenz, elvitegravir, emtricitabine, enfuvirtide, entecavir, etravirine, famciclovir, fomivirsen, fosamprenavir, foscarnet, ganciclovir, ibacitabine, ibalizumab, idoxuridine, imiquimod, imunovir, indinavir, lamivudine, letermovir, lopinavir, loviride, maraviroc, mevir, serovar ... tisane, moroxydine, nelfinavir, nevirapine, nexavir, nitazoxanide, oseltamivir, penciclovir, peramivir, penciclovir, peramivir, pleconaril, podophyllotoxin, raltegravir, remdesivir, ribavirin, rilpivirine, rimantadine, lintatolimod, ritonavir, saquinavir, simeprevir, sofosbuvir, stavudine, taribavirin, telaprevir, telbivudine, tenofovir (e.g., tenofovir alafenamide or tenofovir disoproxil), tipranavir, trifluridine, tromantadine, umifenovir, valacyclovir, valganciclovir, vicriviroc, vidarabine, zalcitabine, zanamivir, and zidovudine, and combinations thereof.

[0197] In some embodiments, the antibacterial agent is an antimicrobial agent. Examples of antibacterial agents include sulfonamides, amphenicols, spectinomycin, trimethoprim, glycylcyclines, macrolides (e.g., erythromycin, clarithromycin, azithromycin, roxithromycin), oxazolidinones (e.g., linezolid), tetracyclines (e.g., doxycycline, tetracycline, minocycline), β-lactams (e.g., penicillin, methicillin, cloxacillin), carbapenems (e.g., imipenem, meropenem, aztreonam), aminoglycosides (e.g., gentamicin, tobramycin, amikacin), quinolones and fluoroquinolones (e.g., levofloxacin, ciprofloxacin, moxifloxacin), glycopeptides (e.g., vancomycin), and polymyxins (e.g., polymyxin, colistin).

[0198] In some embodiments, the second therapy comprises immunotherapy, which comprises chimeric antigen receptor (CAR) T cell therapy or T cell transfer therapy, cytokine therapy, immunomodulatory drugs, cancer vaccines, or administration of antibodies (e.g., monoclonal antibodies).

[0199] In some embodiments, immunotherapy involves the administration of antibodies. The antibodies may target either antigens specifically expressed by tumor cells or antigens shared with normal cells. In some embodiments, immunotherapy may include antibodies targeting, for example, CD20, CD33, CD52, CD30, HER (also called erbB or EGFR), VEGF, CTLA-4 (also called CD152), epithelial cell adhesion molecule (EpCAM, also called CD326), and PD-1 / PD-L1. Suitable antibodies include, but are not limited to, rituximab, blinatumomab, trastuzumab, gemtuzumab, alemtuzumab, ibritumomab, tositumomab, bevacizumab, cetuximab, panitumumab, ofatumumab, ipilimumab, brentuximab, pertuzumab, and the like. In some embodiments, the additional therapeutic agent may include an anti-PD-1 / PD-L1 antibody, including, but not limited to, pembrolizumab, nivolumab, cemiplimab, atezolizumab, avelumab, durvalumab, and ipilimumab. The antibody may also be conjugated to a chemotherapeutic agent. Thus, in some embodiments, the antibody is an antibody drug conjugate.

[0200] Immunotherapy (e.g., administration of an antibody) may be administered to a subject in a variety of ways. In any of the uses or methods described herein, administration may be by a variety of routes known to those skilled in the art (e.g., but not limited to, oral, inhaled, intravenous, intramuscular, topical, subcutaneous, systemic, and / or intraperitoneal administration to a subject in need thereof). Immunotherapy may be administered parenterally (e.g., but not limited to, subcutaneous, intramuscular, intravenous, intraperitoneal, intracardiac, and intraarticular injection). In some embodiments, immunotherapy may be administered in the same manner or in a different manner than the disclosed compounds or compositions.

[0201] 8. Kit In another aspect, the present disclosure provides a kit comprising at least one of the disclosed compounds or a pharma- ceutically acceptable salt thereof, or a composition comprising the compound or a pharma- ceutically acceptable salt thereof, and instructions for using the compound or composition. In some embodiments, the kit comprises at least one of the disclosed compounds, and one or more cationic and / or ionizable lipids, phospholipids, neutral or non-cationic lipids, polyethylene glycol (PEG) lipid conjugates, and / or structured lipids. The kit may further comprise at least one active agent. The kit may also comprise other agents and / or products that are co-packaged, co-formulated, and / or co-delivered with other components.

[0202] The kit may also include instructions for use of the components of the kit. Instructions are materials or methodologies related to the kit. The materials may include any combination of background information, list of components, simple or detailed protocols for using the compositions, troubleshooting, reference materials, technical support, and other related documentation. The instructions may be provided with the kit or as a separate member component, in paper or electronic form, provided on a computer readable memory device, downloaded from an internet website, or provided as a recorded presentation.

[0203] It is understood that the disclosed kits can be used in connection with the disclosed methods. The kits may further include containers or devices for use in the methods or compositions disclosed herein. The kits may optionally provide additional components, such as buffers and disposable, single-use equipment, such as pipettes, cell culture plates, or flasks.

[0204] The kits provided herein are in suitable packaging. Suitable packaging includes, but is not limited to, vials, bottles, jars, flexible packaging, etc. Individual member components of the kit may be physically packaged together or separately. EXAMPLES

[0205] 9. Working Example Abbreviations used in the schemes and examples below are as follows: BOC is tert-butyloxycarbonyl; DMAP is 4-dimethylaminopyridine; DMF is dimethylformamide; EDC is 1-ethyl-3-(3-dimethylaminopropyl)carbodiimide; eq is equivalents; EtOAc is ethyl acetate; MeOH is methanol; RT or rt is room temperature; THF is tetrahydrofuran.

[0206] All air- and moisture-sensitive manipulations were carried out under argon or in vacuum using standard Schlenk techniques. Anhydrous solvents (Et2O, THF, dioxane, DMSO, DMF, DCM, and toluene) were purchased from Fischer Scientific. All chemicals were purchased from Fischer Scientific, Sigma Aldrich, TCI, WUXI Apptec, and DC Chemicals Europe and used without further purification unless otherwise stated.

[0207] Analytical thin layer chromatography (TLC) was performed using Merck SIL G / UV254 plates. Compounds were visualized by exposure to UV light or by immersing the plates in a solution of ninhydrin or potassium permanganate followed by heating or staining with iodine vapor in a wide jar chamber. Column chromatography was performed in air with silica gel 60 (Fluka). Column chromatography was performed with Merck Kieselgel 60 (200-500 mm). Solvent systems were administered (s / sv:v).

[0208] NMR spectra 1H (300 MHz), 13C (75 MHz) were recorded on an ARX300 or Avance II 500 Bruker spectrometer, respectively. Chemical shifts (δ, ppm) are given relative to residual 1H or 13C of the deuterated solvents in the indicated solvents (CDCl3 7.26, 77.00; (CD3)2CO 2.05, 29.84, and 206.26, (CD3)2SO 2.50, 39.52). 1H-NMR and 13C-NMR chemical shifts (δ) are given in parts per million (ppm) relative to the TMS scale. Coupling constants J are given in Hz. The following abbreviations are used for the multiplicities in the proton spectra: s: singlet, d: doublet, t: triplet, q: quartet, qt: quintet, m: multiplet, br.: broad, dd: double doublet, dt: double triplet. Coupling constants (J) are reported in Hertz (Hz). Some signals that could not be assigned will be designated ArH (aromatic hydrogen).

[0209] Mass spectra (MS) were recorded on an LCQ-advantage (ThermoFinnigan) mass spectrometer equipped with positive (ESI+) or negative (ESI-) electrospray ionization (ionization voltage 4.5 kV, injection temperature 240°C).

[0210] Example 1 compound General Procedure 1: Esterification. Under nitrogen atmosphere, a solution of carboxylic acid derivative (1 eq.), EDC hydrochloride (1 eq.), and DMAP (0.5 eq.) in dry THF (0.1 M) was stirred at 0° C. for 0.5 h. Then, a solution of alcohol (1 eq.) in dry THF (0.1 M) was added. The solution was stirred at 0° C. to room temperature for 16 h. The progress of the reaction was monitored by TLC using a mixture of CH2-Cl2-MeOH (10:1) as eluent and 1H NMR using an ARX300 Brucker spectrometer. Upon completion of the reaction, the mixture was extracted with dichloromethane (3×). The combined organic layers were washed with 0.2 M aqueous HCl, saturated aqueous NaHCO3, and brine, respectively. The solid residue was then purified by manual column chromatography using CH2Cl2-EtOAc or CH2Cl2-MeOH stepwise gradient solvent systems as eluents to give the title product.

[0211] (9Z,12Z)-Octadeca-9,12-dien-1-yl 1-methyl-D-tryptophanate [ka] Step 1: Following general procedure 1, (9Z,12Z)-octadeca-9,12-dien-1-yl (9Z,12Z)-octadeca-9,12-dien-1-yl Na-(tert-butoxycarbonyl)-1-methyl-D-tryptophanate was dissolved in N α The compound was synthesized from 1-(tert-butoxycarbonyl)-1-methyl-D-tryptophan (1 equivalent), EDC hydrochloride (1 equivalent), DMAP (0.5 equivalent), and linoleyl alcohol (1 equivalent) at room temperature for 4 hours.

[0212] Step 2: (9Z,12Z)-Octadeca-9,12-dien-1-yl 1-methyl-D-tryptophanate was synthesized from (9Z,12Z)-octadeca-9,12-dien-1-yl Na-(tert-butoxycarbonyl)-1-methyl-D-tryptophanate (1 equiv.) by stirring in HCl solution in dioxane (HCl in dioxane (4M)) at 0 °C to RT for 5 h.

[0213] ((4-((1-methyl-D-tryptophyl)oxy)butyl)azanediyl)bis(hexane-6,1-diyl)bis(2-hexyldecanoate) [ka] Step 1: Following general procedure 1, ((4-((1-methyl-Na-pivaloyl-D-tryptophyl)oxy)butyl)azanediyl)bis(hexane-6,1-diyl)bis(2-hexyldecanoate) was dissolved in N α The compound was synthesized from 1-(tert-butoxycarbonyl)-1-methyl-D-tryptophan (1 equivalent), EDC hydrochloride (1 equivalent), DMAP (0.5 equivalent), and ALC-0315 (1 equivalent) at room temperature for 4 hours.

[0214] Step 2: ((4-((1-methyl-d-tryptophyl)oxy)butyl)azanediyl)bis(hexane-6,1-diyl)bis(2-hexyldecanoate) was synthesized from ((4-((1-methyl-Na-pivaloyl-d-tryptophyl)oxy)butyl)azanediyl)bis(hexane-6,1-diyl)bis(2-hexyldecanoate) (1 equiv.) by stirring in HCl solution in dioxane (HCl in dioxane (4 M)) from 0 °C to RT for 5 h. 1H NMR (500 MHz, CDCl3) δ 7.59 (d, J = 7.9 Hz, 1H), 7.28 (d, J = 8.2 Hz, 1H), 7.21 (t, J = 7.5 Hz, 1H), 7.09 (t, J = 7.4 Hz, 1H), 6.92 (s, 1H), 4.16 - 4.00 (m, 8H), 3.74 (s, 3H), 3.26 (dd, J = 14.4, 4.8 Hz, 1H), 3.01 (dd, J = 14.4, 7.8 Hz, 1H), 2.49 - 2.34 (m, 7H), 2.29 (td, J = 8.9, 4.5Hz, 3H), 1.67 - 1.52 (m, 15H), 1.32 - 1.17 (m, 52H), 0.86 (t, J = 6.8 Hz, 12H).

[0215] Heptadecan-9-yl 8-((4-((1-methyl-D-tryptophyl)oxy)butyl)(6-oxo-6-(undecyloxy)hexyl)amino)octanoate [ka] Step 1: Following general procedure 1, heptadecan-9-yl 8-((4-((1-methyl-Na-pivaloyl-D-tryptophyl)oxy)butyl)(6-oxo-6-(undecyloxy)hexyl)amino)octanoate was dissolved in N α The compound was synthesized from 1-(tert-butoxycarbonyl)-1-methyl-D-tryptophan (1 equivalent), EDC hydrochloride (1 equivalent), DMAP (0.5 eq), and SM-102 (1 equivalent) at room temperature for 4 hours.

[0216] Step 2: Heptadecan-9-yl 8-((4-((1-methyl-d-tryptophyl)oxy)butyl)(6-oxo-6-(undecyloxy)hexyl)amino)octanoate was synthesized from heptadecan-9-yl 8-((4-((1-methyl-Na-pivaloyl-d-tryptophyl)oxy)butyl)(6-oxo-6-(undecyloxy)hexyl)amino)octanoate (1 equiv.) by stirring in HCl solution in dioxane (HCl in dioxane (4 M)) at 0 °C to RT for 5 h. 1 H NMR (500 MHz, CDCl3) δ 7.58 (d, J = 7.9 Hz, 1H), 7.27 (d, J = 7.7 Hz, 1H), 7.20 (t, J = 7.5 Hz, 1H), 7.09 (t, J = 7.4 Hz, 1H), 6.92 (s, 1H), 4.89 - 4.80 (m, 1H), 4.15 - 4.08 (m, 2H), 4.06 - 4.01 (m, 2H), 3.80 - 3.75 (m, 1H), 3.74 (s, 3H), 3.25 (dd, J = 14.4, 4.8 Hz, 1H), 3.02 (dd, J = 14.4, 7.6 Hz, 1H), 2.62 (t, J = 6.4 Hz, 2H), 2.40 (dt, J = 8.0, 5.3 Hz, 4H), 2.26 (dt, J = 10.6, 7.5 Hz, 5H), 1.65 - 1.55 (m, 7H), 1.54 - 1.45 (m, 5H), 1.44 - 1.35 (m, 5H), 1.36 - 1.17 (m, 49H), 0.87 (t, J = 7.0 Hz, 9H).

[0217] Example 2 lipid composition Exemplary lipid compositions include any or all of: nucleic acids (e.g., mRNA, aptamers, antisense oligonucleotides, ribozyme nucleic acids, RNA interference, and antigen nucleic acids); modified lipids disclosed herein, phospholipids (e.g., phosphatidylcholine and phosphatidylethanolamine); cholesterol; and polyethylene glycol (PEG)-functionalized lipids (PEG lipids).

[0218] Cationic liposomes are generated by the dry film method. Lipids and cholesterol are dissolved in chloroform in a round-bottom flask. After removing the organic solvent by rotary evaporation at 40 °C, the dry lipid film is hydrated in PBS (pH 7.4) and vortexed six times for 30 s every 5 min at 45 °C. The resulting lipid suspension is subjected to 10 freeze-thaw cycles between liquid nitrogen and a 37 °C water bath and extruded with an extruder equipped with a 0.2 μm polycarbonate filter membrane to obtain a homogenous nanosuspension. Nucleic acid is then mixed with the liposomes and incubated at room temperature for 20 min to allow sufficient encapsulation.

[0219] The nucleic acid is prepared in acetate buffer at pH 4.0. Depending on the desired formulation, an ethanol solution containing the appropriate molar ratio of lipids is prepared.

[0220] The diameter, size distribution, polydispersity index (PDI), and zeta potential are measured with a ZETASIZER Nano (ZEN3600, Malvern Instruments, Malvern, Worcestershire, UK), and the morphology of lipid nanoparticles and liposomes is visualized using a JEOL2010F transmission electron microscope (TEM) and a FEI Quanta 200FEG scanning electron microscope (Phillips FEI Co., The Netherlands).

[0221] To detect the encapsulation efficiency (EE) and drug loading (DL), 40 μL of the final composition is taken and mixed with 160 μL of acetonitrile, followed by vortexing for 15 seconds and sonication in water for 5 minutes. After centrifugation at 12,000 rpm for 8 minutes, the supernatant is collected for high performance liquid chromatography (HPLC) measurement. EE and DL are calculated by the following formula: EE(%) = ma - mb / ma × 100% DL(%) = ma - mb / (ma+100mg)× 100% (wherein ma represents the amount of active drug (e.g., nucleic acid) in the initial chloroform solution, and mb represents the content of active drug in the lipid nanoparticles and liposomes detected by HPLC).

[0222] Example 3 In vitro and in vivo transfection, pharmacokinetics, and efficacy In vitro and in vivo transfection can be detected in cells or mice. To study the pharmacokinetics of the formulations, wide-type C57BL / 6 mice (male, 8 weeks old) were randomly assigned to different groups and administered the formulations 5 min, 0.5 h, 4 h, 8 h, 24 h, and 48 h after administration, blood was collected, and plasma was separated after centrifugation (13,300 rpm, 10 min). Subsequently, the mice were euthanized and major organs (e.g., liver, spleen, lungs, lymph nodes, fat pads, and tumors) were harvested, weighed, and homogenized. The content of the active drug in tissue homogenates and plasma was detected by liquid chromatography-mass spectrometry (LC-MS) using an ABI-5500 Qtrap (Sciex, Ontario, Canada) mass spectrometer equipped with an electrospray ionization source, coupled with a Shimadzu high-performance liquid chromatography (HPLC) system equipped with an Xbridge C18 column (50 × 2.1 mm ID, 3.5 μm). The efficacy of the formulation is evaluated in various disease models.

[0223] It is understood that the foregoing detailed description and accompanying examples are merely illustrative and do not limit the scope of the present disclosure, which is defined solely by the appended claims and equivalents thereof.

[0224] Various changes and modifications to the disclosed embodiments, which will be apparent to those skilled in the art, can be made without departing from the spirit and scope thereof.

Claims

1. Compound of formula (I) 【Chemistry 1】 or a pharmaceutically acceptable salt thereof (In the formula, R 1 is hydrogen, or C 1 -C 6 It is alkyl, X is O, NR 2 Selected from , S, and combination, R 2 is hydrogen, C 1 -C 6 -alkyl, C 2 -C 6 -alkenyl, C 2 -C 6 -alkynyl, C 3 -C 6 -cycloalkyl, or halo-C 1 -C 6 -alkyl, and L is C 8 -C 80 Alkyl, C 8 -C 80 Alkenil, C 8 -C 80 Alkinyl, C 8 -C 80 Heteroalkyl, C 8 -C 80 Heteroalkenyls, and C 8 -C 80 Selected from heteroalkynyls, each may be substituted with one or more substituents selected from hydroxyl and amino compounds.

2. R 1 The compound according to claim 1, or a pharmaceutically acceptable salt thereof, wherein the compound is methyl.

3. L is C 12 -C 40 Alkyl and C 12 -C 40 A compound selected from alkenyls, according to claim 1 or 2, or a pharmaceutically acceptable salt thereof.

4. L is a compound according to claim 1 or 2 having formula (A), or a pharmaceutically acceptable salt thereof: 【Chemistry 2】 (In the formula, n is 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, or 40. R a and R b Each of them is independent of C 6 -C 40 Alkyl, C 6 -C 40 Alkenil, C 6 -C 40 Heteroalkyl and C 6 -C 40 (Selected from heteroalkenyls).

5. The compound according to claim 4, or a pharmaceutically acceptable salt thereof, wherein n is 1, 2, 3, 4, 5, 6, 7, or 8.

6. L is a compound according to claim 1 or 2, having formula (D), (E), or (F), or a pharmaceutically acceptable salt thereof: 【Transformation 3】 (In the formula, n, p, and q are independently 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, or 40. R a1 and R a2 Each of them is independent of C 6 -C 40 Alkyl and C 6 -C 40 (Selected from Alkenil).

7. The compound according to claim 6, or a pharmaceutically acceptable salt thereof, wherein n, p, and q are each independently 1, 2, 3, 4, 5, 6, 7, or 8.

8. L is selected from the following: the compound according to claim 1 or 2, or a pharmaceutically acceptable salt thereof: 【Chemistry 4-1】 【Chemistry 4-2】 【Chemistry 4-3】

9. The compound according to claim 1, wherein the compound is selected from the group consisting of the following: 【Transformation 5】

10. A lipid composition comprising the compound described in claim 1.

11. The lipid composition according to claim 10, further comprising one or more of cationic and / or ionizable lipids, phospholipids, neutral or non-cationic lipids, structural lipids, polyethylene glycol (PEG)-lipid conjugates, or combinations thereof.

12. The lipid composition according to claim 11, wherein the structural lipid is selected from the group consisting of cholesterol, fecosterol, sitosterol, ergosterol, campesterol, stigmasterol, brassicasterol, tomatidine, cholesteryl hemisuccinate, cholesteryl sulfate, ursolic acid, alpha-tocopherol, and mixtures thereof.

13. The lipid composition according to claim 10 or 11, further comprising at least one active agent.

14. The lipid composition according to claim 13, wherein the at least one active agent is encapsulated in liposomes, lipid nanoparticles, or micelles in the lipid composition.

15. The lipid composition according to claim 13, wherein the at least one active agent comprises RNA.

16. A pharmaceutical composition comprising the lipid composition according to claim 10 or 11 and a pharmaceutically acceptable carrier.

17. A vaccine comprising the lipid composition according to claim 10 or 11 and an antigen or a nucleic acid encoding the same.

18. A lipid composition according to claim 13 for delivering an active drug to cells.

19. A lipid composition according to claim 10 or 11 for use in a method for generating a target polypeptide in cells, wherein the method comprises contacting the cells with the lipid composition, and the lipid composition comprises mRNA encoding the target polypeptide.

20. A pharmaceutical composition for treating or preventing a target disease or disorder, comprising the lipid composition according to claim 10 or 11, or a vaccine comprising the lipid composition according to claim 10 or 11 and an antigen or nucleic acid encoding the same.

21. A method for delivering an active drug to cells, comprising contacting the cells with the lipid composition described in claim 13, wherein the cells are in vitro.

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