Lipids, formulations and uses thereof

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 to 1-methyl-D-tryptophan) for the delivery of active agents and compositions and vaccines thereof.

Description

[0001] CROSS-REFERENCE TO RELATED APPLICATIONS

[0002] This application claims the benefit of U.S. Provisional Application No. 63 / 334,437, filed on April 25, 2022, the contents of which are incorporated herein by reference in their entirety. Technical Field

[0003] The present disclosure provides compounds, compositions, formulations, and methods for delivering active agents (eg, nucleic acids). Background Art

[0004] The ability to deliver macromolecules and active agents (e.g., nucleic acids) into cells is of great significance for therapeutic agents, diagnostic agents, reagents, and for bioassays. Recently, the delivery of therapeutic agents based on siRNA and mRNA has now been used clinically for coronavirus disease 2019 (COVID-19) and cancer. However, due to instability, low permeability, and unnecessary activation of innate immune responses, the delivery of nucleic acids is very difficult. A safe, effective, and stable delivery system is needed to facilitate the in vivo delivery of nucleic acids.

[0005] Lipid nanoparticles (LNP) and cationic liposomes have been shown to be effective nucleic acid delivery systems. The compositions of lipid nanoparticles or cationic liposomes include cationic lipids or ionizable lipids and other types of lipids, wherein the cations or ionizable lipids are functionalized to encapsulate nucleic acids and induce effective transfection in vivo. Although multiple such lipid nanoparticle compositions have been demonstrated, safety, efficacy and specificity aspects are still lacking in improvement. Summary of the invention

[0006] In one aspect, disclosed herein is a compound of formula (I):

[0007]

[0008] or a pharmaceutically acceptable salt thereof, wherein:

[0009] R 1 is hydrogen or C1-C6 alkyl;

[0010] X is selected from O, NR 2 , S and key;

[0011] R 2 is hydrogen, C1-C6 alkyl, C2-C6 alkenyl, C2-C6 alkynyl, C3-C6 cycloalkyl or halo-C1-C6 alkyl; and

[0012] L is selected from C8-C 80 Alkyl, C8-C 80 Alkenyl, C8-C 80 Alkynyl, C8-C80 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.

[0013] In some embodiments, R 1 It is methyl

[0014] In some embodiments, L is selected from C 12 -C 40 Alkyl and C 12 -C 40 In some embodiments, L has formula (A):

[0015]

[0016] in:

[0017] 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; and

[0018] R a and R b Each independently selected from C6-C 40 Alkyl, C6-C 40 Alkenyl, C6-C 40 Heteroalkyl and C6-C 40 Heteroalkenyl.

[0019] In some embodiments, n is 1, 2, 3, 4, 5, 6, 7, or 8.

[0020] In some embodiments, L is of formula (D), (E), or (F):

[0021]

[0022] in:

[0023] 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; and

[0024] R a1 and R a2 Each independently selected from C6-C40 Alkyl and C6-C 40 Alkenyl.

[0025] In some embodiments, n, p, and q are each independently 1, 2, 3, 4, 5, 6, 7, or 8.

[0026] In some embodiments, L is selected from:

[0027]

[0028]

[0029]

[0030]

[0031] In some embodiments, the compound is selected from the group consisting of:

[0032]

[0033]

[0034] On the other hand, disclosed herein is a lipid composition comprising a compound disclosed herein (e.g., a compound of formula (I)) or a pharmaceutically acceptable salt thereof. In some embodiments, the lipid composition further comprises one or more cationic and / or ionizable lipids, phospholipids, neutral or non-cationic lipids, or a combination thereof.

[0035] In some embodiments, the lipid composition further comprises a structural lipid. In some embodiments, the structural lipid is selected from the group consisting of cholesterol, coprosterol, sitosterol, ergosterol, campesterol, stigmasterol, brassicasterol, tomatine, cholesterol hemisuccinate, cholesterol sulfate, ursolic acid, alpha-tocopherol and mixtures thereof.

[0036] In some embodiments, the lipid composition further comprises a polyethylene glycol (PEG)-lipid conjugate.

[0037] In some embodiments, the lipid composition also includes at least one activating agent. In some embodiments, at least one activating agent is wrapped by liposomes, lipid nanoparticles, micelles in the lipid composition. In some embodiments, at least one activating agent includes nucleic acid or protein. In some embodiments, nucleic acid encoding antigen or its functional fragment.

[0038] In some embodiments, at least one active agent comprises RNA. In some embodiments, the RNA is selected from the group consisting of small interfering RNA (siRNA), asymmetric interfering RNA (aiRNA), micro RNA (miRNA), Dicer substrate RNA (dsRNA), small hairpin RNA (shRNA), messenger RNA (mRNA), ribozymes, and mixtures thereof.

[0039] In some embodiments, the at least one active agent comprises an immunomodulator, a chemotherapeutic agent, a steroid, an analgesic, an antibacterial agent, or a combination thereof.

[0040] In another aspect, disclosed herein is a pharmaceutical composition comprising an effective amount of a compound disclosed herein (eg, a compound of formula (I)) or a pharmaceutically acceptable salt thereof, or a lipid composition disclosed herein, and a pharmaceutically acceptable carrier.

[0041] On the other hand, disclosed herein is a vaccine comprising an effective amount of: a compound disclosed herein (e.g., a compound of formula (I) or a pharmaceutically acceptable salt thereof), a lipid composition as described herein, or a pharmaceutical composition disclosed herein (e.g., a composition comprising a compound of formula (I) or a pharmaceutically acceptable salt thereof, or a lipid composition as described herein); and an antigen or a nucleic acid encoding the same.

[0042] In some embodiments, the antigen is a tumor antigen, an autoantigen, or an infectious disease derived antigen. In some embodiments, the nucleic acid is messenger RNA (mRNA).

[0043] In another aspect, disclosed herein is a method for delivering an active agent to a cell, 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 pharmaceutically acceptable salt thereof, or a lipid composition as described herein), or a vaccine disclosed herein (e.g., a vaccine comprising a compound of formula (I) or a pharmaceutically acceptable salt thereof, or a lipid composition as described herein).

[0044] In another aspect, disclosed herein is a method for delivering an active agent to a subject, comprising administering to the subject a lipid composition disclosed herein, or a pharmaceutical composition disclosed herein (e.g., a pharmaceutical composition comprising a compound of formula (I) or a pharmaceutically acceptable salt thereof, or a lipid composition as described herein), or a vaccine disclosed herein (e.g., a vaccine comprising a compound of formula (I) or a pharmaceutically acceptable salt thereof, or a lipid composition as described herein).

[0045] On the other hand, disclosed herein is a method for producing a polypeptide of interest in a cell, 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 pharmaceutically acceptable salt thereof, or a lipid composition as described herein), or a vaccine disclosed herein (e.g., a vaccine comprising a compound of formula (I) or a pharmaceutically acceptable salt thereof, or a lipid composition as described herein), wherein the lipid composition or pharmaceutical composition comprises an mRNA encoding the polypeptide of interest. In some embodiments, the cell is in a subject and the contacting comprises administering to the subject.

[0046] In another aspect, disclosed herein is a method for treating or preventing a disease or condition, 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 pharmaceutically 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 pharmaceutically acceptable salt thereof, or a lipid composition as described herein), or a vaccine disclosed herein (e.g., a vaccine comprising a compound of formula (I) or a pharmaceutically acceptable salt thereof, or a lipid composition as described herein).

[0047] In some embodiments, the disease or condition comprises cancer, autoimmune disease, inflammatory disease or infectious disease. In some embodiments, the disease or condition is cancer. In some embodiments, the subject suffers from cancer, has suffered from cancer, is susceptible to cancer or has a family history of cancer. In some embodiments, cancer comprises solid tumors or hematological cancers. In some embodiments, cancer is metastatic cancer. In some embodiments, the method inhibits or eliminates cancer metastasis, reduces tumor growth, prevents tumor recurrence or any combination thereof. In some embodiments, administration comprises initial immunization and at least one subsequent immunization.

[0048] In some embodiments, the subject is a human.

[0049] 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 immunomodulator, a chemotherapeutic agent, a nucleic acid, a steroid, an analgesic, an antibacterial agent, or a combination thereof.

[0050] Other aspects and embodiments of the present disclosure will be apparent from the following detailed description. DETAILED DESCRIPTION

[0051] Described herein are modified lipids for liposome and lipid nanoparticle delivery of active agents (e.g., miRNA) that are useful for effective encapsulation, improved delivery, and increased downstream efficacy (e.g., due to macrophage polarization). Modified lipids include cations or ionizable lipids conjugated to amino acids or IDO inhibitors (e.g., 1-methyl-D-tryptophan).

[0052] The section headings used in this section and throughout the disclosure herein are for organizational purposes only and are not intended to be limiting.

[0053] 1. Definition

[0054] As used herein, the terms "comprise," "including," "having," "has," "may," "contain," and variations thereof are intended to be open transitional phrases, terms, or words that do not exclude the possibility of additional actions or structures. Unless the context clearly dictates otherwise, the singular forms "a," "an," and "the" include plural referents. The present disclosure also contemplates other embodiments "comprising," "consisting of," and "consisting essentially of" the embodiments or elements set forth herein, whether or not explicitly stated.

[0055] For the recitation of numerical ranges herein, each intermediate value with the same degree of precision is expressly contemplated. For example, for the range of 6-9, the numbers 7 and 8 are contemplated in addition to 6 and 9, and for the range of 6.0-7.0, 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.

[0056] Unless otherwise defined herein, scientific and technical terms used in connection with the present disclosure shall have the meanings commonly understood by those of ordinary skill in the art. The meaning and scope of the terms shall be clear; however, if there is any potential ambiguity, the definitions provided herein shall take precedence over any dictionary or external definitions. In addition, unless the context requires otherwise, 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.

[0057] The term "contacting" as used herein means to bring into contact, to be in contact, or to make contact. The term "contacting" as used herein means a state or condition of touching or being in direct or partial proximity.

[0058] As used herein, "polynucleotide", "oligonucleotide" or "nucleic acid" means at least two nucleotides covalently linked together. A polynucleotide can be a DNA, a genome and cDNA, an RNA or a hybrid, wherein the polynucleotide can contain a combination of deoxyribonucleotides and ribonucleotides and a combination of bases, including uracil, adenine, thymine, cytosine, guanine, inosine, xanthine, hypoxanthine, isocytosine and isoguanine. Nucleic acids, whether DNA or RNA, can contain non-natural nucleotides, modified nucleotides and / or non-nucleotide building blocks (building blocks) (e.g., "nucleotide analogs") that can exhibit the same functions as natural nucleotides. Nucleic acids can be obtained by chemical synthesis methods or by recombinant methods. A polynucleotide can be single-stranded or double-stranded or can contain parts of double-stranded sequences and single-stranded sequences. The depiction of a single strand also defines the sequence of a complementary strand. Therefore, nucleic acids also encompass the complementary strands of the depicted single strands. Many variants of nucleic acids can be used for the same purpose as a given nucleic acid. Therefore, nucleic acids also encompass substantially identical nucleic acids and their complementary sequences.

[0059] A "peptide" or "polypeptide" is a linked sequence of two or more amino acids linked by peptide bonds. Peptides and polypeptides include proteins, such as binding proteins, receptors, and antibodies. The terms "polypeptide" and "protein" are used interchangeably herein.

[0060] As used herein, "nucleic acid", "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 pages 793-800 (Worth Pub. 1982)). The present technology contemplates any deoxyribonucleotide, ribonucleotide or peptide nucleic acid component and any chemical variant thereof, such as methylated, hydroxymethylated or glycosylated forms of these bases, etc. The polymer or oligomer may be heterogeneous or homogeneous in composition and may be isolated from a naturally occurring source or may be artificially or synthetically produced. In addition, 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 homoduplex, heteroduplex and hybrid states. In some embodiments, the nucleic acid or nucleic acid sequence comprises other types of nucleic acid structures, such as, for example, DNA / RNA helices, peptide nucleic acids (PNAs), morpholino nucleic acids (see, for example, Braasch and Corey, Biochemistry, 41(14):4503-4510 (2002)) and U.S. Pat. No. 5,034,506), locked nucleic acids (LNAs; see Wahlestedt et al., Proc. Natl. Acad. Sci. USA, 97:5633-5638 (2000)), cyclohexenyl nucleic acids (see Wang, J. Am. Chem. Soc., 122:8595-8602 (2000)) and / or ribozymes. Therefore, the term "nucleic acid" or "nucleic acid sequence" may also encompass chains comprising non-natural nucleotides, modified nucleotides, and / or non-nucleotide building blocks (e.g., "nucleotide analogs") that may exhibit the same functions as natural nucleotides; in addition, as used herein, the term "nucleic acid sequence" refers to oligonucleotides, nucleotides, or polynucleotides, 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 represent sense or antisense strands. The terms "nucleic acid," "polynucleotide," "nucleotide sequence," and "oligonucleotide" are used interchangeably. It refers to a polymeric form of nucleotides (deoxyribonucleotides or ribonucleotides, or analogs thereof) of any length.

[0061] As used herein, the terms "providing," "administering," and "introducing" are used interchangeably herein and refer to placing an active agent into a subject by a method or route that results in at least partial localization to a desired site.

[0062] A "subject" or "patient" can be human or non-human, and can include, for example, animal strains or species used as "model systems" for research purposes, such as the mouse model described herein. Likewise, patients can include adults or adolescents (e.g., children). In addition, a patient can refer to any living organism, preferably a mammal (e.g., human and non-human), that can benefit from the administration of the compositions contemplated herein. Examples of mammals include, but are not limited to, any member of the class mammals: humans, non-human primates such as chimpanzees and other apes and monkey species; farm animals such as cattle, horses, sheep, goats, pigs; domestic animals such as rabbits, dogs and cats; laboratory animals include rodents such as rats, mice and guinea pigs, etc. Examples of non-mammals include, but are not limited to, birds, fish, etc. In one embodiment, the mammal is a human.

[0063] As used herein, "treat," "treating," and the like means to slow, stop, or reverse the progression of a disease or condition when a compound or composition described herein is provided to an appropriate control subject. The term also means to reverse the progression of such a disease or condition to the extent that the symptoms are eliminated or greatly alleviated. Thus, "treating" refers to applying or administering a composition described herein to a subject, wherein the subject suffers from a disease or symptom of a disease, for the purpose of curing, healing, alleviating, alleviating, altering, remedying, improving, modifying, or affecting the disease or symptom of a disease.

[0064] The definitions of specific functional groups and chemical terms are described in more detail below. For the purposes of this disclosure, chemical elements are identified according to the CAS version of the periodic table, Handbook of Chemistry and Physics, 75th edition, inside cover, and specific functional groups are generally defined as described therein. In addition, the general principles of organic chemistry and specific functional parts and reactivity are described in the following: Sorrell, Organic Chemistry, 2nd edition, University Science Books, Sausalito, 2006; Smith, March's Advanced Organic Chemistry: Reactions, Mechanism, and Structure, 7th edition, John Wiley & Sons, Inc., New York, 2013; Larock, Comprehensive Organic Transformations, 3rd edition, John Wiley & Sons, Inc., New York, 2018; and Carruthers, Some Modern Methods of Organic Synthesis, 3rd edition, Cambridge University Press, Cambridge, 1987; The entire contents of each of them are incorporated herein by reference.

[0065] The term "alkyl" as used herein refers to a straight or branched saturated 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-dimethylpent-2-yl, n-heptyl, n-octyl, n-nonyl, n-decyl, undecyl, dodecyl, tridecyl, tetradecyl, pentadecyl, hexadecyl, heptadecyl, octadecyl, nonadecyl, and eicosyl.

[0066] 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 may be located at any position of the hydrocarbon chain. Representative examples of alkenyl include, but are not limited to, vinyl, 2-propenyl, 2-methyl-2-propenyl, 3-butenyl, 4-pentenyl, 5-hexenyl, 2-heptenyl, 2-methyl-1-heptenyl, and 3-decenyl.

[0067] As used herein, the term "alkynyl" means a straight or branched hydrocarbon chain containing at least one carbon-carbon triple bond. The triple bond may be located at any position of the hydrocarbon chain. Representative examples of alkynyl include, but are not limited to, ethynyl, propynyl, and butynyl.

[0068] The term "amino" as used herein refers to a -NH2 group. The term "alkylamino" as used herein refers to a group -NHR, wherein R is an alkyl group as defined herein. The term "dialkylamino" as used herein refers to a group -NR2, wherein each R is independently an alkyl group as defined herein.

[0069] As used herein, the term "cycloalkyl" refers to a saturated carbocyclic ring system containing three to ten carbon atoms and zero heteroatoms. The cycloalkyl can be monocyclic, bicyclic, bridged, fused or spirocyclic. Representative examples of cycloalkyl include, but are not limited to, cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, cycloheptyl, cyclooctyl, cyclononyl, cyclodecyl, adamantyl, bicyclo[2.2.1]heptyl, bicyclo[3.2.1]octyl and bicyclo[5.2.0]nonyl.

[0070] As used herein, the term "halogen" or "halo" means F, Cl, Br or I.

[0071] The term "haloalkyl" as used herein refers to an alkyl as defined herein, wherein at least one hydrogen atom (e.g., one, two, three, four, five, six, seven, or eight hydrogen atoms) is replaced by halogen. In some embodiments, each hydrogen atom of the alkyl is replaced by 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.

[0072] As used herein, the term "heteroalkyl" means an alkyl group, as defined herein, in which one or more carbon atoms (and any associated hydrogen atoms) are each independently replaced by a heteroatom group such as -NH-, -O-, -S-, -S(O)-, -S(O)2-, -OP(O)(O)2-, -NH-, -O-, -S-, -S(O)-, -S(O)2 ... - )O-, etc. For example, 1, 2, 3, 4, 5, 6 or more carbon atoms can be independently replaced by the same or different heteroatom groups. The heteroalkyl group can also contain one or more carbonyl moieties (i.e., wherein the carbon atoms of the alkyl group are oxidized to -C(O)- groups).

[0073] As used herein, the term "heteroalkenyl" refers to an alkenyl group as defined herein, wherein one or more carbon atoms (and any associated hydrogen atoms) are each independently replaced by a heteroatom group such as -NH-, -O-, -S-, -S(O)-, -S(O)2-, and the like. For example, 1, 2, 3, 4, 5, 6 or more carbon atoms may be independently replaced by the same or different heteroatom groups. The heteroalkenyl group may also contain one or more carbonyl moieties (i.e., wherein a carbon atom of an alkyl group is oxidized to a -C(O)- group).

[0074] As used herein, the term "heteroalkynyl" refers to an alkynyl group as defined herein, wherein one or more carbon atoms (and any associated hydrogen atoms) are each independently replaced by a heteroatom group such as -NH-, -O-, -S-, -S(O)-, -S(O)2-, and the like. For example, 1, 2, 3, 4, 5, 6 or more carbon atoms may be independently replaced by the same or different heteroatom groups. The heteroalkynyl group may also contain one or more carbonyl moieties (i.e., wherein a carbon atom of the alkyl group is oxidized to a -C(O)- group).

[0075] As used herein, the term "hydroxy" refers to an -OH group.

[0076] As used herein, the term "substituent" refers to a group that replaces an atom of the indicated group.

[0077] When 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 indicated in the expression using "substituted" can be replaced with a series of listed indicated groups or with suitable substituents known to those skilled in the art (e.g., one or more of the groups listed below), provided that the normal valence of the designated 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, hydrazine, imino, oxo, nitro, phosphate, phosphonate, sulfonic acid, sulfonamido, thiol, thiol, thioketone, thioloxy, or a combination thereof.

[0078] As used herein, in chemical structures, indicates:

[0079]

[0080] Indicates the point where one part joins another.

[0081] In some cases, the number of carbon atoms in a hydrocarbyl substituent (e.g., alkylalkenyl) is indicated by the prefix "C x -C y ” indicates an alkyl group containing from 1 to 3 carbon atoms, wherein x is the minimum number of carbon atoms in the substituent and y is the maximum number of carbon atoms. Thus, for example, “C1-C3 alkyl” refers to an alkyl substituent containing from 1 to 3 carbon atoms.

[0082] For the compounds described herein, radicals and substituents may be chosen with regard to the allowed valences of atoms and substituents such that selection and substitution results in stable compounds, eg, which do not spontaneously transform such as by rearrangement, cyclization, elimination, and the like.

[0083] Where substituent groups are designated by conventional chemical formulas written from left to right, they optionally encompass substituents resulting from writing the structure from right to left, e.g., -CH2O- is intended to encompass -OCH2-, and -C(O)NH- is intended to encompass -NHC(O)-.

[0084] Although methods and materials similar or equivalent to those described herein can be used in the practice or testing of the present disclosure, preferred methods and materials are described below. All publications, patent applications, patents and other references mentioned herein are incorporated by reference in their entirety. The materials, methods and embodiments disclosed herein are illustrative only and are not restrictive.

[0085] 2. Compounds

[0086] In one aspect, disclosed is a compound of formula (I):

[0087]

[0088] or a pharmaceutically acceptable salt thereof, wherein:

[0089] R 1 is hydrogen or C1-C6 alkyl;

[0090] X is selected from O, NR 2 , S and key;

[0091] R 2 is hydrogen, C1-C6 alkyl, C2-C6 alkenyl, C2-C6 alkynyl, C3-C6 cycloalkyl or halo-C1-C6 alkyl; and

[0092] L is selected from 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.

[0093] In some embodiments, R 1 is hydrogen. In some embodiments, R 1 is C1-C6 alkyl; in some embodiments, R 1 It is methyl.

[0094] 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 , where R2 is selected from hydrogen and C1-C6 alkyl. In some embodiments, X is NH. In some embodiments, X is NR 2 , where R 2 In some embodiments, X is S.

[0095] In some embodiments, L is a lipid moiety having at least 8 carbon atoms. The lipid moiety can be derived from any suitable lipid, such as a fatty alcohol, a fatty acid, a phospholipid, a steroid or a synthetic lipid. In some embodiments, the lipid moiety is derived from a lipid having a functional group (such as a hydroxyl, a carboxylic acid group or an amino group), and the lipid moiety is connected 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, if the lipid moiety is derived from a fatty alcohol, X is O.

[0096] In some embodiments, L is selected from 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.

[0097] For example, in some embodiments, L is selected from C8-C 80 Alkyl and C8-C 80 In some embodiments, L is selected from C8-C 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)-hexadeca-9-en-1-ol), oleyl alcohol ((Z)-octadeca-9-en-1-ol), anti-oleyl alcohol (trans-9-octadecaenol), cis-isooctadecaenol (cis-11-octadecaenol), gadol ((Z)-eicos-9-en-1-ol), 11-eicosene alcohol, erucyl alcohol (cis-13-eicosene alcohol), 15-eicosene-1-ol, eicosene alcohol (eicos-11,14-dien-1-ol), linolenic alcohol ((9Z, 12Z, 15Z)-9,1 2,15-octadecatriene-1-ol), γ-linolenic acid ((6E,9E,12E)-octadecatriene-6,9,12-triene-1-ol), tung oil alcohol (octadecatriene-9,11,13-triene-1-ol), eicosane-5,8,11-triene-1-ol, docosane-13-ene-1-ol, eicosane-11,14-17-triene-1-ol, octadecatriene-6,9,12,15-tetraene-1-ol, arachidonic acid ((5Z,8Z,11Z,14Z)-eicosatetraene-1-ol), 4E,6Z-hexadecadiene-1-ol, eicosane-5,8,11,14,17-pentaene-1-ol, docosahexaenol (docosahexaenol) ,7,10,13,16,19-hexaen-1-ol), 2-docosahexa-7,10,13,16,19-pentaen-1-ol, 2-tetradecanoic acid-6,9,12,15,18,21-hexaen-1-ol, n-octanol (1-octanol), nonanol (1-nonanol), decanol (1-decanol), undecanol (1-undecanol), lauryl alcohol (1-dodecanol), tridecanol (1-tridecanol), myristyl alcohol (1-tetradecanol), pentadecanol (1-pentadecanol), cetyl alcohol (1-hexadecanol), palmitoleyl alcohol (cis-9-hexadecene-1-ol), heptadecanol (1-n-heptadecanol), stearyl alcohol (1-octadecanol), oleyl alcohol (1-octadecenol), nonadecanol (1-nonadecanol), Arachidyl alcohol (1-eicosanol), heneicosanol (1-heneicosanol), behenyl alcohol (1-docosanol), erucyl alcohol (cis-13-docosan-1-ol), 1-tricosanol, tetracosanol (1-tetracosanol), pentacosanol (1-pentacosanol), hexacosanol (1-hexacosanol), 1-heptacosanol, montanol (1-octacosanol), 1-nonacosanol, myricanol (1-triacontanol), 1-triacontanol, 1-dotriacontanol (margarine alcohol), 1-triacontanol, candacetyl alcohol (1-tetratriacontanol), 1-hexatriacontanol, 1-heptatriacontanol, 1-octaticontanol, nonatriacontanol, 1-triacontanol, or 1-tetracontanol.

[0098] In some embodiments, L is selected from C8-C 80 Heteroalkyl, C8-C 80 Heteroalkenyl and C8-C 80 In such embodiments, L can be derived from a lipid comprising one or more heteroatom groups (such as -O-, -NH-, -C(O)-, etc.) or combinations thereof (e.g., a -C(O)O- group).

[0099] For example, in some embodiments, L has formula (A):

[0100]

[0101] in:

[0102] 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; and

[0103] R a and R b Each independently selected from 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 Each independently selected from C6-C 40 Alkyl and C6-C 40 For example, in some embodiments, R a and R bEach of the following is independently selected from the group consisting of n-hexyl, n-heptyl, n-octyl, n-nonyl, n-decyl, undecyl, dodecyl, tridecyl, tetradecyl, pentadecyl, hexadecyl, heptadecyl, octadecyl, nonadecyl, eicosyl, heneicosyl, docosyl, tricosyl, tetracosyl, pentacosyl, hexacosyl, heptacosyl, octacosyl, nonadecyl, triacontyl, triuncontyl, dotriacontyl, trictriacontyl, tetratriacontyl, pentatriacontyl, hexatriacontyl, heptatriacontyl, octatriacontyl, Nonadriatriadecyl, tetradecyl, linoleyl ((9Z,12Z)-octadec-9,12-dien-1-yl), palmityl ((Z)-hexadec-9-en-1-yl), oleyl ((Z)-octadec-9-en-1-yl), trans-oleyl (trans-9-octadecenyl), cis-11-octadecenyl (cis-vaccenyl) (cis-11-octadecenyl), gadyl ((Z)-eicos-9-en-1-yl), 11-eicosenyl, erucyl (cis-13-eicosenyl), 15 -Eicosyl, docosyl (eicosyl-11,14-dien-1-yl), linoleyl ((9Z,12Z,15Z)-9,12,15-octadecadien-1-yl), γ-linoleyl ((6E,9E,12E)-octadecadien-6,9,12-trien-1-yl), tung oil (octadecadien-9,11,13-trien-1-yl), eicosyl-5,8,11-trien-1-yl, docosyl-13-en-1-yl, eicosyl-11,14-17-trien-1-yl, In some embodiments, R a and R b Each is linoleyl.

[0106] In some embodiments, L is of formula (B) or (C):

[0107]

[0108] in:

[0109] 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;

[0110] R a1 Selected from C6-C 40 Alkyl and C6-C 40 alkenyl; and

[0111] R b Selected from C6-C 40 Alkyl, C6-C 40 Alkenyl, C6-C 40 Heteroalkyl and C6-C 40 Heteroalkenyl.

[0112] 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 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.

[0114] In some embodiments, R a1 C6-C 24 Alkyl or C6-C 24 In some embodiments, R a1 Selected from C9-C 22 Alkyl and C9-C 22 In some embodiments, R a1 Selected from straight or branched chain C 6、 C 7、 C 8、 C 9、 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、 C23、 or C 24 In some embodiments, R a1 Selected from straight or branched chain C 6、 C 7、 C 8、 C 9、 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 Selected from linoleyl, n-nonyl, n-undecyl, heneicosane-11-yl, pentadecane-7-yl and heptadecan-9-yl.

[0115] In some embodiments, R b Selected from C6-C 40 Alkyl and C6-C 40 For example, in some embodiments, R bis selected from the group consisting of n-hexyl, n-heptyl, n-octyl, n-nonyl, n-decyl, undecyl, dodecyl, tridecyl, tetradecyl, pentadecyl, hexadecyl, heptadecyl, octadecyl, nonadecyl, eicosyl, heneicosyl, docosyl, tricosyl, tetracosyl, pentacosyl, hexacosyl, heptacosyl, octacosyl, nonacosyl, triacontyl, triuncontyl, dotriacontyl, trictriacontyl, tetratriacontyl, pentatriacontyl, hexatriacontyl, heptatriacontyl, octatriacontyl, Nonatriacontyl, tetracontyl, linoleyl ((9Z, 12Z)-octadec-9,12-dien-1-yl), palmityl ((Z)-hexadec-9-en-1-yl), oleyl ((Z)-octadec-9-en-1-yl), transoleyl (trans-9-octadecenyl), cis-11-octadecenyl (cis-11-octadecenyl), gaddenyl ((Z)-eicos-9-en-1-yl), 11-eicosenyl, erucyl (cis-13-eicosenyl), 15-eicosene-1-yl, Docosyl (eicosyl-11,14-dien-1-yl), linoleyl ((9Z,12Z,15Z)-9,12,15-octadecatrien-1-yl), γ-linoleyl ((6E,9E,12E)-octadeca-6,9,12-trien-1-yl), tung oilyl (octadeca-9,11,13-trien-1-yl), eicosyl-5,8,11-trien-1-yl, eicosyl-13-en-1-yl, eicosyl-11,14-17-trien-1-yl, octadeca-6, In some embodiments, R b Linoleyl.

[0116] In some embodiments, L is of formula (D), (E), or (F):

[0117]

[0118] in:

[0119] 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; and

[0120] R a1 and R a2 Each independently selected from C6-C 40 Alkyl and C6-C 40 Alkenyl.

[0121] In some embodiments, L is of formula (D). In some embodiments, L is of formula (E). In some embodiments, L is of 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.

[0122] 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.

[0123] In some embodiments, R a1 and R a2 Each independently selected from C6-C 24 Alkyl and C6-C 24 In some embodiments, R a1 and R a2 Each independently selected from C9-C 22 Alkyl and C9-C 22 In some embodiments, R a1 and R a2 Each independently selected from a linear or branched C 6、 C 7、 C 8、 C 9、 C 10、 C 11、 C 12、 C 13、 C 14、 C 15、 C 16、 C 17、 C 18、 C 19、 C20、 C 21、 C 22、 C 23、 or C 24 In some embodiments, R a1 and R a2 Each independently selected from a linear or branched C 6、 C 7、 C 8、 C 9、 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 Each is independently selected from linoleyl, n-nonyl, n-undecyl, heneicos-11-yl, pentadec-7-yl and heptadecan-9-yl.

[0124] In some embodiments, L is derived from a steroid. For example, in some embodiments, L is derived from cholesterol, β-sitosterol, or BHEM-cholesterol.

[0125] In some embodiments, L is selected from:

[0126]

[0127]

[0128]

[0129]

[0130] Compounds can exist as stereoisomers, wherein there is an asymmetric or chiral center. A stereoisomer is "R" or "S", depending on the configuration of the substituents around the chiral carbon atom. The terms "R" and "S" used herein are configurations defined in IUPAC 1974 Recommendations for Section E, Fundamental Stereochemistry, Pure Appl. Chem., 1976, 45: 13-30. The present disclosure contemplates various stereoisomers and mixtures thereof, and these are specifically included within the scope of the present disclosure. Stereoisomers include enantiomers and diastereomers, as well as mixtures of enantiomers or diastereomers. A single stereoisomer of a compound can be synthesized from commercially available starting materials containing an asymmetric or chiral center, or by preparing a racemic mixture followed by a resolution method well known to those of ordinary skill in the art. Examples of these resolution methods are as follows: (1) attaching a mixture of enantiomers to a chiral auxiliary, separating the resulting diastereomeric mixture by recrystallization or chromatography, and optionally liberating the optically pure product 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 its most recent edition), or (2) directly separating the mixture of optical enantiomers on a chiral chromatographic column, or (3) fractional recrystallization methods.

[0131] It is to be understood that compounds may have tautomeric forms as well as geometric isomers and these also constitute embodiments of the present disclosure.

[0132] The present disclosure also includes isotopically labeled compounds, which are identical to those described in formula (I), but for the fact that one or more atoms are replaced by atoms having an atomic mass or mass number different from the atomic mass or mass number usually found in nature. Examples of isotopes include isotopes of hydrogen, carbon, nitrogen, oxygen, phosphorus, sulfur, fluorine and chlorine, such as, but not limited to, 2 H. 3 H. 13 C. 14 C. 15 N. 18 O. 17 O. 31 P. 32 P. 35 S. 18 F and 36Cl. With heavier isotopes (such as deuterium, e.g. 2 H) substitution may provide certain therapeutic advantages resulting from greater metabolic stability, such as increased in vivo half-life or reduced dosage requirements, and may therefore be preferred in certain circumstances. The compounds may incorporate positron emitting isotopes for use in medical imaging and positron emission tomography (PET) studies to determine receptor distribution. Suitable positron emitting isotopes that may be incorporated into the 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 an appropriate isotopically labeled reagent in place of a non-isotopically labeled reagent.

[0133] The disclosed compounds may exist as pharmaceutically acceptable salts. The term "pharmaceutically acceptable salt" refers to a salt or zwitterion of a compound that is water-soluble or oil-soluble or dispersible, suitable for treating a condition without excessive toxicity, irritation, and allergic reactions, commensurate with a reasonable benefit / risk ratio, and effective for its intended use. Salts can be prepared during the final separation and purification of the compound, or prepared separately by reacting the amino group of the compound with a suitable acid. For example, the compound can be dissolved in a suitable solvent, such as, but not limited to, methanol and water, and treated with at least one equivalent of an acid (such as hydrochloric acid). The resulting salt can be precipitated, separated by filtration, and dried under reduced pressure. Alternatively, the solvent and excess acid can be removed under reduced pressure to provide a salt. 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, naphthylsulfonate, nicotinate, oxalate, pamoate, pectinate, persulfate, 3-phenylpropionate, picrate, oxalate, maleate, pivalate, propionate, succinate, tartrate, trichloroacetate, trifluoroacetate, glutamate, p-toluenesulfonate, undecanoate, hydrochloride, hydrobromide, sulfate, phosphate, etc. The amino group of the compound can also be quaternized with alkyl chlorides, bromides and iodides such as methyl, ethyl, propyl, isopropyl, butyl, lauryl, myristyl, stearyl, etc.

[0134] Basic addition salts can be prepared by reacting the carboxyl group with a suitable base such as a hydroxide, carbonate or bicarbonate of a metal cation (such as lithium, sodium, potassium, calcium, magnesium or aluminum), or an organic primary, secondary or tertiary amine during the final isolation and purification of the disclosed compounds. Quaternary ammonium salts 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-diphenylhydroxymethylamine and N,N′-dibenzylethylenediamine, ethylenediamine, ethanolamine, diethanolamine, piperidine, piperazine, etc. can be prepared.

[0135] Compounds can be synthesized according to a variety of methods, including those shown in the examples. The reaction conditions and reaction times of each individual step can vary according to the specific reactants used and the substituents present in the reactants used. Specific procedures are provided in the Examples section. The reaction can be carried out in a conventional manner, for example, by removing the solvent from the residue, and further purified according to methods generally known in the art, such as but not limited to crystallization, distillation, extraction, wet grinding and chromatography. Unless otherwise described, starting materials and reagents are commercially available or can be prepared from commercially available materials by those skilled in the art using the methods described in the chemical literature. Starting materials, if not commercially available, can be prepared by a procedure selected from standard organic chemistry techniques, a technique similar to the synthesis of known structurally similar compounds, or a technique similar to the procedure described in the above-mentioned scheme or the Synthesis Example section.

[0136] Routine experiments, including appropriate manipulation of reaction conditions, reagents and synthetic route sequences, protection of any chemical functional groups incompatible with the reaction conditions, and deprotection at appropriate points in the reaction sequence of the method are all included in the scope of the present disclosure. Suitable protecting groups and methods of using such suitable protecting groups to protect different substituents and deprotection are well known to those skilled in the art; examples thereof can be found in PGM Wuts and TW Greene, in Greene's booktitled Protective Groups in Organic Synthesis (4th edition), John Wiley & Sons, NY (2006), which is incorporated herein by reference in its entirety. The synthesis of the compounds of the present disclosure can be accomplished by methods similar to those described in the synthetic schemes described above and the specific embodiments.

[0137] When an optically active form of a disclosed compound is required, it can be obtained by carrying out one of the procedures described herein using optically active starting materials (e.g., by asymmetric induced preparation by appropriate reaction steps) or by resolving the mixture of stereoisomers of the compounds or intermediates using standard procedures (such as chromatographic separation, recrystallization or enzymatic resolution).

[0138] Similarly, when pure geometric isomers of compounds are required, they may be obtained by carrying out one of the above procedures using the pure geometric isomers as starting materials, or by resolving a mixture of geometric isomers of compounds or intermediates using standard procedures such as chromatographic separation.

[0139] It should 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 included within the scope of the claims.

[0140] 3. Composition

[0141] The present disclosure also provides a lipid composition suitable for administration to a subject (such as a patient, which may be human or non-human), comprising a compound of formula (II):

[0142] WXL (II)

[0143] or a pharmaceutically acceptable salt thereof, wherein:

[0144] W is an amino acid;

[0145] X is selected from O, NR 2 , S and key;

[0146] R 2 is hydrogen, C1-C6 alkyl, C2-C6 alkenyl, C2-C6 alkynyl, C3-C6 cycloalkyl or halo-C1-C6 alkyl; and

[0147] L is selected from 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.

[0148] As used herein, the term "amino acid" 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 containing an amine and a carboxyl terminal functional group. In some embodiments, the term "amino acid" refers to all amino acid compounds that are similar to naturally occurring amino acids in structure and / or overall shape. Amino acids can be D-amino acids or L-amino acids. Natural amino acids include amino acids found in nature, such as amino acids that are combined into peptide chains to form a large number of protein building blocks (building-block). These are mainly L stereoisomers, although some D-amino acids are present in bacterial envelopes and some antibiotics. "Non-standard" natural amino acids include, for example, pyrrolysine (present in methanogenic organisms and other eukaryotic organisms), selenocysteine ​​(present in many non-eukaryotic organisms and most eukaryotic organisms) 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 not naturally encoded or found in the genetic code) that occur in nature or are chemically synthesized. There are over 140 known non-natural amino acids and potentially thousands of combinations. Examples of "non-natural" amino acids include 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, α-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 non-naturally occurring groups, groups or chemical moieties on the amino acid.

[0149] Reference to a specific amino acid is intended to encompass the amino acid in both conjugated and unconjugated states. Thus, the term "amino acid" refers to an amino acid residue in which part of its structure participates in the formation of a bond, which generally results in the loss of an atom of the amino acid itself, e.g., the equivalent of a water molecule when forming a peptide bond in a protein. The amino acid can be conjugated to the lipid moiety via any suitable functional group within the amino acid. In some embodiments, the amino acid is conjugated at its carboxyl terminus. In some embodiments, the amino acid is conjugated to the carbonyl carbon atom of a carboxylic acid residue.

[0150] In most cases, the names of naturally occurring and non-naturally occurring aminoacyl residues used herein follow the naming conventions recommended by the IUPAC Commission on the Nomenclature of Organic Chemistry and the IUPAC-IUB Commission on Biochemical Nomenclature, as set forth in "Nomenclature of α-Amino Acids (Recommendations, 1974)" Biochemistry, 14(2), (1975). If the names and abbreviations of amino acids and aminoacyl residues used in this specification and the appended claims differ from these recommendations, this will be clearly stated to the reader.

[0151] In this specification, unless the naturally occurring amino acids are referred to by their full names (e.g., alanine, arginine, etc.), they are represented by their conventional three-letter or single-letter abbreviations (e.g., Ala or A for alanine, Arg or R for arginine, etc.). As used herein, the term "L-amino acid" refers to the "L" isomeric form of a peptide, whereas the term "D-amino acid" refers to the "D" isomeric form of a peptide (e.g., Dphe, (D)Phe, D-Phe or D F is the D-isomeric form of phenylalanine). The D-isomeric form of an amino acid residue may replace any L-amino acid residue as long as the desired function is retained.

[0152] For less common or non-naturally occurring amino acids, unless referred to by their full names (e.g., sarcosine, ornithine, etc.), their residues are generally referred to by three- or four-letter codes, including Sar or Sarc (sarcosine, e.g., N-methylglycine), Aib (α-aminoisobutyric acid), Dab (2,4-diaminobutyric acid), Dapa (2,3-diaminopropionic 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).

[0153] In some embodiments, W is tryptophan, proline, lysine, histidine, arginine or a variant thereof. In some embodiments, W is tryptophan or a variant thereof.

[0154] In some embodiments, the compound of formula (II) is a compound of formula (IIa), wherein W is 1-methyl-D-tryptophan. In some embodiments, the compound of formula (IIa) is:

[0155]

[0156] or a pharmaceutically acceptable salt thereof.

[0157] 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 , where R 2 is selected from hydrogen and C1-C6 alkyl. In some embodiments, X is NH. In some embodiments, X is NR 2 , where R 2 In some embodiments, X is S.

[0158] In some embodiments, L is a lipid moiety having at least 8 carbon atoms. The lipid moiety can be derived from any suitable lipid, such as a fatty alcohol, a fatty acid, a phospholipid, a steroid or a synthetic lipid. In some embodiments, the lipid moiety is derived from a lipid having a functional group (such as a hydroxyl, a carboxylic acid group or an amino group), and the lipid moiety is connected 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 (II) is derived from a functional group; for example, if the lipid moiety is derived from a fatty alcohol, X is O.

[0159] In some embodiments, L is selected from 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.

[0160] For example, in some embodiments, L is selected from C8-C 80 Alkyl and C8-C 80 In some embodiments, L is selected from C8-C 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)-hexadeca-9-en-1-ol), oleyl alcohol ((Z)-octadeca-9-en-1-ol), anti-oleyl alcohol (trans-9-octadecaenol), cis-isooctadecaenol (cis-11-octadecaenol), gadol ((Z)-eicos-9-en-1-ol), 11-eicosene alcohol, erucyl alcohol (cis-13-eicosene alcohol), 15-eicosene-1-ol, eicosene alcohol (eicos-11,14-dien-1-ol), linolenic alcohol ((9Z, 12Z, 15Z)-9,1 2,15-octadecatriene-1-ol), γ-linolenic acid ((6E,9E,12E)-octadecatriene-6,9,12-triene-1-ol), tung oil alcohol (octadecatriene-9,11,13-triene-1-ol), eicosane-5,8,11-triene-1-ol, docosane-13-ene-1-ol, eicosane-11,14-17-triene-1-ol, octadecatriene-6,9,12,15-tetraene-1-ol, arachidonic acid ((5Z,8Z,11Z,14Z)-eicosatetraene-1-ol), 4E,6Z-hexadecadiene-1-ol, eicosane-5,8,11,14,17-pentaene-1-ol, docosahexaenol (docosahexaenol) ,7,10,13,16,19-hexaen-1-ol), 2-docosahexa-7,10,13,16,19-pentaen-1-ol, 2-tetradecanoic acid-6,9,12,15,18,21-hexaen-1-ol, n-octanol (1-octanol), nonanol (1-nonanol), decanol (1-decanol), undecanol (1-undecanol), lauryl alcohol (1-dodecanol), tridecanol (1-tridecanol), myristyl alcohol (1-tetradecanol), pentadecanol (1-pentadecanol), cetyl alcohol (1-hexadecanol), palmitoleyl alcohol (cis-9-hexadecene-1-ol), heptadecanol (1-n-heptadecanol), stearyl alcohol (1-octadecanol), oleyl alcohol (1-octadecenol), nonadecanol (1-nonadecanol), Arachidyl alcohol (1-eicosanol), heneicosanol (1-heneicosanol), behenyl alcohol (1-docosanol), erucyl alcohol (cis-13-docosan-1-ol), 1-tricosanol, tetracosanol (1-tetracosanol), pentacosanol (1-pentacosanol), hexacosanol (1-hexacosanol), 1-heptacosanol, montanol (1-octacosanol), 1-nonacosanol, myricanol (1-triacontanol), 1-triacontanol, 1-dotriacontanol (margarine alcohol), 1-triacontanol, candacetyl alcohol (1-tetratriacontanol), 1-hexatriacontanol, 1-heptatriacontanol, 1-octaticontanol, nonatriacontanol, 1-triacontanol, or 1-tetracontanol.

[0161] In some embodiments, L is selected from C8-C 80 Heteroalkyl, C8-C 80 Heteroalkenyl and C8-C 80 In such embodiments, L can be derived from a lipid comprising one or more heteroatom groups (such as -O-, -NH-, -C(O)-, etc.) or combinations thereof (e.g., a -C(O)O- group).

[0162] For example, in some embodiments, L has formula (A):

[0163]

[0164] in:

[0165] 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; and

[0166] R a and R b Each independently selected from C6-C 40 Alkyl, C6-C 40 Alkenyl, C6-C 40 Heteroalkyl and C6-C 40 Heteroalkenyl.

[0167] 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.

[0168] In some embodiments, R a and R b Each independently selected from C6-C 40 Alkyl and C6-C 40 For example, in some embodiments, R a and R bEach of the following is independently selected from the group consisting of n-hexyl, n-heptyl, n-octyl, n-nonyl, n-decyl, undecyl, dodecyl, tridecyl, tetradecyl, pentadecyl, hexadecyl, heptadecyl, octadecyl, nonadecyl, eicosyl, heneicosyl, docosyl, tricosyl, tetracosyl, pentacosyl, hexacosyl, heptacosyl, octacosyl, nonadecyl, triacontyl, triuncontyl, dotriacontyl, trictriacontyl, tetratriacontyl, pentatriacontyl, hexatriacontyl, heptatriacontyl, octatriacontyl, Nonadriatriadecyl, tetradecyl, linoleyl ((9Z,12Z)-octadec-9,12-dien-1-yl), palmityl ((Z)-hexadec-9-en-1-yl), oleyl ((Z)-octadec-9-en-1-yl), trans-oleyl (trans-9-octadecenyl), cis-11-octadecenyl (cis-vaccenyl) (cis-11-octadecenyl), gadyl ((Z)-eicos-9-en-1-yl), 11-eicosenyl, erucyl (cis-13-eicosenyl), 15 -Eicosyl, docosyl (eicosyl-11,14-dien-1-yl), linoleyl ((9Z,12Z,15Z)-9,12,15-octadecadien-1-yl), γ-linoleyl ((6E,9E,12E)-octadecadien-6,9,12-trien-1-yl), tung oil (octadecadien-9,11,13-trien-1-yl), eicosyl-5,8,11-trien-1-yl, docosyl-13-en-1-yl, eicosyl-11,14-17-trien-1-yl, In some embodiments, R a and R b Each is linoleyl.

[0169] In some embodiments, L is of formula (B) or (C):

[0170]

[0171] in:

[0172] 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;

[0173] R a1 Selected from C6-C 40 Alkyl and C6-C 40 alkenyl; and

[0174] R b Selected from C6-C 40 Alkyl, C6-C 40 Alkenyl, C6-C 40 Heteroalkyl and C6-C 40 Heteroalkenyl.

[0175] 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.

[0176] 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.

[0177] In some embodiments, R a1 C6-C 24 Alkyl or C6-C 24 In some embodiments, R a1 Selected from C9-C 22 Alkyl and C9-C 22 In some embodiments, R a1 Selected from straight or branched chain C 6、 C 7、 C 8、 C 9、 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、 C23、 or C 24 In some embodiments, R a1 Selected from straight or branched chain C 6、 C 7、 C 8、 C 9、 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 Selected from linoleyl, n-nonyl, n-undecyl, heneicosane-11-yl, pentadecane-7-yl and heptadecan-9-yl.

[0178] In some embodiments, R b Selected from C6-C 40 Alkyl and C6-C 40 For example, in some embodiments, R bis selected from the group consisting of n-hexyl, n-heptyl, n-octyl, n-nonyl, n-decyl, undecyl, dodecyl, tridecyl, tetradecyl, pentadecyl, hexadecyl, heptadecyl, octadecyl, nonadecyl, eicosyl, heneicosyl, docosyl, tricosyl, tetracosyl, pentacosyl, hexacosyl, heptacosyl, octacosyl, nonacosyl, triacontyl, triuncontyl, dotriacontyl, trictriacontyl, tetratriacontyl, pentatriacontyl, hexatriacontyl, heptatriacontyl, octatriacontyl, Nonatriacontyl, tetracontyl, linoleyl ((9Z, 12Z)-octadec-9,12-dien-1-yl), palmityl ((Z)-hexadec-9-en-1-yl), oleyl ((Z)-octadec-9-en-1-yl), transoleyl (trans-9-octadecenyl), cis-11-octadecenyl (cis-11-octadecenyl), gaddenyl ((Z)-eicos-9-en-1-yl), 11-eicosenyl, erucyl (cis-13-eicosenyl), 15-eicosene-1-yl, Docosyl (eicosyl-11,14-dien-1-yl), linoleyl ((9Z,12Z,15Z)-9,12,15-octadecatrien-1-yl), γ-linoleyl ((6E,9E,12E)-octadeca-6,9,12-trien-1-yl), tung oilyl (octadeca-9,11,13-trien-1-yl), eicosyl-5,8,11-trien-1-yl, eicosyl-13-en-1-yl, eicosyl-11,14-17-trien-1-yl, octadeca-6, In some embodiments, R b Linoleyl.

[0179] In some embodiments, L is of formula (D), (E), or (F):

[0180]

[0181] in:

[0182] 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; and

[0183] R a1 and R a2 Each independently selected from C6-C 40 Alkyl and C6-C 40 Alkenyl.

[0184] In some embodiments, L is of formula (D). In some embodiments, L is of formula (E). In some embodiments, L is of 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.

[0185] 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.

[0186] In some embodiments, R a1 and R a2 Each independently selected from C6-C 24 Alkyl and C6-C 24 In some embodiments, R a1 and R a2 Each independently selected from C9-C 22 Alkyl and C9-C 22 In some embodiments, R a1 and R a2 Each independently selected from a linear or branched C 6、 C 7、 C 8、 C 9、 C 10、 C 11、 C 12、 C 13、 C 14、 C 15、 C 16、 C 17、 C 18、 C 19、 C20、 C 21、 C 22、 C 23、 or C 24 In some embodiments, R a1 and R a2 Each independently selected from a linear or branched C 6、 C 7、 C 8、 C 9、 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 Each is independently selected from linoleyl, n-nonyl, n-undecyl, heneicos-11-yl, pentadec-7-yl and heptadecan-9-yl.

[0187] In some embodiments, L is derived from a steroid. For example, in some embodiments, L is derived from cholesterol, β-sitosterol, or BHEM-cholesterol.

[0188] In some embodiments, L is selected from:

[0189]

[0190]

[0191]

[0192]

[0193] The composition can be in the form of liposomes, lipid nanoparticles, micelles, etc. The method for preparing the lipid composition includes, for example, lipid film hydration, optionally combined with ultrasonic treatment or extrusion, solvent evaporation (for example, ethanol injection, ether injection or reverse phase evaporation), solvent diffusion method, thermal homogenization process, detergent removal method or a combination thereof. Any naturally occurring or synthetic vesicle-forming lipid or a combination thereof can be used. One or more vesicle-forming lipids can be selected from di-aliphatic chain lipids, such as phospholipids; diglycerides; di-aliphatic glycolipids; single lipids, such as sphingomyelin or sphingolipids; steroidal lipids; lipids derived from hydrophilic polymers; or mixtures thereof.

[0194] The lipid compositions of the present disclosure may include one or more cationic and / or ionizable lipids, phospholipids, neutral or non-cationic lipids, polyethylene glycol (PEG)-lipid conjugates, and / or structured lipids.

[0195] Cationic and / or ionizable lipids include, for example, amine-containing lipids that can be readily protonated and can have a positive or partially positive charge at physiological pH due to pKa values ​​between pH 5 and 8. The polar head groups of the cationic lipids preferably comprise amine derivatives such as primary, secondary and / or tertiary amines, quaternary ammonium, various combinations of amines, amidine salts or guanidine and / or imidazole groups as well as pyridinium, piperazine and amino acid head groups such as 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(tetradecyl)propyl-(2-hydroxyethyl)-dimethylammonium bromide (DMRIE), didodecyl(dimethyl)ammonium bromide (DDAB), 1,2-dioleyloxypropyl-3-dimethyl-hydroxyethylammonium 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).

[0196] In some embodiments, the disclosed compounds are incorporated into a lipid composition comprising one or more phospholipids. The phospholipids include a phospholipid moiety and one or more fatty acid moieties. The phospholipid moiety may include, but is not limited to, phosphatidylcholine, phosphatidylethanolamine, phosphatidylglycerol, phosphatidylserine, phosphatidic acid, 2-lysophosphatidylcholine and sphingomyelin. The fatty acid moiety may include, but is not limited to, lauric acid, myristic acid, myristoleic acid, palmitic acid, palmitoleic acid, stearic acid, oleic acid, linoleic acid, α-linolenic acid, erucic acid, phytanic acid, arachidic acid, arachidonic acid, eicosapentaenoic acid, behenic acid, docosapentaenoic acid and docosahexaenoic acid.

[0197] Phospholipids suitable for use in the composition may include, but are not limited to, phosphatidylglycerol (PG), including dimyristoylphosphatidylglycerol (DMPG) and 1,2-dioleoyl-sn-glycero-3-phospho-rac-(1-glycerol) sodium salt (DOPG); phosphatidylcholine (PC), including egg yolk phosphatidylcholine, dimyristoylphosphatidylcholine (DMPC), 1,2-distearoyl-sn-glycero-3-phosphocholine (DSPC), 1,2-dilinoleoyl-sn-glycero-3-phosphocholine (DLPC), 1,2-dioleoyl-sn-glycero-3-phosphocholine (DSPC), 1,2-dilinoleoyl-sn-glycero-3-phosphocholine (DLPC), 1,2-dioleoyl-sn-glycero-3-phosphocholine (DPSC ... 1,2-dipalmitoyl-sn-glycero-3-phosphocholine (DOPC), 1,2-diundecyl-sn-glycero-3-phosphocholine (DPPC), 1,2-diundecyl-sn-glycero-3-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-cholesteryl hemisuccinyl-sn-glycero-3-phosphocholine (OChemsPC), 1-hexadecyl-sn-glycero-3-phosphocholine (C16 Lyso PC), 1,2-dilinolenoyl-sn-glycero-3-phosphocholine, 1,2-di(eicosahexaenoyl)-sn-glycero-3-phosphocholine, 1,2-diarachidonoyl-sn-glycero-3-phosphocholine; phosphatidylethanolamine (PE) including 1,2-dioleoyl-sn-glycero-3-phosphoethanolamine (DOPE), 1,2-diphytanoyl-sn-glycero-3-phosphoethanolamine (ME 16.0PE), 1,2-distearoyl-sn-glycero-3-phosphoethanolamine, 1,2-dilinoleoyl-sn-glycero-3-phosphoethanolamine, 1,2-dialinolenoyl-sn-glycero-3-phosphoethanolamine, 1,2-diarachidonoyl-sn-glycero-3-phosphoethanolamine, 1,2-di(eicosahexaenoyl)-sn-glycero-3-phosphoethanolamine; phosphatidic acid (PA); phosphatidylinositol (PI); phosphatidylserine (PS); and sphingomyelin (SM).

[0198] In some embodiments, the lipid composition comprises a structured lipid. In some embodiments, the structured lipid is a sterol. Sterol can include cholesterol, coprosterol, ergosterol, campesterol, sitosterol, stigmasterol, brassicasterol or a sterol ester, such as cholesterol hemisuccinate, cholesterol sulfate, ursolic acid, alpha-tocopherol or any other cholesterol derivative.

[0199] In some embodiments, lipid compositions include polyethylene glycol (PEG)-lipid conjugates.PEG-lipid conjugates may include but are not limited to PEG-modified phosphatidylethanolamine, PEG-modified phosphatidic acid, PEG-modified ceramides, PEG-modified dialkylamines, PEG-modified diacylglycerols, PEG-modified dialkylglycerols, and mixtures thereof. For example, the PEG lipid can be PEG-DMG (1,2-dimyristoyl-rac-glycero-3-methoxypolyethylene glycol), PEG-c-DOMG (, 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-ditetradecylacetamide 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-ditetradecylacetamide.

[0200] The lipid structures described herein may also include other components that are typically used to form vesicles (e.g., for stabilization). Examples of such other components include, but are not limited to, fatty alcohols, fatty acids, and / or any other pharmaceutically acceptable excipients that can affect surface charge, membrane fluidity, and help incorporate lipids into lipid assemblies.

[0201] Lipid composition can also be targeted, for example, containing one or more targeting moieties or biodistribution modifiers on its surface. The targeting moiety can be any agent that can specifically bind to the desired target or interact, and is generally known in the art, for example, ligands such as folic acid, proteins, antibodies or antibody fragments, etc.). In some embodiments, the targeting moiety is an immune cell epitope (for example, B cell and T cell epitopes). In the embodiment selected, the targeting moiety comprises one or more epitopes from a microbial agent (for example, Clostridium difficile, Bacillus anthracis, Clostridium botulinum, Helicobacter pylori, rotavirus, Coronavirus family).

[0202] The lipid composition can have any structure, for example, a structure with an inner space isolated from an external medium by one or more lipid bilayers, or any microcapsule with a semipermeable membrane, wherein the semipermeable membrane has a lipophilic core, wherein the membrane isolates the interior. In some embodiments, the lipid composition can include a single-layer liposome, with a single lipid layer. In some embodiments, the lipid composition can include a liposome with a lipid bilayer. In some embodiments, the lipid composition includes micelles.

[0203] 4. Active agent

[0204] Any composition disclosed herein may also include at least one active agent. In some embodiments, at least one active agent includes nucleic acid (mRNA, aptamer, antisense oligonucleotide, ribozyme nucleic acid, interfering RNA, antisense nucleic acid and antigen nucleic acid), protein, immunomodulator, chemotherapeutic agent, steroid, analgesic, antibacterial agent or its combination. In some embodiments, at least one active agent is wrapped by liposome, lipid nanoparticle, micelle in lipid composition.

[0205] In some embodiments, at least one active agent comprises a nucleic acid (e.g., ribonucleic acid or deoxyribonucleic acid). Exemplary nucleic acids for use according to the present disclosure include, but are not limited to, one or more deoxyribonucleic acids (DNA), ribonucleic acids (RNA) (including 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, etc. In some embodiments, the nucleic acid encodes an antigen or a functional fragment thereof.

[0206] In some embodiments, at least one active agent is RNA. RNA that can be used 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), micro RNA (miRNA), Dicer substrate RNA (dsRNA), small hairpin RNA (shRNA), transfer RNA (tRNA), messenger RNA (mRNA) and mixtures thereof.

[0207] In certain embodiments, at least one active agent is mRNA. The mRNA can encode any polypeptide of interest, including any naturally or non-naturally occurring or otherwise modified polypeptide. The polypeptide encoded by the mRNA can be of any size and can have any secondary structure or activity. In some embodiments, the polypeptide encoded by the mRNA can have a therapeutic effect when expressed in a cell.

[0208] In other embodiments, at least one active agent is siRNA. siRNA may be able to selectively knock down or down-regulate the expression of a gene of interest. For example, after administering a nanoparticle composition comprising siRNA to a subject in need, siRNA may be selected to silence genes associated with a particular disease, disorder or condition. siRNA may comprise a sequence complementary to an mRNA sequence encoding a gene or protein of interest. In some embodiments, siRNA may be an immunomodulatory siRNA.

[0209] In some embodiments, at least one active agent is shRNA or a vector or plasmid encoding shRNA. After the appropriate construct is delivered to the nucleus, the shRNA can be produced in the target cell. The constructs and mechanisms associated with shRNA are well known in the relevant art.

[0210] 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, such as SM-36 and its analogs; toll-like receptor (TLR) agonists and analogs thereof, such as imiquimod, resiquimod, segamod, gademod, SM-360320, TMX-101, TMX-202, TMX-302, TMX-306, GSK2245035, CL097, 852A, AZD-8848, DSP-3025, GS-9620, RO7020, 531, 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, such as atorvastatin, pravastatin, fluvastatin, simvastatin, lovastatin, mevastatin, pitavastatin, rosuvastatin and their analogs.

[0211] In some embodiments, at least one active agent includes at least one chemotherapeutic agent. The term "chemotherapeutic agent" or "anticancer agent" as used herein includes any small molecule or other drug used for cancer treatment or prevention. Chemotherapeutic agents include, but are not limited to, cyclophosphamide, methotrexate, 5-fluorouracil, doxorubicin, docetaxel, daunorubicin, bleomycin, vinblastine, dacarbazine, cisplatin, paclitaxel, raloxifene hydrochloride, tamoxifen citrate, abemaciclib, everolimus, apellis, anastrozole, pamidronate, anastrozole, exemestane, capecitabine, epirubicin hydrochloride, eribulin mesylate, toremifene, fulvestrant, and sedrin. nt), letrozole, gemcitabine, goserelin, ixabepilone, emtansine, lapatinib, olaparib, megestrol acetate, neratinib, palbociclib, ribociclib, talazoparib, thiotepa, toremifene, methotrexate, and tucatinib.

[0212] In some embodiments, at least one active agent comprises an antimicrobial agent (e.g., an antiviral agent or a bactericidal agent). In some embodiments, the antimicrobial agent is an antiviral agent, including but not limited to abacavir, acyclovir, adefovir, amantadine, amprenavir, atazanavir, baloxavir marboxil), bictegravir, boceprevir, bulevirtide, cidofovir, cobicistat, daclatasvir, darunavir, delavirdine, didanosine, docosanol, dolutegravir, doravirine, edoxuridine, efavirenz, elvitegravir, emtricitabine, enfuvirtide, entecavir, etravirine, famciclovir, fomivirsen, fosamprenavir, foscarnet, ganciclovir, ibacitabine, ibalizumab, iodoxuridine, imiquimod, imunovir, indinavir, lamivudine, letermovir, lopinavir, loviramide, maraviroc araviroc), methisazone, morpholinoguanidine, nelfinavir, nevirapine, nexavir, nitazoxanide, oseltamivir, penciclovir, peramivir, penciclovir, peramivir, pleconaril, podophyllotoxin, raltegravir, remdesivir, ribavirin, rilpivirine, rilpivirine, rimantadine, rintatolimod, ritonavir, saquinavir, simeprevir, sofosbuvir, stavudine, taliveline, telaprevir, telbivudine, tenofovir (e.g., tenofovir alafenamide, alafenamide or tenofovir disoproxil), tipranavir, trifluridine, tromantine, umifenovir, valacyclovir, valganciclovir, vicriviroc, vidarabine, zalcitabine, zanamivir and zidovudine, and any combination thereof.

[0213] In some embodiments, the antibacterial agent is a bactericide. Exemplary bactericides include sulfonamides, amide alcohols, 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), polymyxins (e.g., polymyxin, colistin).

[0214] 5. Pharmaceutical Composition

[0215] The lipid composition can be incorporated into a pharmaceutically acceptable composition. Pharmaceutical compositions and formulations can include a pharmaceutically acceptable carrier. The term "pharmaceutically acceptable carrier" as used herein refers to a non-toxic, inert solid, semisolid or liquid filler, diluent, encapsulating material, surfactant, cyclodextrin or any type of formulation aid. Some examples of materials that can serve as pharmaceutically acceptable carriers are sugars, such as, but not limited to, lactose, glucose and sucrose; starches, such as, but not limited to, corn starch and potato starch; cellulose and its derivatives, such as, but not limited to, sodium carboxymethylcellulose, ethylcellulose and cellulose acetate; tragacanth; malt; gelatin; talc; excipients, such as, but not limited to, cocoa butter and suppository waxes; oils, such as, but not limited to, peanut oil, cottonseed oil, safflower oil, sesame oil, olive oil, corn oil and soybean oil; surfactants, such as, but not limited to, cremophor EL, cremophor RH 60, Solutol HS15 and polysorbate 80; cyclodextrins, such as but not limited to α-CD, β-CD, γ-CD, HP-β-CD, SBE-β-CD; glycols; such as propylene glycol; esters, such as but not limited to ethyl oleate and ethyl laurate; agar; buffers, such as but not limited to magnesium hydroxide and aluminum hydroxide; alginic acid; pyrogen-free water; isotonic saline; Ringer's solution; ethanol and phosphate buffer solutions, and other non-toxic compatible lubricants, such as but not limited to sodium lauryl sulfate and magnesium stearate, as well as colorants, release agents, coating agents, sweeteners, flavoring agents and fragrance enhancers, preservatives and antioxidants may also be present in the composition, according to the judgment of the formulator.

[0216] The route of administration of the composition and the form of the composition will determine the type of carrier to be used. The composition can be in a variety of forms, such as those suitable for systemic administration (e.g., oral, rectal, nasal, sublingual, buccal, implant, or parenteral injection) or local administration (e.g., dermal, pulmonary, nasal, otic, ocular, liposome delivery system, or iontophoresis).

[0217] Carriers for systemic administration generally include at least one of diluents, lubricants, binders, disintegrants, colorants, flavors, sweeteners, antioxidants, preservatives, glidants, solvents, suspending agents, wetting agents, surfactants, cyclodextrins, combinations thereof, etc. All carriers are optional in the composition.

[0218] Suitable diluents include sugars such as glucose, lactose, dextrose and sucrose; glycols such as propylene glycol; calcium carbonate; sodium carbonate; sugar alcohols such as glycerol; mannitol; and sorbitol. The amount of one or more diluents in a systemic or topical composition is generally about 50% to about 90%.

[0219] Suitable lubricants include silicon dioxide, talc, stearic acid and its magnesium and calcium salts, calcium sulfate; and liquid lubricants such as polyethylene glycol and vegetable oils such as peanut oil, cottonseed oil, sesame oil, olive oil, corn oil and cocoa butter. The amount of one or more lubricants in the systemic or local composition is generally about 5% to about 10%.

[0220] Suitable binders include polyvinyl pyrrolidone; magnesium aluminum silicate; starches such as corn starch and potato starch; gelatin; tragacanth; and cellulose and its derivatives such as sodium carboxymethylcellulose, ethylcellulose, methylcellulose, microcrystalline cellulose and sodium carboxymethylcellulose. The amount of one or more binders in the systemic composition is generally from about 5% to about 50%.

[0221] Suitable disintegrants include agar, alginic acid and its sodium salt, effervescent mixtures, cross-linked carboxymethylcellulose, cross-linked povidone, sodium carboxymethyl starch, sodium starch glycolate, clays and ion exchange resins. The amount of one or more disintegrants in the systemic or topical composition is generally from about 0.1% to about 10%.

[0222] Suitable coloring agents include colorants such as FD&C dyes.When used, colorants are generally present in systemic or topical compositions in amounts of from about 0.005% to about 0.1%.

[0223] Suitable flavors include menthol, mint, and fruit flavors.When used, the amount of one or more flavors in the systemic or topical compositions is generally from about 0.1% to about 1.0%.

[0224] Suitable sweeteners include aspartame and saccharin.The amount of one or more sweeteners in the systemic or topical compositions is generally from about 0.001% to about 1%.

[0225] Suitable antioxidants include butylated hydroxyanisole ("BHA"), butylated hydroxytoluene ("BHT"), and vitamin E. The amount of one or more antioxidants in the systemic or topical compositions is generally from about 0.1% to about 5%.

[0226] Suitable preservatives include benzalkonium chloride, methylparaben and sodium benzoate.The amount of one or more preservatives in the systemic or topical compositions is generally from about 0.01% to about 5%.

[0227] Suitable glidants include silicon dioxide.The amount of one or more glidants in the systemic or topical compositions is generally from about 1% to about 5%.

[0228] Suitable solvents include water, isotonic saline, ethyl oleate, glycerol, hydroxylated castor oil, alcohols such as ethanol, dimethyl sulfoxide, N-methyl-2-pyrrolidone, dimethylacetamide and phosphate (or other suitable buffers). The amount of one or more solvents in a systemic or topical composition is generally from about 0% to about 100%.

[0229] Suitable suspending agents include AVICEL RC-591 (from FMC Corporation of Philadelphia, Pa.) and sodium alginate. The amount of one or more suspending agents in the systemic or topical compositions is generally from about 1% to about 8%.

[0230] Suitable surfactants include lecithin, polysorbate 80 and sodium lauryl sulfate, and TWEENS from Atlas Powder Company of Wilmington, Del. Suitable surfactants include those disclosed in CTFA Cosmetic Ingredient Handbook, 1992, pp. 587-592; Remington's Pharmaceutical Sciences, 15th edition 1975, pp. 335-337; and McCutcheon, Vol. 1, Emulsifiers & Detergents, 1994, North American edition, pp. 236-239. The amount of one or more surfactants in the systemic or topical composition is generally from about 0.1% to about 5%.

[0231] Suitable cyclodextrins include α-CD, β-CD, γ-CD, hydroxypropyl β-cyclodextrin (HP-β-CD), sulfobutyl ether β-cyclodextrin (SBE-β-CD).The amount of cyclodextrin in the systemic or topical composition is generally from about 0% to about 40%.

[0232] Compositions for oral administration can have liquid form.For example, suitable liquid forms include aqueous solutions, emulsions, suspensions, solutions redissolved by non-effervescent granules, suspensions redissolved by non-effervescent granules, effervescent preparations redissolved by effervescent granules, elixirs, tinctures, syrups, etc.Liquid oral administration compositions generally include disclosed compounds and carriers, i.e., carriers selected from diluents, colorants, spices, sweeteners, preservatives, solvents, suspending agents and surfactants.Oral liquid compositions preferably include one or more components selected from colorants, spices and sweeteners.

[0233] Other compositions that can be used to achieve systemic delivery of the subject compound include sublingual, buccal and nasal dosage forms. Such compositions typically include one or more soluble filler materials, such as diluents, including sucrose, sorbitol and mannitol; and binders, such as gum arabic, microcrystalline cellulose, carboxymethyl cellulose and hydroxypropyl methylcellulose. Such compositions may also include lubricants, colorants, flavors, sweeteners, antioxidants and glidants.

[0234] The amount of carrier used in conjunction with the disclosed compositions is sufficient to provide a composition for the practical amount of compound administered per unit dose. Techniques and compositions for making 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).

[0235] A carrier may comprise a single ingredient or a combination of two or more ingredients.

[0236] 6. Vaccines

[0237] The compositions can also be used in vaccines. Vaccines comprise the disclosed compositions and antigens or nucleic acids encoding them. 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 can be used.

[0238] Depending on the infectious disease, autoimmune disease, disorder, cancer, pathogen, or condition being treated using a given vaccine composition, the antigens may be derived from and / or isolated from essentially any desired source.

[0239] The vaccines described herein may be capable of providing immunity against one or more conditions associated with an infectious disease, including but not limited to influenza, measles, human papillomavirus (HPV), rabies, meningitis, pertussis, tetanus, rubella, hepatitis, and tuberculosis, and may include infectious disease-derived antigens and / or epitopes, or nucleic acids encoding the same.

[0240] The vaccines described herein may also direct an immune response against cancer cells and may include tumor cell-derived antigens, epitopes and / or new epitopes, or portions thereof, or nucleic acids encoding tumor cell-derived antigens, epitopes and / or new epitopes. Tumor antigens are surface molecules differentially expressed by tumor cells relative to non-tumor tissues. Tumor antigens distinguish tumor cells from normal cells immunologically and provide diagnostic and therapeutic targets for human cancers. Tumor antigens have been characterized as altered carbohydrate molecules of membrane proteins or glycoproteins or glycolipids on the cell surface. Cancer cells often have unique tumor antigens on their surfaces, such as truncated epidermal growth factor, folate binding protein, epithelial mucin, melanoferrin, carcinoembryonic antigen, prostate-specific membrane antigen, HER2-neu, which are all candidates for therapeutic cancer vaccines. Because tumor antigens are normal or associated with normal components of the body, the immune system is generally unable to produce an effective immune response to those antigens to destroy tumor cells. Illustrative cancer types for which this method 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.

[0241] In other embodiments, the antigen present in the vaccine composition is not an exogenous 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, nervous system diseases, rheumatic diseases / connective tissue diseases, autoimmune cytopenias, and related autoimmune diseases. Such conventional organ-specific autoimmunity may include thyroiditis (Graves+Hashimoto's), gastritis, adrenalitis (Addison's), oophoritis, primary biliary cirrhosis, myasthenia gravis, gonadal failure, hypoparathyroidism, alopecia, malabsorption syndrome, pernicious anemia, hepatitis, anti-receptor antibody disease, and vitiligo. Such nervous system diseases may include schizophrenia, Alzheimer's disease, depression, hypopituitarism, diabetes insipidus, Sjögren's syndrome, and multiple sclerosis. Such rheumatic diseases / 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 retinitis, glomerulonephritis, post-myocardial infarction cardiotomy syndrome, pulmonary hemosiderosis, amyloidosis, sarcoidosis, aphthous stomatitis and other immune related diseases, as described herein and known in the relevant art.

[0242] In one embodiment, the antigen in the vaccine composition is a peptide, polypeptide or an immunogenic portion thereof. As used herein, an "immunogenic portion" is a protein portion that is recognized (e.g., specifically bound) by a B cell and / or T cell surface antigen receptor. Such an immunogenic portion typically comprises at least 5 amino acid residues of the antigen protein or a variant thereof, more preferably at least 10, and even more preferably at least 20 amino acid residues.

[0243] The immunogenic portion of an antigenic polypeptide can be generally identified using well-known techniques, such as those summarized in Paul, Fundamental Immunology, 3rd edition, 243-247 (Raven Press, 1993) and the references cited therein. Such techniques include screening polypeptides for the ability to react with antigen-specific antibodies, antisera and / or T cell lines or clones. As used herein, if antisera and antibodies specifically bind to an antigen (e.g., they react with proteins in ELISA or other immunoassays, and do not react detectably with unrelated proteins), they are "antigen-specific". Such antisera and antibodies can be prepared using known techniques. The immunogenic portion of a protein is a portion that reacts with such antisera and / or T cells, and its reaction level is not significantly lower than the reactivity of the full-length polypeptide (e.g., in ELISA and / or T cell reactivity assays). Such immunogenic portions can react at a level similar to or higher than the reactivity of the full-length polypeptide in such assays. Such screening can generally be performed using methods well known to those of ordinary 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 can be immobilized on a solid support and contacted with patient serum to allow antibodies in the serum to bind to the immobilized polypeptide. Unbound serum can then be removed and the antibody can be screened using, for example, 125 Bound antibodies were detected by I-labeled protein A.

[0244] Peptide and polypeptide antigens can be prepared using any of a variety of well-known techniques. Recombinant polypeptides encoded by DNA sequences can be easily prepared from isolated DNA sequences using any of a variety of expression vectors known to those of ordinary skill in the art. Expression can be achieved in any appropriate host cell that has been transformed or transfected with an expression vector containing a DNA molecule encoding the recombinant polypeptide. Suitable host cells include prokaryotes, yeast, and higher eukaryotic cells, such as mammalian cells and plant cells. Preferably, the host cell used is an E. coli, yeast, or mammalian cell line, such as COS or CHO.

[0245] Portions and other variants of protein antigens having less than about 100 amino acids and generally less than about 50 amino acids may also be produced by synthetic methods using techniques well known to those of ordinary skill in the art. For example, such polypeptides may be synthesized using any commercially available solid phase technique (such as the Merrifield solid phase synthesis method, in which amino acids are sequentially added to a growing amino acid chain). See, Merrifield, J. Am. Chem. Soc. 85: 2149-2146, 1963. Equipment for automatic synthesis of polypeptides is commercially available from suppliers such as Perkin Elmer / Applied BioSystems Division (Foster City, Calif.), and may be operated according to the manufacturer's instructions.

[0246] In certain embodiments, the nucleic acid encoding the antigen is DNA. This type of illustrative DNA-based vaccine contains DNA encoding one or more polypeptide antigens so that the antigen is produced in situ. Alternatively, the vaccine can be an RNA-based vaccine. In certain embodiments, the nucleic acid encoding the antigen is mRNA. mRNA can encode any polypeptide antigen of interest, including any natural or non-naturally occurring or otherwise modified polypeptide. The polypeptide encoded by mRNA can be of any size and can have any secondary structure or activity. In some embodiments, the polypeptide encoded by mRNA can stimulate an immune response when expressed in a cell.

[0247] Vaccine compositions of the present disclosure may also contain other compounds, which may be biologically active or inactive. Vaccines or drugs may include adjuvants or immunostimulants, or polynucleotides encoding adjuvants or immunostimulants (e.g., adjuvant polypeptides). Adjuvants and immunostimulants are compounds or compositions that directly or indirectly stimulate the immune system's response to co-administered antigens. In some embodiments, vaccines are non-adjuvanted or self-adjuvanting.

[0248] Suitable adjuvants are commercially available, for example, glucose-based lipid adjuvant (GLA); Pam3CSK4; Freund's incomplete adjuvant 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 such as aluminum hydroxide gel (alum), AlK(SO4)2, AlNa(SO4)2, AlNH4(SO4), and Al(OH)3; calcium salts (e.g., Ca3(PO4)2), iron salts, or zinc salts; insoluble suspensions of acylated tyrosine; acylated sugars; cationic or anionic derivatized polysaccharides; polynucleotides (e.g., polyIC, polyAU, and CpG oligodeoxynucleotides (e.g., class A or class B)); polyphosphazenes; cyanoacrylates; polymerase-(DL-lactide-co-glycoside); bovine serum albumin; diphtheria toxoid; tetanus toxoid; erythrine; keyhole-limpet hemocyanin; Pseudomonas toxin A; cholera toxoid; cholera toxin; pertussis toxin; viral proteins; Quil A; aminoalkylglucosamine phosphate compounds. In addition, adjuvants such as cytokines (e.g., GM-CSF or interleukin-2, -7 or -12), interferons or tumor necrosis factor 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 skilled in the art. When obtained from recombinant sources, the adjuvant may comprise a protein fragment comprising at least the immunostimulatory portion of the molecule.

[0249] Other known immunostimulatory macromolecules that can be used include, but are not limited to, polysaccharides, tRNA, non-metabolizable synthetic polymers such as polyvinylamine, polymethacrylic acid, polyvinylpyrrolidone, mixed condensation polymers of 4',4-diaminodiphenylmethane-3,3'-dicarboxylic acid and 4-nitro-2-aminobenzoic acid (having relatively high molecular weight) (see Sela, M., Science 166: 1365-1374 (1969)), or glycolipids, lipids, or carbohydrates.

[0250] In some embodiments, adjuvant polypeptides include immune activating proteins, such as CD70, CD40 ligands and constitutively active TLR4, or polycationic peptides (e.g., protamine). In some embodiments, the adjuvant polypeptide is a flagellin polypeptide. Commercially available mRNA encoding adjuvant polypeptides can be used as, for example, 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 an anti-Müllerian hormone receptor II extracellular domain (AMHR2-ED), as described above, and the other encoding an adjuvant polypeptide (e.g., a flagellin polypeptide or an immune activating protein).

[0251] Vaccine preparation is a mature art, and general guidance for vaccine preparation and formulation is readily available from any of a variety of sources. One such example is New Trends and Developments in Vaccines, ed. Volier et al. University Park Press, Baltimore, Md., USA 1978. Vaccine compositions can generally be used for both prophylactic and therapeutic purposes.

[0252] The amount of antigen in each dose of vaccine is generally selected to induce an immunoprotective response without significant adverse side effects in typical vaccines. This amount will vary depending on the specific immunogen used and its presentation. Of course, the dose administered may be based on age, body weight, the type of concurrent treatment (if any) and the nature of the antigen administered.

[0253] The immunogenic activity of a given amount of a vaccine composition can be readily determined, for example, by monitoring increases in antibody titers against the antigen used in the vaccine composition (Dalsgaard, K. Acta Veterinia Scandinavica 69: 1-40 (1978)). Another common method involves injecting various amounts of the vaccine composition intradermally into CD-1 mice, after which serum is harvested from the mice and tested for anti-immunogen antibodies, for example, by ELISA. These and other similar methods will be apparent to those skilled in the art.

[0254] 7. How to use

[0255] The present disclosure provides a method for delivering an active agent to a cell. The method includes contacting the cell with a composition as disclosed herein. In some embodiments, the cell is in vitro. In some embodiments, the cell is in vivo. Therefore, the present disclosure also provides a method for delivering any active agent to a subject. The method may include administering a composition as described herein to a subject.

[0256] The present disclosure also provides a method for producing a polypeptide of interest in a cell. The method includes contacting the cell with a composition as disclosed herein, wherein the composition includes mRNA encoding a polypeptide of interest. In some embodiments, the cell is in vitro. In some embodiments, the cell is in vivo. Therefore, the present disclosure provides a method for producing a polypeptide of interest in a subject, comprising administering a composition as described herein to the subject, wherein the composition includes mRNA encoding a polypeptide of interest.

[0257] The present disclosure also provides methods for treating a disease or disorder comprising administering to a subject in need thereof a composition as disclosed herein. In some embodiments, the subject is a human.

[0258] Diseases or conditions may include cancer, autoimmune diseases, inflammatory diseases, and infectious diseases.

[0259] In some embodiments, the disease or condition is an inflammatory disease or condition. The feature of an inflammatory disease is that the immune system in a tissue or organ is activated to an abnormal level, which may lead to abnormal tissue or organ function and / or disease. Inflammatory diseases and conditions treatable 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, Sjögren's syndrome or Churg-Strauss syndrome.

[0260] In some embodiments, the disease or disorder is an autoimmune disease or disorder.Autoimmune diseases and disorders refer to conditions in a subject characterized by damage to cells, tissues, and / or organs caused by an immune response of the subject to its own cells, tissues, and / or organs. Autoimmune diseases and conditions treatable by the methods of the invention include, but are not limited to, alopecia areata, ankylosing spondylitis, antiphospholipid syndrome, autoimmune Addison's disease, adrenal autoimmune disease, autoimmune hemolytic anemia, autoimmune hepatitis, autoimmune oophoritis and orchitis, autoimmune thrombocytopenia, Behcet's disease, bullous pemphigoid, cardiomyopathy, celiac dermatitis, chronic fatigue immune dysfunction syndrome (CFIDS), chronic inflammatory demyelinating polyneuropathy, Churg-Strauss syndrome, cicatricial pemphigoid, CREST syndrome, cold agglutinin disease, Crohn's disease, discoid lupus, essential mixed cryoglobulinemia, fibromyalgia-fibromyositis, glomerulonephritis, Graves' disease, Guillain-Barre syndrome, Hashimoto's thyroiditis, idiopathic pulmonary fibrosis, idiopathic thrombocytopenia, and inflammatory dysregulation of the thyroid gland. Indolent purpura (ITP), irritable bowel syndrome (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 syndromes, polymyalgia rheumatica, polymyositis and dermatomyositis, primary agammaglobulinemia, primary biliary cirrhosis, psoriasis, psoriatic arthritis, Raynaud's phenomenon, Reiter's syndrome, rheumatoid arthritis, sarcoidosis, scleroderma, Sjögren's syndrome, myotonia solium, systemic lupus erythematosus, lupus erythematosus, Takayasu's arteritis, temporal arteritis / giant cell arteritis, ulcerative colitis, uveitis, vasculitis such as dermatitis herpetiformis vasculitis, vitiligo, and Wegener's granulomatosis.

[0261] 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 methods of the invention include, but are not limited to, asthma, encephalitis, inflammatory bowel disease, chronic obstructive pulmonary disease (COPD), allergic disorders, pulmonary fibrosis, undifferentiated spondyloarthropathies, undifferentiated arthropathy, arthritis, inflammatory osteolysis, and chronic inflammation caused by chronic viral or bacterial infections. Examples of psoriasis types that can be treated according to the compositions and methods of the invention include, but are not limited to, plaque psoriasis, pustular psoriasis, erythrodermic psoriasis, guttate psoriasis, and inverse psoriasis.

[0262] In some embodiments, the disease or condition is cancer. In some embodiments, cancer comprises solid tumors. In some embodiments, cancer comprises blood cancer or lymphoma. In some embodiments, cancer is metastatic cancer. In some embodiments, the disclosed compounds, compositions or methods result in the elimination of inhibition of metastasis. 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.

[0263] The compounds and compositions herein can be used to treat various cancers, including carcinomas, sarcomas, lymphomas, leukemias, melanomas, mesotheliomas, multiple myeloma, or seminoma. The cancer can be bladder cancer, blood cancer, bone cancer, brain cancer, breast cancer, cervical cancer, colon / rectum cancer, endometrial cancer, head and neck cancer, kidney cancer, liver cancer, lung cancer, lymph node cancer, muscle tissue cancer, ovarian cancer, pancreatic cancer, prostate cancer, skin cancer, spleen cancer, stomach cancer, testicular cancer, thyroid cancer, or uterine cancer.

[0264] 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.

[0265] In some embodiments, the disease or condition is an infectious disease. Infectious diseases that can be treated or prevented by the methods of the present invention are caused by infectious sources, including but not limited to viruses, bacteria, fungi, protozoa, worms 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, such as Mycobacterium tuberculosis, Chlamydia, Francisella tularensis; DNA viruses, such as Herpesviridae (Herpes simplex virus-1, Kaposi's sarcoma-associated virus and Epstein-Barr virus), Papillomaviridae (Human papillomavirus), Adenovirus and Hepadnaviridae (Hepatitis B virus) or RNA viruses, such as Retroviridae (Human immunodeficiency virus), Flaviviridae (Dengue virus, Hepatitis C virus), Orthomyxoviridae (Influenza) and Coronaviridae (Human coronavirus and SARS coronavirus).

[0266] The compositions disclosed herein can be administered to a subject by various methods. In any use or method described herein, the compositions can be administered by various routes known to those skilled in the art, including but not limited to oral, inhalation, intravenous, intramuscular, topical, subcutaneous, systemic and / or intraperitoneal administration to a subject in need thereof.

[0267] The amount of the composition of the present disclosure that needs to be used in the methods described herein varies not only with the specific compound selected, but also with the route of administration, the nature and / or symptoms of the disease, and the age and illness of the patient, and will ultimately be determined by the attending physician or clinician. The determination of the effective dosage level (i.e., the dosage level required to achieve the desired result) can be accomplished by those skilled in the art using conventional methods (e.g., human clinical trials, in vivo studies, and in vitro studies). For example, a useful dosage can be determined by comparing its in vitro activity with its in vivo activity in an animal model.

[0268] The amount and interval of the dosage can be adjusted individually to provide a plasma level of the active part sufficient to maintain the modulatory effect, or the minimum effective concentration (MEC). The MEC of each compound will vary, but can be estimated from in vivo and / or in vitro data. The dosage required to achieve the MEC will depend on individual characteristics and route of administration. However, FIPLC assays or bioassays can be used to determine plasma concentrations. Dosage intervals can also be determined using MEC values. The composition should be administered using a regimen that maintains plasma levels above the MEC for 10-90% of the time, preferably between 30-90%, and most preferably between 50-90%. In the case of local administration or selective uptake, the effective local concentration of the drug may be independent of the plasma concentration.

[0269] It should be noted that the attending physician will know how and when to terminate, interrupt or adjust administration due to toxicity or organ dysfunction. Conversely, if the clinical response is insufficient (excluding toxicity), the attending physician will also know to adjust the treatment to a higher level. In the management of the condition of interest, the size of the administered dose will vary with the severity of the symptoms to be treated and the route of administration. In addition, the dose, and perhaps the dose frequency, will also vary according to the age, weight and response of the individual patient. Programs comparable to those discussed above can be used in veterinary medicine.

[0270] Compositions disclosed herein can be evaluated for their efficacy and toxicity using known methods. For example, the toxicology of a particular compound or a subset of compounds sharing certain chemical moieties or their compositions can be determined by determining in vitro toxicity to a cell line (such as a mammalian cell line, and preferably a human cell line). The results of such studies can generally predict toxicity in animals (such as mammals or more specifically humans). Alternatively, the toxicity of a particular compound in an animal model, such as a mouse, rat, rabbit, dog or monkey model, can be determined using known methods. Several recognized methods can be used to determine the efficacy, such as in vitro methods, animal models or human clinical trials. When selecting a model to determine the efficacy, those skilled in the art can select an appropriate model, dosage, route of administration and / or regimen under the guidance of the prior art.

[0271] A variety of second therapies can be used in combination with the compounds and methods of the present disclosure. The second therapy can be the administration of an additional active agent, or can be a second therapy unrelated to the administration of another agent. Such second therapies include, but are not limited to, surgery, immunotherapy, radiotherapy.

[0272] The second therapy can be administered simultaneously with the initial therapy, or in the same composition, or in a separate composition that is administered substantially simultaneously with the first composition. In some embodiments, the second therapy can precede or follow treatment with the first therapy with a time interval ranging from a few hours to several months.

[0273] In some embodiments, the additional active agents include immunomodulators, chemotherapeutic agents, nucleic acids (e.g., mRNA, aptamers, antisense oligonucleotides, ribozyme nucleic acids, interfering RNA, antigene nucleic acids), steroids, analgesics, and antimicrobial agents, or combinations thereof.

[0274] Exemplary immunomodulators include: signal transducer and activator of transcription 3 (Stat3) inhibitors and analogs thereof, such as SM-36 and its analogs; toll-like receptor (TLR) agonists and analogs thereof, such as imiquimod, resiquimod, segamod, gademod, SM-360320, TMX-101, TMX-202, TMX-302, TMX-306, GSK2245035, CL097, 852A, AZD-8848, DSP-3025, GS-9620, RO7020 531, 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, such as atorvastatin, pravastatin, fluvastatin, simvastatin, lovastatin, mevastatin, pitavastatin, rosuvastatin and their analogs.

[0275] In some embodiments, the additional active agent includes at least one chemotherapeutic agent.As used herein, the term "chemotherapeutic drug" or "anti-cancer drug" includes any small molecule or other drug used for cancer treatment or prevention. Chemotherapeutic drugs include but are not limited to cyclophosphamide, methotrexate, 5-fluorouracil, doxorubicin, docetaxel, daunorubicin, bleomycin, vinblastine, dacarbazine, cisplatin, paclitaxel, raloxifene hydrochloride, tamoxifen citrate, abemaciclib, afinitor (everolimus), apellis, anastrozole, pamidronate, anastrozole, exemestane, capecitabine, epirubicin hydrochloride, eribulin mesylate, toremifene, fulvestrant, letrozole, gemcitabine, goserelin, ixabepilone, imatinib, lapatinib, olaparib, megestrol acetate, neratinib, palbociclib, ribociclib, talazopanib, thiotepa, toremifene, methotrexate and tucatinib. In selected embodiments, the chemotherapeutic agent includes paclitaxel.

[0276] In some embodiments, the composition can be co-administered with an antibacterial agent (e.g., an antiviral agent or a bactericide). In some embodiments, the antibacterial agent is an antiviral agent, including but not limited to abacavir, acyclovir, adefovir, amantadine, amprenavir, atazanavir, baloxavir dipivoxil, bitravir, boceprevir, brevitide, cidofovir, cobicistat, daclatasvir, darunavir, delavirdine, didanosine, docosanol, dolutegravir, doravirine, edouridine, efavirenz, elvitegravir, emtricitabine, enfuvirtide, entecavir, etravirine, famciclovir, fomivirsen, fosamprenavir, foscarnet, ganciclovir, ibacitabine, ibalizumab, iodoxuridine, imiquimod, imunovir, indinavir, lamivudine, letermovir, lopinavir, loviramide, Maraviroc, mefenamic acid, morphine, nelfinavir, nevirapine, nesavir, nitazoxanide, oseltamivir, penciclovir, peramivir, penciclovir, peramivir, pruconaril, podophyllotoxin, raltegravir, remdesivir, ribavirin, rilpivirine, rilpivirine, rimantadine, lintamod, ritonavir, saquinavir, simeprevir, sofosbuvir, stavudine, tarivudine, telaprevir, telbivudine, tenofovir (e.g., tenofovir alafenamide or tenofovir disoproxil), tipranavir, trifluridine, tromantanide, umifenovir, valacyclovir, valganciclovir, viraviroc, vidarabine, zalcitabine, zanamivir and zidovudine, and any combination thereof.

[0277] In some embodiments, the antibacterial agent is a bactericide. Exemplary bactericides include sulfonamides, amide alcohols, 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), polymyxins (e.g., polymyxin, colistin).

[0278] In some embodiments, the second therapy comprises immunotherapy. Immunotherapy includes chimeric antigen receptor (CAR) T cells or T cell transfer therapy, cytokine therapy, immunomodulators, cancer vaccines, or administration of antibodies (e.g., monoclonal antibodies).

[0279] In some embodiments, immunotherapy includes administering antibodies. Antibodies can target 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 known as erbB or EGFR), VEGF, CTLA-4 (also known as CD152), epithelial cell adhesion molecule (EpCAM, also known as 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, etc.). 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 linked to a chemotherapeutic agent. Thus, in some embodiments, the antibody is an antibody-drug conjugate.

[0280] Immunotherapy (e.g., administration of antibodies) can be administered to a subject by a variety of methods. In any of the uses or methods described herein, various routes known to those skilled in the art can be used, including but not limited to oral, inhaled, intravenous, intramuscular, topical, subcutaneous, systemic and / or intraperitoneal administration to a subject in need. Immunotherapy can be administered by parenteral administration (including but not limited to subcutaneous, intramuscular, intravenous, intraperitoneal, intracardiac and intraarticular injection). In some embodiments, immunotherapy can be administered in the same or different manner as the compounds or compositions disclosed herein.

[0281] 8. Test kit

[0282] On the other hand, the present disclosure provides a kit comprising at least one disclosed compound or a pharmaceutically acceptable salt thereof, or a composition comprising the compound or a pharmaceutically acceptable salt thereof, and instructions for using the compound or composition. In some embodiments, the kit includes at least one disclosed compound and one or more cations and / or ionizable lipids, phospholipids, neutral or non-cationic lipids, polyethylene glycol (PEG)-lipid conjugates and / or structural lipids. The kit may also include at least one active agent. The kit may also include other agents and / or products that are co-packaged, co-formulated and / or co-delivered with other components.

[0283] The kit may also include instructions for using the kit components. Instructions are related materials or methods related to the kit. Materials may include any combination of background information, component lists, brief or detailed schemes for using the composition, problem solving, references, technical support, and any other related documents. Instructions may be provided with the kit or as separate element components (as written or electronic form that can be provided on a computer-readable storage device or downloaded from an Internet website), or as a recorded demonstration.

[0284] It should be understood that the disclosed kits can be used in combination with the disclosed methods. The kits can also include containers or devices for use with the methods or compositions disclosed herein. The kits can optionally provide additional components, such as buffers and disposable equipment (e.g., pipettes, cell culture plates or flasks).

[0285] The kits provided herein are packaged in suitable packaging. Suitable packaging includes, but is not limited to, vials, bottles, jars, soft packaging, etc. The components of the kit can be physically packaged together or individually.

[0286] 9. Examples

[0287] The abbreviations used in the following schemes and examples are: BOC is tert-butyloxycarbonyl; DMAP is 4-dimethylaminopyridine; DMF is dimethylformamide; EDC is 1-ethyl-3-(3-dimethylaminopropyl)carbodiimide; eq is equivalent; EtOAc is ethyl acetate; MeOH is methanol; RT or rt is room temperature; and THF is tetrahydrofuran.

[0288] All air- and moisture-sensitive manipulations were performed under argon or 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 noted.

[0289] Analytical thin layer chromatography (TLC) was performed using Merck SIL G / UV254 plates. Compounds were visualized by exposure to UV light or immersion of the plates in ninhydrin or potassium permanganate solution followed by heating or staining with iodine vapor in a wide pot chamber. Column chromatography was performed in air using silica gel 60 (Fluka). Column chromatography was performed using Merck Kieselgel 60 (200-500 mm). Solvent systems (s / sv:v) are given.

[0290] NMR spectra 1H (300 MHz), 13C (75 MHz) were recorded on ARX 300 or Avance II 500 Bruker spectrometers, respectively. Chemical shifts (δ, ppm) are given with reference to the residual 1H or 13C of the deuterated solvent in the indicated solvent (CDCl3 7.26, 77.00; (CD3)2CO 2.05, 29.84 and 206.26, (CD3)2SO 2.50, 39.52). 1H- and 13C-NMR chemical shifts (δ) are in parts per million (ppm) relative to the TMS scale. Coupling constants J are in Hz. The following abbreviations are used for proton spectra multiplicities: s: singlet, d: doublet, t: triplet, q: quartet, qt: quintet, m: multiplet, br.: broad, dd: doublet of doublets, dt: doublet of triplets. Coupling constants (J) are reported in Hertz (Hz). Some unattributable signals will be indicated by ArH (aromatic hydrogen).

[0291] Mass spectra (MS) were recorded using a LCQ-advantage (ThermoFinnigan) mass spectrometer with positive (ESI+) or negative (ESI-) electrospray ionization (ionization tension 4.5 kV, injection temperature 240°C).

[0292] Example 1

[0293] Compound

[0294] General procedure 1: esterification. Under nitrogen atmosphere, a solution of carboxylic acid derivative (1eq), EDC hydrochloride (1eq) and DMAP (0.5eq) in anhydrous THF (0.1M) was stirred at 0°C for 0.5h. A solution of alcohol (1eq) in anhydrous THF (0.1M) was then added. The solution was stirred at 0°C to rt for 16h. The reaction progress was monitored by TLC using a CH2Cl2-MeOH 10:1 mixture as eluent and the 1H NMR using an ARX 300Brucker spectrometer. After the reaction was completed, the mixture was extracted with dichloromethane (3x). The organic layers were combined and washed with 0.2M HCl aqueous solution, saturated NaHCO3 aqueous solution and brine, respectively. Then, the solid residue was purified by manual column chromatography using a CH2Cl2-EtOAc or CH2Cl2-MeOH step gradient solvent system as eluent to obtain the title product.

[0295] (9Z,12Z)-Octadecane-9,12-dien-1-yl 1-methyl-D-tryptophanate

[0296]

[0297] Step 1: Follow General Procedure 1 at room temperature to obtain a α Na-(tert-butoxycarbonyl)-1-methyl-D-tryptophanate was synthesized from (9Z,12Z)-octadec-9,12-dien-1-yl Na-(tert-butoxycarbonyl)-1-methyl-D-tryptophan (1 eq), EDC.hydrochloride (1 eq.), DMAP (0.5 eq.) and linoleyl alcohol (1 eq.) in anhydrous THF (0.1 M) over a period of 4 h.

[0298] Step 2: (9Z,12Z)-octadec-9,12-dien-1-yl 1-methyl-D-tryptophanate was synthesized from a stirred solution of (9Z,12Z)-octadec-9,12-dien-1-yl Na-(tert-butoxycarbonyl)-1-methyl-D-tryptophanate (1 eq) in dioxane HCl (HCl 4 M in dioxane) over 5 h at 0 °C to RT.

[0299] ((4-((1-methyl-D-tryptophanyl)oxy)butyl)azanediyl)bis(hexane-6,1-diyl)bis(2-hexyldecanoate)

[0300]

[0301] Step 1: Follow General Procedure 1 at room temperature to obtain a α-(tert-Butyloxycarbonyl)-1-methyl-D-tryptophan (1 eq), EDC.hydrochloride (1 eq.), DMAP (0.5 eq.) and ALC-0315 (1 eq) in anhydrous THF (0.1 M) were synthesized over 4 h with ((4-((1-methyl-Na-pivaloyl-D-tryptophanyl)oxy)butyl)azanediyl)bis(hexane-6,1-diyl)bis(2-hexyldecanoate).

[0302] Step 2: ((4-((1-methyl-D-tryptophyl)oxy)butyl)azanediyl)bis(hexane-6,1-diyl)bis(2-hexyldecanoate) was synthesized from a stirred solution of ((4-((1-methyl-Na-pivaloyl-D-tryptophyl)oxy)butyl)azanediyl)bis(hexane-6,1-diyl)bis(2-hexyldecanoate) (1 eq) in dioxane HCl (HCl 4M in dioxane) over 5 h at 0 °C to RT.

[0303] 1 H NMR (500MHz, CDCl3) δ7.59(d,J=7.9Hz,1H),7.28(d,J=8.2Hz,1H),7.21(t,J=7.5H z,1H),7.09(t,J=7.4Hz,1H),6.92(s,1H),4.16–4.00(m,8H),3.74(s,3H),3.26(d d,J=14.4,4.8Hz,1H),3.01(dd,J=14.4,7.8Hz,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=6.8Hz,12H).

[0304] Heptadecan-9-yl 8-((4-((1-methyl-D-tryptophanyl)oxy)butyl)(6-oxo-6-(undecyloxy)hexyl)amino)octanoate

[0305]

[0306] Step 1: Follow General Procedure 1 at room temperature to obtain a α -(tert-Butyloxycarbonyl)-1-methyl-D-tryptophan (1 eq), EDC. hydrochloride (1 eq.), DMAP (0.5 eq.) and SM-102 (1 eq) in anhydrous THF (0.1 M) were synthesized over 4 h in 9-heptadecanyl 8-((4-((1-methyl-Na-pivaloyl-D-tryptophanyl)oxy)butyl)(6-oxo-6-(undecyloxy)hexyl)amino)octanoate.

[0307] Step 2: Heptadecan-9-yl 8-((4-((1-methyl-tryptophyl)oxy)butyl)(6-oxo-6-(undecyloxy)hexyl)amino)octanoate was synthesized from a stirred solution of heptadecan-9-yl 8-((4-((1-methyl-Na-pivaloyl-D-tryptophyl)oxy)butyl)(6-oxo-6-(undecyloxy)hexyl)amino)octanoate (1 eq) in dioxane HCl (HCl 4M in dioxane) at 0°C to RT during 5 h.

[0308] 1 H NMR (500MHz, CDCl3) δ7.58(d,J=7.9Hz,1H),7.27(d,J=7.7Hz,1H),7.20(t,J=7.5Hz,1H),7.09(t,J=7.4Hz,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=1 4.4, 4.8Hz, 1H), 3.02 (dd, J = 14.4, 7.6Hz, 1H), 2.62 (t, J = 6.4Hz, 2H), 2.40 (dt, J = 8.0, 5.3Hz, 4H), 2.26 (dt, J = 10. 6,7.5Hz,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=7.0Hz,9H).

[0309] Example 2

[0310] Lipid composition

[0311] Exemplary lipid compositions include any or all of the following: nucleic acids (such as mRNA, aptamers, antisense oligonucleotides, ribozyme nucleic acids, RNA interference, and antigene nucleic acids); modified lipids as disclosed herein; phospholipids (e.g., phosphatidylcholine and phosphatidylethanolamine); cholesterol; and polyethylene glycol (PEG)-functionalized lipids (PEG-lipids).

[0312] Cationic liposomes were generated using the dry film method. Lipids and cholesterol were dissolved in chloroform in a round-bottom flask. After removing the organic solvent by rotary evaporation at 40°C, the dried lipid solids were then hydrated in PBS (pH 7.4) and vortexed six times for 30 seconds every 5 min at 45°C. The resulting lipid suspension was subjected to 10 freeze-thaw cycles between liquid nitrogen and a 37°C water bath and extruded through an extruder with a 0.2 μm polycarbonate filter to obtain a uniform nanosuspension. The nucleic acid was then mixed with the liposomes and incubated for 20 min at RT to allow for adequate encapsulation.

[0313] Prepare nucleic acids in acetate buffer at pH 4.0. Prepare ethanol solutions containing the appropriate molar ratio of lipids depending on the desired formulation.

[0314] The diameter, size distribution, polydispersity index (PDI), and zeta potential were determined using a ZETASIZER Nano (ZEN3600, Malvern Instruments, Malvern, WR, UK), and the morphology of lipid nanoparticles and liposomes was visualized using a JEOL2010F transmission electron microscope (TEM) and a FEIQuanta 200FEG scanning electron microscope (Phillips FEI Co., The Netherlands).

[0315] To detect the encapsulation efficiency (EE) and drug loading (DL), 40 μL of the final composition was mixed with 160 μL of acetonitrile, followed by vortexing for 15 s and water sonication for 5 min. After centrifugation at 12,000 rpm for 8 min, the supernatant was collected for high performance liquid chromatography (HPLC) measurement. EE and DL were calculated by the following formula:

[0316] EE(%)=ma-mb / ma×100%

[0317] DL(%)=ma-mb / (ma+100mg)×100%

[0318] Wherein, ma represents the amount of active agent (eg, nucleic acid) in the initial chloroform solution, and mb represents the content of active agent in lipid nanoparticles and liposomes detected by HPLC.

[0319] Example 3

[0320] In vitro and in vivo transfection, pharmacokinetics and efficacy

[0321] In vitro and in vivo transfection can be detected in cells or mice. In order to study the pharmacokinetics of the preparation, wild-type C57BL / 6 mice (male, 8 weeks) were randomly assigned to different groups and administered at 5min, 0.5h, 4h, 8h, 24h and 48h after administration, blood was collected and plasma was separated after centrifugation (13,300rpm, 10min). Subsequently, mice were euthanized, and major organs including liver, spleen, lung, lymph node, fat pad and tumor were harvested, weighed and homogenized. The content of active agent in tissue homogenate and plasma was detected by liquid chromatography-mass spectrometry (LC-MS), using ABI-5500Qtrap (Sciex, Ontario, Canada) mass spectrometer, which has an electrospray ionization source and is connected to a Shimadzu high performance liquid chromatography (HPLC) system with an Xbridge C18 column (50×2.1mm ID, 3.5μm). The efficacy of the preparation was evaluated on different disease models.

[0322] It should be understood that the foregoing detailed description and the accompanying examples are merely illustrative and should not be taken as limiting the scope of the present disclosure, which is defined solely by the appended claims and their equivalents.

[0323] Various changes and modifications to the disclosed embodiment will become apparent to those skilled in the art and can be made without departing from the spirit and scope thereof.

Claims

1. A compound of formula (I) or a pharmaceutically acceptable salt thereof, wherein: R 1 is hydrogen or C1-C6 alkyl; X is selected from O, NR 2 , S and key; R 2 is hydrogen, C1-C6 alkyl, C2-C6 alkenyl, C2-C6 alkynyl, C3-C6 cycloalkyl or halo-C1-C6 alkyl; and L is selected from 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.

2. The compound according to claim 1 or a pharmaceutically acceptable salt thereof, wherein R 1 It's methyl.

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

4. The compound or pharmaceutically acceptable salt thereof according to any one of claims 1 to 3, wherein L has formula (A): in: 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; and R a and R b Each independently selected from C6-C 40 Alkyl, C6-C 40 Alkenyl, C6-C 40 Heteroalkyl and C6-C 40 Heteroalkenyl.

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. The compound or pharmaceutically acceptable salt thereof according to any one of claims 1 to 5, wherein L has formula (D), (E) or (F): in: 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; and R a1 and R a2 Each independently selected from C6-C 40 Alkyl and C6-C 40 Alkenyl.

7. The compound of 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. The compound or pharmaceutically acceptable salt thereof according to any one of claims 1 to 7, wherein L is selected from:

9. The compound of claim 1, wherein the compound is selected from the group consisting of:

10. A lipid composition comprising the compound according to any one of claims 1 to 9.

11. A lipid composition as claimed in claim 10, further comprising one or more cations and / or ionizable lipids, phospholipids, neutral or non-cationic lipids or combinations thereof.

12. The lipid composition of claim 10 or 11, further comprising a structured lipid.

13. A lipid composition as claimed in claim 12, wherein the structural lipid is selected from the group consisting of cholesterol, coprosterol, sitosterol, ergosterol, campesterol, stigmasterol, brassicerol, tomatine, cholesterol hemisuccinate, cholesterol sulfate, ursolic acid, alpha-tocopherol and mixtures thereof.

14. The lipid composition of any one of claims 10 to 13, further comprising a polyethylene glycol (PEG)-lipid conjugate.

15. The lipid composition of any one of claims 10 to 14, further comprising at least one activating agent.

16. lipid composition as claimed in claim 15, wherein the at least one activating agent is encapsulated by liposomes, lipid nanoparticles, or micelles in the lipid composition.

17. The lipid composition of claim 15 or 16, wherein the at least one active agent comprises a nucleic acid or a protein.

18. A lipid composition as described in any one of claims 15 to 17, wherein the at least one activating agent comprises RNA.

19. A lipid composition as described in claim 18, wherein 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 mixture thereof.

20. The lipid composition of any one of claims 17 to 19, wherein the nucleic acid encodes an antigen or a functional fragment thereof.

21. The lipid composition of any one of claims 10 to 20, wherein the at least one activating agent comprises an immunomodulator, a chemotherapeutic, a steroid, an analgesic, an antiseptic or a combination thereof.

22. A pharmaceutical composition comprising a lipid composition as described in any one of claims 10 to 21 and a pharmaceutically acceptable carrier.

23. A vaccine comprising a lipid composition as described in any one of claims 10 to 21 and an antigen or a nucleic acid encoding the same.

24. The vaccine of claim 23, wherein the antigen is a tumor antigen, an autoantigen, or an infectious disease-derived antigen.

25. The vaccine of claim 23 or 24, wherein the nucleic acid is messenger RNA (mRNA).

26. A method of delivering an active agent to a cell, comprising contacting the cell with a lipid composition as described in any one of claims 10 to 21 or a pharmaceutical composition as described in claim 22.

27. A method of delivering an active agent to a subject, comprising administering to the subject a lipid composition as described in any one of claims 10 to 21 or a pharmaceutical composition as described in claim 22.

28. A method for producing an interested polypeptide in a cell, the method comprising contacting the cell with a lipid composition as described in any one of claims 10 to 21 or a pharmaceutical composition as described in claim 22, wherein the lipid composition comprises an mRNA encoding the interested polypeptide.

29. The method of claim 28, wherein the cell is in a subject and the contacting comprises administering to the subject.

30. A method for treating or preventing a disease or condition in a subject, comprising administering to the subject in need thereof an effective amount of a lipid composition as described in any one of claims 10 to 21, a pharmaceutical composition as described in claim 22, or a vaccine as described in any one of claims 23 to 25.

31. The method of claim 30, wherein the disease or condition comprises cancer, an autoimmune disease, an inflammatory disease, or an infectious disease.

32. The method of claim 30 or 31, wherein the disease or condition is cancer.

33. The method of claim 31 or 32, wherein the subject has cancer, has had cancer, is susceptible to cancer, or has a family history of cancer.

34. The method of any one of claims 31 to 33, wherein the cancer comprises a solid tumor or a hematological cancer.

35. The method of any one of claims 31 to 34, wherein the cancer is a metastatic cancer.

36. The method of any one of claims 31 to 35, wherein the method inhibits or eliminates cancer metastasis, reduces tumor growth, prevents tumor recurrence, or any combination thereof.

37. The method of any one of claims 30 to 36, wherein the administration comprises an initial administration and at least one subsequent administration.

38. The method of any one of claims 30 to 37, wherein the subject is a human.

39. The method of any one of claims 30 to 38, further comprising administering at least one additional active agent.

40. The method of claim 38, wherein the at least one additional active agent comprises an immunomodulator, a chemotherapeutic agent, a nucleic acid, a steroid, an analgesic, an antibacterial agent, or a combination thereof.

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