JP2023520047A - Phenolic acid lipid cationic lipid - Google Patents

Abstract

The present invention, in part, provides phenolic acid lipid compounds of formula (I) and subformulae thereof, or pharmaceutically acceptable salts thereof. The compounds provided herein may be useful, for example, as components of liposomal delivery vehicles, for the delivery and expression of mRNA and encoded proteins, and thus those associated with deficiency of one or more proteins. can be useful in the treatment of various diseases, disorders, and conditions such as [Chemical 1] TIFF2023520047000258.tif43105 [Selection] Figure 1

Description

CROSS-REFERENCE TO RELATED APPLICATIONS This application claims priority to U.S. Provisional Patent Application No. 63/003,698, filed April 1, 2020, which is incorporated by reference in its entirety. .

Nucleic acid delivery has been extensively investigated as a potential therapeutic option for certain medical conditions. Specifically, messenger RNA (mRNA) therapy has become an increasingly important option for the treatment of various diseases, including those associated with deficiencies of one or more proteins.

Efficient delivery of liposome-encapsulated nucleic acids remains an active research area. The cationic lipid component plays an important role in facilitating effective encapsulation of nucleic acids during liposome loading. In addition, cationic lipids may play an important role in the efficient release of nucleic acid cargo from liposomes into the cytoplasm of target cells. A variety of cationic lipids have been found suitable for use in vivo. However, there remains a need to identify lipids that can be synthesized efficiently and inexpensively without forming potentially toxic by-products.

Phenolic acids have many advantageous features that make them good starting points for the synthesis of cationic lipids for use in an in vivo setting. For example, phenolic acids are non-toxic, available in large amounts, and can be easily derivatized. Generally, phenolic acids can be divided into two groups, benzoic acid and cinnamic acid, and their derivatives.

Examples of benzoic acids that can be used to synthesize the cationic lipids of the invention include:

Examples of cinnamic acids that can be used to synthesize the cationic lipids of the invention include:

Examples of cinnamic acids that, in some embodiments, can be used to synthesize the cationic lipids of the present invention include:

Examples of cinnamic acids that, in some embodiments, can be used to synthesize the cationic lipids of the present invention include:

Examples of cinnamic acids that, in some embodiments, can be used to synthesize the cationic lipids of the present invention include:

Examples of cinnamic acids that, in some embodiments, can be used to synthesize the cationic lipids of the present invention include:

The present invention inter alia provides novel cationic lipid compounds for the in vivo delivery of therapeutic agents, such as nucleic acids. It is contemplated that these compounds are capable of highly effective in vivo delivery while maintaining a favorable toxicity profile.

The cationic lipids of the invention can be synthesized from readily available starting reagents such as phenolic acid, benzoic acid, and cinnamic acid. The cationic lipids of the invention also have unexpectedly high encapsulation efficiencies. The cationic lipids of the invention also contain cleavable groups (eg, esters and disulfides) that are intended to improve biodegradability and thus contribute to their favorable toxicity profile.

式中、Ｌ_１は結合、（Ｃ_１－Ｃ_６）アルキルまたは（Ｃ_２－Ｃ_６）アルケニルであり、
式中、ＸはＯまたはＳであり、
式中、Ｒ^１、Ｒ^２、Ｒ^３、Ｒ^４およびＲ^５は、それぞれ独立してＨ、ＯＨ、任意で置換された（Ｃ_１－Ｃ_６）アルキル、任意で置換された（Ｃ_２－Ｃ_６）アルケニル、任意で置換された（Ｃ_２－Ｃ_６）アルキニル、任意で置換された（Ｃ_１－Ｃ_６）アルコキシおよび－ＯＣ（Ｏ）Ｒ’から選択され、
式中、Ｒ^１、Ｒ^２、Ｒ^３、Ｒ^４またはＲ^５の少なくとも一つは、－ＯＣ（Ｏ）Ｒ’であり、
式中、Ｒ’は以下であり、

式中、Ｒ^６は以下であり、

式中、ｍおよびｐはそれぞれ独立して、０、１、２、３、４、または５であり、
式中、Ｒ^７は、Ｈ、任意で置換された（Ｃ_１－Ｃ_６）アルキル、任意で置換された（Ｃ_２－Ｃ_６）アルケニル、任意で置換された（Ｃ_２－Ｃ_６）アルキニル、任意で置換された（Ｃ_１－Ｃ_６）アシル、－（ＣＨ_２）_ｋＲ^Ａまたは－（ＣＨ_２）_ｋＣＨ（ＯＲ^１１）Ｒ^Ａから選択され、
式中、Ｒ^８は、Ｈ、任意で置換された（Ｃ_１－Ｃ_６）アルキル、任意で置換された（Ｃ_２－Ｃ_６）アルケニル、任意で置換された（Ｃ_２－Ｃ_６）アルキニル、任意で置換された（Ｃ_１－Ｃ_６）アシル、－（ＣＨ_２）_ｎＲ^Ｂまたは－（ＣＨ_２）_ｎＣＨ（ＯＲ^１２）Ｒ^Ｂから選択され、
式中、Ｒ^９は、Ｈ、任意で置換された（Ｃ_１－Ｃ_６）アルキル、任意で置換された（Ｃ_２－Ｃ_６）アルケニル、任意で置換された（Ｃ_２－Ｃ_６）アルキニル、任意で置換された（Ｃ_１－Ｃ_６）アシル、－（ＣＨ_２）_ｑＲ^Ｃまたは－（ＣＨ_２）_ｑＣＨ（ＯＲ^１３）Ｒ^Ｃから選択され、
式中、Ｒ^１０は、Ｈ、任意で置換された（Ｃ_１－Ｃ_６）アルキル、任意で置換された（Ｃ_２－Ｃ_６）アルケニル、任意で置換された（Ｃ_２－Ｃ_６）アルキニル、任意で置換された（Ｃ_１－Ｃ_６）アシル、－（ＣＨ_２）_ｒＲ^Ｄまたは－（ＣＨ_２）_ｒＣＨ（ＯＲ^１４）Ｒ^Ｄから選択され、
式中、ｋ、ｎ、ｑおよびｒはそれぞれ独立して１、２、３、４、または５であり、
または式中、（ｉ）Ｒ^７およびＲ^８、または（ｉｉ）Ｒ^９およびＲ^１０は共に、任意で置換された５員または６員ヘテロシクロアルキルまたはヘテロアリールを形成し、ヘテロシクロアルキルまたはヘテロアリールは、Ｎ、ＯおよびＳから選択される１～３個のヘテロ原子を含み、
式中、Ｒ^１１、Ｒ^１２、Ｒ^１３、およびＲ^１４はそれぞれ独立してＨ、メチル、エチル、またはプロピルから選択され、
式中、Ｒ^Ａ、Ｒ^Ｂ、Ｒ^ＣおよびＲ^Ｄはそれぞれ独立して、任意で置換された（Ｃ_６－Ｃ_２０）アルキル、任意で置換された（Ｃ_６－Ｃ_２０）アルケニル、任意で置換された（Ｃ_６－Ｃ_２０）アルキニル、任意で置換された（Ｃ_６－Ｃ_２０）アシル、任意で置換された－ＯＣ（Ｏ）アルキル、任意で置換された－ＯＣ（Ｏ）アルケニル、任意で置換された（Ｃ_１－Ｃ_６）モノアルキルアミノ、任意で置換された（Ｃ_１－Ｃ_６）ジアルキルアミノ、任意で置換された（Ｃ_１－Ｃ_６）アルコキシ、－ＯＨ、－ＮＨ_２から選択され、
式中、Ｒ^７、Ｒ^８、Ｒ^９、Ｒ^１０のうちの少なくとも一つは、それぞれＲ^Ａ、Ｒ^Ｂ、Ｒ^ＣまたはＲ^Ｄ部分を含み、Ｒ^Ａ、Ｒ^Ｂ、Ｒ^ＣまたはＲ^Ｄは独立して、任意で置換された（Ｃ_６－Ｃ_２０）アルキル、任意で置換された（Ｃ_６－Ｃ_２０）アルケニル、任意で置換された（Ｃ_６－Ｃ_２０）アルキニル、任意で置換された（Ｃ_６－Ｃ_２０）アシル、任意で置換された－ＯＣ（Ｏ）（Ｃ_６－Ｃ_２０）アルキルまたは任意で置換された－ＯＣ（Ｏ）（Ｃ_６－Ｃ_２０）アルケニルから選択されるカチオン性脂質、
またはその薬学的に許容可能な塩が提供される。 In one aspect, as described herein, a cationic lipid having a structure according to formula (I):

wherein L ₁ is a bond, (C ₁ -C ₆ )alkyl or (C ₂ -C ₆ )alkenyl;
wherein X is O or S;
wherein R ¹ , R ² , R ³ , R ⁴ and R ⁵ are each independently H, OH, optionally substituted (C ₁ -C ₆ )alkyl, optionally substituted (C ₂ - C ₆ )alkenyl, optionally substituted (C ₂ -C ₆ )alkynyl, optionally substituted (C ₁ -C ₆ )alkoxy and —OC(O)R′;
wherein at least one of R ¹ , R ² , R ³ , R ⁴ or R ⁵ is —OC(O)R′;
wherein R' is

wherein ^R6 is

wherein m and p are each independently 0, 1, 2, 3, 4, or 5;
wherein R ⁷ is H, optionally substituted (C ₁ -C ₆ )alkyl, optionally substituted (C ₂ -C ₆ )alkenyl, optionally substituted (C ₂ -C ₆ )alkynyl , optionally substituted (C ₁ -C ₆ )acyl, —(CH ₂ ) _k R ^A or —(CH ₂ ) _k CH(OR ¹¹ )R ^A ;
wherein R ⁸ is H, optionally substituted (C ₁ -C ₆ )alkyl, optionally substituted (C ₂ -C ₆ )alkenyl, optionally substituted (C ₂ -C ₆ )alkynyl , optionally substituted (C ₁ -C ₆ )acyl, —(CH ₂ ) _n R ^B or —(CH ₂ ) _n CH(OR ¹² )R ^B ;
wherein R ⁹ is H, optionally substituted (C ₁ -C ₆ )alkyl, optionally substituted (C ₂ -C ₆ )alkenyl, optionally substituted (C ₂ -C ₆ )alkynyl , optionally substituted (C ₁ -C ₆ )acyl, —(CH ₂ ) _q R ^C or —(CH ₂ ) _q CH(OR ¹³ )R ^C ;
wherein R ¹⁰ is H, optionally substituted (C ₁ -C ₆ )alkyl, optionally substituted (C ₂ -C ₆ )alkenyl, optionally substituted (C ₂ -C ₆ )alkynyl , optionally substituted (C ₁ -C ₆ )acyl, —(CH ₂ ) _r R ^D or —(CH ₂ ) _r CH(OR ¹⁴ )R ^D ;
wherein k, n, q and r are each independently 1, 2, 3, 4, or 5;
or wherein (i) R ⁷ and R ⁸ or (ii) R ⁹ and R ¹⁰ together form an optionally substituted 5- or 6-membered heterocycloalkyl or heteroaryl, heterocycloalkyl or heterocycloalkyl aryl contains 1 to 3 heteroatoms selected from N, O and S;
wherein R ¹¹ , R ¹² , R ¹³ and R ¹⁴ are each independently selected from H, methyl, ethyl or propyl;
wherein R ^A , R ^B , R ^C and R ^D are each independently optionally substituted (C ₆ -C ₂₀ )alkyl, optionally substituted (C ₆ -C ₂₀ )alkenyl, optionally substituted (C ₆ -C ₂₀ )alkynyl, optionally substituted (C ₆ -C ₂₀ )acyl, optionally substituted —OC(O)alkyl, optionally substituted —OC(O)alkenyl, optionally substituted (C ₁ -C ₆ )monoalkylamino, optionally substituted (C ₁ -C ₆ )dialkylamino, optionally substituted (C ₁ -C ₆ )alkoxy, —OH, —NH selected from ₂ ,
wherein at least one of R ⁷ , R ⁸ , R ⁹ , R ¹⁰ each comprises a R ^A , R ^B , R ^C or R ^D moiety and R ^A , R ^B , R ^C or R ^D is Independently optionally substituted (C ₆ -C ₂₀ )alkyl, optionally substituted (C ₆ -C ₂₀ )alkenyl, optionally substituted (C ₆ -C ₂₀ )alkynyl, optionally substituted (C ₆ -C ₂₀ )acyl, optionally substituted —OC(O)(C ₆ -C ₂₀ )alkyl or optionally substituted —OC(O)(C ₆ -C ₂₀ )alkenyl a cationic lipid that
or a pharmaceutically acceptable salt thereof.

In one aspect, provided herein are cationic lipids that are pharmaceutically acceptable salts of Formula (I).

In one aspect, provided herein are compositions comprising a cationic lipid of the invention, one or more non-cationic lipids, one or more cholesterol-based lipids, and one or more PEG-modified lipids. be. In one aspect, the composition is a lipid nanoparticle, optionally a liposome.

In one aspect, compositions comprising the cationic lipids of the invention can be used in therapy.

FIG. 1 shows in vivo protein expression following intratracheal administration of lipid nanoparticles containing one of cationic lipid compounds 1-12. Lipid nanoparticles containing cationic lipids described herein are effective for delivery of FFL mRNA in vivo based on positive luciferase activity.

DEFINITIONS To make the invention easier to understand, certain terms are first defined below. Additional definitions for the following terms and other terms are found throughout the specification. The publications and other references referred to herein to explain the background of the invention and to provide additional details regarding its practice are hereby incorporated by reference.

Amino Acid: As used herein, the term "amino acid" in its broadest sense refers to any compound and/or substance that can be incorporated into a polypeptide chain. In some embodiments, the amino acid has the general structure H ₂ N--C(H)(R)--COOH. In some embodiments, the amino acid is a naturally occurring amino acid. In some embodiments the amino acids are synthetic amino acids, in some embodiments the amino acids are D-amino acids, and in some embodiments the amino acids are L-amino acids. "Standard amino acid" refers to any of the twenty standard L-amino acids commonly found in naturally occurring peptides. "Nonstandard amino acid" refers to any amino acid, other than the standard amino acids, regardless of whether it is prepared synthetically or obtained from a natural source. As used herein, "synthetic amino acid" encompasses chemically modified amino acids including, but not limited to, salts, amino acid derivatives (such as amides), and/or substitutions. Amino acids, including the carboxy-terminal amino acid and/or the amino-terminal amino acid in the peptide, may be methylated, amidated, acetylated, protected groups, and/or altered in the circulating half-life of the peptide without adversely affecting their activity. can be modified by substitution with other chemical groups capable of Amino acids can participate in disulfide bonds. An amino acid is one or more chemical entities (e.g., methyl group, acetate group, acetyl group, phosphate group, formyl moiety, isoprenoid group, sulfate group, polyethylene glycol moiety, lipid moiety, carbohydrate moiety, biotin moiety, etc.). may include one or post-translational modifications such as association with The term "amino acid" is used interchangeably with "amino acid residue" and can refer to free amino acids and/or amino acid residues of peptides. Whether the term refers to a free amino acid or to residues of a peptide will become clear from the context in which it is used.

Animal: As used herein, the term "animal" refers to any member of the animal kingdom. In some embodiments, "animal" refers to humans, at any stage of development. In some embodiments, "animal" refers to non-human animals, at any stage of development. In certain embodiments, non-human animals are mammals (eg, rodents, mice, rats, rabbits, monkeys, dogs, cats, sheep, cows, primates, and/or pigs). In some embodiments, animals include, but are not limited to, mammals, birds, reptiles, amphibians, fish, insects, and/or parasites. In some embodiments, animals can be transgenic animals, genetically engineered animals, and/or clones.

Approximately or about: As used herein, the terms “about” or “about” as applied to one or more values of interest refer to values similar to the stated reference value. In certain embodiments, the term "approximately" or "about" is used to refer to a given reference, unless otherwise indicated or clear from context (unless such number exceeds 100% of the possible values). 25%, 20%, 19%, 18%, 17%, 16%, 15%, 14%, 13%, 12%, 11%, 10% in either direction of the value , 9%, 8%, 7%, 6%, 5%, 4%, 3%, 2%, 1%, or less.

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

Delivery: As used herein, the term "delivery" encompasses both local and systemic delivery. For example, delivery of mRNA can include situations in which the mRNA is delivered to the target tissue, the encoded protein is expressed and retained in the target tissue (also referred to as "local distribution" or "local delivery"), and the mRNA is A situation in which the encoded protein is delivered to the target tissue, is expressed, is secreted into the patient's circulatory system (e.g., serum), is distributed throughout the body, and is taken up by other tissues (also "systemic distribution" or "systemic delivery"). referred to as).

Expression: As used herein, “expression” of a nucleic acid sequence includes translation of mRNA into a polypeptide, assembly of multiple polypeptides into an intact protein (e.g., enzyme), and/or Or refers to post-translational modifications of fully assembled proteins (eg, enzymes). In this application, the terms "expression" and "production" and their grammatical equivalents are used interchangeably.

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

Half-life: As used herein, the term "half-life" refers to the time required for a quantity, such as a nucleic acid or protein concentration or activity, to drop to half of its originally measured value for a period of time. is.

Helper lipid: As used herein, the term "helper lipid" refers to any naturally occurring or zwitterionic lipid material that contains cholesterol. While not wishing to be bound by any particular theory, helper lipids may provide stability, rigidity, and/or fluidity within the lipid bilayer/nanoparticle.

Improve, increase, or reduce: As used herein, “improve,” “increase,” or “decrease,” or grammatical synonyms, refer to baseline measures, e.g. Values compared to measurements in the same individual prior to initiation of treatment as described herein or in a control subject (or control subjects) in the absence of treatment as described herein are presented. A "control subject" is a subject who has the same form of disease as the subject being treated and who is about the same age as the subject being treated.

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

In vivo: As used herein, the term “in vivo” refers to events that occur within multicellular organisms, such as humans and non-human animals. In the context of cell-based systems, the term may be used to refer to events that occur within living cells (eg, as opposed to in vitro systems).

Isolated: As used herein, the term "isolated" means (1) associated when first produced (whether in the natural and/or experimental environment) Refers to a substance and/or entity that is separate from at least some of its components and/or (2) produced, prepared, and/or manufactured by man. Isolated substances and/or entities are about 10%, about 20%, about 30%, about 40%, about 50%, about 60%, about 70% of the other components with which they were originally associated , about 80%, about 90%, about 91%, about 92%, about 93%, about 94%, about 95%, about 96%, about 97%, about 98%, about 99%, or more than about 99% can be separated from In some embodiments, the isolated agent is about 80%, about 85%, about 90%, about 91%, about 92%, about 93%, about 94%, about 95%, about 96%, About 97%, about 98%, about 99%, or greater than about 99% pure. As used herein, a substance is "pure" if it is substantially free of other ingredients. As used herein, percent purity calculations for isolated substances and/or entities should not include excipients (eg, buffers, solvents, water, etc.).

Liposome: As used herein, the term "liposome" refers to any lamellar, multilamellar, or solid nanoparticulate vesicle. Typically, liposomes as used herein can be formed by mixing one or more lipids or by mixing one or more lipids and polymers. In some embodiments, liposomes suitable for the present invention comprise cationic and optionally non-cationic lipids, optionally cholesterol-based lipids, and/or optionally PEG-modified lipids.

Messenger RNA (mRNA): As used herein, the term “messenger RNA (mRNA)” or “mRNA” refers to a polynucleotide that encodes at least one polypeptide. As used herein, mRNA includes both modified and unmodified RNA. The term "modified mRNA" relates to mRNA containing at least one chemically modified nucleotide. An mRNA may contain one or more coding and non-coding regions. mRNA can be purified from natural sources, produced using recombinant expression systems, and optionally purified, chemically synthesized, and the like. Optionally, for example, in the case of chemically synthesized molecules, mRNA can include nucleoside analogs, such as analogs with chemically modified bases or sugars, backbone modifications, and the like. mRNA sequences are presented in the 5' to 3' direction unless otherwise indicated. In some embodiments, the mRNA contains natural nucleosides (eg, adenosine, guanosine, cytidine, uridine), nucleoside analogs (eg, 2-aminoadenosine, 2-thiothymidine, inosine, pyrrolo-pyrimidine, 3-methyladenosine, 5-methylcytidine, C-5 propynyl-cytidine, C-5 propynyl-uridine, 2-aminoadenosine, C5-bromouridine, C5-fluorouridine, C5-iodouridine, C5-propynyl-uridine, C5-propynyl-cytidine , C5-methylcytidine, 2-aminoadenosine, 7-deazaadenosine, 7-deazaguanosine, 8-oxoadenosine, 8-oxoguanosine, O(6)-methylguanine, 2-thiocytidine), chemically modified bases, biologically modified bases (e.g., methylated bases), intercalated bases, modified sugars (e.g., 2'-fluororibose, ribose, 2'-deoxyribose, arabinose, and hexose ), and/or modified phosphate groups (eg, phosphorothioate and 5′-N-phosphoramidite linkages).

Nucleic acid: As used herein, the term “nucleic acid,” in its broadest sense, refers to any compound and/or substance that is or can be incorporated into a polynucleotide chain. In some embodiments, a nucleic acid is a compound and/or substance that is or can be incorporated into a polynucleotide chain via a phosphodiester bond. In some embodiments, "nucleic acid" refers to individual nucleic acid residues (eg, nucleotides and/or nucleosides). In some embodiments, "nucleic acid" refers to a polynucleotide chain comprising individual nucleic acid residues. In some embodiments, "nucleic acid" encompasses RNA, and single- and/or double-stranded DNA and/or cDNA. In some embodiments, the "nucleic acid" includes interfering RNA (RNAi), small interfering RNA (siRNA), short hairpin RNA (shRNA), antisense RNA (aRNA), messenger RNA (mRNA), modified messenger RNA ( mmRNA), long non-coding RNAs (lncRNAs), microRNAs (miRNAs), multimer-coding nucleic acids (MCNA), polymer-coding nucleic acids (PCNA), guide RNAs (gRNAs), and CRISPR RNAs (crRNAs). Ribonucleic acid (RNA), including but not limited to one or more. In some embodiments, "nucleic acid" includes any one or more of single-stranded DNA (ssDNA), double-stranded DNA (dsDNA), and complementary DNA (cDNA), including It includes but is not limited to deoxyribonucleic acid (DNA). In some embodiments, "nucleic acid" includes both RNA and DNA. In embodiments, the DNA is antisense DNA, plasmid DNA, portions of plasmid DNA, pre-condensed DNA, polymerase chain reaction (PCR) products, vectors (e.g., P1, PAC, BAC, YAC). , artificial chromosomes), expression cassettes, chimeric sequences, chromosomal DNA, or derivatives of these groups. In embodiments, the RNA is messenger RNA (mRNA), ribosomal RNA (rRNA), signal recognition particle RNA (7 SL RNA or SRP RNA), transfer RNA (tRNA), transfer-messenger RNA (tmRNA), small molecule nuclear RNA (snRNA), small nucleolar RNA (snoRNA), SmY RNA, small cajal body-specific RNA (scaRNA), guide RNA (gRNA), ribonuclease P (RNase P), Y RNA, telomerase RNA components ( TERC), splice leader RNA (SL RNA), antisense RNA (aRNA or asRNA), cis-natural antisense transcript (cis-NAT), CRISPR RNA (crRNA), long non-coding RNA (lncRNA), microRNA ( miRNA), Piwi-binding RNA (piRNA), small interfering RNA (siRNA), trans-acting siRNA (tasiRNA), repeat-associated siRNA (rasiRNA), 73K RNA, retrotransposons, viral genomes, viroids, satellite RNAs, or any of these It can be in the form of derivatives of the group. In some embodiments, the nucleic acid is mRNA encoding a protein, such as an enzyme.

Patient: As used herein, the term "patient" or "subject" means that provided compositions can be administered, e.g., for experimental, diagnostic, prophylactic, cosmetic, and/or therapeutic purposes. means any living organism. Typical patients include animals (eg, mice, rats, rabbits, non-human primates, and/or mammals such as humans). In some embodiments, the patient is human. Human includes prenatal and postnatal forms.

Pharmaceutically acceptable: The term "pharmaceutically acceptable" as used herein means that, within the scope of good medical judgment, undue toxicity, irritation, allergic response, or other problems or substances suitable for use in contact with human and animal tissue without complications and commensurate with a reasonable benefit/risk ratio.

Pharmaceutically acceptable salts: Pharmaceutically acceptable salts are well known in the art. For example, S. M. Berge et al., for pharmaceutically acceptable salts, J. Am. described in detail in Pharmaceutical Sciences (1977) 66:1-19. Pharmaceutically acceptable salts of the compounds of this invention include those derived from suitable inorganic and organic acids and bases. Examples of pharmaceutically acceptable non-toxic acid addition salts are with inorganic acids such as hydrochloric, hydrobromic, phosphoric, sulfuric, and perchloric acids, or with acetic, oxalic, maleic, tartaric acids. , citric acid, succinic acid, or malonic acid, or by using other methods used in the art such as ion exchange. Other pharmaceutically acceptable salts include adipate, alginate, ascorbate, aspartate, benzenesulfonate, benzoate, bisulfate, borate, butyrate, camphorate. , camphorsulfonate, citrate, cyclopentanepropionate, digluconate, dodecyl sulfate, ethanesulfonate, formate, fumarate, glucoheptonate, glycerophosphate, gluconate, hemisulfate Salt, heptanoate, hexanoate, hydroiodide, 2-hydroxy-ethanesulfonate, lactobionate, lactate, laurate, lauryl sulfate, malate, maleate, malonic acid salt, methanesulfonate, 2-naphthalenesulfonate, nicotinate, nitrate, oleate, oxalate, palmitate, pamoate, pectate, persulfate, 3-phenylpropionate , phosphate, picrate, pivalate, propionate, stearate, succinate, sulfate, tartrate, thiocyanate, p-toluenesulfonate, undecanoate, valerate, etc. is mentioned. Salts derived from appropriate bases include alkali metal, alkaline earth metal, ammonium and N ⁺ (C _1-4 alkyl) ₄ salts. Representative alkali or alkaline earth metal salts include sodium, lithium, potassium, calcium, magnesium, and the like. Additional pharmaceutically acceptable salts, where appropriate, employ counterions such as halides, hydroxides, carboxylates, sulfates, phosphates, nitrates, sulfonates, and arylsulfonates. non-toxic ammonium cations, quaternary ammonium cations, and amine cations formed by Additional pharmaceutically acceptable salts include salts formed from quaternization of amines using a suitable electrophile, such as an alkyl halide, to form a quaternized alkylated amino salt. .

Systemic distribution or delivery: As used herein, the terms "systemic distribution" or "systemic delivery" or their grammatical equivalents refer to delivery or distribution mechanisms or approaches that affect the whole body or organism. . Typically, systemic distribution or delivery is accomplished via the body's circulatory system, eg, the bloodstream. Compare to the definition of "local distribution or delivery."

Subject: As used herein, the term "subject" refers to a human or any non-human animal (e.g., mouse, rat, rabbit, dog, cat, cow, pig, sheep, horse, or primate) point to Human includes prenatal and postnatal forms. In many embodiments, the subject is human. A subject can be a patient and refers to a human who sees a health care provider for the diagnosis or treatment of disease. The term "subject" is used interchangeably herein with "individual" or "patient." A subject can have or is susceptible to a disease or disorder and may or may not exhibit symptoms of the disease or disorder.

Substantially: As used herein, the term “substantially” refers to the qualitative state of exhibiting all or nearly all the extent or degree of a desired characteristic or property. Biological and chemical phenomena rarely, if ever, reach and/or reach completion, or achieve or avoid absolute results, to those skilled in the art of biology. you will understand. Thus, the term "substantially" is used herein to capture the potential lack of perfection inherent in many biological and chemical phenomena.

Target tissue: As used herein, the term "target tissue" refers to any tissue affected by the disease being treated. In some embodiments, target tissue includes tissue exhibiting a pathology, symptom, or characteristic associated with a disease.

Therapeutically Effective Amount: As used herein, the term “therapeutically effective amount” of a therapeutic agent is , means an amount sufficient to treat, diagnose, prevent, and/or delay the onset of symptoms of the disease, disorder, and/or condition. Those skilled in the art will appreciate that a therapeutically effective amount is typically administered by a dosing regimen comprising at least one unit dose.

Treating: As used herein, the terms “treat,” “treatment,” or “treating” refer to one or more symptoms or symptoms of a particular disease, disorder, and/or condition. used to partially or completely ameliorate, ameliorate, alleviate, inhibit, prevent, delay onset of, reduce severity of, and/or reduce incidence of a characteristic Refers to any method. Treatment may be given to a subject who is not exhibiting symptoms of the disease and/or who is exhibiting only early signs of the disease, with the goal of reducing the risk of developing pathology associated with the disease.

Chemical Definition Acyl: As used herein, the term “acyl” refers to R ^Z —(C═O)—, where R ^Z is, for example, any alkyl, alkenyl, alkynyl, hetero alkyl or heteroalkylene.

Aliphatic: As used herein, the term aliphatic refers to C _1- C ₄₀ hydrocarbons and includes both saturated and unsaturated hydrocarbons. Aliphatics can be linear, branched, or cyclic. For example, C ₁ -C ₂₀ aliphatic includes C ₁ -C ₂₀ alkyl (eg, linear or branched C ₁ -C ₂₀ saturated alkyl), C ₂ -C ₂₀ alkenyl (eg, linear or branched linear or branched C ₄ -C ₂₀ dienyls, linear or branched C ₆ -C ₂₀ trienyls, etc.), and C ₂ -C ₂₀ alkynyls (eg linear or branched C ₂ -C ₂₀ alkynyls). . C ₁ -C ₂₀ aliphatics can include C ₃ -C ₂₀ cycloaliphatic (eg, C ₃ -C ₂₀ cycloalkyl, C ₄ -C ₂₀ cycloalkenyl, or C ₈ -C ₂₀ cycloalkynyl) . In certain embodiments, aliphatic may contain one or more cycloaliphatics and/or one or more heteroatoms such as oxygen, nitrogen, or sulfur, optionally alkyl, halo, It can be substituted with one or more substituents such as alkoxyl, hydroxy, amino, aryl, ether, ester, or amide. Aliphatic groups are unsubstituted or substituted with one or more substituents described herein. For example, aliphatic includes halogen, -COR'', -CO ₂ H, -CO ₂ R'', -CN, -OH, -OR'', -OCOR', -OCO ₂ R'', -NH ₂ , —NHR″, —N(R″) ₂ , —SR″, or —SO ₂ R″ (eg, 1, 2, 3, 4, 5, or 6 independently selected substituents), wherein each instance of R″ is independently C ₁ -C ₂₀ aliphatic (eg, C ₁ -C ₂₀ alkyl, C ₁ -C ₁₅ alkyl, C ₁ -C ₁₀ alkyl, or C ₁ -C ₃ alkyl). In embodiments, R'' is independently unsubstituted alkyl (eg, unsubstituted C ₁ -C ₂₀ alkyl, C ₁ -C ₁₅ alkyl, C ₁ -C ₁₀ alkyl, or C ₁ -C ₃ alkyl ). In embodiments, R'' is independently unsubstituted C ₁ -C ₃ alkyl. In embodiments, aliphatics are unsubstituted. In some embodiments, aliphatic does not contain any heteroatoms. Alkyl: As used herein, the term “alkyl” refers to non-cyclic straight and branched hydrocarbon groups, for example, “C ₁ -C ₃₀ alkyl” refers to 1-30 Refers to an alkyl group containing carbon. Alkyl groups can be straight or branched. Examples of alkyl groups include, but are not limited to, methyl, ethyl, n-propyl, isopropyl, butyl, isobutyl, sec-butyl, tert-butyl, pentyl, isopentyl, tert-pentylhexyl, isohexyl, and the like. The term “lower alkyl” means straight or branched alkyl of alkyl groups having 1 to 6 carbon atoms. Other alkyl groups will be readily apparent to those skilled in the art given the benefit of this disclosure. An alkyl group can be unsubstituted or substituted with one or more substituents described herein. For example, alkyl groups include halogen, -COR'', -CO ₂ H, -CO ₂ R'', -CN, -OH, -OR'', -OCOR', -OCO ₂ R'', -NH ₂ , —NHR″, —N(R″) ₂ , —SR″, or —SO ₂ R″ (eg, 1, 2, 3, 4, 5, or 6 independently selected substituents), wherein each instance of R″ is independently C ₁ -C ₂₀ aliphatic (eg, C ₁ -C ₂₀ alkyl, C ₁ -C ₁₅ alkyl, C ₁ -C ₁₀ alkyl, or C ₁ -C ₃ alkyl). In embodiments, R'' is independently unsubstituted alkyl (eg, unsubstituted C ₁ -C ₂₀ alkyl, C ₁ -C ₁₅ alkyl, C ₁ -C ₁₀ alkyl, or C ₁ -C ₃ alkyl ). In embodiments, R'' is independently unsubstituted C ₁ -C ₃ alkyl. In embodiments, alkyl is substituted (eg, with 1, 2, 3, 4, 5, or 6 substituents described herein). In embodiments, alkyl groups are substituted with -OH groups, which may also be referred to herein as "hydroxyalkyl" groups, where the prefix indicates a -OH group and "alkyl" is herein as described in

As used herein, “alkyl” also refers to the radical of a straight or branched chain saturated hydrocarbon group having from 1 to 50 carbon atoms (“C ₁ -C ₅₀ alkyl”). . In some embodiments, alkyl groups have from 1 to 40 carbon atoms (“C ₁ -C ₄₀ alkyl”). In some embodiments, an alkyl group has from 1 to 30 carbon atoms (“C ₁ -C ₃₀ alkyl”). In some embodiments, alkyl groups have from 1 to 20 carbon atoms (“C ₁ -C ₂₀ alkyl”). In some embodiments, an alkyl group has from 1 to 10 carbon atoms (“C ₁ -C ₁₀ alkyl”). In some embodiments, alkyl groups have from 1 to 9 carbon atoms (“C ₁ -C ₉ alkyl”). In some embodiments, the alkyl group has 1-8 carbon atoms (“C ₁ -C ₈ alkyl”). In some embodiments, an alkyl group has from 1 to 7 carbon atoms (“C ₁ -C ₇ alkyl”). In some embodiments, an alkyl group has from 1 to 6 carbon atoms (“C ₁ -C ₆ alkyl”). In some embodiments, an alkyl group has 1-5 carbon atoms (“C ₁ -C ₅ alkyl”). In some embodiments, an alkyl group has 1 to 4 carbon atoms (“C ₁ -C ₄ alkyl”). In some embodiments, an alkyl group has 1 to 3 carbon atoms (“C ₁ -C ₃ alkyl”). In some embodiments, an alkyl group has 1-2 carbon atoms (“C ₁ -C ₂ alkyl”). In some embodiments, an alkyl group has 1 carbon atom (“C ₁ alkyl”). In some embodiments, an alkyl group has from 2 to 6 carbon atoms (“C ₂ -C ₆ alkyl”). Examples of C ₁ -C ₆ alkyl groups include, but are not limited to, methyl (C ₁ ), ethyl (C ₂ ), n-propyl (C ₃ ), isopropyl (C _{3 )} , n-butyl ( C ₄ ), tert-butyl (C ₄ ), sec-butyl (C ₄ ), iso-butyl (C ₄ ), n-pentyl (C ₅ ), 3-pentanyl (C ₅ ), amyl (C ₅ ), Included are neopentyl (C ₅ ), 3-methyl-2-butanyl (C ₅ ), tertiary amyl (C ₅ ), and n-hexyl (C ₆ ). Additional examples of alkyl groups include n-heptyl (C ₇ ), n-octyl (C ₈ ), and the like. Unless otherwise specified, each instance of an alkyl group is independently unsubstituted (“unsubstituted alkyl”) or substituted with one or more substituents (“substituted alkyl”). In certain embodiments, alkyl groups are unsubstituted C ₁ -C ₅₀ alkyl. In certain embodiments, alkyl groups are substituted C ₁ -C ₅₀ alkyl.

If a radical is suffixed with "-ene", it becomes a divalent moiety. For example, arylene is the divalent moiety of aryl and heteroarylene is the divalent moiety of heteroaryl.

Alkylene: The term "alkylene," as used herein, represents a saturated divalent straight or branched chain hydrocarbon radical, exemplified by methylene, ethylene, isopropylene, and the like. Similarly, the term "alkenylene" as used herein refers to an unsaturated divalent linear chain having one or more unsaturated carbon-carbon double bonds that may occur at any stable point along the chain. or branched chain hydrocarbon group, the term "alkynylene" as used herein refers to an unsaturated divalent hydrocarbon group having one or more unsaturated carbon-carbon triple bonds that may occur at any stable point along the chain. Represents a straight or branched chain hydrocarbon group. In certain embodiments, an alkylene, alkenylene, or alkynylene group can contain one or more cycloaliphatic and/or one or more heteroatoms such as oxygen, nitrogen, or sulfur, alkyl, halo, can be optionally substituted with one or more substituents such as , alkoxyl, hydroxy, amino, aryl, ether, ester, or amide. For example, alkylene, alkenylene, or alkynylene is halogen, -COR'', -CO ₂ H, -CO ₂ R'', -CN, -OH, -OR'', -OCOR'', -OCO ₂ R'', -NH ₂ , -NHR'', -N(R'') ₂ , -SR'', or -SO ₂ R'' (e.g., 1, 2, 3, 4, 5, or 6 independently selected substituents), wherein each instance of R″ is independently C ₁ -C ₂₀ aliphatic (eg, C ₁ -C ₂₀ alkyl, C ₁ -C ₁₅ alkyl, C ₁ -C ₁₀ alkyl, or C ₁ -C ₃ alkyl). In embodiments, R'' is independently unsubstituted alkyl (eg, unsubstituted C ₁ -C ₂₀ alkyl, C ₁ -C ₁₅ alkyl, C ₁ -C ₁₀ alkyl, or C ₁ -C ₃ alkyl ). In embodiments, R'' is independently unsubstituted C ₁ -C ₃ alkyl. In certain embodiments, an alkylene, alkenylene, or alkynylene is unsubstituted. In certain embodiments, an alkylene, alkenylene, or alkynylene does not contain any heteroatoms. Alkenyl: As used herein, “alkenyl” refers to any linear or straight chain having one or more unsaturated carbon-carbon double bonds that may occur at any stable point along the chain. A branched hydrocarbon chain is meant, for example, “C ₂ -C ₃₀ alkenyl” refers to alkenyl groups having 2 to 30 carbons. For example, alkenyl groups include prop-2-enyl, but-2-enyl, but-3-enyl, 2-methylprop-2-enyl, hex-2-enyl, hex-5-enyl, 2,3-dimethyl but-2-enyl and the like are included. In embodiments, alkenyl contains 1, 2, or 3 carbon-carbon double bonds. In embodiments, alkenyl contains a single carbon-carbon double bond. In embodiments, multiple double bonds (eg, two or three) are conjugated. An alkenyl group can be unsubstituted or substituted with one or more substituents described herein. For example, alkenyl groups include halogen, -COR'', -CO ₂ H, -CO ₂ R'', -CN, -OH, -OR'', -OCOR'', -OCO ₂ R'', -NH ₂ , —NHR″, —N(R″) ₂ , —SR″, or —SO ₂ R″ (eg, 1, 2, 3, 4, 5, or 6 independently selected substituents), wherein each instance of R″ is independently C ₁ -C ₂₀ aliphatic (eg, C ₁ -C ₂₀ alkyl, C ₁ -C ₁₅ alkyl, C ₁ -C ₁₀ alkyl, or C ₁ -C ₃ alkyl). In embodiments, R'' is independently unsubstituted alkyl (eg, unsubstituted C ₁ -C ₂₀ alkyl, C ₁ -C ₁₅ alkyl, C ₁ -C ₁₀ alkyl, or C ₁ -C ₃ alkyl ). In embodiments, R'' is independently unsubstituted C ₁ -C ₃ alkyl. In some embodiments, alkenyl is unsubstituted. In embodiments, the alkenyl is substituted (eg, with 1, 2, 3, 4, 5, or 6 substituents described herein). In embodiments, alkenyl groups are substituted with -OH groups, which may also be referred to herein as "hydroxyalkenyl" groups, where the prefix indicates a -OH group and "alkenyl" is herein as described in

As used herein, “alkenyl” also includes 2 to 50 carbon atoms and one or more carbon-carbon double bonds (eg, 1, 2, 3, or 4 double bonds). (“C ₂ -C ₅₀ alkenyl”) refers to the radical of a straight-chain or branched hydrocarbon group having In some embodiments, an alkenyl group has from 2 to 40 carbon atoms (“C ₂ -C ₄₀ alkenyl”). In some embodiments, an alkenyl group has from 2 to 30 carbon atoms (“C ₂ -C ₃₀ alkenyl”). In some embodiments, an alkenyl group has from 2 to 20 carbon atoms (“C ₂ -C ₂₀ alkenyl”). In some embodiments, an alkenyl group has from 2 to 10 carbon atoms (“C ₂ -C ₁₀ alkenyl”). In some embodiments, an alkenyl group has from 2 to 9 carbon atoms (“C ₂ -C ₉ alkenyl”). In some embodiments, an alkenyl group has from 2 to 8 carbon atoms (“C ₂ -C ₈ alkenyl”). In some embodiments, an alkenyl group has from 2 to 7 carbon atoms (“C ₂ -C ₇ alkenyl”). In some embodiments, an alkenyl group has from 2 to 6 carbon atoms (“C ₂ -C ₆ alkenyl”). In some embodiments, an alkenyl group has 2-5 carbon atoms (“C ₂ -C ₅ alkenyl”). In some embodiments, an alkenyl group has 2-4 carbon atoms (“C ₂ -C ₄ alkenyl”). In some embodiments, an alkenyl group has 2-3 carbon atoms (“C ₂ -C ₃ alkenyl”). In some embodiments, an alkenyl group has 2 carbon atoms (“ _C2 alkenyl”). The one or more carbon-carbon double bonds can be internal (eg, 2-butenyl) or terminal (eg, 1-butenyl). Examples of C ₂ -C ₄ alkenyl groups include, but are not limited to, ethenyl (C ₂ ), 1-propenyl (C ₃ ), 2-propenyl (C ₃ ), 1-butenyl (C ₄ ), 2-butenyl ( C ₄ ), butadienyl (C ₄ ), and the like. Examples of C ₂ -C ₆ alkenyl groups include the aforementioned C ₂ -C ₄ alkenyl groups, as well as pentenyl (C ₅ ), pentadienyl (C ₅ ), hexenyl (C6), and the like. Additional examples of alkenyl include heptenyl ( _C7 ), octenyl ( _C8 ), octatrienyl ( _C8 ), and the like. Unless specified otherwise, each instance of an alkenyl group is independently unsubstituted (“unsubstituted alkenyl”) or substituted with one or more substituents (“substituted alkenyl”). In certain embodiments, alkenyl groups are unsubstituted C ₂ -C ₅₀ alkenyls. In certain embodiments, alkenyl groups are substituted C ₂ -C ₅₀ alkenyls.

Alkynyl: As used herein, "alkynyl" is either a linear or branched arrangement having one or more carbon-carbon triple bonds occurring at any stable point along the chain. Any hydrocarbon chain is meant, for example, “C ₂ -C ₃₀ alkynyl” refers to alkynyl groups having 2 to 30 carbons. Examples of alkynyl groups include prop-2-ynyl, but-2-ynyl, but-3-ynyl, pent-2-ynyl, 3-methylpent-4-ynyl, hex-2-ynyl, hex-5-ynyl. etc. In some embodiments, alkynyl contains one carbon-carbon triple bond. An alkynyl group can be unsubstituted or substituted with one or more substituents described herein. For example, alkynyl groups include halogen, -COR'', _-CO2H , _-CO2R '', -CN, -OH, -OR'', -OCOR'', _-OCO2R '', -NH ₂ , —NHR″, —N(R″) ₂ , —SR″, or —SO ₂ R″ (eg, 1, 2, 3, 4, 5, or 6 independently selected substituents), wherein each instance of R″ is independently C ₁ -C ₂₀ aliphatic (eg, C ₁ -C ₂₀ alkyl, C ₁ -C ₁₅ alkyl, C ₁ -C ₁₀ alkyl, or C ₁ -C ₃ alkyl). In embodiments, R'' is independently unsubstituted alkyl (eg, unsubstituted C ₁ -C ₂₀ alkyl, C ₁ -C ₁₅ alkyl, C ₁ -C ₁₀ alkyl, or C ₁ -C ₃ alkyl ). In embodiments, R'' is independently unsubstituted C ₁ -C ₃ alkyl. In some embodiments, alkynyl is unsubstituted. In embodiments, alkynyl is substituted (eg, with 1, 2, 3, 4, 5, or 6 substituents described herein).

As used herein, “alkynyl” also includes 2-50 carbon atoms and one or more carbon-carbon triple bonds (eg, 1, 2, 3, or 4 triple bonds) and optionally Refers to the radical of a straight or branched hydrocarbon group having one or more double bonds (eg, 1, 2, 3, or 4 double bonds) in (“C ₂ -C ₅₀ alkynyl”). Alkynyl groups having one or more triple bonds and one or more double bonds are also referred to as "ene-yne." In some embodiments, an alkynyl group has from 2 to 40 carbon atoms (“C ₂ -C ₄₀ alkynyl”). In some embodiments, an alkynyl group has from 2 to 30 carbon atoms (“C ₂ -C ₃₀ alkynyl”). In some embodiments, an alkynyl group has from 2 to 20 carbon atoms (“C ₂ -C ₂₀ alkynyl”). In some embodiments, an alkynyl group has from 2 to 10 carbon atoms (“C ₂ -C ₁₀ alkynyl”). In some embodiments, an alkynyl group has from 2 to 9 carbon atoms (“C ₂ -C ₉ alkynyl”). In some embodiments, an alkynyl group has from 2 to 8 carbon atoms (“C ₂ -C ₈ alkynyl”). In some embodiments, an alkynyl group has from 2 to 7 carbon atoms (“C ₂ -C ₇ alkynyl”). In some embodiments, an alkynyl group has from 2 to 6 carbon atoms (“C ₂ -C ₆ alkynyl”). In some embodiments, an alkynyl group has 2-5 carbon atoms (“C ₂ -C ₅ alkynyl”). In some embodiments, an alkynyl group has 2-4 carbon atoms (“C ₂ -C ₄ alkynyl”). In some embodiments, an alkynyl group has 2-3 carbon atoms (“C ₂ -C ₃ alkynyl”). In some embodiments, an alkynyl group has 2 carbon atoms (“ _C2 alkynyl”). The one or more carbon-triple bonds can be internal (eg, 2-butynyl) or terminal (eg, 1-butynyl). Examples of C ₂ -C ₄ alkynyl groups include, but are not limited to, ethynyl (C ₂ ), 1-propynyl (C ₃ ), 2-propynyl (C ₃ ), 1-butynyl (C ₄ ), 2-butynyl ( C ₄ ) and the like. Examples of C ₂ -C ₆ alkenyl groups include the aforementioned C ₂ -C ₄ alkynyl groups, as well as pentynyl (C ₅ ), hexynyl (C ₆ ), and the like. Further examples of alkynyl include heptynyl ( _C7 ), octynyl ( _C8 ), and the like. Unless otherwise specified, each instance of an alkynyl group is independently unsubstituted (“unsubstituted alkynyl”) or substituted with one or more substituents (“substituted alkynyl”). In certain embodiments, alkynyl groups are unsubstituted C ₂ -C ₅₀ alkynyl. In certain embodiments, alkynyl groups are substituted C ₂ -C ₅₀ alkynyls.

Aryl: The term “aryl” used alone or as part of a larger moiety, as in “aralkyl,” refers to a monocyclic, bicyclic, or means a tricyclic carbocyclic ring structure, wherein said ring structure has a single point of attachment to the rest of the molecule, at least one ring in the system is aromatic, and wherein the system Each inner ring contains 4 to 7 ring members. In some embodiments, an aryl group has 6 ring carbon atoms (“ _C6 aryl”; eg, phenyl). In some embodiments, an aryl group has 10 ring carbon atoms (“C ₁₀ aryl”; eg, naphthyl such as 1-naphthyl and 2-naphthyl). In some embodiments, an aryl group has 14 ring carbon atoms (“C ₁₄ aryl”; eg, anthracyl). "Aryl" also includes ring systems in which the aryl ring is fused with one or more carbocyclyl or heterocyclyl groups, as defined above, wherein the linking radical or point of attachment is on the aryl ring, in such instances , the number of carbon atoms continues to specify the number of carbon atoms in the aryl ring system. Examples of aryl include phenyl, naphthyl, and anthracene.

As used herein, “aryl” also has 6 to 14 ring carbon atoms (“C ₆ -C ₁₄ aryl”) provided in an aromatic ring system and 0 heteroatoms; Refers to a radical of a monocyclic or polycyclic (e.g., bicyclic or tricyclic) 4n+2 aromatic ring system (e.g., having 6, 10, or 14 pi-electrons shared in a cyclic arrangement) . In some embodiments, an aryl group has 6 ring carbon atoms (“ _C6 aryl”; eg, phenyl). In some embodiments, an aryl group has 10 ring carbon atoms (“C ₁₀ aryl”; eg, naphthyl such as 1-naphthyl and 2-naphthyl). In some embodiments, an aryl group has 14 ring carbon atoms (“C ₁₄ aryl”; eg, anthracyl). "Aryl" also includes ring systems in which the aryl ring is fused with one or more carbocyclyl or heterocyclyl groups, as defined above, wherein the linking radical or point of attachment is on the aryl ring, in such instances , the number of carbon atoms continues to specify the number of carbon atoms in the aryl ring system. Unless otherwise specified, each instance of an aryl group is independently unsubstituted (“unsubstituted aryl”) or substituted with one or more substituents (“substituted aryl”). In certain embodiments, aryl groups are unsubstituted C ₆ -C ₁₄ aryl. In certain embodiments, aryl groups are substituted C ₆ -C ₁₄ aryl.

Arylene: As used herein, the term "arylene" refers to a divalent aryl group (ie, having two points of attachment to the molecule). Examples of arylenes include phenylenes (eg, unsubstituted or substituted phenylenes).

Carbocyclyl: As used herein, “carbocyclyl” or “carbocyclic” means 3 to 10 ring carbon atoms (“C ₃ -C ₁₀ carbocyclyl”) and 0 ring carbon atoms in a non-aromatic ring system. refers to the radical of a non-aromatic cyclic hydrocarbon group having a heteroatom of In some embodiments, a carbocyclyl group has 3 to 8 ring carbon atoms (“C ₃ -C ₈ carbocyclyl”). In some embodiments, a carbocyclyl group has 3 to 7 ring carbon atoms (“C ₃ -C ₇ carbocyclyl”). In some embodiments, a carbocyclyl group has 3 to 6 ring carbon atoms (“C ₃ -C ₆ carbocyclyl”). In some embodiments, a carbocyclyl group has 4 to 6 ring carbon atoms (“C ₄ -C ₆ carbocyclyl”). In some embodiments, a carbocyclyl group has 5 to 6 ring carbon atoms (“C ₅ -C ₆ carbocyclyl”). In some embodiments, a carbocyclyl group has 5 to 10 ring carbon atoms (“C ₅ -C ₁₀ carbocyclyl”). Exemplary C ₃ -C ₆ carbocyclyl groups include, but are not limited to, cyclopropyl (C ₃ ), cyclopropenyl (C ₃ ), cyclobutyl (C ₄ ), cyclobutenyl (C 4 ), cyclopentyl (C ₅ ), cyclopentenyl (C ₅ ), cyclohexyl (C ₆ ), cyclohexenyl (C ₆ ), cyclohexadienyl (C ₆ ), and the like. Exemplary C ₃ -C ₈ carbocyclyl groups include, but are not limited to, the aforementioned C ₃ -C ₆ carbocyclyl groups, as well as cycloheptyl (C ₇ ), cycloheptenyl (C ₇ ), cycloheptadienyl ( _C7 ), cycloheptatrienyl ( _C7 ), cyclooctyl ( _C8 ), cyclooctenyl ( _C8 ), bicyclo[2.2.1]heptanyl ( _C7 ), bicyclo[2.2.2]octanyl ( C ₈ ) and the like. Exemplary C ₃ -C ₁₀ carbocyclyl groups include, but are not limited to, the aforementioned C ₃ -C ₈ carbocyclyl groups, as well as cyclononyl (C ₉ ), cyclononenyl (C ₉ ), cyclodecyl (C ₁₀ ), Cyclodecenyl (C ₁₀ ), octahydro-1H-indenyl (C ₉ ), decahydronaphthalenyl (C ₁₀ ), spiro[4.5]decanyl (C ₁₀ ) and the like. As the preceding examples demonstrate, in certain embodiments, a carbocyclyl group is monocyclic (“monocyclic carbocyclyl”) or polycyclic (e.g., bicyclic (“bicyclic carbocyclyl”) or tricyclic). ring systems (containing fused, bridged, or spiro ring systems such as "tricyclic carbocyclyl"), which may be saturated, or which contain one or more carbon-carbon double bonds or It may contain a triple bond. "Carbocyclyl" also includes ring systems in which the carbocyclyl ring is fused with one or more aryl or heteroaryl groups, as defined above, wherein the point of attachment is on the carbocyclyl ring, in such instances a carbon The numbers continue to specify the number of carbon atoms in the carbocyclic ring system. Unless otherwise specified, each instance of a carbocyclyl group is independently unsubstituted (“unsubstituted carbocyclyl”) or substituted with one or more substituents (“substituted carbocyclyl”). In certain embodiments, the carbocyclyl group is unsubstituted C _3- C ₁₀ carbocyclyl. In certain _embodiments , a carbocyclyl group is a substituted C ₃ -C 10 carbocyclyl.

In some embodiments, "carbocyclyl" or "carbocyclic" refers to "cycloalkyl", i.e., a monocyclic saturated carbocyclyl group having 3 to 10 ring carbon atoms (" _C3 - _C10 ”). In some embodiments, a cycloalkyl group has from 3 to 8 ring carbon atoms (“C ₃ -C ₈ cycloalkyl”). In some embodiments, a cycloalkyl group has from 3 to 6 ring carbon atoms (“C ₃ -C ₆ cycloalkyl”). In some embodiments, a cycloalkyl group has 4 to 6 ring carbon atoms (“C ₄ -C ₆ cycloalkyl”). In some embodiments, a cycloalkyl group has 5 to 6 ring carbon atoms (“C ₅ -C ₆ cycloalkyl”). In some embodiments, a cycloalkyl group has from 5 to 10 ring carbon atoms (“C ₅ -C ₁₀ cycloalkyl”). Examples of C ₅ -C ₆ cycloalkyl groups include cyclopentyl (C ₅ ) and cyclohexyl (C ₅ ). Examples of C3- _C6 cycloalkyl groups include the aforementioned _C5 - _C6 cycloalkyl groups, as well as cyclopropyl ( _C3 ) and cyclobutyl ( _C4 ). Examples of C ₃ -C ₈ cycloalkyl groups include the aforementioned C ₃ -C ₆ cycloalkyl groups, as well as cycloheptyl (C ₇ ) and cyclooctyl (C ₈ ). Unless otherwise specified, each instance of a cycloalkyl group is independently unsubstituted (“unsubstituted cycloalkyl”) or substituted with one or more substituents (“substituted cycloalkyl”). . In certain embodiments, a cycloalkyl group is an unsubstituted C ₃ -C ₁₀ cycloalkyl. In certain embodiments, a cycloalkyl group is a substituted C ₃ -C ₁₀ cycloalkyl.

Halogen: As used herein, the term "halogen" means fluorine, chlorine, bromine, or iodine.

Heteroalkyl: The term "heteroalkyl" refers to 1 to 14 carbon atoms in addition to 1, 2, 3 or 4 heteroatoms independently selected from the group consisting of N, O, S, and P. means a branched or unbranched alkyl, alkenyl, or alkynyl group having a Heteroalkyl includes tertiary amines, secondary amines, ethers, thioethers, amides, thioamides, carbamates, thiocarbamates, hydrazones, imines, phosphodiesters, phosphoramidates, sulfonamides, and disulfides. Heteroalkyl groups may optionally include monocyclic, bicyclic, or tricyclic rings, each ring desirably having 3-6 members. Examples of heteroalkyl include polyethers such as methoxymethyl and ethoxyethyl.

Heteroalkylene: As used herein, the term "heteroalkylene" represents the divalent form of the heteroalkyl groups described herein.

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

As used herein, "heteroaryl" also includes ring carbon atoms provided in an aromatic ring system and one or more (e.g., 1, 2, 3, or 4 ring heteroatoms) 5- to 14-membered monocyclic or polycyclic (e.g., bicyclic or tricyclic) 4n+2 aromatic ring systems with heteroatoms (e.g., 6, 10, or 14 π shared in a cyclic arrangement with electrons), each heteroatom being independently selected from oxygen, sulfur, nitrogen, boron, silicon, and phosphorus (“5- to 14-membered heteroaryl”). In heteroaryl groups containing one or more nitrogen atoms, the point of attachment can be a carbon or nitrogen atom, as valences permit. Heteroaryl polycyclic ring systems can include one or more heteroatoms in one or both rings. "Heteroaryl" includes ring systems in which the heteroaryl ring is fused with one or more carbocyclyl or heterocyclyl groups, as defined above, wherein the point of attachment is on the heteroaryl ring, in such instances The number of ring members continues to specify the number of ring members in the heteroaryl ring system. "Heteroaryl" also includes ring systems in which a heteroaryl ring is fused to one or more aryl groups, as defined above, wherein the point of attachment is on either the aryl ring or the heteroaryl ring. , and in such instances, the number of ring members specifies the number of ring members in a fused polycyclic (aryl/heteroaryl) ring system. Polycyclic heteroaryl groups in which one ring does not contain a heteroatom (e.g., indolyl, quinolinyl, carbazolyl, etc.) are those in which the point of attachment is either ring, i.e., a ring containing a heteroatom (e.g., 2-indolyl). or in any heteroatom-free ring (eg, 5-indolyl).

In some embodiments, the heteroaryl group has 5 to 10 ring carbon atoms provided in the aromatic ring system and one or more (eg, 1, 2, 3, or 4) ring heteroatoms. It is a membered aromatic ring system in which each heteroatom is independently selected from oxygen, sulfur, nitrogen, boron, silicon, and phosphorus (“5- to 10-membered heteroaryl”). In some embodiments, the heteroaryl group has 5-8 ring carbon atoms provided in the aromatic ring system and one or more (eg, 1, 2, 3, or 4) ring heteroatoms. It is a membered aromatic ring system where each heteroatom is independently selected from oxygen, sulfur, nitrogen, boron, silicon, and phosphorus (“5- to 8-membered heteroaryl”). In some embodiments, the heteroaryl group has 5-6 ring carbon atoms provided in the aromatic ring system and one or more (eg, 1, 2, 3, or 4) ring heteroatoms. It is a membered aromatic ring system wherein each heteroatom is independently selected from oxygen, sulfur, nitrogen, boron, silicon, and phosphorus (“5- to 6-membered heteroaryl”). In some embodiments, the 5-6 membered heteroaryl has one or more (eg, 1, 2, or 3) ring heteroatoms selected from oxygen, sulfur, nitrogen, boron, silicon, and phosphorus. have In some embodiments, the 5-6 membered heteroaryl has 1 or 2 ring heteroatoms selected from oxygen, sulfur, nitrogen, boron, silicon, and phosphorus. In some embodiments, the 5-6 membered heteroaryl has 1 ring heteroatom selected from oxygen, sulfur, nitrogen, boron, silicon, and phosphorus. Unless otherwise specified, each instance of a heteroaryl group is independently unsubstituted (“unsubstituted heteroaryl”) or substituted with one or more substituents (“substituted heteroaryl”). . In certain embodiments, the heteroaryl group is an unsubstituted 5-14 membered heteroaryl. In certain embodiments, the heteroaryl group is a substituted 5-14 membered heteroaryl.

Exemplary 5-membered heteroaryl groups containing one heteroatom include, without limitation, pyrrolyl, furanyl, and thiophenyl. Exemplary 5-membered heteroaryl groups containing two heteroatoms include, without limitation, imidazolyl, pyrazolyl, oxazolyl, isoxazolyl, thiazolyl, and isothiazolyl. Exemplary 5-membered heteroaryl groups containing 3 heteroatoms include, without limitation, triazolyl, oxadiazolyl, and thiadiazolyl. Exemplary 5-membered heteroaryl groups containing 4 heteroatoms include, without limitation, tetrazolyl. Exemplary 6-membered heteroaryl groups containing one heteroatom include, without limitation, pyridinyl. Exemplary 6-membered heteroaryl groups containing two heteroatoms include, without limitation, pyridazinyl, pyrimidinyl, and pyrazinyl. Exemplary 6-membered heteroaryl groups containing 3 or 4 heteroatoms include, without limitation, triazinyl and tetrazinyl, respectively. Exemplary 7-membered heteroaryl groups containing one heteroatom include, without limitation, azepinyl, oxepinyl, and thiepinyl. Exemplary 5,6-bicyclic heteroaryl groups include, but are not limited to, indolyl, isoindolyl, indazolyl, benzotriazolyl, benzothiophenyl, isobenzothiophenyl, benzofuranyl, benzisofuranyl, benzimidazolyl, Benzoxazolyl, benzoxazolyl, benzoxadiazolyl, benzthiazolyl, benzisothiazolyl, benzthiadiazolyl, indolizinyl, and purinyl. Exemplary 6,6-bicyclic heteroaryl groups include, without limitation, napthyridinyl, pteridinyl, quinolinyl, isoquinolinyl, cinnolinyl, quinoxalinyl, phthalazinyl, and quinazolinyl. Exemplary tricyclic heteroaryl groups include, without limitation, phenanthridinyl, dibenzofuranyl, carbazolyl, acridinyl, phenothiazinyl, phenoxazinyl, and phenazinyl.

As used herein, “heterocyclyl” or “heterocycle” refers to 3 to 14 ring carbon atoms and one or more (eg, 1, 2, 3, or 4) ring heteroatoms. Refers to a membered non-aromatic ring system radical, where each heteroatom is independently selected from oxygen, sulfur, nitrogen, boron, silicon, and phosphorus (“3- to 14-membered heterocyclyl”). In heterocyclyl groups containing one or more nitrogen atoms, the point of attachment can be a carbon or nitrogen atom, as valences permit. A heterocyclyl group may be monocyclic (“monocyclic heterocyclyl”) or polycyclic (e.g., fused, bridged, or spiro ring systems, e.g., bicyclic (“bicyclic heterocyclyl”) or tricyclic systems (“tricyclic heterocyclyl”)) and may be saturated or may contain one or more carbon-carbon double or triple bonds. Heterocyclyl polycyclic ring systems can include one or more heteroatoms in one or both rings. "Heterocyclyl" is a ring system in which a heterocyclyl ring, as defined above, is fused with one or more carbocyclyl groups (where the point of attachment is on either the carbocyclyl ring or the heterocyclyl ring), or Also included are ring systems in which a heterocyclyl ring is fused with one or more aryl or heteroaryl groups (where the point of attachment is on the heterocyclyl ring), in which case the number of ring members is equal to the number of ring members in the heterocyclyl ring system. continue to specify the number of Unless otherwise specified, each instance of heterocyclyl is independently unsubstituted (“unsubstituted heterocyclyl”) or substituted with one or more substituents (“substituted heterocyclyl”). In certain embodiments, the heterocyclyl group is unsubstituted 3-14 membered heterocyclyl. In certain embodiments, the heterocyclyl group is a substituted 3-14 membered heterocyclyl.

In some embodiments, the heterocyclyl group is a 5-10 membered non-aromatic ring system having ring carbon atoms and one or more (eg, 1, 2, 3, or 4) ring heteroatoms; Each heteroatom is independently selected from oxygen, sulfur, nitrogen, boron, silicon, and phosphorus (“5-10 membered heterocyclyl”). In some embodiments, the heterocyclyl group is a 5-8 membered non-aromatic ring system having ring carbon atoms and one or more (eg, 1, 2, 3, or 4) ring heteroatoms; Each heteroatom is independently selected from oxygen, sulfur, nitrogen, boron, silicon, and phosphorus (“5-8 membered heterocyclyl”). In some embodiments, the heterocyclyl group is a 5-6 membered non-aromatic ring system having ring carbon atoms and one or more (eg, 1, 2, 3, or 4) ring heteroatoms; Each heteroatom is independently selected from oxygen, sulfur, nitrogen, boron, silicon, and phosphorus (“5-6 membered heterocyclyl”). In some embodiments, the 5-6 membered heterocyclyl has one or more (eg, 1, 2, or 3) ring heteroatoms selected from oxygen, sulfur, nitrogen, boron, silicon, and phosphorus. have. In some embodiments, the 5-6 membered heterocyclyl has 1 or 2 ring heteroatoms selected from oxygen, sulfur, nitrogen, boron, silicon, and phosphorus. In some embodiments, the 5-6 membered heterocyclyl has 1 ring heteroatom selected from oxygen, sulfur, nitrogen, boron, silicon, and phosphorus.

Exemplary 3-membered heterocyclyl groups containing one heteroatom include, without limitation, azirdinyl, oxiranyl, thiorenyl. Exemplary 4-membered heterocyclyl groups containing one heteroatom include, without limitation, azetidinyl, oxetanyl, and thietanyl. Exemplary 5-membered heterocyclyl groups containing one heteroatom include tetrahydrofuranyl, dihydrofuranyl, tetrahydrothiophenyl, dihydrothiophenyl, pyrrolidinyl, dihydropyrrolyl, and pyrrolyl-2,5-dione; , but not limited to. Exemplary 5-membered heterocyclyl groups containing two heteroatoms include, without limitation, dioxolanyl, oxathiolanyl, and dithiolanyl. Exemplary 5-membered heterocyclyl groups containing 3 heteroatoms include, without limitation, triazolinyl, oxadiazolinyl, and thiadiazolinyl. Exemplary 6-membered heterocyclyl groups containing one heteroatom include, without limitation, piperidinyl, tetrahydropyranyl, dihydropyridinyl, and thianyl. Exemplary 6-membered heterocyclyl groups containing two heteroatoms include, without limitation, piperazinyl, morpholinyl, dithianyl, dioxanyl. Exemplary 6-membered heterocyclyl groups containing two heteroatoms include, without limitation, triazinanyl. Exemplary 7-membered heterocyclyl groups containing one heteroatom include, without limitation, azepanyl, oxepanyl, and thiepanyl. Exemplary 8-membered heterocyclyl groups containing one heteroatom include, without limitation, azocanyl, oxecanyl, and thiocanyl. Exemplary bicyclic heterocyclyl groups include indolinyl, isoindolinyl, dihydrobenzofuranyl, dihydrobenzothienyl, tetrahydrobenzothienyl, tetrahydrobenzofuranyl, tetrahydroindolyl, tetrahydroquinolinyl, tetrahydroisoquinolinyl, decahydroxyl. nolinyl, decahydroisoquinolinyl, octahydrochromenyl, octahydroisochromenyl, decahydronaphthyridinyl, decahydro-1,8-naphthyridinyl, octahydropyrrolo[3,2-b]pyrrole, indolinyl, phthalimidyl , naphthalimidyl, chromanyl, chromenyl, 1H-benzo[e][1,4]diazepinyl, 1,4,5,7-tetrahydropyrano[3,4-b]pyrrolyl, 5,6-dihydro-4H-furo[ 3,2-b]pyrrolyl, 6,7-dihydro-5H-furo[3,2-b]pyranyl, 5,7-dihydro-4H-thieno[2,3-c]pyranyl, 2,3-dihydro- 1H-pyrrolo[2,3-b]pyridinyl, 2,3-dihydrofuro[2,3-b]pyridinyl, 4,5,6,7-tetrahydro-1H-pyrrolo-[2,3-b]pyridinyl, 4 ,5,6,7-tetrahydrofuro[3,2-c]pyridinyl, 4,5,6,7-tetrahydrothieno[3,2-b]pyridinyl, 1,2,3,4-tetrahydro-1,6 - including but not limited to naphthyridinyl and the like.

Heterocycloalkyl: As used herein, the term “heterocycloalkyl” means that at least one atom is a heteroatom such as, but not limited to, nitrogen, oxygen, sulfur, or phosphorus, and the remainder is a non-aromatic ring in which the atoms of are carbon. A heterocycloalkyl group can be substituted or unsubstituted.

As understood from the above, alkyl, alkenyl, alkynyl, acyl, carbocyclyl, heterocyclyl, aryl, and heteroaryl groups are optionally substituted in certain embodiments. Optionally substituted refers to groups that may be substituted or unsubstituted (e.g., “substituted” or “unsubstituted” alkyl groups, “substituted” or “unsubstituted” alkenyl groups, “substituted” or “unsubstituted” alkynyl group, "substituted" or "unsubstituted" heteroalkyl group, "substituted" or "unsubstituted" heteroalkenyl group, "substituted" or "unsubstituted" heteroalkynyl group, "substituted" or "unsubstituted" carbocyclyl group, "substituted" or "unsubstituted" heterocyclyl groups, "substituted" or "unsubstituted" aryl groups, or "substituted" or "unsubstituted" heteroaryl groups). In general, the term "substituted" means that at least one hydrogen present on a group is replaced by an acceptable substituent, e.g., a compound stabilized by substitution, e.g., rearrangement, cyclization, elimination, or other reactions, etc. is substituted with substituents that result in the compound not spontaneously undergoing transformations by Unless otherwise indicated, a "substituted" group has substituents at one or more substitutable positions of the group, and when more than one position in any given structure is substituted, Substituents are either the same or different at each position. The term "substituted" is contemplated to include all permissible substituents of organic compounds, substitutions with any of the substituents described herein that result in the formation of stable compounds. The present invention contemplates any and all such combinations in order to arrive at a stable compound. For purposes of this invention, heteroatoms such as nitrogen have hydrogen substituents and/or any suitable substituents described herein that satisfy the valences of the heteroatom and result in the formation of a stable moiety. can.

Exemplary carbon atom substituents include, but are not limited to, halogen, -CN, -NO2, -N3, -SO2, -SO3H, -OH, -ORaa, -ON(Rbb)2, -N (Rbb)2, -N(Rbb)3+X-, -N(ORcc)Rbb, -SeH, -SeRaa, -SH, -SRaa, -SSRcc, -C(=O)Raa, -COH, -CHO, - C(ORcc)2, -CO2Raa, -OC(=O)Raa, -OCO2Raa, -C(=O)N(Rbb)2, -OC(=O)N(Rbb)2, -NRbbC(=O) Raa, -NRbbCO2Raa, -NRbbC(=O)N(Rbb)2, -C(=NRbb)Raa, -C(=NRbb)ORaa, -OC(=NRbb)Raa, -OC(=NRbb)ORaa, - C(=NRbb)N(Rbb)2,-OC(=NRbb)N(Rbb)2,-NRbbC(=NRbb)N(Rbb)2,-C(=O)NRbbSO2Raa,-NRbbSO2Raa,-SO2N(Rbb ) 2, -SO2Raa, -SO2ORaa, -OSO2Raa, -S(=O)Raa, -OS(=O)Raa, -Si(Raa)3-OSi(Raa)3-C(=S)N(Rbb) 2, -C(=O)SRaa, -C(=S)SRaa, -SC(=S)SRaa, -SC(=O)SRaa, -OC(=O)SRaa, -SC(=O)ORaa, -SC(=O)Raa, -P(=O)2Raa, -OP(=O)2Raa, -P(=O)(Raa)2, -OP(=O)(Raa)2, -OP(= O)(ORcc)2, -P(=O)2N(Rbb)2, -OP(=O)2N(Rbb)2, -P(=O)(NRbb)2, -OP(=O)(NRbb ) 2, -NRbbP (= O) (ORcc) 2, -NRbbP (= O) (NRbb) 2, -P (Rcc) 2, -P (Rcc) 3, -OP (Rcc) 2, -OP (Rcc ) 3, -B(Raa)2, -B(ORcc)2, -BRaa(ORcc), C1-C50 alkyl, C2-C50 alkenyl, C2-C50 alkynyl, C3-C14 carbocyclyl, 3-14 membered heterocyclyl, C6 —C14 aryl, and 5- to 14-membered heteroaryl, where each alkyl, alkenyl, alkynyl, carbocyclyl, heterocyclyl, aryl, and heteroaryl is independently 0, 1, 2, 3, 4, or 5 Rdd groups is replaced by;

or two geminal hydrogens on the carbon atom are =O group, =S group, =NN(Rbb)2 group, =NNRbbC(=O)Raa group, =NNRbbC(=O)ORaa group, =NNRbbS(=O ) substituted with a 2Raa group, =NRbb group, or =NORcc group;

Each instance of Raa is independently selected from C1-C50 alkyl, C2-C50 alkenyl, C2-C50 alkynyl, C3-C10 carbocyclyl, 3-14 membered heterocyclyl, C6-C14 aryl, and 5-14 membered heteroaryl or two Raa groups are taken together to form a 3-14 membered heterocyclyl or 5-14 membered heteroaryl ring, each alkyl, alkenyl, alkynyl, carbocyclyl, heterocyclyl, aryl, and heteroaryl independently , 0, 1, 2, 3, 4, or 5 substituted with Rdd groups;

each instance of Rbb is independently hydrogen, -OH, -ORaa, -N(Rcc)2, -CN, -C(=O)Raa, -C(=O)N(Rcc)2, -CO2Raa , -SO2Raa, -C(=NRcc)ORaa, -C(=NRcc)N(Rcc)2, -SO2N(Rcc)2, -SO2Rcc, -SO2ORcc, -SORaa, -C(=S)N(Rcc) 2, -C(=O)SRcc, -C(=S)SRcc, -P(=O)2Raa, -P(=O)(Raa)2, -P(=O)2N(Rcc)2,- P(=O)(NRcc)2, selected from C1-C50 alkyl, C2-C50 alkenyl, C2-C50 alkynyl, C3-C10 carbocyclyl, 3-14 membered heterocyclyl, C6-C14 aryl, and 5-14 membered heteroaryl or two Rbb groups together with the heteroatom to which they are attached form a 3-14 membered heterocyclyl or 5-14 membered heteroaryl ring, each alkyl, alkenyl, alkynyl, carbocyclyl, heterocyclyl , aryl, and heteroaryl are independently substituted with 0, 1, 2, 3, 4, or 5 Rdd groups;

Each instance of Rcc is independently hydrogen, C1-C50 alkyl, C2-C50 alkenyl, C2-C50 alkynyl, C3-C10 carbocyclyl, 3-14 membered heterocyclyl, C6-C14 aryl, and 5-14 membered heteroaryl or two Rcc groups taken together with the heteroatom to which they are attached form a 3-14 membered heterocyclyl or 5-14 membered heteroaryl ring, each alkyl, alkenyl, alkynyl, carbocyclyl , heterocyclyl, aryl, and heteroaryl are independently substituted with 0, 1, 2, 3, 4, or 5 Rdd groups;

Each instance of Rdd is independently halogen, -CN, -NO2, -N3, -SO2H, -SO3H, -OH, -ORee, -ON(Rff)2, -N(Rff)2, -N( Rff) 3+X-, -N(ORee)Rff, -SH, -SRee, -SSRee, -C(=O)Ree, -CO2H, -CO2Ree, -OC(=O)Ree, -OCO2Ree, -C(= O)N(Rff)2,-OC(=O)N(Rff)2,-NRffC(=O)Ree,-NRffCO2Ree,-NRffC(=O)N(Rff)2,-C(=NRff)ORee , -OC (=NRff) Ree, -OC (=NRff) ORee, -C (=NRff) N (Rff) 2, -OC (=NRff) N (Rff) 2, -NRffC (=NRff) N (Rff ) 2, -NRffSO2Ree, -SO2N (Rff) 2, -SO2Ree, -SO2ORee, -OSO2Ree, -S (=O) Ree, -Si (Ree) 3, -OSi (Ree) 3, -C (=S) N(Rff)2, -C(=O)SRee, -C(=S)SRee, -SC(=S)SRee, -P(=O)2Ree, -P(=O)(Ree)2, - OP(=O)(Ree)2, -OP(=O)(ORee)2, C1-C50 alkyl, C2-C50 alkenyl, C2-C50 alkynyl, C3-C10 carbocyclyl, 3-10 membered heterocyclyl, C6-C10 aryl, 5- to 10-membered heteroaryl, each alkyl, alkenyl, alkynyl, carbocyclyl, heterocyclyl, aryl, and heteroaryl independently substituted with 0, 1, 2, 3, 4, or 5 Rgg groups; , or two geminal Rdd substituents can be taken together to form =O or =S;

each instance of Ree is independently selected from C1-C50 alkyl, C2-C50 alkenyl, C2-C50 alkynyl, C3-C10 carbocyclyl, C6-C10 aryl, 3-10 membered heterocyclyl, and 3-10 membered heteroaryl and each alkyl, alkenyl, alkynyl, carbocyclyl, heterocyclyl, aryl, and heteroaryl is independently substituted with 0, 1, 2, 3, 4, or 5 Rgg groups;

Each instance of Rff is independently hydrogen, C1-C50 alkyl, C2-C50 alkenyl, C2-C50 alkynyl, C3-C10 carbocyclyl, 3-10 membered heterocyclyl, C6-C10 aryl, and 5-10 membered heteroaryl or two Rff groups together with the heteroatom to which they are attached form a 3-14 membered heterocyclyl or 5-14 membered heteroaryl ring, each alkyl, alkenyl, alkynyl, carbocyclyl , heterocyclyl, aryl, and heteroaryl are independently substituted with 0, 1, 2, 3, 4, or 5 Rgg groups; and

Each instance of Rgg is independently halogen, —CN, —NO2, —N3, —SO2H, —SO3H, —OH, —OC1-C50alkyl, —ON(C1-C50alkyl)2, —N(C1 —C50 alkyl) 2, —N(C1-C50 alkyl)3+X-, —NH(C1-C50 alkyl)2+X-, —NH2(C1-C50 alkyl)+X-, —NH3+X-, —N(OC1-C50 alkyl) ) (C1-C50 alkyl), —N(OH)(C1-C50 alkyl), —NH(OH), —SH, —SC1-C50 alkyl, —SS(C1-C50 alkyl), —C(═O) (C1-C50 alkyl), -CO2H, -CO2 (C1-C50 alkyl), -OC(=O) (C1-C50 alkyl), -OCO2 (C1-C50 alkyl), -C(=O)NH2, - C(=O)N(C1-C50 alkyl)2, -OC(=O)NH(C1-C50 alkyl), -NHC(=O)(C1-C50 alkyl), -N(C1-C50 alkyl)C (=O) (C1-C50 alkyl), -NHCO2 (C1-C50 alkyl), -NHC(=O) N(C1-C50 alkyl)2, -NHC(=O) NH(C1-C50 alkyl), - NHC(=O)NH, -C(=NH)O(C1-C50 alkyl), -OC(=NH)(C1-C50 alkyl), -OC(=NH)OC1-C50 alkyl, -C(=NH ) N (C1-C50 alkyl) 2, —C (=NH) NH (C1-C50 alkyl), —C (=NH) NH, —OC (=NH) N (C1-C50 alkyl) 2, —OC ( NH)NH(C1-C50 alkyl), -OC(NH)NH2, -NHC(NH)N(C1-C50 alkyl)2, -NHC(=NH)NH2, -NHSO2(C1-C50 alkyl), -SO2N (C1-C50 alkyl) 2, -SO2NH(C1-C50 alkyl), -SO2NH2, -SO2(C1-C50 alkyl), -SO2O(C1-C50 alkyl), -OSO2(C1-C6 alkyl), -SO( C1-C6 alkyl), -Si(C1-C50 alkyl)3, -OSi(C1-C6 alkyl)3, -C(=S)N(C1-C50alkyl)2, C(=S)NH(C1- C50 alkyl), C(=S)NH2, -C(=O)S(C1-C6 alkyl), -C(=S)S(C1-C6 alkyl), -SC(=S)S(C1-C6 alkyl), -P(=O)2(C1-C50 alkyl), -P(=O)(C1-C50 alkyl)2, -OP(=O)(C1-C50 alkyl)2, -OP(=O ) (OC1-C50 alkyl) 2, C1-C50 alkyl, C2-C50 alkenyl, C2-C50 alkynyl, C3-C10 carbocyclyl, C6-C10 aryl, 3-10 membered heterocyclyl, 5-10 membered heteroaryl; Or two geminal Rgg substituents can be taken together to form =O or =S; X- is a counterion.

As used herein, the term “halo” or “halogen” refers to fluorine (fluoro, —F), chlorine (chloro, —Cl), bromine (bromo, —Br), or iodine (iodo, —I ).

As used herein, a "counterion" is a negatively charged group attached to a positively charged quaternary amine to maintain electronic neutrality. Exemplary counterions include halide ions (eg, F-, Cl-, Br-, I-), NO3-, ClO4-, OH-, H2PO4-, HSO4-, sulfonate ions (eg, methanesulfone acid, trifluoromethanesulfonate, p-toluenesulfonate, benzenesulfonate, 10-camphorsulfonate, naphthalene-2-sulfonate, naphthalene-l-sulfonic acid-5-sulfonate, ethane -1-sulfonic acid-2-sulfonate, etc.) and carboxylic acid ions (e.g., acetate, ethanoate, propanoate, benzoate, glycerate, lactate, tartrate, glycolate, etc.) is mentioned.

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

In certain embodiments, substituents present on nitrogen atoms are nitrogen protecting groups (also called amino protecting groups). Nitrogen protecting groups are well known in the art and are described in Protecting Groups in Organic Synthesis, T.; W. Greene andP. G. M. Wuts, 3rd edition, John Wiley & Sons, 1999.

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

Nitrogen protecting groups such as carbamate groups (e.g., -C(=O)ORaa) include, but are not limited to, methyl carbamate, ethyl carbamante, 9-fluorenylmethyl carbamate (Fmoc), 9 -(2-sulfo)fluorenylmethylcarbamate, 9-(2,7-dibromo)fluorenylmethylcarbamate, 2,7-di-t-butyl-[9-(10,10-dioxo-10,10 ,10,10-tetrahydrothioxanthyl]]methylcarbamate (DBD-Tmoc), 4-methoxyphenacylcarbamate (Phenoc), 2,2,2-trichloroethylcarbamate (Troc), 2-trimethylsilylethylcarbamate (Teoc) , 2-phenylethylcarbamate (hZ), 1-(1-adamanty1)-1-methylethylcarbamate (Adpoc), 1,1-dimethyl-2-haloethylcarbamate, 1,1-dimethyl-2, 2-dibromoethylcarbamate (DB-t-BOC), 1,1-dimethyl-2,2,2-trichloroethylcarbamate (TCBOC), 1-methyl-1-(4-biphenylyl)ethylcarbamate (Bpoc), 1-(3,5-di-t-butylphenyl)-1-methylethylcarbamate (t-Bumeoc), 2-(2′- and 4′-pyridyl)ethylcarbamate (Pyoc), 2-(N,N -dicyclohexylcarboxamido) ethyl carbamate, t-butyl carbamate (BOC), 1-adamantyl carbamate (Adoc), vinyl carbamate (Voc), allyl carbamate (Alloc), 1-isopropyl allyl carbamate (Ipaoc), cinnamyl carbamate (Coc) , 4-nitrocinnamylcarbamate (Noc), 8-quinolylcarbamate, N-hydroxypiperidinylcarbamate, alkyldithiocarbamate, benzylcarbamate (Cbz), p-methoxybenzylcarbamate (Moz), p-nitobenzylcarbamate, p-bromobenzylcarbamate, p-chlorobenzylcarbamate, 2,4-dichlorobenzylcarbamate, 4-methylsulfinylbenzylcarbamate (Msz), 9-anthrylmethylcarbamate, diphenylmethylcarbamate, 2-methylthioethylcarbamate, 2-methyl sulfonyl ethyl carbamate, 2-(p-toluenesulfonyl) ethyl carbamate, [2-(1,3-dithianyl)] methyl carbamate (Dmoc), 4-methylthiophenyl carbamate (Mtpc), 2,4-dimethylthiophenyl carbamate ( Bmpc), 2-phosphonioethylcarbamate (Peoc), 2-triphenylphosphonioisopropylcarbamate (Ppoc), 1,1-dimethyl-2-cyanoethylcarbamate, m-chloro-p-acyloxybenzylcarbamate, p-(dihydroxy Boryl) benzyl carbamate, 5-benzisoxazolyl methyl carbamate, 2-(trifluoromethyl)-6-chromonyl methyl carbamate (Tcroc), m-nitrophenyl carbamate, 3,5-dimethoxybenzyl carbamate, o-nitro benzyl carbamate, 3,4-dimethoxy-6-nitrobenzyl carbamate, phenyl (o-nitrophenyl) methyl carbamate, t-amyl carbamate, S-benzylthiocarbamate, p-cyanobenzyl carbamate, cyclobutyl carbamate, cyclohexyl carbamate, cyclopentyl carbamate, cyclopropylmethylcarbamate, p-decyloxybenzylcarbamate, 2,2-dimethoxyacylvinylcarbamate, o-(N,N-dimethylcarboxamido)benzylcarbamate, 1,1-dimethyl-3-(N,N-dimethyl carboxamido) propyl carbamate, 1,1-dimethylpropynyl carbamate, di(2-pyridyl) methyl carbamate, 2-furanyl methyl carbamate, 2-iodoethyl carbamate, isoborynl carbamate, isobutyl carbamate, isonicotinyl carbamate, p -(p'-methoxyphenylazo)benzylcarbamate, 1-methylcyclobutylcarbamate, 1-methylcyclohexylcarbamate, 1-methyl-l-cyclopropylmethylcarbamate, 1-methyl-1(3,5-dimethoxyphenyl)ethyl carbamate, 1-methyl-1-(p-phenylazophenyl)ethylcarbamate, 1-methyl-l-phenylethylcarbamate, 1-methyl-1-(4-pyridyl)ethylcarbamate, phenylcarbamate, p-(phenylazo) benzylcarbamate, 2,4,6-tri-t-butylphenylcarbamate, 4-(trimethylammonium)benzylcarbamate, and 2,4,6-trimethylbenzylcarbamate.

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

Other nitrogen protecting groups include, but are not limited to, phenothiazinyl-(10)-acyl derivatives, N'-p-toluenesulfonylaminoacyl derivatives, N'-phenylaminothioacyl derivatives, N-benzoylphenylaryl Nil derivatives, N-acetylmethionine derivatives, 4,5-diphenyl-3-oxazolin-2-one, N-phthalimide, N-dithiasuccinimide (Dts), N-2,3-diphenylmaleimide, N-2,5 -dimethylpyrrole, N-1,1,4,4-tetramethyldisilylazacyclopentane adduct (STABASE), 5-substituted 1,3-dimethyl-1,3,5-triazacyclohex-2-one , 5-substituted 1,3-dibenzyl-1,3,5-triazacyclohexan-2-one, 1-substituted 3,5-dinitro-4-pyridone, N-methylamine, N-allylamine, N-[2 -(trimethylsilyl)ethoxy]methylamine (SEM), N-3-acetoxypropylamine, N-(1-isopropyl-4-nitro-2-oxo-3-pyroolin-3-yl)amine, quaternary ammonium salt, N-benzylamine, N-di(4-methoxyphenyl)methylamine, N-5-dibenzosuberylamine, N-triphenylmethylamine (Tr), N-[(4-methoxyphenyl)diphenylmethyl] amine (MMTr), N-9-phenylfluorenylamine (PhF), N-2,7-dichloro-9-fluorenylmethyleneamine, N-ferrocenylmethylamino (Fcm), N-2-picolylamino N '-oxide, N-1,1-dimethylthiomethyleneamine, N-benzylideneamine, Np-methoxybenzylideneamine, N-diphenylmethyleneamine, N-[(2-pyridyl)mesityl]methyleneamine, N -(N',N'-dimethylaminomethylene)amine, N,N'-isopropylidenediamine, Np-nitrobenzylideneamine, N-salicylideneamine, N-5-chlorosalicylideneamine, N- (5-chloro-2-hydroxyphenyl)phenylmethyleneamine, N-cyclohexylideneamine, N-(5,5-dimethyl-3-oxo-l-cyclohexenyl)amine, N-borane derivatives, N-diphenylborine Acid derivatives, N-[phenyl(pentacylchromium- or tungsten)acyl]amines, N-copper chelates, N-zinc chelates, N-nitroamines, N-nitrosamines, amine N-oxides, diphenylphosphine amides (Dpp), dimethyl Thiophosphinamide (Mpt), diphenylthiophosphineamide (Ppt), dialkyl phosphoramidate, dibenzyl phosphoramidate, diphenyl phosphoramidate, benzenesulfenamide, o-nitrobenzenesulfenamide (Nps), 2,4-dinitrobenzenesulfenamide pentachlorobenzenesulfenamide, 2-nitro-4-methoxybenzenesulfenamide, triphenylmethylsulfenamide, and 3-nitropyridinesulfenamide (Npys).

In certain embodiments, substituents present on oxygen atoms are oxygen protecting groups (also called hydroxyl protecting groups). Oxygen protecting groups are well known in the art and are described in Protecting Groups in Organic Synthesis, T.; W. Greene andP. G. M. Wuts, 3rd edition, John Wiley & Sons, 1999.

Exemplary oxygen protecting groups include, but are not limited to, methyl, methoxylmethyl (MOM), methylthiomethyl (MTM), t-butylthiomethyl, (phenyldimethylsilyl)methoxymethyl (SMOM), benzyloxy methyl (BOM), p-methoxybenzyloxymethyl (PMBM), (4-methoxyphenoxy)methyl (p-AOM), guaiacolmethyl (GUM), t-butoxymethyl, 4-pentenyloxymethyl (POM), siloxymethyl , 2-methoxyethoxymethyl (MEM), 2,2,2-trichloroethoxymethyl, bis(2-chloroethoxy)methyl, 2-(trimethylsilyl)ethoxymethyl (SEMOR), tetrahydropyranyl (THP), 3-bromo tetrahydropyranyl, tetrahydrothiopyranyl, 1-methoxycyclohexyl, 4-methoxytetrahydropyranyl (MTHP), 4-methoxytetrahydrothiopyranyl, 4-methoxytetrahydrothiopyranyl S,S-dioxide, 1-[( 2-chloro-4-methyl)phenyl]-4-methoxypiperidin-4-yl (CTMP), 1,4-dioxan-2-yl, tetrahydrofuranyl, tetrahydrothiofuranyl, 2,3,3a,4,5 ,6,7,7a-octahydro-7,8,8-trimethyl-4,7-methanobenzofuran-2-yl, 1-ethoxyethyl, 1-(2-chloroethoxy)ethyl, 1-methyl-l-methoxy ethyl, 1-methyl-1-benzyloxyethyl, 1-methyl-1-benzyloxy-2-fluoroethyl, 2,2,2-trichloroethyl, 2-trimethylsilylethyl, 2-(phenylselenyl)ethyl, t -butyl, allyl, p-chlorophenyl, p-methoxyphenyl, 2,4-dinitrophenyl, benzyl (Bn), p-methoxybenzyl, 3,4-dimethoxybenzyl, o-nitrobenzyl, p-nitrobenzyl, p- Halobenzyl, 2,6-dichlorobenzyl, p-cyanobenzyl, p-phenylbenzyl, 2-picolyl, 4-picolyl, 3-methyl-2-picolyl N-oxide, diphenylmethyl, p,p'-dinitrobenzhydryl , 5-dibenzosuberyl, triphenylmethyl, α-naphthyldiphenylmethyl, p-methoxyphenyldiphenylmethyl, di(p-methoxyphenyl)phenylmethyl, tri(p-methoxyphenyl)methyl, 4-(4′-bromo phenacyloxyphenyl)diphenylmethyl, 4,4′,4″-tris(4,5-dichlorophthalimidophenyl)methyl, 4,4′,4″-tris(levulinoyloxyphenyl)methyl, 4,4′ , 4″-tris(benzoyloxyphenyl)methyl, 3-(imidazol-1-yl)bis(4′,4″-dimethoxyphenyl)methyl, 1,1-bis(4-methoxyphenyl)-1′-pyrenylmethyl , 9-anthryl, 9-(9-phenyl)xanthenyl, 9-(9-phenyl-10-oxo)anthryl, 1,3-benzodisulfuran-2-yl, benzisothiazolyl S,S-dioxide, trimethylsilyl (TMS), triethylsilyl (TES), triisopropylsilyl (TIPS), dimethylisopropylsilyl (IPDMS), diethylisopropylsilyl (DEIPS), dimethylthexylsilyl, t-butyldimethylsilyl (TBDMS), t-butyldiphenylsilyl (TBDPS), tribenzylsilyl, tri-p-xylylsilyl, triphenylsilyl, diphenylmethylsilyl (DPMS), t-butylmethoxyphenylsilyl (TBMPS), formate, benzoyl formate, acetate, chloroacetate, dichloroacetate, trichloroacetate, trifluoroacetate, methoxyacetate, triphenylmethoxyacetate, phenoxyacetate, p-chlorophenoxyacetate, 3-phenylpropionate, 4-oxopentanoate (levulinate), 4,4-(ethylenedithio) Pentanoate (levulinoyldithioacetal), pivaloate, adamantate, crotonate, 4-methoxycrotonate, benzoate, p-phenylbenzoate, 2,4,6-trimethylbenzoate (mesitoate), alkylmethyl carbonate, 9-fur olenyl methyl carbonate (Fmoc), alkyl ethyl carbonate, alkyl 2,2,2-trichloroethyl carbonate (Troc), 2-(trimethylsilyl) ethyl carbonate (TMSEC), 2-(phenylsulfonyl) ethyl carbonate (Psec), 2 -(triphenylphosphonio)ethyl carbonate (Peoc), alkyl isobutyl carbonate, alkyl vinyl carbonate alkyl allyl carbonate, alkyl p-nitrophenyl carbonate, alkyl benzyl carbonate, alkyl p-methoxybenzyl carbonate, alkyl 3,4-dimethoxybenzyl carbonate , alkyl o-nitrobenzyl carbonate, alkyl p-nitrobenzyl carbonate, alkyl S-benzylthiocarbonate, 4-ethoxy-1-naptthyl carbonate, methyldithiocarbonate, 2-iodobenzoate, 4-azidobutyrate, 4 -nitro-4-methylpentanoate, o-(dibromomethyl)benzoate, 2-formylbenzenesulfonate, 2-(methylthiomethoxy)ethyl, 4-(methylthiomethoxy)butyrate, 2-(methylthiomethoxymethyl)benzoate, 2 ,6-dichloro-4-methylphenoxyacetate, 2,6-dichloro-4-(1,1,3,3-tetramethylbutyl)phenoxyacetate, 2,4-bis(1,1-dimethylpropyl)phenoxyacetate , chlorodiphenylacetate, isobutyrate, monosuccinoate, (E)-2-methyl-2-butenoate, o-(methoxyacyl)benzoate, α-naphthoate, nitrate, alkyl N,N,N',N'-tetra Methyl Phosphorodiamidates, Alkyl N-Phenyl Carbamates, Borate Salts, Dimethylphosphinothiols, Alkyl 2,4-Dinitrophenyl Sulfonates, Sulfates, Methanesulfonates (Mesylates), Benzyl Sulfonates, and Tosylates (Ts) are mentioned.

In certain embodiments, substituents present on sulfur atoms are sulfur protecting groups (also called thiol protecting groups). Sulfur protecting groups are well known in the art and are described in Protecting Groups in Organic Synthesis, T.; W. Greene andP. G. M. Wuts, 3rd edition, John Wiley & Sons, 1999.

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

Compounds of the Invention Liposomal-based vehicles are considered attractive carriers for therapeutic agents and are still the subject of ongoing development efforts. Although liposome-based vehicles containing cationic moieties have shown promising results in terms of encapsulation, stability, and site localization, there remains a great need for improved liposome-based delivery systems. For example, a significant drawback of liposome delivery systems is the construction of liposomes with sufficient cell culture or in vivo stability to reach the desired target cells and/or subcellular compartments, and the use of their encapsulated materials in such targeting. It relates to the ability of such liposomal delivery systems to efficiently release into cells.

Specifically, there remains a need for improved lipid compounds that exhibit improved pharmacokinetic properties and are capable of delivering macromolecules such as nucleic acids to a wide variety of cell types and tissues with high efficiency. Importantly, there also remains a particular need for novel lipid compounds that are characterized by reduced toxicity and are capable of efficiently delivering encapsulated nucleic acids and polynucleotides to target cells, tissues, and organs.

Described herein are novel cationic lipid compounds intended for improved in vivo delivery of therapeutic agents, such as nucleic acids. In particular, the cationic lipids described herein are optionally used with other lipids to form lipid-based nano-particles for the encapsulation of therapeutic agents, such as nucleic acids (e.g., DNA, siRNA, mRNA, microRNA) for therapeutic use. Particles (eg, liposomes) may be prepared.

In embodiments, the inventive compounds described herein may provide one or more desired characteristics or properties. That is, in certain embodiments, the compounds of the invention described herein are characterized as having one or more properties that provide advantages of such compounds compared to other similarly classified lipids. can be For example, the compounds disclosed herein can allow control and modulation of the properties of their component liposomal compositions (eg, lipid nanoparticles). Specifically, the compounds disclosed herein can be characterized by their ability to produce enhanced transfection efficiencies and specific biological outcomes. Such results may include, for example, enhanced cellular uptake, endosome/lysosome disruption capacity, and/or enhanced release of encapsulated materials (eg, polynucleotides) within cells. Additionally, the compounds described herein possess advantageous pharmacokinetic performance, biodistribution and efficacy (eg, due to different dissociation rates of the polymer groups used).

The present application demonstrates that the cationic lipids of the invention are not only synthetically transportable from readily available starting materials, but also have unexpectedly high encapsulation efficiencies.

Additionally, the cationic lipids of the present invention have cleavable groups such as ester groups and disulfides. These cleavable groups, such as esters and disulfides, are intended to improve biodegradability and thus contribute to their favorable toxicity profile.

式中、Ｌ_１は結合、（Ｃ_１－Ｃ_６）アルキルまたは（Ｃ_２－Ｃ_６）アルケニルであり、
式中、ＸはＯまたはＳであり、
式中、Ｒ^１、Ｒ^２、Ｒ^３、Ｒ^４およびＲ^５は、それぞれ独立してＨ、ＯＨ、任意で置換された（Ｃ_１－Ｃ_６）アルキル、任意で置換された（Ｃ_２－Ｃ_６）アルケニル、任意で置換された（Ｃ_２－Ｃ_６）アルキニル、任意で置換された（Ｃ_１－Ｃ_６）アルコキシおよび－ＯＣ（Ｏ）Ｒ’から選択され、
式中、Ｒ^１、Ｒ^２、Ｒ^３、Ｒ^４またはＲ^５の少なくとも一つは、－ＯＣ（Ｏ）Ｒ’であり、
式中、Ｒ’は以下であり、

式中、Ｒ^６は以下であり、

式中、ｍおよびｐはそれぞれ独立して、０、１、２、３、４、または５であり、
式中、Ｒ^７は、Ｈ、任意で置換された（Ｃ_１－Ｃ_６）アルキル、任意で置換された（Ｃ_２－Ｃ_６）アルケニル、任意で置換された（Ｃ_２－Ｃ_６）アルキニル、任意で置換された（Ｃ_１－Ｃ_６）アシル、－（ＣＨ_２）_ｋＲ^Ａまたは－（ＣＨ_２）_ｋＣＨ（ＯＲ^１１）Ｒ^Ａから選択され、
式中、Ｒ^８は、Ｈ、任意で置換された（Ｃ_１－Ｃ_６）アルキル、任意で置換された（Ｃ_２－Ｃ_６）アルケニル、任意で置換された（Ｃ_２－Ｃ_６）アルキニル、任意で置換された（Ｃ_１－Ｃ_６）アシル、－（ＣＨ_２）_ｎＲ^Ｂまたは－（ＣＨ_２）_ｎＣＨ（ＯＲ^１２）Ｒ^Ｂから選択され、
式中、Ｒ^９は、Ｈ、任意で置換された（Ｃ_１－Ｃ_６）アルキル、任意で置換された（Ｃ_２－Ｃ_６）アルケニル、任意で置換された（Ｃ_２－Ｃ_６）アルキニル、任意で置換された（Ｃ_１－Ｃ_６）アシル、－（ＣＨ_２）_ｑＲ^Ｃまたは－（ＣＨ_２）_ｑＣＨ（ＯＲ^１３）Ｒ^Ｃから選択され、
式中、Ｒ^１０は、Ｈ、任意で置換された（Ｃ_１－Ｃ_６）アルキル、任意で置換された（Ｃ_２－Ｃ_６）アルケニル、任意で置換された（Ｃ_２－Ｃ_６）アルキニル、任意で置換された（Ｃ_１－Ｃ_６）アシル、－（ＣＨ_２）_ｒＲ^Ｄまたは－（ＣＨ_２）_ｒＣＨ（ＯＲ^１４）Ｒ^Ｄから選択され、
式中、ｋ、ｎ、ｑおよびｒはそれぞれ独立して１、２、３、４、または５であり、
または式中、（ｉ）Ｒ^７およびＲ^８、または（ｉｉ）Ｒ^９およびＲ^１０は共に、任意で置換された５員または６員ヘテロシクロアルキルまたはヘテロアリールを形成し、ヘテロシクロアルキルまたはヘテロアリールは、Ｎ、ＯおよびＳから選択される１～３個のヘテロ原子を含み、
式中、Ｒ^１１、Ｒ^１２、Ｒ^１３、およびＲ^１４はそれぞれ独立してＨ、メチル、エチル、またはプロピルから選択され、
式中、Ｒ^Ａ、Ｒ^Ｂ、Ｒ^ＣおよびＲ^Ｄはそれぞれ独立して、任意で置換された（Ｃ_６－Ｃ_２０）アルキル、任意で置換された（Ｃ_６－Ｃ_２０）アルケニル、任意で置換された（Ｃ_６－Ｃ_２０）アルキニル、任意で置換された（Ｃ_６－Ｃ_２０）アシル、任意で置換された－ＯＣ（Ｏ）アルキル、任意で置換された－ＯＣ（Ｏ）アルケニル、任意で置換された（Ｃ_１－Ｃ_６）モノアルキルアミノ、任意で置換された（Ｃ_１－Ｃ_６）ジアルキルアミノ、任意で置換された（Ｃ_１－Ｃ_６）アルコキシ、－ＯＨ、－ＮＨ_２から選択され、
式中、Ｒ^７、Ｒ^８、Ｒ^９、Ｒ^１０のうちの少なくとも一つは、それぞれＲ^Ａ、Ｒ^Ｂ、Ｒ^ＣまたはＲ^Ｄ部分を含み、Ｒ^Ａ、Ｒ^Ｂ、Ｒ^ＣまたはＲ^Ｄは独立して、任意で置換された（Ｃ_６－Ｃ_２０）アルキル、任意で置換された（Ｃ_６－Ｃ_２０）アルケニル、任意で置換された（Ｃ_６－Ｃ_２０）アルキニル、任意で置換された（Ｃ_６－Ｃ_２０）アシル、任意で置換された－ＯＣ（Ｏ）（Ｃ_６－Ｃ_２０）アルキルまたは任意で置換された－ＯＣ（Ｏ）（Ｃ_６－Ｃ_２０）アルケニルから選択される化合物、
またはその薬学的に許容可能な塩を含む。 Compounds of the Invention Provided herein are compounds that are cationic lipids. For example, a cationic lipid of the invention is a compound having a structure according to formula (I):

wherein L ₁ is a bond, (C ₁ -C ₆ )alkyl or (C ₂ -C ₆ )alkenyl;
wherein X is O or S;
wherein R ¹ , R ² , R ³ , R ⁴ and R ⁵ are each independently H, OH, optionally substituted (C ₁ -C ₆ )alkyl, optionally substituted (C ₂ - C ₆ )alkenyl, optionally substituted (C ₂ -C ₆ )alkynyl, optionally substituted (C ₁ -C ₆ )alkoxy and —OC(O)R′;
wherein at least one of R ¹ , R ² , R ³ , R ⁴ or R ⁵ is —OC(O)R′;
wherein R' is

wherein ^R6 is

wherein m and p are each independently 0, 1, 2, 3, 4, or 5;
wherein R ⁷ is H, optionally substituted (C ₁ -C ₆ )alkyl, optionally substituted (C ₂ -C ₆ )alkenyl, optionally substituted (C ₂ -C ₆ )alkynyl , optionally substituted (C ₁ -C ₆ )acyl, —(CH ₂ ) _k R ^A or —(CH ₂ ) _k CH(OR ¹¹ )R ^A ;
wherein R ⁸ is H, optionally substituted (C ₁ -C ₆ )alkyl, optionally substituted (C ₂ -C ₆ )alkenyl, optionally substituted (C ₂ -C ₆ )alkynyl , optionally substituted (C ₁ -C ₆ )acyl, —(CH ₂ ) _n R ^B or —(CH ₂ ) _n CH(OR ¹² )R ^B ;
wherein R ⁹ is H, optionally substituted (C ₁ -C ₆ )alkyl, optionally substituted (C ₂ -C ₆ )alkenyl, optionally substituted (C ₂ -C ₆ )alkynyl , optionally substituted (C ₁ -C ₆ )acyl, —(CH ₂ ) _q R ^C or —(CH ₂ ) _q CH(OR ¹³ )R ^C ;
wherein R ¹⁰ is H, optionally substituted (C ₁ -C ₆ )alkyl, optionally substituted (C ₂ -C ₆ )alkenyl, optionally substituted (C ₂ -C ₆ )alkynyl , optionally substituted (C ₁ -C ₆ )acyl, —(CH ₂ ) _r R ^D or —(CH ₂ ) _r CH(OR ¹⁴ )R ^D ;
wherein k, n, q and r are each independently 1, 2, 3, 4, or 5;
or wherein (i) R ⁷ and R ⁸ or (ii) R ⁹ and R ¹⁰ together form an optionally substituted 5- or 6-membered heterocycloalkyl or heteroaryl, heterocycloalkyl or heterocycloalkyl aryl contains 1 to 3 heteroatoms selected from N, O and S;
wherein R ¹¹ , R ¹² , R ¹³ and R ¹⁴ are each independently selected from H, methyl, ethyl or propyl;
wherein R ^A , R ^B , R ^C and R ^D are each independently optionally substituted (C ₆ -C ₂₀ )alkyl, optionally substituted (C ₆ -C ₂₀ )alkenyl, optionally substituted (C ₆ -C ₂₀ )alkynyl, optionally substituted (C ₆ -C ₂₀ )acyl, optionally substituted —OC(O)alkyl, optionally substituted —OC(O)alkenyl, optionally substituted (C ₁ -C ₆ )monoalkylamino, optionally substituted (C ₁ -C ₆ )dialkylamino, optionally substituted (C ₁ -C ₆ )alkoxy, —OH, —NH selected from ₂ ,
wherein at least one of R ⁷ , R ⁸ , R ⁹ , R ¹⁰ each comprises a R ^A , R ^B , R ^C or R ^D moiety and R ^A , R ^B , R ^C or R ^D is Independently optionally substituted (C ₆ -C ₂₀ )alkyl, optionally substituted (C ₆ -C ₂₀ )alkenyl, optionally substituted (C ₆ -C ₂₀ )alkynyl, optionally substituted (C ₆ -C ₂₀ )acyl, optionally substituted —OC(O)(C ₆ -C ₂₀ )alkyl or optionally substituted —OC(O)(C ₆ -C ₂₀ )alkenyl compounds that
or a pharmaceutically acceptable salt thereof.

In embodiments, any alkyl, alkenyl, alkynyl, acyl, alkoxy, monoalkylamino, dialkylamino, heterocycloalkyl or heteroaryl is (C1-C6)alkyl, (C2-C6)alkenyl, (C2- C6) alkynyl, (C1-C6) acyl, (C1-C6) alkoxy, halogen, -COR, -COH, -COR, -CN, -OH, -OR, -OCOR, -OCO2R, -NH, -NHR, optionally substituted with one or more substituents selected from the group consisting of -N(R)2, -SR or -SO2R, or two geminal hydrogens on the carbon atom are replaced with the group =NH, Each instance of R is independently C1-C10 aliphatic alkyl.

In embodiments, L1 is a bond.

In several embodiments, L1 is (C1-C6)alkyl.

In several embodiments, L1 is (C2-C6)alkenyl.

In several embodiments, L1 is C2 alkenyl.

In embodiments, RA and RB are the same. In some embodiments, RC and RD are the same. In some embodiments, RA and RB are the same and RC and RD are the same.

In embodiments, RA and RB are different. In some embodiments, RC and RD are different. In embodiments, RA and RB are different and RC and RD are different.

In some embodiments, RA, RB, RC and RD are the same.

In some embodiments, RA, RB, RC and RD are different.

In embodiments, each RA, RB, RC or RD is independently optionally substituted (C6-C20) alkyl, optionally substituted (C6-C20) alkenyl, optionally substituted ( C6-C20)alkynyl, optionally substituted (C6-C20)acyl, optionally substituted -OC(O)(C6-C20)alkyl or optionally substituted -OC(O)(C6-C20) selected from alkenyl;

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

In embodiments, RA and RB are each independently optionally substituted (C6-C20)alkyl, optionally substituted (C6-C20)alkenyl, optionally substituted (C6-C20)alkynyl is selected from

In embodiments, RA and RB are the same and are from optionally substituted (C6-C20)alkyl, optionally substituted (C6-C20)alkenyl, optionally substituted (C6-C20)alkynyl selected.

In embodiments, each RA and RB is independently optionally substituted (C6-C20)alkyl.

In embodiments, RA and RB are the same and are optionally substituted (C6-C20)alkyl.

In embodiments, each RA and RB is independently an optionally substituted (C6-C20)alkenyl.

In embodiments, RA and RB are the same and are optionally substituted (C6-C20)alkenyl.

In embodiments, each RA and RB is independently an optionally substituted (C6-C20)alkynyl.

In embodiments, RA and RB are the same and are optionally substituted (C6-C20)alkynyl.

In embodiments, each RA and RB is independently optionally substituted (C6-C20) acyl.

In embodiments, RA and RB are the same, optionally substituted (C6-C20) acyl.

In embodiments, each RA and RB is independently an optionally substituted -OC(O)(C6-C20)alkyl.

In embodiments, RA and RB are the same and are optionally substituted -OC(O)(C6-C20)alkyl.

In embodiments, each RA and RB is independently an optionally substituted -OC(O)(C6-C20)alkenyl.

In embodiments, RA and RB are the same and are optionally substituted -OC(O)(C6-C20)alkenyl.

In embodiments, R7=-(CH2)kCH(OR11)RA, R8 is -(CH2)nCH(OR12)RB, and RA and RB are each independently selected from:

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

In embodiments, R7=-(CH2)kCH(OR11)RA, R8 is -(CH2)nCH(OR12)RB, and both RA and RB are C8H17.

In embodiments, R7=-(CH2)kCH(OR11)RA, R8 is -(CH2)nCH(OR12)RB, and both RA and RB are C10H21.

In embodiments, R7=-(CH2)kCH(OR11)RA, R8 is -(CH2)nCH(OR12)RB, and both RA and RB are C12H25.

である。 In embodiments, R7=-(CH2)kCH(OR11)RA, R8 is -(CH2)nCH(OR12)RB, and RA and RB are both

is.

In some embodiments, X is O.

In some embodiments, X is S.

In embodiments, only one of R1, R2, R3, R4, and R5 is -OC(O)R'. In embodiments, only one of R1, R2, R3, R4, and R5 is -OC(O)R' and none of R1, R2, R3, R4, or R5 is OH.

In embodiments, two of R1, R2, R3, R4, and R5 are -OC(O)R'. In embodiments, two of R1, R2, R3, R4, and R5 are -OC(O)R' and none of R1, R2, R3, R4, or R5 is OH.

In embodiments, three of R1, R2, R3, R4, and R5 are -OC(O)R'.

In some embodiments, R1 is -OC(O)R'. In several embodiments, R5 is -OC(O)R'. In several embodiments, both R1 and R5 are -OC(O)R'.

In some embodiments, R2 is -OC(O)R'. In several embodiments, R4 is -OC(O)R'. In several embodiments, both R2 and R4 are -OC(O)R'.

In some embodiments, R3 is -OC(O)R'.

In embodiments, R3 is -OC(O)R' and R2 is OMe.

In embodiments, L1 is a bond, R3 is -OC(O)R' and R2 is OMe.

In embodiments, R3 is -OC(O)R' and R2 and R4 are OMe.

In several embodiments, L1 is a bond, R3 is -OC(O)R', and R2 and R4 are OMe.

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

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

In embodiments, R7 is -(CH2)kCH(OR11)RA.

In several embodiments, R7 is -(CH2)1CH(OR11)RA.

In several embodiments, R7 is -(CH2)1CH(OH)RA.

In several embodiments, R8 is -(CH2)nCH(OR12)RB.

In several embodiments, R8 is -(CH2)1CH(OR12)RB.

In several embodiments, R8 is -(CH2)1CH(OH)RB.

In embodiments, R7 is -(CH2)kCH(OR11)RA and R8 is -(CH2)nCH(OR12)RB.

In embodiments, R7 is -(CH2)1CH(OR11)RA and R8 is -(CH2)1CH(OR12)RB.

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

In embodiments, R7 and R8 are each optionally substituted (C1-C6)alkyl, for example ( _C1 - _C6 )alkyl substituted with —CO ₂ R ^aa , wherein R ^aa is C ₁ -C ₅₀ alkyl. In embodiments, each of R ⁷ and R ⁸ is (C ₁ -C ₆ )alkyl substituted with —CO ₂ R ^aa , wherein R ^aa is C ₁ -C ₄₀ alkyl. In embodiments, R ⁷ and R ⁸ are each (C ₁ -C ₆ )alkyl substituted with —CO ₂ R ^aa , wherein R ^aa is C ₁ -C ₃₀ alkyl. In embodiments, each of R ⁷ and R ⁸ is (C ₁ -C ₆ )alkyl substituted with —CO ₂ R ^aa , wherein R ^aa is C ₁ -C ₂₀ alkyl.

In embodiments, R7 and R8 are the same and each is optionally substituted (C1-C6)alkyl, for example (C1-C6)alkyl substituted with —CO2Raa, wherein Raa is C1-C50 alkyl. In embodiments, R7 and R8 are the same and each is (C1-C6)alkyl substituted with -CO2Raa, wherein Raa is C1-C40alkyl. In embodiments, R7 and R8 are the same and each is (C1-C6)alkyl substituted with -CO2Raa, wherein Raa is C1-C30 alkyl. In embodiments, each R7 and R8 is (C1-C6)alkyl substituted with -CO2Raa, wherein Raa is C1-C20alkyl.

である。 In some embodiments, each of R7 and R8 is

is.

である。 In some embodiments, each of R7 and R8 is

is.

In several embodiments, R9 and R10 are each independently H, optionally substituted (C1-C6)alkyl, optionally substituted (C2-C6)alkenyl, optionally substituted (C2-C6 ) alkynyl.

In embodiments, each R9 and R10 is independently optionally substituted (C1-C6)alkyl or optionally substituted (C2-C6)alkenyl.

In embodiments, both R9 and R10 are optionally substituted (C1-C6)alkyl or optionally substituted (C2-C6)alkenyl.

In embodiments, both R9 and R10 are optionally substituted (C1-C6)alkyl.

In embodiments, both R9 and R10 are -CH3.

In embodiments, R7 is -(CH2)kCH(OR11)RA, R8 is -(CH2)nCH(OR12)RB, and both R9 and R10 are -CH3.

In embodiments, R7 is -(CH2)1CH(OR11)RA, R8 is -(CH2)1CH(OR12)RB, and R9 and R10 are both -CH3.

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

In embodiments, R7 is -(CH2)kCH(OR11)RA, R8 is -(CH2)nCH(OR12)RB, RA and RB are both C8H17, R9 and R10 are both -CH3 is.

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

In embodiments, R7 is -(CH2)kCH(OR11)RA, R8 is -(CH2)nCH(OR12)RB, RA and RB are both C10H21, R9 and R10 are both -CH3 is.

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

In embodiments, R7 is -(CH2)kCH(OR11)RA, R8 is -(CH2)nCH(OR12)RB, RA and RB are both C12H25, R9 and R10 are both -CH3 is.

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

In embodiments, R7 is -(CH2)kCH(OR11)RA, R8 is -(CH2)nCH(OR12)RB, RA and RB are both C16H29, and R9 and R10 are both -CH3 is.

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

In some embodiments, p, q and r are the same. In embodiments, one or more of p, q, and r are different. In embodiments, q and r are the same and p is different. In embodiments, p and q are the same and r is different. In embodiments, p and r are the same and q is different. In embodiments, p, q and r are different.

In some embodiments, k, m and n are the same. In embodiments, one or more of k, m and n are different. In some embodiments, k and m are the same and n is different. In some embodiments, m and n are the same and k is different. In some embodiments, k and n are the same and m is different. In embodiments, k, m and n are different.

In embodiments, m is 1, 2, 3, 4 or 5. In some embodiments, m is 0. In some embodiments, m is one. In embodiments, m is two. In embodiments, m is three. In embodiments, m is four. In embodiments, m is five. In embodiments, m is 0, 1, 2, 3, or 4.

In embodiments, p is 1, 2, 3, 4 or 5. In some embodiments, p is 0. In embodiments, p is one. In embodiments, p is two. In embodiments, p is three. In embodiments, p is four. In embodiments, p is five. In embodiments, p is 0, 1, 2, 3, or 4.

In embodiments, m is two and p is two.

In embodiments, m is three and p is two.

In embodiments, k and n=1 and m=2.

In embodiments, k and n=1 and m=3.

In embodiments, q and r=1 and p=2.

In embodiments, k, n, q, and r are each =1, m=2 or 3, and p=2.

In some embodiments, R' is

ｋおよびｎ＝１、ｍ＝２または３である。 In some embodiments, R' is

k and n=1, m=2 or 3;

ｋおよびｎ＝１、ｍ＝２である。 In some embodiments, R' is

k and n=1, m=2.

ｋおよびｎ＝１、ｍ＝３である。 In some embodiments, R' is

k and n=1, m=3.

であり、Ｒ^１１およびＲ^１２はＨである。 In some embodiments, R' is

and R ¹¹ and R ¹² are H.

であり、ｋおよびｎ＝１、ｍ＝２または３、Ｒ^１１およびＲ^１２はＨである。 In some embodiments, R' is

and k and n=1, m=2 or 3, R ¹¹ and R ¹² are H.

であり、ｋおよびｎ＝１、ｍ＝２、Ｒ^１１およびＲ^１２はＨである。 In some embodiments, R' is

and k and n=1, m=2, R ¹¹ and R ¹² are H.

であり、ｋおよびｎ＝１、ｍ＝３、Ｒ^１１およびＲ^１２はＨである。 In some embodiments, R' is

and k and n=1, m=3, R ¹¹ and R ¹² are H.

In some embodiments, R' is

Ｒ^ＡおよびＲ^Ｂは、段落［０１０４］～［０１２９］のいずれかに定義されるとおりであってもよい。 In any of the above embodiments, when R' has the structure

R ^A and R ^B may be as defined in any of paragraphs [0104]-[0129].

In some embodiments, ^R6 is

であり、ｑおよびｒ＝１であり、ｐ＝２である。 In several embodiments, ^R6 is

, q and r=1 and p=2.

であり、Ｒ^１３およびＲ^１４はＨである。 In several embodiments, ^R6 is

and R ¹³ and R ¹⁴ are H.

であり、ｑおよびｒ＝１であり、ｐ＝２であり、Ｒ^１３およびＲ^１４はＨである。 In several embodiments, ^R6 is

, q and r=1, p=2 and R ¹³ and R ¹⁴ are H.

In embodiments, ^R6 is selected from the group consisting of:

In embodiments, ^R6 is selected from the group consisting of:

In embodiments, ^R6 is selected from the group consisting of:

In some embodiments, ^R6 is

In some embodiments, ^R6 is

In some embodiments, ^R6 is

In embodiments, ^R6 is selected from the group consisting of:

In some embodiments, ^R6 is

In some embodiments, ^R6 is

In some embodiments, ^R6 is

In some embodiments, ^R6 is

In some embodiments, ^R6 is

In several embodiments, R ⁶ and R ^' are the same.

であり、Ｒ^’は

であり、ｍは２であり、ｐは２である。 In several embodiments, ^R6 is

and R ^' is

, m is 2 and p is 2.

であり、Ｒ^’は

であり、ｍは３であり、ｐは２である。 In several embodiments, ^R6 is

and R ^' is

, m is 3 and p is 2.

であり、Ｒ^’は

である。 In embodiments, L ₁ is a bond, R ³ is -OC(O)R', R ² and R ⁴ are OMe, and R ⁶ is

and R ^′ is

is.

である。 In embodiments, L ₁ is a bond, R ³ is -OC(O)R', R ² and R ⁴ are OMe, and R ⁶ is

is.

であり、Ｒ^’は

である。 In embodiments, X=O, L ₁ is a bond, R ³ is -OC(O)R', R ² and R ⁴ are OMe, R ⁶ is

and R ^' is

is.

であり、Ｒ^’は

である。 In embodiments, X=O, L ₁ is a bond, R ³ is -OC(O)R', R ² and R ⁴ are OMe, R ¹ and R ⁵ are H Yes, ^R6 is

and R ^′ is

is.

であり、Ｒ^’は

である。 In embodiments, X=O, L ₁ is a bond, R ³ is -OC(O)R', R ² and R ⁴ are OMe, R ⁶ is

and R ^' is

is.

であり、Ｒ^’は

である。 In embodiments, X=O, L ₁ is a bond, R ³ is -OC(O)R', R ² and R ⁴ are OMe, R ¹ and R ⁵ are H Yes, ^R6 is

and R ^' is

is.

であり、Ｒ^’は

である。 In embodiments, L ₁ is a bond, R ³ is -OC(O)R', R ² is OMe, and R ⁶ is

and R ^′ is

is.

であり、Ｒ^’は

である。 In embodiments, X=O, L ₁ is a bond, R ³ is -OC(O)R', R ² is OMe, R ⁶ is

and R ^′ is

is.

であり、Ｒ^’は

である。 In embodiments, X=O, L ₁ is a bond, R ³ is -OC(O)R', R ² is OMe, R ¹ , R ⁴ and R ⁵ are H and R ⁶ is

and R ^' is

is.

であり、Ｒ^’は

である。 In embodiments, L ₁ is a bond, R ³ is -OC(O)R', R ² is OMe, and R ⁶ is

and R ^′ is

is.

であり、Ｒ^’は

である。 In embodiments, X=O, L ₁ is a bond, R ³ is -OC(O)R', R ² is OMe, R ⁶ is

and R ^′ is

is.

であり、Ｒ^’は

である。 In embodiments, X=O, L ₁ is a bond, R ³ is -OC(O)R', R ² is OMe, R ¹ , R ⁴ and R ⁵ are H and R ⁶ is

and R ^' is

is.

であり、Ｒ^’は

である。 In embodiments, L ₁ is C ₂ alkenyl, R ³ is -OC(O)R', R ² and R ⁴ are OMe, and R ⁶ is

and R ^′ is

is.

であり、Ｒ^’は

である。 In embodiments, L ₁ is C ₂ alkenyl, R ³ is -OC(O)R', R ² and R ⁴ are OMe, and R ⁶ is

and R ^' is

is.

であり、Ｒ^’は

である。 In embodiments, X═O, L ₁ is C ₂ alkenyl, R ³ is —OC(O)R′, R ² and R ⁴ are OMe, and R ⁶ is

and R ^′ is

is.

であり、Ｒ^’は

である。 In embodiments, X═O, L ₁ is C ₂ alkenyl, R ³ is —OC(O)R′, R ² and R ⁴ are OMe, R ¹ and R ⁵ are H and ^R6 is

and R ^′ is

is.

であり、Ｒ^’は

である。 In embodiments, X═O, L ₁ is C ₂ alkenyl, R ³ is —OC(O)R′, R ² and R ⁴ are OMe, and R ⁶ is

and R ^′ is

is.

であり、Ｒ^’は

である。 In embodiments, X=O, L ₁ is C ₂ alkenyl, R ³ is -OC(O)R', R ² and R ⁴ are OMe, R ¹ and R ⁵ are H and ^R6 is

and R ^′ is

is.

であり、Ｒ^’は

であり、Ｒ^７およびＲ^８はそれぞれ、－ＣＯ_２Ｒ^ａａで置換された（Ｃ_１－Ｃ_６）アルキルであり、式中、Ｒ^ａａはＣ_１－Ｃ_５０アルキルである。 In several embodiments, ^R6 is

and R ^' is

and R ⁷ and R ⁸ are each (C ₁ -C ₆ )alkyl substituted with —CO ₂ R ^aa , where R ^aa is C ₁ -C ₅₀ alkyl.

であり、Ｒ^’は

であり、Ｒ^７およびＲ^８はそれぞれ、－ＣＯ_２Ｒ^ａａで置換された（Ｃ_１－Ｃ_６）アルキルであり、式中、Ｒ^ａａは、Ｃ_１－Ｃ_５０アルキルであり、ｍは２であり、ｐは２である。複数の実施形態では、Ｒ^７およびＲ^８は同じである。一部の実施形態では、Ｌ_１は結合であり、Ｒ^３は－ＯＣ（Ｏ）Ｒ’であり、Ｒ^２およびＲ^４はＯＭｅである。 In several embodiments, ^R6 is

and R ^′ is

and R ⁷ and R ⁸ are each (C ₁ -C ₆ )alkyl substituted with —CO ₂ R ^aa wherein R ^aa is C ₁ -C ₅₀ alkyl and m is 2 and p is 2. In several embodiments, ^R7 and ^R8 are the same. In some embodiments, L ₁ is a bond, R ³ is -OC(O)R', and R ² and R ⁴ are OMe.

であり、Ｒ^’は

であり、Ｒ^７およびＲ^８はそれぞれ、－ＣＯ_２Ｒ^ａａで置換された（Ｃ_１－Ｃ_６）アルキルであり、式中、Ｒ^ａａはＣ_１－Ｃ_５０アルキルである。複数の実施形態では、Ｒ^７およびＲ^８は同じである。 In embodiments, L ₁ is a bond, R ³ is -OC(O)R', R ² and R ⁴ are OMe, and R ⁶ is

and R ^' is

and R ⁷ and R ⁸ are each (C ₁ -C ₆ )alkyl substituted with —CO ₂ R ^aa , where R ^aa is C ₁ -C ₅₀ alkyl. In several embodiments, ^R7 and ^R8 are the same.

であり、Ｒ^’は

であり、Ｒ^７およびＲ^８はそれぞれ、－ＣＯ_２Ｒ^ａａで置換された（Ｃ_１－Ｃ_６）アルキルであり、式中、Ｒ^ａａはＣ_１－Ｃ_５０アルキルであり、ｍは２である。複数の実施形態では、Ｒ^７およびＲ^８は同じである。 In embodiments, L ₁ is a bond, R ³ is -OC(O)R', R ² and R ⁴ are OMe, and R ⁶ is

and R ^′ is

and R ⁷ and R ⁸ are each (C ₁ -C ₆ )alkyl substituted with —CO ₂ R ^aa wherein R ^aa is C ₁ -C ₅₀ alkyl and m is 2 be. In several embodiments, ^R7 and ^R8 are the same.

であり、Ｒ^’は

であり、Ｒ^７およびＲ^８はそれぞれ、－ＣＯ_２Ｒ^ａａで置換された（Ｃ_１－Ｃ_６）アルキルであり、式中、Ｒ^ａａはＣ_１－Ｃ_５０アルキルである。複数の実施形態では、Ｒ^７およびＲ^８は同じである。一部の実施形態では、ｍは２である。 In embodiments, X=O, L ₁ is a bond, R ³ is -OC(O)R', R ² and R ⁴ are OMe, R ⁶ is

and R ^' is

and R ⁷ and R ⁸ are each (C ₁ -C ₆ )alkyl substituted with —CO ₂ R ^aa , where R ^aa is C ₁ -C ₅₀ alkyl. In several embodiments, ^R7 and ^R8 are the same. In some embodiments, m is two.

であり、Ｒ^’は

であり、Ｒ^７およびＲ^８はそれぞれ、－ＣＯ_２Ｒ^ａａで置換された（Ｃ_１－Ｃ_６）アルキルであり、式中、Ｒ^ａａはＣ_１－Ｃ_５０アルキルである。複数の実施形態では、Ｒ^７およびＲ^８は同じである。一部の実施形態では、ｍは２である。 In embodiments, X=O, L ₁ is a bond, R ³ is -OC(O)R', R ² and R ⁴ are OMe, R ¹ and R ⁵ are H Yes, ^R6 is

and R ^' is

and R ⁷ and R ⁸ are each (C ₁ -C ₆ )alkyl substituted with —CO ₂ R ^aa , where R ^aa is C ₁ -C ₅₀ alkyl. In several embodiments, ^R7 and ^R8 are the same. In some embodiments, m is two.

であり、Ｒ^’は

であり、Ｒ^７およびＲ^８はそれぞれ、－ＣＯ_２Ｒ^ａａで置換された（Ｃ_１－Ｃ_６）アルキルであり、式中、Ｒ^ａａはＣ_１－Ｃ_５０アルキルであり、ｍは２である。複数の実施形態では、Ｒ^７およびＲ^８は同じである。 In embodiments, X=O, L ₁ is a bond, R ³ is -OC(O)R', R ² and R ⁴ are OMe, R ⁶ is

and R ^' is

and R ⁷ and R ⁸ are each (C ₁ -C ₆ )alkyl substituted with —CO ₂ R ^aa wherein R ^aa is C ₁ -C ₅₀ alkyl and m is 2 be. In several embodiments, ^R7 and ^R8 are the same.

であり、Ｒ^’は

であり、Ｒ^７およびＲ^８はそれぞれ、－ＣＯ_２Ｒ^ａａで置換された（Ｃ_１－Ｃ_６）アルキルであり、式中、Ｒ^ａａはＣ_１－Ｃ_５０アルキルであり、ｍは２である。複数の実施形態では、Ｒ^７およびＲ^８は同じである。 In embodiments, X=O, L ₁ is a bond, R ³ is -OC(O)R', R ² and R ⁴ are OMe, R ¹ and R ⁵ are H Yes, ^R6 is

and R ^' is

and R ⁷ and R ⁸ are each (C ₁ -C ₆ )alkyl substituted with —CO ₂ R ^aa wherein R ^aa is C ₁ -C ₅₀ alkyl and m is 2 be. In several embodiments, ^R7 and ^R8 are the same.

であり、Ｒ^’は

であり、Ｒ^７およびＲ^８はそれぞれ、－ＣＯ_２Ｒ^ａａで置換された（Ｃ_１－Ｃ_６）アルキルであり、式中、Ｒ^ａａはＣ_１－Ｃ_５０アルキルである。複数の実施形態では、Ｒ^７およびＲ^８は同じである。一部の実施形態では、ｍは２である。 In embodiments, L ₁ is a bond, R ³ is -OC(O)R', R ² is OMe, and R ⁶ is

and R ^' is

and R ⁷ and R ⁸ are each (C ₁ -C ₆ )alkyl substituted with —CO ₂ R ^aa , where R ^aa is C ₁ -C ₅₀ alkyl. In several embodiments, ^R7 and ^R8 are the same. In some embodiments, m is two.

であり、Ｒ^’は

であり、Ｒ^７およびＲ^８はそれぞれ、－ＣＯ_２Ｒ^ａａで置換された（Ｃ_１－Ｃ_６）アルキルであり、式中、Ｒ^ａａはＣ_１－Ｃ_５０アルキルである。複数の実施形態では、Ｒ^７およびＲ^８は同じである。一部の実施形態では、ｍは２である。 In embodiments, X=O, L ₁ is a bond, R ³ is -OC(O)R', R ² is OMe, R ⁶ is

and R ^' is

and R ⁷ and R ⁸ are each (C ₁ -C ₆ )alkyl substituted with —CO ₂ R ^aa , where R ^aa is C ₁ -C ₅₀ alkyl. In several embodiments, ^R7 and ^R8 are the same. In some embodiments, m is two.

であり、Ｒ^７およびＲ^８はそれぞれ、－ＣＯ_２Ｒ^ａａで置換された（Ｃ_１－Ｃ_６）アルキルであり、式中、Ｒ^ａａはＣ_１－Ｃ_５０アルキルである。複数の実施形態では、Ｒ^７およびＲ^８は同じである。一部の実施形態では、ｍは２である。 In embodiments, X=O, L ₁ is a bond, R ³ is -OC(O)R', R ² is OMe, R ¹ , R ⁴ and R ⁵ are H and R ⁶ is

and R ⁷ and R ⁸ are each (C ₁ -C ₆ )alkyl substituted with —CO ₂ R ^aa , where R ^aa is C ₁ -C ₅₀ alkyl. In several embodiments, ^R7 and ^R8 are the same. In some embodiments, m is two.

であり、Ｒ^７およびＲ^８はそれぞれ、－ＣＯ_２Ｒ^ａａで置換された（Ｃ_１－Ｃ_６）アルキルであり、式中、Ｒ^ａａはＣ_１－Ｃ_５０アルキルであり、ｍは２である。複数の実施形態では、Ｒ^７およびＲ^８は同じである。 In embodiments, L ₁ is a bond, R ³ is -OC(O)R', R ² is OMe, and R ⁶ is

and R ⁷ and R ⁸ are each (C ₁ -C ₆ )alkyl substituted with —CO ₂ R ^aa wherein R ^aa is C ₁ -C ₅₀ alkyl and m is 2 be. In several embodiments, ^R7 and ^R8 are the same.

であり、Ｒ^７およびＲ^８はそれぞれ、－ＣＯ_２Ｒ^ａａで置換された（Ｃ_１－Ｃ_６）アルキルであり、式中、Ｒ^ａａはＣ_１－Ｃ_５０アルキルであり、ｍは２である。複数の実施形態では、Ｒ^７およびＲ^８は同じである。 In embodiments, X=O, L ₁ is a bond, R ³ is -OC(O)R', R ² is OMe, R ¹ , R ⁴ and R ⁵ are H Yes, ^R6 is

and R ⁷ and R ⁸ are each (C ₁ -C ₆ )alkyl substituted with —CO ₂ R ^aa wherein R ^aa is C ₁ -C ₅₀ alkyl and m is 2 be. In several embodiments, ^R7 and ^R8 are the same.

であり、Ｒ^’は

であり、Ｒ^７およびＲ^８はそれぞれ、－ＣＯ_２Ｒ^ａａで置換された（Ｃ_１－Ｃ_６）アルキルであり、式中、Ｒ^ａａはＣ_１－Ｃ_５０アルキルである。複数の実施形態では、Ｒ^７およびＲ^８は同じである。 In embodiments, L ₁ is C ₂ alkenyl, R ³ is -OC(O)R', R ² and R ⁴ are OMe, and R ⁶ is

and R ^' is

and R ⁷ and R ⁸ are each (C ₁ -C ₆ )alkyl substituted with —CO ₂ R ^aa , where R ^aa is C ₁ -C ₅₀ alkyl. In several embodiments, ^R7 and ^R8 are the same.

であり、Ｒ^’は

であり、Ｒ^７およびＲ^８はそれぞれ、－ＣＯ_２Ｒ^ａａで置換された（Ｃ_１－Ｃ_６）アルキルであり、式中、Ｒ^ａａはＣ_１－Ｃ_５０アルキルであり、ｍは２である。複数の実施形態では、Ｒ^７およびＲ^８は同じである。 In embodiments, L ₁ is C ₂ alkenyl, R ³ is -OC(O)R', R ² and R ⁴ are OMe, and R ⁶ is

and R ^' is

and R ⁷ and R ⁸ are each (C ₁ -C ₆ )alkyl substituted with —CO ₂ R ^aa wherein R ^aa is C ₁ -C ₅₀ alkyl and m is 2 be. In several embodiments, ^R7 and ^R8 are the same.

であり、Ｒ^’は

であり、Ｒ^７およびＲ^８はそれぞれ、－ＣＯ_２Ｒ^ａａで置換された（Ｃ_１－Ｃ_６）アルキルであり、式中、Ｒ^ａａはＣ_１－Ｃ_５０アルキルである。複数の実施形態では、Ｒ^７およびＲ^８は同じである。 In embodiments, X═O, L ₁ is C ₂ alkenyl, R ³ is —OC(O)R′, R ² and R ⁴ are OMe, and R ⁶ is

and R ^′ is

and R ⁷ and R ⁸ are each (C ₁ -C ₆ )alkyl substituted with —CO ₂ R ^aa , where R ^aa is C ₁ -C ₅₀ alkyl. In several embodiments, ^R7 and ^R8 are the same.

であり、Ｒ^’は

であり、Ｒ^７およびＲ^８はそれぞれ、－ＣＯ_２Ｒ^ａａで置換された（Ｃ_１－Ｃ_６）アルキルであり、式中、Ｒ^ａａはＣ_１－Ｃ_５０アルキルである。複数の実施形態では、Ｒ^７およびＲ^８は同じである。 In embodiments, X=O, L ₁ is C ₂ alkenyl, R ³ is -OC(O)R', R ² and R ⁴ are OMe, R ¹ and R ⁵ are H and ^R6 is

and R ^' is

and R ⁷ and R ⁸ are each (C ₁ -C ₆ )alkyl substituted with —CO ₂ R ^aa , where R ^aa is C ₁ -C ₅₀ alkyl. In several embodiments, ^R7 and ^R8 are the same.

であり、Ｒ^’は

であり、Ｒ^７およびＲ^８はそれぞれ、－ＣＯ_２Ｒ^ａａで置換された（Ｃ_１－Ｃ_６）アルキルであり、式中、Ｒ^ａａはＣ_１－Ｃ_５０アルキルであり、ｍは２である。複数の実施形態では、Ｒ^７およびＲ^８は同じである。 In embodiments, X═O, L ₁ is C ₂ alkenyl, R ³ is —OC(O)R′, R ² and R ⁴ are OMe, and R ⁶ is

and R ^′ is

and R ⁷ and R ⁸ are each (C ₁ -C ₆ )alkyl substituted with —CO ₂ R ^aa wherein R ^aa is C ₁ -C ₅₀ alkyl and m is 2 be. In several embodiments, ^R7 and ^R8 are the same.

であり、Ｒ^’は

であり、Ｒ^７およびＲ^８はそれぞれ、－ＣＯ_２Ｒ^ａａで置換された（Ｃ_１－Ｃ_６）アルキルであり、式中、Ｒ^ａａはＣ_１－Ｃ_５０アルキルであり、ｍは２である。複数の実施形態では、Ｒ^７およびＲ^８は同じである。 In embodiments, X=O, L ₁ is C ₂ alkenyl, R ³ is -OC(O)R', R ² and R ⁴ are OMe, R ¹ and R ⁵ are H and ^R6 is

and R ^′ is

and R ⁷ and R ⁸ are each (C ₁ -C ₆ )alkyl substituted with —CO ₂ R ^aa wherein R ^aa is C ₁ -C ₅₀ alkyl and m is 2 be. In several embodiments, ^R7 and ^R8 are the same.

In any of the above embodiments, R ⁷ and R ⁸ are (C ₁ -C ₆ )alkyl substituted with —CO ₂ R ^aa , wherein R ^aa is C ₁ -C ₅₀ alkyl. , R ^aa may alternatively be C ₁ -C ₄₀ alkyl.

In any of the above embodiments, R ⁷ and R ⁸ are (C ₁ -C ₆ )alkyl substituted with —CO ₂ R ^aa , wherein R ^aa is C ₁ -C ₅₀ alkyl , R ^aa may alternatively be C ₁ -C ₃₀ alkyl.

In any of the above embodiments, R ⁷ and R ⁸ are (C ₁ -C ₆ )alkyl substituted with —CO ₂ R ^aa , wherein R ^aa is C ₁ -C ₅₀ alkyl , R ^aa may alternatively be C ₁ -C ₂₀ alkyl.

であってもよい。 In any of the above embodiments, R ⁷ and R ⁸ are (C ₁ -C ₆ )alkyl substituted with —CO ₂ R ^aa , wherein R ^aa is C ₁ -C ₅₀ alkyl. , ^R7 and ^R8 are respectively

may be

であってもよい。 In any of the above embodiments, R ⁷ and R ⁸ are each (C ₁ -C ₆ )alkyl substituted with —CO ₂ R ^aa , wherein R ^aa is C ₁ -C ₅₀ alkyl. Yes, and R ⁷ and R ⁸ are each

may be

またはその薬学的に許容可能な塩を含み、式中Ｒ^１～Ｒ^６およびＸは、本明細書に既に定義されるとおりである。 In embodiments, the cationic lipids of the present invention are compounds having a structure according to formula (II):

or a pharmaceutically acceptable salt thereof, wherein R ¹ -R ⁶ and X are as previously defined herein.

またはその薬学的に許容可能な塩を含み、式中、Ｒ^’、Ｒ^６およびＸは、本明細書に既に定義されるとおりである。 In embodiments, the cationic lipid of the present invention is a compound having a structure according to Formula (IIA):

or a pharmaceutically acceptable salt thereof, wherein R ^' , ^R6 and X are as previously defined herein.

またはその薬学的に許容可能な塩を含む。 In embodiments, the cationic lipids of the present invention are compounds having structures according to formulas (IIB), (IIC), (IID), (IIE), (IIJ), or (IIK) below,

or a pharmaceutically acceptable salt thereof.

またはその薬学的に許容可能な塩を有する。 In embodiments, the cationic lipids of the present invention have the structure:

or a pharmaceutically acceptable salt thereof.

またはその薬学的に許容可能な塩を有する。 In embodiments, the cationic lipids of the present invention have the structure:

or a pharmaceutically acceptable salt thereof.

またはその薬学的に許容可能な塩を有する。 In embodiments, the cationic lipids of the present invention have the structure:

or a pharmaceutically acceptable salt thereof.

またはその薬学的に許容可能な塩を有する。 In embodiments, the cationic lipids of the present invention have the structure:

or a pharmaceutically acceptable salt thereof.

またはその薬学的に許容可能な塩を含み、式中、Ｒ^’、Ｒ^６およびＸは、本明細書に既に定義されるとおりである。 In embodiments, the cationic lipid of the present invention is a compound having a structure according to formula (IIF):

or a pharmaceutically acceptable salt thereof, wherein R ^' , ^R6 and X are as previously defined herein.

またはその薬学的に許容可能な塩を含み、式中、Ｒ^’、Ｒ^６およびＸは、本明細書に既に定義されるとおりである。 In embodiments, the cationic lipid of the present invention is a compound having a structure according to formula (IIG):

or a pharmaceutically acceptable salt thereof, wherein R ^' , ^R6 and X are as previously defined herein.

式中、ＹおよびＺのうちの一つはＯＨであり、他方は－ＯＣ（Ｏ）Ｒ’であり、またはＹおよびＺの両方がそれぞれ独立して、－ＯＣ（Ｏ）Ｒ’であり、式中、Ｒ^’、Ｒ^６およびＸは本明細書に既に定義されるとおりである。 In embodiments, the cationic lipids of the present invention comprise compounds having a structure according to formula (IIH):

wherein one of Y and Z is OH and the other is -OC(O)R', or both Y and Z are each independently -OC(O)R'; wherein R ^' , ^R6 and X are as previously defined herein.

またはその薬学的に許容可能な塩を含み、式中、Ｒ^１～Ｒ^６およびＸは、本明細書に既に定義されるとおりである。 In embodiments, the cationic lipids of the present invention are compounds having a structure according to formula (III):

or a pharmaceutically acceptable salt thereof, wherein R ¹ -R ⁶ and X are as previously defined herein.

またはその薬学的に許容可能な塩を含み、式中、Ｒ^’、Ｒ^６およびＸは、本明細書に既に定義されるとおりである。 In embodiments, the cationic lipid of the present invention is a compound having a structure according to formula (IIIA):

or a pharmaceutically acceptable salt thereof, wherein R ^' , ^R6 and X are as previously defined herein.

またはその薬学的に許容可能な塩を含み、式中、Ｒ^Ａ、Ｒ^Ｂおよびｐは、本明細書に既に定義されるとおりである。 In embodiments, the cationic lipid of the present invention is a compound having a structure according to formula (IIIB):

or a pharmaceutically acceptable salt thereof, wherein R ^A , R ^B and p are as previously defined herein.

またはその薬学的に許容可能な塩を有し、式中、Ｒ^ＡおよびＲ^Ｂは、本明細書に既に定義されるとおりである。 In embodiments, the cationic lipids of the present invention have the structure:

or a pharmaceutically acceptable salt thereof, wherein R ^A and R ^B are as previously defined herein.

またはその薬学的に許容可能な塩を有し、式中、Ｒ^ＡおよびＲ^Ｂは、本明細書に既に定義されるとおりである。 In embodiments, the cationic lipids of the present invention have the structure:

or a pharmaceutically acceptable salt thereof, wherein R ^A and R ^B are as previously defined herein.

またはその薬学的に許容可能な塩を有する。 In embodiments, the cationic lipids of the present invention have the structure:

or a pharmaceutically acceptable salt thereof.

またはその薬学的に許容可能な塩を有する。 In embodiments, the cationic lipids of the present invention have the structure:

or a pharmaceutically acceptable salt thereof.

またはその薬学的に許容可能な塩を含み、式中、Ｒ^’、Ｒ^６およびＸは、本明細書に既に定義されるとおりである。 In embodiments, the cationic lipid of the present invention is a compound having a structure according to formula (IIID):

or a pharmaceutically acceptable salt thereof, wherein R ^' , ^R6 and X are as previously defined herein.

またはその薬学的に許容可能な塩を含む。 In embodiments, the cationic lipids of the present invention are compounds having structures according to the following formulas (IIIE), (IIIF), (IIIG), (IIIH), (III), (IIIJ) or (IIIK):

or a pharmaceutically acceptable salt thereof.

In embodiments, the cationic lipids of the invention have the structure:

In embodiments, the cationic lipids of the invention have the structure:

In embodiments, the cationic lipids of the invention have the structure:

In embodiments, the cationic lipids of the invention have the structure:

In embodiments, the cationic lipids of the invention have the structure:

In embodiments, the cationic lipids of the invention have the structure:

In embodiments, the cationic lipids of the invention have the structure:

またはその薬学的に許容可能な塩を含み、式中、Ｒ^’、Ｒ^６およびＸは、本明細書に既に定義されるとおりである。 In embodiments, the cationic lipid of the present invention is a compound having a structure according to formula (IIIL):

or a pharmaceutically acceptable salt thereof, wherein R ^' , ^R6 and X are as previously defined herein.

式中、Ｍは、Ｈ、ＯＨ、ＯＭｅまたはＭｅから選択される化合物、
またはその薬学的に許容可能な塩を含み、式中、Ｒ^Ａ、Ｒ^Ｂ、ｍおよびｐは、本明細書に既に定義されるとおりである。 In embodiments, the cationic lipids of the present invention are compounds having a structure according to formula (IV):

wherein M is a compound selected from H, OH, OMe or Me;
or a pharmaceutically acceptable salt thereof, wherein R ^A , R ^B , m and p are as previously defined herein.

またはその薬学的に許容可能な塩を含み、
式中、ＹおよびＺのうちの一つはＯＨであり、他方は－ＯＣ（Ｏ）Ｒ’であり、またはＹおよびＺの両方がそれぞれ独立して、－ＯＣ（Ｏ）Ｒ’であり、式中、Ｒ^’、Ｒ^６およびＸは本明細書に既に定義されるとおりである。 In embodiments, the cationic lipid of the present invention is a compound having a structure according to formula (VI), (VII), (VIII), (IX) or (X) below,

or a pharmaceutically acceptable salt thereof,
wherein one of Y and Z is OH and the other is -OC(O)R', or both Y and Z are each independently -OC(O)R'; wherein R ^' , ^R6 and X are as previously defined herein.

In embodiments, one of Y and Z is OH and the other is -OC(O)R'.

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

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

In embodiments, both Y and Z are -OC(O)R'.

In embodiments, a composition comprising the cationic lipid of any one of the preceding embodiments, one or more non-cationic lipids, one or more cholesterol-based lipids, and one or more PEG-modified lipids. An article is provided herein. In embodiments, the composition is a lipid nanoparticle. In embodiments, one or more cationic lipids constitute about 30-60 mol % of the lipid nanoparticles. In embodiments, one or more non-cationic lipids constitute about 10-50 mol % of the lipid nanoparticles. In embodiments, one or more PEG-modified lipids constitute about 1-10 mol % of the lipid nanoparticles. In embodiments, cholesterol-based lipids constitute 10-50 mol % of the lipid nanoparticles. In embodiments, the lipid nanoparticles encapsulate nucleic acids, optionally mRNAs encoding peptides or proteins. In embodiments, the lipid nanoparticles have an encapsulation rate of mRNA of at least 70%. In embodiments, the lipid nanoparticles have an encapsulation rate of mRNA of at least 75%. In embodiments, the lipid nanoparticles have an encapsulation rate of mRNA of at least 80%. In embodiments, the lipid nanoparticles have an encapsulation rate of mRNA of at least 85%. In embodiments, the lipid nanoparticles have an encapsulation rate of mRNA of at least 90%. In embodiments, the lipid nanoparticles have an encapsulation rate of mRNA of at least 95%.

In embodiments, the composition of any one of the preceding embodiments is for use in therapy.

In embodiments, the composition of any one of the preceding embodiments is for use in a method of treating or preventing a disease amenable to treatment or prevention with a peptide or protein encoded by the mRNA. optionally the disease is (a) a protein deficiency, optionally the protein deficiency affects the liver, lung, brain or muscle, (b) an autoimmune disease, (c) an infection or (d) cancer.

In embodiments, the composition is administered by intravenous, intrathecal, or intramuscular, or pulmonary delivery, optionally via nebulization.

Exemplary Compounds Exemplary compounds include those listed in Tables 1-8.

Any of the compounds identified in Tables 1-8 above may be provided in the form of pharmaceutically acceptable salts and such salts are intended to be encompassed by the present invention.

Unless otherwise specified, R=C ₁₆ H ₂₉ has the following structure:

Unless otherwise specified, R=C ₁₆ H ₃₁ has the following structure:

The compounds of the invention described herein can be prepared according to methods known in the art, including the exemplary syntheses of the examples provided herein.

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

Synthesis of Nucleic Acids Nucleic acids according to the invention can be synthesized according to any known method. For example, mRNA according to the invention can be synthesized by in vitro transcription (IVT). Briefly, IVT typically comprises a promoter, a pool of ribonucleotide triphosphates, a buffer system that may include DTT and magnesium ions, and an appropriate RNA polymerase (e.g., T3, T7, mutated T7, or SP6 RNA polymerase), DNAse I, pyrophosphatase, and/or RNAse inhibitors. The exact conditions will depend on the particular application.

In some embodiments, DNA templates are transcribed in vitro for the preparation of mRNA according to the invention. A suitable DNA template typically has a promoter for in vitro transcription, such as the T3, T7, mutated T7, or SP6 promoter, followed by the desired nucleotide sequence and termination signals of the desired mRNA.

Desired mRNA sequences according to the invention can be determined and incorporated into a DNA template using standard methods. For example, starting from a desired amino acid sequence (eg, an enzyme sequence), a virtual back-translation is performed based on the degeneracy of the genetic code. An optimization algorithm can then be used for the selection of suitable codons. Typically, the G/C content can be optimized on the one hand to achieve the highest possible G/C content and on the other hand to maximize the frequency of tRNAs according to codon usage. The optimized RNA sequence can be established, displayed using, for example, a suitable display device, and compared to the original (wild-type) sequence. Secondary structure can also be analyzed to calculate the stabilizing and destabilizing properties or regions of the RNA, respectively.

modified mRNA
In some embodiments, mRNA according to the present invention can be synthesized as unmodified mRNA or modified mRNA. Modified mRNA contains nucleotide modifications in the RNA. Thus, modified mRNAs according to the present invention may contain nucleotide modifications, eg, backbone modifications, sugar modifications, or base modifications. In some embodiments, the mRNA contains, but is not limited to, purines (adenine (A), guanine (G)) or pyrimidines (thymine (T), cytosine (C), uracil (U)) naturally occurring can be synthesized from existing nucleotides and/or nucleotide analogues (modified nucleotides) and modified nucleotide analogues or derivatives of purines and pyrimidines such as 1-methyl-adenine, 2-methyl-adenine, 2-methylthio-N- 6-isopentenyl-adenine, N6-methyl-adenine, N6-isopentenyl-adenine, 2-thio-cytosine, 3-methyl-cytosine, 4-acetyl-cytosine, 5-methyl-cytosine, 2,6-diaminopurine , 1-methyl-guanine, 2-methyl-guanine, 2,2-dimethyl-guanine, 7-methyl-guanine, inosine, 1-methyl-inosine, pseudouracil (5-uracil), dihydro-uracil, 2-thio- Uracil, 4-thio-uracil, 5-carboxymethylaminomethyl-2-thio-uracil, 5-(carboxyhydroxymethyl)-uracil, 5-fluoro-uracil, 5-bromo-uracil, 5-carboxymethylaminomethyl- Uracil, 5-methyl-2-thio-uracil, 5-methyl-uracil, N-uracil-5-oxyacetic acid methyl ester, 5-methylaminomethyl-uracil, 5-methoxyaminomethyl-2-thio-uracil, 5 '-Methoxycarbonylmethyl-uracil, 5-methoxy-uracil, uracil-5-oxyacetic acid methyl ester, uracil-5-oxyacetic acid (v), 1-methyl-pseudouracil, cuocin, beta-D-mannosyl-cuocin, wybutoxin , and phosphoramidites, phosphorothioates, peptide nucleotides, methylphosphonates, 7-deazaguanosine, 5-methylcytosine, inosine, and the like. Preparation of such analogs is described, for example, in US Pat. No. 4,373,071, US Pat. No. 4,401,796, US Pat. Patent No. 4,500,707, U.S. Patent No. 4,668,777, U.S. Patent No. 4,973,679, U.S. Patent No. 5,047,524, U.S. Patent No. 5,132,418, United States No. 5,153,319, US Pat. No. 5,262,530, and US Pat. No. 5,700,642, the disclosures of which are incorporated by reference in their entireties.

Pharmaceutical Formulations of Cationic Lipids and Nucleic Acids In certain embodiments, the compounds of the invention described herein, as well as pharmaceutical and liposomal compositions comprising such lipids, contain encapsulated materials such as mRNA. can be used in formulations to facilitate delivery of one or more polynucleotides) to one or more target cells and subsequent transfection. For example, in certain embodiments, the cationic lipids described herein (and compositions, such as liposomal compositions comprising such lipids) are capable of receptor-mediated endocytosis, clathrin-mediated, and caveolae-mediated Endocytosis, phagocytosis, and macropinocytosis, fusogenicity, endosomal or lysosomal disruption, and/or any of the releasable properties that provide advantages of such compounds compared to other similarly classified lipids. It can be characterized as providing one or more.

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

As used herein, the terms "delivery vehicle," "import vehicle," "nanoparticle," or their grammatical equivalents are used interchangeably.

For example, the invention provides compositions (eg, pharmaceutical compositions) comprising a compound described herein and one or more polynucleotides. The composition (e.g., pharmaceutical composition) comprises one or more cationic lipids, one or more non-cationic lipids, one or more cholesterol-based lipids, and/or one or more PEG-modified lipids. can further include

In certain embodiments, the composition exhibits enhanced (eg, enhanced) ability to transfect one or more target cells. Accordingly, also provided herein are methods of transfecting one or more target cells. Such methods generally involve encapsulating one or more target cells with cationic lipids and/or pharmaceutical compositions disclosed herein (e.g., one or more polynucleotides described herein). with a liposomal formulation comprising a compound of (e.g., one or more polynucleotides) such that one or more target cells are transfected with the material (e.g., one or more polynucleotides) encapsulated therein. Become. As used herein, the term “transfection” or “transfection” refers to the transfection of one or more of the encapsulated materials (eg, nucleic acids and/or polynucleotides) into a cell, or preferably a target cell. refers to intracellular introduction into The introduced polynucleotide may be stable or transiently maintained in the target cell. The term "transfection efficiency" refers to the relative amount of such encapsulated material (e.g., polynucleotide) taken up by, introduced into, and/or expressed by a target cell to be transfected. . In practice, transfection efficiency can be estimated by the amount of reporter polynucleotide product produced by the target cell after transfection. In certain embodiments, the compounds and pharmaceutical compositions described herein exhibit high transfection efficiencies such that appropriate dosages of the encapsulated material (e.g., one or more polynucleotides) are It minimizes potential systemic side effects or toxicity associated with the compounds or their encapsulated contents while improving the likelihood of being delivered to and subsequently expressed at the site of disease.

For example, following transfection of one or more target cells with polynucleotides encapsulated in one or more lipid nanoparticles comprising the pharmaceutical or liposomal compositions disclosed herein, with such polynucleotides Production of the encoded product (e.g., polypeptide or protein) can preferably be stimulated to enhance the ability of such target cells to express the polynucleotide and, e.g., to produce the polypeptide or protein of interest. . For example, transfection of target cells with one or more compounds or pharmaceutical compositions that encapsulate mRNA enhances (ie, increases) production of the protein or enzyme encoded by such mRNA.

Additionally, the delivery vehicles (eg, liposomal delivery vehicles) described herein can be prepared to preferentially distribute to other target tissues, cells, or organs, such as heart, lung, kidney, spleen. . In embodiments, the lipid nanoparticles of the present invention can be prepared to achieve enhanced delivery to target cells and tissues. For example, polynucleotides (e.g., mRNA) encapsulated in one or more of the compounds or pharmaceutical compositions and liposomal compositions described herein can be delivered to and/or can be transfected. In some embodiments, the encapsulated polynucleotide (e.g., mRNA) is expressed and a functional polypeptide product is produced (and in some instances excreted) by the target cell, thereby imparting beneficial properties. , for example, to target cells or tissues. Such encapsulated polynucleotides (eg, mRNA) can encode, for example, hormones, enzymes, receptors, polypeptides, peptides, or other proteins of interest.

Liposomal Delivery Vehicles In some embodiments, the composition is a suitable delivery vehicle. In embodiments, the composition is a liposomal delivery vehicle, such as lipid nanoparticles.

The terms "liposome delivery vehicle" and "liposome composition" are used interchangeably.

Enrichment of the liposomal composition with one or more of the cationic lipids disclosed herein may be used as a means of ameliorating (e.g., reducing) toxicity or otherwise to one or more desired lipids. It can be used as a means of imparting properties to such concentrated liposomal compositions, such as improved delivery of the encapsulated polynucleotide to one or more target cells and/or reduced in vivo toxicity of the liposomal composition. . Accordingly, pharmaceutical compositions, particularly liposomal compositions, comprising one or more of the cationic lipids disclosed herein are also contemplated.

Thus, in certain embodiments, the compounds of the invention described herein effect the delivery and release of encapsulated materials (e.g., one or more therapeutic agents) to one or more target cells. To facilitate or enhance (eg, by penetrating or fusing with the lipid membranes of such target cells), they can be used as components of liposomal compositions.

As used herein, liposome delivery vehicles, e.g., lipid nanoparticles, are typically characterized as microscopic vesicles having an internal aqueous space separated from the external medium by one or more bilayer membranes. . The bilayer membrane of liposomes is typically formed by amphiphilic molecules, such as lipids of synthetic or natural origin, containing spatially separated hydrophilic and hydrophobic domains (Lasic, Trends Biotechnol. , 16:307-321, 1998). The bilayer membrane of liposomes may also be formed by amphipathic polymers and surfactants (eg, polymerosomes, niosomes, etc.). In the context of the present invention, liposomal delivery vehicles typically serve to transport desired mRNA to target cells or tissues.

In certain embodiments, such compositions (e.g., liposomal compositions) are loaded with or contain materials such as, for example, one or more biologically active polynucleotides (e.g., mRNA). Enclosed.

In embodiments, a composition (eg, a pharmaceutical composition) comprises mRNA encoding a protein encapsulated within liposomes. In embodiments, the liposomes comprise one or more cationic lipids, one or more non-cationic lipids, one or more cholesterol-based lipids, and one or more PEG-modified lipids, and at least One cationic lipid is a compound of the invention described herein. In embodiments, the composition comprises mRNA encoding a protein (eg, any protein described herein). In embodiments, the composition comprises mRNA encoding cystic fibrosis transmembrane conductance regulator (CFTR) protein. In embodiments, the composition comprises mRNA encoding ornithine transcarbamylase (OTC) protein.

In embodiments, a composition (eg, a pharmaceutical composition) comprises a nucleic acid encapsulated within a liposome, and the liposome comprises a compound described herein.

In embodiments, the nucleic acid is mRNA encoding a peptide or protein. In embodiments, the mRNA encodes a peptide or protein for use in delivery to or treatment of the lung or lung cells of a subject (e.g., the mRNA is the cystic fibrosis transmembrane conductance regulator (CFTR) protein code). In embodiments, the mRNA encodes a peptide or protein for use in delivery to or treatment of the liver or liver cells of a subject (e.g., the mRNA encodes an ornithine transcarbamylase (OTC) protein). . Still other exemplary mRNAs are described herein.

In embodiments, liposomal delivery vehicles (eg, lipid nanoparticles) can have a net positive charge.

In embodiments, liposomal delivery vehicles (eg, lipid nanoparticles) can have a net negative charge.

In embodiments, liposomal delivery vehicles (eg, lipid nanoparticles) can have a net neutral charge.

In embodiments, lipid nanoparticles encapsulating nucleic acids (eg, mRNA encoding peptides or proteins) comprise one or more compounds of the invention described herein.

For example, the amount of a compound of the invention described herein in a composition is a percentage of the total dry weight of all lipids in the composition (e.g., the total dry weight of all lipids present in the liposomal composition) ("% by weight").

In some embodiments of the pharmaceutical compositions described herein, the compound of the invention described herein is less than the total dry weight of all lipids present in the composition (e.g., liposomal composition). It is present in an amount of about 0.5% to about 30% (eg, about 0.5% to about 20%) by weight.

In embodiments, the compounds of the invention described herein comprise from about 1% to about 30% by weight of the total dry weight of all lipids present in the composition (eg, liposomal composition), about It is present in an amount of 1% to about 20%, about 1% to about 15%, about 1% to about 10%, or about 5% to about 25% by weight. In embodiments, the compounds of the invention described herein comprise from about 0.5% to about 5% by weight of the total dry weight of all lipids present in the composition, such as a liposomal delivery vehicle, about It is present in an amount of 1% to about 10%, about 5% to about 20%, or about 10% to about 20% by weight.

In embodiments, the amount of a compound of the invention described herein is at least about 5%, about 10%, about 15 wt%, about 20 wt%, about 25 wt%, about 30 wt%, about 35 wt%, about 40 wt%, about 45 wt%, about 50 wt%, about 55 wt%, about 60 wt%, about 65% by weight, about 70% by weight, about 75% by weight, about 80% by weight, about 85% by weight, about 90% by weight, about 95% by weight, about 96% by weight, about 97% by weight, about 98% by weight, or It is present in an amount of about 99% by weight.

In embodiments, the amount of a compound of the invention described herein is no greater than about 5%, no greater than about 10%, no greater than about 10% by weight of the total dry weight of total lipids in the composition (e.g., liposomal composition); about 15 wt% or less, about 20 wt% or less, about 25 wt% or less, about 30 wt% or less, about 35 wt% or less, about 40 wt% or less, about 45 wt% or less, about 50 wt% or less, about 55 wt% wt% or less, about 60 wt% or less, about 65 wt% or less, about 70 wt% or less, about 75 wt% or less, about 80 wt% or less, about 85 wt% or less, about 90 wt% or less, about 95 wt% less than or equal to about 96 weight percent, less than or equal to about 97 weight percent, less than or equal to about 98 weight percent, or less than or equal to about 99 weight percent.

In embodiments, the composition (eg, a liposome delivery vehicle such as a lipid nanoparticle) comprises from about 0.1% to about 20% (eg, from about 0.1% to about 15%) of the present Including the compounds described in the specification. In embodiments, the delivery vehicle (e.g., a liposome delivery vehicle such as a lipid nanoparticle) comprises about 0.5 wt%, about 1 wt%, about 3 wt%, about 5 wt%, or about 10 wt% Including compounds described herein. In embodiments, the delivery vehicle (e.g., a liposome delivery vehicle such as a lipid nanoparticle) is up to about 0.5%, up to about 1%, up to about 3%, up to about 5%, up to about 10%, up to about 15%, or up to about 20% by weight of a compound described herein. In embodiments, this percentage results in an improved beneficial effect (eg, improved delivery to target tissues such as the liver or lungs).

The amount of a compound of the invention described herein in a composition may also be a percentage of the total molar amount of total lipids in the composition (e.g., the total molar amount of all lipids present in the liposome delivery vehicle) (" mol %”).

In some embodiments of the pharmaceutical compositions described herein, the compounds of the invention described herein are about 0 of the total molar amount of all lipids present in the composition, such as the liposomal delivery vehicle. .5 mol % to about 50 mol % (eg, about 0.5 mol % to about 20 mol %).

In embodiments, the compounds of the invention described herein comprise about 0.5 mol% to about 5 mol% of the total molar amount of all lipids present in the composition, such as a liposomal delivery vehicle, about 1 mol% to about 10 mol%, about 5 mol% to about 20 mol%, about 10 mol% to about 20 mol%, about 15 mol% to about 30 mol%, about 20 mol% to about 35 mol%, about 25 mol% to about 40 mol%, about 30 mol% to about 45 mol%, about 35 mol% to about 50 mol%, about 40 mol% to about 55 mol%, or about 45 mol% to about 60 mol% present in quantity. In embodiments, the compounds of the invention described herein are from about 1 mol% to about 60 mol% of the total molar amount of all lipids present in the composition, such as a liposomal delivery vehicle, about 1 mol. % to about 50 mol %, about 1 mol % to about 40 mol %, about 1 mol % to about 30 mol %, about 1 mol % to about 20 mol %, about 1 mol % to about 15 mol %, about 1 mol % to about 10 mol %, about 5 mol % to about 55 mol %, about 5 mol % to about 45 mol %, about 5 mol % to about 35 mol %, or about 5 mol % to about 25 mol % exist.

In certain embodiments, the compounds of the invention described herein are present at about 0.1 mol % to about 50 mol %, or 0.5 mol, of the total amount of lipids in the composition (eg, liposomal delivery vehicle). % to about 50 mol %, or about 1 mol % to about 25 mol %, or about 1 mol % to about 10 mol %.

In certain embodiments, the compounds of the invention described herein are greater than about 0.1 mol%, or greater than about 0.5 mol%, or greater than about 1 mol% of the total amount of lipids in the lipid nanoparticles. , greater than about 5 mol %, greater than about 10 mol %, greater than about 20 mol %, greater than about 30 mol %, or greater than about 40 mol %.

In certain embodiments, the described compounds are less than about 60 mol%, or less than about 55 mol%, or less than about 50 mol%, or less than about 45 mol% of the total amount of lipids in the composition (e.g., liposomal delivery vehicle). less than about 40 mol%, or less than about 35 mol%, less than about 30 mol%, or less than about 25 mol%, or less than about 10 mol%, or less than about 5 mol%, or about 1 mol% can include less than

In embodiments, the amount of a compound of the invention described herein is at least about 5 mol %, about 10 mol %, about 15 mol%, about 20 mol%, about 25 mol%, about 30 mol%, about 35 mol%, about 40 mol%, about 45 mol%, about 50 mol%, about 55 mol%, about 60 mol%, about 65 mol%, about 70 mol%, about 75 mol%, about 80 mol%, about 85 mol%, about 90 mol%, about 95 mol%, about 96 mol%, about 97 mol%, about 98 mol%, or It is present in an amount of about 99 mole %.

In embodiments, the amount of the compounds of the invention described herein is no more than about 5 mol%, no more than about 10 mol% of the total molar amount of total lipids in the composition (e.g., liposomal composition), about 15 mol% or less, about 20 mol% or less, about 25 mol% or less, about 30 mol% or less, about 35 mol% or less, about 40 mol% or less, about 45 mol% or less, about 50 mol% or less, about 55 mol % or less, about 60 mol % or less, about 65 mol % or less, about 70 mol % or less, about 75 mol % or less, about 80 mol % or less, about 85 mol % or less, about 90 mol % or less, about 95 mol % less than or equal to about 96 mol %, less than or equal to about 97 mol %, less than or equal to about 98 mol %, or less than or equal to about 99 mol %.

In embodiments, this percentage results in an improved beneficial effect (eg, improved delivery to target tissues such as the liver or lungs).

In an exemplary embodiment, the compositions (e.g., liposomal compositions) of the invention comprise one or more cationic lipids, one or more non-cationic lipids, one or more cholesterol-based lipids, and One or more PEG-modified lipids, wherein at least one cationic lipid is a compound of the invention as described herein. For example, a composition suitable for practicing the present invention comprises four lipid components, including a compound of the invention described herein as the cationic lipid component, a non-cationic lipid, a cholesterol-based lipid, and a PEG-modified lipid. have. The non-cationic lipid may be DOPE or DEPE. The cholesterol-based lipid may be cholesterol. The PEG-modified lipid may be DMG-PEG2K.

In embodiments, a composition of the invention comprises a cationic lipid of the invention, DMG-PEG2000, cholesterol and DOPE, wherein the molar ratio of cationic lipid:DMG-PEG2000:cholesterol:DOPE is 40:5:25. :30.

In further embodiments, the pharmaceutical (eg, liposomal) composition comprises one or more of PEG-modified lipids, non-cationic lipids, and cholesterol lipids. In other embodiments, such pharmaceutical (eg, liposome) compositions comprise one or more PEG-modified lipids, one or more non-cationic lipids, and one or more cholesterol lipids. In still further embodiments, such pharmaceutical (eg, liposome) compositions comprise one or more PEG-modified lipids and one or more cholesterol lipids.

In embodiments, compositions (e.g., lipid nanoparticles) encapsulating nucleic acids (e.g., mRNA encoding peptides or proteins) comprise one or more compounds of the invention described herein and a cation. one or more lipids selected from the group consisting of cationic lipids, non-cationic lipids, and pegylated lipids.

In embodiments, compositions (e.g., lipid nanoparticles) encapsulating nucleic acids (e.g., mRNA encoding peptides or proteins) contain one or more compounds of the invention described herein; comprising one or more lipids selected from the group consisting of lipids, non-cationic lipids, and pegylated lipids; and further comprising cholesterol-based lipids. Typically, such compositions will contain a compound of the invention described herein as the cationic lipid component, non-cationic lipids (e.g. DOPE), cholesterol-based lipids (e.g. cholesterol), and PEG-modified lipids (e.g. cholesterol). For example, it has four lipid components including DMG-PEG2K).

In embodiments, a lipid nanoparticle encapsulating a nucleic acid (e.g., mRNA encoding a peptide or protein) comprises one or more compounds of the invention described herein as well as cationic lipids, non-cationic It comprises one or more lipids selected from the group consisting of lipids, pegylated lipids, and cholesterol-based lipids.

According to various embodiments, the selection of cationic lipids, non-cationic lipids, and/or PEG-modified lipids, including lipid nanoparticles, and the selection of relative molar proportions of such lipids to each other is a characteristic of the selected lipids. , the nature of the target cell of interest, and the characteristics of the mRNA to be delivered. Additional considerations include, for example, the degree of saturation of the alkyl chain of the lipid chosen, as well as size, charge, pH, pKa, fusogenicity, and toxicity. Therefore, the molar ratio can be adjusted accordingly.

In some embodiments, the ratio of cationic lipid: non-cationic lipid: cholesterol-based lipid: PEG-modified lipid can be about 30-60:20-40:20-30:1-10, respectively. In some embodiments, the ratio of cationic lipid to non-cationic lipid to cholesterol-based lipid to PEG-modified lipid is approximately 40:30:20:10, respectively. In some embodiments, the ratio of cationic lipids to non-cationic lipids to cholesterol-based lipids to PEG-modified lipids is approximately 40:30:25:5, respectively. In some embodiments, the ratio of cationic lipids to non-cationic lipids to cholesterol-based lipids to PEG-modified lipids is approximately 40:32:25:3, respectively. In some embodiments, the ratio of cationic lipid: non-cationic lipid: cholesterol-based lipid: PEG-modified lipid is approximately 50:25:20:5.

Cationic Lipids In addition to any of the compounds of the invention described herein, the compositions may contain one or more additional cationic lipids.

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

Additional cationic lipids suitable for use in the present compositions include the cationic lipids described in this document.

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

In some embodiments, the non-cationic lipid is a neutral lipid, ie, a lipid that has no net charge under the conditions under which the composition is formulated and/or administered.

In some embodiments, non-cationic lipids are from about 5% to about 90%, from about 5% to about 70%, from about 5% to about 50%, from about 5% to about 90% of the total lipids present in the composition. It can be present in a molar ratio (mole %) of about 40%, about 5% to about 30%, about 10% to about 70%, about 10% to about 50%, or about 10% to about 40%. In some embodiments, total non-cationic lipids are about 5% to about 90%, about 5% to about 70%, about 5% to about 50%, about 5% of the total lipids present in the composition It can be present in a molar ratio (mole %) of from about 40%, from about 5% to about 30%, from about 10% to about 70%, from about 10% to about 50%, or from about 10% to about 40%. In some embodiments, the percentage of non-cationic lipids in the liposomes is greater than about 5 mol%, greater than about 10 mol%, greater than about 20 mol%, greater than about 30 mol%, or greater than about 40 mol%. obtain. In some embodiments, the percentage of total non-cationic lipids in the liposomes is greater than about 5 mol%, greater than about 10 mol%, greater than about 20 mol%, greater than about 30 mol%, or greater than about 40 mol%. could be. In some embodiments, the percentage of non-cationic lipids in the liposome is no greater than about 5 mol%, no greater than about 10 mol%, no greater than about 20 mol%, no greater than about 30 mol%, or no greater than about 40 mol%. . In some embodiments, the percentage of total non-cationic lipids in the liposomes is no more than about 5 mol%, no more than about 10 mol%, no more than about 20 mol%, no more than about 30 mol%, or no more than about 40 mol%. could be.

In some embodiments, the non-cationic lipid is about 5% to about 90%, about 5% to about 70%, about 5% to about 50%, about 5% to about 90%, about 5% to about 50%, about 5% of the total composition lipids present in the composition. % to about 40%, about 5% to about 30%, about 10% to about 70%, about 10% to about 50%, or about 10% to about 40%. In some embodiments, total non-cationic lipids are about 5% to about 90%, about 5% to about 70%, about 5% to about 50%, about 5% of the total lipids present in the composition It can be present in a weight ratio (wt %) of from about 40%, from about 5% to about 30%, from about 10% to about 70%, from about 10% to about 50%, or from about 10% to about 40%. In some embodiments, the percentage of non-cationic lipids in the liposomes is greater than about 5%, greater than about 10%, greater than about 20%, greater than about 30%, or greater than about 40% by weight. obtain. In some embodiments, the percentage of total non-cationic lipids in the liposomes is greater than about 5%, greater than about 10%, greater than about 20%, greater than about 30%, or greater than about 40% by weight. could be. In some embodiments, the percentage of non-cationic lipids in the liposomes is no more than about 5%, no more than about 10%, no more than about 20%, no more than about 30%, or no more than about 40% by weight. . In some embodiments, the percentage of total non-cationic lipids in the liposomes is no more than about 5%, no more than about 10%, no more than about 20%, no more than about 30%, or no more than about 40% by weight. could be.

Cholesterol-Based Lipids In some embodiments, compositions (eg, liposomal compositions) comprise one or more cholesterol-based lipids. For example, a preferred cholesterol-based lipid for practicing the invention is cholesterol. Other suitable cholesterol-based lipids include, for example, DC-Chol (N,N-dimethyl-N-ethylcarboxamidocholesterol), 1,4-bis(3-N-oleylamino-propyl)piperazine (Gao, et al. Biochem.Biophys.Res.Comm.179,280 (1991), Wolf et al.BioTechniques 23,139 (1997), U.S. Patent No. 5,744,335), or imidazole cholesterol ester (ICE )including.

In some embodiments, cholesterol-based lipids can be present in a molar ratio (mole %) of about 1% to about 30%, or about 5% to about 20%, of the total lipids present in the liposome. In some embodiments, the percentage of cholesterol-based lipids in the lipid nanoparticles is greater than about 5 mol%, greater than about 10 mol%, greater than about 20 mol%, greater than about 30 mol%, or greater than about 40 mol%. could be. In some embodiments, the percentage of cholesterol-based lipids in the lipid nanoparticles is no more than about 5 mol%, no more than about 10 mol%, no more than about 20 mol%, no more than about 30 mol%, or no more than about 40 mol%. could be.

In some embodiments, the cholesterol-based lipid can be present in a weight ratio (wt %) of about 1% to about 30%, or about 5% to about 20%, of the total lipids present in the liposome. In some embodiments, the percentage of cholesterol-based lipids in the lipid nanoparticles is greater than about 5%, greater than about 10%, greater than about 20%, greater than about 30%, or greater than about 40% by weight. could be. In some embodiments, the percentage of cholesterol-based lipids in the lipid nanoparticles is no more than about 5%, no more than about 10%, no more than about 20%, no more than about 30%, or no more than about 40% by weight. could be.

Pegylated Lipids In some embodiments, the composition (eg, liposomal composition) comprises one or more additional pegylated lipids. A preferred PEG-modified or PEGylated lipid for practicing the present invention is 1,2-dimyristoyl-rac-glycero-3-methoxypolyethyleneglycol-2000 (DMG-PEG2K).

For example, polyethylene glycol (PEG)-modified phospholipids and derivatized ceramides (PEG- CER) is also contemplated by the present invention in combination with one or more compounds of the invention described herein, and in some embodiments, together with other lipids, including liposomes. In some embodiments, a particularly useful exchangeable lipid is PEG-ceramide with shorter acyl chains (eg, C ₁₄ or C ₁₈ ).

Additional contemplated PEG-modified lipids (also referred to herein as PEGylated lipids, this term being interchangeable with PEG-modified lipids) include covalently attached lipids having alkyl chains of _C6 - _C20 length. including, but not limited to, polyethylene glycol chains up to 5 kDa in length. In some embodiments, the PEG-modified or PEGylated lipid is PEGylated cholesterol or PEG-2K. Addition of such moieties can prevent aggregation of complexes, increase circulation lifespan, and can also provide a means for increasing delivery of lipid-nucleic acid compositions to target cells (Klibanov et al. (1990) FEBS Letters, 268(1):235-237) or these components can be selected for rapid exchange from the formulation in vivo (see US Pat. No. 5,885,613). ).

Additional PEG-modified phospholipids and derivatized lipids of the present invention may be from about 0% to about 10%, from about 0.5% to about 10%, from about 1 % to about 10%, about 2% to about 10%, or about 3% to about 5%.

Pharmaceutical Formulations and Therapeutic Uses The compounds of the invention described herein facilitate the delivery and release of encapsulated materials (eg, one or more therapeutic polynucleotides) to one or more target cells. or enhance (eg, by penetrating or fusing with the lipid membranes of such target cells), may be used in the preparation of compositions (eg, constructing liposomal compositions).

For example, when the liposomal composition (e.g., lipid nanoparticles) comprises or is otherwise enriched with one or more of the compounds disclosed herein, one or more target cells A phase transition in the lipid bilayer of a can facilitate delivery of encapsulated materials (e.g., one or more therapeutic polynucleotides encapsulated in lipid nanoparticles) to one or more target cells.

Similarly, in certain embodiments, the compounds of the invention described herein can be used to prepare liposomal vehicles that are characterized by reduced toxicity in vivo. In certain embodiments, reduced toxicity is associated with the compositions disclosed herein such that reduced amounts of such compositions can be administered to a subject to achieve a desired therapeutic response or outcome. It is a function of the associated high transfection efficiency.

Accordingly, pharmaceutical formulations containing the compounds described and the nucleic acids provided by the invention can be used for a variety of therapeutic purposes. To facilitate delivery of nucleic acids in vivo, the compounds and nucleic acids described herein are formulated in combination with one or more additional pharmaceutical carriers, targeting ligands, or stabilizing reagents. good too. In some embodiments, the compounds described herein can be formulated via premixed lipid solutions. In other embodiments, compositions containing compounds described herein can be formulated using techniques for post-insertion of nanoparticles into lipid membranes. Formulation techniques and drug administration are described in "Remington's Pharmaceutical Sciences," Mack Publishing Co.; , Easton, Pa. (latest version).

Suitable routes of administration include, for example, pulmonary administration including oral, rectal, vaginal, transmucosal, intratracheal or inhaled administration, or enteral administration, intradermal injection, transdermal (topical) injection, intramuscular Included are parenteral delivery, including injection, subcutaneous injection, intramedullary injection, as well as intrathecal, direct intracerebroventricular, intravenous, intraperitoneal, or intranasal. In certain embodiments, intramuscular administration is to muscle selected from the group consisting of skeletal muscle, smooth muscle, and cardiac muscle. In some embodiments, this administration results in delivery of the nucleic acid to muscle cells. In some embodiments, this administration results in delivery of the nucleic acid to hepatocytes (ie liver cells).

A common route for administering the liposomal compositions of the invention has been intravenous delivery, particularly when treating metabolic disorders, particularly those affecting the liver (e.g., ornithine transcarbamylase (OTC) deficiency). may Alternatively, depending on the disease or disorder to be treated, liposomal compositions can be administered via pulmonary delivery (eg, for treatment of cystic fibrosis). For vaccination, the liposomal compositions of the invention are typically administered intramuscularly. Diseases or disorders affecting the eye can be treated by intravitreal administration of the liposomal compositions of the present invention.

Alternatively or additionally, the pharmaceutical formulations of the present invention may be administered in a local rather than systemic manner, for example, by direct injection of the pharmaceutical formulation into the targeted tissue, preferably in a sustained release formulation. Local delivery can be accomplished in a variety of ways depending on the tissue targeted. Exemplary tissues in which the delivered mRNA can be delivered and/or expressed include liver, kidney, heart, spleen, serum, brain, skeletal muscle, lymph nodes, skin, and/or cerebrospinal fluid, It is not limited to these. In embodiments, the targeted tissue is the liver. For example, an aerosol containing a composition of the invention can be inhaled (for nasal, tracheal, or bronchial delivery), and a composition of the invention can be injected, for example, at the site of injury, disease manifestation, or pain. , the composition may be provided in oral, tracheal, or esophageal lozenges; may be supplied in liquid, tablet, or capsule form for gastric or intestinal administration; or may be supplied in suppository form for rectal or vaginal administration. or it can also be delivered to the eye by using creams, drops, or even injections.

The compositions described herein can include mRNA encoding peptides, including peptides (eg, polypeptides such as proteins) described herein.

In some embodiments, the mRNA encodes the polypeptide.

In some embodiments, the mRNA encodes a protein.

Examples of peptides encoded by mRNA (eg, proteins encoded by mRNA) are described herein.

The present invention provides compositions having full-length mRNA molecules encoding peptides or proteins for use in the treatment of subjects, e.g., human subjects, or cells of human subjects, or cells to be treated and delivered to human subjects. Provide a method for delivery.

Thus, in certain embodiments, the present invention provides methods for producing therapeutic compositions comprising full-length mRNA encoding peptides or proteins for delivery to, or use in treatment of, the lungs or lung cells of a subject. offer. In certain embodiments, the invention provides methods for producing therapeutic compositions having full-length mRNA encoding cystic fibrosis transmembrane conductance regulator (CFTR) protein. In certain embodiments, the invention provides methods for producing therapeutic compositions having full-length mRNA encoding ATP-binding cassette subfamily A member 3 proteins. In certain embodiments, the present invention provides production for producing therapeutic compositions having full-length mRNA encoding dynein axonem intermediate strand 1 protein. In certain embodiments, the invention provides methods for producing therapeutic compositions having full-length mRNA encoding dynein axoneme heavy chain 5 (DNAH5) protein. In certain embodiments, the invention provides methods for producing therapeutic compositions having full-length mRNA encoding alpha-1-antitrypsin protein. In certain embodiments, the invention provides methods for producing therapeutic compositions having full-length mRNA encoding forkhead box P3 (FOXP3) protein. In certain embodiments, the invention has full-length mRNAs encoding one or more surfactant proteins, e.g., one or more of Surfactant A protein, Surfactant B protein, Surfactant C protein, and Surfactant D protein. Methods are provided for producing therapeutic compositions.

In certain embodiments, the present invention provides methods for producing therapeutic compositions having full-length mRNA encoding peptides or proteins for use in delivery to or treatment of the liver or liver cells of a subject. . Such peptides and polypeptides are associated with urea cycle disorders, lysosomal storage disorders, glycogen storage disorders, amino acid metabolism disorders, lipid metabolism or fibrotic disorders, Those associated with methylmalonic acidemia, or any other metabolic disorder for which delivery of or treatment with enriched full-length mRNA to the liver or liver cells would provide a therapeutic benefit.

In certain embodiments, the invention provides methods for producing therapeutic compositions having full-length mRNAs encoding proteins associated with urea cycle disorders. In certain embodiments, the invention provides methods for producing therapeutic compositions having full-length mRNA encoding ornithine transcarbamylase (OTC) protein. In certain embodiments, the invention provides methods for producing therapeutic compositions having full-length mRNA encoding argininosuccinate synthetase 1 protein. In certain embodiments, the invention provides methods for producing therapeutic compositions having full-length mRNA encoding carbamoyl phosphate synthetase I protein. In certain embodiments, the invention provides methods for producing therapeutic compositions having full-length mRNA encoding an argininosuccinate lyase protein. In certain embodiments, the invention provides methods for producing therapeutic compositions having full-length mRNA encoding an arginase protein.

In certain embodiments, the invention provides methods for producing therapeutic compositions having full-length mRNAs encoding proteins associated with lysosomal storage disorders. In certain embodiments, the invention provides methods for producing therapeutic compositions having full-length mRNA encoding an alpha-galactosidase protein. In certain embodiments, the invention provides methods for producing therapeutic compositions having full-length mRNA encoding a glucocerebrosidase protein. In certain embodiments, the invention provides methods for producing therapeutic compositions having full-length mRNA encoding iduronate-2-sulfatase protein. In certain embodiments, the invention provides methods for producing therapeutic compositions having full-length mRNA encoding iduronidase protein. In certain embodiments, the invention provides methods for producing therapeutic compositions having full-length mRNA encoding N-acetyl-alpha-D-glucosaminidase protein. In certain embodiments, the invention provides methods for producing therapeutic compositions having full-length mRNA encoding heparan N-sulfatase protein. In certain embodiments, the invention provides methods for producing therapeutic compositions having full-length mRNA encoding galactosamine-6 sulfatase protein. In certain embodiments, the invention provides methods for producing therapeutic compositions having full-length mRNA encoding beta-galactosidase protein. In certain embodiments, the invention provides methods for producing therapeutic compositions having full-length mRNA encoding a lysosomal lipase protein. In certain embodiments, the invention provides methods for producing therapeutic compositions having full-length mRNA encoding arylsulfatase B (N-acetylgalactosamine-4-sulfatase) protein. In certain embodiments, the invention provides methods for producing therapeutic compositions having full-length mRNA encoding transcription factor EB (TFEB).

In certain embodiments, the invention provides methods for producing therapeutic compositions having full-length mRNAs encoding proteins associated with glycogen storage disorders. In certain embodiments, the invention provides methods for producing therapeutic compositions having full-length mRNA encoding acid alpha-glucosidase protein. In certain embodiments, the invention provides methods for producing therapeutic compositions having full-length mRNA encoding glucose-6-phosphatase (G6PC) protein. In certain embodiments, the invention provides methods for producing therapeutic compositions having full-length mRNA encoding liver glycogen phosphorylase protein. In certain embodiments, the invention provides methods for producing therapeutic compositions having full-length mRNA encoding muscle phosphoglycerate mutase protein. In certain embodiments, the invention provides methods for producing therapeutic compositions having full-length mRNA encoding glycogen debranching enzyme.

In certain embodiments, the invention provides methods for producing therapeutic compositions having full-length mRNAs encoding proteins related to amino acid metabolism. In certain embodiments, the invention provides methods for producing therapeutic compositions having full-length mRNA encoding a phenylalanine hydroxylase enzyme. In certain embodiments, the invention provides methods for producing therapeutic compositions having full-length mRNA encoding the glutaryl-CoA dehydrogenase enzyme. In certain embodiments, the invention provides methods for producing therapeutic compositions having full-length mRNA encoding a propionyl-CoA carboxylase enzyme. In certain embodiments, the invention provides methods for producing therapeutic compositions having full-length mRNA encoding the oxalase alanine-glyoxyl aminotransferase enzyme.

In certain embodiments, the invention provides methods for producing therapeutic compositions having full-length mRNAs encoding proteins associated with lipid metabolism or fibrotic disorders. In certain embodiments, the invention provides methods for producing therapeutic compositions having full-length mRNA encoding mTOR inhibitors. In certain embodiments, the invention provides methods for producing therapeutic compositions having full-length mRNA encoding ATPase phospholipid transport 8B1 (ATP8B1) protein. In certain embodiments, the invention provides one or more NF-kappaB inhibitors, such as one of I-kappaB alpha, interferon-related developmental regulator 1 (IFRD1), and sirtuin 1 (SIRT1). Methods are provided for producing therapeutic compositions having full-length mRNAs encoding one or more. In certain embodiments, the invention provides methods for producing therapeutic compositions having full-length mRNA encoding a PPAR-gamma protein or activity variant.

In certain embodiments, the invention provides methods for producing therapeutic compositions having full-length mRNA encoding the methylmalonic acidemia associated protein. For example, in certain embodiments, the invention provides methods for producing therapeutic compositions having full-length mRNA encoding methylmalonyl-CoA mutase protein. In certain embodiments, the invention provides methods for producing therapeutic compositions having full-length mRNA encoding a methylmalonyl-CoA epimerase protein.

In certain embodiments, the present invention provides methods for producing therapeutic compositions having full-length mRNA whose delivery to the liver or treatment thereof can provide therapeutic benefit. In certain embodiments, the present invention provides methods for producing therapeutic compositions having full-length mRNA encoding ATP7B protein, aka Wilson's disease protein. In certain embodiments, the invention provides methods for producing therapeutic compositions having full-length mRNA encoding a porphobilinogen deaminase enzyme. In certain embodiments, the invention provides therapeutic compositions having full-length mRNA encoding one or more clotting enzymes, such as Factor VIII, Factor IX, Factor VII, and Factor X. provide a method of In certain embodiments, the invention provides methods for producing therapeutic compositions having full-length mRNA encoding human hemochromatosis (HFE) protein.

In certain embodiments, the invention provides methods for producing therapeutic compositions having full-length mRNA encoding peptides or proteins for use in delivery to or treatment of cardiovascular structures or cells in a subject. I will provide a. In certain embodiments, the invention provides methods for producing therapeutic compositions having full-length mRNA encoding vascular endothelial growth factor A protein. In certain embodiments, the invention provides methods for producing therapeutic compositions having full-length mRNA encoding a relaxin protein. In certain embodiments, the invention provides methods for producing therapeutic compositions having full-length mRNA encoding bone morphogenetic protein-9 protein. In certain embodiments, the invention provides methods for producing therapeutic compositions having full-length mRNA encoding bone morphogenetic protein-2 receptor protein.

In certain embodiments, the present invention provides methods for producing therapeutic compositions having full-length mRNA encoding peptides or proteins for use in delivery to or treatment of muscles or muscle cells of a subject. . In certain embodiments, the invention provides methods for producing therapeutic compositions having full-length mRNA encoding a dystrophin protein. In certain embodiments, the invention provides methods for producing therapeutic compositions having full-length mRNA encoding a frataxin protein. In certain embodiments, the present invention provides methods for producing therapeutic compositions having full-length mRNA encoding peptides or proteins for use in delivery to or treatment of the myocardium or myocardial cells of a subject. . In certain embodiments, the invention provides methods for producing therapeutic compositions having full-length mRNAs encoding proteins that modulate one or both of potassium and sodium channels in muscle tissue or muscle cells. In certain embodiments, the invention provides methods for producing therapeutic compositions having full-length mRNAs encoding proteins that modulate Kv7.1 channels in muscle tissue or muscle cells. In certain embodiments, the invention provides methods for producing therapeutic compositions having full-length mRNAs encoding proteins that modulate Nav1.5 channels in muscle tissue or muscle cells.

In certain embodiments, the present invention provides methods for producing therapeutic compositions having full-length mRNA encoding peptides or proteins for use in delivery to or treatment of the nervous system or nervous system cells of a subject. offer. For example, in certain embodiments, the invention provides methods for producing therapeutic compositions having full-length mRNA encoding Survival Motor Neuron 1 protein. For example, in certain embodiments, the invention provides methods for producing therapeutic compositions having full-length mRNA encoding survival motor neuron 2 protein. In certain embodiments, the invention provides methods for producing therapeutic compositions having full-length mRNA encoding a frataxin protein. In certain embodiments, the invention provides methods for producing therapeutic compositions having full-length mRNA encoding ATP-binding cassette subfamily D member 1 (ABCD1) proteins. In certain embodiments, the invention provides methods for producing therapeutic compositions having full-length mRNA encoding CLN3 protein.

In certain embodiments, the present invention produces therapeutic compositions having full-length mRNA encoding peptides or proteins for use in the blood or bone marrow, or blood or bone marrow cells, of a subject for delivery to or treatment thereof. provide a method for In certain embodiments, the invention provides methods for producing therapeutic compositions having full-length mRNA encoding beta-globin protein. In certain embodiments, the invention provides methods for producing therapeutic compositions having full-length mRNA encoding Bruton's tyrosine kinase protein. In certain embodiments, the invention produces therapeutic compositions having full-length mRNAs encoding one or more clotting enzymes, such as Factor VIII, Factor IX, Factor VII, and Factor X. provide a method for

In certain embodiments, the present invention provides methods for producing therapeutic compositions having full-length mRNA encoding peptides or proteins for use in delivery to or treatment of the kidney or kidney cells of a subject. . In certain embodiments, the present invention provides methods for producing therapeutic compositions having full-length mRNA encoding type IV collagen alpha 5 chain (COL4A5) protein.

In certain embodiments, the present invention provides methods for producing therapeutic compositions having full-length mRNA encoding peptides or proteins for use in delivery to or treatment of the eye or ocular cells of a subject. . In certain embodiments, the invention provides methods for producing therapeutic compositions having full-length mRNA encoding ATP-binding cassette subfamily A member 4 (ABCA4) protein. In certain embodiments, the invention provides methods for producing therapeutic compositions having full-length mRNA encoding retinoschisin protein. In certain embodiments, the present invention provides methods for producing therapeutic compositions having full-length mRNA encoding retinal pigment epithelium-specific 65 kDa (RPE65) protein. In certain embodiments, the invention provides methods for producing therapeutic compositions having full-length mRNA encoding the 290 kDa centrosomal protein (CEP290).

In certain embodiments, the invention provides methods for producing therapeutic compositions having full-length mRNA encoding peptides or proteins for use in delivery of vaccines or treatment with vaccines to a subject or cells of a subject. I will provide a. For example, in certain embodiments, the invention provides methods for producing therapeutic compositions having full-length mRNA encoding antigens from infectious agents such as viruses. In certain embodiments, the invention provides methods for producing therapeutic compositions having full-length mRNA encoding antigens from influenza virus. In certain embodiments, the invention provides methods for producing therapeutic compositions having full-length mRNA encoding antigens from respiratory syncytial virus. In certain embodiments, the invention provides methods for producing therapeutic compositions having full-length mRNA encoding antigens from rabies virus. In certain embodiments, the invention provides methods for producing therapeutic compositions having full-length mRNA encoding antigens from cytomegalovirus. In certain embodiments, the present invention provides methods for producing therapeutic compositions having full-length mRNA encoding antigens from rotavirus. In certain embodiments, the present invention provides methods for producing therapeutic compositions having full-length mRNA encoding antigens from hepatitis viruses, such as hepatitis A virus, hepatitis B virus, or hepatitis C virus. offer. In certain embodiments, the invention provides methods for producing therapeutic compositions having full-length mRNA encoding antigens from human papillomavirus. In certain embodiments, the invention provides methods for producing therapeutic compositions having full-length mRNA encoding an antigen from a herpes simplex virus, such as herpes simplex virus type 1 or herpes simplex virus type 2. In certain embodiments, the present invention provides methods for producing therapeutic compositions having full-length mRNA encoding antigens from a human immunodeficiency virus, such as human immunodeficiency virus type 1 or human immunodeficiency virus type 2. offer. In certain embodiments, the invention provides methods for producing therapeutic compositions having full-length mRNA encoding antigens from human metapneumovirus. In certain embodiments, the invention provides therapeutic compositions having full-length mRNA encoding an antigen from a human parainfluenza virus, such as human parainfluenza virus type 1, human parainfluenza virus type 2, or human parainfluenza virus type 3. A method for producing an object is provided. In certain embodiments, the invention provides methods for producing therapeutic compositions having full-length mRNA encoding antigens from malaria viruses. In certain embodiments, the present invention provides methods for producing therapeutic compositions having full-length mRNA encoding antigens from Zika virus. In certain embodiments, the present invention provides methods for producing therapeutic compositions having full-length mRNA encoding antigens from Chikungunyavirus.

In certain embodiments, the present invention provides methods for producing therapeutic compositions having full-length mRNA encoding antigens associated with cancer of a subject or identified from cancer cells of a subject. In certain embodiments, the present invention provides therapeutic compositions for making therapeutic compositions comprising full-length mRNAs encoding antigens determined from the subject's own cancer cells, i.e., for providing personalized cancer vaccines. provide a way. In certain embodiments, the invention provides methods for producing therapeutic compositions having full-length mRNA encoding antigens expressed from mutant KRAS genes.

In certain embodiments, the invention provides methods for producing therapeutic compositions having full-length mRNA encoding antibodies. In certain embodiments, an antibody can be a bispecific antibody. In certain embodiments, an antibody can be part of a fusion protein. In certain embodiments, the invention provides methods for producing therapeutic compositions having full-length mRNA encoding antibodies against OX40. In certain embodiments, the invention provides methods for producing therapeutic compositions having full-length mRNA encoding antibodies against VEGF. In certain embodiments, the invention provides methods for producing therapeutic compositions having full-length mRNA encoding antibodies to tissue necrosis factor-alpha. In certain embodiments, the invention provides methods for producing therapeutic compositions having full-length mRNA encoding antibodies to CD3. In certain embodiments, the invention provides methods for producing therapeutic compositions having full-length mRNA encoding antibodies against CD19.

In certain embodiments, the invention provides methods for producing therapeutic compositions having full-length mRNAs encoding immunomodulators. In certain embodiments, the invention provides methods for producing therapeutic compositions having full-length mRNA encoding interleukin-12. In certain embodiments, the invention provides methods for producing therapeutic compositions having full-length mRNA encoding interleukin-23. In certain embodiments, the invention provides methods for producing therapeutic compositions having full-length mRNA encoding interleukin-36 gamma. In certain embodiments, the present invention provides methods for producing therapeutic compositions having full-length mRNA encoding one or more constitutively active variants of interferon gene stimulating factor (STING) proteins. .

In certain embodiments, the invention provides methods for producing therapeutic compositions having full-length mRNA encoding an endonuclease. In certain embodiments, the invention provides methods for producing therapeutic compositions having full-length mRNAs encoding RNA guide DNA endonuclease proteins, such as Cas 9 proteins. In certain embodiments, the invention provides methods for producing therapeutic compositions having full-length mRNA encoding meganuclease proteins. In certain embodiments, the invention provides methods for producing therapeutic compositions having full-length mRNA encoding transcription activator-like effector nuclease proteins. In certain embodiments, the invention provides methods for producing therapeutic compositions having full-length mRNA encoding zinc finger nuclease proteins.

Methods of Delivery The delivery routes used in the methods of the invention allow non-invasive, self-administration of the compounds of the invention. In some embodiments, the method comprises intratracheal administration or intrapulmonary administration by aerosolization, nebulization, or instillation of a composition comprising mRNA encoding a therapeutic protein in a suitable transfection or lipid carrier vehicle as described above. with dosing. In some embodiments, proteins are encapsulated in liposomes. In some embodiments, the liposome comprises a lipid that is a compound of the invention. As used hereinafter, administration of a compound of the invention includes administration of a composition comprising the compound of the invention.

Although the local cells and tissues of the lung represent potential targets that can serve as biological depots or reservoirs for the production and secretion of proteins encoded by mRNA, Applicants believe that the compounds of the present invention We have found that pulmonary administration via aerosolization, nebulization, or instillation distributes even non-secreted proteins to the outside of lung cells. While not intending to be bound by any particular theory, the nanoparticle compositions of the present invention cross the lung-airway-blood barrier and penetrate intact nanoparticles into, e.g., heart, liver, spleen, and the like. It is contemplated to effect translation into non-lung cells and tissues, resulting in production of the encoded protein in these non-lung tissues. Thus, the utility of the compounds of the invention and methods of the invention extends beyond the production of therapeutic proteins in lung cells and tissue, and can be used for delivery to non-pulmonary target cells and/or tissues. They are useful in the management and treatment of numerous diseases, particularly peripheral diseases (eg, one or more lysosomal storage disorders) resulting from both secreted and non-secreted protein and/or enzyme deficiencies. In certain embodiments, the compounds of the invention used in the methods of the invention affect the distribution of mRNA-encapsulated nanoparticles and the production of the encoded proteins in liver, spleen, heart, and/or other non-lung cells. Bring. Pulmonary administration of a compound of the invention, for example, by aerosolization, nebulization, or instillation, results in the composition itself and its protein products (e.g., functional beta-galactosidase protein) affecting both local cells and tissues of the lung, and Translocation of the mRNA and delivery vehicle to non-lung cells results in detectability in peripheral target cells, tissues and organs.

In certain embodiments, compounds of the invention may be used in methods of the invention to specifically target peripheral cells or tissues. It is contemplated that following pulmonary delivery, the compounds of the invention will cross the pulmonary airway-blood barrier and distribute to cells other than local lung cells. Accordingly, the compounds disclosed herein can be administered to a subject by the pulmonary route of administration using a variety of approaches known to those of skill in the art (e.g., inhalation) to target local target cells and tissues of the lung, as well as peripheral non-target cells. It can be distributed in both lung cells and tissues such as liver, spleen, kidney, heart, skeletal muscle, lymph node, brain, cerebrospinal fluid, and plasma cells. As a result, both local and peripheral non-lung cells of the lung can function as biological reservoirs or depots capable of producing and/or secreting translation products encoded by one or more polynucleotides. can. Thus, the present invention is not limited to the treatment of pulmonary diseases or conditions, but rather delivery of polynucleotides to peripheral organs, tissues and cells (e.g., hepatocytes) that would otherwise only be achieved by systemic administration, or It can be used as a non-invasive means of promoting production of the enzymes and proteins encoded thereby. Exemplary peripheral non-lung cells include, but are not limited to, hepatocytes, epithelial cells, hematopoietic cells, epithelial cells, endothelial cells, osteocytes, stem cells, mesenchymal cells, nerve cells, cardiac cells, adipocytes, blood vessels Smooth muscle cells, cardiomyocytes, skeletal muscle cells, beta cells, pituitary cells, synovial lining cells, ovarian cells, testicular cells, fibroblasts, B cells, T cells, reticulocytes, leukocytes, granulocytes, and tumor cells is mentioned.

After administration of the composition to the subject, the protein product (e.g., functional protein or enzyme) encoded by the mRNA is detectable in peripheral target tissues for at least about 1-7 days or more after administration of the compound to the subject. is. The amount of protein product required to achieve therapeutic effect will vary depending on the condition being treated, the protein encoded, and the condition of the patient. For example, the protein product is at least about 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, at least 0.025 to 1.5 μg/ml (e.g., at least 0.050 μg/ml, at least 0.075 μg/ml, at least 0.1 μg/ml, at least 0.2 μg/ml, at least 0.3 μg/ml, at least 0.4 μg/ml, at least 0.5 μg/ml, at least 0 .6 μg/ml, at least 0.7 μg/ml, at least 0.8 μg/ml, at least 0.9 μg/ml, at least 1.0 μg/ml, at least 1.1 μg/ml, at least 1.2 μg/ml, at least 1.0 μg/ml. Concentrations (eg, therapeutic concentrations) of 3 μg/ml, at least 1.4 μg/ml, or at least 1.5 μg/ml) may be detectable in peripheral target tissues.

Nucleic acids can be administered via intratracheal administration of a liquid suspension of the compound and inhalation of an aerosol mist produced by a liquid nebulizer, or as described in US Pat. No. 5,780,014, incorporated herein by reference. It has been demonstrated that it can be delivered to the lung through the use of a dry powder device.

In certain embodiments, the compounds of the invention may be formulated so that they can be aerosolized or otherwise delivered as particulate liquids or solids prior to or at the time of administration to a subject. Such compounds are one for administering such solid or liquid particle compositions (e.g., aerosolized aqueous solutions or suspensions, etc.) to produce particles that are readily respirable or inhalable by a subject. or may be administered with the aid of any number of suitable devices. In some embodiments, such devices (e.g., metered dose inhalers, jet nebulizers, ultrasonic nebulizers, dry powder inhalers, propellant-based inhalers or injectors) deliver a predetermined mass of composition to a subject, It facilitates administration of volumes, or doses (eg, about 0.5 mg/kg of mRNA per dose). For example, in certain embodiments, a compound of the invention is administered to a subject using a metered dose inhaler containing a suspension or solution containing the compound and a suitable propellant. In certain embodiments, the compounds of the invention may be formulated as particulate powders (eg, respirable dry particles) intended for inhalation. In certain embodiments, compositions of the invention formulated as respirable particles are appropriately sized so that they may be respirable by a subject or delivered using a suitable device. (e.g., particles with an average D50 or D90 less than or equal to about 500 μm, 400 μm, 300 μm, 250 μm, 200 μm, 150 μm, 100 μm, 75 μm, 50 μm, 25 μm, 20 μm, 15 μm, 12.5 μm, 10 μm, 5 μm, 2.5 μm) size). In still other embodiments, the compounds of the invention are formulated to include one or more lung surfactants (eg, lamellar bodies). In some embodiments, the compounds of the invention are at least 0.05 mg/kg, at least 0.1 mg/kg, at least 0.5 mg/kg, at least 1.0 mg/kg, at least 2.0 mg/kg, at least 3 .0 mg/kg, at least 4.0 mg/kg, at least 5.0 mg/kg, at least 6.0 mg/kg, at least 7.0 mg/kg, at least 8.0 mg/kg, at least 9.0 mg/kg, at least 10 mg/kg at least 15 mg/kg, at least 20 mg/kg, at least 25 mg/kg, at least 30 mg/kg, at least 35 mg/kg, at least 40 mg/kg, at least 45 mg/kg, at least 50 mg/kg, at least 55 mg/kg, at least 60 mg/kg kg, at least 65 mg/kg, at least 70 mg/kg, at least 75 mg/kg, at least 80 mg/kg, at least 85 mg/kg, at least 90 mg/kg, at least 95 mg/kg, or at least 100 mg/kg body weight at a single dose administered to the subject as administered in In some embodiments, the compound of the invention is at least 0.1 mg, at least 0.5 mg, at least 1.0 mg, at least 2.0 mg, at least 3.0 mg, at least 4.0 mg, at least 5.0 mg, at least 6 .0 mg, at least 7.0 mg, at least 8.0 mg, at least 9.0 mg, at least 10 mg, at least 15 mg, at least 20 mg, at least 25 mg, at least 30 mg, at least 35 mg, at least 40 mg, at least 45 mg, at least 50 mg, at least 55 mg, at least 60 mg , at least 65 mg, at least 70 mg, at least 75 mg, at least 80 mg, at least 85 mg, at least 90 mg, at least 95 mg, or at least 100 mg of mRNA is administered to the subject such that it is administered in single or multiple doses.

Although certain compounds, compositions, and methods of this invention have been specifically described according to certain embodiments, the following examples serve only to illustrate and demonstrate the compounds of this invention. It is not intended to be limiting.

３－（ジメチルアミノ）プロピル４－ヒドロキシ－３－メトキシ安息香酸塩の合成（３）

Synthetic scheme of vanillic acid lipids

Synthesis of 3-(dimethylamino)propyl 4-hydroxy-3-methoxybenzoate (3)

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

Synthesis of 3-(dimethylamino)propyl 4-((4-(bis(2-((tert-butyldimethylsilyl)oxy)dodecyl)amino)butanoyl)oxy)-3-methoxybenzoate (4)

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

Synthesis of 3-(dimethylamino)propyl 4-((4-(bis(2-hydroxydodecyl)amino)butanoyl)oxy)-3-methoxybenzoate (VA-3-E12-DMAPr)

3-(dimethylamino)propyl 4-((4-((bis(2-((tert-butyldimethylsilyl)oxy)dodecyl)amino)butanoyl)oxy)-3-methoxybenzoate 4 in 10 mL tetrahydrofuran To a solution of (330 mg, 0.35 mmol) was added dropwise hydrofluoric acid (70%, 2.5 mL) in pyridine at 0° C. and the reaction mixture was stirred overnight at room temperature.Saturated sodium bicarbonate solution. was added to pH=7-8 and the mixture was extracted with ethyl acetate.The organic layer was washed with brine and dried over anhydrous sodium sulfate.After filtration and concentration, the crude product was subjected to reverse phase column chromatography ( C18: 5-95% MeCN/water/0.1% TFA) to give 3-(dimethylamino)propyl 4-((4-(bis(2-hydroxydodecyl)amino)butanoyl)oxy)-3- Methoxybenzoate was obtained as the TFA salt (120 mg, 48%) and stored in 2-butanol to prevent decomposition.

All other lipids were prepared in similar yields following representative procedures.

３－（ジメチルアミノ）プロピル４－ヒドロキシ－３，５－ジメトキシ安息香酸塩の合成（６）

Synthetic scheme of syringate lipids

Synthesis of 3-(dimethylamino)propyl 4-hydroxy-3,5-dimethoxybenzoate (6)

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

Synthesis of 3-(dimethylamino)propyl 4-((4-(bis(2-((tert-butyldimethylsilyl)oxy)dodecyl)amino)butanoyl)oxy)-3,5-dimethoxybenzoate (7)

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

Synthesis of 3-(dimethylamino)propyl 4-((4-(bis(2-hydroxydodecyl)amino)butanoyl)oxy)-3,5-dimethoxybenzoate (SA-3-E12-DMAPr)

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

All other lipids were prepared in similar yields following representative procedures.

３－（ジメチルアミノ）プロピル（Ｅ）－３－（４－ヒドロキシ－３，５－ジメトキシフェニル）アクリレートの合成（９）

Synthetic scheme of sinapinic acid lipid

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

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

(E)-4-(3-(3-(dimethylamino)propoxy)-3-oxoprop-1-en-1-yl)-2,6-dimethoxyphenyl 4-(bis(2-((tert-butyl) Synthesis of dimethylsilyl)oxy)dodecyl)amino)butanoate (10)

To a solution of 4-(bis(2-((tert-butyldimethylsilyl)oxy)dodecyl)amino)butanoic acid AIM-3-E12 (1.8 g, 2.59 mmol) in 20 mL of dichloromethane was added chloride at 0 °C. Oxalyl (0.3 mL, 3.09 mmol) was added followed by dimethylformamide (1 drop) and the mixture was stirred at 0° C. for 2 hours. The reaction mixture was evaporated to dryness and the residue dissolved in 20 mL of dichloromethane. After cooling to 0° C., 3-(dimethylamino)propyl (E)-3-(4-hydroxy-3,5-dimethoxyphenyl)acrylate 9 (0.4 g, 1.29 mmol) was added followed by pyridine (0.29 mmol). 62 mL, 7.7 mmol) was added and the reaction mixture was stirred overnight at room temperature. Ice was added to quench the reaction and the organic layer was washed with brine and dried over anhydrous sodium sulfate. After filtration and concentration, the crude is purified by flash chromatography to give (E)-4-(3-(3-(dimethylamino)propoxy)-3-oxoprop-1-en-1-yl)-2, 6-Dimethoxyphenyl 4-(bis(2-((tert-butyldimethylsilyl)oxy)dodecyl)amino)butanoate 10 was obtained as a pale yellow oil (540 mg, 42%).

(E)-4-(3-(3-(dimethylamino)propoxy)-3-oxoprop-1-en-1-yl)-2,6-dimethoxyphenyl 4-(bis(2-hydroxydodecyl)amino) Synthesis of butanoate (SI-3-E12-DMAPr)

(E)-4-(3-(3-(dimethylamino)propoxy)-3-oxoprop-1-en-1-yl)-2,6-dimethoxyphenyl 4-(bis(2- To a solution of ((tert-butyldimethylsilyl)oxy)dodecyl)amino)butanoate 10 (540 mg, 0.55 mmol) at 0° C. was added hydrofluoric acid (70%, 2.5 mL) in pyridine dropwise. , the reaction mixture was stirred overnight at room temperature. Saturated sodium bicarbonate solution was added to pH=7-8 and the mixture was extracted with ethyl acetate. The organic layer was washed with brine and dried over anhydrous sodium sulfate. After filtration and concentration, the crude product was purified by reverse-phase column chromatography (C18: 5-95% MeCN/water/0.1% TFA) to give (E)-4-(3-(3-( Dimethylamino)propoxy)-3-oxoprop-1-en-1-yl)-2,6-dimethoxyphenyl 4-(bis(2-hydroxydodecyl)amino)butanoate was obtained as a TFA salt (330 mg, 80%) .

All other lipids were prepared in similar yields following representative procedures.

合成スキームＡにおける中間体（３ａ）の合成：

Synthetic scheme A of phenolic acid lipid

Synthesis of Intermediate (3a) in Synthetic Scheme A:

To a solution of (1) (2.00 g, 2.86 mmol) in anhydrous CH ₂ Cl ₂ (10 mL) at 0° C. was added dropwise oxalyl chloride (0.98 mL, 4.0 eq) to turn the reaction mixture into It was gradually warmed to room temperature and stirred for 2 hours. After removing excess solvent and oxalyl chloride under reduced pressure, the remaining residue was redissolved in anhydrous CH ₂ Cl ₂ (30 mL). To the stirred acid chloride solution at 0° C. was added (2a) (555 mg, 1.0 eq), DMAP (349 mg, 1.0 eq), followed by triethylamine (3.18 mL, 8.0 eq). The reaction mixture was slowly warmed to room temperature and then stirred at the same temperature for 16 hours. After 16 hours, the reaction mixture was diluted with CH ₂ Cl ₂ and washed with saturated NaHCO _3(aq) solution. The separated organic layer was washed with brine, dried over _Na2SO4 and concentrated _under reduced pressure to deliver crude material. The crude material was first purified using ₅₀ % EtOAc in hexanes, then re-purified with 15% EtOAc in _CH2Cl2 to give (3a) (623 mg, 25%) as a viscous oil. rice field. MS (ESI+) calcd for _C50H93NO7Si2 , [M+H] ⁺ = _876.65 , found _{= 876.6} .

Synthesis of Intermediate (3b) in Synthetic Scheme A:

Using (2b) and following the procedure of (3a), (3b) (557 mg, 23%) was obtained as a viscous oil. MS (ESI+) calcd for _C49H91NO6Si2 , [M+H] ⁺ = _846.64 , found _{= 846.6} .

Synthesis of Intermediate (5a) in Synthetic Scheme A:

To a solution of (3a) (308 mg, 0.351 mmol) in anhydrous CH ₂ Cl ₂ (3 mL) was added oxalyl chloride (0.15 mL, 5.0 eq) at room temperature and stirred at the same temperature for 2 hours. After removing excess solvent and oxalyl chloride under reduced pressure, the remaining residue was redissolved in anhydrous CH ₂ Cl ₂ (3 mL). To the stirred acid chloride solution at 0° C. was added 3-dimethylaminopropanol (4) (109 mg, 3 eq) followed by triethylamine (0.10 mL, 2.0 eq). The reaction mixture was slowly warmed to room temperature and then stirred at the same temperature for 16 hours. After completion of the reaction monitored by MS, the reaction mixture was concentrated under reduced pressure to dryness and purified using 0-10% MeOH in CH ₂ Cl ₂ to give (5a) (204 mg, 60%). was obtained as a viscous oil. MS (ESI+) calcd _{for C55H104N2O7Si2} , [M+H] ⁺ _{= 961.74} , _found = 961.7.

Synthesis of Intermediate (5b) in Synthetic Scheme A:

Using (3b), following the procedure of (5a) gave (5b) (248 mg, 90%) as a viscous oil. MS ( _ESI +) calcd _{for C54H102N2O6Si2} , [M+H] ⁺ _{= 931.73} , found = 931.7.

Synthesis of TBL-0731 Compound 355 (6a) in Synthetic Scheme A:

To a stirred solution of (5a) (204 mg, 0.212 mmol) in anhydrous THF (3 mL) at 0° C. was added 70% hydrogen fluoride in pyridine (1.09 mL, 197 eq) dropwise and slowly warmed to room temperature. was warmed to The reaction mixture was stirred at room temperature for 16 hours. After completion of the reaction monitored by MS, the reaction mixture was cooled to 0° C. and quenched with batchwise addition of solid NaHCO ₃ . After gas formation was minimized, the resulting mixture was diluted with EtOAc and neutralized with saturated NaHCO _3(aq) solution. The separated organic layer was washed with brine, dried over _Na2SO4 and concentrated _under reduced pressure to deliver crude material. The crude material was purified using 0-20% MeOH in CH ₂ Cl ₂ to give TBL-0731 (6a) (132 mg, 85%) as a viscous oil. MS ( _ESI +) calcd for _C43H76N2O7 , [M+H] ^<+> _{= 733.57} , found = 733.5. ¹ H NMR (500 MHz, CDCl ₃ ) δ 7.64 (d, J = 15.9 Hz, 1 H), 7.14-7.08 (m, 2 H), 7.05 (d, J = 8.1, 3 .5Hz, 1H), 6.37 (d, J = 16.0Hz, 1H), 4.27 (t, J = 6.4Hz, 2H), 3.86 (s, 3H), 3.72-3 .63 (m, 2H), 2.78-2.70 (m, 2H), 2.67-2.47 (m, 6H), 2.46-2.40 (m, 2H), 2.36 (s, 6H), 2.02-1.90 (m, 4H), 1.49-1.35 (m, 4H), 1.34-1.15 (m, 32H), 0.87 (t , J=6.9 Hz, 6H).

Synthesis of TBL-0750 Compound 467 (6b) in Synthetic Scheme A:

Using (5b) and following the procedure of (6a), TBL-0750 (6b) (153 mg, 82%) was obtained as a viscous oil. MS ( _ESI +) calcd for _C42H74N2O6 , [M+H] ^<+> _{= 703.55} , found = 703.6. ¹ H NMR (400 MHz, CDCl ₃ ) δ 7.66 (d, J=16.0 Hz, 1 H), 7.53 (d, J=8.7, 1.3 Hz, 2 H), 7.12 (d, J = 8.6, 1.5Hz, 2H), 6.38 (d, J = 16.0, 0.9Hz, 1H), 4.26 (t, J = 6.4Hz, 2H), 3.70- 3.63 (m, 2H), 2.73-2.64 (m, 2H), 2.64-2.46 (m, 6H), 2.45-2.40 (m, 2H), 2. 34 (s, 6H), 1.99-1.88 (m, 4H), 1.43-1.34 (m, 4H), 1.31-1.21 (m, 32H), 0.87 ( t, J=6.8 Hz, 6H).

合成スキームＢにおける中間体（３）の合成：

Synthetic scheme B of phenolic acid lipid

Synthesis of Intermediate (3) in Synthetic Scheme B:

To a solution of acid intermediate (1) (4.58 g) in CH ₂ Cl ₂ (30 mL, anhydrous) was added oxalyl chloride (1.04 mL, 2 eq) and stirred at room temperature for 2 hours. All volatiles were removed under reduced pressure and the remaining residue was redissolved in CH ₂ Cl ₂ (30 mL, anhydrous). To this solution was then added syringic acid (2) (1.32 g, 1.1 eq) followed by pyridine (2.93 mL, 6 eq) and the resulting mixture was stirred overnight at room temperature. After stirring overnight, the solvent was removed under reduced pressure and the remaining residue was solubilized with minimal CH ₂ Cl ₂ and filtered through a short silica gel plug with 100% EtOAc as eluent. The clear filtrate was concentrated to dryness to give crude material, which was purified by MPLC using a 10-100% EtOAc in hexanes gradient over 10 CV to give the acid product (3) (3.70 g, 78%). Obtained. MS (ESI+ ₎ calcd for _{C53H101NO8Si2} , [M+H] ⁺ = _936.7 , _found = 936.7.

Synthesis of Intermediate (5a) in Synthetic Scheme B:

To a solution of benzoic acid (3) (400 mg) in CH ₂ Cl ₂ (4 mL, anhydrous) was added oxalyl chloride (0.18 mL, 5 eq) and stirred at room temperature for 2 hours. All volatiles were removed under reduced pressure and the remaining residue was redissolved in CH ₂ Cl ₂ (3 mL, anhydrous). The resulting solution was cooled in an ice bath and then a solution of 2-aminoethanol (4a) (76 mg) in CH ₂ Cl ₂ (1 mL) was added. The reaction mixture was warmed to room temperature and stirred at the same temperature for 16 hours. After complete consumption of the starting material, monitored by LC-MS, the reaction mixture was concentrated to dryness and the crude material was purified by MPLC using a 0-20% MeOH gradient in CH ₂ Cl ₂ over 12 CV to give the product (5a) was obtained as a viscous oil (225 mg, 52%). MS ( _ESI +) calcd _{for C57H110N2O8Si2} , [M+H] ⁺ _{= 1007.8} , found = 1007.6.

Synthesis of Intermediate (5b) in Synthetic Scheme B:

Following the synthetic procedure for (5a) using 4-aminobutanol (4b) gave the product (5b) as a viscous oil (360 mg, 81%). MS _{( ESI} +) calcd _{for C59H114N2O8Si2} , [M+H] ⁺ = _1035.8 , found = 1035.6.

Synthesis of Intermediate (5c) in Synthetic Scheme B:

Following the synthetic procedure of (5a) using intermediate (3) (500 mg) and 3-morpholinopropanol (4c) gave product (5c) (350 mg, 62%). MS _{( ESI +} ) calcd for _{C60H114N2O9Si2} , [M+H] ⁺ = _1063.8 , found = 1063.6.

Synthesis of Intermediate (5d) in Synthetic Scheme B:

Following the synthetic procedure of (5a) using intermediate (3) (500 mg) and 2-pyridinylethanol (4d) gave product (5d) (237 mg, 43%). MS _{( ESI} +) calcd _{for C60H108N2O8Si2} , [M+H] ⁺ = _1041.8 , found = 1041.6.

Synthesis of Intermediate (5e) in Synthetic Scheme B:

Following the procedure for synthesis of 5a using intermediate (3) (843 mg) and 4-methylpiperazinylethanol (4e) gave product (5e) (346 mg, 36%). MS (ESI+) calcd for _{C60H115N3O8Si2 ,} [M+H] ⁺ _{= 1062.8} , _found = 1062.7.

Synthesis of TBL-0507 Compound 48 (6a) in Synthetic Scheme B:

To a stirred solution of TBS-protected intermediate (5a) (225 mg) in THF (3 mL, anhydrous) in a plastic polymer scintillation vial (non-glass) was added triethylamine (0.16 mL, 5 eq) followed by triethylamine-3HF. (0.36 mL, 10 eq) was added dropwise. The reaction mixture was stirred at 50° C. overnight. The reaction was monitored by suspending two drops of the reaction mixture between layers of EtOAc and NaHCO _3(aq) and analyzing the organic layer (TLC or LC-MS). Once the starting material was consumed, excess HF and volatiles were removed by blowing off with _N2 gas in a fume hood and the remaining material was diluted with EtOAc and neutralized with saturated aqueous _NaHCO3 (pH paper confirmation). The separated organic layer was washed with brine, dried over _Na2SO4 and concentrated _under reduced pressure to give crude material. The crude material was purified by MPLC using a 0-40% MeOH gradient in CH ₂ Cl ₂ over 10 CV to afford TBL-0507 (6a) (90 mg, 52%). MS ( _ESI +) calcd for _C45H82N2O8 , [M+H] ^<+ _{> =} 779.6, found = 779.5.

Synthesis of TBL-0508 Compound 49 (6b) in Synthetic Scheme B:

Using TBS protected intermediate (5b) (360 mg), following the procedure of (6a) gave TBL-0508 (6b) (71 mg, 25%). MS ( _ESI +) calcd for _C47H86N2O8 , [M+H] ⁺ = _{807.6 ,} found = 807.5.

Synthesis of TBL-0517 Compound 562 (6c) in Synthetic Scheme B:

TBS protected intermediate (5c) (350 mg) was used to purify TBL-0517 (6c) (164 mg, 60%) with a 0-10% MeOH gradient in CH ₂ Cl ₂ following the procedure in (6a). ). MS ₍ ESI+) calcd for _C48H86N2O9 , [M+H] ⁺ = _835.6 , found ₌ 835.5.

Synthesis of TBL-0518 Compound 563 (6d) in Synthetic Scheme B:

The TBS protected intermediate (5d) (237 mg) was used to purify TBL-0518 (6d) (111 mg, 60%) with a 0-10% MeOH gradient in CH ₂ Cl ₂ following the procedure in (6a). ). MS ( _ESI +) calcd for _C48H80N2O8 , [M+H] ⁺ = _{813.6 ,} found = 813.5.

Synthesis of TBL-0535 Compound 564 (6e) in Synthetic Scheme B:

The TBS protected intermediate (5e) (346 mg) was used to purify TBL-0535 (6e) (88 mg, 32%) with a 0-20% MeOH gradient in CH ₂ Cl ₂ following the procedure in (6a). ). MS (ESI+) calcd for _C48H87N3O8 , [M+H] ⁺ _{= 834.6} , found = _834.6 .

合成スキームＣにおける中間体（９ａ）の合成：

Synthetic scheme C of phenolic acid lipid

Synthesis of Intermediate (9a) in Synthetic Scheme C:

Isopropanol (5 mL) and triethylamine (1.35 mL, 2 eq) were added to a flask containing amino acid (7) (500 mg) and dodecyl acrylate (8a) (2.91 g, 2.5 eq). The resulting mixture was heated at 90° C. for 3 hours. After completion of the reaction monitored by MS, the reaction mixture was cooled to room temperature and concentrated under reduced pressure. The remaining material was purified by MPLC using 0-12% MeOH in CH ₂ Cl ₂ to give product (9a) (987 mg, 35%). MS ( _{ESI+) calcd for C34H65NO6 ,} [M+H] ⁺ = 584.5, found = 584.5 _.

Synthesis of Intermediate (9b) in Synthetic Scheme C:

(9a) using amino acid (7) (1.00 g), tetradecyl acrylate (8b) (6.51 g, 2.5 eq), isopropanol (10 mL), and triethylamine (2.70 mL, 2 eq). to give the product (9b) (1.80 g, 29%). MS ( _ESI +) calcd for _C38H73NO6 , [M+H] ⁺ = 640.5 _, found = 640.5.

Synthesis of TBL-0484 Compound 565 (11a) in Synthetic Scheme C:

To a solution of intermediate (9a) (300 mg) in anhydrous CH ₂ Cl ₂ (3 mL) at room temperature was added oxalyl chloride (1.0 mL, 23 eq) and stirred at the same temperature for 2 hours. After removing excess solvent and oxalyl chloride under reduced pressure, the remaining residue was redissolved in anhydrous CH ₂ Cl ₂ (3 mL). Phenolic acid (10) (146 mg, 1.0 eq) and pyridine (0.21 mL, 5 eq) were added to the stirred acid chloride solution at room temperature and stirred at the same temperature for 16 hours. After completion of the reaction monitored by MS, the reaction mixture was concentrated under reduced pressure. The remaining crude material was purified by MPLC using 0-10% MeOH in CH ₂ Cl ₂ to give TBL-0484 (11a) (90 mg, 21%). MS ( _ESI +) calcd for _C48H84N2O10 , [M+H] ^<+> _{= 849.6} , found = 849.5.

Synthesis of TBL-0485 Compound 566 (11b) in Synthetic Scheme C:

Using intermediate (9b) (300 mg) and following the procedure of (11a), TBL-0485 (11b) (80 mg, 19%) was obtained. MS ( _ESI +) calcd for _C52H92N2O10 , [M+H] ⁺ = _905.7 , _found = 905.6.

Example 1 Lipid Nanoparticle Formulations The cationic lipids described herein can be used in the preparation of lipid nanoparticles according to methods known in the art. For example, suitable methods include those described in International Publication No. WO2018/089801, which is incorporated herein by reference in its entirety.

One exemplary process for formulation of lipid nanoparticles is Process A of WO2018/089801 (see, eg, Example 1 and Figure 1 of WO2018/089801). Process A (“A”) relates to the conventional method of encapsulating mRNA by mixing it with a lipid mixture without first preforming the lipid into lipid nanoparticles. In an exemplary process, an ethanol lipid solution and an aqueous mRNA buffer were prepared separately. Solutions of lipid mixtures (cationic lipids, helper lipids, zwitterionic lipids, PEG lipids, etc.) were prepared by dissolving the lipids in ethanol. An mRNA solution was prepared by dissolving mRNA in citrate buffer. Both mixtures were then heated to 65° C. and then mixed. These two solutions were then mixed using a pump system. In some cases, these two solutions were mixed using a gear pump system. In certain embodiments, these two solutions were mixed using a "T" joint (or "Y" joint). The mixture was then purified by diafiltration using the TFF process. The resulting formulation was concentrated and stored at 2-8°C until further use.

A second exemplary process for formulation of lipid nanoparticles is Process B of WO2018/089801 (see, eg, Example 2 and Figure 2 of WO2018/089801). Process B (“B”) relates to the process of encapsulating messenger RNA (mRNA) by mixing preformed lipid nanoparticles with mRNA. A different range of conditions can be used in Process B, eg, different temperatures (ie, the mixture is heated or not heated), buffers, and concentrations. In an exemplary process, lipids dissolved in ethanol and citrate buffer were mixed using a pump system. Instantaneous mixing of these two streams resulted in the formation of hollow lipid nanoparticles, which was a self-assembly process. The resulting formulation mixture was hollow lipid nanoparticles in citrate buffer containing alcohol. The formulation was then subjected to a TFF purification process in which buffer exchange was performed. The resulting preformed empty lipid nanoparticle suspension was then mixed with mRNA using a pump system. For certain cationic lipids, heating the solution after mixing increased the percentage of lipid nanoparticles containing mRNA and increased the overall yield of mRNA.

The lipid nanoparticle formulations in Table 5 were prepared by either Process A or B. Each formulation contained the mRNA encoding firefly luciferase protein (FFL mRNA), and lipids (cationic lipid: DMG-PEG2000, cholesterol: DOPE), as described in Table 5 (mole % ratio).

Delivery of FFL mRNA by intratracheal administration. (age) under anesthesia. Approximately 24 hours after dosing, animals were dosed with luciferin at 150 mg/kg (60 mg/mL) by intraperitoneal injection at 2.5 mL/kg. After 5-15 minutes, all animals were imaged using the IVIS imaging system to measure luciferase production in the lungs. Figure 1 shows that lipid nanoparticles containing cationic lipids described herein are effective in delivering FFL mRNA in vivo based on positive luciferase activity.

式中、Ｌ_１は結合、（Ｃ_１－Ｃ_６）アルキルまたは（Ｃ_２－Ｃ_６）アルケニルであり、
式中、ＸはＯまたはＳであり、
式中、Ｒ^１、Ｒ^２、Ｒ^３、Ｒ^４およびＲ^５は、それぞれ独立してＨ、ＯＨ、任意で置換された（Ｃ_１－Ｃ_６）アルキル、任意で置換された（Ｃ_２－Ｃ_６）アルケニル、任意で置換された（Ｃ_２－Ｃ_６）アルキニル、任意で置換された（Ｃ_１－Ｃ_６）アルコキシおよび－ＯＣ（Ｏ）Ｒ’から選択され、
式中、Ｒ^１、Ｒ^２、Ｒ^３、Ｒ^４またはＲ^５の少なくとも一つは、－ＯＣ（Ｏ）Ｒ’であり、
式中、Ｒ’は以下であり、

式中、Ｒ^６は以下であり、

式中、ｍおよびｐはそれぞれ独立して、０、１、２、３、４、または５であり、
式中、Ｒ^７は、Ｈ、任意で置換された（Ｃ_１－Ｃ_６）アルキル、任意で置換された（Ｃ_２－Ｃ_６）アルケニル、任意で置換された（Ｃ_２－Ｃ_６）アルキニル、任意で置換された（Ｃ_１－Ｃ_６）アシル、－（ＣＨ_２）_ｋＲ^Ａまたは－（ＣＨ_２）_ｋＣＨ（ＯＲ^１１）Ｒ^Ａから選択され、
式中、Ｒ^８は、Ｈ、任意で置換された（Ｃ_１－Ｃ_６）アルキル、任意で置換された（Ｃ_２－Ｃ_６）アルケニル、任意で置換された（Ｃ_２－Ｃ_６）アルキニル、任意で置換された（Ｃ_１－Ｃ_６）アシル、－（ＣＨ_２）_ｎＲ^Ｂまたは－（ＣＨ_２）_ｎＣＨ（ＯＲ^１２）Ｒ^Ｂから選択され、
式中、Ｒ^９は、Ｈ、任意で置換された（Ｃ_１－Ｃ_６）アルキル、任意で置換された（Ｃ_２－Ｃ_６）アルケニル、任意で置換された（Ｃ_２－Ｃ_６）アルキニル、任意で置換された（Ｃ_１－Ｃ_６）アシル、－（ＣＨ_２）_ｑＲ^Ｃまたは－（ＣＨ_２）_ｑＣＨ（ＯＲ^１３）Ｒ^Ｃから選択され、
式中、Ｒ^１０は、Ｈ、任意で置換された（Ｃ_１－Ｃ_６）アルキル、任意で置換された（Ｃ_２－Ｃ_６）アルケニル、任意で置換された（Ｃ_２－Ｃ_６）アルキニル、任意で置換された（Ｃ_１－Ｃ_６）アシル、－（ＣＨ_２）_ｒＲ^Ｄまたは－（ＣＨ_２）_ｒＣＨ（ＯＲ^１４）Ｒ^Ｄから選択され、
式中、ｋ、ｎ、ｑおよびｒはそれぞれ独立して１、２、３、４または５であり、
または式中、（ｉ）Ｒ^７およびＲ^８、または（ｉｉ）Ｒ^９およびＲ^１０は共に、任意で置換された５員または６員ヘテロシクロアルキルまたはヘテロアリールを形成し、ヘテロシクロアルキルまたはヘテロアリールは、Ｎ、ＯおよびＳから選択される１～３個のヘテロ原子を含み、
式中、Ｒ^１１、Ｒ^１２、Ｒ^１３、およびＲ^１４はそれぞれ独立してＨ、メチル、エチル、またはプロピルから選択され、
式中、Ｒ^Ａ、Ｒ^Ｂ、Ｒ^ＣおよびＲ^Ｄはそれぞれ独立して、任意で置換された（Ｃ_６－Ｃ_２０）アルキル、任意で置換された（Ｃ_６－Ｃ_２０）アルケニル、任意で置換された（Ｃ_６－Ｃ_２０）アルキニル、任意で置換された（Ｃ_６－Ｃ_２０）アシル、任意で置換された－ＯＣ（Ｏ）アルキル、任意で置換された－ＯＣ（Ｏ）アルケニル、任意で置換された（Ｃ_１－Ｃ_６）モノアルキルアミノ、任意で置換された（Ｃ_１－Ｃ_６）ジアルキルアミノ、任意で置換された（Ｃ_１－Ｃ_６）アルコキシ、－ＯＨ、－ＮＨ_２から選択され、
式中、Ｒ^７、Ｒ^８、Ｒ^９、Ｒ^１０のうちの少なくとも一つは、それぞれＲ^Ａ、Ｒ^Ｂ、Ｒ^ＣまたはＲ^Ｄ部分を含み、Ｒ^Ａ、Ｒ^Ｂ、Ｒ^ＣまたはＲ^Ｄは独立して、任意で置換された（Ｃ_６－Ｃ_２０）アルキル、任意で置換された（Ｃ_６－Ｃ_２０）アルケニル、任意で置換された（Ｃ_６－Ｃ_２０）アルキニル、任意で置換された（Ｃ_６－Ｃ_２０）アシル、任意で置換された－ＯＣ（Ｏ）（Ｃ_６－Ｃ_２０）アルキルまたは任意で置換された－ＯＣ（Ｏ）（Ｃ_６－Ｃ_２０）アルケニルから選択されるカチオン性脂質、
またはその薬学的に許容可能な塩。
２．任意のアルキル、アルケニル、アルキニル、アシル、アルコキシ、モノアルキルアミノ、ジアルキルアミノ、ヘテロシクロアルキルまたはヘテロアリールは、（Ｃ_１－Ｃ_６）アルキル、（Ｃ_２－Ｃ_６）アルケニル、（Ｃ_２－Ｃ_６）アルキニル、（Ｃ_１－Ｃ_６）アシル、（Ｃ_１－Ｃ_６）アルコキシ、ハロゲン、－ＣＯＲ、－ＣＯ_２Ｈ、－ＣＯ_２Ｒ、－ＣＮ、－ＯＨ、－ＯＲ、－ＯＣＯＲ、－ＯＣＯ_２Ｒ、－ＮＨ_２、－ＮＨＲ、－Ｎ（Ｒ）_２、－ＳＲまたは－ＳＯ_２Ｒからなる群から選択される一つまたは複数の置換基で任意に置換され、または炭素原子上の二つのジェミナル水素が、基＝ＮＨで置換され、Ｒの各例は独立してＣ_１－Ｃ_１０脂肪族アルキルである、番号付けされた実施形態１に記載のカチオン性脂質またはその薬学的に許容可能な塩。
３．
ｉ）Ｒ^ＡおよびＲ^Ｂは同一であり、および／または
ｉｉ）Ｒ^ＣおよびＲ^Ｄは同一である、番号付けされた実施形態１～２のいずれかに記載のカチオン性脂質またはその薬学的に許容可能な塩。
４．
ｉ）Ｒ^ＡおよびＲ^Ｂは異なり、および／または
ｉｉ）Ｒ^ＣおよびＲ^Ｄは異なる、番号付けされた実施形態１または２に記載のカチオン性脂質またはその薬学的に許容可能な塩。
５．Ｒ^Ａ、Ｒ^Ｂ、Ｒ^ＣおよびＲ^Ｄが同一である、番号付けされた実施形態１～３のいずれか一つに記載のカチオン性脂質またはその薬学的に許容可能な塩。
６．Ｒ^Ａ、Ｒ^Ｂ、Ｒ^ＣおよびＲ^Ｄのうちの一つまたは複数が異なる、番号付けされた実施形態１～４のいずれか一つに記載のカチオン性脂質またはその薬学的に許容可能な塩。
７．ＸがＯである、番号付けされた実施形態１～６のいずか一つに記載のカチオン性脂質またはその薬学的に許容可能な塩。
８．ＸがＳである、番号付けされた実施形態１～６のいずか一つに記載のカチオン性脂質またはその薬学的に許容可能な塩。
９．Ｒ^１、Ｒ^２、Ｒ^３、Ｒ^４およびＲ^５のうちの一つのみが－ＯＣ（Ｏ）Ｒ’である、番号付けされた実施形態１～８のいずれか一つに記載のカチオン性脂質またはその薬学的に許容可能な塩。
１０．Ｒ^１、Ｒ^２、Ｒ^３、Ｒ^４またはＲ^５のいずれもＯＨではない、番号付けされた実施形態９に記載のカチオン性脂質またはその薬学的に許容可能な塩。
１１．Ｒ^１、Ｒ^２、Ｒ^３、Ｒ^４およびＲ^５のうちの二つが－ＯＣ（Ｏ）Ｒ’である、番号付けされた実施形態１～８のいずれか一つに記載のカチオン性脂質またはその薬学的に許容可能な塩。
１２．Ｒ^１、Ｒ^２、Ｒ^３、Ｒ^４またはＲ^５のいずれもＯＨではない、番号付けされた実施形態１１に記載のカチオン性脂質またはその薬学的に許容可能な塩。
１３．Ｒ^１、Ｒ^２、Ｒ^３、Ｒ^４およびＲ^５のうちの三つが－ＯＣ（Ｏ）Ｒ’である、番号付けされた実施形態１～８のいずれか一つに記載のカチオン性脂質またはその薬学的に許容可能な塩。
１４．Ｒ^１および／またはＲ^５が－ＯＣ（Ｏ）Ｒ’である、番号付けされた実施形態１～１３のいずれか一つに記載のカチオン性脂質またはその薬学的に許容可能な塩。
１５．Ｒ^２および／またはＲ^４が－ＯＣ（Ｏ）Ｒ’である、番号付けされた実施形態１～１４のいずれか一つに記載のカチオン性脂質またはその薬学的に許容可能な塩。
１６．Ｒ^３が－ＯＣ（Ｏ）Ｒ’である、番号付けされた実施形態１～１５のいずれか一つに記載のカチオン性脂質またはその薬学的に許容可能な塩。
１７．
ｉ）ｐ、ｑ、およびｒは同一であり、または
ｉｉ）ｐ、ｑ、およびｒのうちの一つまたは複数が異なるか、または
ｉｉｉ）ｑおよびｒは同一で、ｐは異なる、番号付けされた実施形態１～１６のいずれか一つに記載のカチオン性脂質またはその薬学的に許容可能な塩。
１８．
ｉ）ｋ、ｍ、およびｎは同一であり、または
ｉｉ）ｋ、ｍおよびｎのうちの一つまたは複数が異なるか、または
ｉｉｉ）ｋおよびｎは同一であり、ｍは異なる、番号付けされた実施形態１～１７のいずれか一つに記載のカチオン性脂質またはその薬学的に許容可能な塩。
１９．ｍが１、２、または３である、番号付けされた実施形態１～１８のいずか一つに記載のカチオン性脂質またはその薬学的に許容可能な塩。
２０．ｐが１、２、または３である、番号付けされた実施形態１～１９のいずか一つに記載のカチオン性脂質またはその薬学的に許容可能な塩。
２１．Ｒ^’が以下である、番号付けされた実施形態１～２０のいずか一つに記載のカチオン性脂質またはその薬学的に許容可能な塩。

２２．
ｉ）ｋ、ｍおよびｎ＝１、または
ｉｉ）ｋ、ｍおよびｎ＝１、ならびにＲ^１１およびＲ^１２＝Ｈ、または
ｉｉｉ）ｋおよびｎ＝１、ならびにｍ＝２、または
ｉｖ）ｋおよびｎ＝１、ｍ＝２、ならびにＲ^１１およびＲ^１２＝Ｈ、または
ｖ）ｋおよびｎ＝１、ならびにｍ＝３、または
ｖｉ）ｋおよびｎ＝１、ｍ＝３、ならびにＲ^１１およびＲ^１２＝Ｈである、番号付けされた実施形態２１に記載のカチオン性脂質またはその薬学的に許容可能な塩。
２３．Ｒ^６が以下である、番号付けされた実施形態１～２２のいずか一つに記載のカチオン性脂質またはその薬学的に許容可能な塩。

２４．
ｉ）ｐ、ｑおよびｒ＝１、または
ｉｉ）ｐ、ｑおよびｒ＝１、ならびにＲ^１３およびＲ^１４はＨであり、または
ｉｉｉ）ｑおよびｒ＝１、およびｐ＝２、または
ｉｖ）ｑおよびｒ＝１、ｐ＝２、ならびにＲ^１３およびＲ^１４はＨである、番号付けされた実施形態２３に記載のカチオン性脂質またはその薬学的に許容可能な塩。
２５．Ｒ^６が以下からなる群から選択される、番号付けされた実施形態１～２２のいずか一つに記載のカチオン性脂質またはその薬学的に許容可能な塩。

２６．Ｒ^６が以下である、番号付けされた実施形態２５に記載のカチオン性脂質またはその薬学的に許容可能な塩。

２７．式（ＩＩ）に従う構造を有する番号付けされた実施形態１～２６のいずれか一つに記載のカチオン性脂質：

またはその薬学的に許容可能な塩。
２８．式（ＩＩＡ）に従う構造を有する番号付けされた実施形態２７に記載のカチオン性脂質

またはその薬学的に許容可能な塩。
２９．式（ＩＩＢ）、（ＩＩＣ）、（ＩＩＤ）、または（ＩＩＥ）のうちの一つに従う構造を有する、番号付けされた実施形態２８に記載のカチオン性脂質

またはその薬学的に許容可能な塩。
３０．式（ＩＩＦ）に従う構造を有する番号付けされた実施形態２７に記載のカチオン性脂質

またはその薬学的に許容可能な塩。
３１．式（ＩＩＧ）に従う構造を有する番号付けされた実施形態２７に記載のカチオン性脂質

またはその薬学的に許容可能な塩。
３２．式（ＩＩＨ）に従う構造を有する番号付けされた実施形態２７に記載のカチオン性脂質であって、

式中、ＹおよびＺのうちの一つはＯＨであり、他方は－ＯＣ（Ｏ）Ｒ’であるか、またはＹおよびＺの両方は、それぞれ独立して－ＯＣ（Ｏ）Ｒ’であるカチオン性脂質、またはその薬学的に許容可能な塩。
３３．式（ＩＩＩ）に従う構造を有する番号付けされた実施形態１～２６のいずれか一つに記載のカチオン性脂質：

またはその薬学的に許容可能な塩。
３４．式（ＩＩＩＡ）に従う構造を有する番号付けされた実施形態３３に記載のカチオン性脂質

またはその薬学的に許容可能な塩。
３５．式（ＩＩＩＢ）に従う構造を有する番号付けされた実施形態３３または番号付けされた実施形態３４に記載のカチオン性脂質

またはその薬学的に許容可能な塩。
３６．式（ＩＩＩＣ）に従う構造を有する番号付けされた実施形態３５に記載のカチオン性脂質

またはその薬学的に許容可能な塩。
３７．式（ＩＩＩＤ）に従う構造を有する番号付けされた実施形態３３に記載のカチオン性脂質

またはその薬学的に許容可能な塩。
３８．式（ＩＩＩＥ）、（ＩＩＩＦ）、（ＩＩＩＧ）、（ＩＩＩＨ）、（ＩＩＩＩ）、（ＩＩＩＪ）、または（ＩＩＩＫ）から選択される構造を有する、番号付けされた実施形態３７に記載のカチオン性脂質

またはその薬学的に許容可能な塩。
３９．式（ＩＩＩＬ）に従う構造を有する番号付けされた実施形態３３に記載のカチオン性脂質

またはその薬学的に許容可能な塩。
４０．式（ＩＶ）に従う構造を有する番号付けされた実施形態３３に記載のカチオン性脂質であって、

式中、Ｍは、Ｈ、ＯＨ、ＯＭｅもしくはＭｅから選択されるカチオン性脂質、またはその薬学的に許容可能な塩。
４１．式（ＶＩ）、（ＶＩＩ）、（ＶＩＩＩ）、（ＩＸ）または（Ｘ）に従う構造を有する番号付けされた実施形態３３に記載のカチオン性脂質であって、

式中、ＹおよびＺのうちの一つはＯＨであり、他方は－ＯＣ（Ｏ）Ｒ’であるか、またはＹおよびＺの両方は、それぞれ独立して－ＯＣ（Ｏ）Ｒ’であるカチオン性脂質、またはその薬学的に許容可能な塩。
４２．ＹおよびＺのうちの一つがＯＨであり、他方が－ＯＣ（Ｏ）Ｒ’である、番号付けされた実施形態４１に記載のカチオン性脂質またはその薬学的に許容可能な塩。
４３．ＹがＯＨであり、Ｚが－ＯＣ（Ｏ）Ｒ’である、番号付けされた実施形態４２に記載のカチオン性脂質またはその薬学的に許容可能な塩。
４４．Ｙが－ＯＣ（Ｏ）Ｒ’であり、ＺがＯＨである、番号付けされた実施形態４２に記載のカチオン性脂質またはその薬学的に許容可能な塩。
４５．ＹおよびＺの両方が－ＯＣ（Ｏ）Ｒ’である、番号付けされた実施形態４１に記載のカチオン性脂質またはその薬学的に許容可能な塩。
４６．表１～８に列挙されたものから選択される化合物、またはその薬学的に許容可能な塩。
４７．番号付けされた実施形態１～４６のいずれか一つに記載のカチオン性脂質、一つまたは複数の非カチオン性脂質、一つまたは複数のコレステロール系脂質、および一つまたは複数のＰＥＧ修飾脂質を含む組成物。
４８．組成物が、脂質ナノ粒子、任意選択的にリポソームである、番号付けされた実施形態４７に記載の組成物。
４９．一つまたは複数のカチオン性脂質が、脂質ナノ粒子の約３０モル％～６０モル％を構成する、番号付けされた実施形態４８に記載の組成物。
５０．一つまたは複数の非カチオン性脂質が、脂質ナノ粒子の約１０モル％～５０モル％を構成する、番号付けされた実施形態４８または４９のいずれか一つに記載の組成物。
５１．一つまたは複数のＰＥＧ修飾脂質が、脂質ナノ粒子の約１モル％～１０モル％を構成する、番号付けされた実施形態４８～５０のいずれか一つに記載の組成物。
５２．コレステロール系脂質が、脂質ナノ粒子の約１０モル％～５０モル％を構成する、番号付けされた実施形態４８～５１のいずれか一つに記載の組成物。
５３．脂質ナノ粒子が、核酸、任意選択的にペプチドまたはタンパク質をコードするｍＲＮＡを封入する、番号付けされた実施形態４８～５２のいずれか一つに記載の組成物。
５４．脂質ナノ粒子が、ペプチドまたはタンパク質をコードするｍＲＮＡを封入する、番号付けされた実施形態４８～５２のいずれか一つに記載の組成物。
５５．脂質ナノ粒子が、少なくとも７０％のｍＲＮＡの封入化率を有する、番号付けされた実施形態５４に記載の組成物。
５６．脂質ナノ粒子が、少なくとも７５％のｍＲＮＡの封入化率を有する、番号付けされた実施形態５４に記載の組成物。
５７．脂質ナノ粒子が、少なくとも８０％のｍＲＮＡの封入化率を有する、番号付けされた実施形態５４に記載の組成物。
５８．脂質ナノ粒子が、少なくとも８５％のｍＲＮＡの封入化率を有する、番号付けされた実施形態５４に記載の組成物。
５９．脂質ナノ粒子が、少なくとも９０％のｍＲＮＡの封入化率を有する、番号付けされた実施形態５４に記載の組成物。
６０．脂質ナノ粒子が、少なくとも９５％のｍＲＮＡの封入化率を有する、番号付けされた実施形態５４に記載の組成物。
６１．療法で使用するための番号付けされた実施形態５４～６０のいずれか一つに記載の組成物。
６２．任意選択的に、疾患が、（ａ）タンパク質欠損症、任意選択的に、タンパク質欠損症が、肝臓、肺、脳または筋肉に影響を及ぼす、（ｂ）自己免疫性疾患、（ｃ）感染症、または（ｄ）癌である、ｍＲＮＡによってコードされるペプチドまたはタンパク質による治療または予防に適している疾患を治療または予防する方法で使用するための番号付けされた実施形態５４～６０のいずれか一つに記載の組成物。
６３．組成物が、静脈内、くも膜下腔内、または筋肉内、または肺送達によって、任意選択的に噴霧を介して投与される、番号付けされた実施形態６１または６２による使用のための組成物。
６４．疾患を治療または予防するための方法であって、該方法が、番号付けされた実施形態５４～６０のいずれか一つに記載の組成物を、それを必要とする対象に投与することを含み、疾患が、ｍＲＮＡによってコードされるペプチドまたはタンパク質による治療または予防に適しており、任意選択的に、疾患が、（ａ）タンパク質欠損症、任意選択的に、タンパク質欠損症が、肝臓、肺、脳または筋肉に影響を及ぼす、（ｂ）自己免疫性疾患、（ｃ）感染症、または（ｄ）癌である、方法。
６５．組成物が、静脈内、くも膜下腔内、または筋肉内、または肺送達によって、任意選択的に噴霧を介して投与される、番号付けされた実施形態６４に記載の方法。 Numbered Embodiments A cationic lipid having a structure according to formula (I),

wherein L ₁ is a bond, (C ₁ -C ₆ )alkyl or (C ₂ -C ₆ )alkenyl;
wherein X is O or S;
wherein R ¹ , R ² , R ³ , R ⁴ and R ⁵ are each independently H, OH, optionally substituted (C ₁ -C ₆ )alkyl, optionally substituted (C ₂ - C ₆ )alkenyl, optionally substituted (C ₂ -C ₆ )alkynyl, optionally substituted (C ₁ -C ₆ )alkoxy and —OC(O)R′;
wherein at least one of R ¹ , R ² , R ³ , R ⁴ or R ⁵ is —OC(O)R′;
wherein R' is

wherein ^R6 is

wherein m and p are each independently 0, 1, 2, 3, 4, or 5;
wherein R ⁷ is H, optionally substituted (C ₁ -C ₆ )alkyl, optionally substituted (C ₂ -C ₆ )alkenyl, optionally substituted (C ₂ -C ₆ )alkynyl , optionally substituted (C ₁ -C ₆ )acyl, —(CH ₂ ) _k R ^A or —(CH ₂ ) _k CH(OR ¹¹ )R ^A ;
wherein R ⁸ is H, optionally substituted (C ₁ -C ₆ )alkyl, optionally substituted (C ₂ -C ₆ )alkenyl, optionally substituted (C ₂ -C ₆ )alkynyl , optionally substituted (C ₁ -C ₆ )acyl, —(CH ₂ ) _n R ^B or —(CH ₂ ) _n CH(OR ¹² )R ^B ;
wherein R ⁹ is H, optionally substituted (C ₁ -C ₆ )alkyl, optionally substituted (C ₂ -C ₆ )alkenyl, optionally substituted (C ₂ -C ₆ )alkynyl , optionally substituted (C ₁ -C ₆ )acyl, —(CH ₂ ) _q R ^C or —(CH ₂ ) _q CH(OR ¹³ )R ^C ;
wherein R ¹⁰ is H, optionally substituted (C ₁ -C ₆ )alkyl, optionally substituted (C ₂ -C ₆ )alkenyl, optionally substituted (C ₂ -C ₆ )alkynyl , optionally substituted (C ₁ -C ₆ )acyl, —(CH ₂ ) _r R ^D or —(CH ₂ ) _r CH(OR ¹⁴ )R ^D ;
wherein k, n, q and r are each independently 1, 2, 3, 4 or 5;
or wherein (i) R ⁷ and R ⁸ or (ii) R ⁹ and R ¹⁰ together form an optionally substituted 5- or 6-membered heterocycloalkyl or heteroaryl, heterocycloalkyl or heterocycloalkyl aryl contains 1 to 3 heteroatoms selected from N, O and S;
wherein R ¹¹ , R ¹² , R ¹³ and R ¹⁴ are each independently selected from H, methyl, ethyl or propyl;
wherein R ^A , R ^B , R ^C and R ^D are each independently optionally substituted (C ₆ -C ₂₀ )alkyl, optionally substituted (C ₆ -C ₂₀ )alkenyl, optionally substituted (C ₆ -C ₂₀ )alkynyl, optionally substituted (C ₆ -C ₂₀ )acyl, optionally substituted —OC(O)alkyl, optionally substituted —OC(O)alkenyl, optionally substituted (C ₁ -C ₆ )monoalkylamino, optionally substituted (C ₁ -C ₆ )dialkylamino, optionally substituted (C ₁ -C ₆ )alkoxy, —OH, —NH selected from ₂ ,
wherein at least one of R ⁷ , R ⁸ , R ⁹ , R ¹⁰ each comprises a R ^A , R ^B , R ^C or R ^D moiety and R ^A , R ^B , R ^C or R ^D is Independently optionally substituted (C ₆ -C ₂₀ )alkyl, optionally substituted (C ₆ -C ₂₀ )alkenyl, optionally substituted (C ₆ -C ₂₀ )alkynyl, optionally substituted (C ₆ -C ₂₀ )acyl, optionally substituted —OC(O)(C ₆ -C ₂₀ )alkyl or optionally substituted —OC(O)(C ₆ -C ₂₀ )alkenyl a cationic lipid that
or a pharmaceutically acceptable salt thereof.
2. Any alkyl, alkenyl, alkynyl, acyl, alkoxy, monoalkylamino, dialkylamino, heterocycloalkyl or heteroaryl may be (C ₁ -C ₆ )alkyl, (C ₂ -C ₆ )alkenyl, (C ₂ -C ₆ ) alkynyl, (C ₁ -C ₆ )acyl, (C ₁ -C ₆ )alkoxy, halogen, —COR, —CO ₂ H, —CO ₂ R, —CN, —OH, —OR, —OCOR, — optionally substituted with one or more substituents selected from the group consisting of OCO ₂ R, —NH ₂ , —NHR, —N(R) ₂ , —SR or —SO ₂ R; 2. The _cationic lipid according to numbered embodiment 1, _or a pharmaceutically acceptable acceptable salt.
3.
3. The cationic lipid according to any of numbered embodiments 1-2, or a cationic lipid according to any of numbered embodiments 1-2, wherein i) R ^A and R ^B are the same and/or ii) R ^C and R ^D are the same; acceptable salt.
4.
The cationic lipid according to numbered embodiments 1 or 2, or a pharmaceutically acceptable salt thereof, wherein i) R ^A and R ^B are different and/or ii) R ^C and R ^D are different.
5. The cationic lipid according to any one of numbered embodiments 1-3, or a pharmaceutically acceptable salt thereof, wherein R ^A , R ^B , R ^C and R ^D are the same.
6. The cationic lipid according to any one of numbered embodiments 1-4, or a pharmaceutically acceptable salt thereof, according to any one of numbered embodiments 1-4, wherein one or more of R ^A , R ^B , R ^C and R ^D are different .
7. The cationic lipid according to any one of numbered embodiments 1-6, wherein X is O, or a pharmaceutically acceptable salt thereof.
8. The cationic lipid according to any one of numbered embodiments 1-6, wherein X is S, or a pharmaceutically acceptable salt thereof.
9. Cationic according to any one of numbered embodiments 1-8, wherein only one of R ¹ , R ² , R ³ , R ⁴ and R ⁵ is —OC(O)R′ A lipid or a pharmaceutically acceptable salt thereof.
10. The cationic lipid according to numbered embodiment 9, or a pharmaceutically acceptable salt thereof, according to numbered embodiment 9, wherein none of R ¹ , R ² , R ³ , R ⁴ or R ⁵ is OH.
11. The cationic lipid according to any one of numbered embodiments 1-8, wherein two of R ¹ , R ² , R ³ , R ⁴ and R ⁵ are —OC(O)R′ or a pharmaceutically acceptable salt thereof.
12. The cationic lipid according to numbered embodiment 11, or a pharmaceutically acceptable salt thereof, according to numbered embodiment 11, wherein none of R ¹ , R ² , R ³ , R ⁴ or R ⁵ is OH.
13. The cationic lipid according to any one of numbered embodiments 1-8, wherein three of R ¹ , R ² , R ³ , R ⁴ and R ⁵ are —OC(O)R′ or a pharmaceutically acceptable salt thereof.
14. The cationic lipid according to any one of numbered embodiments 1-13, or a pharmaceutically acceptable salt thereof, wherein R ¹ and/or R ⁵ are —OC(O)R′.
15. The cationic lipid according to any one of numbered embodiments 1-14, or a pharmaceutically acceptable salt thereof, wherein R ² and/or R ⁴ are —OC(O)R′.
16. The cationic lipid according to any one of numbered embodiments 1-15, or a pharmaceutically acceptable salt thereof, wherein R ³ is -OC(O)R'.
17.
i) p, q, and r are the same, or ii) one or more of p, q, and r are different, or iii) q and r are the same and p is different, numbered A cationic lipid according to any one of embodiments 1-16 or a pharmaceutically acceptable salt thereof.
18.
i) k, m, and n are the same, or ii) one or more of k, m and n are different, or iii) k and n are the same and m is different, numbered A cationic lipid according to any one of embodiments 1-17 or a pharmaceutically acceptable salt thereof.
19. The cationic lipid according to any one of numbered embodiments 1-18, wherein m is 1, 2, or 3, or a pharmaceutically acceptable salt thereof.
20. The cationic lipid according to any one of numbered embodiments 1-19, wherein p is 1, 2, or 3, or a pharmaceutically acceptable salt thereof.
21. The cationic lipid according to any one of numbered embodiments 1-20, or a pharmaceutically acceptable salt thereof, wherein R ^' is:

22.
i) k, m and n = 1, or ii) k, m and n = 1 and ^R11 and ^R12 = H, or iii) k and n = 1 and m = 2, or iv) k and n = 1, m = 2, and ^R11 and ^R12 = H, or v) k and n = 1, and m = 3, or vi) k and n = 1, m = 3, and ^R11 and ^R12 = 22. The cationic lipid according to numbered embodiment 21, which is H, or a pharmaceutically acceptable salt thereof.
23. The cationic lipid according to any one of numbered embodiments 1-22, or a pharmaceutically acceptable salt thereof, wherein R ⁶ is:

24.
i) p, q and r=1, or ii) p, q and r=1 and ^R13 and ^R14 are H, or iii) q and r=1 and p=2, or iv) q and r=1, p=2, and ^R13 and ^R14 are H. The cationic lipid according to numbered embodiment 23, or a pharmaceutically acceptable salt thereof.
25. The cationic lipid according to any one of numbered embodiments 1-22, or a pharmaceutically acceptable salt thereof, wherein R ⁶ is selected from the group consisting of:

26. 26. The cationic lipid according to numbered embodiment 25, or a pharmaceutically acceptable salt thereof, wherein ^R6 is:

27. The cationic lipid according to any one of numbered embodiments 1-26 having a structure according to Formula (II):

or a pharmaceutically acceptable salt thereof.
28. The cationic lipid according to numbered embodiment 27 having a structure according to Formula (IIA)

or a pharmaceutically acceptable salt thereof.
29. 29. The cationic lipid according to numbered embodiment 28, having a structure according to one of formulas (IIB), (IIC), (IID), or (IIE).

or a pharmaceutically acceptable salt thereof.
30. The cationic lipid according to numbered embodiment 27 having a structure according to Formula (IIF)

or a pharmaceutically acceptable salt thereof.
31. The cationic lipid according to numbered embodiment 27 having a structure according to Formula (IIG)

or a pharmaceutically acceptable salt thereof.
32. 28. The cationic lipid according to numbered embodiment 27 having a structure according to formula (IIH),

wherein one of Y and Z is OH and the other is -OC(O)R' or both Y and Z are each independently -OC(O)R' A cationic lipid, or a pharmaceutically acceptable salt thereof.
33. The cationic lipid according to any one of numbered embodiments 1-26 having a structure according to Formula (III):

or a pharmaceutically acceptable salt thereof.
34. 34. The cationic lipid according to numbered embodiment 33, having a structure according to Formula (IIIA)

or a pharmaceutically acceptable salt thereof.
35. The cationic lipid according to numbered embodiment 33 or numbered embodiment 34 having a structure according to Formula (IIIB)

or a pharmaceutically acceptable salt thereof.
36. The cationic lipid according to numbered embodiment 35 having a structure according to Formula (IIIC)

or a pharmaceutically acceptable salt thereof.
37. 34. The cationic lipid according to numbered embodiment 33, having a structure according to Formula (IIID)

or a pharmaceutically acceptable salt thereof.
38. 38. The cationic lipid according to numbered embodiment 37, having a structure selected from formulas (IIIE), (IIIF), (IIIG), (IIIH), (III), (IIIJ), or (IIIK)

or a pharmaceutically acceptable salt thereof.
39. 34. The cationic lipid according to numbered embodiment 33, having a structure according to Formula (IIIL)

or a pharmaceutically acceptable salt thereof.
40. 34. The cationic lipid according to numbered embodiment 33 having a structure according to formula (IV),

wherein M is a cationic lipid selected from H, OH, OMe or Me, or a pharmaceutically acceptable salt thereof.
41. 34. The cationic lipid according to numbered embodiment 33, having a structure according to formula (VI), (VII), (VIII), (IX) or (X),

wherein one of Y and Z is OH and the other is -OC(O)R' or both Y and Z are each independently -OC(O)R' A cationic lipid, or a pharmaceutically acceptable salt thereof.
42. 42. The cationic lipid according to numbered embodiment 41, or a pharmaceutically acceptable salt thereof, according to numbered embodiment 41, wherein one of Y and Z is OH and the other is -OC(O)R'.
43. 43. The cationic lipid according to numbered embodiment 42, or a pharmaceutically acceptable salt thereof, wherein Y is OH and Z is -OC(O)R'.
44. 43. The cationic lipid according to numbered embodiment 42, or a pharmaceutically acceptable salt thereof, wherein Y is -OC(O)R' and Z is OH.
45. 42. The cationic lipid according to numbered embodiment 41, or a pharmaceutically acceptable salt thereof, wherein both Y and Z are -OC(O)R'.
46. A compound selected from those listed in Tables 1-8, or a pharmaceutically acceptable salt thereof.
47. a cationic lipid, one or more non-cationic lipids, one or more cholesterol-based lipids, and one or more PEG-modified lipids according to any one of numbered embodiments 1-46 A composition comprising:
48. 48. The composition according to numbered embodiment 47, wherein the composition is a lipid nanoparticle, optionally a liposome.
49. 49. The composition of numbered embodiment 48, wherein the one or more cationic lipids constitute about 30-60 mol % of the lipid nanoparticles.
50. 50. The composition according to any one of numbered embodiments 48 or 49, wherein the one or more non-cationic lipids constitute about 10 mol % to 50 mol % of the lipid nanoparticles.
51. 51. The composition according to any one of numbered embodiments 48-50, wherein the one or more PEG-modified lipids constitute about 1-10 mol% of the lipid nanoparticles.
52. 52. The composition of any one of numbered embodiments 48-51, wherein the cholesterol-based lipid comprises about 10-50 mol% of the lipid nanoparticles.
53. 53. The composition of any one of numbered embodiments 48-52, wherein the lipid nanoparticles encapsulate mRNA encoding nucleic acids, optionally peptides or proteins.
54. 53. The composition of any one of numbered embodiments 48-52, wherein the lipid nanoparticles encapsulate mRNA encoding the peptide or protein.
55. 55. The composition of numbered embodiment 54, wherein the lipid nanoparticles have an encapsulation rate of mRNA of at least 70%.
56. 55. The composition of numbered embodiment 54, wherein the lipid nanoparticles have an encapsulation rate of mRNA of at least 75%.
57. 55. The composition of numbered embodiment 54, wherein the lipid nanoparticles have an encapsulation rate of mRNA of at least 80%.
58. 55. The composition of numbered embodiment 54, wherein the lipid nanoparticles have an encapsulation rate of mRNA of at least 85%.
59. 55. The composition of numbered embodiment 54, wherein the lipid nanoparticles have an encapsulation rate of mRNA of at least 90%.
60. 55. The composition of numbered embodiment 54, wherein the lipid nanoparticles have an encapsulation rate of mRNA of at least 95%.
61. A composition according to any one of numbered embodiments 54-60 for use in therapy.
62. Optionally, the disease is (a) a protein deficiency, optionally the protein deficiency affects the liver, lungs, brain or muscle, (b) an autoimmune disease, (c) an infectious disease or (d) cancer, any one of numbered embodiments 54-60 for use in a method of treating or preventing a disease amenable to treatment or prevention with a peptide or protein encoded by the mRNA. The composition according to 1.
63. A composition for use according to numbered embodiments 61 or 62, wherein the composition is administered by intravenous, intrathecal, or intramuscular, or pulmonary delivery, optionally via nebulization.
64. A method for treating or preventing a disease comprising administering a composition according to any one of numbered embodiments 54-60 to a subject in need thereof. , the disease is suitable for treatment or prevention with a peptide or protein encoded by the mRNA, optionally the disease is (a) a protein deficiency, optionally the protein deficiency is liver, lung, The method is (b) an autoimmune disease, (c) an infectious disease, or (d) cancer that affects the brain or muscles.
65. 65. The method according to numbered embodiment 64, wherein the composition is administered by intravenous, intrathecal, or intramuscular, or pulmonary delivery, optionally via nebulization.

Claims (15)

A cationic lipid having a structure according to formula (I),

wherein L ₁ is a bond, (C ₁ -C ₆ )alkyl or (C ₂ -C ₆ )alkenyl;
wherein X is O or S;
wherein R ¹ , R ² , R ³ , R ⁴ and R ⁵ are each independently H, OH, optionally substituted (C ₁ -C ₆ )alkyl, optionally substituted (C ₂ - C ₆ )alkenyl, optionally substituted (C ₂ -C ₆ )alkynyl, optionally substituted (C ₁ -C ₆ )alkoxy and —OC(O)R′;
wherein at least one of R ¹ , R ² , R ³ , R ⁴ or R ⁵ is —OC(O)R′;
wherein R' is

wherein ^R6 is

wherein m and p are each independently 0, 1, 2, 3, 4, or 5;
wherein R ⁷ is H, optionally substituted (C ₁ -C ₆ )alkyl, optionally substituted (C ₂ -C ₆ )alkenyl, optionally substituted (C ₂ -C ₆ )alkynyl , optionally substituted (C ₁ -C ₆ )acyl, —(CH ₂ ) _k R ^A or —(CH ₂ ) _k CH(OR ¹¹ )R ^A ;
wherein R ⁸ is H, optionally substituted (C ₁ -C ₆ )alkyl, optionally substituted (C ₂ -C ₆ )alkenyl, optionally substituted (C ₂ -C ₆ )alkynyl , optionally substituted (C ₁ -C ₆ )acyl, —(CH ₂ ) _n R ^B or —(CH ₂ ) _n CH(OR ¹² )R ^B ;
wherein R ⁹ is H, optionally substituted (C ₁ -C ₆ )alkyl, optionally substituted (C ₂ -C ₆ )alkenyl, optionally substituted (C ₂ -C ₆ )alkynyl , optionally substituted (C ₁ -C ₆ )acyl, —(CH ₂ ) _q R ^C or —(CH ₂ ) _q CH(OR ¹³ )R ^C ;
wherein R ¹⁰ is H, optionally substituted (C ₁ -C ₆ )alkyl, optionally substituted (C ₂ -C ₆ )alkenyl, optionally substituted (C ₂ -C ₆ )alkynyl , optionally substituted (C ₁ -C ₆ )acyl, —(CH ₂ ) _r R ^D or —(CH ₂ ) _r CH(OR ¹⁴ )R ^D ;
wherein k, n, q and r are each independently 1, 2, 3, 4 or 5;
or wherein (i) R ⁷ and R ⁸ or (ii) R ⁹ and R ¹⁰ together form an optionally substituted 5- or 6-membered heterocycloalkyl or heteroaryl, heterocycloalkyl or heterocycloalkyl aryl contains 1 to 3 heteroatoms selected from N, O and S;
wherein R ¹¹ , R ¹² , R ¹³ and R ¹⁴ are each independently selected from H, methyl, ethyl or propyl;
wherein R ^A , R ^B , R ^C and R ^D are each independently optionally substituted (C ₆ -C ₂₀ )alkyl, optionally substituted (C ₆ -C ₂₀ )alkenyl, optionally substituted (C ₆ -C ₂₀ )alkynyl, optionally substituted (C ₆ -C ₂₀ )acyl, optionally substituted —OC(O)alkyl, optionally substituted —OC(O)alkenyl, optionally substituted (C ₁ -C ₆ )monoalkylamino, optionally substituted (C ₁ -C ₆ )dialkylamino, optionally substituted (C ₁ -C ₆ )alkoxy, —OH, —NH selected from ₂ ,
wherein at least one of R ⁷ , R ⁸ , R ⁹ , R ¹⁰ each comprises a R ^A , R ^B , R ^C or R ^D moiety and R ^A , R ^B , R ^C or R ^D is Independently optionally substituted (C ₆ -C ₂₀ )alkyl, optionally substituted (C ₆ -C ₂₀ )alkenyl, optionally substituted (C ₆ -C ₂₀ )alkynyl, optionally substituted (C ₆ -C ₂₀ )acyl, optionally substituted —OC(O)(C ₆ -C ₂₀ )alkyl or optionally substituted —OC(O)(C ₆ -C ₂₀ )alkenyl a cationic lipid that
or a pharmaceutically acceptable salt thereof. 2. The cationic lipid of claim 1, wherein X is O, or a pharmaceutically acceptable salt thereof. 3. The cationic lipid or pharmaceutically acceptable salt thereof according to claim 1 or claim 2, wherein m is 1, 2 or 3. 4. The cationic lipid or pharmaceutically acceptable salt thereof according to any one of claims 1-3, wherein p is 1, 2 or 3. 5. The cationic lipid according to any one of claims 1 to 4, or a pharmaceutically acceptable salt thereof, wherein R ^' is

vii) k, m and n = 1, or viii) k, m and n = 1, and ^R11 and ^R12 = H, or ix) k and n = 1, and m = 2, or x) k and n = 1, m = 2, and ^R11 and ^R12 = H, or xi) k and n = 1, and m = 3, or xii) k and n = 1, m = 3, and ^R11 and ^R12 = 6. The cationic lipid of claim 5, which is H or a pharmaceutically acceptable salt thereof. R' is

and R ⁷ and R ⁸ are each optionally substituted (C ₁ -C ₆ )alkyl substituted with —CO ₂ R ^aa , wherein R ^aa is C ₁ -C ₅₀ alkyl , preferably R ⁷ and R ⁸ are each (C ₁ -C ₆ )alkyl substituted with —CO ₂ R ^aa , wherein R ^aa is C ₁ -C ₂₀ alkyl, more preferably , R ⁷ and R ⁸ are respectively

The cationic lipid according to any one of claims 1 to 4 or a pharmaceutically acceptable salt thereof, which is 8. The cationic lipid according to any one of claims 1 to 7, or a pharmaceutically acceptable salt thereof, wherein ^R6 is:

A cationic lipid having a structure according to
(a) Formula (II):

or a pharmaceutically acceptable salt thereof,
(b) Formula (IIA):

or a pharmaceutically acceptable salt thereof,
(c) Formula (IIF):

or a pharmaceutically acceptable salt thereof,
(d) Formula (IIG):

or a pharmaceutically acceptable salt thereof,
(e) Formula (IIH):

wherein one of Y and Z is OH and the other is -OC(O)R' or both Y and Z are each independently -OC(O)R' a cationic lipid, or a pharmaceutically acceptable salt thereof;
(f) Formula (III):

or a pharmaceutically acceptable salt thereof,
(g) Formula (IIIA):

or a pharmaceutically acceptable salt thereof,
(h) Formula (IIIB):

or a pharmaceutically acceptable salt thereof,
(i) Formula (IIID):

or a pharmaceutically acceptable salt thereof,
(j) Formula (IIIL):

or a pharmaceutically acceptable salt thereof, or (k) Formula (IV):

A cationic lipid according to any one of claims 1 to 8, wherein M is a cationic lipid selected from H, OH, OMe or Me, or a pharmaceutically acceptable salt thereof. A cationic lipid having a structure according to formula (VI), (VII), (VIII), (IX) or (X),

a cation wherein one of Y and Z is OH and the other is -OC(O)R', or both Y and Z are each independently -OC(O)R' The cationic lipid according to any one of claims 1 to 8, which is a cationic lipid, or a pharmaceutically acceptable salt thereof. A compound selected from those listed in Tables 1-8, or a pharmaceutically acceptable salt thereof. A composition comprising a cationic lipid according to any one of claims 1 to 11, one or more non-cationic lipids, one or more cholesterol-based lipids, and one or more PEG-modified lipids A composition, optionally wherein the composition is a lipid nanoparticle. 13. The composition of claim 12, wherein said composition is a lipid nanoparticle, said lipid nanoparticle encapsulating mRNA encoding a nucleic acid, optionally a peptide or protein. 14. The composition of claim 13, wherein the lipid nanoparticles have an encapsulation rate of mRNA of at least 70%. A composition according to any one of claims 12-14 for use in therapy.

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