Nanomaterials containing ionizable lipids

Abstract

The present disclosure describes compositions, preparations, nanoparticles (such as lipid nanoparticles), and / or nanomaterials, and methods of using the same.

Description

[Technical field]

[0001] CROSS-REFERENCE TO RELATED APPLICATIONS This application claims the benefit of U.S. Provisional Application No. 63 / 139,732, filed January 20, 2021, the entire contents of which are incorporated herein by reference. [Background technology]

[0002] In chemistry, biology, and medicine, drug delivery systems have been problematic and have been hampered by a lack of understanding of how the molecular properties of the systems control delivery to tissues and confer drug efficacy. Summary of the Invention

[0003] The present invention recognizes the need for compositions, preparations, nanoparticles, and / or nanomaterials, and methods of using the same. In particular, the present disclosure recognizes that the structural features of compositions, preparations, nanoparticles, and / or nanomaterials affect functional responses in vivo, in vitro, and ex vivo (e.g., through an effect on one or more characteristics, such as, for example, biocompatibility, degradation, manufacturability, stability, tropism, etc.).

[0004] The present disclosure provides certain insight that, in some embodiments, compositions, preparations, nanoparticles, and / or nanomaterials may benefit from the inclusion of one or more lipids having a reduced number of stereocenters compared to certain alternative lipid compounds (e.g., comparable lipid compounds having a core portion that includes one or more stereocenters). In some embodiments, one or more lipids are provided and / or utilized that have a core portion that is substantially free of or completely devoid of stereocenters.

[0005] In particular, the present disclosure recognizes that advantages provided by reducing stereochemistry can include easier synthesis, higher purity, and / or consistency of lipid compound preparations and / or compositions, preparations, nanoparticles, and / or nanomaterials comprising them. For example, in some embodiments, benefits derive from improved homogeneity of the compounds prepared and / or utilized, and / or omission of one or more stereoisomer separation steps.

[0006] Alternatively or additionally, in some embodiments, the advantage may include higher homogeneity of biological and / or functional characteristics.For example, the present disclosure provides insight that biological (e.g., in cells, tissues, or organisms) mechanisms that interact with one or more lipids and / or compositions, preparations, nanoparticles, and / or nanomaterials that contain them may interact differently and / or have a preference for one or more stereoisomers compared to other stereoisomers.According to the present disclosure, therefore, preparations with higher stereoisomeric homogeneity may improve comparability, reliability, and / or efficacy.

[0007] Furthermore, the present disclosure describes, inter alia, that the selection and combination of one or more components described herein affects the functional activity of lipid nanoparticles. In some embodiments, for example, functional activity may refer to desired tropism, stabilization, and / or drug delivery effectiveness. In some embodiments, inter alia, the present disclosure describes that the ratio of one of more components affects one or more functional activities of the compositions, preparations, nanoparticles, and / or nanomaterials described herein.

[0008] Additionally, the present disclosure recognizes that the chemical structure of the lipid confers improved characteristics (e.g., efficacy, stability, biocompatibility, etc.) compared to conventional lipid structures known in the art. For example, the present disclosure provides compounds of formula I: [ka] or a pharma- ceutically acceptable salt thereof, wherein: Each L 1 and L 1’ is independently -C(O)- or -C(O)O-; Each L 2 and L 2’ are independently a covalent bond, an optionally substituted divalent saturated or unsaturated linear or branched C 1 -C 12 a hydrocarbon chain, or [ka] and Each Cy A is independently an optionally substituted ring selected from phenylene, and a 3- to 7-membered saturated or partially unsaturated carbocyclene; each m is independently 0, 1, or 2; Each L 3 and L 3’ are independently a covalent bond, —C(O)O—, —OC(O)—, —O—, or —OC(O)O—; Each R 1 and R 1’ is independently an optionally substituted group, in which 1 to 3 methylene units are optionally and independently replaced by -O- or -NR-, and 1 -C 20 Hydrocarbon chains; 3- to 12-membered saturated or partially unsaturated carbocyclic rings; 1-adamantyl; 2-adamantyl; sterolyl; phenyl; and [ka] is an optionally substituted group selected from Each L 4 independently, divalent saturated or unsaturated linear or branched C 1 -C 6 is a hydrocarbon chain, Each A 1 and A 2is independently an optionally substituted C 1 -C 20 Aliphatic or -L 5 -R 5 or Or A 1 and A 2 may be taken together with their intervening atoms to form an optionally substituted ring, [ka] During the ceremony, x is selected from 1 or 2; # is L 4 represents the attachment point to Each L 5 independently, divalent saturated or unsaturated linear or branched C 1 -C 20 a hydrocarbon chain in which one to three methylene units are optionally and independently replaced by -O- or -NR-; Each R 5 is independently an optionally substituted group selected from a 5- to 10-membered aryl ring and a 3- to 8-membered carbocyclic ring; X 1 is a covalent bond, -O-, or -NR-; X 2 is a covalent bond or an optionally substituted divalent saturated or unsaturated linear or branched C 1 -C 12 a hydrocarbon chain in which one to three methylene units are optionally and independently replaced by -O- or -NR-; X 3 is hydrogen or -Cy B and Cy B is an optionally substituted ring selected from 3- to 7-membered saturated or partially unsaturated carbocyclyl; phenyl; 3- to 7-membered saturated or partially unsaturated heterocyclyl having 1-3 heteroatoms independently selected from nitrogen, oxygen, and sulfur; and 5- to 6-membered heteroaryl having 1-3 heteroatoms independently selected from nitrogen, oxygen, and sulfur; Each R is independently hydrogen or optionally substituted C 1 -C 6 is an aliphatic group, However, X 3 is hydrogen, R 1 or R 1’ At least one of the [ka] or a pharma- ceutically acceptable salt thereof.

[0009] In particular, the present disclosure recognizes that lipid nanoparticle (LNP) compositions comprising one or more ionizable lipids may exhibit unexpected properties. For example, the present disclosure provides LNP compositions and / or preparations comprising one or more of the disclosed ionizable lipids that impart unexpected tropism.

[0010] In some embodiments, the compositions, preparations, nanoparticles, and / or nanomaterials provided are for use in methods of treating a disease or disorder, delivering, producing a polypeptide, or slowing / halting the progression of a disease or disorder.

[0011] In some embodiments, the compositions, preparations, nanoparticles, and / or nanomaterials provided are for use in a method of manufacturing.

[0012] In some embodiments, the compositions, preparations, nanoparticles, and / or nanomaterials provided are for use in a characterization method.

[0013] Elements (eg, methods) of embodiments incorporating one aspect of the invention may be applied to embodiments incorporating other aspects of the invention, and vice versa. [Brief description of the drawings]

[0014] [Figure 1] 1 shows an exemplary mRNA screening system for LNP preparations according to an embodiment of the present disclosure.

[0015] [Diagram 2] 1 shows an exemplary siRNA screening system for LNP preparations according to an embodiment of the present disclosure.

[0016] [Diagram 3] 1 shows a bar graph depicting the mean diameter (nm) of exemplary LNP preparations (Exemplary Lipid 1, Exemplary Lipid 2, Exemplary Lipid 3, Exemplary Lipid 4, Exemplary Lipid 5).

[0017] [Figure 4] 1 shows a bar graph depicting the average polydispersity (%) of exemplary LNP preparations (Exemplary Lipid 1, Exemplary Lipid 2, Exemplary Lipid 3, Exemplary Lipid 4, Exemplary Lipid 5).

[0018] [Diagram 5] 1 shows a bar graph illustrating potent delivery of exemplary LNP preparations (Exemplary Lipid 1, Exemplary Lipid 2, Exemplary Lipid 3, Exemplary Lipid 4, Exemplary Lipid 5) to the liver.

[0019] [Figure 6] 1 shows a bar graph illustrating potent delivery of exemplary LNP preparations (Exemplary Lipid 1, Exemplary Lipid 2, Exemplary Lipid 3, Exemplary Lipid 4, Exemplary Lipid 5) to the spleen.

[0020] [Figure 7] 1 shows a bar graph illustrating the potent delivery of exemplary LNP preparations (Exemplary Lipid 1, Exemplary Lipid 2, Exemplary Lipid 3, Exemplary Lipid 4, Exemplary Lipid 5) to bone marrow.

[0021] [Figure 8] 1 shows a bar graph illustrating potent delivery of exemplary LNP preparations (Exemplary Lipid 1, Exemplary Lipid 2, Exemplary Lipid 3, Exemplary Lipid 4, Exemplary Lipid 5) to the kidney.

[0022] [Figure 9]1 shows a bar graph illustrating the efficacy of an exemplary LNP preparation (exemplary lipid 6) for delivering hEPO mRNA in a non-human primate (NHP) model. After administration of vehicle (saline control) and exemplary lipid 6, the amount of hEPO (ng / mL) in plasma was measured over various time points (0, 4, 6, 24, 48, 72 hours).

[0023] [Figure 10] 1 shows a bar graph illustrating the tolerability of an exemplary LNP preparation (Exemplary Lipid 6). The amount of IL-6 (pg / mL) in plasma was measured over various time points (2, 4, 6 hours) after administration of vehicle (saline control) and exemplary lipid 6.

[0024] [Figure 11] 1 shows a bar graph illustrating potent delivery of an exemplary LNP preparation (Exemplary Lipid 6) to the liver.

[0025] [Figure 12] 1 shows a bar graph showing base editing in liver cells following delivery of an exemplary LNP preparation (exemplary lipid 6) to the liver.

[0026] definition Administration: As used herein, the term "administration" typically refers to the administration of a composition to a subject or system. Those skilled in the art will recognize the various routes that may be utilized for administration to a subject, e.g., a human, under appropriate circumstances. For example, in some embodiments, administration may be ocular, oral, parenteral, topical, and the like. In some particular embodiments, administration may be bronchial (e.g., by bronchial instillation), buccal, dermal (e.g., may include one or more of topical, such as dermal, intradermal, interdermal, transdermal, etc.), enteral, intra-arterial, ventricular, intra-cisternal, intramanguaneal, intradermal, intragastric, intramedullary, intramuscular, intranasal, intraperitoneal, intrathecal, intravenous, intraventricular, within a particular organ (e.g., intrahepatic), mucosal, nasal, oral, rectal, subcutaneous, sublingual, topical, tracheal (e.g., by intratracheal instillation), vaginal, vitreous, by spray, and the like. In some embodiments, administration involves administration that is intermittent (e.g., multiple doses spaced apart over time) and / or periodic (e.g., individual doses spaced apart by the same period of time). In some embodiments, administration may involve continuous administration (e.g., perfusion) for at least a selected period of time. In some embodiments, pharmaceutical compositions comprising lipid nanoparticles may be formulated for administration via parenteral (intramuscular, intraperitoneal, intravenous (IV), or subcutaneous injection), transdermal (either passively or using iontophoresis or electroporation), or transmucosal (nasal, vaginal, rectal, or sublingual) routes of administration, or by using bioerodible inserts, and may be formulated in dosage forms appropriate for each route of administration.

[0027] Aliphatic: The term "aliphatic" or "aliphatic group," as used herein, refers to a straight-chain (i.e., unbranched) or branched, substituted or unsubstituted hydrocarbon chain that is fully saturated or contains one or more units of unsaturation, or a monocyclic or bicyclic hydrocarbon that is fully saturated or contains one or more units of unsaturation, but is not aromatic (also referred to herein as "carbocycle," "carbocyclic," "alicyclic," or "cycloalkyl") and has one point of attachment to the remainder of the molecule. Unless otherwise specified, an aliphatic group contains 1-6 aliphatic carbon atoms. In some embodiments, an aliphatic group contains 1-5 carbon atoms. In some embodiments, an aliphatic group contains 1-4 carbon atoms. In some embodiments, an aliphatic group contains 1-3 carbon atoms, and in some embodiments, an aliphatic group contains 1-2 carbon atoms. In some embodiments, "carbocyclic" (or "alicyclic" or "carbocycle" or "cycloalkyl") refers to a monocyclic C ring that is fully saturated or contains one or more units of unsaturation, but is not aromatic, and has a single point of attachment to the remainder of the molecule. 3 -C 8 Refers to a hydrocarbon. Suitable aliphatic groups include, but are not limited to, linear or branched, substituted or unsubstituted alkyl, alkenyl, alkynyl groups, as well as hybrids thereof, such as (cycloalkyl)alkyl, (cycloalkenyl)alkyl, or (cycloalkyl)alkenyl.

[0028] Alkenyl: The term “alkenyl” used alone or as part of a larger moiety refers to an alkenyl group having at least one double bond and (unless otherwise specified) 2 to 20, 2 to 18, 2 to 16, 2 to 14, 2 to 12, 2 to 10, 2 to 8, 2 to 6, 2 to 4, or 2 to 3 carbon atoms (e.g., C 2-20 , C 2-18 , C 2-16 , C 2-14 , C 2-12 , C 2-10 , C 2-8 , C 2-6 , C 2-4 , or C 2-3) . Exemplary alkenyl groups include ethenyl, propenyl, butenyl, pentenyl, hexenyl, and heptenyl.

[0029] Alkenylene: The term "alkenylene" refers to a divalent alkenyl group. A substituted alkenylene chain is a polymethylene group containing at least one double bond in which one or more hydrogen atoms are replaced with a substituent. Suitable substituents include those described below for substituted aliphatic groups.

[0030] Alkyl: As used herein, the term "alkyl" is given its ordinary meaning in the art and can include saturated aliphatic groups, including straight chain alkyl groups, branched chain alkyl groups, cycloalkyl (alicyclic) groups, alkyl substituted cycloalkyl groups, and cycloalkyl substituted alkyl groups. In some embodiments, an alkyl has 1-100 carbon atoms. In certain embodiments, a straight chain or branched chain alkyl has about 1-20 carbon atoms in its backbone (e.g., C for straight chain). 1 -C 20 , C for branched chains 2 -C 20 ), or about 1-10 carbon atoms. In some embodiments, cycloalkyl rings have from about 3-10 carbon atoms in their ring structure when such rings are monocyclic or bicyclic, or alternatively about 5, 6 or 7 carbons in the ring structure. In some embodiments, an alkyl group can be a lower alkyl group, where the lower alkyl group has from 1-4 carbon atoms (e.g., C for a straight chain lower alkyl). 1 -C 4 ).

[0031] Alkylenyl: The term "alkylenyl" refers to a divalent alkyl group that is linear (i.e., unbranched) or branched, substituted or unsubstituted. "Alkylenyl" refers to a polymethylene group, i.e., -(CH 2 ) n- and n is a positive integer, preferably 1 to 10, 1 to 9, 1 to 8, 1 to 7, 1 to 6, 1 to 5, 1 to 4, 1 to 3, 1 to 2, 2 to 5, or 4 to 8. Substituted alkylenyl is a polymethylene group in which one or more methylene hydrogen atoms have been replaced with a substituent. Suitable substituents include those described below for substituted aliphatic groups.

[0032] Alkynyl: The term “alkynyl” used alone or as part of a larger moiety refers to an alkynyl group having at least one triple bond and (unless otherwise specified) 2 to 20, 2 to 18, 2 to 16, 2 to 14, 2 to 12, 2 to 10, 2 to 8, 2 to 6, 2 to 4, or 2 to 3 carbon atoms (e.g., C 2-20 , C 2-18 , C 2-16 , C 2-14 , C 2-12 , C 2-10 , C 2-8 , C 2-6 , C 2-4 , or C 2-3 ) refers to an optionally substituted straight or branched chain hydrocarbon group having an alkynyl group. Exemplary alkynyl groups include ethynyl, propynyl, butynyl, pentynyl, hexynyl, and heptynyl.

[0033] Amino Acid: In its broadest sense, as used herein, refers to any compound and / or substance that can be incorporated into a polypeptide chain, for example, through the formation of one or more peptide bonds. In some embodiments, an amino acid has the general structure H 2The amino acid has a NC(H)(R)-COOH. In some embodiments, the amino acid is a naturally occurring amino acid. In some embodiments, the amino acid is a non-natural amino acid, in some embodiments, the amino acid is a D-amino acid, and in some embodiments, the amino acid is an L-amino acid. A "standard amino acid" refers to any of the 20 standard L-amino acids that are typically found in naturally occurring peptides. A "non-standard amino acid" refers to any amino acid other than the standard amino acids, whether it is synthetically prepared or obtained from a natural source. In some embodiments, amino acids, including carboxy- and / or amino-terminal amino acids, in a polypeptide may contain structural modifications compared to the general structure above. For example, in some embodiments, an amino acid may be modified by methylation, amidation, acetylation, pegylation, glycosylation, phosphorylation, and / or substitution (e.g., an amino group, a carboxylic acid group, one or more protons, and / or a hydroxyl group) compared to the general structure. In some embodiments, such modifications may, for example, alter the circulating half-life of a polypeptide containing a modified amino acid compared to a polypeptide containing the same unmodified amino acid. In some embodiments, such modifications do not significantly alter the relevant activity of a polypeptide containing the modified amino acid compared to a polypeptide containing the same unmodified amino acid. As will be clear from the context, in some embodiments, the term "amino acid" may be used to refer to a free amino acid, and in some embodiments, to an amino acid residue of a polypeptide.

[0034] Animal: as used herein refers to any member of the animal kingdom. In some embodiments, "animal" refers to humans of either sex and at any stage of development. In some embodiments, "animal" refers to non-human animals at any stage of development. In certain embodiments, the non-human animals are mammals (e.g., 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 may be transgenic animals, genetically engineered animals, and / or clones.

[0035] Aryl: The term "aryl" refers to an alkyl group having a total of 6 to 14 ring members, e.g., C 6-14 ), where at least one ring in the system is aromatic and each ring in the system contains 3 to 7 ring members. The term "aryl" may be used interchangeably with the term "aryl ring." In some embodiments, "aryl" refers to an aromatic ring system, including but not limited to phenyl, naphthyl, anthracyl, and the like, which may bear one or more substituents. Unless otherwise specified, "aryl" groups are hydrocarbons.

[0036] Approximately: As used herein, the term "approximately" or "about" when applied to one or more values ​​of interest refers to a value similar to a stated reference value. In general, a person skilled in the art familiar with the context will understand the relevant degree of difference encompassed by "about" or "approximately" in that context. In certain embodiments, the term "approximately" or "about" refers to a range of values ​​that falls within 25%, 20%, 19%, 18%, 17%, 16%, 15%, 14%, 13%, 12%, 11%, 10%, 9%, 8%, 7%, 6%, 5%, 4%, 3%, 2%, 1% or less in either direction (greater or less) of the stated reference value (except where such numerical value exceeds 100% of possible values), unless otherwise stated or clear from the context.

[0037] Aptamer: As used herein, the term "aptamer" refers to a macromolecule composed of nucleic acids (e.g., RNA, DNA) that binds tightly to a specific molecular target (e.g., umbrella topology glycans). A particular aptamer can be described by a linear nucleotide sequence and is typically about 15-60 nucleotides in length. Without wishing to be bound by any theory, it is contemplated that the nucleotide chains in an aptamer form intramolecular interactions that fold the molecule into a complex three-dimensional shape that allows the aptamer to bind tightly to the surface of its target molecule. Given the extraordinary diversity of molecular shapes that exist within the field of all possible nucleotide sequences, aptamers can be obtained for a wide range of molecular targets, including proteins and small molecules. In addition to high specificity, a typical aptamer has very high affinity for its target (e.g., picomolar to low nanomolar range affinity for proteins). In many embodiments, aptamers are chemically stable and can be boiled or frozen without loss of activity. Aptamers are synthetic molecules, and therefore amenable to various modifications that can optimize their function for specific applications. For example, aptamers can be modified to dramatically reduce their susceptibility to enzyme degradation in blood for use in in vivo applications. In addition, aptamers can be modified to change their biodistribution or plasma residence time.

[0038] Associated: As used herein, two events or entities are "associated" with each other when the presence, level, degree, type, and / or form of one correlates with that of the other. For example, a particular entity (e.g., a polypeptide, gene signature, metabolite, microorganism) is considered to be associated with a particular disease, disorder, or condition when its presence, level, and / or form correlates with the incidence and / or susceptibility of the disease, disorder, or condition (e.g., across a relevant population). In some embodiments, two or more entities are physically "associated" with each other when they directly or indirectly interact with each other such that they are in and / or maintain physical proximity to each other. In some embodiments, two or more entities that are physically associated with each other are covalently linked to each other, and in some embodiments, two or more entities that are physically associated with each other are not covalently linked to each other but are non-covalently associated, for example, by hydrogen bonds, van der Waals interactions, hydrophobic interactions, magnetism, and combinations thereof.

[0039] Biocompatible: The term "biocompatible," as used herein, refers to a material that, when placed in contact with living tissue, e.g., in vivo, does not cause significant harm to such tissue. In certain embodiments, a material is "biocompatible" if it is not toxic to cells. In certain embodiments, materials are "biocompatible" if their addition to cells in vitro results in 20% or less cell death and / or if their administration in vivo does not induce significant inflammation or other such adverse effects.

[0040] Biodegradable: As used herein, the term "biodegradable" refers to a material that, when introduced into a cell, is broken down (e.g., by cellular mechanisms, such as by enzymatic degradation, hydrolysis, and / or a combination thereof) into components that the cell can reuse or dispose of without significant toxic effects on the cell. In certain embodiments, the components produced by the degradation of a biodegradable material are biocompatible and therefore do not induce significant inflammation and / or other adverse effects in vivo. In some embodiments, biodegradable polymeric materials are broken down into their component monomers. In some embodiments, the degradation of a biodegradable material (including, for example, a biodegradable polymeric material) involves hydrolysis of ester bonds. Alternatively or additionally, in some embodiments, the degradation of a biodegradable material (including, for example, a biodegradable polymeric material) involves cleavage of urethane bonds. Exemplary biodegradable polymers include, for example, polymers of hydroxy acids such as lactic acid and glycolic acid, including, but not limited to, poly(hydroxyl acid), poly(lactic acid) (PLA), poly(glycolic acid) (PGA), poly(lactic-co-glycolic acid) (PLGA), and copolymers with PEG, polyanhydrides, poly(ortho)esters, polyesters, polyurethanes, poly(butyric acid), poly(valeric acid), poly(caprolactone), poly(hydroxyalkanoates, poly(lactide-co-caprolactone), blends and copolymers thereof. Also included are, for example, albumin, Many natural polymers are biodegradable, including proteins such as collagen, gelatin, and prolamines, e.g., zein, and polysaccharides such as alginates, cellulose derivatives, and polyhydroxyalkanoates, e.g., polyhydroxybutyrate blends, and copolymers thereof. Those skilled in the art will know or be able to determine when such polymers are their biocompatible and / or biodegradable derivatives (e.g., when related to the parent polymer by substantially identical structure differing only in the substitution or addition of certain chemical groups known in the art).

[0041] Biologically active: as used herein refers to an observable biological effect or result achieved by an agent or entity of interest. For example, in some embodiments, a specific binding interaction is a biological activity. In some embodiments, modulation (e.g., induction, enhancement, or inhibition) of a biological pathway or event is a biological activity. In some embodiments, the presence or degree of biological activity is assessed through detection of a direct or indirect product produced by the biological pathway or event of interest.

[0042] Biological sample: As used herein, the term "biological sample" typically refers to a sample obtained or derived from a biological source of interest (e.g., tissue, or organism, or cell culture) as described herein. In some embodiments, the source of interest includes an organism, such as an animal or human. In some embodiments, the biological sample is or includes a biological tissue or biological fluid. In some embodiments, the biological sample can be or includes bone marrow, blood; blood cells; ascites; tissue or fine needle biopsy sample; cell-containing body fluid; free floating nucleic acid; sputum; saliva; urine; cerebrospinal fluid, peritoneal fluid; pleural fluid; feces; lymph; gynecological fluid; skin swab; vaginal swab; oral swab; nasal swab; washing or lavage, such as ductal lavage or bronchoalveolar lavage; aspirate; scraping; bone marrow specimen; tissue biopsy specimen; surgical specimen; feces, other body fluids, secretions, and / or excretions; and / or cells therefrom. In some embodiments, the biological sample is or comprises cells obtained from an individual. In some embodiments, the cells obtained are or comprise cells from the individual from whom the sample is obtained. In some embodiments, the sample is a "primary sample" obtained directly from the source of interest by any suitable means. For example, in some embodiments, the primary biological sample is obtained by a method selected from the group consisting of biopsy (e.g., fine needle aspiration or tissue biopsy), surgery, collection of bodily fluids (e.g., blood, lymph, feces, etc.), and the like. In some embodiments, as will be clear from the context, the term "sample" refers to a preparation obtained by processing (e.g., by removing one or more components and / or adding one or more agents) of the primary sample. For example, filtering using a semi-permeable membrane. Such a "processed sample" may include, for example, nucleic acids or proteins extracted from the sample or obtained by subjecting the primary sample to techniques such as amplification or reverse transcription of mRNA, isolation and / or purification of certain components.

[0043] Divalent: As used herein, the term "divalent" refers to a chemical moiety that has two points of attachment. For example, a "divalent C 1-8 (or C 1-6) Saturated or unsaturated straight or branched hydrocarbon chain" refers to straight or branched divalent alkylene, alkenylene, and alkynylene chains as defined herein.

[0044] Cancer: The terms "cancer," "malignancy," "neoplasm," "tumor," and "carcinoma" are used herein to refer to cells that exhibit relatively abnormal, uncontrolled, and / or autonomous proliferation, such that they exhibit an abnormal growth phenotype characterized by a marked loss of control of cell proliferation. In some embodiments, a tumor may be or include cells that are precancerous (e.g., benign), malignant, premetastatic, metastatic, and / or nonmetastatic. The present disclosure specifically identifies certain cancers to which its teachings may be particularly relevant. In some embodiments, the relevant cancer may be characterized as a solid tumor. In some embodiments, the relevant cancer may be characterized as a hematological tumor. In general, examples of different types of cancers known in the art include, for example, hematopoietic cancers including leukemia, lymphoma (Hodgkin's and non-Hodgkin's), myeloma, and myeloproliferative disorders; sarcoma, melanoma, adenoma, cancer of solid tissues, squamous cell carcinoma of the mouth, throat, larynx, and lung, liver cancer, reproductive tract cancers such as prostate, cervical, bladder, uterine, and endometrial cancer, as well as benign lesions such as renal cell carcinoma, bone cancer, pancreatic cancer, skin cancer, cutaneous or intraocular melanoma, endocrine system cancer, thyroid cancer, parathyroid cancer, head and neck cancer, breast cancer, gastrointestinal cancer, and nervous system cancers, papilloma, and the like.

[0045] Carbocyclyl: As used herein, the terms "carbocyclyl", "carbocycle", and "carbocyclic ring" refer to a saturated or partially unsaturated cycloaliphatic monocyclic, bicyclic, or polycyclic ring system as described herein having 3 to 14 members, where the aliphatic ring system is optionally substituted as described herein. Carbocyclic groups include, but are not limited to, cyclopropyl, cyclobutyl, cyclopentyl, cyclopentenyl, cyclohexyl, cyclohexenyl, cycloheptyl, cycloheptenyl, cyclooctyl, cyclooctenyl, norbornyl, adamantyl, and cyclooctadienyl. In some embodiments, "carbocyclyl" (or "alicyclic") refers to an optionally substituted monocyclic C 3 -C 8 A hydrocarbon or optionally substituted C group that is fully saturated or contains one or more units of unsaturation, but is not aromatic, and has a single point of attachment to the remainder of the molecule. 7 -C 10 It refers to a bicyclic hydrocarbon. The term "cycloalkyl" refers to an optionally substituted saturated ring system of about 3 to about 10 ring carbon atoms. In some embodiments, the cycloalkyl group has 3 to 6 carbons. Exemplary monocyclic cycloalkyl rings include cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, and cycloheptyl. The term "cycloalkenyl" refers to an optionally substituted non-aromatic monocyclic or polycyclic ring system containing at least one carbon-carbon double bond and having about 3 to about 10 carbon atoms. Exemplary monocyclic cycloalkenyl rings include cyclopentenyl, cyclohexenyl, and cycloheptenyl.

[0046] Carrier: as used herein, refers to a diluent, adjuvant, excipient, or vehicle that is administered with the composition. In some exemplary embodiments, the carrier can include sterile liquids such as, for example, water and oils, including those of petroleum, animal, vegetable, or synthetic origin, such as, for example, peanut oil, soybean oil, mineral oil, sesame oil, etc. In some embodiments, the carrier is or includes one or more solid components.

[0047] Comparable: As used herein, the term "comparable" refers to two or more agents, entities, circumstances, sets of conditions, etc. that may not be identical to each other, but are sufficiently similar to permit a comparison between them, such that a person skilled in the art would understand that a conclusion can be reasonably drawn based on the observed differences or similarities. In some embodiments, a comparable set of conditions, circumstances, individuals, or populations is characterized by a number of substantially identical characteristics and one or a few characteristics that vary. A person skilled in the art will understand what degree of identity is necessary in any given situation for two or more such agents, entities, circumstances, sets of conditions, etc. to be considered comparable, depending on the context. For example, a person skilled in the art will understand that sets of circumstances, individuals, or populations are comparable to each other if the differences in the results obtained or the phenomena observed under or with the different sets of circumstances, individuals, or populations are caused by the variation of those characteristics that vary, or are characterized by a sufficient number and type of substantially identical characteristics to warrant a reasonable conclusion indicating the variation of the characteristics.

[0048] Component: Those skilled in the art will understand that the term "composition" may be used to refer to a separate physical entity that includes one or more defined components. In general, unless otherwise specified, a composition may be in any form, e.g., gas, gel, liquid, solid, etc.

[0049] Comprising: Compositions or methods described herein as "comprising" one or more recited elements or steps are open-ended, meaning that the recited elements or steps are essential, but other elements or steps may be added within the scope of the composition or method. It is also understood that, to avoid redundancy, any composition or method described as "comprising" (or "comprises") one or more recited elements or steps also describes a corresponding, more limited composition or method that "consisting essentially of" (or "consists essentially of") the same recited elements or steps, meaning that the composition or method includes the recited essential elements or steps, and may include additional elements or steps that do not materially affect the basic and novel characteristics of the composition or method. It is also understood that any composition or method described herein as "comprising" or "consisting essentially of" one or more recited elements or steps also describes a corresponding, more limited, close-ended composition or method that "consists of" (or "consists of") the recited elements or steps, to the exclusion of any other unrecited elements or steps. In any composition or method disclosed herein, known or disclosed equivalents of any recited essential elements or steps may be substituted for that element or step.

[0050] "Improve", "Increase", "Inhibit", or "Reduce": As used herein, the terms "improve", "increase", "inhibit", "reduce", or their grammatical equivalents refer to a value compared to a baseline or other reference measurement. In some embodiments, a suitable reference measurement may be or include a measurement in a particular system (e.g., a single individual) under comparable conditions in the absence of a particular agent or treatment (before and / or after), or in the presence of a suitable comparable reference agent. In some embodiments, a suitable reference measurement may be or include a measurement in a comparable system known or expected to respond in a particular manner in the presence of the relevant agent or treatment.

[0051] Determining: Many methodologies described herein include a "determining" step. Upon reading this specification, one of skill in the art will understand that such "determining" may be utilized or accomplished through the use of any of a variety of techniques available to one of skill in the art, including, for example, certain techniques explicitly mentioned herein. In some embodiments, determining involves the manipulation of the physical sample. In some embodiments, determining involves the review and / or manipulation of data or information, for example, using a computer or other processing unit adapted to perform the relevant analysis. In some embodiments, determining involves receiving relevant information and / or materials from a source. In some embodiments, determining involves comparing one or more characteristics of the sample or entity to a comparable reference.

[0052] Encapsulated: The term "encapsulated" is used herein to refer to a substance that is completely surrounded by another material.

[0053] Excipient: as used herein refers to a non-therapeutic agent that may be included in a pharmaceutical composition, for example, to provide or contribute to a desired consistency or stabilizing effect. Suitable pharmaceutical excipients include, for example, starch, glucose, lactose, sucrose, gelatin, malt, rice, flour, chalk, silica gel, sodium stearate, glycerol monostearate, talc, sodium chloride, dried skim milk, glycerol, propylene, glycol, water, ethanol, and the like.

[0054] Expression: As used herein, the term "expression" of a nucleic acid sequence refers to the generation of any gene product from the nucleic acid sequence. In some embodiments, the gene product can be a transcription product. In some embodiments, the gene product can be a polypeptide. In some embodiments, expression of a nucleic acid sequence involves one or more of the following: (1) production of an RNA template from a DNA sequence (e.g., by transcription), (2) processing of the RNA transcript (e.g., by splicing, editing, 5' capping, and / or 3' end formation), (3) translation of the RNA into a polypeptide or protein, and / or (4) post-translational modification of the polypeptide or protein.

[0055] Heteroaryl: The terms "heteroaryl" and "heteroar-" used alone or as part of a larger moiety, such as "heteroaralkyl" or "heteroaralkoxy", refer to monocyclic or bicyclic ring groups having 5 to 10 ring atoms (e.g., 5-6 membered monocyclic heteroaryl or 9-10 membered bicyclic heteroaryl), sharing 6, 10, or 14 pi electrons in the cyclic array, and having 1 to 5 heteroatoms in addition to the carbon atoms. Exemplary heteroaryl groups include, but are not limited to, thienyl, furanyl, pyrrolyl, imidazolyl, pyrazolyl, triazolyl, tetrazolyl, oxazolyl, isoxazolyl, oxadiazolyl, thiazolyl, isothiazolyl, thiadiazolyl, pyridyl, pyridonyl, pyridazinyl, pyrimidinyl, pyrazinyl, indolizinyl, purinyl, naphthyridinyl, pteridinyl, imidazo[1,2-a]pyrimidinyl, imidazo[1,2-a]pyridinyl, thienopyrimidinyl, triazolopyridinyl, and benzisoxazolyl. The terms "heteroaryl" and "heteroar-", as used herein, also include groups in which a heteroaromatic ring is fused to one or more aryl, alicyclic, or heterocyclyl rings, and the radical or point of attachment is on the heteroaromatic ring (i.e., a bicyclic heteroaryl ring having 1 to 3 heteroatoms). Non-limiting examples include indolyl, isoindolyl, benzothienyl, benzofuranyl, dibenzofuranyl, indazolyl, benzimidazolyl, benzothiazolyl, benzothiadiazolyl, benzoxazolyl, quinolyl, isoquinolyl, cinnolinyl, phthalazinyl, quinazolinyl, quinoxalinyl, 4H-quinolizinyl, carbazolyl, acridinyl, phenazinyl, phenothiazinyl, phenoxazinyl, tetrahydroquinolinyl, tetrahydroisoquinolinyl, pyrido[2,3-b]-1,4-oxazin-3(4H)-one, and benzisoxazolyl. The term "heteroaryl" may be used interchangeably with the terms "heteroaryl ring," "heteroaryl group," or "heteroaromatic," any of which terms include optionally substituted rings.

[0056] Heteroatom: The term "heteroatom" refers to one or more of oxygen, sulfur, nitrogen, phosphorus, or silicon (including any oxidized form of nitrogen, sulfur, phosphorus, or silicon; the quaternized form of any basic nitrogen; or a substitutable nitrogen of a heterocyclic ring, such as N (as in 3,4-dihydro-2H-pyrrolyl), NH (as in pyrrolidinyl), or NR + (as in N-substituted pyrrolidinyl).

[0057] Heterocycle: The terms "heterocycle", "heterocyclyl", "heterocyclic radical", and "heterocyclic ring" are used interchangeably herein and refer to a stable 3-8 membered monocyclic, 7-12 membered bicyclic, or 10-16 membered polycyclic heterocyclic moiety that is either saturated or partially unsaturated and has, in addition to carbon atoms, one or more heteroatoms, such as 1-4, as defined above. When used in reference to a ring atom of a heterocycle, the term "nitrogen" includes substituted nitrogen. As an example, in a saturated or partially unsaturated ring having 0-3 heteroatoms selected from oxygen, sulfur, or nitrogen, the nitrogen can be N (as in 3,4-dihydro-2H-pyrrolyl), NH (as in pyrrolidinyl), or NR + (as in N-substituted pyrrolidinyl). The heterocyclic ring can be attached to its pendant group at any heteroatom or carbon atom that results in a stable structure, and any of the ring atoms can be optionally substituted. Examples of such saturated or partially unsaturated heterocyclic radicals include, but are not limited to, azetidinyl, oxetanyl, tetrahydrofuranyl, tetrahydrothienyl, pyrrolidinyl, piperidinyl, decahydroquinolinyl, oxazolidinyl, piperazinyl, tetrahydropyranyl, dioxanyl, dioxolanyl, diazepinyl, oxazepinyl, thiazepinyl, morpholinyl, thiamorpholinyl, and [ka] Heterocyclyl groups can be monocyclic, bicyclic, tricyclic, or polycyclic, preferably monocyclic, bicyclic, or tricyclic, more preferably monocyclic or bicyclic. Bicyclic heterocyclic rings also include groups in which a heterocyclic ring is fused to one or more aryl rings. Exemplary bicyclic heterocyclic groups include indolinyl, isoindolinyl, benzodioxolyl, 1,3-dihydroisobenzofuranyl, 2,3-dihydrobenzofuranyl, and tetrahydroquinolinyl. Bicyclic heterocyclic rings can also be spirocyclic ring systems (e.g., 7-11 membered spirocyclic fused heterocyclic rings having, in addition to carbon atoms, one or more heteroatoms (e.g., 1, 2, 3, or 4 heteroatoms) as defined above). Bicyclic heterocyclic rings can also be bridged ring systems (eg, 7-11 membered bridged heterocyclic rings having 1, 2, or 3 bridging atoms).

[0058] Inhibitor: As used herein, the term "inhibitor" refers to an entity, condition, or event whose presence, level, or extent correlates with a decrease in the level or activity of a target. In some embodiments, an inhibitor may act directly (where it directly affects a target, e.g., by binding to the target), and in some embodiments, an inhibitor may act indirectly (where it affects the target by interacting with and / or altering a regulator of the target such that the level and / or activity of the target is reduced). In some embodiments, an inhibitor is one whose presence or level correlates with a target level or activity that is reduced compared to a particular reference level or activity (e.g., observed under appropriate reference conditions, such as the presence of a known inhibitor or the absence of the inhibitor in question).

[0059] In vitro: As used herein, the term "in vitro" refers to events that take place not within a multicellular organism but in an artificial environment, e.g., in a test tube or reaction vessel, in cell culture, etc.

[0060] Isolated: As used herein, refers to substances and / or entities that are (1) separated from at least some of the components with which they are initially produced (in nature and / or in an experimental setting) and / or (2) designed, produced, prepared, and / or manufactured by the hand of man. Isolated substances and / or entities can be separated from about 10%, about 20%, about 30%, about 40%, about 50%, about 60%, about 70%, about 80%, 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% of the other components with which they are initially produced. In some embodiments, an 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 more than about 99% pure. As used herein, a substance is "pure" if it is substantially free of other components. In some embodiments, as will be understood by those skilled in the art, a substance may still be considered "isolated" or even "pure" after being combined with certain other components, such as, for example, one or more carriers or excipients (e.g., buffers, solvents, water, etc.), and in such embodiments, the percentage of isolation or purity of the substance is calculated without including such carriers or excipients. By way of example, in some embodiments, a biological polymer, such as a polypeptide or polynucleotide, occurring in nature is considered to be "isolated" if: a) its origin or source is free from association with some or all of the components that accompany it in its natural state in nature; b) it is substantially free of other polypeptides or nucleic acids of the same species from the species that produces it in nature; or c) it is expressed by or associated with components from a cell or other expression system that is not of the species that produces it in nature. Thus, for example, in some embodiments, a polypeptide that is chemically synthesized or synthesized in a cellular system other than the one that produces it in nature is considered to be an "isolated" polypeptide.Alternatively, or in addition, in some embodiments, a polypeptide that has been subjected to one or more purification techniques may be considered to be an "isolated" polypeptide to the extent that it has been separated from a) other components with which it is associated in nature, and / or b) other components with which it was associated when originally produced.

[0061] In vivo: As used herein, refers to events that occur within multicellular organisms, such as humans and non-human animals. In the context of cell-based systems, the term can be used to refer to events that occur within living cells (as opposed to, for example, in vitro systems).

[0062] Linker: As used herein, it is used to refer to that portion of a multi-element agent that connects different elements to one another. For example, one of skill in the art will understand that a polypeptide that includes two or more functional or organizational domains in its structure will often include a stretch of amino acids between such domains that connects them to one another. In some embodiments, a polypeptide that includes a linker element has an overall structure of the general form S1-L-S2, where S1 and S2 may represent two domains that may be the same or different and are related to one another by the linker. In some embodiments, the polypeptide linker is at least 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 35, 40, 45, 50, 55, 60, 65, 70, 75, 80, 85, 90, 95, 100 or more amino acids in length. In some embodiments, the linker is characterized by not tending to adopt a rigid three-dimensional structure, but rather providing flexibility to the polypeptide.A variety of different linker elements can be appropriately used when engineering polypeptides (e.g., fusion polypeptides) known in the art (e.g., Holliger, P., et al. (1993) Proc. Natl. Acad. Sci. USA 90:6444-6448; Poljak, RJ, et al. (1994) Structure 2:1 121-1123).

[0063] Nanoparticles: As used herein, the term "nanoparticles" refers to particles having a diameter of less than 1000 nanometers (nm). In some embodiments, nanoparticles have a diameter of less than 300 nm as defined by the National Science Foundation. In some embodiments, nanoparticles have a diameter of less than 100 nm as defined by the National Institutes of Health. In some embodiments, nanoparticles are micelles in that they contain an enclosed compartment separated from the bulk solution by a micellar membrane, typically composed of amphiphilic entities that surround and encapsulate a space or compartment (e.g., to define a lumen). In some embodiments, the micellar membrane is composed of at least one polymer, such as, for example, a biocompatible and / or biodegradable polymer. In some embodiments, the lipid nanoparticles described herein may have an average hydrodynamic diameter of about 30 to about 170 nm. In some embodiments, the lipid nanoparticles described herein may have an average hydrodynamic diameter of about 30 nm, 35 nm, 40 nm, 45 nm, 50 nm, 55 nm, 60 nm, 65 nm, 70 nm, 75 nm, 80 nm, 85 nm, 90 nm, 95 nm, 100 nm, 105 nm, 110 nm, 115 nm, 120 nm, 125 nm, 130 nm, 135 nm, 140 nm, 145 nm, 150 nm, 155 nm, 160 nm, 165 nm, 170 nm, or any range having endpoints defined by any two of the foregoing values. For example, in some embodiments, the lipid nanoparticles described herein have an average hydrodynamic diameter of 50 nm to 100 nm.

[0064] Nanoparticle composition: As used herein, the term "nanoparticle composition" refers to a composition containing at least one nanoparticle and at least one additional agent or component. In some embodiments, the nanoparticle composition contains a population of substantially uniform nanoparticles as described herein.

[0065] Nucleic acid: as used herein in its broadest sense refers to any compound and / or substance that is or can be incorporated into an oligonucleotide chain. In some embodiments, a nucleic acid is a compound and / or substance that is or can be incorporated into an oligonucleotide chain via a phosphodiester bond. As will be clear from the context, in some embodiments, a "nucleic acid" refers to an individual nucleic acid residue (e.g., a nucleotide and / or a nucleoside), and in some embodiments, a "nucleic acid" refers to an oligonucleotide chain that includes individual nucleic acid residues. In some embodiments, a "nucleic acid" is or includes RNA, and in some embodiments, a "nucleic acid" is or includes DNA. In some embodiments, a nucleic acid is, includes, or consists of one or more naturally occurring nucleic acid residues. In some embodiments, a nucleic acid is, includes, or consists of one or more nucleic acid analogs. In some embodiments, a nucleic acid analog differs from a nucleic acid in that it does not utilize a phosphodiester backbone. For example, in some embodiments, the nucleic acid is, comprises, or consists of one or more "peptide nucleic acids," which are known in the art and have peptide bonds in the backbone instead of phosphodiester bonds, and are considered within the scope of the present invention. Alternatively or additionally, in some embodiments, the nucleic acid has one or more phosphorothioate and / or 5'-N-phosphoramidite linkages rather than phosphodiester linkages. In some embodiments, the nucleic acid is, comprises, or consists of one or more naturally occurring nucleosides (e.g., adenosine, thymidine, guanosine, cytidine, uridine, deoxyadenosine, deoxythymidine, deoxyguanosine, and deoxycytidine).In some embodiments, the nucleic acid is, comprises, or consists of one or more nucleoside analogs (e.g., 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, methylated bases, intercalated bases, and combinations thereof). In some embodiments, the nucleic acid comprises one or more modified sugars (e.g., 2'-fluororibose, ribose, 2'-deoxyribose, arabinose, and hexose) compared to the sugars in naturally occurring nucleic acids. In some embodiments, the nucleic acid has a nucleotide sequence that encodes a functional gene product, such as RNA or a protein. In some embodiments, the nucleic acid comprises one or more introns. In some embodiments, the nucleic acid is prepared by one or more of isolation from a natural source, enzymatic synthesis by polymerization based on a complementary template (in vivo or in vitro), reproduction in a recombinant cell or system, and chemical synthesis. In some embodiments, the nucleic acid is at least 3, 4, 5, 6, 7, 8, 9, 10, 15, 20, 25, 30, 35, 40, 45, 50, 55, 60, 65, 70, 75, 80, 85, 90, 95, 100, 110, 120, 130, 140, 150, 160, 170, 180, 190, 20, 225, 250, 275, 300, 325, 350, 375, 400, 425, 450, 475, 500, 600, 700, 800, 900, 1000, 1500, 2000, 2500, 3000, 3500, 4000, 4500, 5000 or more residues in length. In some embodiments, the nucleic acid is partially or completely single-stranded, in some embodiments, the nucleic acid is partially or completely double-stranded, in some embodiments, the nucleic acid has a nucleotide sequence that includes at least one element that encodes a polypeptide or is the complement of a sequence that encodes a polypeptide.In some embodiments, the nucleic acid has enzymatic activity.

[0066] Operably linked: As used herein, refers to a juxtaposition in which the described components are in a relationship permitting them to function in their intended manner. A control element "operably linked" to a functional element is associated with the functional element in a manner such that expression and / or activity of the functional element is achieved under conditions compatible with the control element. In some embodiments, an "operably linked" control element is contiguous (e.g., covalently linked) with a coding element of interest, and in some embodiments, the control element acts in trans or on the functional element of interest.

[0067] Parenteral: As used herein, the phrases "parenteral administration" and "administered parenterally" have their art-understood meaning referring to modes of administration other than enteral and topical administration, usually by injection, including, but not limited to, intravenous, intramuscular, intraarterial, intrathecal, intracapsular, intraorbital, intraventricular, intradermal, intraperitoneal, transtracheal, subcutaneous, subcuticular, intraarticular, subcapsular, subarachnoid, intraspinal, and intrasternal injection and infusion.

[0068] Patient: As used herein, the term "patient" refers to any organism to which a provided composition is or can be administered, for example, experimental, diagnostic, preventative, cosmetic, and / or therapeutic purposes. Exemplary patients include animals (e.g., mammals such as mice, rats, rabbits, non-human primates, and / or humans). In some embodiments, the patient is human. In some embodiments, the patient is suffering from or susceptible to one or more disorders or conditions. In some embodiments, the patient exhibits one or more symptoms of a disorder or condition. In some embodiments, the patient has been diagnosed as having one or more disorders or conditions. In some embodiments, the disorder or condition is or includes cancer, or one or more tumors. In some embodiments, the patient is undergoing or has undergone a certain therapy to diagnose and / or treat a disease, disorder, or condition.

[0069] Pharmaceutical composition: As used herein, the term "pharmaceutical composition" refers to an active agent formulated together with one or more pharma- ceutically acceptable carriers. In some embodiments, the active agent is present in an amount of unit doses suitable for administration in a treatment regimen that exhibits a statistically significant probability of achieving a predetermined therapeutic effect when administered to a relevant population. In some embodiments, the pharmaceutical composition may be specially formulated for administration in solid or liquid form, including oral administration, e.g., drenches (aqueous or non-aqueous solutions or suspensions), tablets, e.g., buccal, sublingual, and those targeted for systemic absorption, boluses, powders, granules, pastes for application to the tongue; parenteral administration, e.g., as a sterile solution or suspension, or sustained release formulation, e.g., subcutaneous, intramuscular, intravenous, or epidural injection; topical application, e.g., as a cream, ointment, or controlled release patch, or spray applied to the skin, lungs, or buccal cavity; vaginally or rectally, e.g., as a pessary, cream, or foam; sublingually; ophthalmic; transdermal; or compatible with nasal, pulmonary, and other mucosal surfaces.

[0070] Pharmaceutically acceptable: As used herein, the phrase "pharmacologically acceptable" refers to those compounds, materials, compositions, and / or dosage forms that are, within the scope of sound medical judgment, suitable for use in contact with the tissues of human beings and animals without undue toxicity, irritation, allergic response, or other problem or complication, commensurate with a reasonable benefit / risk ratio.

[0071] Pharmaceutically acceptable carrier: As used herein, the term "pharmaceutical acceptable carrier" refers to a pharma- ceutically acceptable material, composition, or vehicle, such as a material encapsulating a liquid or solid filler, diluent, excipient, or solvent, involved in carrying or transporting a compound of interest from one organ or part of the body to another organ or part of the body. Each carrier must be "acceptable" in the sense of being compatible with the other ingredients of the formulation and not harmful to the patient. Some examples of materials which can function as pharma- ceutically acceptable carriers include sugars such as lactose, glucose, and sucrose; starches such as corn starch and potato starch; cellulose and its derivatives such as sodium carboxymethylcellulose, ethylcellulose, and cellulose acetate; powdered tragacanth; malt; gelatin; talc; excipients such as cocoa butter and suppository waxes; oils such as peanut oil, cottonseed oil, safflower oil, sesame oil, olive oil, corn oil, and soybean oil; glycols such as propylene glycol; polyols such as glycerin, sorbitol, mannitol, and polyethylene glycol; esters such as ethyl oleate and ethyl laurate; agar; buffers such as magnesium hydroxide and aluminum hydroxide; alginic acid; pyrogen-free water; isotonic saline; Ringer's solution; ethyl alcohol; pH buffer solutions; polyesters, polycarbonates, and / or polyanhydrides; and other non-toxic compatible substances used in pharmaceutical formulations.

[0072] Pharmaceutically acceptable salts: The term "pharmaceutical acceptable salts" as used herein refers to salts of such compounds that are suitable for use in a pharmaceutical context, i.e., salts that are suitable for use in contact with the tissues of humans and lower animals without undue toxicity, irritation, allergic response, etc., within the scope of sound medical judgment, and commensurate with a reasonable benefit / risk ratio. Pharmaceutically acceptable salts are well known in the art. For example, S. M. Berge, et al., provide a detailed description of pharmaceutical acceptable salts in J. Pharmaceutical Sciences, 66:1-19 (1977). In some embodiments, pharmaceutical acceptable salts include non-toxic acid addition salts, which are salts of amino groups formed with inorganic acids such as hydrochloric acid, hydrobromic acid, phosphoric acid, sulfuric acid, and perchloric acid, or with organic acids such as acetic acid, maleic acid, tartaric acid, citric acid, succinic acid, or malonic acid, or by using other methods used in the art, such as ion exchange. In some embodiments, pharma- ceutically acceptable salts include, but are not limited to, adipate, alginate, ascorbate, aspartate, benzenesulfonate, benzoate, bisulfate, borate, butyrate, camphorate, camphorsulfonate, citrate, cyclopentanepropionate, digluconate, dodecyl sulfate, ethanesulfonate, formate, fumarate, glucoheptonate, glycerophosphate, gluconate, hemisulfate, heptanoate, hexanoate, hydrogen iodide, 2-hydroxybenzoate, benzoic acid salt ... Examples of the salts include hydroxy-ethanesulfonate, lactobionate, lactate, laurate, lauryl sulfate, malate, maleate, malonate, methanesulfonate, 2-naphthalenesulfonate, nicotinate, nitrate, oleate, oxalate, palmitate, pamoate, pectinate, persulfate, 3-phenylpropionate, phosphate, picrate, pivalate, propionate, stearate, succinate, sulfate, tartrate, thiocyanate, p-toluenesulfonate, undecanoate, valerate, etc. Representative alkali or alkaline earth metal salts include sodium, lithium, potassium, calcium, magnesium, etc.In some embodiments, pharma- ceutically acceptable salts include non-toxic ammonium, quaternary ammonium, and amine cations formed, where appropriate, using counterions such as halides, hydroxides, carboxylates, sulfates, phosphates, nitrates, alkyls having 1 to 6 carbon atoms, sulfonates, and arylsulfonates.

[0073] Prevent or Prevention: As used herein, when used in relation to the occurrence of a disease, disorder, and / or condition, refers to reducing the risk of developing the disease, disorder, and / or condition and / or delaying the onset of one or more characteristics or symptoms of the disease, disorder, or condition. Prevention may be considered complete if it has delayed the onset of the disease, disorder, or condition for a predefined period of time.

[0074] Polypeptide: The term "polypeptide" as used herein generally has its art-recognized meaning of a polymer of at least three amino acids. Those of skill in the art will appreciate that the term "polypeptide" is intended to be general enough to encompass not only polypeptides having the complete sequences listed herein, but also polypeptides that represent functional fragments of such complete polypeptides (i.e., fragments that retain at least one activity). Moreover, those of skill in the art will appreciate that protein sequences generally tolerate some substitutions without destroying activity. Thus, any polypeptide that retains activity and shares one or more highly conserved regions with another polypeptide of the same genus, usually encompassing at least 3-4, and often up to 20 or more amino acids, including at least one region of very high identity, usually at least about 30-40% overall sequence identity, often more than about 50%, 60%, 70%, or 80%, and even usually often more than 90%, or even 95%, 96%, 97%, 98%, or 99%, is encompassed within the related term "polypeptide" as used herein. Polypeptides may contain L-amino acids, D-amino acids, or both, and may contain any of a variety of amino acid modifications or analogs known in the art. Useful modifications include, for example, terminal acetylation, amidation, methylation, and the like. In some embodiments, proteins may contain natural amino acids, unnatural amino acids, synthetic amino acids, and combinations thereof. The term "peptide" is generally used to refer to a polypeptide having a length of less than about 100 amino acids, less than about 50 amino acids, less than 20 amino acids, or less than 10 amino acids. In some embodiments, the protein is an antibody, an antibody fragment, a biologically active portion thereof, and / or a characteristic portion thereof.

[0075] Prevention: The term "prevention" as used herein refers to delaying the onset and / or reducing the frequency and / or severity of one or more symptoms of a particular disease, disorder, or condition. In some embodiments, prevention is assessed on a population basis, such that an agent is considered to "prevent" a particular disease, disorder, or condition if a statistically significant reduction in the onset, frequency, and / or intensity of one or more symptoms of the disease, disorder, or condition is observed in a population susceptible to the disease, disorder, or condition. Prevention can be considered complete if it delays the onset of the disease, disorder, or condition for a predefined period of time.

[0076] Protecting group: As used herein, the phrase "protecting group" refers to a temporary substituent that protects a potentially reactive functional group from undesired chemical transformations. Examples of such protecting groups include esters of carboxylic acids, silyl ethers of alcohols, and acetals and ketals of aldehydes and ketones, respectively. "Si-protecting group" is a protecting group that contains a Si atom, such as Si-trialkyl (e.g., trimethylsilyl, tributylsilyl, t-butyldimethylsilyl), Si-triaryl, Si-alkyl-diphenyl (e.g., t-butyldiphenylsilyl), or Si-aryl-dialkyl (e.g., Si-phenyldialkyl). Generally, Si-protecting groups are attached to an oxygen atom. The field of protecting group chemistry has been reviewed (Greene, TW; Wuts, PGM Protective Groups in Organic Synthesis, 2nd ed.; Wiley: New York, 1991). Such protecting groups (and related protected moieties) are described in detail below.

[0077] Hydroxyl groups that can be protected are well known in the art and are described in Protecting Groups in Organic Synthesis, T.W. Greene and P.G.M. Huts, 3rd Edition, incorporated herein by reference in its entirety. rdedition, John Wiley & Sons, 1999. Examples of suitable protected hydroxyl groups further include, but are not limited to, esters, carbonates, sulfonates, allyl ethers, ethers, silyl ethers, alkyl ethers, arylalkyl ethers, and alkoxyalkyl ethers. Examples of suitable esters include formates, acetates, propionates, pentanoates, crotonates, and benzoates. Specific examples of suitable esters include formates, benzoyl formates, chloroacetates, trifluoroacetates, methoxyacetates, triphenylmethoxyacetates, p-chlorophenoxyacetates, 3-phenylpropionates, 4-oxopentanoates, 4,4-(ethylenedithio)pentanoates, pivalates (trimethylacetates), crotonates, 4-methoxy-crotonates, benzoates, p-benzylbenzoates, and 2,4,6-trimethylbenzoates. Examples of suitable carbonates include 9-fluorenylmethyl, ethyl, 2,2,2-trichloroethyl, 2-(trimethylsilyl)ethyl, 2-(phenylsulfonyl)ethyl, vinyl, allyl, and p-nitrobenzyl carbonate. Examples of suitable silyl ethers include trimethylsilyl, triethylsilyl, t-butyldimethylsilyl, t-butyldiphenylsilyl, triisopropylsilyl ether, and other trialkylsilyl ethers. Examples of suitable alkyl ethers include methyl, benzyl, p-methoxybenzyl, 3,4-dimethoxybenzyl, trityl, t-butyl, and allyl ethers, or derivatives thereof. Alkoxyalkyl ethers include acetals such as methoxymethyl, methylthiomethyl, (2-methoxyethoxy)methyl, benzyloxymethyl, beta-(trimethylsilyl)ethoxymethyl, and tetrahydropyran-2-yl ether. Examples of suitable arylalkyl ethers include benzyl, p-methoxybenzyl (MPM), 3,4-dimethoxybenzyl, O-nitrobenzyl, p-nitrobenzyl, p-halobenzyl, 2,6-dichlorobenzyl, p-cyanobenzyl, 2- and 4-picolyl ethers.

[0078] Protected amines are well known in the art and include those detailed in Greene (1999). Suitable mono-protected amines further include, but are not limited to, aralkylamines, carbamates, allylamines, amides, and the like. Examples of suitable mono-protected amino moieties include t-butyloxycarbonylamino (-NHBOC), ethyloxycarbonylamino, methyloxycarbonylamino, trichloroethyloxycarbonylamino, allyloxycarbonylamino (-NHAlloc), benzyloxocarbonylamino (-NHCBZ), allylamino, benzylamino (-NHBn), fluorenylmethylcarbonyl (-NHFmoc), formamide, acetamide, chloroacetamide, dichloroacetamide, trichloroacetamide, phenylacetamide, trifluoroacetamide, benzamide, t-butyldiphenylsilyl, and the like. Suitable di-protected amines include amines substituted with two substituents independently selected from those listed above as mono-protected amines, and further include cyclic imides such as phthalimides, maleimides, succinimides, etc. Suitable di-protected amines also include pyrroles, such as 2,2,5,5-tetramethyl-[1,2,5]azadisilolidine, and azides.

[0079] Protected aldehydes are well known in the art and include those detailed in Greene (1999). Suitable protected aldehydes further include, but are not limited to, acyclic acetals, cyclic acetals, hydrazones, imines, and the like. Examples of such groups include dimethyl acetal, diethyl acetal, diisopropyl acetal, dibenzyl acetal, bis(2-nitrobenzyl) acetal, 1,3-dioxane, 1,3-dioxolane, semicarbazones, and derivatives thereof.

[0080] Protected carboxylic acids are well known in the art and include those detailed in Greene (1999). Suitable protected carboxylic acids include, but are not limited to, optionally substituted C 1-6 Further included are aliphatic esters, optionally substituted aryl esters, silyl esters, activated esters, amides, hydrazides, etc. Examples of such ester groups include methyl, ethyl, propyl, isopropyl, butyl, isobutyl, benzyl, and phenyl esters, each group being optionally substituted. Additional suitable protected carboxylic acids include oxazolines and orthoesters.

[0081] Protected thiols are well known in the art and include those detailed in Greene (1999).Suitably protected thiols further include, but are not limited to, disulfides, thioethers, silyl thioethers, thioesters, thiocarbonates, and thiocarbamates.Examples of such groups include, but are not limited to, alkyl thioethers, benzyl and substituted benzyl thioethers, triphenylmethyl thioethers, and trichloroethoxycarbonyl thioesters, to name just a few.

[0082] Protein: The term "protein" as used herein refers to one or more polypeptides that function as separate units. When a single polypeptide is a separate functional unit and does not require permanent or temporary physical association with other polypeptides to form a separate functional unit, the terms "polypeptide" and "protein" may be used interchangeably. When a separate functional unit is composed of more than one polypeptide that physically associates with each other, the term "protein" may be used to refer to multiple polypeptides that are physically associated and function together as a separate unit. In some embodiments, a protein may include moieties other than amino acids (e.g., may be a glycoprotein, proteoglycan, etc.) and / or may be processed or modified. One of skill in the art will understand that in some embodiments, the term "protein" may refer to the complete polypeptide chain produced by a cell (e.g., with or without a signal sequence) and / or a form that is active within the cell (e.g., a truncated or complexed form). In some embodiments where a protein is composed of multiple polypeptide chains, such chains may be covalently associated with each other, e.g., by one or more disulfide bonds, or may be associated by other means.

[0083] Pure: As used herein, an agent or entity is "pure" if it is substantially free of other components. For example, a preparation that contains greater than about 90% of a particular agent or entity is typically considered to be a pure preparation. In some embodiments, the agent or entity is at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% pure.

[0084] Reference: as used herein describes a standard or control against which a comparison is made. For example, in some embodiments, an agent, animal, individual, population, sample, sequence, or value of interest is compared to a reference or control agent, animal, individual, population, sample, sequence, or value. In some embodiments, the reference or control is tested and / or determined substantially simultaneously with the test or determination of interest. In some embodiments, the reference or control is a historical reference or control, optionally embodied in a tangible medium. Typically, as will be understood by those of skill in the art, a reference or control is determined or characterized under conditions or circumstances equivalent to those being evaluated. Those of skill in the art will understand when there is sufficient similarity to justify reliance and / or comparison on a particular possible reference or control.

[0085] Sample: As used herein, the term "sample" typically refers to an aliquot of material obtained or derived from a source of interest as described herein. In some embodiments, the source of interest is a biological or environmental source. In some embodiments, the source of interest may be or include a cell, or an organism such as a microorganism, a plant, or an animal (e.g., a human). In some embodiments, the source of interest is or includes a biological tissue or fluid. In some embodiments, the biological tissue or fluid may be or include amniotic fluid, aqueous humor, peritoneal fluid, bile, bone marrow, blood, breast milk, cerebrospinal fluid, earwax, chyle, chime, ejaculate, endolymph, exudate, feces, gastric acid, gastric juice, lymph, mucus, pericardial fluid, perilymph, peritoneal fluid, pleural fluid, pus, eye discharge, saliva, sebum, semen, serum, smegma, sputum, synovial fluid, sweat, tears, urine, vaginal secretions, vitreous, vomit, and / or combinations or components thereof. In some embodiments, the biological fluid may be or include intracellular fluid, extracellular fluid, intravascular fluid (plasma), interstitial fluid, lymph, and / or intercellular fluid. In some embodiments, the biological fluid may be or include phytoexudates. In some embodiments, the biological tissue or sample may be obtained, for example, by aspiration, biopsy (e.g., fine needle biopsy or tissue biopsy), swab (e.g., oral, nasal, skin, or vaginal swab), scraping, surgery, lavage or washing (e.g., bronchoalveolar, ductal, nasal, ocular, oral, uterine, vaginal, or other lavage or washing). In some embodiments, the biological sample is or comprises cells obtained from an individual. In some embodiments, the sample is a "primary sample" obtained directly from the source of interest by any suitable means. In some embodiments, as will be clear from the context, the term "sample" refers to a preparation obtained by processing the primary sample (e.g., by removing one or more components and / or adding one or more agents). For example, filtering using a semi-permeable membrane. Such a "processed sample" may comprise, for example, nucleic acids or proteins extracted from the sample or obtained by subjecting the primary sample to one or more techniques, such as amplification or reverse transcription of nucleic acids, isolation and / or purification of certain components.

[0086] Stable nanoparticle compositions: The term "stable" as applied to compositions herein means that the compositions maintain one or more aspects of their physical structure (e.g., particle size range and / or distribution) over a period of time. In some embodiments, a stable nanoparticle composition is one in which the mean particle size, maximum particle size, particle size range, and / or particle size distribution (i.e., percentage of particles above a specified size and / or outside a specified size range) are maintained under defined conditions for a period of time. In some embodiments, a stable provided composition is one in which biologically relevant activity is maintained for a period of time. In some embodiments, the period of time is at least about 1 hour, and in some embodiments, the period of time is about 5 hours, about 10 hours, about one (1) day, about one (1) week, about two (2) weeks, about one (1) month, about two (2) months, about three (3) months, about four (4) months, about five (5) months, about six (6) months, about eight (8) months, about ten (10) months, about twelve (12) months, about twenty-four (24) months, about thirty-six (36) months, or more. In some embodiments, the period of time is within a range of about one (1) day to about twenty-four (24) months, about two (2) weeks to about twelve (12) months, about two (2) months to about five (5) months, etc. For example, a nanoparticle composition is stable if a population of nanoparticles is subjected to storage, temperature changes, and / or pH changes for extended periods of time and the majority of the nanoparticles in the composition maintain diameters within the recited ranges. In some embodiments, the stable composition is stable at ambient conditions. In some embodiments, the stable composition is stable under biological conditions (i.e., in phosphate buffered saline at 37° C.).

[0087] Sterolyl: The term "sterolyl" as used herein refers to a 17-membered fused polycyclic ring moiety that is saturated or partially unsaturated, substituted with at least one hydroxyl group, and has a single point of attachment to the remainder of the molecule at any substitutable carbon or oxygen atom. In some embodiments, the sterolyl group is a cholesterolyl group, or a variant or derivative thereof. In some embodiments, the cholesterolyl group is modified. In some embodiments, the cholesterolyl group is an oxidized cholesterolyl group (e.g., oxidized at the beta ring structure or the hydrocarbon tail structure). In some embodiments, the cholesterolyl group is an esterified cholesterolyl group. In some embodiments, the cholesterolyl group is a phytosterolyl group. Exemplary sterolyl groups include, but are not limited to, 25-hydroxycholesterolyl (25-OH), 20α-hydroxycholesterolyl (20α-OH), 27-hydroxycholesterolyl, 6-keto-5α-hydroxycholesterolyl, 7-ketocholesterolyl, 7β-hydroxycholesterolyl, 7α-hydroxycholesterolyl, 7β-25-dihydroxycholesterolyl, beta-sitosterolyl, stigmasterolyl, brassicasterolyl, and campesterolyl.

[0088] Subject: As used herein, the term "subject" refers to an organism, typically a mammal (e.g., in some embodiments, a human, including prenatal human forms). In some embodiments, the subject suffers from a relevant disease, disorder, or condition. In some embodiments, the subject is predisposed to a disease, disorder, or condition. In some embodiments, the subject exhibits one or more symptoms or characteristics of a disease, disorder, or condition. In some embodiments, the subject does not exhibit any symptoms or characteristics of a disease, disorder, or condition. In some embodiments, the subject is one who has one or more characteristics that are predisposed to or at risk for being predisposed to a disease, disorder, or condition. In some embodiments, the subject is a patient. In some embodiments, the subject is an individual for whom and / or upon whom diagnosis and / or treatment is to be performed.

[0089] Substantially: As used herein, the term "substantially" refers to a qualitative state that indicates the total or nearly total extent or degree of a desired characteristic or property. Those skilled in the art of biology will understand that biological and chemical phenomena rarely, if ever, go completely to completion and / or toward complete completion, or achieve or avoid absolute results. Thus, the term "substantially" is used herein to capture the potential lack of completeness inherent in many biological and chemical phenomena.

[0090] Substituted or Optionally Substituted: The compounds of the present disclosure described herein may contain moieties that are optionally substituted and / or substituted. In general, the term "substituted," whether preceded by the term "optionally," means that one or more hydrogens of the specified moiety are replaced with a suitable substituent. "Substituted" applies to one or more hydrogens, either explicitly or implicitly from the structure (e.g., [ka] At least [ka] Refers to, [ka] At least [ka] (refers to "optionally substituted"). Unless otherwise indicated, an "optionally substituted" group may have a suitable substituent at each substitutable position of the group, and when more than one position in any given structure may be substituted with more than one substituent selected from a specified group, the substituents may be the same or different at all positions. The combinations of substituents envisioned by the present disclosure are preferably those that result in the formation of stable or chemically feasible compounds. The term "stable", as used herein, refers to compounds that are substantially unchanged when subjected to conditions that permit their production, detection, and, in certain embodiments, their recovery, purification, and use for one or more of the purposes disclosed herein. Groups described as "substituted" preferably have 1-6 substituents, more preferably 1-2 substituents. Groups described as "optionally substituted" may be unsubstituted or "substituted" as described above.

[0091] Suitable monovalent substituents include halogen; -(CH 2 ) 0-4 R°;-(CH 2 ) 0-4 OR°;-O(CH 2 ) 0-4 R°, -O-(CH 2 ) 0-4 C(O)OR°;-(CH 2 ) 0-4 CH(OR°) 2 ;R° may be substituted -(CH 2 ) 0-4 Ph; R° may be substituted with -(CH 2 ) 0-4 O(CH 2 ) 0-1 Ph; may be substituted with R° -CH=CHPh; may be substituted with R° -(CH 2 ) 0-4 O(CH 2 ) 0-1 -Pyridinyl; -NO 2 ;-CN;-N 3 ;-(CH 2 ) 0-4 N(R°) 2 ;-(CH 2 ) 0-4N(R°)C(O)R°;-N(R°)C(S)R°;-(CH 2 ) 0-4 N(R°)C(O)NR° 2 ;-N(R°)C(S)NR° 2 ;-(CH 2 ) 0-4 N(R°)C(O)OR°;-N(R°)N(R°)C(O)R°;-N(R°)N(R°)C(O)NR° 2 ;-N(R°)N(R°)C(O)OR°;-(CH 2 ) 0-4 C(O)R°;-C(S)R°;-(CH 2 ) 0-4 C(O)OR°;-(CH 2 ) 0-4 C(O)SR°;-(CH 2 ) 0-4 C(O)OSiR° 3 ;-(CH 2 ) 0-4 OC(O)R°;-OC(O)(CH 2 ) 0-4 SR°-、-SC(S)SR°;-(CH 2 ) 0-4 SC(O)R°;-(CH 2 ) 0-4 C(O)NR° 2 ;-C(S)NR° 2 ;-C(S)SR°;-(CH 2 ) 0-4 OC(O)NR° 2 ;-C(O)N(OR°)R°;-C(O)C(O)R°;-C(O)CH 2 C(O)R°;-C(NOR°)R°;-(CH 2 ) 0-4 SSR°;-(CH 2 ) 0-4 S(O) 2 R°;-(CH 2 ) 0-4 S(O) 2 OR°;-(CH 2 ) 0-4 OS(O) 2 R°;-S(O) 2 NR° 2 ;-(CH 2 ) 0-4S(O)R°;-N(R°)S(O) 2 NR° 2 ;-N(R°)S(O) 2 R°;-N(OR°)R°;-C(NH)NR° 2 ;-P(O) 2 R°;-P(O)R° 2 ;-OP(O)R° 2 ;-OP(O)(OR°) 2 ;-SiR° 3 ;-OSiR° 3 ;-(C 1-4 Linear or branched alkylene)ON(R°) 2 ;or-(C 1-4 Linear or branched alkylene)C(O)ON(R°) 2 wherein each R° can be substituted as defined below and is independently hydrogen, C 1-6 Aliphatic, -CH 2 Ph, -O(CH 2 ) 0-1 Ph, -CH 2 -(5-6 membered heteroaryl ring), or a 5-6 membered saturated, partially unsaturated ring having 0-4 heteroatoms independently selected from nitrogen, oxygen, or sulfur, or an aryl ring, as defined above, but taken together with the intervening atoms, two independent occurrences of R° form a 3-12 membered saturated, partially unsaturated ring having 0-4 heteroatoms independently selected from nitrogen, oxygen, or sulfur, or an aryl monocyclic or bicyclic ring, which may be substituted as defined below.

[0092] Suitable monovalent substituents on R° (or the ring formed, together with the intervening atoms, by two independent occurrences of R°) are independently halogen, -(CH 2 ) 0-2 R ● , -(Halo R ● ), -(CH 2 ) 0-2 OH, -(CH 2 ) 0-2 OR ● , -(CH 2 ) 0-2 CH(OR ● )2 ;-O(Halo R ● ), -CN, -N 3 , -(CH 2 ) 0-2 C(O)R ● , -(CH 2 ) 0-2 C(O)OH, -(CH 2 ) 0-2 C(O)OR ● , -(CH 2 ) 0-2 S.R. ● , -(CH 2 ) 0-2 SH, -(CH 2 ) 0-2 NH 2 , -(CH 2 ) 0-2 NHR ● , -(CH 2 ) 0-2 NR ● 2 , -NO 2 , -SiR ● 3 , -OSiR ● 3 , -C(O)SR ● 、 -(C 1-4 Linear or branched alkylene)C(O)OR ● , or -SSR ● And each R ● is unsubstituted or, if preceded by "halo", is substituted only with one or more halogens, and C 1-4 Aliphatic, -CH 2 Ph, -O(CH 2 ) 0-1 Ph or a 5-6 membered saturated, partially unsaturated ring having 0-4 heteroatoms independently selected from nitrogen, oxygen, or sulfur, or an aryl ring. Suitable divalent substituents on a saturated carbon atom of R° include =O and =S.

[0093] Suitable divalent substituents include the following: =O, =S, =NNR * 2 , =NNHC(O)R * , =NNHC(O)OR *, =NNHS(O) 2 R * , =NR * , =NOR * , -O(C(R * 2 )) 2-3 O- or -S(C(R * 2 )) 2-3 S-, each of which is an independent R * occurrences of 1-6 Aliphatic or unsubstituted 5-6 membered saturated, partially unsaturated, or aryl ring having 0-4 heteroatoms independently selected from nitrogen, oxygen, or sulfur. Suitable divalent substituents attached to the proximal substitutable carbon of the "optionally substituted" group include -O(CR * 2 ) 2-3 O-, and each independently R * occurrences of 1-6 It is selected from an aliphatic, or unsubstituted 5-6 membered saturated, partially unsaturated ring having 0-4 heteroatoms independently selected from nitrogen, oxygen, or sulfur, or an aryl ring.

[0094] R * Suitable substituents on the aliphatic group include halogen, -R ● , -(Halo R ● ), -OH, -OR ● , -O(HaloR ● ), -CN, -C(O)OH, -C(O)OR ● , -NH 2 , -NHR ● , -NR ● 2 , or -NO 2 Each R ● is unsubstituted or, if preceded by "halo", is substituted only with one or more halogens, and independently represents C 1-4 Aliphatic, -CH 2 Ph, -O(CH 2 ) 0-1Ph, or a 5-6 membered saturated, partially unsaturated ring having 0-4 heteroatoms independently selected from nitrogen, oxygen, or sulfur, or an aryl ring.

[0095] In some embodiments, suitable substituents on a substitutable nitrogen include -R † , -NR † 2 , -C(O)R † , -C(O)OR † , -C(O)C(O)R † , -C(O)CH 2 C(O)R † , -S(O) 2 R † , -S(O) 2 NR † 2 , -C(S)NR † 2 , -C(NH)NR † 2 , or -N(R † )S(O) 2 R † Each R † are independently hydrogen, C which may be substituted as defined below 1-6 an aliphatic, unsubstituted -OPh, or an unsubstituted 5-6 membered saturated, partially unsaturated, or aryl ring having 0-4 heteroatoms independently selected from nitrogen, oxygen, or sulfur, or as defined above but which, together with the intervening atoms, form two independent R † The occurrence of forms an unsubstituted 3-12 membered saturated, partially unsaturated ring having 0-4 heteroatoms independently selected from nitrogen, oxygen, or sulfur, or an aryl monocyclic or bicyclic ring.

[0096] R † Suitable substituents on the aliphatic groups are independently halogen, -R ● , -(Halo R ● ), -OH, -OR ● , -O(HaloR ● ), -CN, -C(O)OH, -C(O)OR ● , -NH 2 , -NHR● , -NR ● 2 , or -NO 2 And each R ● is unsubstituted or, if preceded by "halo", is substituted only with one or more halogens, and independently represents C 1-4 Aliphatic, -CH 2 Ph, -O(CH 2 ) 0-1 Ph, or a 5-6 membered saturated, partially unsaturated ring having 0-4 heteroatoms independently selected from nitrogen, oxygen, or sulfur, or an aryl ring.

[0097] Susceptible: An individual who is "susceptible" to a disease, disorder, or condition is at risk of developing the disease, disorder, or condition. In some embodiments, an individual who is susceptible to a disease, disorder, or condition does not exhibit any symptoms of the disease, disorder, or condition. In some embodiments, an individual who is susceptible to a disease, disorder, or condition has never been diagnosed with the disease, disorder, and / or condition. In some embodiments, an individual who is susceptible to a disease, disorder, or condition is an individual who has been exposed to conditions associated with the development of the disease, disorder, or condition. In some embodiments, the risk of developing a disease, disorder, and / or condition is a population-based risk (e.g., family members of an individual who suffers from the disease, disorder, or condition).

[0098] Systemic: As used herein, the phrases "systemic administration," "administered systemically," "peripheral administration," and "administered peripherally" have their art-recognized meanings referring to the administration of a compound or composition so that it enters the system of the recipient.

[0099] Tautomeric forms: As used herein, the phrase "tautomeric forms" is used to describe different isomeric forms of an organic compound that are capable of easy interconversion. Tautomers may be characterized by a topological shift of a hydrogen atom or a proton, accompanied by a switch of a single bond and an adjacent double bond. In some embodiments, tautomers may result from prototropic tautomerism (i.e., rearrangement of a proton). In some embodiments, tautomers may result from valence tautomerism (i.e., rapid rearrangement of bond electrons). All such tautomeric forms are intended to be included within the scope of the present disclosure. In some embodiments, tautomeric forms of a compound exist in transfer equilibrium with one another, such that attempts to prepare separate substances would result in the formation of a mixture. In some embodiments, tautomeric forms of a compound are separable and isolatable compounds. In some embodiments of the present disclosure, chemical compositions may be provided that are or include pure preparations of a single tautomeric form of a compound. In some embodiments, chemical compositions may be provided as mixtures of two or more tautomeric forms of a compound. In certain embodiments, such mixtures contain equal amounts of different tautomeric forms, and in certain embodiments, such mixtures contain unequal amounts of at least two different tautomeric forms of a compound. In some embodiments of the present disclosure, the chemical composition may contain all tautomeric forms of a compound. In some embodiments of the present disclosure, the chemical composition may contain fewer than all tautomeric forms of a compound. In some embodiments of the present disclosure, the chemical composition may contain one or more tautomeric forms of a compound in amounts that vary over time as a result of interconversion. In some embodiments of the present disclosure, the tautomer is keto-enol tautomerism. One of skill in the art will recognize that keto-enol tautomers can be "captured" (i.e., chemically modified to remain in the "enol" form) using any suitable reagent known in the chemical art to provide an enol derivative that can then be isolated using one or more suitable techniques known in the art. Unless otherwise indicated, the present disclosure encompasses all tautomeric forms of the relevant compounds, whether in pure form or mixed with each other.

[0100] Therapeutic Agent: As used herein, the phrase "therapeutic agent" refers to an agent that has a therapeutic effect and / or induces a desired biological and / or pharmacological effect when administered to a subject. In some embodiments, a therapeutic agent is any substance that can be used to alleviate, ameliorate, reduce, inhibit, prevent, delay the onset of, reduce the severity of, and / or reduce the incidence of one or more symptoms or characteristics of a disease, disorder, and / or condition.

[0101] Therapeutically effective amount: As used herein, the term "therapeutically effective amount" refers to an amount of a substance (e.g., a therapeutic agent, composition, and / or formulation) that induces a desired biological response when administered as part of a treatment regimen. In some embodiments, a therapeutically effective amount of a substance is an amount that, when administered to a subject suffering from or susceptible to a disease, disorder, and / or condition, is sufficient to treat, diagnose, inhibit, ameliorate, prevent, and / or delay the onset of the disease, disorder, and / or condition. As will be understood by one of skill in the art, the effective amount of a substance can vary depending on such factors as the desired biological endpoint, the substance delivered, the target cell or tissue, and the like. For example, an effective amount of a compound in a formulation for treating a disease, disorder, and / or condition is an amount that alleviates, ameliorates, relieves, inhibits, prevents, delays the onset of, reduces the severity of, and / or reduces the incidence of one or more symptoms or characteristics of the disease, disorder, and / or condition. In some embodiments, a therapeutically effective amount is administered in a single dose, and in some embodiments, multiple unit doses are required to deliver a therapeutically effective amount. The exact dosage will vary depending on a variety of factors, such as subject-specific variables (eg, age, health of the immune system, etc.), the disease, and the treatment being administered.

[0102] "Tissue" and / or "Organ": As used herein, the term "tissue" and / or "organ" refers to viable cellular material in aggregate form, e.g., small portions of organs, and dispersed cells, e.g., cells dispersed, isolated, and / or grown from muscle, cardiac muscle, liver, or kidney, including bone marrow cells and progeny cells, blood-producing stem cells and progeny, and various other blood elements, unless otherwise specified. In some embodiments, tissue and / or organ refers to kidney, heart liver, stomach, spleen, pancreas, lung, brain, eye, intestine, bladder, skin or skin tissue, blood vessels, veins, arteries, heart valves, sperm, and oocytes. As used herein, the term "organ" encompasses both solid organs, e.g., kidney, heart, liver, lung, and functional portions of organs, e.g., portions of skin, arteries, veins, transplantable liver lobes, kidneys, lung sections, etc.

[0103] Treatment: As used herein, the term "treatment" (also "treat" or "treating") refers to the administration of a therapy that partially or completely alleviates, improves, relieves, inhibits, delays the onset, reduces the severity, and / or reduces the incidence of one or more symptoms, characteristics, and / or causes of a particular disease, disorder, and / or condition. In some embodiments, such treatment may be of a subject who is not showing signs of the associated disease, disorder, and / or condition and / or of a subject who is showing only early signs of the disease, disorder, and / or condition. Alternatively or additionally, such treatment may be of a subject who shows one or more established signs of the associated disease, disorder, and / or condition. In some embodiments, the treatment may be of a subject who has been diagnosed as suffering from the associated disease, disorder, and / or condition. In some embodiments, the treatment may be of a subject who is known to have one or more susceptibility factors that are statistically correlated with an increased risk of developing the associated disease, disorder, and / or condition. Thus, in some embodiments, the treatment may be prophylactic and in some embodiments, therapeutic. DETAILED DESCRIPTION OF THE PREFERRED EMBODIMENTS

[0104] The present disclosure describes how the selection and combination of one or more of the components described herein influences the functional activity of lipid nanoparticles, such as desired tropism, stabilization, and drug delivery efficacy. In particular, the present invention provides compositions, preparations, nanoparticles, and / or nanomaterials for the delivery of therapeutic and / or prophylactic agents to target cells and / or tissues. For example, the present disclosure describes lipid compounds for use in the compositions, preparations, nanoparticles, and / or nanomaterials. In some embodiments, the compositions, preparations, nanoparticles, and / or nanomaterials include LNPs that carry cargo to designated target cells, tissues, and / or organs.

[0105] I. Lipid Nanoparticles (LNPs) The present invention provides compositions, preparations, and / or nanomaterials comprising lipid nanoparticles. In some embodiments, the lipid nanoparticles comprise one or more components. In some embodiments, the lipid nanoparticles comprise one or more components, such as a compound, an ionizable lipid, a sterol, a conjugate-linker lipid, and a phospholipid. In particular, the present disclosure describes that the selection and combination of one or more of the components described herein affects the characteristics of lipid nanoparticles, such as diameter, pKa, stabilization, and ionizability.

[0106] In particular, the present disclosure describes that the selection and combination of one or more of the components described herein affects the functional activity of lipid nanoparticles, such as tropism, stabilization, and drug delivery effectiveness.For example, the present disclosure describes that the combination of components may be better suited for the delivery of siRNA.As another example, the present disclosure describes that the combination of components may be better suited for the delivery of mRNA.As another example, the present disclosure describes that the combination of components may be better suited for the delivery of DNA.

[0107] In some embodiments, the lipid nanoparticles comprise one or more compounds as described herein. In some embodiments, the lipid nanoparticles comprise one or more ionizable lipids as described herein. In some embodiments, the lipid nanoparticles comprise one or more sterols as described herein. In some embodiments, the lipid nanoparticles comprise one or more conjugate-linker lipids as described herein. In some embodiments, the lipid nanoparticles comprise one or more phospholipids as described herein.

[0108] In some embodiments, the present disclosure provides ionizable lipids characterized by a core moiety that is substantially free of or completely devoid of stereocenters (e.g., benzene core moieties). Without wishing to be bound by any theory, the present disclosure provides insight that substantially pure stereoisomers of ionizable lipids containing certain non-planar core moieties that contain one or more stereocenters may be particularly difficult to prepare via asymmetric synthesis. Chiral separation may be available for bench-scale synthesis, but may not be suitable for industrial synthesis (e.g., scale-up synthesis). One advantage of certain ionizable lipids provided in the present disclosure is that this concern is obviated through the incorporation of a core moiety that is substantially free of or completely devoid of stereocenters (e.g., benzene core moieties). Thus, among other things, certain provided lipids may be particularly suitable for large-scale manufacturing and may provide certain advantages in these contexts.

[0109] In some embodiments, the present disclosure provides ionizable lipids characterized by a planar core moiety (e.g., a benzene core moiety). Without wishing to be bound by any theory, such planar features may affect the rigidity of the lipid bilayer structure within the corresponding compositions, preparations, nanoparticles, and / or nanomaterials, which would likely affect the encapsulation efficiency, diameter, and / or fusogenicity of the corresponding compositions, preparations, nanoparticles, and / or nanomaterials.

[0110] A. Compound Among other things, the present disclosure describes compositions, preparations, nanoparticles, and / or nanomaterials that include one or more compounds described herein.

[0111] In some embodiments, the present disclosure provides a compound of formula I: [ka] or a pharma- ceutically acceptable salt thereof, wherein: Each L 1 and L 1’ is independently -C(O)- or -C(O)O-; Each L 2 and L 2’ are independently a covalent bond, an optionally substituted divalent saturated or unsaturated linear or branched C 1 -C 12 a hydrocarbon chain, or [ka] and Each Cy A is independently an optionally substituted ring selected from phenylene, and a 3- to 7-membered saturated or partially unsaturated carbocyclene; each m is independently 0, 1, or 2; Each L 3 and L 3’ are independently a covalent bond, —C(O)O—, —OC(O)—, —O—, or —OC(O)O—; Each R 1 and R 1’ is independently an optionally substituted group, in which 1 to 3 methylene units are optionally and independently replaced by -O- or -NR-, and 1 -C 20 Hydrocarbon chains; 3- to 12-membered saturated or partially unsaturated carbocyclic rings; 1-adamantyl; 2-adamantyl; sterolyl; phenyl; and [ka] is an optionally substituted group selected from Each L 4 independently, divalent saturated or unsaturated linear or branched C 1 -C 6 is a hydrocarbon chain, Each A 1 and A 2 is independently an optionally substituted C 1 -C 20 Aliphatic or -L 5 -R 5 or Or A 1 and A 2 may be taken together with their intervening atoms to form an optionally substituted ring, [ka] During the ceremony, x is selected from 1 or 2; # is L 4 represents the attachment point to Each L 5 independently, divalent saturated or unsaturated linear or branched C 1 -C 20 a hydrocarbon chain in which one to three methylene units are optionally and independently replaced by -O- or -NR-; Each R 5 is independently an optionally substituted group selected from a 5- to 10-membered aryl ring and a 3- to 8-membered carbocyclic ring; X 1 is a covalent bond, -O-, or -NR-; X 2 is a covalent bond or an optionally substituted divalent saturated or unsaturated linear or branched C 1 -C 12 a hydrocarbon chain in which one to three methylene units are optionally and independently replaced by -O- or -NR-; X 3 is hydrogen or -Cy B and Cy Bis an optionally substituted ring selected from 3- to 7-membered saturated or partially unsaturated carbocyclyl; phenyl; 3- to 7-membered saturated or partially unsaturated heterocyclyl having 1-3 heteroatoms independently selected from nitrogen, oxygen, and sulfur; and 5- to 6-membered heteroaryl having 1-3 heteroatoms independently selected from nitrogen, oxygen, and sulfur; Each R is independently hydrogen or optionally substituted C 1 -C 6 is an aliphatic group, However, X 3 is hydrogen, R 1 or R 1’ At least one of the [ka] or a pharma- ceutically acceptable salt thereof.

[0112] In some embodiments, the present disclosure provides a compound of formula I, or a pharma- ceutically acceptable salt thereof: Each L 1 and L 1’ is independently -C(O)- or -C(O)O-; Each L 2 and L 2’ are independently a covalent bond, an optionally substituted divalent saturated or unsaturated linear or branched C 1 -C 12 a hydrocarbon chain, or [ka] and Each Cy A is independently an optionally substituted ring selected from phenylene, and a 3- to 7-membered saturated or partially unsaturated carbocyclene; each m is independently 0, 1, or 2; Each L 3 and L 3’are independently a covalent bond, —C(O)O—, —OC(O)—, —O—, or —OC(O)O—; Each R 1 and R 1’ is independently an optionally substituted group, in which 1 to 3 methylene units are optionally and independently replaced by -O- or -NR-, and 1 -C 20 Hydrocarbon chains; 3- to 7-membered saturated or partially unsaturated carbocyclic rings; 1-adamantyl; 2-adamantyl; sterolyl; phenyl; and [ka] is an optionally substituted group selected from Each L 4 independently, divalent saturated or unsaturated linear or branched C 1 -C 6 is a hydrocarbon chain, Each A 1 and A 2 is independently an optionally substituted C 1 -C 20 Aliphatic or -L 5 -R 5 or Or A 1 and A 2 may be taken together with their intervening atoms to form an optionally substituted ring, [ka] During the ceremony, x is selected from 1 or 2; # is L 4 represents the attachment point to Each L 5 independently, divalent saturated or unsaturated linear or branched C 1 -C 20 a hydrocarbon chain in which one to three methylene units are optionally and independently replaced by -O- or -NR-; Each R 5is independently an optionally substituted group selected from a 5- to 10-membered aryl ring and a 3- to 8-membered carbocyclic ring; X 1 is a covalent bond, -O-, or -NR-; X 2 is a covalent bond or an optionally substituted divalent saturated or unsaturated linear or branched C 1 -C 12 a hydrocarbon chain in which one to three methylene units are optionally and independently replaced by -O- or -NR-; X 3 is hydrogen or -Cy B and Cy B is an optionally substituted ring selected from 3- to 7-membered saturated or partially unsaturated carbocyclyl; phenyl; 3- to 7-membered saturated or partially unsaturated heterocyclyl having 1-3 heteroatoms independently selected from nitrogen, oxygen, and sulfur; and 5- to 6-membered heteroaryl having 1-3 heteroatoms independently selected from nitrogen, oxygen, and sulfur; Each R is independently hydrogen or optionally substituted C 1 -C 6 is an aliphatic group, However, X 3 is hydrogen, R 1 or R 1’ At least one of the [ka] or a pharma- ceutically acceptable salt thereof.

[0113] In some embodiments, the present disclosure provides a compound of formula (II): [ka] or a pharma- ceutically acceptable salt thereof, wherein L 1 , L 1’ , L 2 , L 2’ , L3 , L 3’ , R 1 , R 1’ , X 1 , X 2 , and Cy B is as defined above for formula I, and is as described in the classes and subclasses herein, both alone and in combination, or a pharma- ceutically acceptable salt thereof.

[0114] In some embodiments, the present disclosure provides a compound of formula (III): [ka] or a pharma- ceutically acceptable salt thereof, wherein L 1 , L 1’ , L 2 , L 2’ , L 3 , L 3’ , L 4 , A 1 , A 2 , R 1’ , X 1 , and X 2 is as defined above for formula I, and is as described in the classes and subclasses herein, both alone and in combination, or a pharma- ceutically acceptable salt thereof.

[0115] In some embodiments, the present disclosure provides a compound of formula (IV-A): [ka] or a pharma- ceutically acceptable salt thereof, wherein L 2 , L 2’ , L 3 , L 3’ , L 4 , A 1 , A 2 , R 1’ , X 1 , X 2 , and Cy Bis as defined above for formula I, and is as described in the classes and subclasses herein, both alone and in combination, or a pharma- ceutically acceptable salt thereof.

[0116] In some embodiments, the present disclosure provides a compound of formula (IV-B): [ka] or a pharma- ceutically acceptable salt thereof, wherein L 2 , L 2’ , L 3 , L 3’ , L 4 , A 1 , A 2 , R 1’ , X 1 , and X 2 is as defined above for formula I, and is as described in the classes and subclasses herein, both alone and in combination, or a pharma- ceutically acceptable salt thereof.

[0117] In some embodiments, the present disclosure provides a compound of formula (VA): [ka] or a pharma- ceutically acceptable salt thereof, wherein L 2 , L 2’ , L 3 , L 3’ , L 4 , A 1 , A 2 , X 1 , X 2 , and Cy B is as defined above for formula I, and is as described in the classes and subclasses herein, both alone and in combination, or a pharma- ceutically acceptable salt thereof.

[0118] In some embodiments, the present disclosure provides a compound of formula (VB): [ka] or a pharma- ceutically acceptable salt thereof, wherein L 2 , L 2’ , L 3 , L 3’ , L 4 , A 1 , A 2 , X 1 , and X 2 is as defined above for formula I, and is as described in the classes and subclasses herein, both alone and in combination, or a pharma- ceutically acceptable salt thereof.

[0119] In some embodiments, the present disclosure provides a compound of formula (VI-A) or (VI-B): [ka] or a pharma- ceutically acceptable salt thereof, wherein L 2 , L 2’ , L 3 , L 3’ , R 1 , R 1’ , X 2 , and Cy B is as defined above for formula I, and both alone and in combination, are as described in the classes and subclasses herein, or a pharma- ceutically acceptable salt thereof. In some embodiments, the disclosure provides a compound of formula (VI-A) or a pharma- ceutically acceptable salt thereof. In some embodiments, the disclosure provides a compound of formula (VI-B) or a pharma- ceutically acceptable salt thereof.

[0120] In some embodiments, the present disclosure provides a compound of formula (VII-A) or (VII-B): [ka] or a pharma- ceutically acceptable salt thereof, wherein L 2 , L 2’ , L 3 , L 3’ , L4 , R 1’ , A 1 , A 2 , and X 2 is as defined above for formula I, and both alone and in combination, are as described in the classes and subclasses herein, or a pharma- ceutically acceptable salt thereof. In some embodiments, the present disclosure provides a compound of formula (VII-A) or a pharma- ceutically acceptable salt thereof. In some embodiments, the present disclosure provides a compound of formula (VII-B) or a pharma- ceutically acceptable salt thereof.

[0121] In some embodiments, the present disclosure provides a compound of formula (VIII-A), (VIII-B), (VIII-C), or (VIII-D): [ka] or a pharma- ceutically acceptable salt thereof, wherein L 2’ , L 3’ , R 1’ , L 4 , A 1 , A 2 , X 2 , and Cy B is defined above for Formula I, and both alone and in combination, is as described in the classes and subclasses herein, or a pharma- ceutically acceptable salt thereof. In some embodiments, the present disclosure provides a compound of formula (VIII-A), or a pharma- ceutically acceptable salt thereof. In some embodiments, the present disclosure provides a compound of formula (VIII-B), or a pharma- ceutically acceptable salt thereof. In some embodiments, the present disclosure provides a compound of formula (VIII-C), or a pharma- ceutically acceptable salt thereof. In some embodiments, the present disclosure provides a compound of formula (VIII-D), or a pharma- ceutically acceptable salt thereof.

[0122] In some embodiments, the disclosure provides a compound of formula (IX-A), (IX-B), (IX-C), or (IX-D): [ka] or a pharma- ceutically acceptable salt thereof, wherein L 2’ , L 3’ , R 1’ , L 4 , A 1 , A 2 , and X 2 is defined above for Formula I, and both alone and in combination, is as described in the classes and subclasses herein, or a pharma- ceutically acceptable salt thereof. In some embodiments, the present disclosure provides a compound of formula (IX-A) or a pharma- ceutically acceptable salt thereof. In some embodiments, the present disclosure provides a compound of formula (IX-B) or a pharma- ceutically acceptable salt thereof. In some embodiments, the present disclosure provides a compound of formula (IX-C) or a pharma- ceutically acceptable salt thereof. In some embodiments, the present disclosure provides a compound of formula (IX-D) or a pharma- ceutically acceptable salt thereof.

[0123] In some embodiments of any of the formulas described herein, each L 1 and L 1’ is independently -C(O)- or -C(O)O-. 1 In some embodiments, L 1 is -C(O)O-. In some embodiments, L 1’ In some embodiments, L 1’ is -C(O)O-. In some embodiments, L 1 and L 1’ Both of are -C(O)-.

[0124] In some embodiments of any of the formulas described herein, each L 2 and L 2’ are independently a covalent bond, an optionally substituted divalent saturated or unsaturated linear or branched C 1 -C 12 a hydrocarbon chain, or [ka] Throughout this disclosure, [ka] is -(CH 2 ) m -Cy A -(CH 2 ) m - are intended to refer to the same moiety as , and it will be understood that they may be used interchangeably.

[0125] In some embodiments, L 2 is a covalent bond. In some embodiments, L 2 is an optionally substituted divalent saturated or unsaturated linear or branched C 1 -C 12 In some embodiments, L 2 is an optionally substituted divalent saturated or unsaturated linear or branched C 1 -C 9 In some embodiments, L 2 is an optionally substituted divalent saturated or unsaturated linear or branched C 1 -C 6 In some embodiments, L 2 is an optionally substituted divalent saturated or unsaturated linear or branched C 1 -C 3 In some embodiments, L 2 is optionally substituted, -CH 2 -, -CH 2 CH 2 -, -CH 2 CH 2 CH 2 -, -CH 2 CH 2 CH 2 CH 2 -, -CH 2 CH 2 CH 2 CH 2 CH 2 - or -CH 2 CH 2 CH 2 CH2 CH 2 CH 2 In some embodiments, L 2 is an optionally substituted -CH 2 In some embodiments, L 2 is an optionally substituted -CH 2 CH 2 In some embodiments, L 2 is an optionally substituted -CH 2 CH 2 CH 2 In some embodiments, L 2 is an optionally substituted -CH 2 CH 2 CH 2 CH 2 In some embodiments, L 2 is an optionally substituted -CH 2 CH 2 CH 2 CH 2 CH 2 In some embodiments, L 2 is an optionally substituted -CH 2 CH 2 CH 2 CH 2 CH 2 CH 2 In some embodiments, L 2 teeth, [ka] It is.

[0126] In some embodiments, L 2’ is a covalent bond. In some embodiments, L 2’ is an optionally substituted divalent saturated or unsaturated linear or branched C 1 -C 12 In some embodiments, L 2’ is an optionally substituted divalent saturated or unsaturated linear or branched C 1 -C 9In some embodiments, L 2’ is an optionally substituted divalent saturated or unsaturated linear or branched C 1 -C 6 In some embodiments, L 2’ is an optionally substituted divalent saturated or unsaturated linear or branched C 1 -C 3 In some embodiments, L 2’ is optionally substituted, -CH 2 -, -CH 2 CH 2 -, -CH 2 CH 2 CH 2 -, -CH 2 CH 2 CH 2 CH 2 -, -CH 2 CH 2 CH 2 CH 2 CH 2 - or -CH 2 CH 2 CH 2 CH 2 CH 2 CH 2 In some embodiments, L 2’ is an optionally substituted -CH 2 In some embodiments, L 2’ is an optionally substituted -CH 2 CH 2 In some embodiments, L 2’ is an optionally substituted -CH 2 CH 2 CH 2 In some embodiments, L 2’ is an optionally substituted -CH 2 CH 2 CH 2 CH 2 In some embodiments, L 2’ is an optionally substituted -CH 2 CH 2 CH 2 CH 2 CH2 In some embodiments, L 2’ is an optionally substituted -CH 2 CH 2 CH 2 CH 2 CH 2 CH 2 In some embodiments, L 2’ teeth, [ka] It is.

[0127] In some embodiments of any formula described herein, each m is independently 0, 1, or 2. In some embodiments, at least one m is 0. In some embodiments, at least one m is 1. In some embodiments, at least one m is 2. In some embodiments, both occurrences of m are 0. In some embodiments, both occurrences of m are 1. In some embodiments, both occurrences of m are 2.

[0128] In some embodiments of any of the formulas described herein, Cy A is an optionally substituted ring selected from phenylene and 3- to 7-membered saturated or partially unsaturated carbocyclene. A is optionally substituted phenylene. In some embodiments, Cy A teeth, [ka] In some embodiments, Cy A teeth, [ka] In some embodiments, Cy A teeth, [ka] In some embodiments, Cy A is an optionally substituted 3- to 7-membered saturated or partially unsaturated carbocyclene. A is an optionally substituted 5-6 membered saturated or partially unsaturated carbocyclene. A is an optionally substituted 5-6 membered saturated carbocyclene. In some embodiments, Cy A is optionally substituted cyclohexylene. In some embodiments, Cy A teeth, [ka] In some embodiments, Cy A teeth, [ka] In some embodiments, Cy A teeth, [ka] It is.

[0129] In some embodiments of any of the formulas described herein, each L 3 and L 3’ is independently a covalent bond, -C(O)O-, -OC(O)-, -O-, or -OC(O)O-. 3 is a covalent bond. In some embodiments, L 3 is -C(O)O-. In some embodiments, L 3 In some embodiments, L 3 In some embodiments, L 3 is -OC(O)O-. In some embodiments, L 3’ is a covalent bond. In some embodiments, L 3’ is -C(O)O-. In some embodiments, L 3’In some embodiments, L 3’ In some embodiments, L 3’ is -OC(O)O-.

[0130] In some embodiments of any of the formulas described herein, each R 1 and R 1’ is independently an optionally substituted group, wherein one to three methylene units are optionally and independently replaced by -O- or -NR-, and is a saturated or unsaturated, linear or branched C 1 -C 20 Hydrocarbon chains; 3- to 12-membered saturated or partially unsaturated carbocyclic rings; 1-adamantyl; 2-adamantyl; sterolyl; phenyl; and [ka] In some embodiments, each R 1 and R 1’ is independently an optionally substituted group, wherein one to three methylene units are optionally and independently replaced by -O- or -NR-, and is a saturated or unsaturated, linear or branched C 1 -C 20 Hydrocarbon chains; 3- to 7-membered saturated or partially unsaturated carbocyclic rings; 1-adamantyl; 2-adamantyl; sterolyl; phenyl; and [ka] is an optionally substituted group selected from

[0131] In some embodiments, R 1 is an optionally substituted saturated or unsaturated linear or branched C 1 -C 20 In some embodiments, R is a hydrocarbon chain in which one to three methylene units are optionally and independently replaced with -O- or -NR-. 1is an optionally substituted saturated or unsaturated linear or branched C 1 -C 15 In some embodiments, R is a hydrocarbon chain in which one to three methylene units are optionally and independently replaced with -O- or -NR-. 1 is an optionally substituted saturated or unsaturated linear or branched C 1 -C 12 In some embodiments, R is a hydrocarbon chain in which one to three methylene units are optionally and independently replaced with -O- or -NR-. 1 is an optionally substituted saturated or unsaturated linear or branched C 1 -C 9 In some embodiments, R is a hydrocarbon chain in which one to three methylene units are optionally and independently replaced with -O- or -NR-. 1 is an optionally substituted saturated or unsaturated linear or branched C 1 -C 6 In some embodiments, R is a hydrocarbon chain in which one to two methylene units are optionally and independently replaced with -O- or -NR-. 1 is an optionally substituted saturated or unsaturated linear or branched C 6 -C 20 In some embodiments, R is a hydrocarbon chain in which one to three methylene units are optionally and independently replaced with -O- or -NR-. 1 is an optionally substituted saturated or unsaturated linear or branched C 9 -C 20 In some embodiments, R is a hydrocarbon chain in which one to three methylene units are optionally and independently replaced with -O- or -NR-. 1 is an optionally substituted saturated or unsaturated linear or branched C 12 -C 20 In some embodiments, R is a hydrocarbon chain in which one to three methylene units are optionally and independently replaced with -O- or -NR-. 1 is an optionally substituted saturated or unsaturated linear or branched C 15 -C20 In some embodiments, R is a hydrocarbon chain in which one to three methylene units are optionally and independently replaced with -O- or -NR-. 1 is an optionally substituted saturated or unsaturated linear or branched C 6 -C 15 In some embodiments, R is a hydrocarbon chain in which one to three methylene units are optionally and independently replaced with -O- or -NR-. 1 is an optionally substituted saturated or unsaturated linear or branched C 8 -C 17 A hydrocarbon chain in which one to three methylene units are optionally and independently replaced by -O- or -NR-.

[0132] In some embodiments, R 1 is an optionally substituted saturated or unsaturated linear or branched C 6 In some embodiments, R is a hydrocarbon chain in which one to two methylene units are optionally and independently replaced with -O- or -NR-. 1 is an optionally substituted saturated or unsaturated linear or branched C 7 In some embodiments, R is a hydrocarbon chain in which one to two methylene units are optionally and independently replaced with -O- or -NR-. 1 is an optionally substituted saturated or unsaturated linear or branched C 8 In some embodiments, R is a hydrocarbon chain in which one to two methylene units are optionally and independently replaced with -O- or -NR-. 1 is an optionally substituted saturated or unsaturated linear or branched C 9 In some embodiments, R is a hydrocarbon chain in which one to three methylene units are optionally and independently replaced with -O- or -NR-. 1 is an optionally substituted saturated or unsaturated linear or branched C 10 In some embodiments, R is a hydrocarbon chain in which one to three methylene units are optionally and independently replaced with -O- or -NR-.1 is an optionally substituted saturated or unsaturated linear or branched C 11 In some embodiments, R is a hydrocarbon chain in which one to three methylene units are optionally and independently replaced with -O- or -NR-. 1 is an optionally substituted saturated or unsaturated linear or branched C 12 In some embodiments, R is a hydrocarbon chain in which one to three methylene units are optionally and independently replaced with -O- or -NR-. 1 is an optionally substituted saturated or unsaturated linear or branched C 13 In some embodiments, R is a hydrocarbon chain in which one to three methylene units are optionally and independently replaced with -O- or -NR-. 1 is an optionally substituted saturated or unsaturated linear or branched C 14 In some embodiments, R is a hydrocarbon chain in which one to three methylene units are optionally and independently replaced with -O- or -NR-. 1 is an optionally substituted saturated or unsaturated linear or branched C 15 In some embodiments, R is a hydrocarbon chain in which one to three methylene units are optionally and independently replaced with -O- or -NR-. 1 is an optionally substituted saturated or unsaturated linear or branched C 16 In some embodiments, R is a hydrocarbon chain in which one to three methylene units are optionally and independently replaced with -O- or -NR-. 1 is an optionally substituted saturated or unsaturated linear or branched C 17 In some embodiments, R is a hydrocarbon chain in which one to three methylene units are optionally and independently replaced with -O- or -NR-. 1 is an optionally substituted saturated or unsaturated linear or branched C 18 In some embodiments, R is a hydrocarbon chain in which one to three methylene units are optionally and independently replaced with -O- or -NR-. 1is an optionally substituted saturated or unsaturated linear or branched C 19 In some embodiments, R is a hydrocarbon chain in which one to three methylene units are optionally and independently replaced with -O- or -NR-. 1 is an optionally substituted saturated or unsaturated linear or branched C 20 A hydrocarbon chain in which one to three methylene units are optionally and independently replaced by -O- or -NR-.

[0133] In some embodiments, R 1 is an optionally substituted saturated or unsaturated linear or branched C 1 -C 20 In some embodiments, R is a hydrocarbon chain, in which one methylene unit is optionally and independently replaced with -O-. 1 is an optionally substituted saturated or unsaturated linear or branched C 1 -C 15 In some embodiments, R is a hydrocarbon chain, in which one methylene unit is optionally and independently replaced with -O-. 1 is an optionally substituted saturated or unsaturated linear or branched C 1 -C 12 In some embodiments, R is a hydrocarbon chain, in which one methylene unit is optionally and independently replaced with -O-. 1 is an optionally substituted saturated or unsaturated linear or branched C 1 -C 9 In some embodiments, R is a hydrocarbon chain, in which one methylene unit is optionally and independently replaced with -O-. 1 is an optionally substituted saturated or unsaturated linear or branched C 1 -C 6 In some embodiments, R is a hydrocarbon chain, in which one methylene unit is optionally and independently replaced with -O-. 1 is an optionally substituted saturated or unsaturated linear or branched C 6 -C 20In some embodiments, R is a hydrocarbon chain, in which one methylene unit is optionally and independently replaced with -O-. 1 is an optionally substituted saturated or unsaturated linear or branched C 9 -C 20 In some embodiments, R is a hydrocarbon chain, in which one methylene unit is optionally and independently replaced with -O-. 1 is an optionally substituted saturated or unsaturated linear or branched C 12 -C 20 In some embodiments, R is a hydrocarbon chain, in which one methylene unit is optionally and independently replaced with -O-. 1 is an optionally substituted saturated or unsaturated linear or branched C 15 -C 20 In some embodiments, R is a hydrocarbon chain, in which one methylene unit is optionally and independently replaced with -O-. 1 is an optionally substituted saturated or unsaturated linear or branched C 6 -C 15 In some embodiments, R is a hydrocarbon chain, in which one methylene unit is optionally and independently replaced with -O-. 1 is an optionally substituted saturated or unsaturated linear or branched C 8 -C 17 A hydrocarbon chain in which one methylene unit is optionally and independently replaced with -O-.

[0134] In some embodiments, R 1 is an optionally substituted saturated or unsaturated linear or branched C 6 In some embodiments, R is a hydrocarbon chain, in which one methylene unit is optionally and independently replaced with -O-. 1 is an optionally substituted saturated or unsaturated linear or branched C 7 In some embodiments, R is a hydrocarbon chain, in which one methylene unit is optionally and independently replaced with -O-. 1is an optionally substituted saturated or unsaturated linear or branched C 8 In some embodiments, R is a hydrocarbon chain, in which one methylene unit is optionally and independently replaced with -O-. 1 is an optionally substituted saturated or unsaturated linear or branched C 9 In some embodiments, R is a hydrocarbon chain, in which one methylene unit is optionally and independently replaced with -O-. 1 is an optionally substituted saturated or unsaturated linear or branched C 10 In some embodiments, R is a hydrocarbon chain, in which one methylene unit is optionally and independently replaced with -O-. 1 is an optionally substituted saturated or unsaturated linear or branched C 11 In some embodiments, R is a hydrocarbon chain, in which one methylene unit is optionally and independently replaced with -O-. 1 is an optionally substituted saturated or unsaturated linear or branched C 12 In some embodiments, R is a hydrocarbon chain in which one methylene unit is optionally and independently replaced with -O-. 1 is an optionally substituted saturated or unsaturated linear or branched C 13 In some embodiments, R is a hydrocarbon chain, in which one methylene unit is optionally and independently replaced with -O-. 1 is an optionally substituted saturated or unsaturated linear or branched C 14 In some embodiments, R is a hydrocarbon chain, in which one methylene unit is optionally and independently replaced with -O-. 1 is an optionally substituted saturated or unsaturated linear or branched C 15 In some embodiments, R is a hydrocarbon chain, in which one methylene unit is optionally and independently replaced with -O-. 1 is an optionally substituted saturated or unsaturated linear or branched C 16 In some embodiments, R is a hydrocarbon chain, in which one methylene unit is optionally and independently replaced with -O-.1 is an optionally substituted saturated or unsaturated linear or branched C 17 In some embodiments, R is a hydrocarbon chain, in which one methylene unit is optionally and independently replaced with -O-. 1 is an optionally substituted saturated or unsaturated linear or branched C 18 In some embodiments, R is a hydrocarbon chain, in which one methylene unit is optionally and independently replaced with -O-. 1 is an optionally substituted saturated or unsaturated linear or branched C 19 In some embodiments, R is a hydrocarbon chain, in which one methylene unit is optionally and independently replaced with -O-. 1 is an optionally substituted saturated or unsaturated linear or branched C 20 A hydrocarbon chain in which one methylene unit is optionally and independently replaced with -O-.

[0135] In some embodiments, R 1 is an optionally substituted saturated or unsaturated linear or branched C 1 -C 20 In some embodiments, R 1 is an optionally substituted saturated or unsaturated linear or branched C 1 -C 15 In some embodiments, R 1 is an optionally substituted saturated or unsaturated linear or branched C 1 -C 12 In some embodiments, R 1 is an optionally substituted saturated or unsaturated linear or branched C 1 -C 9 In some embodiments, R 1 is an optionally substituted saturated or unsaturated linear or branched C 1 -C 6 In some embodiments, R 1 is an optionally substituted saturated or unsaturated linear or branched C 6-C 20 In some embodiments, R 1 is an optionally substituted saturated or unsaturated linear or branched C 9 -C 20 In some embodiments, R 1 is an optionally substituted saturated or unsaturated linear or branched C 12 -C 20 In some embodiments, R 1 is an optionally substituted saturated or unsaturated linear or branched C 15 -C 20 In some embodiments, R 1 is an optionally substituted saturated or unsaturated linear or branched C 6 -C 15 In some embodiments, R 1 is an optionally substituted saturated or unsaturated linear or branched C 8 -C 17 It is a hydrocarbon chain.

[0136] In some embodiments, R 1 is an optionally substituted saturated or unsaturated linear or branched C 6 In some embodiments, R 1 is an optionally substituted saturated or unsaturated linear or branched C 7 In some embodiments, R 1 is an optionally substituted saturated or unsaturated linear or branched C 8 In some embodiments, R 1 is an optionally substituted saturated or unsaturated linear or branched C 9 In some embodiments, R 1 is an optionally substituted saturated or unsaturated linear or branched C 10 In some embodiments, R 1 is an optionally substituted saturated or unsaturated linear or branched C 11 In some embodiments, R 1is an optionally substituted saturated or unsaturated linear or branched C 12 In some embodiments, R 1 is an optionally substituted saturated or unsaturated linear or branched C 13 In some embodiments, R 1 is an optionally substituted saturated or unsaturated linear or branched C 14 In some embodiments, R 1 is an optionally substituted saturated or unsaturated linear or branched C 15 In some embodiments, R 1 is an optionally substituted saturated or unsaturated linear or branched C 16 In some embodiments, R 1 is an optionally substituted saturated or unsaturated linear or branched C 17 In some embodiments, R 1 is an optionally substituted saturated or unsaturated linear or branched C 18 In some embodiments, R 1 is an optionally substituted saturated or unsaturated linear or branched C 19 In some embodiments, R 1 is an optionally substituted saturated or unsaturated linear or branched C 20 It is a hydrocarbon chain.

[0137] In some embodiments, R 1 is a saturated or unsaturated linear or branched C optionally substituted with 1 to 6 halogen atoms; 1 -C 20 In some embodiments, R 1 is a saturated or unsaturated linear or branched C optionally substituted with 1 to 6 halogen atoms; 1 -C 15 In some embodiments, R 1 is a saturated or unsaturated linear or branched C optionally substituted with 1 to 6 halogen atoms; 1 -C 12In some embodiments, R 1 is a saturated or unsaturated linear or branched C optionally substituted with 1 to 6 halogen atoms; 1 -C 9 In some embodiments, R 1 is a saturated or unsaturated linear or branched C optionally substituted with 1 to 6 halogen atoms; 1 -C 6 In some embodiments, R 1 is a saturated or unsaturated linear or branched C optionally substituted with 1 to 6 halogen atoms; 6 -C 20 In some embodiments, R 1 is a saturated or unsaturated linear or branched C optionally substituted with 1 to 6 halogen atoms; 9 -C 20 In some embodiments, R 1 is a saturated or unsaturated linear or branched C optionally substituted with 1 to 6 halogen atoms; 12 -C 20 In some embodiments, R 1 is a saturated or unsaturated linear or branched C optionally substituted with 1 to 6 halogen atoms; 15 -C 20 In some embodiments, R 1 is a saturated or unsaturated linear or branched C optionally substituted with 1 to 6 halogen atoms; 6 -C 15 In some embodiments, R 1 is a saturated or unsaturated linear or branched C optionally substituted with 1 to 6 halogen atoms; 8 -C 17 It is a hydrocarbon chain.

[0138] In some embodiments, R 1 is a saturated or unsaturated linear or branched C optionally substituted with 1 to 6 halogen atoms; 6 In some embodiments, R 1is a saturated or unsaturated linear or branched C optionally substituted with 1 to 6 halogen atoms; 7 In some embodiments, R 1 is a saturated or unsaturated linear or branched C optionally substituted with 1 to 6 halogen atoms; 8 In some embodiments, R 1 is a saturated or unsaturated linear or branched C optionally substituted with 1 to 6 halogen atoms; 9 In some embodiments, R 1 is a saturated or unsaturated linear or branched C optionally substituted with 1 to 6 halogen atoms; 10 In some embodiments, R 1 is a saturated or unsaturated linear or branched C optionally substituted with 1 to 6 halogen atoms; 11 In some embodiments, R 1 is a saturated or unsaturated linear or branched C optionally substituted with 1 to 6 halogen atoms; 12 In some embodiments, R 1 is a saturated or unsaturated linear or branched C optionally substituted with 1 to 6 halogen atoms; 13 In some embodiments, R 1 is a saturated or unsaturated linear or branched C optionally substituted with 1 to 6 halogen atoms; 14 In some embodiments, R 1 is a saturated or unsaturated linear or branched C optionally substituted with 1 to 6 halogen atoms; 15 In some embodiments, R 1 is a saturated or unsaturated linear or branched C optionally substituted with 1 to 6 halogen atoms; 16 In some embodiments, R 1 is a saturated or unsaturated linear or branched C optionally substituted with 1 to 6 halogen atoms; 17 In some embodiments, R 1is a saturated or unsaturated linear or branched C optionally substituted with 1 to 6 halogen atoms; 18 In some embodiments, R 1 is a saturated or unsaturated linear or branched C optionally substituted with 1 to 6 halogen atoms; 19 In some embodiments, R 1 is a saturated or unsaturated linear or branched C optionally substituted with 1 to 6 halogen atoms; 20 It is a hydrocarbon chain.

[0139] In some embodiments, R 1 is a saturated or unsaturated linear or branched C optionally substituted with 1 to 6 fluorine atoms; 1 -C 20 In some embodiments, R 1 is a saturated or unsaturated linear or branched C optionally substituted with 1 to 6 fluorine atoms; 1 -C 15 In some embodiments, R 1 is a saturated or unsaturated linear or branched C optionally substituted with 1 to 6 fluorine atoms; 1 -C 12 In some embodiments, R 1 is a saturated or unsaturated linear or branched C optionally substituted with 1 to 6 fluorine atoms; 1 -C 9 In some embodiments, R 1 is a saturated or unsaturated linear or branched C optionally substituted with 1 to 6 fluorine atoms; 1 -C 6 In some embodiments, R 1 is a saturated or unsaturated linear or branched C optionally substituted with 1 to 6 fluorine atoms; 6 -C 20 In some embodiments, R 1 is a saturated or unsaturated linear or branched C optionally substituted with 1 to 6 fluorine atoms; 9 -C 20In some embodiments, R 1 is a saturated or unsaturated linear or branched C optionally substituted with 1 to 6 fluorine atoms; 12 -C 20 In some embodiments, R 1 is a saturated or unsaturated linear or branched C optionally substituted with 1 to 6 fluorine atoms; 15 -C 20 In some embodiments, R 1 is a saturated or unsaturated linear or branched C optionally substituted with 1 to 6 fluorine atoms; 6 -C 15 In some embodiments, R 1 is a saturated or unsaturated linear or branched C optionally substituted with 1 to 6 fluorine atoms; 8 -C 17 It is a hydrocarbon chain.

[0140] In some embodiments, R 1 is a saturated or unsaturated linear or branched C optionally substituted with 1 to 6 fluorine atoms; 6 In some embodiments, R 1 is a saturated or unsaturated linear or branched C optionally substituted with 1 to 6 fluorine atoms; 7 In some embodiments, R 1 is a saturated or unsaturated linear or branched C optionally substituted with 1 to 6 fluorine atoms; 8 In some embodiments, R 1 is a saturated or unsaturated linear or branched C optionally substituted with 1 to 6 fluorine atoms; 9 In some embodiments, R 1 is a saturated or unsaturated linear or branched C optionally substituted with 1 to 6 fluorine atoms; 10 In some embodiments, R 1 is a saturated or unsaturated linear or branched C optionally substituted with 1 to 6 fluorine atoms; 11 In some embodiments, R 1is a saturated or unsaturated linear or branched C optionally substituted with 1 to 6 fluorine atoms; 12 In some embodiments, R 1 is a saturated or unsaturated linear or branched C optionally substituted with 1 to 6 fluorine atoms; 13 In some embodiments, R 1 is a saturated or unsaturated linear or branched C optionally substituted with 1 to 6 fluorine atoms; 14 In some embodiments, R 1 is a saturated or unsaturated linear or branched C optionally substituted with 1 to 6 fluorine atoms; 15 In some embodiments, R 1 is a saturated or unsaturated linear or branched C optionally substituted with 1 to 6 fluorine atoms; 16 In some embodiments, R 1 is a saturated or unsaturated linear or branched C optionally substituted with 1 to 6 fluorine atoms; 17 In some embodiments, R 1 is a saturated or unsaturated linear or branched C optionally substituted with 1 to 6 fluorine atoms; 18 In some embodiments, R 1 is a saturated or unsaturated linear or branched C optionally substituted with 1 to 6 fluorine atoms; 19 In some embodiments, R 1 is a saturated or unsaturated linear or branched C optionally substituted with 1 to 6 fluorine atoms; 20 It is a hydrocarbon chain.

[0141] In some embodiments, R 1 is a saturated or unsaturated linear or branched C substituted with 1 to 6 fluorine atoms. 1 -C 20 In some embodiments, R 1 is a saturated or unsaturated linear or branched C substituted with 1 to 6 fluorine atoms. 1 -C 15 In some embodiments, R1 is a saturated or unsaturated linear or branched C substituted with 1 to 6 fluorine atoms. 1 -C 12 In some embodiments, R 1 is a saturated or unsaturated linear or branched C substituted with 1 to 6 fluorine atoms. 1 -C 9 In some embodiments, R 1 is a saturated or unsaturated linear or branched C substituted with 1 to 6 fluorine atoms. 1 -C 6 In some embodiments, R 1 is a saturated or unsaturated linear or branched C substituted with 1 to 6 fluorine atoms. 6 -C 20 In some embodiments, R 1 is a saturated or unsaturated linear or branched C substituted with 1 to 6 fluorine atoms. 9 -C 20 In some embodiments, R 1 is a saturated or unsaturated linear or branched C substituted with 1 to 6 fluorine atoms. 12 -C 20 In some embodiments, R 1 is a saturated or unsaturated linear or branched C substituted with 1 to 6 fluorine atoms. 15 -C 20 In some embodiments, R 1 is a saturated or unsaturated linear or branched C substituted with 1 to 6 fluorine atoms. 6 -C 15 In some embodiments, R 1 is a saturated or unsaturated linear or branched C substituted with 1 to 6 fluorine atoms. 8 -C 17 It is a hydrocarbon chain.

[0142] In some embodiments, R 1 is a saturated or unsaturated linear or branched C substituted with 1 to 6 fluorine atoms. 6 In some embodiments, R 1is a saturated or unsaturated linear or branched C substituted with 1 to 6 fluorine atoms. 7 In some embodiments, R 1 is a saturated or unsaturated linear or branched C substituted with 1 to 6 fluorine atoms. 8 In some embodiments, R 1 is a saturated or unsaturated linear or branched C substituted with 1 to 6 fluorine atoms. 9 In some embodiments, R 1 is a saturated or unsaturated linear or branched C substituted with 1 to 6 fluorine atoms. 10 In some embodiments, R 1 is a saturated or unsaturated linear or branched C substituted with 1 to 6 fluorine atoms. 11 In some embodiments, R 1 is a saturated or unsaturated linear or branched C substituted with 1 to 6 fluorine atoms. 12 In some embodiments, R 1 is a saturated or unsaturated linear or branched C substituted with 1 to 6 fluorine atoms. 13 In some embodiments, R 1 is a saturated or unsaturated linear or branched C substituted with 1 to 6 fluorine atoms. 14 In some embodiments, R 1 is a saturated or unsaturated linear or branched C substituted with 1 to 6 fluorine atoms. 15 In some embodiments, R 1 is a saturated or unsaturated linear or branched C substituted with 1 to 6 fluorine atoms. 16 In some embodiments, R 1 is a saturated or unsaturated linear or branched C substituted with 1 to 6 fluorine atoms. 17 In some embodiments, R 1 is a saturated or unsaturated linear or branched C substituted with 1 to 6 fluorine atoms. 18 In some embodiments, R 1is a saturated or unsaturated linear or branched C substituted with 1 to 6 fluorine atoms. 19 In some embodiments, R 1 is a saturated or unsaturated linear or branched C substituted with 1 to 6 fluorine atoms. 20 It is a hydrocarbon chain.

[0143] In some embodiments, R 1 is a saturated or unsaturated linear or branched C 1 -C 20 In some embodiments, R 1 is a saturated or unsaturated linear or branched C 1 -C 15 In some embodiments, R 1 is a saturated or unsaturated linear or branched C 1 -C 12 In some embodiments, R 1 is a saturated or unsaturated linear or branched C 1 -C 9 In some embodiments, R 1 is a saturated or unsaturated linear or branched C 1 -C 6 In some embodiments, R 1 is a saturated or unsaturated linear or branched C 6 -C 20 In some embodiments, R 1 is a saturated or unsaturated linear or branched C 9 -C 20 In some embodiments, R 1 is a saturated or unsaturated linear or branched C 12 -C 20 In some embodiments, R 1 is a saturated or unsaturated linear or branched C 15 -C 20 In some embodiments, R 1 is a saturated or unsaturated linear or branched C 6 -C 15 In some embodiments, R 1is a saturated or unsaturated linear or branched C 8 -C 17 It is a hydrocarbon chain.

[0144] In some embodiments, R 1 is a saturated or unsaturated linear or branched C 6 In some embodiments, R 1 is a saturated or unsaturated linear or branched C 7 In some embodiments, R 1 is a saturated or unsaturated linear or branched C 8 In some embodiments, R 1 is a saturated or unsaturated linear or branched C 9 In some embodiments, R 1 is a saturated or unsaturated linear or branched C 10 In some embodiments, R 1 is a saturated or unsaturated linear or branched C 11 In some embodiments, R 1 is a saturated or unsaturated linear or branched C 12 In some embodiments, R 1 is a saturated or unsaturated linear or branched C 13 In some embodiments, R 1 is a saturated or unsaturated linear or branched C 14 In some embodiments, R 1 is a saturated or unsaturated linear or branched C 15 In some embodiments, R 1 is a saturated or unsaturated linear or branched C 16 In some embodiments, R 1 is a saturated or unsaturated linear or branched C 17 In some embodiments, R 1 is a saturated or unsaturated linear or branched C 18 In some embodiments, R 1 is a saturated or unsaturated linear or branched C 19In some embodiments, R 1 is a saturated or unsaturated linear or branched C 20 It is a hydrocarbon chain.

[0145] In some embodiments, R 1 is an optionally substituted 3-12 membered saturated or partially unsaturated carbocyclyl. In some embodiments, R 1 is an optionally substituted 3- to 7-membered saturated or partially unsaturated carbocyclyl. In some embodiments, R 1 is an optionally substituted 5-6 membered saturated or partially unsaturated carbocyclyl. In some embodiments, R 1 is optionally substituted cyclopropyl. In some embodiments, R 1 is optionally substituted cyclobutyl. In some embodiments, R 1 is optionally substituted cyclopentyl. In some embodiments, R 1 is optionally substituted cyclohexyl. In some embodiments, R 1 is an optionally substituted cycloheptyl. In some embodiments, R 1 Optionally, C 1 -C 6 Aliphatic (e.g., C 1-6 and cyclohexyl substituted with alkyl.

[0146] In some embodiments, R 1 is an optionally substituted 1-adamantyl. In some embodiments, R 1 is an optionally substituted 2-adamantyl. In some embodiments, R 1 is an optionally substituted sterolyl. In some embodiments, R 1 is optionally substituted phenyl. In some embodiments, R 1 is 1-adamantyl. In some embodiments, R 1 is 2-adamantyl. In some embodiments, R 1is sterolyl. In some embodiments, R 1 is phenyl. In some embodiments, R 1 teeth, [ka] It is.

[0147] In some embodiments, R 1 is selected from the group consisting of: [ka] 1-adamantyl, [ka]

[0148] In some embodiments, R 1 is selected from the group consisting of: [ka] 1-adamantyl, [ka]

[0149] In some embodiments, R 1 is selected from the group consisting of: [ka] 1-adamantyl, [ka]

[0150] In some embodiments, R 1’ is an optionally substituted saturated or unsaturated linear or branched C 1 -C 20In some embodiments, R is a hydrocarbon chain in which one to three methylene units are optionally and independently replaced with -O- or -NR-. 1’ is an optionally substituted saturated or unsaturated linear or branched C 1 -C 15 In some embodiments, R is a hydrocarbon chain in which one to three methylene units are optionally and independently replaced with -O- or -NR-. 1’ is an optionally substituted saturated or unsaturated linear or branched C 1 -C 12 In some embodiments, R is a hydrocarbon chain in which one to three methylene units are optionally and independently replaced with -O- or -NR-. 1’ is an optionally substituted saturated or unsaturated linear or branched C 1 -C 9 In some embodiments, R is a hydrocarbon chain in which one to three methylene units are optionally and independently replaced with -O- or -NR-. 1’ is an optionally substituted saturated or unsaturated linear or branched C 1 -C 6 In some embodiments, R is a hydrocarbon chain in which one to two methylene units are optionally and independently replaced with -O- or -NR-. 1’ is an optionally substituted saturated or unsaturated linear or branched C 6 -C 20 In some embodiments, R is a hydrocarbon chain in which one to three methylene units are optionally and independently replaced with -O- or -NR-. 1’ is an optionally substituted saturated or unsaturated linear or branched C 9 -C 20 In some embodiments, R is a hydrocarbon chain in which one to three methylene units are optionally and independently replaced with -O- or -NR-. 1’ is an optionally substituted saturated or unsaturated linear or branched C 12 -C 20In some embodiments, R is a hydrocarbon chain in which one to three methylene units are optionally and independently replaced with -O- or -NR-. 1’ is an optionally substituted saturated or unsaturated linear or branched C 15 -C 20 In some embodiments, R is a hydrocarbon chain in which one to three methylene units are optionally and independently replaced with -O- or -NR-. 1’ is an optionally substituted saturated or unsaturated linear or branched C 6 -C 15 In some embodiments, R is a hydrocarbon chain in which one to three methylene units are optionally and independently replaced with -O- or -NR-. 1’ is an optionally substituted saturated or unsaturated linear or branched C 8 -C 17 A hydrocarbon chain in which one to three methylene units are optionally and independently replaced by -O- or -NR-.

[0151] In some embodiments, R 1’ is an optionally substituted saturated or unsaturated linear or branched C 6 In some embodiments, R is a hydrocarbon chain in which one to two methylene units are optionally and independently replaced with -O- or -NR-. 1’ is an optionally substituted saturated or unsaturated linear or branched C 7 In some embodiments, R is a hydrocarbon chain in which one to two methylene units are optionally and independently replaced with -O- or -NR-. 1’ is an optionally substituted saturated or unsaturated linear or branched C 8 In some embodiments, R is a hydrocarbon chain in which one to two methylene units are optionally and independently replaced with -O- or -NR-. 1’ is an optionally substituted saturated or unsaturated linear or branched C 9 In some embodiments, R is a hydrocarbon chain in which one to three methylene units are optionally and independently replaced with -O- or -NR-.1’ is an optionally substituted saturated or unsaturated linear or branched C 10 In some embodiments, R is a hydrocarbon chain in which one to three methylene units are optionally and independently replaced with -O- or -NR-. 1’ is an optionally substituted saturated or unsaturated linear or branched C 11 In some embodiments, R is a hydrocarbon chain in which one to three methylene units are optionally and independently replaced with -O- or -NR-. 1’ is an optionally substituted saturated or unsaturated linear or branched C 12 In some embodiments, R is a hydrocarbon chain in which one to three methylene units are optionally and independently replaced with -O- or -NR-. 1’ is an optionally substituted saturated or unsaturated linear or branched C 13 In some embodiments, R is a hydrocarbon chain in which one to three methylene units are optionally and independently replaced with -O- or -NR-. 1’ is an optionally substituted saturated or unsaturated linear or branched C 14 In some embodiments, R is a hydrocarbon chain in which one to three methylene units are optionally and independently replaced with -O- or -NR-. 1’ is an optionally substituted saturated or unsaturated linear or branched C 15 In some embodiments, R is a hydrocarbon chain in which one to three methylene units are optionally and independently replaced with -O- or -NR-. 1’ is an optionally substituted saturated or unsaturated linear or branched C 16 In some embodiments, R is a hydrocarbon chain in which one to three methylene units are optionally and independently replaced with -O- or -NR-. 1’ is an optionally substituted saturated or unsaturated linear or branched C 17 In some embodiments, R is a hydrocarbon chain in which one to three methylene units are optionally and independently replaced with -O- or -NR-. 1’is an optionally substituted saturated or unsaturated linear or branched C 18 In some embodiments, R is a hydrocarbon chain in which one to three methylene units are optionally and independently replaced with -O- or -NR-. 1’ is an optionally substituted saturated or unsaturated linear or branched C 19 In some embodiments, R is a hydrocarbon chain in which one to three methylene units are optionally and independently replaced with -O- or -NR-. 1’ is an optionally substituted saturated or unsaturated linear or branched C 20 A hydrocarbon chain in which one to three methylene units are optionally and independently replaced by -O- or -NR-.

[0152] In some embodiments, R 1’ is an optionally substituted saturated or unsaturated linear or branched C 1 -C 20 In some embodiments, R is a hydrocarbon chain, in which one methylene unit is optionally and independently replaced with -O-. 1’ is an optionally substituted saturated or unsaturated linear or branched C 1 -C 15 In some embodiments, R is a hydrocarbon chain, in which one methylene unit is optionally and independently replaced with -O-. 1’ is an optionally substituted saturated or unsaturated linear or branched C 1 -C 12 In some embodiments, R is a hydrocarbon chain, in which one methylene unit is optionally and independently replaced with -O-. 1’ is an optionally substituted saturated or unsaturated linear or branched C 1 -C 9 In some embodiments, R is a hydrocarbon chain, in which one methylene unit is optionally and independently replaced with -O-. 1’ is an optionally substituted saturated or unsaturated linear or branched C 1 -C 6In some embodiments, R is a hydrocarbon chain, in which one methylene unit is optionally and independently replaced with -O-. 1’ is an optionally substituted saturated or unsaturated linear or branched C 6 -C 20 In some embodiments, R is a hydrocarbon chain, in which one methylene unit is optionally and independently replaced with -O-. 1’ is an optionally substituted saturated or unsaturated linear or branched C 9 -C 20 In some embodiments, R is a hydrocarbon chain, in which one methylene unit is optionally and independently replaced with -O-. 1’ is an optionally substituted saturated or unsaturated linear or branched C 12 -C 20 In some embodiments, R is a hydrocarbon chain, in which one methylene unit is optionally and independently replaced with -O-. 1’ is an optionally substituted saturated or unsaturated linear or branched C 15 -C 20 In some embodiments, R is a hydrocarbon chain, in which one methylene unit is optionally and independently replaced with -O-. 1’ is an optionally substituted saturated or unsaturated linear or branched C 6 -C 15 In some embodiments, R is a hydrocarbon chain, in which one methylene unit is optionally and independently replaced with -O-. 1’ is an optionally substituted saturated or unsaturated linear or branched C 8 -C 17 A hydrocarbon chain in which one methylene unit is optionally and independently replaced with -O-.

[0153] In some embodiments, R 1’ is an optionally substituted saturated or unsaturated linear or branched C 6 In some embodiments, R is a hydrocarbon chain, in which one methylene unit is optionally and independently replaced with -O-. 1’is an optionally substituted saturated or unsaturated linear or branched C 7 In some embodiments, R is a hydrocarbon chain, in which one methylene unit is optionally and independently replaced with -O-. 1’ is an optionally substituted saturated or unsaturated linear or branched C 8 In some embodiments, R is a hydrocarbon chain, in which one methylene unit is optionally and independently replaced with -O-. 1’ is an optionally substituted saturated or unsaturated linear or branched C 9 In some embodiments, R is a hydrocarbon chain, in which one methylene unit is optionally and independently replaced with -O-. 1’ is an optionally substituted saturated or unsaturated linear or branched C 10 In some embodiments, R is a hydrocarbon chain, in which one methylene unit is optionally and independently replaced with -O-. 1’ is an optionally substituted saturated or unsaturated linear or branched C 11 In some embodiments, R is a hydrocarbon chain, in which one methylene unit is optionally and independently replaced with -O-. 1’ is an optionally substituted saturated or unsaturated linear or branched C 12 In some embodiments, R is a hydrocarbon chain in which one methylene unit is optionally and independently replaced with -O-. 1’ is an optionally substituted saturated or unsaturated linear or branched C 13 In some embodiments, R is a hydrocarbon chain, in which one methylene unit is optionally and independently replaced with -O-. 1’ is an optionally substituted saturated or unsaturated linear or branched C 14 In some embodiments, R is a hydrocarbon chain, in which one methylene unit is optionally and independently replaced with -O-. 1’ is an optionally substituted saturated or unsaturated linear or branched C 15 In some embodiments, R is a hydrocarbon chain, in which one methylene unit is optionally and independently replaced with -O-.1’ is an optionally substituted saturated or unsaturated linear or branched C 16 In some embodiments, R is a hydrocarbon chain, in which one methylene unit is optionally and independently replaced with -O-. 1’ is an optionally substituted saturated or unsaturated linear or branched C 17 In some embodiments, R is a hydrocarbon chain, in which one methylene unit is optionally and independently replaced with -O-. 1’ is an optionally substituted saturated or unsaturated linear or branched C 18 In some embodiments, R is a hydrocarbon chain, in which one methylene unit is optionally and independently replaced with -O-. 1’ is an optionally substituted saturated or unsaturated linear or branched C 19 In some embodiments, R is a hydrocarbon chain, in which one methylene unit is optionally and independently replaced with -O-. 1’ is an optionally substituted saturated or unsaturated linear or branched C 20 A hydrocarbon chain in which one methylene unit is optionally and independently replaced with -O-.

[0154] In some embodiments, R 1’ is an optionally substituted saturated or unsaturated linear or branched C 1 -C 20 In some embodiments, R 1’ is an optionally substituted saturated or unsaturated linear or branched C 1 -C 15 In some embodiments, R 1’ is an optionally substituted saturated or unsaturated linear or branched C 1 -C 12 In some embodiments, R 1’ is an optionally substituted saturated or unsaturated linear or branched C 1 -C 9 In some embodiments, R 1’is an optionally substituted saturated or unsaturated linear or branched C 1 -C 6 In some embodiments, R 1’ is an optionally substituted saturated or unsaturated linear or branched C 6 -C 20 In some embodiments, R 1’ is an optionally substituted saturated or unsaturated linear or branched C 9 -C 20 In some embodiments, R 1’ is an optionally substituted saturated or unsaturated linear or branched C 12 -C 20 In some embodiments, R 1’ is an optionally substituted saturated or unsaturated linear or branched C 15 -C 20 In some embodiments, R 1’ is an optionally substituted saturated or unsaturated linear or branched C 6 -C 15 In some embodiments, R 1’ is an optionally substituted saturated or unsaturated linear or branched C 8 -C 17 It is a hydrocarbon chain.

[0155] In some embodiments, R 1’ is an optionally substituted saturated or unsaturated linear or branched C 6 In some embodiments, R 1’ is an optionally substituted saturated or unsaturated linear or branched C 7 In some embodiments, R 1’ is an optionally substituted saturated or unsaturated linear or branched C 8 In some embodiments, R 1’ is an optionally substituted saturated or unsaturated linear or branched C 9 In some embodiments, R 1’is an optionally substituted saturated or unsaturated linear or branched C 10 In some embodiments, R 1’ is an optionally substituted saturated or unsaturated linear or branched C 11 In some embodiments, R 1’ is an optionally substituted saturated or unsaturated linear or branched C 12 In some embodiments, R 1’ is an optionally substituted saturated or unsaturated linear or branched C 13 In some embodiments, R 1’ is an optionally substituted saturated or unsaturated linear or branched C 14 In some embodiments, R 1’ is an optionally substituted saturated or unsaturated linear or branched C 15 In some embodiments, R 1’ is an optionally substituted saturated or unsaturated linear or branched C 16 In some embodiments, R 1’ is an optionally substituted saturated or unsaturated linear or branched C 17 In some embodiments, R 1’ is an optionally substituted saturated or unsaturated linear or branched C 18 In some embodiments, R 1’ is an optionally substituted saturated or unsaturated linear or branched C 19 In some embodiments, R 1’ is an optionally substituted saturated or unsaturated linear or branched C 20 It is a hydrocarbon chain.

[0156] In some embodiments, R 1’ is a saturated or unsaturated linear or branched C optionally substituted with 1 to 6 halogen atoms; 1 -C 20 In some embodiments, R 1’is a saturated or unsaturated linear or branched C optionally substituted with 1 to 6 halogen atoms; 1 -C 15 In some embodiments, R 1’ is a saturated or unsaturated linear or branched C optionally substituted with 1 to 6 halogen atoms; 1 -C 12 In some embodiments, R 1’ is a saturated or unsaturated linear or branched C optionally substituted with 1 to 6 halogen atoms; 1 -C 9 In some embodiments, R 1’ is a saturated or unsaturated linear or branched C optionally substituted with 1 to 6 halogen atoms; 1 -C 6 In some embodiments, R 1’ is a saturated or unsaturated linear or branched C optionally substituted with 1 to 6 halogen atoms; 6 -C 20 In some embodiments, R 1’ is a saturated or unsaturated linear or branched C optionally substituted with 1 to 6 halogen atoms; 9 -C 20 In some embodiments, R 1’ is a saturated or unsaturated linear or branched C optionally substituted with 1 to 6 halogen atoms; 12 -C 20 In some embodiments, R 1’ is a saturated or unsaturated linear or branched C optionally substituted with 1 to 6 halogen atoms; 15 -C 20 In some embodiments, R 1’ is a saturated or unsaturated linear or branched C optionally substituted with 1 to 6 halogen atoms; 6 -C 15 In some embodiments, R 1’ is a saturated or unsaturated linear or branched C optionally substituted with 1 to 6 halogen atoms; 8 -C 17It is a hydrocarbon chain.

[0157] In some embodiments, R 1’ is a saturated or unsaturated linear or branched C optionally substituted with 1 to 6 halogen atoms; 6 In some embodiments, R 1’ is a saturated or unsaturated linear or branched C optionally substituted with 1 to 6 halogen atoms; 7 In some embodiments, R 1’ is a saturated or unsaturated linear or branched C optionally substituted with 1 to 6 halogen atoms; 8 In some embodiments, R 1’ is a saturated or unsaturated linear or branched C optionally substituted with 1 to 6 halogen atoms; 9 In some embodiments, R 1’ is a saturated or unsaturated linear or branched C optionally substituted with 1 to 6 halogen atoms; 10 In some embodiments, R 1’ is a saturated or unsaturated linear or branched C optionally substituted with 1 to 6 halogen atoms; 11 In some embodiments, R 1’ is a saturated or unsaturated linear or branched C optionally substituted with 1 to 6 halogen atoms; 12 In some embodiments, R 1’ is a saturated or unsaturated linear or branched C optionally substituted with 1 to 6 halogen atoms; 13 In some embodiments, R 1’ is a saturated or unsaturated linear or branched C optionally substituted with 1 to 6 halogen atoms; 14 In some embodiments, R 1’ is a saturated or unsaturated linear or branched C optionally substituted with 1 to 6 halogen atoms; 15 In some embodiments, R 1’ is a saturated or unsaturated linear or branched C optionally substituted with 1 to 6 halogen atoms;16 In some embodiments, R 1’ is a saturated or unsaturated linear or branched C optionally substituted with 1 to 6 halogen atoms; 17 In some embodiments, R 1’ is a saturated or unsaturated linear or branched C optionally substituted with 1 to 6 halogen atoms; 18 In some embodiments, R 1’ is a saturated or unsaturated linear or branched C optionally substituted with 1 to 6 halogen atoms; 19 In some embodiments, R 1’ is a saturated or unsaturated linear or branched C optionally substituted with 1 to 6 halogen atoms; 20 It is a hydrocarbon chain.

[0158] In some embodiments, R 1’ is a saturated or unsaturated linear or branched C optionally substituted with 1 to 6 fluorine atoms; 1 -C 20 In some embodiments, R 1’ is a saturated or unsaturated linear or branched C optionally substituted with 1 to 6 fluorine atoms; 1 -C 15 In some embodiments, R 1’ is a saturated or unsaturated linear or branched C optionally substituted with 1 to 6 fluorine atoms; 1 -C 12 In some embodiments, R 1’ is a saturated or unsaturated linear or branched C optionally substituted with 1 to 6 fluorine atoms; 1 -C 9 In some embodiments, R 1’ is a saturated or unsaturated linear or branched C optionally substituted with 1 to 6 fluorine atoms; 1 -C 6 In some embodiments, R 1’ is a saturated or unsaturated linear or branched C optionally substituted with 1 to 6 fluorine atoms; 6 -C20 In some embodiments, R 1’ is a saturated or unsaturated linear or branched C optionally substituted with 1 to 6 fluorine atoms; 9 -C 20 In some embodiments, R 1’ is a saturated or unsaturated linear or branched C optionally substituted with 1 to 6 fluorine atoms; 12 -C 20 In some embodiments, R 1’ is a saturated or unsaturated linear or branched C optionally substituted with 1 to 6 fluorine atoms; 15 -C 20 In some embodiments, R 1’ is a saturated or unsaturated linear or branched C optionally substituted with 1 to 6 fluorine atoms; 6 -C 15 In some embodiments, R 1’ is a saturated or unsaturated linear or branched C optionally substituted with 1 to 6 fluorine atoms; 8 -C 17 It is a hydrocarbon chain.

[0159] In some embodiments, R 1’ is a saturated or unsaturated linear or branched C optionally substituted with 1 to 6 fluorine atoms; 6 In some embodiments, R 1’ is a saturated or unsaturated linear or branched C optionally substituted with 1 to 6 fluorine atoms; 7 In some embodiments, R 1’ is a saturated or unsaturated linear or branched C optionally substituted with 1 to 6 fluorine atoms; 8 In some embodiments, R 1’ is a saturated or unsaturated linear or branched C optionally substituted with 1 to 6 fluorine atoms; 9 In some embodiments, R 1’ is a saturated or unsaturated linear or branched C optionally substituted with 1 to 6 fluorine atoms; 10In some embodiments, R 1’ is a saturated or unsaturated linear or branched C optionally substituted with 1 to 6 fluorine atoms; 11 In some embodiments, R 1’ is a saturated or unsaturated linear or branched C optionally substituted with 1 to 6 fluorine atoms; 12 In some embodiments, R 1’ is a saturated or unsaturated linear or branched C optionally substituted with 1 to 6 fluorine atoms; 13 In some embodiments, R 1’ is a saturated or unsaturated linear or branched C optionally substituted with 1 to 6 fluorine atoms; 14 In some embodiments, R 1’ is a saturated or unsaturated linear or branched C optionally substituted with 1 to 6 fluorine atoms; 15 In some embodiments, R 1’ is a saturated or unsaturated linear or branched C optionally substituted with 1 to 6 fluorine atoms; 16 In some embodiments, R 1’ is a saturated or unsaturated linear or branched C optionally substituted with 1 to 6 fluorine atoms; 17 In some embodiments, R 1’ is a saturated or unsaturated linear or branched C optionally substituted with 1 to 6 fluorine atoms; 18 In some embodiments, R 1’ is a saturated or unsaturated linear or branched C optionally substituted with 1 to 6 fluorine atoms; 19 In some embodiments, R 1’ is a saturated or unsaturated linear or branched C optionally substituted with 1 to 6 fluorine atoms; 20 It is a hydrocarbon chain.

[0160] In some embodiments, R 1’ is a saturated or unsaturated linear or branched C substituted with 1 to 6 fluorine atoms. 1 -C20 In some embodiments, R 1’ is a saturated or unsaturated linear or branched C substituted with 1 to 6 fluorine atoms. 1 -C 15 In some embodiments, R 1’ is a saturated or unsaturated linear or branched C substituted with 1 to 6 fluorine atoms. 1 -C 12 In some embodiments, R 1’ is a saturated or unsaturated linear or branched C substituted with 1 to 6 fluorine atoms. 1 -C 9 In some embodiments, R 1’ is a saturated or unsaturated linear or branched C substituted with 1 to 6 fluorine atoms. 1 -C 6 In some embodiments, R 1’ is a saturated or unsaturated linear or branched C substituted with 1 to 6 fluorine atoms. 6 -C 20 In some embodiments, R 1’ is a saturated or unsaturated linear or branched C substituted with 1 to 6 fluorine atoms. 9 -C 20 In some embodiments, R 1’ is a saturated or unsaturated linear or branched C substituted with 1 to 6 fluorine atoms. 12 -C 20 In some embodiments, R 1’ is a saturated or unsaturated linear or branched C substituted with 1 to 6 fluorine atoms. 15 -C 20 In some embodiments, R 1’ is a saturated or unsaturated linear or branched C substituted with 1 to 6 fluorine atoms. 6 -C 15 In some embodiments, R 1’ is a saturated or unsaturated linear or branched C substituted with 1 to 6 fluorine atoms. 8 -C 17 It is a hydrocarbon chain.

[0161] In some embodiments, R 1’ is a saturated or unsaturated linear or branched C substituted with 1 to 6 fluorine atoms. 6 In some embodiments, R 1’ is a saturated or unsaturated linear or branched C substituted with 1 to 6 fluorine atoms. 7 In some embodiments, R 1’ is a saturated or unsaturated linear or branched C substituted with 1 to 6 fluorine atoms. 8 In some embodiments, R 1’ is a saturated or unsaturated linear or branched C substituted with 1 to 6 fluorine atoms. 9 In some embodiments, R 1’ is a saturated or unsaturated linear or branched C substituted with 1 to 6 fluorine atoms. 10 In some embodiments, R 1’ is a saturated or unsaturated linear or branched C substituted with 1 to 6 fluorine atoms. 11 In some embodiments, R 1’ is a saturated or unsaturated linear or branched C substituted with 1 to 6 fluorine atoms. 12 In some embodiments, R 1’ is a saturated or unsaturated linear or branched C substituted with 1 to 6 fluorine atoms. 13 In some embodiments, R 1’ is a saturated or unsaturated linear or branched C substituted with 1 to 6 fluorine atoms. 14 In some embodiments, R 1’ is a saturated or unsaturated linear or branched C substituted with 1 to 6 fluorine atoms. 15 In some embodiments, R 1’ is a saturated or unsaturated linear or branched C substituted with 1 to 6 fluorine atoms. 16 In some embodiments, R 1’ is a saturated or unsaturated linear or branched C substituted with 1 to 6 fluorine atoms. 17In some embodiments, R 1’ is a saturated or unsaturated linear or branched C substituted with 1 to 6 fluorine atoms. 18 In some embodiments, R 1’ is a saturated or unsaturated linear or branched C substituted with 1 to 6 fluorine atoms. 19 In some embodiments, R 1’ is a saturated or unsaturated linear or branched C substituted with 1 to 6 fluorine atoms. 20 It is a hydrocarbon chain.

[0162] In some embodiments, R 1’ is a saturated or unsaturated linear or branched C 1 -C 20 In some embodiments, R 1’ is a saturated or unsaturated linear or branched C 1 -C 15 In some embodiments, R 1’ is a saturated or unsaturated linear or branched C 1 -C 12 In some embodiments, R 1’ is a saturated or unsaturated linear or branched C 1 -C 9 In some embodiments, R 1’ is a saturated or unsaturated linear or branched C 1 -C 6 In some embodiments, R 1’ is a saturated or unsaturated linear or branched C 6 -C 20 In some embodiments, R 1’ is a saturated or unsaturated linear or branched C 9 -C 20 In some embodiments, R 1’ is a saturated or unsaturated linear or branched C 12 -C 20 In some embodiments, R 1’ is a saturated or unsaturated linear or branched C 15 -C 20In some embodiments, R 1’ is a saturated or unsaturated linear or branched C 6 -C 15 In some embodiments, R 1’ is a saturated or unsaturated linear or branched C 8 -C 17 It is a hydrocarbon chain.

[0163] In some embodiments, R 1’ is a saturated or unsaturated linear or branched C 6 In some embodiments, R 1’ is a saturated or unsaturated linear or branched C 7 In some embodiments, R 1’ is a saturated or unsaturated linear or branched C 8 In some embodiments, R 1’ is a saturated or unsaturated linear or branched C 9 In some embodiments, R 1’ is a saturated or unsaturated linear or branched C 10 In some embodiments, R 1’ is a saturated or unsaturated linear or branched C 11 In some embodiments, R 1’ is a saturated or unsaturated linear or branched C 12 In some embodiments, R 1’ is a saturated or unsaturated linear or branched C 13 In some embodiments, R 1’ is a saturated or unsaturated linear or branched C 14 In some embodiments, R 1’ is a saturated or unsaturated linear or branched C 15 In some embodiments, R 1’ is a saturated or unsaturated linear or branched C 16 In some embodiments, R 1’ is a saturated or unsaturated linear or branched C 17 In some embodiments, R1’ is a saturated or unsaturated linear or branched C 18 In some embodiments, R 1’ is a saturated or unsaturated linear or branched C 19 In some embodiments, R 1’ is a saturated or unsaturated linear or branched C 20 It is a hydrocarbon chain.

[0164] In some embodiments, R 1’ is an optionally substituted 3-12 membered saturated or partially unsaturated carbocyclyl. In some embodiments, R 1’ is an optionally substituted 3- to 7-membered saturated or partially unsaturated carbocyclyl. In some embodiments, R 1’ is an optionally substituted 5-6 membered saturated or partially unsaturated carbocyclyl. In some embodiments, R 1 is optionally substituted cyclopropyl. In some embodiments, R 1’ is optionally substituted cyclobutyl. In some embodiments, R 1’ is optionally substituted cyclopentyl. In some embodiments, R 1’ is optionally substituted cyclohexyl. In some embodiments, R 1’ is an optionally substituted cycloheptyl. In some embodiments, R 1’ Optionally, C 1-6 Aliphatic (e.g., C 1-6 and cyclohexyl substituted with alkyl.

[0165] In some embodiments, R 1’ is an optionally substituted 1-adamantyl. In some embodiments, R 1’ is an optionally substituted 2-adamantyl. In some embodiments, R 1’ is an optionally substituted sterolyl. In some embodiments, R 1’ is optionally substituted phenyl. In some embodiments, R 1is 1-adamantyl. In some embodiments, R 1’ is 2-adamantyl. In some embodiments, R 1’ is sterolyl. In some embodiments, R 1’ is phenyl. In some embodiments, R 1’ teeth, [ka] It is.

[0166] In some embodiments, R 1’ is selected from the group consisting of: [ka] 1-adamantyl, [ka]

[0167] In some embodiments, R 1’ is selected from the group consisting of: [ka] 1-adamantyl, [ka]

[0168] In some embodiments, R 1’ is selected from the group consisting of: [ka] 1-adamantyl, [ka]

[0169] In some embodiments, R1 and R 1’ None of the R 1 does not contain a 1,4-diene moiety. 1’ does not contain a 1,4-diene moiety.

[0170] In some embodiments of any of the formulas described herein, each L 4 are independently divalent saturated or unsaturated linear or branched C 1 -C 6 In some embodiments, each L 4 are independently divalent saturated or unsaturated linear or branched C 1 -C 5 In some embodiments, each L 4 are independently divalent saturated or unsaturated linear or branched C 1 -C 4 In some embodiments, each L 4 are independently divalent saturated or unsaturated linear or branched C 1 -C 3 In some embodiments, each L 4 are independently -CH 2 -, -CH 2 CH 2 - or -CH 2 CH 2 CH 2 In some embodiments, each L 4 -CH 2 In some embodiments, each L 4 -CH 2 CH 2 In some embodiments, each L 4 -CH 2 CH 2 CH 2 -It is.

[0171] In some embodiments of any of the formulas described herein, each A 1 and A 2 independently represents an optionally substituted C 1 -C 20Aliphatic or -L 5 -R 5 or Or A 1 and A 2 may be taken together with their intervening atoms to form an optionally substituted ring, [ka] where x and # are as described above and herein.

[0172] In some embodiments, A 1 and A 2 are the same. In some embodiments, A 1 and A 2 is different.

[0173] In some embodiments, A 1 is an optionally substituted C 1 -C 20 In some embodiments, A is aliphatic. 1 is an optionally substituted C 1 -C 15 In some embodiments, A is aliphatic. 1 is an optionally substituted C 1 -C 12 In some embodiments, A is aliphatic. 1 is an optionally substituted C 1 -C 9 In some embodiments, A is aliphatic. 1 is an optionally substituted C 1 -C 6 In some embodiments, A is aliphatic. 1 is an optionally substituted C 6 -C 20 In some embodiments, A is aliphatic. 1 is an optionally substituted C 6 -C 12 In some embodiments, A is aliphatic. 1 is an optionally substituted C 12 -C 20 In some embodiments, A is aliphatic.1 is an optionally substituted C 15 -C 20 In some embodiments, A is aliphatic. 1 is an optionally substituted C 6 In some embodiments, A is aliphatic. 1 is an optionally substituted C 7 In some embodiments, A is aliphatic. 1 is an optionally substituted C 8 In some embodiments, A is aliphatic. 1 is an optionally substituted C 9 In some embodiments, A is aliphatic. 1 is an optionally substituted C 10 In some embodiments, A is aliphatic. 1 is an optionally substituted C 11 In some embodiments, A is aliphatic. 1 is an optionally substituted C 12 In some embodiments, A is aliphatic. 1 is an optionally substituted C 13 In some embodiments, A is aliphatic. 1 is an optionally substituted C 14 In some embodiments, A is aliphatic. 1 is an optionally substituted C 15 In some embodiments, A is aliphatic. 1 is an optionally substituted C 16 It is aliphatic.

[0174] In some embodiments, A 1 is C 1 -C 20 In some embodiments, A is aliphatic. 1 is C 1 -C 15 In some embodiments, A is aliphatic. 1 is C 1 -C 12 In some embodiments, A is aliphatic. 1 is C 1 -C 9 In some embodiments, A is aliphatic.1 is C 1 -C 6 In some embodiments, A is aliphatic. 1 is C 6 -C 20 In some embodiments, A is aliphatic. 1 is C 6 -C 12 In some embodiments, A is aliphatic. 1 is C 12 -C 20 In some embodiments, A is aliphatic. 1 is C 15 -C 20 In some embodiments, A is aliphatic. 1 is C 6 In some embodiments, A is aliphatic. 1 is C 7 In some embodiments, A is aliphatic. 1 is C 8 In some embodiments, A is aliphatic. 1 is C 9 In some embodiments, A is aliphatic. 1 is C 10 In some embodiments, A is aliphatic. 1 is C 11 In some embodiments, A is aliphatic. 1 is C 12 In some embodiments, A is aliphatic. 1 is C 13 In some embodiments, A is aliphatic. 1 is C 14 In some embodiments, A is aliphatic. 1 is C 15 In some embodiments, A is aliphatic. 1 is C 16 It is aliphatic.

[0175] In some embodiments, A 1 is optionally substituted with 1 to 6 halogen atoms; 1 -C 20 In some embodiments, A is aliphatic. 1 is optionally substituted with 1 to 6 halogen atoms; 1-C 15 In some embodiments, A is aliphatic. 1 is optionally substituted with 1 to 6 halogen atoms; 1 -C 12 In some embodiments, A is aliphatic. 1 is optionally substituted with 1 to 6 halogen atoms; 1 -C 9 In some embodiments, A is aliphatic. 1 is optionally substituted with 1 to 6 halogen atoms; 1 -C 6 In some embodiments, A is aliphatic. 1 is optionally substituted with 1 to 6 halogen atoms; 6 -C 20 In some embodiments, A is aliphatic. 1 is optionally substituted with 1 to 6 halogen atoms; 6 -C 12 In some embodiments, A is aliphatic. 1 is optionally substituted with 1 to 6 halogen atoms; 12 -C 20 In some embodiments, A is aliphatic. 1 is optionally substituted with 1 to 6 halogen atoms; 15 -C 20 In some embodiments, A is aliphatic. 1 is optionally substituted with 1 to 6 halogen atoms; 6 In some embodiments, A is aliphatic. 1 is optionally substituted with 1 to 6 halogen atoms; 7 In some embodiments, A is aliphatic. 1 is optionally substituted with 1 to 6 halogen atoms; 8 In some embodiments, A is aliphatic. 1 is optionally substituted with 1 to 6 halogen atoms; 9 In some embodiments, A is aliphatic. 1 is optionally substituted with 1 to 6 halogen atoms; 10 In some embodiments, A is aliphatic. 1is optionally substituted with 1 to 6 halogen atoms; 11 In some embodiments, A is aliphatic. 1 is optionally substituted with 1 to 6 halogen atoms; 12 In some embodiments, A is aliphatic. 1 is optionally substituted with 1 to 6 halogen atoms; 13 In some embodiments, A is aliphatic. 1 is optionally substituted with 1 to 6 halogen atoms; 14 In some embodiments, A is aliphatic. 1 is optionally substituted with 1 to 6 halogen atoms; 15 In some embodiments, A is aliphatic. 1 is optionally substituted with 1 to 6 halogen atoms; 16 It is aliphatic.

[0176] In some embodiments, A 1 is optionally substituted with 1 to 6 fluorine atoms; 1 -C 20 In some embodiments, A is aliphatic. 1 is optionally substituted with 1 to 6 fluorine atoms; 1 -C 15 In some embodiments, A is aliphatic. 1 is optionally substituted with 1 to 6 fluorine atoms; 1 -C 12 In some embodiments, A is aliphatic. 1 is optionally substituted with 1 to 6 fluorine atoms; 1 -C 9 In some embodiments, A is aliphatic. 1 is optionally substituted with 1 to 6 fluorine atoms; 1 -C 6 In some embodiments, A is aliphatic. 1 is optionally substituted with 1 to 6 fluorine atoms; 6 -C 20 In some embodiments, A is aliphatic. 1 is optionally substituted with 1 to 6 fluorine atoms; 6-C 12 In some embodiments, A is aliphatic. 1 is optionally substituted with 1 to 6 fluorine atoms; 12 -C 20 In some embodiments, A is aliphatic. 1 is optionally substituted with 1 to 6 fluorine atoms; 15 -C 20 In some embodiments, A is aliphatic. 1 is optionally substituted with 1 to 6 fluorine atoms; 6 In some embodiments, A is aliphatic. 1 is optionally substituted with 1 to 6 fluorine atoms; 7 In some embodiments, A is aliphatic. 1 is optionally substituted with 1 to 6 fluorine atoms; 8 In some embodiments, A is aliphatic. 1 is optionally substituted with 1 to 6 fluorine atoms; 9 In some embodiments, A is aliphatic. 1 is optionally substituted with 1 to 6 fluorine atoms; 10 In some embodiments, A is aliphatic. 1 is optionally substituted with 1 to 6 fluorine atoms; 11 In some embodiments, A is aliphatic. 1 is optionally substituted with 1 to 6 fluorine atoms; 12 In some embodiments, A is aliphatic. 1 is optionally substituted with 1 to 6 fluorine atoms; 13 In some embodiments, A is aliphatic. 1 is optionally substituted with 1 to 6 fluorine atoms; 14 In some embodiments, A is aliphatic. 1 is optionally substituted with 1 to 6 fluorine atoms; 15 In some embodiments, A is aliphatic. 1 is optionally substituted with 1 to 6 fluorine atoms; 16 It is aliphatic.

[0177] In some embodiments, A1 is a C substituted with 1 to 6 fluorine atoms. 1 -C 20 In some embodiments, A is aliphatic. 1 is a C substituted with 1 to 6 fluorine atoms. 1 -C 15 In some embodiments, A is aliphatic. 1 is a C substituted with 1 to 6 fluorine atoms. 1 -C 12 In some embodiments, A is aliphatic. 1 is a C substituted with 1 to 6 fluorine atoms. 1 -C 9 In some embodiments, A is aliphatic. 1 is a C substituted with 1 to 6 fluorine atoms. 1 -C 6 In some embodiments, A is aliphatic. 1 is a C substituted with 1 to 6 fluorine atoms. 6 -C 20 In some embodiments, A is aliphatic. 1 is a C substituted with 1 to 6 fluorine atoms. 6 -C 12 In some embodiments, A is aliphatic. 1 is a C substituted with 1 to 6 fluorine atoms. 12 -C 20 In some embodiments, A is aliphatic. 1 is a C substituted with 1 to 6 fluorine atoms. 15 -C 20 In some embodiments, A is aliphatic. 1 is a C substituted with 1 to 6 fluorine atoms. 6 In some embodiments, A is aliphatic. 1 is a C substituted with 1 to 6 fluorine atoms. 7 In some embodiments, A is aliphatic. 1 is a C substituted with 1 to 6 fluorine atoms. 8 In some embodiments, A is aliphatic. 1 is a C substituted with 1 to 6 fluorine atoms. 9 In some embodiments, A is aliphatic. 1 is a C substituted with 1 to 6 fluorine atoms. 10In some embodiments, A is aliphatic. 1 is a C substituted with 1 to 6 fluorine atoms. 11 In some embodiments, A is aliphatic. 1 is a C substituted with 1 to 6 fluorine atoms. 12 In some embodiments, A is aliphatic. 1 is a C substituted with 1 to 6 fluorine atoms. 13 In some embodiments, A is aliphatic. 1 is a C substituted with 1 to 6 fluorine atoms. 14 In some embodiments, A is aliphatic. 1 is a C substituted with 1 to 6 fluorine atoms. 15 In some embodiments, A is aliphatic. 1 is a C substituted with 1 to 6 fluorine atoms. 16 It is aliphatic.

[0178] In some embodiments, A 1 teeth, [ka] It is.

[0179] In some embodiments, A 2 is an optionally substituted C 1 -C 20 In some embodiments, A is aliphatic. 2 is an optionally substituted C 1 -C 15 In some embodiments, A is aliphatic. 2 is an optionally substituted C 1 -C 12 In some embodiments, A is aliphatic. 2 is an optionally substituted C 1 -C 9 In some embodiments, A is aliphatic. 2 is an optionally substituted C 1 -C 6 In some embodiments, A is aliphatic. 2 is an optionally substituted C 6 -C20 In some embodiments, A is aliphatic. 2 is an optionally substituted C 6 -C 12 In some embodiments, A is aliphatic. 2 is an optionally substituted C 12 -C 20 In some embodiments, A is aliphatic. 2 is an optionally substituted C 15 -C 20 In some embodiments, A is aliphatic. 2 is an optionally substituted C 6 In some embodiments, A is aliphatic. 2 is an optionally substituted C 7 In some embodiments, A is aliphatic. 2 is an optionally substituted C 8 In some embodiments, A is aliphatic. 2 is an optionally substituted C 9 In some embodiments, A is aliphatic. 2 is an optionally substituted C 10 In some embodiments, A is aliphatic. 2 is an optionally substituted C 11 In some embodiments, A is aliphatic. 2 is an optionally substituted C 12 In some embodiments, A is aliphatic. 2 is an optionally substituted C 13 In some embodiments, A is aliphatic. 2 is an optionally substituted C 14 In some embodiments, A is aliphatic. 2 is an optionally substituted C 15 In some embodiments, A is aliphatic. 2 is an optionally substituted C 16 It is aliphatic.

[0180] In some embodiments, A 2 is C 1 -C 20 In some embodiments, A is aliphatic. 2is C 1 -C 15 In some embodiments, A is aliphatic. 2 is C 1 -C 12 In some embodiments, A is aliphatic. 2 is C 1 -C 9 In some embodiments, A is aliphatic. 2 is C 1 -C 6 In some embodiments, A is aliphatic. 2 is C 6 -C 20 In some embodiments, A is aliphatic. 2 is C 6 -C 12 In some embodiments, A is aliphatic. 2 is C 12 -C 20 In some embodiments, A is aliphatic. 2 is C 15 -C 20 In some embodiments, A is aliphatic. 2 is C 6 In some embodiments, A is aliphatic. 2 is C 7 In some embodiments, A is aliphatic. 2 is C 8 In some embodiments, A is aliphatic. 2 is C 9 In some embodiments, A is aliphatic. 2 is C 10 In some embodiments, A is aliphatic. 2 is C 11 In some embodiments, A is aliphatic. 2 is C 12 In some embodiments, A is aliphatic. 2 is C 13 In some embodiments, A is aliphatic. 2 is C 14 In some embodiments, A is aliphatic. 2 is C 15 In some embodiments, A is aliphatic. 2 is C 16 It is aliphatic.

[0181] In some embodiments, A 2 is optionally substituted with 1 to 6 halogen atoms; 1 -C 20 In some embodiments, A is aliphatic. 2 is optionally substituted with 1 to 6 halogen atoms; 1 -C 15 In some embodiments, A is aliphatic. 2 is optionally substituted with 1 to 6 halogen atoms; 1 -C 12 In some embodiments, A is aliphatic. 2 is optionally substituted with 1 to 6 halogen atoms; 1 -C 9 In some embodiments, A is aliphatic. 2 is optionally substituted with 1 to 6 halogen atoms; 1 -C 6 In some embodiments, A is aliphatic. 2 is optionally substituted with 1 to 6 halogen atoms; 6 -C 20 In some embodiments, A is aliphatic. 2 is optionally substituted with 1 to 6 halogen atoms; 6 -C 12 In some embodiments, A is aliphatic. 2 is optionally substituted with 1 to 6 halogen atoms; 12 -C 20 In some embodiments, A is aliphatic. 2 is optionally substituted with 1 to 6 halogen atoms; 15 -C 20 In some embodiments, A is aliphatic. 2 is optionally substituted with 1 to 6 halogen atoms; 6 In some embodiments, A is aliphatic. 2 is optionally substituted with 1 to 6 halogen atoms; 7 In some embodiments, A is aliphatic. 2 is optionally substituted with 1 to 6 halogen atoms; 8In some embodiments, A is aliphatic. 2 is optionally substituted with 1 to 6 halogen atoms; 9 In some embodiments, A is aliphatic. 2 is optionally substituted with 1 to 6 halogen atoms; 10 In some embodiments, A is aliphatic. 2 is optionally substituted with 1 to 6 halogen atoms; 11 In some embodiments, A is aliphatic. 2 is optionally substituted with 1 to 6 halogen atoms; 12 In some embodiments, A is aliphatic. 2 is optionally substituted with 1 to 6 halogen atoms; 13 In some embodiments, A is aliphatic. 2 is optionally substituted with 1 to 6 halogen atoms; 14 In some embodiments, A is aliphatic. 2 is optionally substituted with 1 to 6 halogen atoms; 15 In some embodiments, A is aliphatic. 2 is optionally substituted with 1 to 6 halogen atoms; 16 It is aliphatic.

[0182] In some embodiments, A 2 is optionally substituted with 1 to 6 fluorine atoms; 1 -C 20 In some embodiments, A is aliphatic. 2 is optionally substituted with 1 to 6 fluorine atoms; 1 -C 15 In some embodiments, A is aliphatic. 2 is optionally substituted with 1 to 6 fluorine atoms; 1 -C 12 In some embodiments, A is aliphatic. 2 is optionally substituted with 1 to 6 fluorine atoms; 1 -C 9 In some embodiments, A is aliphatic. 2 is optionally substituted with 1 to 6 fluorine atoms;1 -C 6 In some embodiments, A is aliphatic. 2 is optionally substituted with 1 to 6 fluorine atoms; 6 -C 20 In some embodiments, A is aliphatic. 2 is optionally substituted with 1 to 6 fluorine atoms; 6 -C 12 In some embodiments, A is aliphatic. 2 is optionally substituted with 1 to 6 fluorine atoms; 12 -C 20 In some embodiments, A is aliphatic. 2 is optionally substituted with 1 to 6 fluorine atoms; 15 -C 20 In some embodiments, A is aliphatic. 1 is optionally substituted with 1 to 6 fluorine atoms; 6 In some embodiments, A is aliphatic. 2 is optionally substituted with 1 to 6 fluorine atoms; 7 In some embodiments, A is aliphatic. 2 is optionally substituted with 1 to 6 fluorine atoms; 8 In some embodiments, A is aliphatic. 2 is optionally substituted with 1 to 6 fluorine atoms; 9 In some embodiments, A is aliphatic. 2 is optionally substituted with 1 to 6 fluorine atoms; 10 In some embodiments, A is aliphatic. 2 is optionally substituted with 1 to 6 fluorine atoms; 11 In some embodiments, A is aliphatic. 2 is optionally substituted with 1 to 6 fluorine atoms; 12 In some embodiments, A is aliphatic. 2 is optionally substituted with 1 to 6 fluorine atoms; 13 In some embodiments, A is aliphatic. 2 is optionally substituted with 1 to 6 fluorine atoms; 14 In some embodiments, A is aliphatic.2 is optionally substituted with 1 to 6 fluorine atoms; 15 In some embodiments, A is aliphatic. 2 is optionally substituted with 1 to 6 fluorine atoms; 16 It is aliphatic.

[0183] In some embodiments, A 2 is a C substituted with 1 to 6 fluorine atoms. 1 -C 20 In some embodiments, A is aliphatic. 2 is a C substituted with 1 to 6 fluorine atoms. 1 -C 15 In some embodiments, A is aliphatic. 2 is a C substituted with 1 to 6 fluorine atoms. 1 -C 12 In some embodiments, A is aliphatic. 2 is a C substituted with 1 to 6 fluorine atoms. 1 -C 9 In some embodiments, A is aliphatic. 2 is a C substituted with 1 to 6 fluorine atoms. 1 -C 6 In some embodiments, A is aliphatic. 2 is a C substituted with 1 to 6 fluorine atoms. 6 -C 20 In some embodiments, A is aliphatic. 2 is a C substituted with 1 to 6 fluorine atoms. 6 -C 12 In some embodiments, A is aliphatic. 2 is a C substituted with 1 to 6 fluorine atoms. 12 -C 20 In some embodiments, A is aliphatic. 2 is a C substituted with 1 to 6 fluorine atoms. 15 -C 20 In some embodiments, A is aliphatic. 2 is a C substituted with 1 to 6 fluorine atoms. 6 In some embodiments, A is aliphatic. 2 is a C substituted with 1 to 6 fluorine atoms. 7In some embodiments, A is aliphatic. 2 is a C substituted with 1 to 6 fluorine atoms. 8 In some embodiments, A is aliphatic. 2 is a C substituted with 1 to 6 fluorine atoms. 9 In some embodiments, A is aliphatic. 2 is a C substituted with 1 to 6 fluorine atoms. 10 In some embodiments, A is aliphatic. 2 is a C substituted with 1 to 6 fluorine atoms. 11 In some embodiments, A is aliphatic. 2 is a C substituted with 1 to 6 fluorine atoms. 12 In some embodiments, A is aliphatic. 2 is a C substituted with 1 to 6 fluorine atoms. 13 In some embodiments, A is aliphatic. 2 is a C substituted with 1 to 6 fluorine atoms. 14 In some embodiments, A is aliphatic. 2 is a C substituted with 1 to 6 fluorine atoms. 15 In some embodiments, A is aliphatic. 2 is a C substituted with 1 to 6 fluorine atoms. 16 It is aliphatic.

[0184] In some embodiments, A 2 teeth, [ka] It is.

[0185] In some embodiments, A 1 -L 5 -R 5 In some embodiments, A 2 -L 5 -R 5 It is.

[0186] In some embodiments, A 1 and A 2 may be taken together with their intervening atoms to form an optionally substituted ring, [ka] where x and # are as described above and herein.

[0187] In some embodiments, A 1 and A 2 may be taken together with their intervening atoms to form an optionally substituted ring selected from the group consisting of: [ka]

[0188] In some embodiments, A 1 and A 2 may be taken together with their intervening atoms to form a ring selected from the group consisting of: [ka]

[0189] In some embodiments of any of the formulas described herein, each L 5 are independently divalent saturated or unsaturated linear or branched C 1 -C 20 In some embodiments, each L is a hydrocarbon chain in which one to three methylene units are optionally and independently replaced with -O- or -NR-. 5 independently, divalent saturated or unsaturated linear or branched C 1 -C 12 In some embodiments, each L is a hydrocarbon chain in which one to three methylene units are optionally and independently replaced with -O- or -NR-. 5 independently, divalent saturated or unsaturated linear or branched C 12 -C 20 A hydrocarbon chain in which one to three methylene units are optionally and independently replaced by -O- or -NR-.

[0190] In some embodiments of any of the formulas described herein, each L 5 are independently divalent saturated or unsaturated linear or branched C 1 -C 20 In some embodiments, each L 5 are independently divalent saturated or unsaturated linear or branched C 1 -C 12 In some embodiments, each L 5 are independently divalent saturated or unsaturated linear or branched C 12 -C 20 It is a hydrocarbon chain.

[0191] In some embodiments of any of the formulas described herein, each R 5 is independently an optionally substituted group selected from 6-10 membered aryl or saturated or partially unsaturated 3-8 membered carbocyclyl. 5 is independently an optionally substituted 6-10 membered aryl (e.g., phenyl). In some embodiments, each R 5 is independently an optionally substituted saturated or partially unsaturated 3-8 membered carbocyclyl (eg, cyclohexyl).

[0192] In some embodiments of any of the formulas described herein, X 1 is a covalent bond, -O-, or -NR-. 1 is a covalent bond. In some embodiments, X 1 In some embodiments, X is -O-. 1 In some embodiments, X is -NR-. 1 is -NH-.

[0193] In some embodiments of any of the formulas described herein, X 2 is a covalent bond or an optionally substituted divalent saturated or unsaturated linear or branched C 1 -C 12In some embodiments, X is a hydrocarbon chain in which one to three methylene units are optionally and independently replaced with -O- or -NR-. 2 is a covalent bond. In some embodiments, X 2 is an optionally substituted divalent saturated or unsaturated linear or branched C 1 -C 12 A hydrocarbon chain in which one to three methylene units are optionally and independently replaced by -O- or -NR-.

[0194] In some embodiments, X 2 is an optionally substituted divalent saturated or unsaturated linear or branched C 1 -C 9 In some embodiments, X is a hydrocarbon chain in which one to three methylene units are optionally and independently replaced with -O- or -NR-. 2 is an optionally substituted divalent saturated or unsaturated linear or branched C 1 -C 6 In some embodiments, X is a hydrocarbon chain in which one to three methylene units are optionally and independently replaced with -O- or -NR-. 2 is an optionally substituted divalent saturated or unsaturated linear or branched C 1 -C 4 In some embodiments, X is a hydrocarbon chain in which one to three methylene units are optionally and independently replaced with -O- or -NR-. 2 is an optionally substituted divalent saturated or unsaturated linear or branched C 1 -C 3 A hydrocarbon chain in which one to three methylene units are optionally and independently replaced by -O- or -NR-.

[0195] In some embodiments, X 2 is an optionally substituted divalent saturated or unsaturated linear or branched C 1 -C 12In some embodiments, X is a hydrocarbon chain, in which one methylene unit is optionally and independently replaced with -O- or -NR-. 2 is an optionally substituted divalent saturated or unsaturated linear or branched C 1 -C 12 In some embodiments, X is a hydrocarbon chain in which one methylene unit is replaced with -O- or -NR-. 2 is an optionally substituted divalent saturated or unsaturated linear or branched C 1 -C 9 In some embodiments, X is a hydrocarbon chain in which one methylene unit is replaced with -O- or -NR-. 2 is an optionally substituted divalent saturated or unsaturated linear or branched C 1 -C 6 In some embodiments, X is a hydrocarbon chain in which one methylene unit is replaced with -O- or -NR-. 2 is an optionally substituted divalent saturated or unsaturated linear or branched C 1 -C 3 It is a hydrocarbon chain in which one methylene unit is replaced by -O- or -NR-.

[0196] In some embodiments, X 2 is an optionally substituted divalent saturated or unsaturated linear or branched C 1 -C 12 In some embodiments, X 2 is an optionally substituted divalent saturated or unsaturated linear or branched C 1 -C 9 In some embodiments, X 2 is an optionally substituted divalent saturated or unsaturated linear or branched C 1 -C 6 In some embodiments, X 2 is an optionally substituted divalent saturated or unsaturated linear or branched C 1 -C 3 In some embodiments, X 2 is an optionally substituted -CH2 -, -CH 2 CH 2 -, -CH 2 CH 2 CH 2 -It is.

[0197] In some embodiments, X 2 is a divalent saturated or unsaturated linear or branched C 1 -C 12 In some embodiments, X 2 is a divalent saturated or unsaturated linear or branched C 1 -C 9 In some embodiments, X 2 is a divalent saturated or unsaturated linear or branched C 1 -C 6 In some embodiments, X 2 is a divalent saturated or unsaturated linear or branched C 1 -C 3 In some embodiments, X 2 -CH 2 -, -CH 2 CH 2 -, -CH 2 CH 2 CH 2 In some embodiments, X 2 -CH 2 In some embodiments, X 2 -CH 2 CH 2 In some embodiments, X 2 -CH 2 CH 2 CH 2 -It is.

[0198] In some embodiments of any of the formulas described herein, X 3 is hydrogen or -Cy B In some embodiments, X 3 is hydrogen. In some embodiments, X 3 -Cy B In some embodiments, X 3 is hydrogen, R1 or R 1’ At least one of the [ka] It is.

[0199] In some embodiments of any of the formulas described herein, Cy B is an optionally substituted ring selected from 3- to 7-membered saturated or partially unsaturated carbocyclyl; phenyl; 3- to 7-membered saturated or partially unsaturated heterocyclyl having 1-3 heteroatoms independently selected from nitrogen, oxygen, and sulfur; and 5- to 6-membered heteroaryl having 1-3 heteroatoms independently selected from nitrogen, oxygen, and sulfur.

[0200] In some embodiments, Cy B is an optionally substituted 3- to 7-membered saturated or partially unsaturated carbocyclyl. B is optionally substituted phenyl. In some embodiments, Cy B is an optionally substituted 3-7 membered saturated or partially unsaturated heterocyclyl having 1-3 heteroatoms independently selected from nitrogen, oxygen, and sulfur. B is an optionally substituted 5-6 membered heteroaryl having 1-3 heteroatoms independently selected from nitrogen, oxygen, and sulfur.

[0201] In some embodiments, Cy B is an optionally substituted ring selected from cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, and cycloheptyl. B is selected from cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, and cycloheptyl.

[0202] In some embodiments, Cy Bis an optionally substituted ring selected from aziridinyl, azetidinyl, pyrrolidinyl, and piperidinyl. B is an optionally substituted ring selected from pyrrolidinyl and piperidinyl. B is an optionally substituted pyrrolidinyl. In some embodiments, Cy B is optionally substituted piperidinyl.

[0203] In some embodiments, Cy B Optionally, C 1 -C 6 In some embodiments, Cy is selected from aliphatically substituted aziridinyl, azetidinyl, pyrrolidinyl, and piperidinyl. B Optionally, C 1 -C 6 In some embodiments, Cy is selected from aliphatically substituted pyrrolidinyl and piperidinyl. B Optionally, C 1 -C 6 In some embodiments, Cy is an aliphatically substituted pyrrolidinyl. B Optionally, C 1 -C 6 It is an aliphatically substituted piperidinyl.

[0204] In some embodiments, Cy B Optionally, C 1 -C 3 In some embodiments, Cy is selected from aliphatically substituted aziridinyl, azetidinyl, pyrrolidinyl, and piperidinyl. B Optionally, C 1 -C 3 In some embodiments, Cy is selected from aliphatically substituted pyrrolidinyl and piperidinyl. B Optionally, C 1 -C 3 In some embodiments, Cy is an aliphatically substituted pyrrolidinyl. B Optionally, C 1 -C3 It is an aliphatically substituted piperidinyl.

[0205] In some embodiments, Cy B is selected from aziridinyl, azetidinyl, pyrrolidinyl, and piperidinyl. B is selected from pyrrolidinyl and piperidinyl. B is pyrrolidinyl. In some embodiments, Cy B is piperidinyl. B is methylpyrrolidinyl. In some embodiments, Cy B is methylpiperidinyl. In some embodiments, Cy B is ethylpyrrolidinyl. In some embodiments, Cy B is ethylpiperidinyl.

[0206] In some embodiments, Cy B is optionally substituted pyrrolidinyl or piperidinyl; Cy B The carbon atoms in X 2 In some embodiments, Cy B is optionally substituted pyrrolidinyl; Cy B The carbon atoms in X 2 In some embodiments, Cy B is optionally substituted piperidinyl; Cy B The carbon atoms in X 2 is bound to.

[0207] In some embodiments, Cy B is optionally substituted pyrrolidinyl or piperidinyl; Cy B The nitrogen atom in X 2 In some embodiments, Cy B is optionally substituted pyrrolidinyl; Cy B The nitrogen atom in X 2 In some embodiments, Cy Bis optionally substituted piperidinyl; Cy B The nitrogen atom in X 2 is bound to.

[0208] In some embodiments, Cy B is an optionally substituted ring selected from: [ka]

[0209] In some embodiments, Cy B is an optionally substituted ring selected from: [ka]

[0210] In some embodiments, Cy B teeth, [ka] and optionally a ring selected from C 1 -C 6 Aliphatically substituted.

[0211] In some embodiments, Cy B teeth, [ka] and optionally C 1 -C 6 Aliphatically substituted.

[0212] In some embodiments, Cy B teeth, [ka] and optionally substituted with methyl or ethyl.

[0213] In some embodiments, Cy B teeth, [ka] and optionally substituted with methyl or ethyl.

[0214] In some embodiments, Cy B is selected from: [ka]

[0215] In some embodiments, Cy B teeth, [ka] In some embodiments, Cy B teeth, [ka] It is.

[0216] In some embodiments of any of the formulas described herein, each R is independently hydrogen or an optionally substituted C 1 -C 6 In some embodiments, each R is an aliphatic group. In some embodiments, each R is hydrogen. In some embodiments, each R is independently an optionally substituted C 1 -C 6 In some embodiments, each R is independently an optionally substituted C 1 -C 6 In some embodiments, each R is independently an optionally substituted C 1 -C 4 In some embodiments, each R is independently an optionally substituted C alkyl group. 1 -C 2 In some embodiments, each R is independently C 1-C 6 In some embodiments, each R is independently C 1 -C 4 In some embodiments, each R is independently C 1 -C 2 In some embodiments, R is an alkyl group. In some embodiments, R is methyl. In some embodiments, R is ethyl.

[0217] In some embodiments, the compound or a pharma- ceutically acceptable salt thereof is selected from those listed in Table 1. [Table 1] TIFF2024503441000074.tif236162TIFF2024503441000075.tif241167TIFF2024503441000076.tif232162TIFF2024503441000077.tif248166 TIFF2024503441000078.tif237163TIFF2024503441000079.tif239163TIFF2024503441000080.tif247161TIFF2024503441000081.tif187168

[0218] For the purposes of this invention, chemical elements are defined as defined in the Periodic Table of the Elements, CAS version, Handbook of Chemistry and Physics, 75 th In addition, general principles of organic chemistry are described in "Organic Chemistry", Thomas Sorrell, University Science Books, Sausalito: 1999, and "March's Advanced Organic Chemistry", 5th Ed., the entire contents of which are incorporated herein by reference. th Ed., Ed.: Smith, M. B. and March, J., John Wiley & Sons, New York: 2001.

[0219] Unless otherwise stated, structures depicted herein are also meant to include all isomeric (e.g., enantiomeric, diastereomeric, and geometric (or conformational)) forms of the structure, such as the R and S configurations of each asymmetric center, Z and E double bond isomers, and Z and E stereoisomers. Thus, single stereochemical isomers as well as enantiomeric, diastereomeric, and geometric (or conformational) mixtures of the present compounds are within the scope of the present invention. Unless otherwise stated, all tautomeric forms of the compounds of the present invention are within the scope of the present invention. In addition, unless otherwise stated, structures depicted herein are also intended to include compounds that differ only in the presence of one or more isotopically enriched atoms. For example, the replacement of hydrogen with deuterium or tritium, or 13 C or 14 Within the scope of this invention are compounds having this structure that include a carbon replacement with a C-enriched carbon. Such compounds are useful, for example, as analytical tools, as probes in biological assays, or as therapeutic agents according to the invention.

[0220] Unless otherwise specified or prohibited by any of the preceding definitions of formulas I, II, III, IV-A, IV-B, VA, VB, VI-A, VI-B, VII-A, VII-B, VIII-A, VIII-B, VIII-C, VIII-D, IX-A, IX-B, IX-C, and IX-D, the variables A, B, C, D, E ... 1 , A 2 , L 1 , L 1’ , L 2 , L 2’ , L 3 , L 3’ , L 4 , L 5 , R, R 1 , R 1’ , R 5 , X 1 , X 2 , X 3 , Cy A , Cy BIt will be understood that the embodiments of, m, and x, both alone and in combination, apply to compounds of any of formulas I, II, III, IV-A, IV-B, VA, VB, VI-A, VI-B, VII-A, VII-B, VIII-A, VIII-B, VIII-C, VIII-D, IX-A, IX-B, IX-C, and IX-D.

[0221] In some embodiments, provided compounds are provided and / or utilized in a salt form (e.g., a pharma- ceutically acceptable salt form). Reference to a compound provided herein is understood to include a reference to a salt thereof, unless otherwise indicated.

[0222] It will be understood that throughout this disclosure, unless otherwise indicated, references to compounds of formula I are intended to also include species of compounds of formulae II, III, IV-A, IV-B, VA, VB, VI-A, VI-B, VII-A, VII-B, VIII-A, VIII-B, VIII-C, VIII-D, IX-A, IX-B, IX-C, and IX-D, and compounds of such formulae disclosed herein.

[0223] In some embodiments, the present disclosure encompasses the recognition that the provided compounds exhibit certain desirable characteristics, e.g., compared to a reference compound or other known compounds. For example, in some embodiments, the provided compounds exhibit more potent delivery to various cell types in one or more experiments described herein, and / or have one or more other characteristics that make them more suitable for delivery of cargo, such as therapeutic or prophylactic agents, than other known compounds. Without wishing to be bound by any particular theory, the present disclosure encompasses the recognition that the provided compounds, characterized by including a reduced number of stereocenters, exhibit certain more desirable characteristics (e.g., more potent delivery to various cell types and / or easier manufacture in one or more experiments described herein) than corresponding compounds having one or more stereocenters.

[0224] B. Ionizable lipids Among other things, the present disclosure describes compositions, preparations, nanoparticles, and / or nanomaterials that include one or more ionizable lipids described herein.

[0225] In particular, it has been surprisingly found that different ratios of ionizable lipids affect one or more functional activities, such as the desired tropism, stabilization, and drug delivery efficacy, of the compositions, preparations, nanoparticles, and / or nanomaterials described herein. For example, the present disclosure presents the surprising discovery that amounts of ionizable lipids different from those described in the art (see, e.g., U.S. Pat. No. 8,058,069 B2, or, e.g., U.S. Pat. No. 9,364,435, the contents of both of which are incorporated herein by reference in their entirety) are important and / or affect one or more functional activities of the compositions, preparations, nanoparticles, and / or nanomaterials described herein. For example, in some embodiments, compositions, preparations, nanoparticles, and / or nanomaterials having an ionizable lipid that is about 50 mol percent or less, based on the total moles of the lipid nanoparticle constituents, have been found to be useful and / or important for the functional activities of the lipid nanoparticles, such as the desired tropism, stabilization, and drug delivery efficacy, as described herein.

[0226] In some embodiments, the ionizable lipid may include an amine-containing group on the head group. In some embodiments, the ionizable lipid is or includes a compound described herein (e.g., a compound of formula I, II, III, IV-A, IV-B, VA, VB, VI-A, VI-B, VII-A, VII-B, VIII-A, VIII-B, VIII-C, VIII-D, IX-A, IX-B, IX-C, or IX-D). In some embodiments, the ionizable lipid is present in the lipid nanoparticle (LNP) preparation at about 30 mol percent to about 70 mol percent based on the total moles of the lipid nanoparticle components. In some embodiments, the ionizable lipid is present at about 33 mol percent to about 60 mol percent based on the total moles of the lipid nanoparticle components. In some embodiments, the ionizable lipid is present at about 34 mol percent to about 55 mol percent based on the total moles of the lipid nanoparticle components. In some embodiments, the ionizable lipid is present at about 33 mol percent to about 51 mol percent based on the total moles of the lipid nanoparticle components. In some embodiments, the ionizable lipid is present at about 34.7 mole percent based on the total moles of the lipid nanoparticle components. In some embodiments, the ionizable lipid is present at about 47.5 mole percent based on the total moles of the lipid nanoparticle components. In some embodiments, the ionizable lipid is present at about 50 mole percent based on the total moles of the lipid nanoparticle components.

[0227] In particular, in some embodiments, the lipid nanoparticle composition comprises an ionizable lipid. In some embodiments, the lipid nanoparticle preparation comprises an ionizable lipid, a phospholipid, a conjugate-linker lipid, and cholesterol. In some embodiments, the ionizable lipid is or comprises a compound described herein (e.g., Formula I, II, III, IV-A, IV-B, VA, VB, VI-A, VI-B, VII-A, VII-B, VIII-A, VIII-B, VIII-C, VIII-D, IX-A, IX-B, IX-C, or IX-D). In some embodiments, the ionizable lipid is present in the LNP preparation at about 30 mole percent to about 70 mole percent, based on the total moles of the lipid nanoparticle components.

[0228] C. Sterols Among other things, the present disclosure describes compositions, preparations, nanoparticles, and / or nanomaterials that include one or more sterols described herein.

[0229] In some embodiments, the sterol is cholesterol, or a variant or derivative thereof. In some embodiments, the cholesterol is modified. In some embodiments, the cholesterol is oxidized cholesterol. In some embodiments, the cholesterol is esterified cholesterol. Unmodified cholesterol may be enzymatically acted upon to form variants that are oxidized side chains or rings. In some embodiments, cholesterol may be oxidized on the beta-ring structure or on the hydrocarbon tail structure. In some embodiments, the sterol is a phytosterol. Exemplary sterols contemplated for use in the lipid nanoparticles of the present disclosure include, but are not limited to, 25-hydroxycholesterol (25-OH), 20α-hydroxycholesterol (20α-OH), 27-hydroxycholesterol, 6-keto-5α-hydroxycholesterol, 7-ketocholesterol, 7β-hydroxycholesterol, 7α-hydroxycholesterol, 7β-25-dihydroxycholesterol, beta-sitosterol, stigmasterol, brassicasterol, campesterol, or combinations thereof. In some embodiments, side chain oxidized cholesterol may enhance cargo delivery compared to other cholesterol variants. In some embodiments, the cholesterol is unmodified cholesterol.

[0230] In some embodiments, the LNP composition comprises about 20 mol percent to about 50 mol percent sterol. In some embodiments, the LNP composition comprises about 38 mol percent sterol. In some embodiments, the LNP composition comprises about 38.5 mol percent sterol. In some embodiments, the LNP composition comprises about 33.8 mol percent cholesterol. In some embodiments, the LNP composition comprises about 40 mol percent cholesterol.

[0231] D. Conjugate-Linker Lipid Among other things, the present disclosure describes compositions, preparations, nanoparticles, and / or nanomaterials that include one or more conjugate-linker lipids described herein.

[0232] In some embodiments, the conjugate-linker lipid is or includes a polyethylene glycol (PEG) lipid or a PEG-modified lipid. In some embodiments, the PEG or PEG-modified lipid may alternatively be referred to as a PEGylated lipid or a PEG lipid. The inclusion of a PEGylated lipid may be used to enhance lipid nanoparticle colloidal stability in vitro and circulation time in vivo. In some embodiments, the PEGylation is reversible in that the PEG moiety is gradually released into the blood circulation. Exemplary PEG lipids include, but are not limited to, PEG conjugated to a saturated or unsaturated alkyl chain having a length of C6-C20. PEG-modified phosphatidylethanolamine, PEG-modified phosphatidic acid, PEG-modified ceramide (PEG-CER), PEG-modified dialkylamine, PEG-modified diacylglycerol (PEG-DAG), PEG-modified dialkylglycerol, and mixtures thereof. For example, in some embodiments, the PEG lipid may be a PEG-c-DOMG, PEG-DMG, PEG-DLPE, PEG-DMPE, PEG-DPPE, PEG-DSG, or PEG-DSPE lipid.

[0233] In some embodiments, the conjugate-linker lipid comprises a polyethylene glycol lipid. In some embodiments, the conjugate-linker lipid comprises dimyristylglycerol (DMG), 1,2-dipalmitoyl-rac-glycerol, methoxypolyethylene glycol (DPG-PEG), or 1,2-distearoyl-rac-glycero-3-methylpolyoxyethylene (DSG-PEG). In some embodiments, the conjugate-linker lipid has an average molecular weight of about 500 Da to about 5000 Da. In some embodiments, the conjugate-linker lipid has an average molecular weight of about 2000 Da. In some embodiments, the LNP composition comprises about 0 mol percent to about 5 mol percent of the conjugate-linker lipid. In some embodiments, the LNP composition comprises about 1.5 mol percent of the conjugate-linker lipid. In some embodiments, the LNP composition comprises about 2.5 mol percent of the conjugate-linker lipid. In some embodiments, the LNP composition comprises about 3 mol percent of the conjugate-linker lipid.

[0234] E. Phospholipids In particular, the present disclosure describes compositions, preparations, nanoparticles, and / or nanomaterials comprising one or more phospholipids described herein. In some embodiments, the present disclosure describes compositions, preparations, nanoparticles, and / or nanomaterials comprising one or more (poly)unsaturated lipids.

[0235] In some embodiments, one or more phospholipids may be assembled into one or more lipid bilayers. In some embodiments, one or more phospholipids may include a phospholipid moiety. In some embodiments, one or more phospholipids may include one or more fatty acid moieties. In some embodiments, one or more phospholipids may include a phospholipid moiety and one or more fatty acid moieties. In some embodiments, the phospholipid moiety includes, but is not limited to, phosphatidylcholine, phosphatidylethanolamine, phosphatidylglycerol, phosphatidylserine, phosphatidic acid, 2-lysophosphatidylcholine, and sphingomyelin. In some embodiments, the fatty acid moiety includes, but is not limited to, lauric acid, myristic acid, myristoleic acid, palmitic acid, palmitoleic acid, stearic acid, oleic acid, linoleic acid, alpha linolenic acid, erucic acid, phytanic acid, arachidic acid, arachidonic acid, eicosapentaenoic acid, behenic acid, docosapentaenoic acid, and docosahexaenoic acid. Also contemplated are non-natural species, including natural species with modifications and substitutions, including branching, oxidation, cyclization, and alkynes. For example, phospholipids can be functionalized or crosslinked with one or more alkynes (e.g., alkenyl groups with one or more double bonds replaced with triple bonds). Under appropriate reaction conditions, alkyne groups can undergo copper-catalyzed cycloaddition upon exposure to azide. Such reactions can be useful for functionalizing the lipid bilayer of nanoparticle compositions to facilitate membrane permeation or cell recognition, or for conjugating nanoparticle compositions to useful components, such as targeting or imaging moieties (e.g., dyes).

[0236] Exemplary phospholipids include, but are not limited to, 1,2-distearoyl-sn-glycero-3-phosphocholine (DSPC), 1,2-dioleoyl-sn-glycero-3-phosphoethanolamine (DOPE), 1,2-dilinoleoyl-sn-glycero-3-phosphocholine (DLPC), 1,2-dimyristoyl-sn-glycerophosphocholine (DMPC), 1,2-dioleoyl-sn-glycero-3-phosphocholine (DOPC), 1,2-dipalmitoyl-sn-glycero l-3-phosphocholine (DPPC), 1,2-diundecanoyl-sn-glycerophosphocholine (DUPC), l-palmitoyl-2-oleoyl-sn-glycero-3-phosphocholine (POPC), 1,2-di-O-octadecenyl-sn-glycero-3-phosphocholine (18:0 diether PC), 1-oleoyl-2-cholesterylhemisuccinoyl l-sn-glycero-3-phosphocholine (OChemsPC), 1-hexadecyl sn-glycero-3-phosphocholine (C16 Lyso PC), 1,2-dilinolenoyl-sn-glycero-3-phosphocholine, 1,2-diarachidonoyl-sn-glycero-3-phosphocholine, 1,2-didocosahexaenoyl-sn-glycero-3-phosphocholine, 1,2-diphytanoyl-sn-glycero-3-phosphoethanolamine (ME16.0PE), 1,2-distearoyl-sn-glycero-3-phosphoethanolamine, 1,2-dilinoleoyl-sn-glycero-3-phosphoethanolamine, 1,2-dilinolenoyl-sn-glycero-3-phosphoethanolamine, 1,2-diarachidonoyl-sn-glycero-3-phosphoethanolamine, 1,2-didocosahexaenoyl-sn-glycero-3-phosphoethanolamine, 1,2-dioleoyl-sn-glycero-3-phospho-rac-(1-glycerol) sodium salt (DOPG), dipalmitoyl phosphatidylglycerol (DPPG), palmitoyl oleoyl phosphatidylethanolamine (POPE), distearoyl In some embodiments, the phospholipid may be 1-stearoyl-2-oleoyl-phosphatidylethanolamine (DSPE), dipalmitoylphosphatidylethanolamine (DPPE), dimyristoylphosphoethanolamine (DMPE), 1-stearoyl-2-oleoyl-phosphatidylethanolamine (SOPE), 1-stearoyl-2-oleoylphosphatidylcholine (SOPC), sphingomyelin, phosphatidylcholine, phosphatidylethanolamine, phosphatidylserine, phosphatidylinositol, phosphatidic acid, palmitoyloleoylphosphatidylcholine, lysophosphatidylcholine, lysophosphatidylethanolamine (LPE), or a combination thereof. In some embodiments, the phospholipid is DSPC. In some embodiments, the phospholipid is DMPC.

[0237] In some embodiments, the phospholipid comprises 1,2-dioleoyl-sn-glycero-3-phosphoethanolamine-N-(succinyl) (succinyl PE), 1,2-distearoyl-sn-glycero-3-phosphocholine (DSPC), cholesterol, 1,2-distearoyl-sn-glycero-3-phosphoethanolamine (DSPE), 1,2-dipalmitoyl-sn-glycero-3-phosphoethanolamine-N-(succinyl) (succinyl-DPPE), 1,2-dioleoyl-sn-glycero-3-phosphoethanolamine (DOPE), 1,2-dimyristoyl-sn-glycero-3-phosphocholine (DMPC), 1,2-dipalmitoyl-sn-glycero-3-phosphocholine (DPPC), or a combination thereof.

[0238] In some embodiments, the LNP composition comprises about 0 mol percent to about 15 mol percent phospholipids. In some embodiments, the LNP composition comprises about 9 mol percent phospholipids. In some embodiments, the LNP composition comprises about 10 mol percent phospholipids.

[0239] F.Diameter Among other things, the present disclosure describes compositions, preparations, nanoparticles, and / or nanomaterials having an average hydrodynamic diameter of about 30 to about 220 nm. In some embodiments, the compositions, preparations, nanoparticles, and / or nanomaterials described herein have an average hydrodynamic diameter that is about 30 nm, 35 nm, 40 nm, 45 nm, 50 nm, 55 nm, 60 nm, 65 nm, 70 nm, 75 nm, 80 nm, 85 nm, 90 nm, 95 nm, 100 nm, 105 nm, 110 nm, 115 nm, 120 nm, 125 nm, 130 nm, 135 nm, 140 nm, 145 nm, 150 nm, 155 nm, 160 nm, 165 nm, 170 nm, 175 nm, 180 nm, 185 nm, 190 nm, 195 nm, 200 nm, 205 nm, 210 nm, 215 nm, 220 nm, or any range having endpoints defined by any two of the aforesaid values. For example, in some embodiments, the compositions, preparations, nanoparticles, and / or nanomaterials described herein have an average hydrodynamic diameter of between 50 nm and 200 nm.

[0240] In some embodiments, the lipid nanoparticles described herein may have a mean hydrodynamic diameter of about 30 to about 220 nm. In some embodiments, the lipid nanoparticles described herein may have a mean hydrodynamic diameter of about 30 nm, 35 nm, 40 nm, 45 nm, 50 nm, 55 nm, 60 nm, 65 nm, 70 nm, 75 nm, 80 nm, 85 nm, 90 nm, 95 nm, 100 nm, 105 nm, 110 nm, 115 nm, 120 nm, 125 nm, 130 nm, 135 nm, 140 nm, 145 nm, 150 nm, 155 nm, 160 nm, 165 nm, 170 nm, 175 nm, 180 nm, 185 nm, 190 nm, 195 nm, 200 nm, 205 nm, 210 nm, 215 nm, 220 nm, or any range having endpoints defined by any two of the foregoing values. For example, in some embodiments, the lipid nanoparticles described herein have an average hydrodynamic diameter of between 50 nm and 200 nm.

[0241] G. Polydispersity In particular, the present disclosure describes compositions, preparations, nanoparticles, and / or nanomaterials having a polydispersity index (PDI) of about 0.01 to about 0.3. In some embodiments, the compositions, preparations, nanoparticles, and / or nanomaterials described herein have a PDI of about 0.01, 0.02, 0.03, 0.04, 0.05, 0.06, 0.07, 0.08, 0.09, 0.1, 0.15, 0.2, 0.25, 0.3, or any range having endpoints defined by any two of the foregoing values. For example, in some embodiments, the compositions, preparations, nanoparticles, and / or nanomaterials described herein have a PDI of about 0.05 to about 0.2, about 0.06 to about 0.1, or about 0.07 to about 0.09.

[0242] In some embodiments, the lipid nanoparticles described herein have a PDI of about 0.01 to about 0.3. In some embodiments, the lipid nanoparticles described herein have a PDI of about 0.01, 0.02, 0.03, 0.04, 0.05, 0.06, 0.07, 0.08, 0.09, 0.1, 0.15, 0.2, 0.25, 0.3, or any range having endpoints defined by any two of the foregoing values. For example, in some embodiments, the lipid nanoparticles described herein have a PDI of about 0.05 to about 0.2, about 0.06 to about 0.1, or about 0.07 to about 0.09.

[0243] H. Encapsulation Efficiency In particular, the present disclosure describes compositions, preparations, nanoparticles, and / or nanomaterials, wherein the encapsulation efficiency of the provided compositions, preparations, nanoparticles, and / or nanomaterials is about 80% to about 100%. In some embodiments, the encapsulation efficiency of the compositions, preparations, nanoparticles, and / or nanomaterials described herein is about 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 95.5%, 96%, 96.5%, 97%, 97.5%, 98%, 98.5%, 99%, 99.5%, 100%, or any range having endpoints defined by any two of the foregoing values. For example, in some embodiments, the encapsulation efficiency of the compositions, preparations, nanoparticles, and / or nanomaterials described herein is about 90% to about 100%, about 95% to about 100%, about 95% to about 98%, or about 95.5% to about 97.5%. In some embodiments, the encapsulation efficiency of the compositions, preparations, nanoparticles, and / or nanomaterials described herein is at least about 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99%.

[0244] In some embodiments, the encapsulation efficiency of the lipid nanoparticles described herein is about 80% to about 100%. In some embodiments, the encapsulation efficiency of the lipid nanoparticles described herein is about 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 95.5%, 96%, 96.5%, 97%, 97.5%, 98%, 98.5%, 99%, 99.5%, 100%, or any range having endpoints defined by any two of the foregoing values. For example, in some embodiments, the encapsulation efficiency of the lipid nanoparticles described herein is about 90% to about 100%, about 95% to about 100%, about 95% to about 98%, or about 95.5% to about 97.5%. In some embodiments, the encapsulation efficiency of the lipid nanoparticles described herein is at least about 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99%.

[0245] I.pKa In particular, the present disclosure describes compositions, preparations, nanoparticles, and / or nanomaterials having a pKa of about 5 to about 9. In some embodiments, the compositions, preparations, nanoparticles, and / or nanomaterials described herein have a pKa of about 5.0, 5.5, 6.0, 6.5, 7.0, 7.5, 8.0, 8.5, or any range having endpoints defined by any two of the foregoing values. In some embodiments, the compositions, preparations, nanoparticles, and / or nanomaterials described herein have a pKa that is about 6.0, 6.1, 6.2, 6.3, 6.4, 6.5, 6.6, 6.7, 6.8, 6.9, 7.0, 7.1, 7.2, 7.3, 7.4, 7.5, 7.6, 7.7, 7.8, 7.9, 8.0, 8.1, 8.2, 8.3, 8.4, 8.5, 8.6, 8.7, 8.8, 8.9, 9.0, or any range having endpoints defined by any two of the foregoing values.

[0246] In some embodiments, the lipid nanoparticles described herein have a pKa of about 5 to about 9. In some embodiments, the lipid nanoparticles described herein have a pKa of about 5.0, 5.5, 6.0, 6.5, 7.0, 7.5, 8.0, 8.5, or any range having endpoints defined by any two of the preceding values. In some embodiments, the lipid nanoparticles described herein have a pKa of about 6.0, 6.1, 6.2, 6.3, 6.4, 6.5, 6.6, 6.7, 6.8, 6.9, 7.0, 7.1, 7.2, 7.3, 7.4, 7.5, 7.6, 7.7, 7.8, 7.9, 8.0, 8.1, 8.2, 8.3, 8.4, 8.5, 8.6, 8.7, 8.8, 8.9, 9.0, or any range having endpoints defined by any two of the preceding values.

[0247] II. Exemplary LNP Preparations The present invention provides compositions, preparations, nanoparticles, and / or nanomaterials comprising lipid nanoparticles. In some embodiments, the lipid nanoparticle preparation comprises about 30 molar percent to about 70 molar percent of an ionizable lipid, about 5 molar percent to about 25 molar percent of a phospholipid, about 25 molar percent to about 45 molar percent of cholesterol, and about 0 molar percent to about 5 molar percent of a conjugate-linker lipid.

[0248] In some embodiments, the lipid nanoparticle preparation comprises about 45 molar percent of ionizable lipid, about 9 molar percent of phospholipid, about 44 molar percent of cholesterol, and about 2 molar percent of conjugate-linker lipid. In some embodiments, the lipid nanoparticle preparation comprises about 50 molar percent of ionizable lipid, about 9 molar percent of phospholipid, about 38 molar percent of cholesterol, and about 3 molar percent of conjugate-linker lipid.

[0249] In some embodiments, the lipid nanoparticle preparation comprises about 47.5 molar percent ionizable lipid, about 10 molar percent phospholipid, about 40 molar percent cholesterol, and about 2.5 molar percent conjugate-linker lipid.

[0250] In some embodiments, the lipid nanoparticle preparation comprises about 40 mole percent to about 60 mole percent of an ionizable lipid of any one of formulas I, II, III, IV-A, IV-B, VA, VB, VI-A, VI-B, VII-A, VII-B, VIII-A, VIII-B, VIII-C, VIII-D, IX-A, IX-B, IX-C, and IX-D, about 5 mole percent to about 15 mole percent of 1-2-distearoyl-sn-glycero-3-phosphocholine, about 1 mole percent to about 5 mole percent of C14PEG2000, and about 30 mole percent to about 47 mole percent of cholesterol, based on the total moles of these four components.

[0251] In some embodiments, the lipid nanoparticle (LNP) preparation comprises a mass ratio of (ionizable lipid, cholesterol, lipid-PEG, and phospholipid):mRNA of about 2:1 to 50:1. In some embodiments, the LNP preparation comprises a mass ratio of about 2:1, about 3:1, about 4:1, about 5:1, about 6:1, about 7:1, about 8:1, about 9:1, about 10:1, about 11:1, about 12:1, about 13:1, about 14:1, about 15:1, about 16:1, about 17:1, about 18:1, about 19:1, about 20:1, about 21:1, about 22:1, about 23:1, about 24:1, about 25:1, about 26:1, about 27:1, about 28:1, about 29:1, about 30:1, about 31:1, about 32:1, about 33:1, about 34:1, about 35:1, about 36:1, about 37:1, about 38:1, about 39:1, about 40:1, about 41:1, about 42:1, about 43:1, about 44:1, about 45:1, about 46:1, about 47:1, about 48:1, about 49:1, about 50:1, about 51:1, about 52:1, about 53:1, about 54:1, about 55:1, about 56:1, about 57:1, about 58:1, about 59:1, about 60:1, about 61:1, about 62:1, about 63:1, about 64:1, about 65:1, about 66:1, 1, about 30:1, about 31:1, about 32:1, about 33:1, about 34:1, about 35:1, about 36:1, about 37:1, about 38:1, about 39:1, about 40:1, about 41:1, about 42:1, about 43:1, about 44:1, about 45:1, about 46:1, about 47:1, about 48:1, about 49:1, about 50:1 mass ratio of (ionizable lipid, cholesterol, lipid-PEG, and phospholipid):mRNA. In some embodiments, the lipid nanoparticle (LNP) preparation comprises a mass ratio of (ionizable lipid, cholesterol, lipid-PEG, and phospholipid):mRNA of about 11.7:1 to 19:1.

[0252] In some embodiments, the lipid nanoparticle preparation comprises a mass ratio of (ionizable lipid, cholesterol, lipid-PEG, and phospholipid):siRNA of about 2:1 to 50:1. In some embodiments, the LNP preparation comprises a mass ratio of about 2:1, about 3:1, about 4:1, about 5:1, about 6:1, about 7:1, about 8:1, about 9:1, about 10:1, about 11:1, about 12:1, about 13:1, about 14:1, about 15:1, about 16:1, about 17:1, about 18:1, about 19:1, about 20:1, about 21:1, about 22:1, about 23:1, about 24:1, about 25:1, about 26:1, about 27:1, about 28:1, about 29:1, about 30:1, about 31:1, about 32:1, about 33:1, about 34:1, about 35:1, about 36:1, about 37:1, about 38:1, about 39:1, about 40:1, about 41:1, about 42:1, about 43:1, about 44:1, about 45:1, about 46:1, about 47:1, about 48:1, about 49:1, about 50:1, about 51:1, about 52:1, about 53:1, about 54:1, about 55:1, about 56:1, about 57:1, about 58:1, about 59:1, about 60:1, about 61:1, about 62:1, about 63:1, about 64:1, about 65:1, about 66:1, about 67: 1, about 30:1, about 31:1, about 32:1, about 33:1, about 34:1, about 35:1, about 36:1, about 37:1, about 38:1, about 39:1, about 40:1, about 41:1, about 42:1, about 43:1, about 44:1, about 45:1, about 46:1, about 47:1, about 48:1, about 49:1, about 50:1 mass ratio of (ionizable lipid, cholesterol, lipid-PEG, and phospholipid):siRNA. In some embodiments, the lipid nanoparticle (LNP) preparation comprises a mass ratio of (ionizable lipid, cholesterol, lipid-PEG, and phospholipid):siRNA of about 11.7:1 to 19:1.

[0253] In some embodiments, the lipid nanoparticle preparation comprises a mass ratio of (ionizable lipid, cholesterol, lipid-PEG, and phospholipid):NA of about 2:1 to 50:1. In some embodiments, the LNP preparation comprises a mass ratio of about 2:1, about 3:1, about 4:1, about 5:1, about 6:1, about 7:1, about 8:1, about 9:1, about 10:1, about 11:1, about 12:1, about 13:1, about 14:1, about 15:1, about 16:1, about 17:1, about 18:1, about 19:1, about 20:1, about 21:1, about 22:1, about 23:1, about 24:1, about 25:1, about 26:1, about 27:1, about 28:1, about 29:1, about 30:1, about 31:1, about 32:1, about 33:1, about 34:1, about 35:1, about 36:1, about 37:1, about 38:1, about 39:1, about 40:1, about 41:1, about 42:1, about 43:1, about 44:1, about 45:1, about 46:1, about 47:1, about 48:1, about 49:1, about 50:1, about 51:1, about 52:1, about 53:1, about 54:1, about 55:1, about 56:1, about 57:1, about 58:1, about 59:1, about 60:1, about 61:1, about 62:1, about 63:1, about 64:1, about 65:1, about 66:1, about 67: 9:1, about 30:1, about 31:1, about 32:1, about 33:1, about 34:1, about 35:1, about 36:1, about 37:1, about 38:1, about 39:1, about 40:1, about 41:1, about 42:1, about 43:1, about 44:1, about 45:1, about 46:1, about 47:1, about 48:1, about 49:1, about 50:1 mass ratio of (ionizable lipid, cholesterol, lipid-PEG, and phospholipid):NA. In some embodiments, the lipid nanoparticle (LNP) preparation comprises a mass ratio of (ionizable lipid, cholesterol, lipid-PEG, and phospholipid):NA of about 11.7:1 to 40:1.

[0254] In some embodiments, the NA comprises a base editor and an sgRNA as described herein. In some embodiments, the mass ratio of the base editor:gRNA is 1:1. In some embodiments, the mass ratio of the base editor:gRNA is 2:1. In some embodiments, the mass ratio of the base editor:gRNA is 3:1. In some embodiments, the mass ratio of the base editor:gRNA is 4:1. In some embodiments, the mass ratio of the base editor:gRNA is 5:1. In some embodiments, the mass ratio of the base editor:gRNA is 6:1. In some embodiments, the mass ratio of the base editor:gRNA is 7:1. In some embodiments, the mass ratio of the base editor:gRNA is 8:1. In some embodiments, the mass ratio of the base editor:gRNA is 9:1. In some embodiments, the mass ratio of the base editor:gRNA is 10:1. In some embodiments, the mass ratio of the base editor:gRNA is 1:2. In some embodiments, the mass ratio of the base editor:gRNA is 1:3. In some embodiments, the mass ratio of base editor:gRNA is 1:4. In some embodiments, the mass ratio of base editor:gRNA is 1:5. In some embodiments, the mass ratio of base editor:gRNA is 1:6. In some embodiments, the mass ratio of base editor:gRNA is 1:7. In some embodiments, the mass ratio of base editor:gRNA is 1:8. In some embodiments, the mass ratio of base editor:gRNA is 1:9. In some embodiments, the mass ratio of base editor:gRNA is 1:10.

[0255] III. Pharmaceutical Compositions The present invention provides compositions, preparations, nanoparticles, and / or nanomaterials, including pharmaceutical compositions. In particular, in some embodiments, the pharmaceutical compositions include the lipid nanoparticles and lipid nanoparticle preparations described herein. For example, in some embodiments, the lipid nanoparticles and lipid nanoparticle preparations described herein may be formulated in whole or in part as pharmaceutical compositions.

[0256] In some embodiments, a pharmaceutical composition may comprise one or more nanoparticle compositions described herein. For example, a pharmaceutical composition may comprise one or more nanoparticle compositions comprising one or more different therapeutic and / or prophylactic agents, including, but not limited to, different types of one or more nucleic acids, or may encode different agents. In some embodiments, a pharmaceutical composition comprises one or more pharma- ceutically acceptable excipients or auxiliary ingredients, including, but not limited to, a pharma- ceutically acceptable carrier.

[0257] The pharmaceutical composition can be administered to a subject. In some embodiments, the pharmaceutical composition is administered as described herein. In some in vivo approaches, the nanoparticle composition disclosed herein is administered to a subject in a therapeutically effective amount as described herein.

[0258] In some embodiments, one skilled in the art will be able to devise appropriate dosage levels and dosing regimens using the pharmaceutical compositions described herein for the treatment of various conditions in various patients, taking into account the therapeutic background, age, and general health of the recipient. For example, in some embodiments, the selected dosage will depend on the desired therapeutic effect, the route of administration, and the desired duration of treatment. In some embodiments, dosage levels of about 0.001 mg to about 5 mg of nucleic acid per kg of body weight are typically administered to a mammal for each dose. More specifically, in some embodiments, the preferred dosage of nucleic acid in the disclosed nanoparticles is about 0.1 mg / kg to about 1.0 mg / kg. In the disclosed nanoparticles, dosage levels of about 0.2 mg to about 100 mg of the four components (ionizable lipid, cholesterol, conjugate-linker conjugate, and phospholipid) per kg of body weight are typically administered to a mammal. More specifically, in some embodiments, the preferred dosage of the disclosed nanoparticles is about 0.5 mg / kg to about 5 mg / kg of the four components per kg of body weight.

[0259] In some embodiments, the pharmaceutical compositions described herein are administered locally, for example, by direct injection into the site to be treated. Typically, the injection causes a local concentration increase of the composition that is higher than can be achieved by systemic administration. In some embodiments, the pharmaceutical compositions described herein can be combined with the matrices described herein to help create a local concentration increase of the polypeptide composition by reducing passive diffusion of the polypeptide from the site to be treated.

[0260] A. Preparations for Parenteral Administration In some embodiments, compositions, preparations, nanoparticles, and / or nanomaterials disclosed herein, including those containing lipid nanoparticles, are administered by parenteral injection in an aqueous solution. In some embodiments, the preparations may also be in the form of a suspension or emulsion. In general, a pharmaceutical composition is provided that contains an effective amount of lipid nanoparticles, and optionally includes a pharma- ceutically acceptable diluent, preservative, solubilizer, emulsifier, adjuvant, and / or carrier. Such compositions optionally include one or more of diluents, sterile water, buffered saline of various buffer contents (e.g., Tris-HCl, acetate, phosphate), pH, and ionic strength; and additives such as detergents and solubilizers (e.g., TWEEN® 20 (polysorbate-20), TWEEN 80 (polysorbate-80)), antioxidants (e.g., ascorbic acid, sodium metabisulfite), and preservatives (e.g., Thimersol, benzyl alcohol), and bulking substances (e.g., lactose, mannitol). Examples of non-aqueous solvents or vehicles are propylene glycol, polyethylene glycol, vegetable oils such as olive oil and corn oil, gelatin, and injectable organic esters such as ethyl oleate.The formulation can be lyophilized and redissolved / resuspended immediately before use.The formulation can be sterilized, for example, by filtration through a bacteria-retaining filter, by incorporating a sterilizing agent into the composition, by irradiating the composition, or by heating the composition.

[0261] B. Controlled Delivery Polymer Matrices In some embodiments, the compositions, preparations, nanoparticles, and / or nanomaterials disclosed herein may also be administered in controlled release formulations. In some embodiments, controlled release polymeric devices may be fabricated for extended release systemically after implantation of the polymeric device (rods, cylinders, films, disks, etc.) or injection (microparticles, etc.). In some embodiments, the matrix may be in the form of microparticles, such as microspheres. In some embodiments, the drug is dispersed within a solid polymer matrix or microcapsules. In some embodiments, the core is of a different material than the polymeric shell of any of the described compositions, preparations, nanoparticles, and / or nanomaterials. In some embodiments, the peptide is dispersed or suspended in the core, which may be liquid or solid in nature, of any of the described compositions, preparations, nanoparticles, and / or nanomaterials. Unless otherwise specifically defined herein, microparticles, microspheres, and microcapsules are used interchangeably. In some embodiments, the polymer may be cast as a thin slab or film ranging from nanometers to four centimeters, a powder produced by grinding or other standard techniques, or even a gel, such as a hydrogel.

[0262] In some embodiments, non-biodegradable matrices are used to deliver the described compositions, preparations, nanoparticles, and / or nanomaterials. In some embodiments, biodegradable matrices are used to deliver the described compositions, preparations, nanoparticles, and / or nanomaterials. In some embodiments, biodegradable matrices are preferred. In some embodiments, the biodegradable matrices include natural or synthetic polymers. In some embodiments, synthetic polymers are preferred due to better characterization of degradation and release profiles. In some embodiments, the polymer is selected based on the period over which release is desired. In some embodiments, linear release may be most useful, while in other embodiments, pulsed or "bulk release" may provide more effective results. In some embodiments, the polymer may be in the form of a hydrogel (typically absorbing up to about 90% water by weight), which may be optionally crosslinked with multivalent ions or polymers.

[0263] The matrix can be formed by solvent evaporation, spray drying, solvent extraction, and other methods known to those skilled in the art. Bioerodible microspheres can be prepared using any of the methods developed to make microspheres for drug delivery, for example, as described by Mathiowitz and Langer, J. Controlled Release, 5:13-22 (1987); Mathiowitz, et al., Reactive Polymers, 6:275-283 (1987); and Mathiowitz, et al., J. Appl. Polymer Sci., 35:755-774 (1988), the entire disclosures of which are incorporated herein by reference.

[0264] In some embodiments, the described compositions, preparations, nanoparticles, and / or nanomaterials can be formulated for local release to treat the area of ​​implantation or injection, and will typically deliver a much smaller dose than for whole body treatment or systemic delivery. They may be implanted or injected subcutaneously, into muscle, fat, or swallowed.

[0265] C. Cargo In particular, the present invention provides compositions, preparations, nanoparticles, and / or nanomaterials comprising the cargo described herein. In some embodiments, the compositions, preparations, nanoparticles, and / or nanomaterials comprise a therapeutic or prophylactic agent for delivery to a subject. In some embodiments, the therapeutic or prophylactic agent is encapsulated by the lipid nanoparticle. In some embodiments, the lipid nanoparticles are loaded with one or more nucleic acids.

[0266] D. Therapeutic and / or Prophylactic Agents The cargo delivered via the LNP composition can be a biologically active agent. In some embodiments, the cargo can be an mRNA, a guide RNA (gRNA), a nucleic acid, an RNA-guided DNA binder, an expression vector, a template nucleic acid, an antibody (e.g., monoclonal, chimeric, humanized, nanobody, and fragments thereof), cholesterol, a hormone, a peptide, a protein, a chemotherapeutic agent and other types of antineoplastic agents, a low molecular weight drug, a vitamin, a cofactor, a nucleoside, a nucleotide, an oligonucleotide, an enzymatic nucleic acid, an antisense nucleic acid, a triplex forming oligonucleotide, an antisense DNA or RNA composition, a chimeric DNA:RNA composition, an allozyme, an aptamer, a ribozyme, a decoy and an analog thereof, a plasmid and other types of vectors. The biologically active agent may be or include one or more biologically active agents, such as small nucleic acid molecules, RNAi agents, short interfering nucleic acids (siNA), short interfering RNAs (siRNA), double-stranded RNAs (dsRNA), microRNAs (miRNA), short hairpin RNAs (shRNA), "self-replicating RNA" molecules (capable of encoding replicase enzyme activity and inducing its own replication or amplification in vivo), peptide nucleic acids (PNAs), locked nucleic acid ribonucleotides (LNAs), morpholino nucleotides, threose nucleic acids (TNAs), glycol nucleic acids (GNAs), sisiRNAs (small internal segmented interfering RNAs), and iRNAs (asymmetric interfering RNAs). The above list of biologically active agents is illustrative only and is not intended to be limiting. Such compounds may be purified or partially purified, naturally occurring or synthetic, and chemically modified.

[0267] The cargo delivered via the LNP composition may be an RNA, such as an mRNA molecule, encoding a protein of interest. For example, in some embodiments, an mRNA for expressing a protein, such as green fluorescent protein (GFP), an RNA-guided DNA binder, or a Cas nuclease, is described herein. LNP compositions are provided that include a Cas nuclease mRNA, e.g., a class 2 Cas nuclease mRNA, that allows expression of a class 2 Cas nuclease, such as Cas9 or a Cpfl protein, in a cell. Additionally, the cargo may contain one or more guide RNAs or a nucleic acid encoding the guide RNA. For example, a template nucleic acid for repair or recombination may also be included in the composition, or the template nucleic acid may be used in the methods described herein. In some embodiments, the cargo optionally includes an mRNA encoding Streptococcus pyogenes Cas9 and an S. pyogenes gRNA. In some embodiments, the cargo optionally includes an mRNA encoding Neisseria meningitidis Cas9 and an nme gRNA.

[0268] "mRNA" refers to a polynucleotide and contains an open reading frame that can be translated into a polypeptide (i.e., can serve as a substrate for translation by ribosomes and amino-acylated tRNAs). An mRNA can contain a phosphate-sugar backbone that includes ribose residues or analogs thereof, e.g., 2'-methoxyribose residues. In some embodiments, the sugars of the mRNA phosphate sugar backbone consist essentially of ribose residues, 2'-methoxyribose residues, or combinations thereof. Generally, an mRNA does not contain a substantial amount of thymidine residues (e.g., 0 residues or less than 30, 20, 10, 5, 4, 3, or 2 thymidine residues; or a thymidine content of less than 10%, 9%, 8%, 7%, 6%, 5%, 4%, 4%, 3%, 2%, 1%, 0.5%, 0.2%, or 0.1%). An mRNA can contain modified uridines at some or all of its uridine positions.

[0269] E. CRISPR / Cas cargo In some embodiments, the disclosed compositions, preparations, nanoparticles, and / or nanomaterials comprise an mRNA encoding an RNA-guided DNA binder, such as a Cas nuclease. In certain embodiments, the disclosed compositions, preparations, nanoparticles, and / or nanomaterials comprise an mRNA encoding a class 2 Cas nuclease, such as S. pyogenes Cas9.

[0270] As used herein, "RNA-guided DNA binding agent" refers to a polypeptide or a complex of polypeptides having RNA and DNA binding activity, or a DNA-binding subunit of such a complex, where the DNA-binding activity is sequence-specific and dependent on the sequence of the RNA. Exemplary RNA-guided DNA binding agents include Cas cleavase / nickases and their inactivated forms (dCas DNA binding agents). "Cas nuclease" as used herein encompasses Cas cleavase, Cas nickase, and dCas DNA binding agents. Cas cleavase / nickases and dCas DNA binding agents include the Csm or Cmr complex of type III CRISPR systems, its CaslO, Csml, or Cmr2 subunits, the Cascade complex of type I CRISPR systems, its Cas3 subunit, and class 2 Cas nuclease. As used herein, "class 2 Cas nuclease" is a single-chain polypeptide having RNA-guided DNA binding activity. Class 2 Cas nucleases include Class 2 Cas cleavage enzymes / nickases that further have RNA-guided DNA cleavage enzyme or nickase activity (e.g., H840A, D10A, or N863A variants), and Class 2 dCas DNA binders in which the cleavage enzyme / nickase activity has been inactivated. Class 2 Cas nucleases include, for example, Cas9, Cpfl, C2cl, C2c2, C2c3, HF Cas9 (e.g., N497A, R661A, Q695A, Q926A variants), HypaCas9 (e.g., N692A, M694A, Q695A, H698A variants), eSPCas9(1.0) (e.g., K810A, K1003A, R1060A variants), and eSPCas9(1.1) (e.g., K848A, K1003A, R1060A variants) proteins and modifications thereof. The Cpfl protein, Zetsche et al., Cell, 163:1-13 (2015), is homologous to Cas9 and contains a RuvC-like nuclease domain. The Cpfl sequences of Zetsche are incorporated herein by reference in their entirety, see, e.g., Tables S1 and S3 of Zetsche.See, e.g., Makarova et al., Nat Rev Microbiol, 13(11):722-36 (2015); Shmakov et al., Molecular Cell, 60:385-397 (2015), the contents of which are incorporated herein in their entireties.

[0271] As used herein, "ribonucleoprotein" (RNP) or "RNP complex" refers to a guide RNA comprising an RNA-guided DNA-binding agent, such as a Cas nuclease, such as a Cas cleavase, a Cas nickase, or a dCas DNA-binding agent (e.g., Cas9). In some embodiments, the guide RNA guides an RNA-guided DNA-binding agent, such as Cas9, to a target sequence, the guide RNA hybridizes to the target sequence, and an agent binds to the target sequence, where the agent is a cleavase or nickase and can perform cleavage or nicking after binding.

[0272] In some embodiments, the cargo of the LNP composition comprises at least one guide RNA comprising a guide sequence that guides an RNA-guided DNA-binding agent, which may be a nuclease (e.g., a Cas nuclease, such as Cas9), to a target DNA. The gRNA may guide the Cas nuclease or a class 2 Cas nuclease to a target sequence on a target nucleic acid molecule. In some embodiments, the gRNA binds to a class 2 Cas nuclease, thereby providing specificity for cleavage. In some embodiments, the gRNA and the Cas nuclease may form a ribonucleoprotein (RNP), e.g., a CRISPR / Cas complex, such as a CRISPR / Cas9 complex. In some embodiments, the CRISPR / Cas complex may be a type II CRISPR / Cas9 complex. In some embodiments, the CRISPR / Cas complex may be a type V CRISPR / Cas complex, such as a Cpfl / guide RNA complex. The Cas nuclease and the cognate gRNA may pair. The gRNA scaffold structure that pairs with each Class 2 Cas nuclease varies depending on the particular CRISPR / Cas system.

[0273] "Guide RNA", "gRNA", and simply "guide" are used interchangeably herein to refer to either crRNA (also known as CRISPR RNA) or a combination of crRNA and trRNA (also known as tracrRNA). Guide RNA may include modified RNA as described herein. crRNA and trRNA may be associated as a single-stranded RNA molecule (single guide RNA, sgRNA) or in two separate RNA molecules (dual guide RNA, dgRNA). "Guide RNA" or "gRNA" refers to each type. trRNA may be a naturally occurring sequence or a trRNA sequence with modifications or mutations compared to the naturally occurring sequence.

[0274] As used herein, "guide sequence" refers to a sequence in a guide RNA that is complementary to a target sequence and functions to direct the guide RNA to the target sequence for binding or modification (e.g., cleavage) by an RNA-guided DNA binding agent. A "guide sequence" may also be referred to as a "targeting sequence" or a "spacer sequence." A guide sequence may be, for example, 20 base pairs in length for Streptococcus pyogenes (i.e., Spy Cas9) and related Cas9 homologs / orthologues. Shorter or longer sequences, such as, for example, 15, 16, 17, 18, 19, 21, 22, 23, 24, or 25 nucleotides in length, can also be used as a guide. In some embodiments, the target sequence is, for example, within a gene or on a chromosome, and is complementary to the guide sequence. In some embodiments, the degree of complementarity or identity between a guide sequence and its corresponding target sequence may be about or at least 75%, 80%, 85%, 90%, 95%, 96%, 97%, 98%, 99%, or 100%. In some embodiments, the guide sequence and the target region may be 100% complementary or identical over a region of at least 15, 16, 17, 18, 19, or 20 contiguous nucleotides. In other embodiments, the guide sequence and the target region may contain at least one mismatch. For example, the guide sequence and the target sequence may contain 1, 2, 3, or 4 mismatches, and the total length of the target sequence is at least 17, 18, 19, 20, or more base pairs. In some embodiments, the guide sequence and the target region may contain 1-4 mismatches, and the guide sequence comprises at least 17, 18, 19, 20, or more nucleotides. In some embodiments, the guide sequence and the target region may contain 1, 2, 3, or 4 mismatches, and the guide sequence comprises 20 nucleotides.

[0275] The target sequence of an RNA-guided DNA-binding protein, such as a Cas protein, includes both the positive and negative strands of genomic DNA (i.e., the given sequence and the reverse complement of the sequence), since the nucleic acid substrate of the Cas protein is a double-stranded nucleic acid. Thus, when a guide sequence is said to be "complementary to a target sequence," it should be understood that the guide RNA can be directed to bind to the reverse complement of the target sequence. Thus, in some embodiments, when the guide sequence binds to the reverse complement of the target sequence, the guide sequence is identical to a particular nucleotide of the target sequence (e.g., the target sequence without the PAM), except for the substitution of U for T in the guide sequence.

[0276] The length of the targeting sequence depends on the CRISPR / Cas system and components used. For example, various class 2 Cas nucleases from various bacterial species have different optimal targeting sequence lengths. Thus, the targeting sequence may comprise 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 35, 40, 45, 50, or more than 50 nucleotides in length. In some embodiments, the targeting sequence length is 0, 1, 2, 3, 4, or 5 nucleotides longer or shorter than the guide sequence of the naturally occurring nucleotide sequence.

[0277] CRISPR / Cas systems In certain embodiments, the Cas nuclease and the gRNA scaffold will be derived from the same CRISPR / Cas system. In some embodiments, the targeting sequence may comprise or consist of 18-24 nucleotides. In some embodiments, the targeting sequence may comprise or consist of 19-21 nucleotides. In some embodiments, the targeting sequence may comprise or consist of 20 nucleotides.

[0278] In some embodiments, the sgRNA is a "Cas9 sgRNA" capable of mediating RNA-guided DNA cleavage by a Cas9 protein. In some embodiments, the sgRNA is a "Cpfl sgRNA" capable of mediating RNA-guided DNA cleavage by a Cpfl protein. In some embodiments, the gRNA comprises sufficient crRNA and tracrRNA to form an active complex with a Cas9 protein and mediate RNA-guided DNA cleavage. In some embodiments, the gRNA comprises sufficient crRNA to form an active complex with a Cpfl protein and mediate RNA-guided DNA cleavage. See Zetsche 2015.

[0279] Certain embodiments of the invention also provide nucleic acids, e.g., expression cassettes, encoding the gRNAs described herein. "Guide RNA nucleic acid" is used herein to refer to guide RNAs (e.g., sgRNAs or dgRNAs) and guide RNA expression cassettes, which are nucleic acids that encode one or more guide RNAs.

[0280] Certain embodiments of the present disclosure also provide for delivery of adenine base editors ("ABEs") using the LNP compositions, preparations, nanoparticles, and / or nanomaterials described herein. ABEs and methods of using same are described, for example, in U.S. Patent No. 10,113,163 and U.S. Patent Publication No. 2021 / 0130805, the contents of each of which are incorporated herein by reference in their entirety.

[0281] Certain embodiments of the present disclosure also provide for delivery of cytosine base editors ("CBEs") using the LNP compositions, preparations, nanoparticles, and / or nanomaterials described herein. CBEs and methods of using same are described, for example, in U.S. Patent Nos. 10,167,457 and 9,840,699, the contents of each of which are incorporated herein by reference in their entirety.

[0282] The term "base editor (BE)" or "nucleobase editor (NBE)" refers to an agent comprising a polypeptide capable of making modifications to bases (e.g., A, T, C, G, or U) in a nucleic acid sequence (e.g., DNA or RNA). In some embodiments, the base editor is capable of deaminating a base in a nucleic acid. In some embodiments, the base editor is capable of deaminating a base in a DNA molecule. In some embodiments, the base editor is capable of deaminating an adenine (A) in DNA. In some embodiments, the deaminase is a cytosine deaminase or a cytidine deaminase. In some embodiments, the base editor is a fusion protein comprising a nucleic acid programmable DNA binding protein (napDNAbp) fused to an adenosine deaminase. In some embodiments, the base editor is a Cas9 protein fused to an adenosine deaminase. In some embodiments, the base editor is a Cas9 nickase (nCas9) fused to an adenosine deaminase. In some embodiments, the base editor is a nuclease-inactive Cas9 (dCas9) fused to an adenosine deaminase. In some embodiments, the base editor is fused to an inhibitor of base excision repair (e.g., a UGI domain, or a dISN domain). In some embodiments, the fusion protein comprises a Cas9 nickase fused to a deaminase and an inhibitor of base excision repair (e.g., a UGI domain or a dISN domain). The term "nucleic acid-programmed DNA binding protein" or "napDNAbp" refers to a protein that associates with a nucleic acid (e.g., DNA or RNA), such as a guide nucleic acid that directs the napDNAbp to a specific nucleic acid sequence. For example, a Cas9 protein can associate with a guide RNA that directs the Cas9 protein to a specific DNA sequence that has complementarity to the guide RNA. In some embodiments, the napDNAbp is a class 2 microbial CRISPR-Cas effector. In some embodiments, the napDNAbp is a Cas9 domain, e.g., a nuclease-active Cas9, Cas9 nickase (nCas9), or a nuclease-inactive Cas9 (dCas9).Examples of nucleic acid programmed DNA binding proteins include, but are not limited to, Cas9 (e.g., dCas9 and nCas9), CasX, CasY, Cpf1, C2c1, C2c2, C2C3, and Argonaute. However, it should be understood that nucleic acid programmed DNA binding proteins can also include nucleic acid programmed proteins that bind to RNA. For example, napDNAbp can associate with a nucleic acid that guides napDNAbp to RNA. Other nucleic acid programmed DNA binding proteins are also within the scope of this disclosure, although they may not be specifically listed in this disclosure.

[0283] F. Modified RNA In certain embodiments, the disclosed compositions, preparations, nanoparticles, and / or nanomaterials comprise modified nucleic acids, including modified RNA.

[0284] Modified nucleosides or nucleotides can be present in an RNA, such as a gRNA or an mRNA. For example, a gRNA or an mRNA that includes one or more modified nucleosides or nucleotides is referred to as a "modified" RNA to account for the presence of one or more non-natural and / or naturally occurring components or arrangements that are used in place of or in addition to the standard A, G, C, and U residues. In some embodiments, a modified RNA is synthesized with non-standard nucleosides or nucleotides and is referred to herein as "modified."

[0285] Modified nucleosides and nucleotides can include one or more of: (i) alteration, e.g., replacement, of one or both of one or more of the non-linked and / or linked phosphate oxygens in a phosphodiester backbone linkage (exemplary backbone modifications); (ii) alteration, e.g., replacement, of a component of the ribose sugar, e.g., the 2' hydroxyl on the ribose sugar (exemplary sugar modifications); (iii) wholesale replacement of a phosphate moiety with a "dephospho" linker (exemplary backbone modifications); (iv) modification or replacement of a naturally occurring nucleobase, including a non-canonical nucleobase (exemplary base modifications); (v) replacement or modification of the ribose-phosphate backbone (exemplary backbone modifications); (vi) modification of the 3' or 5' end of an oligonucleotide, e.g., removal, modification, or replacement of a terminal phosphate group or conjugation of a moiety, cap, or linker (such 3' or 5' cap modifications can include sugar and / or backbone modifications); and (vii) modification or replacement of a sugar (exemplary sugar modifications). Certain embodiments include a 5'-end modification to the mRNA, gRNA, or nucleic acid. Certain embodiments include a 3'-end modification to the mRNA, gRNA, or nucleic acid. The modified RNA may contain a 5'-end and a 3'-end modification. The modified RNA may contain one or more modified residues at a non-terminal position. In certain embodiments, the gRNA includes at least one modified residue. In certain embodiments, the mRNA includes at least one modified residue.

[0286] Unmodified nucleic acids may be susceptible to degradation, for example, by intracellular nucleases or those found in serum. For example, nucleases may hydrolyze phosphodiester bonds of nucleic acids. Thus, in one aspect, the RNAs (e.g., mRNA, gRNA) described herein may contain one or more modified nucleosides or nucleotides, for example, to introduce stability against intracellular or serum-based nucleases. In some embodiments, the modified gRNA molecules described herein may exhibit a reduced innate immune response when introduced into a cell population, both in vivo and ex vivo. The term "innate immune response" includes cellular responses to exogenous nucleic acids, including single-stranded nucleic acids, including induction of cytokine (particularly interferon) expression and release and cell death.

[0287] Thus, in some embodiments, the RNA or nucleic acid in the compositions, preparations, nanoparticles, and / or nanomaterials of the present disclosure comprises at least one modification that confers increased or enhanced stability to the nucleic acid, including, for example, improved resistance to nuclease digestion in vivo. As used herein, the terms "modification" and "modified", when such terms relate to the nucleic acids provided herein, preferably include at least one alteration that enhances stability and makes the RNA or nucleic acid more stable (e.g., resistant to nuclease digestion) than a wild-type or naturally occurring version of the RNA or nucleic acid. As used herein, the terms "stable" and "stability", when such terms relate to the nucleic acids of the present invention, particularly with respect to RNA, refer to, for example, increased or enhanced resistance to degradation by nucleases (i.e., endonucleases or exonucleases) that would normally degrade such RNA. Increased stability can include, for example, reduced susceptibility to hydrolysis or other destruction by endogenous enzymes (e.g., endonucleases or exonucleases) or conditions in the target cell or tissue, thereby increasing or enhancing the presence of such RNA in the target cell, tissue, subject, and / or cytoplasm. The stabilized RNA molecules provided herein exhibit longer half-lives compared to their naturally occurring unmodified counterparts (e.g., wild-type versions of mRNA). Also contemplated by the terms "modification" and "modified," as such terms relate to the mRNA of the LNP compositions disclosed herein, are modifications that improve or enhance translation of the mRNA nucleic acid, including, for example, the inclusion of sequences that function in the initiation of protein translation (e.g., Kozac consensus sequences). (Kozak, M., Nucleic Acids Res 15(20):8125-48 (1987), the entire contents of which are incorporated herein by reference).

[0288] In some embodiments, the RNA or nucleic acid of the disclosed compositions, preparations, nanoparticles, and / or nanomaterials disclosed herein is chemically or biologically modified to make it more stable. Exemplary modifications to RNA include base depletion (e.g., by deletion or by replacing one nucleotide with another), or base modification, e.g., chemical modification of the base. The phrase "chemical modification" as used herein includes modifications that introduce chemical properties different from those found in naturally occurring RNA, e.g., covalent modifications such as the introduction of modified nucleotides (e.g., including nucleotide analogs, or pendant groups not naturally found in such RNA molecules).

[0289] In some embodiments of backbone modification, the phosphate group of the modified residue may be modified by replacing one or more oxygens with different substituents. Furthermore, modified residues, such as those present in modified nucleic acids, may include extensive replacement of unmodified phosphate moieties with modified phosphate groups as described herein. In some embodiments, backbone modification of the phosphate backbone may include modifications that result in either uncharged linkers or charged linkers with asymmetric charge distribution. Examples of modified phosphate groups include phosphorothioates, phosphoroselenates, boranophosphates, boranophosphate esters, hydrogen phosphonates, phosphoramidates, alkyl or aryl phosphonates, and phosphotriesters. The phosphate atom in an unmodified phosphate group is achiral. However, replacement of one of the non-bridging oxygens with one of the atoms or groups of atoms described above can make the phosphorus atom chiral. The stereogenic phosphorus atom may have either the "R" configuration (herein Rp) or the "S" configuration (herein Sp). The backbone can also be modified by replacement of bridging oxygens (i.e., oxygens linking the phosphate to the nucleoside) with nitrogen (bridging phosphoramidates), sulfur (bridging phosphorothioates), and carbon (bridging methylene phosphonates). Replacement can occur at either bridging oxygen or at both bridging oxygens. The phosphate group can be replaced by a non-phosphorus-containing linking group in certain backbone modifications. In some embodiments, the charged phosphate group can be replaced by a neutral moiety. Examples of moieties that can replace the phosphate group can include, but are not limited to, for example, methylphosphonate, hydroxylamino, siloxane, carbonate, carboxymethyl, carbamate, amide, thioether, ethylene oxide linker, sulfonate, sulfonamide, thioformacetal, formacetal, oxime, methyleneimino, methylenemethylimino, methylenehydrazo, methylenedimethylhydrazo, methyleneoxymethylimino.

[0290] G.mRNA In some embodiments, the compositions, preparations, nanoparticles, and / or nanomaterials of the present disclosure include an mRNA that includes an open reading frame (ORF) encoding an RNA-guided DNA-binding agent, such as a Cas nuclease or class 2 Cas nuclease, as described herein. In some embodiments, an mRNA is provided, used, or administered that includes an ORF encoding an RNA-guided DNA-binding agent, such as a Cas nuclease or class 2 Cas nuclease. The mRNA may include one or more of a 5' cap, a 5' untranslated region (UTR), a 3' UTR, and a polyadenine tail. The mRNA may include an open reading frame that has been modified, for example, to encode a nuclear localization sequence or to use alternative codons to encode a protein.

[0291] The mRNA in the disclosed compositions, preparations, nanoparticles, and / or nanomaterials may, for example, encode a normally secreted hormone, an enzyme, a receptor, a polypeptide, a peptide, or other protein of interest. In one embodiment of the invention, the mRNA may optionally have chemical or biological modifications that, for example, improve the stability and / or half-life of such mRNA or improve or enhance protein production.

[0292] In addition, suitable modifications include alterations in one or more nucleotides of a codon such that the codon codes for the same amino acid but is more stable than the codon found in the wild-type version of the mRNA. For example, an inverse relationship between RNA stability and a higher number of cytidine (C) and / or uridine (U) residues has been shown, and RNA lacking C and U residues has been found to be stable against most RNases (Heidenreich, et al. J Biol Chem 269, 2131-8 (1994), the entire disclosure of which is incorporated herein by reference). In some embodiments, the number of C and / or U residues in the mRNA sequence is reduced. In another embodiment, the number of C and / or U residues is reduced by replacing one codon that codes for a particular amino acid with another codon that codes for the same or related amino acid. Contemplated modifications to the mRNA nucleic acid of the present invention also include the incorporation of pseudouridine. The incorporation of pseudouridine into the mRNA nucleic acid of the present invention may enhance stability and translational capacity, as well as reduce immunogenicity in vivo. See, for example, Kariko, K., et al., Molecular Therapy 16(11):1833-1840 (2008), the entire contents of which are incorporated herein by reference. Substitutions and modifications to the mRNA of the present invention can be made by methods readily known to those of skill in the art.

[0293] Constraints on reducing the number of C and U residues in a sequence will likely be greater in the coding regions of an mRNA compared to untranslated regions (i.e., it will likely not be possible to eliminate all of the C and U residues present in a message while still retaining the capacity of the message to code for a desired amino acid sequence). However, the degeneracy of the genetic code presents opportunities to reduce the number of C and / or U residues present in a sequence while maintaining the same coding capacity (i.e., depending on which amino acids are coded for by the codons, several different possibilities for modification of an RNA sequence may be possible).

[0294] The term modification also includes, for example, the incorporation of non-nucleotide linkages or modified nucleotides into the mRNA sequences of the invention (e.g., modifications to one or both of the 3' and 5' ends of an mRNA molecule encoding a functional secreted protein or enzyme). Such modifications include the addition of bases to the mRNA sequence (e.g., inclusion of a polyA tail or a longer polyA tail), alteration of the 3' UTR or 5' UTR, complexing the mRNA with an agent (e.g., a protein or a complementary nucleic acid molecule), and the inclusion of elements that alter the structure of the mRNA molecule (e.g., forming a secondary structure).

[0295] PolyA tails are believed to stabilize natural messengers. Thus, in one embodiment, long polyA tails can be added to mRNA molecules, thus making the mRNA more stable. PolyA tails can be added using a variety of techniques recognized in the art. For example, long polyA tails can be added to synthetic or in vitro transcribed mRNAs using polyA polymerase (Yokoe, et al. Nature Biotechnology. 1996;14:1252-1256, the entire contents of which are incorporated herein by reference). Transcription vectors can also encode long polyA tails. In addition, polyA tails can be added by direct transcription from PCR products. In one embodiment, the length of the polyA tail is at least about 90, 200, 300, 400, at least 500 nucleotides. In one embodiment, the length of the polyA tail is adjusted to control the stability of the modified mRNA molecules of the present invention, and thus the transcription of the protein. For example, the length of the polyA tail can affect the half-life of an mRNA molecule, and so the length of the polyA tail can be adjusted to alter the level of resistance of the mRNA to nucleases, thereby controlling the time course of protein expression in a cell. In one embodiment, the stabilized mRNA molecule is sufficiently resistant to in vivo degradation (e.g., by nucleases) that it can be delivered to a target cell without the use of a transfer vehicle.

[0296] In some embodiments, the mRNA can be modified by incorporation of 3' and / or 5' untranslated (UTR) sequences not naturally found in the wild-type mRNA. In one embodiment, to modify, 3' and / or 5' flanking sequences naturally adjacent to the mRNA and encoding a second unrelated protein can be incorporated into the nucleotide sequence of the mRNA molecule encoding a therapeutic or functional protein. For example, to increase the stability of the sense mRNA molecule, 3' or 5' sequences from a stable mRNA molecule (e.g., globin, actin, GAPDH, tubulin, histones, or citric acid cycle enzymes) can be incorporated into the 3' and / or 5' region of the sense mRNA nucleic acid molecule. See, for example, US2003 / 0083272, the entire contents of which are incorporated herein by reference. A more detailed description of mRNA modifications can be found in US2017 / 0210698A1, pages 57-68, the entire contents of which are incorporated herein by reference.

[0297] H. Template Nucleic Acid The compositions, preparations, nanoparticles, and / or nanomaterials and methods disclosed herein may include a template nucleic acid. The template may be used to modify or insert a nucleic acid sequence at or near a target site of an RNA-guided DNA-binding protein, such as a Cas nuclease, e.g., class 2 Cas nuclease. In some embodiments, the method includes introducing the template into a cell. In some embodiments, a single template may be provided. In some embodiments, two or more templates may be provided so that editing can be performed at two or more target sites. For example, different templates may be provided to edit a single gene in a cell, or two different genes in a cell.

[0298] In some embodiments, the template may be used in homologous recombination. In some embodiments, the template sequence or a portion of the template sequence may be incorporated into the target nucleic acid molecule by homologous recombination. In some embodiments, the template may be used in homology-directed repair, which involves the invasion of a DNA strand at the nucleic acid cleavage site. In some embodiments, the homology-directed repair may result in the inclusion of the template sequence in the edited target nucleic acid molecule. In some embodiments, the template may be used in gene editing mediated by non-homologous end joining. In some embodiments, the template sequence does not have similarity to the nucleic acid sequence near the cleavage site. In some embodiments, the template or a portion of the template sequence is incorporated. In some embodiments, the template includes adjacent inverted terminal repeat (ITR) sequences.

[0299] In some embodiments, the template sequence may correspond to, comprise, or consist of an endogenous sequence of the target cell. The template sequence may also, or alternatively, correspond to, comprise, or consist of an exogenous sequence of the target cell. As used herein, the term "endogenous sequence" refers to a sequence that is native to the cell. The term "exogenous sequence" refers to a sequence that is not native to the cell or that is at a different location than its native location in the genome of the cell. In some embodiments, the endogenous sequence may be a genomic sequence of the cell.

[0300] In some embodiments, the endogenous sequence may be a chromosomal or extrachromosomal sequence, hi some embodiments, the endogenous sequence may be a plasmid sequence of the cell.

[0301] In some embodiments, the template contains ssDNA or dsDNA containing flanking inverted terminal repeat (ITR) sequences. In some embodiments, the template is provided as a vector, a plasmid, a minicircle, a nanocircle, or a PCR product.

[0302] In some embodiments, the nucleic acid is purified. In some embodiments, the nucleic acid is purified using a precipitation method (e.g., LiCl precipitation, alcohol precipitation, or equivalent methods, e.g., as described herein). In some embodiments, the nucleic acid is purified using a chromatography-based method, such as an HPLC-based method or equivalent methods (e.g., as described herein). In some embodiments, the nucleic acid is purified using both a precipitation method (e.g., LiCl precipitation) and an HPLC-based method. In some embodiments, the nucleic acid is purified by tangential flow filtration (TFF).

[0303] IV. Methods of Producing LNPs Methods for producing lipid nanoparticles are known in the art. In some embodiments, the compositions, preparations, nanoparticles, and / or nanomaterials described are produced using microfluidics. For example, exemplary methods for forming lipid nanoparticles using microfluidics are described by Leung, AKK, et al., J Phys Chem, 116: 18440-18450 (2012), Chen, D., et al., J Am Chem Soc, 134: 6947-6951 (2012), and Belliveau, NM, et al., Molecular Therapy-Nucleic Acids, 1: e37 (2012), the entire disclosures of which are incorporated herein by reference.

[0304] Briefly, cargo, such as cargo described herein, is prepared in a first buffer solution. Other lipid nanoparticle components (such as ionizable lipids, conjugate-linker lipids, cholesterol, and phosphorides) are prepared in a second buffer solution. In some embodiments, the two solutions are introduced into a microfluidic device by a syringe pump. The two solutions are contacted in the microfluidic device to form lipid nanoparticles that encapsulate the cargo.

[0305] The disclosed method of screening lipid nanoparticles is described in International Patent Application No. PCT / US2018 / 058171, which is incorporated herein by reference in its entirety. FIG. 1 illustrates an exemplary mRNA screening system for LNP preparations according to an embodiment of the present disclosure. FIG. 2 illustrates an exemplary siRNA screening system for LNP preparations according to an embodiment of the present disclosure. In some embodiments, the screening method characterizes the vehicle delivery preparations to identify preparations that have the desired tropism and deliver functional cargo to the cytoplasm of a particular cell. In some embodiments, the screening method uses a reporter that has functionality that can be detected when delivered to a cell. For example, detecting a functional reporter in the cell indicates that the LNP preparation delivers functional cargo to the cell. Notably, in some embodiments, each different delivery vehicle formulation includes a chemical composition identifier that keeps track of the chemical composition specific to each different delivery vehicle formulation. In some embodiments, the chemical composition identifier is a nucleic acid barcode. In some embodiments, the sequence of the nucleic acid barcode is paired with the chemical components used to formulate the loaded LNP preparation, so that the sequence of the nucleic acid barcode identifies the chemical composition of the delivery vehicle that delivered the barcode. Representative barcodes include, but are not limited to, those described in Sago, 2018 PNAS, Sago, JACS 2018, the entire disclosures of which are incorporated herein by reference. Representative reporters include, but are not limited to, siRNA, mRNA, nuclease protein, nuclease mRNA, small molecules, epigenetic modifiers, and phenotypic modifiers. DNA (genome and DNA barcode) can be isolated using QuickExtract (Lucigen) and sequenced using Illumina MiniSeq as described by Sago et al. PNAS 2018, Sago et al. JACs 2018, Sago, Lokugamage et al. Nano Letters 2018, the entire disclosures of which are incorporated herein by reference.

[0306] V. How to Use Among other things, the present disclosure describes methods of using the compositions, preparations, nanoparticles, and / or nanomaterials described herein. For example, in some embodiments, the present disclosure describes methods of using the compositions, preparations, nanoparticles, and / or nanomaterials described herein to deliver cargo to specific cells, tissues, or organs. As another example, in some embodiments, the present disclosure describes methods of treating and / or slowing and / or halting the progression of a disease or disorder using the compositions, preparations, nanoparticles, and / or nanomaterials described herein. In some embodiments, the compositions, preparations, nanoparticles, and / or nanomaterials described herein are for use in medicine.

[0307] In some embodiments, the compositions, preparations, nanoparticles, and / or nanomaterials described herein deliver a therapeutic or prophylactic agent to a specific cell or organ in a subject in need thereof. In some embodiments, the compositions, preparations, nanoparticles, and / or nanomaterials deliver a therapeutic or prophylactic agent to a specific cell or organ in a subject in need thereof in the absence of a targeting ligand. In some embodiments, the compositions, preparations, nanoparticles, and / or nanomaterials are useful for treating or preventing a disease in a subject in need thereof.

[0308] A. Methods for Delivering Cargo to Cells, Tissues, or Organs In particular, in some embodiments, the compositions, preparations, nanoparticles, and / or nanomaterials disclosed herein are targeted to a particular type or class of cells (e.g., cells of a particular organ or system), tissue, and / or organ. In some embodiments, the disclosure provides methods of delivering one or more cargoes described herein to a subject in need thereof. In some embodiments, such methods include in vivo and / or in vitro delivery. In some embodiments, such methods include in vivo delivery. In some embodiments, such methods include in vitro delivery. In some embodiments, the disclosure provides methods of delivering one or more therapeutic and / or prophylactic nucleic acids to a subject in need thereof, as described herein.

[0309] In some embodiments, the compositions, preparations, nanoparticles, and / or nanomaterials comprise a therapeutic and / or prophylactic agent of interest that can be delivered specifically to liver cells in a subject. Exemplary liver cells include, but are not limited to, hepatocytes.

[0310] In some embodiments, the compositions, preparations, nanoparticles, and / or nanomaterials comprise a therapeutic and / or prophylactic agent of interest that can be delivered specifically to spleen cells in a subject. Exemplary spleen cells include, but are not limited to, spleen monocytes, spleen T cells, spleen memory B cells, or spleen B cells.

[0311] In some embodiments, the compositions, preparations, nanoparticles, and / or nanomaterials comprise a therapeutic and / or prophylactic agent of interest that can be delivered specifically to bone marrow cells in a subject. Exemplary bone marrow cells include, but are not limited to, bone marrow monocytes, bone marrow B cells, bone marrow memory B cells, or bone marrow T cells.

[0312] In some embodiments, the compositions, preparations, nanoparticles, and / or nanomaterials comprise a therapeutic and / or prophylactic agent of interest that can be delivered specifically to immune cells in a subject. Exemplary immune cells include, but are not limited to, CD8+, CD4+, or CD8+CD4+ cells.

[0313] In some embodiments, the compositions, preparations, nanoparticles, and / or nanomaterials comprise a therapeutic and / or prophylactic agent of interest that can be delivered specifically to hematopoietic stem cells in a subject.Unless otherwise specified, it is understood that the terms "hematopoietic stem cells (HSCs)" and "hematopoietic stem and progenitor cells (HSPCs)" are used interchangeably in this disclosure.

[0314] In some embodiments, the compositions, preparations, nanoparticles, and / or nanomaterials comprise a therapeutic and / or prophylactic agent of interest that can be delivered specifically to cardiac cells (e.g., cardiomyocytes).

[0315] In some embodiments, the compositions, preparations, nanoparticles, and / or nanomaterials comprise a therapeutic and / or prophylactic agent of interest that can be delivered specifically to muscle cells (e.g., muscle cells) in a subject.

[0316] In some embodiments, lipid nanoparticles can be formulated to be delivered to mammalian liver hepatocytes, liver immune cells, splenic T cells, or lung endothelial cells in the absence of targeting ligand. Specific delivery to a particular class or type of cell indicates that a higher percentage of lipid nanoparticles are delivered to the target type or class of cells. In some embodiments, specific delivery can result in 2-fold, 5-fold, 10-fold, 15-fold, or more than 20-fold compared to delivery using conventional nanoparticle systems (e.g., MC3-containing LNP).

[0317] B. Methods for Producing Polypeptides In particular, in some embodiments, the methods of using the compositions, preparations, nanoparticles, and / or nanomaterials disclosed herein are used in methods of producing polypeptides. In particular, in some embodiments, the lipid nanoparticles described herein can be used to produce polypeptides in target cells in subjects in need of such production. For example, in some embodiments, the lipid nanoparticles described herein can be used to produce polypeptides in target cells in subjects in need of such production. In some embodiments, the compositions, preparations, nanoparticles, and / or nanomaterials disclosed herein comprise one or more nuclear sequences that are delivered to cells.

[0318] In some embodiments, one or more nucleic acids are expressed in a cell. In some embodiments, expression of a nucleic acid sequence involves one or more of the following: (1) production of an RNA template from a DNA sequence (e.g., by transcription), (2) processing of the RNA transcript (e.g., by splicing, editing, 5' capping, and / or 3' end formation), (3) translation of the RNA into a polypeptide or protein, and / or (4) post-translational modification of the polypeptide or protein.

[0319] C. Methods of Gene Regulation In particular, in some embodiments, the methods of using the compositions, preparations, nanoparticles, and / or nanomaterials disclosed herein are used for gene regulation. In particular, in some embodiments, the lipid nanoparticles described herein can be used to reduce and / or increase gene expression in target cells in a subject in need thereof. For example, in some embodiments, the lipid nanoparticles described herein can deliver one or more nucleic acids to a target cell in a subject without a targeting ligand. In some embodiments, the nucleic acid is an inhibitor nucleic acid. In some embodiments, the inhibitory nucleic acid is an siRNA. In some embodiments, the nucleic acid is a nucleic acid described herein. As another example, in some embodiments, the lipid nanoparticles described herein can deliver a cargo to a target cell in a subject without a targeting ligand. In some embodiments, the cargo is any cargo described herein.

[0320] In particular, in some embodiments, methods of using the compositions, preparations, nanoparticles, and / or nanomaterials disclosed herein to edit genes in cells in a subject in need thereof.

[0321] In some embodiments, the cell targeted for gene regulation is an immune cell. The immune cell can be a T cell, such as a CD8+ T cell, a CD4+ T cell, or a T regulatory cell. Other exemplary immune cells for gene editing include, but are not limited to, macrophages, dendritic cells, B cells, or natural killer cells. In some embodiments, the cell targeted for gene regulation is a hepatocyte.

[0322] Exemplary genes that may be targeted include, but are not limited to, T cell receptor, B cell receptor, CTLA4, PD1, FOXO1, FOXO3, AKTs, CCR5, CXCR4, LAG3, TIM3, killer immunoglobulin-like receptor, GITR, BTLA, LFA-4, T4, LFA-1, Bp35, CD27L receptor, TNFRSF8, TNFRSF5, CD47, CD52, ICAM-1, LFA-3, L-selectin, Ki-24, MB1, B7, B70, M-CSFR, TNFR-II, IL-7 R, OX-40, CD137, CD137L, CD30L, CD40L, FasL, TRAIL, CD257, LIGHT, TRAIL-R1, TRAILR2, TRAIL-R4, TWEAK-R, TNFR, BCMA, B7DC, BTLA, B7-H1, B7-H2, B7-H3, ICOS, VEGFR2, NKG2D, JAG1, GITR, CD4, CCR2, GATA-3, MTORC1, MTORC2, RAPTOR, GATOR, FOXP3, NFAT, IL2R, and IL7. Other exemplary genes that may be targeted include, but are not limited to, OCT, G6Pase, Mut, PCCA, PCCB, and PAH. Exemplary tumor-associated antigens recognized by T cells and contemplated for targeting include, but are not limited to, MAGE1, MAGE3, MAGE6, BAGE, GAGE, NYESO-1, MART1 / Melan A, MC1R, GP100, tyrosinase, TRP-1, TRP-2, PSA, CEA, Cyp-B, Her2 / Neu, hTERT, MUC1, PRAME, WT1, RAS, CDK-4, MUM-1, KRAS, MSLN, and β-catenin.

[0323] D. Subjects to be Treated In some embodiments, the subject being treated is a mammal experiencing cancer, an autoimmune disease, an infectious disease, an organ transplant, an organ failure, a protein deficiency, or a combination thereof. In some embodiments, the subject is a human. In some embodiments, the methods described herein can cause liver cells to translate certain proteins. In some embodiments, the methods described herein can be used to deliver one or more DNA, mRNA, sgRNA, or siRNA to liver cells. In some embodiments, the methods described herein can be used to deliver one or more DNA, mRNA, sgRNA, or siRNA to splenic T cells. In some embodiments, the methods described herein can be used to deliver one or more DNA, mRNA, sgRNA, or siRNA to splenic B cells. In some embodiments, the methods described herein can be used to deliver one or more DNA, mRNA, sgRNA, or siRNA to splenic monocytes. In some embodiments, the methods described herein can be used to deliver one or more DNA, mRNA, sgRNA, or siRNA to bone marrow cells.

[0324] It should be understood that the order or sequence of steps for performing certain operations is immaterial so long as the invention remains operable. Moreover, two or more steps or operations may be performed simultaneously.

[0325] While the present invention has been particularly shown and described with reference to certain preferred embodiments, it should be understood by those skilled in the art that various changes in form and detail may be made therein without departing from the spirit and scope of the invention as defined by the appended claims. Exemplary embodiments 1. Compound of Formula I: [ka] or a pharma- ceutically acceptable salt thereof, wherein: Each L 1 and L 1’is independently -C(O)- or -C(O)O-; Each L 2 and L 2’ are independently a covalent bond, an optionally substituted divalent saturated or unsaturated linear or branched C 1 -C 12 a hydrocarbon chain, or [ka] and Each Cy A is independently an optionally substituted ring selected from phenylene, or a 3- to 7-membered saturated or partially unsaturated carbocyclene; each m is independently 0, 1, or 2; Each L 3 and L 3’ are independently a covalent bond, —C(O)O—, —OC(O)—, —O—, or —OC(O)O—; Each R 1 and R 1’ is independently an optionally substituted group, in which 1 to 3 methylene units are optionally and independently replaced by -O- or -NR-, and 1 -C 20 Hydrocarbon chains; 3- to 12-membered saturated or partially unsaturated carbocyclic rings; 1-adamantyl; 2-adamantyl; sterolyl; phenyl; and [ka] is an optionally substituted group selected from Each L 4 independently, divalent saturated or unsaturated linear or branched C 1 -C 6 is a hydrocarbon chain, Each A 1 and A 2 is independently an optionally substituted C 1 -C 20 Aliphatic or -L 5 -R 5or Or A 1 and A 2 may be taken together with their intervening atoms to form an optionally substituted ring, [ka] During the ceremony, x is selected from 1 or 2; # is L 4 represents the attachment point to Each L 5 independently, divalent saturated or unsaturated linear or branched C 1 -C 20 a hydrocarbon chain in which one to three methylene units are optionally and independently replaced by -O- or -NR-; Each R 5 is independently an optionally substituted group selected from a 5- to 10-membered aryl ring and a 3- to 8-membered carbocyclic ring; X 1 is a covalent bond, -O-, or -NR-; X 2 is a covalent bond or an optionally substituted divalent saturated or unsaturated linear or branched C 1 -C 12 a hydrocarbon chain in which one to three methylene units are optionally and independently replaced by -O- or -NR-; X 3 is hydrogen or -Cy B and Cy B is an optionally substituted ring selected from 3- to 7-membered saturated or partially unsaturated carbocyclyl; phenyl; 3- to 7-membered saturated or partially unsaturated heterocyclyl having 1-3 heteroatoms independently selected from nitrogen, oxygen, and sulfur; and 5- to 6-membered heteroaryl having 1-3 heteroatoms independently selected from nitrogen, oxygen, and sulfur; Each R is independently hydrogen or optionally substituted C 1 -C 6 is an aliphatic group, However, X3 is hydrogen, R 1 or R 1’ At least one of the [ka] or a pharma- ceutically acceptable salt thereof. 2. Each L 1 and L 1’ is independently -C(O)- or -C(O)O-; Each L 2 and L 2’ are independently a covalent bond, an optionally substituted divalent saturated or unsaturated linear or branched C 1 -C 12 a hydrocarbon chain, or [ka] and Each Cy A is independently an optionally substituted ring selected from phenylene, and a 3- to 7-membered saturated or partially unsaturated carbocyclene; each m is independently 0, 1, or 2; Each L 3 and L 3’ are independently a covalent bond, —C(O)O—, —OC(O)—, —O—, or —OC(O)O—; Each R 1 and R 1’ is independently an optionally substituted group, in which 1 to 3 methylene units are optionally and independently replaced by -O- or -NR-, and 1 -C 20 Hydrocarbon chains; 3- to 7-membered saturated or partially unsaturated carbocyclic rings; 1-adamantyl; 2-adamantyl; sterolyl; phenyl; and [ka] is an optionally substituted group selected from Each L 4 independently, divalent saturated or unsaturated linear or branched C 1 -C 6 is a hydrocarbon chain, Each A 1 and A 2 is independently an optionally substituted C 1 -C 20 Aliphatic or -L 5 -R 5 or Or A 1 and A 2 may be taken together with their intervening atoms to form an optionally substituted ring, [ka] During the ceremony, x is selected from 1 or 2; # is L 4 represents the attachment point to Each L 5 independently, divalent saturated or unsaturated linear or branched C 1 -C 20 a hydrocarbon chain in which one to three methylene units are optionally and independently replaced by -O- or -NR-; Each R 5 is independently an optionally substituted group selected from a 5- to 10-membered aryl ring and a 3- to 8-membered carbocyclic ring; X 1 is a covalent bond, -O-, or -NR-; X 2 is a covalent bond or an optionally substituted divalent saturated or unsaturated linear or branched C 1 -C 12 a hydrocarbon chain in which one to three methylene units are optionally and independently replaced by -O- or -NR-; X 3 is hydrogen or -Cy B and Cy Bis an optionally substituted ring selected from 3- to 7-membered saturated or partially unsaturated carbocyclyl; phenyl; 3- to 7-membered saturated or partially unsaturated heterocyclyl having 1-3 heteroatoms independently selected from nitrogen, oxygen, and sulfur; and 5- to 6-membered heteroaryl having 1-3 heteroatoms independently selected from nitrogen, oxygen, and sulfur; Each R is independently hydrogen or optionally substituted C 1 -C 6 is an aliphatic group, However, X 3 is hydrogen, R 1 or R 1’ At least one of the [ka] 2. The compound of embodiment 1, wherein 3. The compound is a compound of formula (II): [ka] or a pharma- ceutically acceptable salt thereof. 4. Cy B is an optionally substituted 3-7 membered saturated or partially unsaturated heterocyclyl having 1-3 heteroatoms independently selected from nitrogen, oxygen, and sulfur. 5. Cy B is optionally substituted pyrrolidinyl or piperidinyl; Cy B The carbon atoms in X 2 The compound of any one of embodiments 1 to 4, wherein 6. Cy B But, C 1 -C 6 The compound according to any one of embodiments 1 to 5, which is optionally aliphatically substituted. 7. Cy B but, [ka] and an optionally substituted ring selected from C 1 -C 6 The compound according to any one of embodiments 1 to 6, which is optionally aliphatically substituted. 8. Cy B but, [ka] The compound of any one of embodiments 1-7, wherein 9. Cy B but, [ka] The compound of any one of embodiments 1-7, wherein 10.Each R 1 and R 1’ is independently an optionally substituted group, in which 1 to 3 methylene units are optionally and independently replaced by -O- or -NR-, and 1 -C 20 The compound of any one of embodiments 1-9, wherein R is an optionally substituted group selected from: a hydrocarbon chain; a 3-7 membered saturated or partially unsaturated carbocyclic ring; 1-adamantyl; 2-adamantyl; sterolyl; and phenyl. 11 each R 1 and R 1’ But independently, [ka] 1-adamantyl, [ka] The compound according to any one of embodiments 1 to 10, selected from: 12.Each R 1 and R 1’ but, [ka] 1-adamantyl, [ka] The compound of any one of embodiments 1-10, independently selected from: 13. The compound is a compound of formula (III): [ka] or a pharma- ceutically acceptable salt thereof. 14.L 1 and L 1’ The compound of any one of embodiments 1-13, wherein is -C(O)-. 15. The compound is a compound of formula (IV-A) or (IV-B): [ka] or a pharma- ceutically acceptable salt thereof. 16.R 1 is an optionally substituted group, in which 1 to 3 methylene units are optionally and independently replaced by -O- or -NR-, and 1 -C 20 The compound of any one of embodiments 13-15, wherein R is an optionally substituted group selected from: a hydrocarbon chain; a 3-7 membered saturated or partially unsaturated carbocyclic ring; 1-adamantyl; 2-adamantyl; sterolyl; and phenyl. 17.R 1’ but, [ka] 1-adamantyl, [ka] 17. The compound according to any one of embodiments 13 to 16, selected from: 18.R 1’ but, [ka] 1-adamantyl, [ka] 17. The compound according to any one of embodiments 13 to 16, selected from: 19. The compound is a compound of formula (VA) or (VB): [ka] or a pharma- ceutically acceptable salt thereof. 20.X 1 The compound of any one of embodiments 1-19, wherein is -NR-. 21.X 1 The compound according to any one of embodiments 1 to 20, wherein is -NH-. 22.X 1 The compound of any one of embodiments 1-19, wherein is -O-. 23.X 2 optionally substituted divalent saturated or unsaturated linear or branched C 1 -C 12 The compound according to any one of embodiments 1 to 22, which is a hydrocarbon chain. 24.X 2 The compound of any one of embodiments 1-23, wherein is methylene, ethylene, or propylene. 25.Each L 2 and L 2’ are independently a covalent bond, an optionally substituted divalent saturated or unsaturated linear or branched C 1 -C 6 a hydrocarbon chain, or [ka] The compound of any one of embodiments 1-24, wherein 26. Cy A is cyclohexylene or phenylene. 27.Each L 2 and L 2’ is independently a covalent bond, methylene, ethylene, propylene, butylene, pentylene, or hexylene. 28.L 2 and L 2’ The compound of any one of embodiments 1-24, wherein is a covalent bond. 29.Each L 3 and L 3’ The compound of any one of embodiments 1-28, wherein is independently a covalent bond or -C(O)O-. 30.L 3 and L 3’ The compound of any one of embodiments 1-29, wherein is a covalent bond. 31.L 3 is a covalent bond, and L 3’ The compound of any one of embodiments 1-29, wherein is -C(O)O-. 32.Each L 4 is independently methylene, ethylene, or propylene. 33.Each A 1 and A 2 , optionally substituted C 1 -C 20 The compound of any one of embodiments 1-9 or 13-32, independently selected from aliphatic. 34.Each A 1 and A 2 , optionally substituted C 8 The compound of any one of embodimen...

Claims

1. Compounds of Formula I: 【Chemistry 1】 or a pharma- ceutically acceptable salt thereof, wherein: Each L １ and L １’ is independently —C(O)— or —C(O)O—; Each L ２ and L ２’ are independently a covalent bond, an optionally substituted divalent saturated or unsaturated linear or branched C １ -C １２ a hydrocarbon chain, or 【Chemistry 2】 and Each Cy Ａ is independently an optionally substituted ring selected from phenylene, and 3- to 7-membered saturated or partially unsaturated carbocyclene; each m is independently 0, 1, or 2; Each L ３ and L ３’ are independently a covalent bond, —C(O)O—, —OC(O)—, —O—, or —OC(O)O—; Each R １ and R １’ is independently an optionally substituted group, in which 1 to 3 methylene units are optionally and independently replaced by -O- or -NR-, and １ -C ２０ Hydrocarbon chains; 3-12 membered saturated or partially unsaturated carbocyclic rings; 1-adamantyl; 2-adamantyl; sterolyl; Phenyl; and 【Chemistry 3】 is an optionally substituted group selected from Each L ４ independently, a divalent saturated or unsaturated linear or branched C １ -C ６ is a hydrocarbon chain, Each A １ and A ２ is independently an optionally substituted C １ -C ２０ Aliphatic or -L ５ -R ５ or Or A １ and A ２ may be taken together with their intervening atoms to form an optionally substituted ring: 【Chemistry 4】 During the ceremony, x is selected from 1 or 2; # is L ４ represents the attachment point to Each L ５ independently, a divalent saturated or unsaturated linear or branched C １ -C ２０ a hydrocarbon chain in which one to three methylene units are optionally and independently replaced by -O- or -NR-; Each R ５ is independently an optionally substituted group selected from a 5- to 10-membered aryl ring and a 3- to 8-membered carbocyclic ring; X １ is a covalent bond, —O—, or —NR—; X ２ is a covalent bond or an optionally substituted divalent saturated or unsaturated linear or branched C １ -C １２ a hydrocarbon chain in which one to three methylene units are optionally and independently replaced by -O- or -NR-; X ３ is hydrogen or -Cy Ｂ and Cy Ｂ is an optionally substituted ring selected from 3- to 7-membered saturated or partially unsaturated carbocyclyl; phenyl; 3- to 7-membered saturated or partially unsaturated heterocyclyl having 1-3 heteroatoms independently selected from nitrogen, oxygen, and sulfur; and 5- to 6-membered heteroaryl having 1-3 heteroatoms independently selected from nitrogen, oxygen, and sulfur; Each R is independently hydrogen or optionally substituted C １ -C ６ is an aliphatic group, However, X ３ is hydrogen, R １ Or R １’ At least one of 【Chemistry 5】 or a pharma- ceutically acceptable salt thereof.

2. The compound is a compound of formula (II), (III), (IV-A), (IV-B), (VA), or (V-B): 【Chemistry 6】 【Chemistry 9】 【Chemistry 10】 【Chemistry 13】 2. The compound of claim 1, or a pharma- ceutically acceptable salt thereof.

3. Each R １ and R １’ is independently an optionally substituted group, in which 1 to 3 methylene units are optionally and independently replaced by -O- or -NR-, and １ -C ２０ an optionally substituted group selected from: a hydrocarbon chain; a 3- to 7-membered saturated or partially unsaturated carbocyclic ring; 1-adamantyl; 2-adamantyl; sterolyl; and phenyl; or Each R 1 and R 1′ is 【Chemistry 7】 1-adamantyl, 【Chemistry 8】 are independently selected from The compound is a compound of formula (IV-A) or (IV-B): 【Chemistry 10】 and R 1 is an optionally substituted group selected from a saturated or unsaturated, linear or branched C 1 -C 20 hydrocarbon chain in which one to three methylene units are optionally and independently replaced by -O- or -NR-; a 3- to 7-membered saturated or partially unsaturated carbocyclic ring; 1-adamantyl; 2-adamantyl; sterolyl; phenyl; or R 1' is 【Chemistry 11】 1-adamantyl, 【Chemistry 12】 Selected from:

2. The compound of claim 1 or a pharma- ceutically acceptable salt thereof.

4. L １ and L １’ is —C(O)—, and / or Each L 2 and L 2′ is independently a covalent bond, an optionally substituted divalent saturated or unsaturated straight or branched C 1 -C 6 hydrocarbon chain, or 【Chemistry 14】 and in particular Cy A is cyclohexylene or phenylene, or each L 2 and L 2′ is independently a covalent bond, methylene, ethylene, propylene, butylene, pentylene, or hexylene, in particular L 2 and L 2′ are a covalent bond; and / or each L 3 and L 3' is independently a covalent bond or -C(O)O, in particular L 3 and L 3' are a covalent bond or L 3 is a covalent bond and L 3' is -C(O)O; The compound of claim 1.

5. X １ is -NR-, in particular -NH-, or X １ is -O-, and / or X 2 is an optionally substituted divalent saturated or unsaturated straight or branched C 1 -, C 12 hydrocarbon chain, in particular X 2 is methylene, ethylene or propylene; The compound of claim 1.

6. Each L ４ The compound of claim 1 , wherein is independently methylene, ethylene, or propylene.

7. Each A １ and A ２ is optionally substituted C １ -C ２０ independently selected from aliphatic, or each A 1 and A 2 is independently selected from optionally substituted C 8 aliphatic; or Each of A 1 and A 2 is 【Chemistry 15】 and / or each A 1 and A 2 is the same; The compound of claim 1.

8. Cy Ｂ is an optionally substituted 3-7 membered saturated or partially unsaturated heterocyclyl having 1-3 heteroatoms independently selected from nitrogen, oxygen, and sulfur, in particular Cy B is an optionally substituted pyrrolidinyl or piperidinyl, and a carbon atom within Cy B is bonded to X 2 , or Cy B is optionally substituted with C 1 -C 6 aliphatic; or Cy B is 【Chemistry 16】 and optionally C 1 -C 6 aliphatically substituted, particularly when Cy B is 【Chemistry 17】 or 【Chemistry 18】 That is, The compound of claim 1.

9. The compound is 【Chemistry 19】 【change】 【change】 【change】 【change】 【change】 【change】 【change】 【change】 2. The compound of claim 1, selected from:

10. A lipid nanoparticle (LNP) preparation comprising an ionizable lipid according to any one of claims 1 to 9, optionally further comprising Phospholipids and Cholesterol and A conjugate-linker lipid (e.g., a polyethylene glycol lipid), and an LNP preparation.

11. and / or a therapeutic and / or prophylactic agent, in particular the therapeutic and / or prophylactic agent is or comprises one or more nucleic acids, in particular the one or more nucleic acids is or comprises RNA or DNA, and / or The LNP preparations are formulated to deliver the therapeutic and / or prophylactic agent to target cells, particularly where the target cells are or comprise spleen cells (e.g., splenic B cells, splenic T cells, splenic monocytes), liver cells (e.g., hepatocytes), bone marrow cells (e.g., bone marrow monocytes), immune cells, muscle cells (e.g., myoocytes), cardiac cells (e.g., cardiomyocytes), kidney cells, or cells in the central nervous system, or where the target cells are or comprise hematopoietic stem cells. The LNP preparation of claim 10.

12. A pharmaceutical composition comprising the LNP preparation of claim 10 and a pharma- ceutically acceptable excipient.

13. The LNP preparation of claim 10 for use in a method comprising: The method comprises: A method for administering a therapeutic and / or prophylactic agent to a subject in need thereof, comprising administering to said subject the LNP preparation of claim 10; or A method for treating a disease or disorder in a subject in need thereof, the method comprising administering to the subject an LNP preparation according to claim 10, wherein the therapeutic and / or prophylactic agent is effective in treating the disease or A method for slowing and / or halting the progression of a disease or disorder in a subject in need thereof, said method comprising administering to said subject the LNP preparation of claim 10, wherein said therapeutic and / or prophylactic agent is effective in treating said disease or 11. A method of producing a polypeptide of interest in a mammalian cell, said method comprising contacting said cell with the LNP preparation of claim 10, wherein said therapeutic and / or prophylactic agent is or comprises an mRNA, said mRNA encoding said polypeptide of interest, whereby said mRNA is capable of being translated in said cell to produce said polypeptide of interest, or 11. A method of inhibiting production of a polypeptide of interest in a mammalian cell, the method comprising contacting the cell with the LNP preparation of claim 10, wherein the therapeutic and / or prophylactic agent is or comprises an RNA, whereby the RNA is capable of inhibiting production of the polypeptide of interest, or A method for specifically delivering a therapeutic and / or prophylactic agent to a mammalian organ, the method comprising contacting a mammalian organ with the LNP preparation of claim 10, whereby the therapeutic and / or prophylactic agent is delivered to the organ, in particular comprising administering to a subject the LNP preparation of claim 10; or A method of vaccination by administering the LNP preparation of claim 10. LNP preparation.

14. A method for delivering a therapeutic and / or prophylactic agent to mammalian cells derived from a subject, comprising contacting the cells of the subject that has been administered the LNP preparation of claim 10.

15. An LNP preparation comprising a compound according to any one of claims 1 to 9 for use in a method of inducing an adaptive immune response in a subject, the method comprising administering to the subject an effective amount of a composition comprising at least one RNA, the composition comprising a compound according to any one of claims 1 to 9, or a pharma- ceutically acceptable salt thereof.

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