Ionizable lipids with degradable head groups

Abstract

A compound having the following structure of formula (I), or a stereoisomer, salt, or tautomer thereof, wherein R 1 , R 2 , R 3 , R 4 , R 5 , R 6 , R 7 , R 8 , X, Y 1 , and Y 2 or a stereoisomer, salt, or tautomer thereof, wherein: is as defined herein. Pharmaceutical compositions and lipid nanoparticles comprising these compounds, and their use in methods of treating disease, are also described. [Case 1] JPEG2026502350000052.jpg44170

Description

[Technical Field]

[0001] CROSS-REFERENCE TO RELATED APPLICATIONS This application claims priority to U.S. Provisional Patent Application No. 63 / 476,993, filed December 23, 2022, the contents of which are incorporated herein by reference in their entirety.

[0002] The present disclosure relates to ionizable lipids that can be used alone or in combination with other lipid components, such as helper lipids, stabilizing lipids, and / or structural lipids. The present disclosure also provides lipid-nanoparticle compositions comprising such lipids for the delivery of molecules, particularly nucleic acids. [Background technology]

[0003] Nucleic acid-based therapy has attracted attention in recent years because it has great potential to treat diseases by targeting their genetic blueprint in vivo.Nucleic acid-based therapeutic agents can achieve long-term or even curative effects through gene inhibition, addition, replacement, or editing.However, the clinical translation of both nucleic acid drugs and other therapeutic molecules depends on delivery technologies that improve stability, promote internalization, and / or increase target affinity.

[0004] Lipid-based delivery systems, such as but not limited to lipid nanoparticles (LNPs), can provide an approach for stabilizing and delivering nucleic acids and other therapeutic molecules, and there remains a significant need for advances in this technology. Further advances in design characteristics, such as optimal particle size, encapsulation efficiency, robust manufacturing processes, different lipid affinities, and neutral surface charge, can provide efficient lipid-based delivery systems for nucleic acids and other therapeutic molecules. Summary of the Invention

[0005] In summary, the present disclosure provides compounds, including their stereoisomers, pharmaceutically acceptable salts, or tautomers, that can be used alone or in combination with other therapeutic agents.

[0006] In one embodiment, a compound is provided having the structure of Formula (I), or a stereoisomer, salt, or tautomer thereof, wherein R 1 , R 2 , R 3 , R 4 , X, Y 1 , and Y 2 is as defined herein. [ka]

[0007] Also provided are pharmaceutical compositions comprising one or more of the compounds of formula (I) above and a therapeutic agent.

[0008] In other embodiments, methods of treatment are provided by administering to a subject in need thereof a compound of formula (I) as described above, or a pharmaceutical composition comprising a compound of formula (I), to treat a disease.

[0009] In one embodiment, lipid nanoparticles are also provided that comprise one or more of the compounds of formula (I) above and a therapeutic agent that comprises a nucleic acid.

[0010] Various aspects and embodiments are now more fully described below. Such aspects and embodiments may take many different forms, and the illustrative examples disclosed herein are not intended to be limiting; rather, these embodiments are provided so that this disclosure will be thorough and complete, and will fully convey its scope to those skilled in the art. DETAILED DESCRIPTION OF THE INVENTION

[0011] I. Definition For convenience, certain terms used in the specification and claims are collected here. Unless defined otherwise, all technical and scientific terms used in this disclosure have the same meaning as commonly understood by one of ordinary skill in the art to which this disclosure belongs.

[0012] When a range of values ​​is provided, each intervening value between the upper and lower limits of that range, and any other stated or intervening value within that stated range, is intended to be encompassed within the disclosure. For example, if a range of 1 μm to 8 μm is stated, 2 μm, 3 μm, 4 μm, 5 μm, 6 μm, and 7 μm, as well as ranges of values ​​above 1 μm and below 8 μm, are also intended to be expressly disclosed.

[0013] Unless the context clearly dictates otherwise, the singular forms "a," "an," and "the" include plural referents. Thus, for example, reference to a "polymer" includes a single polymer as well as two or more of the same or different polymers, reference to an "excipient" includes a single excipient as well as two or more of the same or different excipients, etc.

[0014] The term "about" is intended to encompass a deviation of plus or minus 5 percent, particularly with respect to a given quantity.

[0015] The compositions of the present disclosure can comprise, consist essentially of, or consist of the disclosed components.

[0016] All percentages, amounts, and ratios are based on the total weight of the composition unless otherwise specified, and all measurements made are at about 25°C.

[0017] As used in the specification and appended claims, unless specified to the contrary, the following terms have the meanings indicated. "Amino" refers to the radical -NH2, -NHR, or -NR2. "Cyano" refers to the -CN radical. "Hydroxyl" refers to the -OH radical. "Imino" refers to a =NH or =NR substituent. "Nitro" refers to the -NO2 radical. "Oxo" refers to the =O substituent. "Thio" refers to the ═S substituent. "Trifluoromethyl" refers to the -CF3 radical. Hylazide or hydrazino refers to an N-N substituent. wherein each R is a compatible substituent as described in this disclosure. When an R group is chiral, isomers are contemplated and included herein.

[0018] "Alkyl" refers to a straight-chain, saturated, acyclic, monovalent hydrocarbon radical or a branched, saturated, acyclic, monovalent hydrocarbon radical having 1 to 12 carbon atoms, preferably 1 to 8 carbon atoms, or 1 to 6 carbon atoms, attached to the rest of the molecule by a single bond, e.g., methyl, ethyl, n-propyl, 1-methylethyl (iso-propyl), n-butyl, n-pentyl, 1,1-dimethylethyl (t-butyl), 3-methylhexyl, 2-methylhexyl, and the like. Optionally substituted alkyl radicals, where valence allows, independently include halo, cyano, nitro, aryl, cycloalkyl, heterocyclyl, heteroaryl, oxo, trimethylsilanyl, —OR′, —OC(O)—R′, —N(R′)2, —C(O)R″, —C(O)OR′, —C(O)N(R′)2, —N(R′)C(O)OR′″, —N(R′)C(O)R′″, —N(R′)S(O) t R′″ (wherein t is 1 or 2), —S(O) t OR′′′ (wherein t is 1 or 2), —S(O) p R′′′ (wherein p is 0, 1, or 2), and —S(O) tN(R')2, where t is 1 or 2, wherein each R' is independently hydrogen, alkyl, haloalkyl, cycloalkyl, cycloalkylalkyl, aryl, aralkyl, heterocyclyl, or heteroaryl; each R" is independently hydrogen, cycloalkyl, aryl, heterocyclyl, or heteroaryl; and each R'" is independently alkyl, haloalkyl, cycloalkyl, aryl, heterocyclyl, or heteroaryl.

[0019] "Alkoxy" means a group of the formula -OR a where R a is an alkyl radical as defined above containing 1 to 12 carbon atoms. The alkyl portion of an optionally substituted alkoxy radical is optionally substituted as defined above for an alkyl radical.

[0020] "Alkoxyalkyl" refers to a group of the formula -R a -OR b where R a is alkylene and R b is alkyl as defined above. The alkyl and alkylene portions of an optionally substituted alkoxyalkyl radical are optionally substituted as defined above for the alkyl radical and alkylene chain, respectively.

[0021] "Aralkyl" is a group of the formula -R a -R b where R a is alkylene and R b is aryl as described herein. The alkylene and aryl portions of an optionally substituted aralkyl are optionally substituted as described herein for alkylene and aryl, respectively.

[0022] "Aryl" refers to an aromatic monocyclic or polycyclic hydrocarbon ring system radical containing 6 to 18 carbon atoms; polycyclic aryl ring systems are bicyclic, tricyclic, or tetracyclic ring systems. Aryl radicals include, but are not limited to, groups such as fluorenyl, phenyl, and naphthyl. Optionally substituted aryl is independently selected from alkyl, alkenyl, halo, haloalkyl, haloalkenyl, cyano, nitro, aryl, heteroaryl, heteroarylalkyl, -R"-OR", -R"-OC(O)-R", -R"-N(R)2, -R"-C(O)R", -R"-C(O)OR", -R"-C(O)N(R)2, -R"-N(R)C(O)OR"", -R"-N(R)C(O)R"", -R"-N(R)S(O) t R′′′ (wherein t is 1 or 2), —R′′—S(O) t OR′′′ (wherein t is 1 or 2), —R′′—S(O) p R′′′ (wherein p is 0, 1, or 2), and —R′′—S(O) t N(R')2, where t is 1 or 2, wherein each R' is independently hydrogen, alkyl, haloalkyl, cycloalkyl, aryl, heterocyclyl, or heteroaryl; each R" is independently a direct bond or a straight or branched alkylene or alkenylene chain; and each R'" is independently alkyl, haloalkyl, cycloalkyl, cycloalkylalkyl, aryl, heterocyclyl, or heteroaryl.

[0023] "Arylalkoxy" refers to a group of formula -OR, where R is aralkyl. An optionally substituted arylalkoxy is an arylalkoxy optionally substituted as described herein for aralkyl. In some embodiments, the arylalkoxy is benzyloxy.

[0024] "Cycloalkyl" refers to a stable non-aromatic monocyclic or polycyclic hydrocarbon radical having 3 to 15 carbon atoms, preferably 3 to 10 carbon atoms, saturated or unsaturated, and attached to the rest of the molecule by a single bond. Polycyclic hydrocarbon radicals are bicyclic, tricyclic, or tetracyclic ring systems. Unsaturated cycloalkyls contain one, two, or three carbon-carbon double bonds and / or one carbon-carbon triple bond. Monocyclic cycloalkyl radicals include, for example, cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, cycloheptyl, and cyclooctyl. Polycyclic cycloalkyl radicals include, for example, adamantyl, norbornyl, decalinyl, and the like. Optionally substituted cycloalkyl is independently selected from alkyl, alkenyl, halo, haloalkyl, haloalkenyl, cyano, nitro, oxo, aryl, aralkyl, cycloalkyl, heterocyclyl, heteroaryl, -R''-OR'', -R''-OC(O)-R'', -R''-N(R''), -R''-C(O)R'', -R''-C(O)OR'', -R''-C(O)N(R''), -R''-N(R'')C(O)OR'''', -R''-N(R'')C(O)R'''', -R''-N(R'')S(O) t R′′′ (wherein t is 1 or 2), —R′′—S(O) t OR′′′ (wherein t is 1 or 2), —R′′—S(O) p R′′′ (wherein p is 0, 1, or 2), and —R′′—S(O) t N(R')2, where t is 1 or 2, where each R' is independently hydrogen, alkyl, haloalkyl, cycloalkyl, aryl, heterocyclyl, or heteroaryl; each R" is independently a direct bond or a straight or branched alkylene or alkenylene chain; and each R'" is independently alkyl, haloalkyl, cycloalkyl, cycloalkylalkyl, aryl, aralkyl, heterocyclyl, or heteroaryl.

[0025] A "deuterated compound" is a compound in which one of the more hydrogen atoms has been replaced with a deuterium atom. A deuterated drug may be a derivative of an active compound. A deuterated drug may be a prodrug. Deuteration may alter the physical properties, metabolic properties, activity, or safety of a drug.

[0026] "Derivatives" are related chemical species that can be derived from similar compounds through chemical reaction. They can involve slight chemical modifications, replacement of atoms with deuterated atoms, replacement of atoms with stable or radioactive isotopes, or other modifications that impart desirable properties to the compound.

[0027] "Fused" refers to any ring system described herein that is fused to an existing ring structure in a compound. When the fused ring system is a heterocyclyl or heteroaryl, any carbon atom on the existing ring structure that becomes part of the fused ring system can be replaced with a nitrogen atom.

[0028] "Halo" refers to the halogen substituents: bromo, chloro, fluoro, and iodo.

[0029] "Haloalkyl" refers to an alkyl radical, as defined above, further substituted with one or more halogen substituents. The number of halo substituents contained in a haloalkyl is from one to the total number of hydrogen atoms available for replacement with halo substituents (e.g., perfluoroalkyl). Non-limiting examples of haloalkyl include trifluoromethyl, difluoromethyl, trichloromethyl, 2,2,2-trifluoroethyl, 1-fluoromethyl-2-fluoroethyl, 3-bromo-2-fluoropropyl, 1-bromomethyl-2-bromoethyl, and the like. For optionally substituted haloalkyls, the hydrogen atoms bonded to the carbon atoms of the alkyl portion of the haloalkyl radical can be optionally replaced with the substituents defined above for optionally substituted alkyl.

[0030] "Haloalkenyl" refers to an alkenyl radical, as defined above, which is further substituted with one or more halo substituents. The number of halo substituents contained in a haloalkenyl is from one to the total number of hydrogen atoms available for replacement with halo substituents (e.g., perfluoroalkenyl). Non-limiting examples of haloalkenyls include 2,2-difluoroethenyl, 3-chloroprop-1-enyl, and the like. For optionally substituted haloalkenyls, a hydrogen atom bonded to a carbon atom of the alkenyl portion of the haloalkenyl radical can be optionally replaced with a substituent as defined above for an optionally substituted alkenyl group.

[0031] "Haloalkynyl" refers to an alkynyl radical, as defined above, further substituted with one or more halo substituents. The number of halo substituents contained in the haloalkynyl is from one to the total number of hydrogen atoms available for replacement with halo substituents (e.g., perfluoroalkynyl). Non-limiting examples of haloalkynyl include 3-chloroprop-1-ynyl and the like. The alkynyl portion of the haloalkynyl radical can be additionally optionally substituted as defined above for an alkynyl group.

[0032] "Heteroarylalkyl" refers to a group of the formula -R a -R b where R a is alkylene and R b is heteroaryl. The alkylene and heteroaryl portions of an optionally substituted heteroarylalkyl are optionally substituted as described herein for alkylene and heteroaryl, respectively.

[0033] "Heterocyclyl" refers to a stable 3- to 18-membered non-aromatic ring system radical having 2 to 12 carbon atoms and a total of 1 to 6 heteroatoms independently selected from the group consisting of nitrogen, oxygen, phosphorus, and sulfur. Heterocyclyl radicals are monocyclic, bicyclic, tricyclic, or tetracyclic ring systems. Bicyclic, tricyclic, or tetracyclic heterocyclyls are fused, spiro, and / or bridged ring systems. Heterocyclyl radicals can be saturated or unsaturated. Unsaturated heterocyclyls contain one, two, or three carbon-carbon double bonds and / or one carbon-carbon triple bond. Optionally substituted heterocyclyl is independently alkyl, alkenyl, halo, haloalkyl, haloalkenyl, cyano, oxo, thioxo, nitro, aryl, aralkyl, cycloalkyl, heterocyclyl, heteroaryl, -R''-OR'', -R''-OC(O)-R'', -R''-N(R''), -R''-C(O)R'', -R''-C(O)OR'', -R''-C(O)N(R''), -R''-N(R'')C(O)OR'''', -R''-N(R'')C(O)R'''', -R''-N(R'')S(O) t R′′′ (wherein t is 1 or 2), —R′′—S(O) t OR′′′ (wherein t is 1 or 2), —R′′—S(O) p R′′′ (wherein p is 0, 1, or 2), and —R′′—S(O) tN(R')2, where t is 1 or 2, wherein each R' is independently hydrogen, alkyl, alkenyl, haloalkyl, cycloalkyl, aryl, heterocyclyl, or heteroaryl; each R" is independently a direct bond or a straight or branched alkylene or alkenylene chain; and each R'" is independently alkyl, alkenyl, haloalkyl, cycloalkyl, aryl, heterocyclyl, or heteroaryl. The nitrogen, carbon, or sulfur atom in a heterocyclyl radical can be optionally oxidized (when the substituent is oxo and is on a heteroatom), and the nitrogen atom can be optionally quaternized (when the substituent is alkyl, alkenyl, aryl, aralkyl, cycloalkyl, heterocyclyl, heteroaryl, -R''-OR'', -R''-OC(O)-R'', -R''-N(R''), -R''-C(O)R'', -R''-C(O)OR'', -R''-C(O)N(R''), -R''-N(R'')C(O)OR'''', -R''-N(R'')C(O)R'''', -R''-N(R'')S(O) t R′′′ (wherein t is 1 or 2), —R′′—S(O) t OR′′′ (wherein t is 1 or 2), —R′′—S(O) p R′′′ (wherein p is 0, 1, or 2), and —R′′—S(O) tN(R')2, where t is 1 or 2, where R'' is a straight or branched alkylene or alkenylene chain, and R' and R''' are as defined above. Examples of optionally substituted heterocyclyl radicals include, but are not limited to, azetidinyl, dioxolanyl, thienyl[1,3]dithianyl, decahydroisoquinolyl, imidazolinyl, imidazolidinyl, isothiazolidinyl, isoxazolidinyl, morpholinyl, octahydroindolyl, octahydroisoindolyl, 2-oxopiperazinyl, 2-oxopiperidinyl, 2-oxopyrrolidinyl, oxazolidinyl, piperidinyl, piperazinyl, 4-piperidonyl, pyrrolidinyl, pyrazolidinyl, thiazolidinyl, tetrahydrofur, trithianyl, tetrahydropyranyl, thiomorpholinyl, thiamorpholinyl, 1-oxo-thiomorpholinyl, and 1,1-dioxo-thiomorpholinyl.

[0034] "Heterocyclylene" refers to a heterocyclyl in which one hydrogen atom is replaced by a valence. An optionally substituted heterocyclylene is optionally substituted as described herein for heterocyclyl.

[0035] "Heteroaryl" refers to a 5- to 18-membered ring system radical containing at least one aromatic ring, having 1 to 17 carbon atoms, and containing a total of 1 to 10 heteroatoms independently selected from the group consisting of nitrogen, oxygen, and sulfur. Heteroaryl radicals are monocyclic, bicyclic, tricyclic, or tetracyclic ring systems. Bicyclic, tricyclic, or tetracyclic heteroaryl radicals are fused and / or bridged ring systems. Optionally substituted heteroaryl is independently alkyl, alkenyl, alkoxy, halo, haloalkyl, haloalkenyl, cyano, oxo, thioxo, nitro, oxo, aryl, aralkyl, cycloalkyl, heterocyclyl, heteroaryl, or heteroarylalkyl, -R''-OR'', -R''-OC(O)-R'', -R''-N(R''), -R''-C(O)R'', -R''-C(O)OR'', -R''-C(O)N(R''), -R''-N(R'')C(O)OR'''', -R''-N(R'')C(O)R'''', -R''-N(R'')S(O) t R′′′ (wherein t is 1 or 2), —R′′—S(O) t OR′′′ (wherein t is 1 or 2), —R′′—S(O) t R′′′ (wherein p is 0, 1, or 2), and —R′′—S(O) tN(R')2, where t is 1 or 2, where each R' is independently hydrogen, alkyl, alkenyl, haloalkyl, cycloalkyl, aryl, heterocyclyl, or heteroaryl; each R" is independently a direct bond or a straight or branched alkylene or alkenylene chain; and each R'" is alkyl, alkenyl, haloalkyl, cycloalkyl, aryl, heterocyclyl, or heteroaryl. The nitrogen, carbon, or sulfur atom in a heterocyclyl radical may be optionally oxidized (if the substituent is oxo and present on a heteroatom), provided that at least one ring in the heteroaryl remains aromatic, and the nitrogen atom may be optionally quaternized (if the substituent is alkyl, alkenyl, aryl, aralkyl, cycloalkyl, heterocyclyl, heteroaryl, -R''-OR'', -R''-OC(O)-R'', -R''-N(R''), -R''-C(O)R'', -R''-C(O)OR'', -R''-C(O)N(R''), -R''-N(R'')C(O)OR'''', -R''-N(R'')C(O)R'''', -R''-N(R'')S(O) t R′′′ (wherein t is 1 or 2), —R′′—S(O) t OR′′′ (wherein t is 1 or 2), —R′′—S(O) p R′′′ (wherein p is 0, 1, or 2), and —R′′—S(O) tN(R')2, where t is 1 or 2, where R'' is a straight or branched alkylene or alkenylene chain, and R' and R''' are as defined above. Examples of optionally substituted heteroaryl radicals include azepinyl, acridinyl, benzimidazolyl, benzthiazolyl, benzindolyl, benzodioxolyl, benzofuranyl, benzoxazolyl, benzothiazolyl, benzothiadiazolyl, benzo[b][1,4]dioxepinyl, 1,4-benzodioxanyl, benzonaphthofuranyl, benzoxazolyl, benzodioxolyl, benzodioxinyl, benzopyranyl, the benzopyranonyl group benzofuranyl, benzofuranonyl, benzothienyl (benzothiophenyl), benzotriazolyl, benzo[4,6]imidazo[1,2-a]pyridinyl, the carbazolyl group cinnolinyl, dibenzofuranyl, dibenzothiophenyl, furanyl, furanonyl, isothiazolyl, imidazolyl, These include, but are not limited to, indazolyl, indolyl, indazolyl, isoindolyl, indolinyl, isoindolinyl, isoquinolyl, indolizinyl, isoxazolyl, naphthyl, naphthyridinyl, oxadiazolyl, 2-oxoazepinyl, oxazolyl, oxiranyl, 1-phenyl-1H-pyrrolyl, phenazinyl, phenothiazinyl, phenoxazinyl, phthalazinyl, pteridinyl, purinyl, pyrrolyl, pyrazolyl, pyridinyl, pyrazinyl, pyrimidinyl, pyridazinyl, pyrrolyl, quinazolinyl, quinoxalinyl, quinolinyl, quinuclidinyl, isoquinolinyl, tetrahydroquinolinyl, thiazolyl, thiadiazolyl, triazolyl, tetrazolyl, triazinyl, and thiophenyl (i.e., thienyl).

[0036] The phrase "pharmaceutically acceptable" is used herein to refer to compounds, salts, compositions, dosage forms, etc. that are, within the scope of sound medical judgment, suitable for use in contact with the tissues of humans and / or other animals without undue toxicity, irritation, allergic response, or other problem or complication, commensurate with a reasonable benefit / risk ratio. In some embodiments, "pharmaceutically acceptable" means approved by a federal or state government regulatory agency or listed in the United States Pharmacopeia or other generally recognized pharmacopeia, for use in mammals (e.g., animals), and more particularly humans.

[0037] A "prodrug" is a compound that is metabolized or otherwise chemically converted into an active moiety after administration. A prodrug may be a derivative of an active compound. A prodrug may or may not be active before being converted to the active form in vivo.

[0038] The term "treating" is used herein with reference to methods of treating, for example, a condition or disease, and generally includes administration of a compound or composition to a subject who has not received the compound or composition, to reduce the frequency of or delay the onset of symptoms of the condition or disorder (e.g., cancer, autoimmune disease, inflammatory disorder, gastrointestinal disorder) in the subject. This can include reversing, alleviating, or arresting the symptoms, clinical signs, and underlying pathology of the condition in a manner that improves or stabilizes the subject's condition (e.g., regression of cancer, tissue or cellular senescence, autoimmune or inflammatory disease, etc.).

[0039] The embodiments disclosed herein include all pharmaceutically acceptable compounds of compounds (I) through (IF) that are isotopically labeled by having one or more atoms replaced by an atom having a different atomic mass or mass number. Examples of isotopes that can be incorporated into the disclosed compounds include, respectively: 2 H, 3 H, 11 C. 13 C. 14 C.13 N, 15 N, 15 O. 17 O. 18 O. 31 P, 32 P, 35 S, 18 F, 36 Cl, 123 I, and 125 These radiolabeled compounds may be useful to aid in determining or measuring the efficacy of compounds, for example, by characterizing the site or mode of action, or binding affinity to a pharmacologically important site of action. Certain isotopically labeled compounds of (I)-(IF), for example, compounds incorporating a radioisotope, are useful in drug and / or substrate tissue distribution studies. The radioisotope tritium, i.e., 3 H, and carbon-14, i.e. 14 C are particularly useful for this purpose given their ease of incorporation and ready means of detection.

[0040] Deuterium, i.e., 2 Substitution with heavier isotopes, such as H, may offer certain therapeutic advantages resulting from greater metabolic stability, for example, increased in vivo half-life or reduced dosage requirements, and therefore may be preferable in some circumstances.

[0041] 11 C. 18 F, 15 O, and 13 Substitution with positron-emitting isotopes, such as N, can be useful in positron emission topography (PET) studies to examine substrate receptor occupancy. Isotopically labeled compounds of (I)-(IF) can generally be prepared by conventional techniques known to those skilled in the art, or by processes analogous to those described in the preparations and examples below, using appropriate isotopically labeled reagents in place of previously employed non-labeled reagents.

[0042]

[0013] Embodiments disclosed herein encompass in vivo metabolic products of the disclosed compounds. Such products may result, for example, from oxidation, reduction, hydrolysis, amidation, esterification, etc., of the administered compound, primarily due to enzymatic processes. Thus, the present disclosure includes compounds produced by a process comprising administering a compound of the present disclosure to a mammal for a period of time sufficient to yield its metabolic products. Such products are typically identified by administering a detectable dose of a radiolabeled compound of the present disclosure to an animal, such as a rat, mouse, guinea pig, monkey, or human, allowing a period of time sufficient for metabolism to occur, and isolating the conversion products from urine, blood, or other biological sample.

[0043] "Pharmaceutically acceptable salts" include both acid and base addition salts.

[0044] "Pharmaceutically acceptable acid addition salts" refers to salts that retain the biological effectiveness and properties of the free bases, which are not biologically or otherwise undesirable, and include those salts with inorganic acids such as, but not limited to, hydrochloric acid, hydrobromic acid, sulfuric acid, nitric acid, phosphoric acid, and the like, as well as, but not limited to, acetic acid, 2,2-dichloroacetic acid, adipic acid, alginic acid, ascorbic acid, aspartic acid, benzenesulfonic acid, benzoic acid, 4-acetamidobenzoic acid, camphoric acid, camphor-10-sulfonic acid, capric acid, caproic acid, caprylic acid, carbonic acid, cinnamic acid, citric acid, cyclamic acid, dodecylsulfonic acid, ethane-1,2-disulfonic acid, ethanesulfonic acid, 2-hydroxyethanesulfonic acid, formic acid, fumaric acid, galactaric acid, gentic acid, glucoside, acetic acid, acetic acid, citric ... It is formed from organic acids such as heptonic acid, gluconic acid, glucuronic acid, glutamic acid, glutaric acid, 2-oxoglutaric acid, glycerophosphoric acid, glycolic acid, hippuric acid, isobutyric acid, lactic acid, lactobionic acid, lauric acid, maleic acid, malic acid, malonic acid, mandelic acid, methanesulfonic acid, mucic acid, naphthalene-1,5-disulfonic acid, naphthalene-2-sulfonic acid, 1-hydroxy-2-naphthoic acid, nicotinic acid, oleic acid, orotic acid, oxalic acid, palmitic acid, pamoic acid, propionic acid, pyroglutamic acid, pyruvic acid, salicylic acid, 4-aminosalicylic acid, sebacic acid, stearic acid, succinic acid, tartaric acid, thiocyanic acid, p-toluenesulfonic acid, trifluoroacetic acid (TFA), and undecylenic acid.

[0045] "Pharmaceutically acceptable base addition salts" refer to salts that retain the biological effectiveness and properties of the free acid, and which are not biologically or otherwise undesirable. These salts are prepared from the addition of an inorganic or organic base to the free acid. Salts derived from inorganic bases include, but are not limited to, sodium, potassium, lithium, ammonium, calcium, magnesium, iron, zinc, copper, manganese, aluminum salts, and the like. Preferred inorganic salts are ammonium, sodium, potassium, calcium, and magnesium salts. Salts derived from organic bases include, but are not limited to, salts of primary, secondary, and tertiary amines, substituted amines including naturally occurring substituted amines, cyclic amines, and basic ion exchange resins such as ammonia, isopropylamine, trimethylamine, diethylamine, triethylamine, tripropylamine, diethanolamine, ethanolamine, dianol, 2-dimethylaminoethanol, 2-diethylaminoethanol, dicyclohexylamine, lysine, arginine, histidine, caffeine, procaine, hydrabamine, choline, betaine, benethamine, benzetine, ethylenediamine, glucosamine, methylglucamine, theobromine, triethanolamine, tromethamine, purine, piperazine, piperidine, N-ethylpiperidine, and polyamine resins. Particularly preferred organic bases are isopropylamine, diethylamine, ethanolamine, trimethylamine, dicyclohexylamine, choline, and caffeine.

[0046] A "pharmaceutical composition" refers to a formulation of a compound of the present disclosure and a medium generally accepted in the art for the delivery of biologically active compounds to mammals, e.g., humans. Such a medium includes all pharmaceutically acceptable carriers, diluents, and excipients therefor.

[0047] An "effective amount" or "therapeutically effective amount" refers to the amount of a compound of the present disclosure that, when administered to a mammal, preferably a human, is sufficient to effect treatment in the mammal, preferably a human. The amount of a compound that constitutes a "therapeutically effective amount" varies depending on the compound, the condition and its severity, the method of administration, and the age of the mammal being treated, but can be routinely determined by one of ordinary skill in the art having regard to their own knowledge and this disclosure.

[0048] "Stereoisomer" refers to a compound composed of the same atoms bonded by the same bonds but with different, non-interchangeable three-dimensional structures. The present disclosure contemplates various stereoisomers and mixtures thereof, including "enantiomers," which refer to two stereoisomers whose molecules are non-superimposable mirror images of one another. The present disclosure also contemplates "diastereomers," which refer to non-mirror images of non-identical stereoisomers. Diastereomers occur when two or more stereoisomers of a compound have different configurations at one or more of the equivalent stereocenters and are not mirror images of each other.

[0049] "Tautomer" refers to a proton shift from one atom of a molecule to another atom of the same molecule. The present disclosure includes tautomers of any such compounds.

[0050] A less than complete measure of the disclosure may be claimed for any reason, reserving the right to make exceptions to or exclude any individual member of any such group, including any subrange or combination of subranges within a group, which may be claimed according to ranges or in any similar manner. Further, a less than complete measure of the disclosure may be claimed for any reason, reserving the right to make exceptions to or exclude any individual substituent, analog, compound, ligand, structure, or group thereof, or any member of a claimed group.

[0051] Throughout this disclosure, various patents, patent applications, and publications are referenced. The entire disclosures of these patents, patent applications, and publications are incorporated by reference into this disclosure in order to more fully describe the state of the art as known to those skilled in the art as of the date of this disclosure. This disclosure will control in the event of a conflict between the cited patents, patent applications, and publications and this disclosure.

[0052] II. Compounds The compounds described herein are ionizable lipids with degradable head groups. In one embodiment, a compound having the structure of formula (I), or a stereoisomer, salt, or tautomer thereof, is provided: [ka] During the ceremony, R 1 , R 2 , R 3 , and R 4 are each independently hydrogen, C1 to C 20 Alkyl, C1-C 20 Alkynyl, C1-C 20 Alkenyl, or C1-C 20 is heteroalkyl, and Y 1 and Y 2 are each independently C1-C8 alkyl, C1-C8 alkynyl, C1-C8 alkenyl, or arylC1-C6 alkyl, and X is -OR 5 or -CH2NR 6 R 7 and R 5 is C1-C6 alkyl or C1-C6 heteroalkyl, and R 6 and R 7 are each independently hydrogen, C1-C6 alkyl, or -C(=O)R 8 and R 8 is C1-C6 alkyl, C1-C6 alkynyl, C1-C6 alkenyl, or C1-C6 heteroalkyl.

[0053] In one embodiment, X is -OR5 In other embodiments, X is -CHNR 6 R 7 is.

[0054] In one embodiment, the compound has one of the following structures (IA)-(IB), or a pharmaceutically acceptable salt or stereoisomer thereof: [ka]

[0055] In one embodiment, R 1 , R 2 , R 3 , and R 4 are each independently C1 to C 20 Alkyl, C1-C 20 Alkynyl, or C1-C 20 In another embodiment, R 1 , R 2 , R 3 , and R 4 are each independently C1 to C 20 In another embodiment, R 1 , R 2 , R 3 , and R 4 are each independently C1 to C 20 Alkynyl or C1-C 20 In another embodiment, R 1 , R 2 , R 3 , and R 4 are each independently C1 to C 20 It is alkenyl.

[0056] In one embodiment, R 1 , R 2 , R 3 , and R 4 are each independently C1 to C 18 Alkyl or C1-C 18 In another embodiment, R 1 , R 2 , R3 , and R 4 are each independently C1 to C 18 In another embodiment, R 1 , R 2 , R 3 , and R 4 are each independently C1 to C 18 It is alkenyl.

[0057] In one embodiment, R 1 , R 2 , R 3 , and R 4 are each independently C4 to C 18 Alkyl or C4-C 18 In another embodiment, R 1 , R 2 , R 3 , and R 4 are each independently C4 to C 18 In another embodiment, R 1 , R 2 , R 3 , and R 4 are each independently C4 to C 18 In another embodiment, R 1 , R 2 , R 3 , and R 4 are each independently C 14 ~C 18 It is alkenyl.

[0058] In one embodiment, R 1 , R 2 , R 3 , and R 4 C4~C 18 Alkyl is n-butyl (C4 alkyl), n-pentyl (C5 alkyl), n-hexyl (C6 alkyl), n-heptyl (C7 alkyl), n-octyl (C8 alkyl), n-nonyl (C9 alkyl), n-decanyl (C 10 alkyl), n-undecanyl (C 11 alkyl), n-dodecanyl (C 12 alkyl), n-tridecanyl (C 13alkyl), n-tetradecanyl (C 14 alkyl), n-pentadecanyl (C 15 alkyl), n-hexadecanyl (C 16 alkyl), n-heptadecanyl (C 17 alkyl), or n-octadecanyl (C 18 alkyl).

[0059] In one embodiment, R 1 , R 2 , R 3 , and R 4 C 14 ~C 18 Alkenyl is (6Z,9Z)-hexadeca-6,9-diene or (6Z,9Z)-octadeca-6,9-diene.

[0060] In one embodiment, R 1 , R 2 , R 3 , and R 4 each independently has one of the following structures: [ka]

[0061] In one embodiment, R 1 , R 2 , R 3 , and R 4 each independently has one of the following structures: [ka]

[0062] In one embodiment, R 1 and R 2 One of the two and R 3 and R 4 One of them is [ka] and R 1 and R 2 the other of R 3 and R 4 The other of [ka] is.

[0063] In one embodiment, R 1 and R 2 One of the two and R 3 and R 4 One of them is [ka] and R 1 and R 2 the other of R 3 and R 4 The other of [ka] is.

[0064] In one embodiment, R 1 , R 2 , R 3 , and R 4 are respectively, [ka] is.

[0065] In one embodiment, R 1 , R 2 , R 3 , and R 4 are respectively, [ka] is.

[0066] In one embodiment, R 1 and R 2 One of the two and R 3 and R 4 One of them is [ka] and R 1 and R 2 the other of R 3 and R 4 The other of [ka] is.

[0067] In one embodiment, R 1 and R 2 One of the two and R 3 and R 4 One of them is [ka] and R 1 and R 2 the other of R 3 and R 4 The other of [ka] is.

[0068] In one embodiment, Y 1 and Y 2 are each independently C1-C6 alkyl, C1-C6 alkynyl, C1-C6 alkenyl, or arylC1-C4 alkyl. 1 and Y 2are each independently C1-C6 alkyl. 1 and Y 2 are each independently C1-C6 alkynyl. 1 and Y 2 are each independently C1-C6 alkenyl. 1 and Y 2 are each independently arylC1-C4 alkyl. In another embodiment, Y 1 and Y 2 are each independently phenyl C1-C4 alkyl. In other embodiments, for example, Y 1 and Y 2 The phenyl C1-C4 alkyl is [ka] is.

[0069] In one embodiment, Y 1 and Y 2 are each independently C1-C6 alkyl or C1-C6 alkenyl. 1 and Y 2 are each independently C1-C6 alkyl. 1 and Y 2 are each independently C1-C6 alkenyl.

[0070] In one embodiment, Y 1 and Y 2 are each independently C1-C6 alkyl. In some embodiments, for example, Y 1 and Y 2 are each independently —CH—, —(CH)—, —(CH)—, —(CH)—, —(CH)—, or —(CH)—.

[0071] In one embodiment, the compound has one of the following structures (IC)-(ID), or a pharmaceutically acceptable salt or stereoisomer thereof: [ka] In the formula, n1, n2, n3, and n4 each independently represent an integer of 0 to 19, and m1 and m2 each independently represent an integer of 1 to 8.

[0072] In one embodiment, n1, n2, n3, and n4 are each independently an integer between 0 and 17. In one embodiment, n1, n2, n3, and n4 are each independently an integer between 1 and 17. In one embodiment, n1, n2, n3, and n4 are each independently an integer between 2 and 17. In one embodiment, n1, n2, n3, and n4 are each independently an integer between 3 and 17. In some embodiments, n1, n2, n3, and n4 are each independently an integer between 3 and 17. In some embodiments, n1, n2, n3, and n4 are each independently an integer between 3 and 17. In some embodiments, n1, n2, n3, and n4 are each independently an integer between 4 and 5. In some embodiments, n1, n2, n3, and n4 are each independently an integer between 6 and 7. In some embodiments, n1, n2, n3, and n4 are each independently an integer of 8. In some embodiments, n1, n2, n3, and n4 are each independently an integer of 9. In some embodiments, n1, n2, n3, and n4 are each independently an integer of 10. In some embodiments, n1, n2, n3, and n4 are each independently an integer of 11. In some embodiments, n1, n2, n3, and n4 are each independently an integer of 12. In some embodiments, n1, n2, n3, and n4 are each independently an integer of 13. In some embodiments, n1, n2, n3, and n4 are each independently an integer of 14. In some embodiments, n1, n2, n3, and n4 are each independently an integer of 15. In some embodiments, n1, n2, n3, and n4 are each independently an integer of 16. In some embodiments, n1, n2, n3, and n4 are each independently an integer of 17. In some embodiments, n1, n2, n3, and n4 are each independently an integer of 5, 6, 7, 13, or 17.

[0073] In one embodiment, m1 and m2 are each independently an integer between 1 and 6. In some embodiments, m1 and m2 are each independently an integer between 1 and 6. In some embodiments, m1 and m2 are each independently an integer between 2 and 3. In some embodiments, m1 and m2 are each independently an integer between 3 and 6. In some embodiments, m1 and m2 are each independently an integer between 4 and 6. In some embodiments, m1 and m2 are each independently an integer between 5 and 6. In some embodiments, m1 and m2 are each independently an integer between 1 and 5. In some embodiments, m1 and m2 are each independently an integer from 2 to 5. In some embodiments, m1 and m2 are each independently an integer from 3 to 5. In some embodiments, m1 and m2 are each independently an integer from 4 to 5.

[0074] In one embodiment, R 5 is C1-C4 alkyl or C1-C4 heteroalkyl. 5 is C1-C4 alkyl. In another embodiment, R 5 is C1-C4 heteroalkyl.

[0075] In one embodiment, R 5 is C1-C4 heteroalkyl or C2-C3 heteroalkyl. In other embodiments, R 5 is C1-C4 heteroalkyl. In other embodiments, R 5 is a C2-C3 heteroalkyl.

[0076] In one embodiment, R 5The C2-C3 heteroalkyl in R is further substituted with C1-C6 alkyl, C1-C6 alkynyl, C1-C6 alkenyl, or C1-C6 heteroalkyl. 5 The C2-C3 heteroalkyl in R is further substituted with C1-C6 alkyl. 5 The C2-C3 heteroalkyl in R is further substituted with C1-C6 alkynyl. 5 The C2-C3 heteroalkyl in R is further substituted with C1-C6 alkenyl. 5 The C2-C3 heteroalkyl is further substituted with a C1-C6 heteroalkyl.

[0077] In one embodiment, R 5 is a C1-C4 amine or a C2-C3 amine. In other embodiments, R 5 is a C1-C4 amine. In another embodiment, R 5 is a C2-C3 amine. In another embodiment, R 5 is ethylamine, N-methylethylamine, propylamine, N-methylpropylamine, heptylamine, or N-methylheptylamine.

[0078] In one embodiment, R 5 has the following structure: [ka] It has one of the following.

[0079] In one embodiment, R 6 and R 7 Each of is hydrogen.

[0080] In one embodiment, R 6 and R 7 is hydrogen, and R 6 and R 7 The other of these is C1-C6 alkyl or -C(=O)R 8 In some embodiments, R6 and R 7 is hydrogen, and R 6 and R 7 The other of these is a C1-C6 alkyl.

[0081] In one embodiment, R 6 and R 7 is hydrogen, and R 6 and R 7 The other is -C(=O)R 8 is.

[0082] In one embodiment, the compound has the following structure (IE): or a pharmaceutically acceptable salt or stereoisomer thereof. [ka]

[0083] In one embodiment, R 8 is -CH2NH2 or -CH2NHR 9 where R 9 -CH3, -C z H z+1 , or -(CH2) z OH and Z is an integer from 1 to 6. In another embodiment, R 8 is -CH2NH2.

[0084] In one embodiment, the compound has the following structure (IF): or a pharmaceutically acceptable salt or stereoisomer thereof. [ka] In the formula, R 9 -CH3, -C z H z+1 , or -(CH2) z OH, and Z is an integer of 1 to 6.

[0085] In some embodiments, Z is the integer 1. In some embodiments, Z is the integer 2. In some embodiments, Z is the integer 3. In some embodiments, Z is the integer 4. In some embodiments, Z is the integer 5. In some embodiments, Z is the integer 6. In some embodiments, Z is an integer from 2 to 6. In some embodiments, Z is an integer from 3 to 6. In some embodiments, Z is an integer from 4 to 6. In some embodiments, Z is an integer from 5 to 6. In some embodiments, Z is an integer from 2 to 5. In some embodiments, Z is an integer from 2 to 4. In some embodiments, Z is an integer from 2 to 3. In some embodiments, Z is an integer from 1 to 5. In some embodiments, Z is an integer from 1 to 4. In some embodiments, Z is an integer from 1 to 3. In some embodiments, Z is an integer from 1 to 2.

[0086] In one embodiment, R 9 is -CH3. In some embodiments, R 9 -C z H z+1 , for example, -CH2CH3. In some embodiments, R 9 is -(CH2) z OH, for example, —CH2OH.

[0087] In one embodiment, the compound has one of the following structures shown in Table A below. [Table 1] TIFF2026502350000023.tif242170TIFF2026502350000024.tif200170TIFF2026502350000025.tif254170 TIFF2026502350000026.tif242170TIFF2026502350000027.tif253170TIFF2026502350000028.tif115170

[0088] In one embodiment, the compound shown in Table A is a pharmaceutically acceptable salt. In another embodiment, the pharmaceutically acceptable salt is a trifluoroacetate salt or a hydrochloride salt. In another embodiment, the pharmaceutically acceptable salt is a trifluoroacetate salt. In another embodiment, the pharmaceutically acceptable salt is a hydrochloride salt.

[0089] As can be seen, the ionizable lipids described herein enable the development of new treatments for disease without the need for exotic chemistry or specialized reagents or manufacturing techniques.

[0090] III. Compositions and Uses In some aspects, ionizable lipids and methods for using them are included in compositions such as lipid-nanoparticle (LNP) compositions for the delivery of molecules, particularly therapeutic molecules including therapeutic nucleic acids. In embodiments, compositions containing the ionizable lipids described herein are used to deliver nucleic acids to target cells and / or tissues, such as by transfecting the target cells and / or tissues. More specifically, LNP compositions containing one or more of the ionizable lipids described herein and RNA are administered to subjects for the treatment of pathologies or disorders. Thus, in aspects, the current technology relates to LNP compositions containing the ionizable lipids described herein, as well as methods for using such compositions to deliver nucleic acids to cells and / or tissues, such as by transfecting the cells and / or tissues with the LNPs, ionizable lipid compositions of the present technology.

[0091] In some embodiments, lipid-based delivery systems and methods, such as, but not limited to, lipid nanoparticle (LNP) compositions and methods of using the same, can be used in conjunction with the ionizable lipids described herein. Such ionizable lipid compositions and LNP compositions comprising ionizable lipids, as well as methods of using the same, provide approaches for stabilizing and / or delivering nucleic acids and other therapeutic molecules. In embodiments, ionizable lipid compositions and LNP compositions comprising ionizable lipids, as well as methods of using the same, include design features such as optimal particle size, high encapsulation efficiency, robust manufacturing processes, and / or optimal lipophilicity and surface charge to provide efficient lipid-based delivery systems for nucleic acids and other therapeutic molecules. In some embodiments, the ionizable lipid compositions and LNP compositions comprising ionizable lipids, as well as methods of use, are present in lipid nanoparticle compositions that include additional ingredients and components. In some embodiments, the additional ingredients are present within the lipid nanoparticles themselves.

[0092] In some embodiments, the ionizable lipid compositions and LNP compositions comprising ionizable lipids, and methods provided herein provide for the delivery of a nucleic acid to a target cell or tissue, wherein the nucleic acid may be selected from the group consisting of small interfering RNA (siRNA), asymmetric interfering RNA (aiRNA), microRNA (miRNA), dicer substrate RNA (dsRNA), small hairpin RNA (shRNA), messenger RNA (mRNA), guide RNA (gRNA), plasmid DNA (pDNA), antisense oligodeoxynucleotide (ODN), RNA or DNA vaccines, and mixtures thereof.

[0093] Other embodiments are directed to pharmaceutical compositions. In one embodiment, the pharmaceutical composition comprises any one (or more) of the aforementioned compounds and a pharmaceutically acceptable carrier. In some embodiments, the pharmaceutical composition is formulated for oral administration. In other embodiments, the pharmaceutical composition is formulated for injection. In even more embodiments, the pharmaceutical composition comprises a compound disclosed herein and an additional therapeutic agent (e.g., an anti-cancer agent). Non-limiting examples of such additional therapeutic agents are described herein below.

[0094] Suitable routes of administration include, but are not limited to, oral, intravenous, rectal, aerosol, parenteral, ophthalmic, pulmonary, transmucosal, transdermal, vaginal, ocular, nasal, and topical administration. Additionally, by way of example only, parenteral delivery includes intramuscular, subcutaneous, intravenous, intramedullary injection, as well as intrathecal, direct intraventricular, intraperitoneal, intralymphatic, and intranasal injection.

[0095] In certain embodiments, the compounds described herein are administered in a local rather than systemic manner, for example, via direct injection of the compound into an organ, often in a depot preparation or sustained-release formulation. In specific embodiments, long-acting formulations are administered by implantation (e.g., subcutaneous or intramuscular) or by intramuscular injection. Furthermore, in other embodiments, the compounds are delivered in targeted drug delivery systems, for example, liposomes coated with organ-specific antibodies. In such embodiments, the liposomes are targeted to and selectively taken up by the organ. In still other embodiments, the compounds described herein are provided in the form of a rapid-release formulation, a sustained-release formulation, or an intermediate-release formulation. In still other embodiments, the compounds described herein are administered locally.

[0096] In the treatment methods according to embodiments of the present disclosure, an effective amount of at least one compound of Formula (I)-(IF) is administered to a subject suffering from or diagnosed with such a disease, disorder, or condition. An effective amount or dose may be ascertained by methods such as modeling, dose escalation studies, or clinical trials, taking into account, for example, the mode or route of administration or drug delivery, the pharmacokinetics of the agent, the severity and course of the disease, disorder, or condition, the subject's previous or ongoing therapies, the subject's health status and response to the drug, and the judgment of the treating physician.

[0097] The compounds of the present disclosure are effective over a wide dosage range. For example, in the treatment of adult humans, dosages of about 0.001-0.1 mg, 0.01-0.1 mg, 0.5-5 mg, 0.5-10 mg, 0.01-10 mg, 0.1-10 mg, 10-5000 mg, 100-5000 mg, 1000 mg-4000 mg, or 1000-3000 mg per day are exemplary dosages used in some embodiments. The exact dosage will depend on the route of administration, the form in which the compound is administered, the subject being treated, the subject's weight, and the preference and experience of the attending physician.

[0098] In some embodiments, the compound of the present disclosure is administered in a single dose. In embodiments, the single dose is administered orally. In another embodiment, the single dose is administered topically. However, other routes may be used as needed. In some embodiments, the compound of the present disclosure is administered in multiple doses. In some embodiments, dosing is about once, twice, three times, four times, five times, six times, or more than six times per day. In other embodiments, dosing is about once per month, once every two weeks, once per week, or once every other day. In another embodiment, the compound of the present disclosure and another agent (e.g., an anticancer agent) are administered together about once per day to about six times per day. In another embodiment, administration of the compound of the present disclosure and the agent continues for less than about 7 days. In yet another embodiment, administration continues for more than about 6 days, 10 days, 14 days, 28 days, 2 months, 6 months, or 1 year. In some cases, continuous dosing is implemented and maintained as long as needed.

[0099] The administration of the disclosed compound can continue as long as necessary. In some embodiments, the disclosed compound is administered for more than 1, 2, 3, 4, 5, 6, 7, 14, or 28 days. In some embodiments, the disclosed compound is administered for less than 28, 14, 7, 6, 5, 4, 3, 2, or 1 day. In some embodiments, the disclosed compound is administered chronically on a continuous basis, for example, to treat chronic effects.

[0100] In some embodiments, the compounds of the present disclosure are administered in individual dosage forms. It is known in the art that due to inter-subject variability in the pharmacokinetics of compounds, individualization of dosing regimens is necessary for optimal therapy.

[0101] In some embodiments, the compound described herein is formulated into pharmaceutical composition.In specific embodiments, pharmaceutical composition is formulated by conventional method using one or more physiologically acceptable carriers, including excipients and auxiliary agents that facilitate the processing of the disclosed compound into pharmaceutical preparations.Suitable formulation depends on the selected route of administration. Any pharmaceutically acceptable techniques, carriers, and excipients may be used as suitable for formulating the pharmaceutical compositions described herein: Remington: The Science and Practice of Pharmacy, Nineteenth Ed. (Easton, Pa.: Mack Publishing Company, 1995); Hoover, John E., Remington's Pharmaceutical Sciences, Mack Publishing Co., Easton, Pennsylvania 1975; Liberman, H.A. and Lachman, L., Eds., Pharmaceutical Dosage Forms, Marcel Decker, New York, NY, 1980; and Pharmaceutical Dosage Forms and Drug Delivery Systems, Seventh Ed. (Lippincott Williams & Wilkins 1999).

[0102] Provided herein are compositions (including pharmaceutical compositions) comprising one or more compounds of Formulae (I)-(IF) and a pharmaceutically acceptable carrier. Also provided herein are pharmaceutical compositions comprising one or more compounds selected from the compounds of Formulae (I)-(IF) and pharmaceutically acceptable diluent(s), excipient(s), and carrier(s). In certain embodiments, the described compounds are administered as pharmaceutical compositions, similar to combination therapies, in which one or more compounds selected from the compounds of Formulae (I)-(IF) are mixed with other active ingredients. All combinations of active agents described in the Combination Therapy section below and throughout this disclosure are encompassed herein. In specific embodiments, the pharmaceutical composition comprises one or more compounds of Formulae (I)-(IF).

[0103] As used herein, a pharmaceutical composition refers to a mixture of one or more compounds selected from the compounds of Formulae (I)-(IF) with other chemical components, such as carriers, stabilizers, diluents, dispersants, suspending agents, thickeners, and / or excipients. In certain embodiments, the pharmaceutical composition facilitates administration of the compound to an organism. In some embodiments, a therapeutically effective amount of one or more compounds selected from the compounds of Formulae (I)-(IF) provided herein is administered in the pharmaceutical composition to a mammal having a disease, disorder, or medical condition to be treated. In specific embodiments, the mammal is a human. In certain embodiments, the therapeutically effective amount varies depending on the severity of the disease, the age and relative health of the subject, the potency of the compound used, and other factors. The compounds described herein are used alone or in combination with one or more therapeutic agents as components of a mixture.

[0104] In one embodiment, one or more compounds selected from the compounds of Formulae (I)-(IF) are formulated in an aqueous solution. In a specific embodiment, the aqueous solution is selected from a physiologically compatible buffer, such as, by way of example only, Hank's solution, Ringer's solution, or physiological saline buffer. In other embodiments, one or more compounds selected from the compounds of Formulae (I)-(IF) are formulated for transmucosal administration. In a specific embodiment, the transmucosal formulation includes a penetrant appropriate for the barrier to be permeated. In still other embodiments, where the compounds described herein are formulated for other parenteral injections, suitable formulations include aqueous or non-aqueous solutions. In a specific embodiment, such solutions include physiologically compatible buffers and / or excipients.

[0105] In another embodiment, the compound described herein is formulated for oral administration.The compound described herein is formulated by combining active compound with, for example, pharmaceutically acceptable carrier or excipient.In various embodiments, the compound described herein is formulated into oral dosage forms, including, for example, tablets, powders, pills, dragees, capsules, liquids, gels, syrups, elixirs, slurries, suspensions, etc.

[0106] In certain embodiments, oral medicaments are prepared by mixing one or more solid excipients with one or more of the compounds described herein, optionally grinding the resulting mixture, and then processing the resulting mixture with suitable additives, if desired, to obtain tablets or dragee cores.Suitable excipients include fillers such as sugars, including lactose, sucrose, mannitol, or sorbitol, corn starch, wheat starch, rice starch, potato starch, gelatin, tragacanth gum, methylcellulose, microcrystalline cellulose, hydroxypropylmethylcellulose, sodium carboxymethylcellulose, or other cellulose preparations, such as polyvinylpyrrolidone (PVP or povidone) or calcium phosphate.In certain embodiments, disintegrants are optionally added.Disintegrants include, by way of example only, cross-linked croscarmellose sodium, polyvinylpyrrolidone, agar, or alginic acid or its salt, such as sodium alginate.

[0107] In one embodiment, oral dosage forms such as pills, capsules, or tablets include one or more suitable layers or coatings. In a specific embodiment, a concentrated sugar solution is used to coat the dosage form. The sugar solution optionally contains additional ingredients such as, for example, gum arabic, talc, polyvinylpyrrolidone, carbopol gel, polyethylene glycol, and / or titanium dioxide, a lacquer solution, and a suitable organic solvent or solvent mixture. Dyes and / or pigments are also optionally added to the coating for identification purposes. Additionally, dyes and / or pigments are optionally used to distinguish different combinations of active compound doses.

[0108] In certain embodiments, a therapeutically effective amount of at least one of the compounds described herein is formulated into other oral dosage forms. Oral dosage forms include push-fit capsules made of gelatin and soft sealed capsules made of gelatin and a plasticizer such as glycerol or sorbitol. In specific embodiments, the push-fit capsules contain the active ingredient mixed with one or more fillers. Fillers include, by way of example only, binders such as lactose, starch, and / or lubricants such as talc or magnesium stearate, and optionally, stabilizers. In other embodiments, soft capsules contain one or more active compounds dissolved or suspended in a suitable liquid. Suitable liquids include, by way of example only, one or more fatty oils, liquid paraffin, or liquid polyethylene glycol. In addition, stabilizers are optionally added.

[0109] In still other embodiments, the compounds described herein are formulated for parenteral injection, including formulations suitable for bolus injection or continuous infusion. In specific embodiments, the injectable formulations are presented in unit dosage form (e.g., in ampoules) or in multi-dose containers. A preservative is optionally added to the injectable formulation. In still other embodiments, the pharmaceutical composition is formulated in a form suitable for parenteral injection as a sterile suspension, solution, or emulsion in an oily or aqueous vehicle. Parenteral injection formulations optionally contain formulating agents such as suspending agents, stabilizers, and / or dispersing agents. In specific embodiments, pharmaceutical formulations for parenteral administration comprise an aqueous solution of the active compound in water-soluble form. In additional embodiments, a suspension of one or more compounds selected from the compounds of Formulae (I)-(IF) is prepared as a suitable oily injection suspension. Suitable lipophilic solvents or vehicles for use in the pharmaceutical compositions described herein include, by way of example only, fatty oils such as sesame oil, or synthetic fatty acid esters such as ethyl oleate or triglycerides, or liposomes. In certain specific embodiments, aqueous injection suspensions contain substances that increase the viscosity of the suspension, such as sodium carboxymethylcellulose, sorbitol, or dextran. Optionally, the suspension contains suitable stabilizers or agents that increase the solubility of the compounds, allowing for the preparation of highly concentrated solutions. Alternatively, in other embodiments, the active ingredient is in powder form for constitution with a suitable vehicle, for example, sterile, pyrogen-free water, before use.

[0110] Pharmaceutical compositions comprise at least one pharmaceutically acceptable carrier, diluent, or excipient and one or more compounds selected from the compounds of Formulae (I) through (IF) as an active ingredient. The active ingredient may be in the form of a free acid or free base, or in the form of a pharmaceutically acceptable salt. Additionally, the methods and pharmaceutical compositions described herein include the use of N-oxides, crystalline forms (also known as polymorphs), and active metabolites of these compounds having the same type of activity. All tautomers of the compounds described herein are included within the scope of the compounds presented herein. Additionally, the compounds described herein encompass unsolvated and solvated forms with pharmaceutically acceptable solvents, such as water, ethanol, and the like. Solvated forms of the compounds presented herein are also considered to be disclosed herein. Additionally, pharmaceutical compositions optionally contain other medicinal or pharmaceutical agents, carriers, adjuvants such as preservatives, stabilizers, wetting agents, or emulsifiers, solubility enhancers, salts for regulating osmotic pressure, buffers, and / or other therapeutically valuable substances.

[0111] Methods for preparing compositions containing the compounds described herein include formulating the compound(s) with one or more inert pharmaceutically acceptable excipients or carriers to form solid, semi-solid, or liquid compositions. Solid compositions include, but are not limited to, powders, tablets, dispersible granules, capsules, cachets, and suppositories. Liquid compositions include solutions in which the compounds are dissolved, emulsions containing the compounds, or solutions containing liposomes, micelles, or nanoparticles containing the compounds disclosed herein. Semi-solid compositions include, but are not limited to, gels, ointments, suspensions, and creams. The pharmaceutical compositions described herein may be in the form of liquid solutions or suspensions, solid forms suitable for dissolution or suspension in liquid before use, or as emulsions. These compositions also optionally contain minor amounts of non-toxic auxiliary substances, such as wetting or emulsifying agents, pH buffering agents, and the like.

[0112] In some embodiments, pharmaceutical compositions comprising one or more compounds selected from the compounds of Formulae (I)-(IF) are illustratively in the form of a liquid, with the drug being present in solution, suspension, or both. Typically, when the composition is administered as a suspension, a first portion of the drug is present in solution, and a second portion of the drug is present in particulate form in suspension in a liquid matrix. In some embodiments, the liquid composition comprises a gel formulation. In other embodiments, the liquid composition is aqueous.

[0113] In certain embodiments, aqueous suspensions contain one or more polymers as suspending agents.Polymers include water-soluble polymers such as cellulose-based polymers, for example, hydroxypropylmethylcellulose, and water-insoluble polymers such as cross-linked carboxyl-containing polymers.Certain pharmaceutical compositions described herein include mucoadhesive polymers selected from, for example, carboxymethylcellulose, carbomer (acrylic acid polymer), poly(methyl methacrylate), polyacrylamide, polycarbophil, acrylic acid / butyl acrylate copolymer, sodium alginate, and dextran.

[0114] The pharmaceutical composition also optionally includes a solubilizing agent to aid in the solubility of one or more compounds selected from the compounds of Formulae (I)-(IF). The term "solubilizing agent" generally includes agents that result in the formation of a micellar or true solution of the drug. Certain acceptable nonionic surfactants, such as polysorbate 80, are useful as solubilizing agents, as are ophthalmically acceptable glycols, polyglycols, such as polyethylene glycol 400, and glycol ethers.

[0115] In addition, the pharmaceutical compositions optionally contain one or more pH adjusting or buffering agents, including acids such as acetic acid, boric acid, citric acid, lactic acid, phosphoric acid, and hydrochloric acid, bases such as sodium hydroxide, sodium phosphate, sodium borate, sodium citrate, sodium acetate, sodium lactate, and trishydroxymethylaminomethane, and buffers such as citrate / dextrose, sodium bicarbonate, and ammonium chloride, etc. Such acids, bases, and buffers are included in amounts necessary to maintain the pH of the composition within an acceptable range.

[0116] The composition also optionally contains one or more salts in an amount necessary to bring the osmolality of the composition into an acceptable range. Such salts include those having sodium, potassium, or ammonium cations and chloride, citrate, ascorbate, borate, phosphate, bicarbonate, sulfate, thiosulfate, or bisulfite anions; suitable salts include sodium chloride, potassium chloride, sodium thiosulfate, sodium bisulfite, and ammonium sulfate.

[0117] Other pharmaceutical compositions optionally contain one or more preservatives to inhibit microbial activity. Suitable preservatives include mercury-containing substances such as merfen and thiomersal, stabilized chlorine dioxide, and quaternary ammonium compounds such as benzalkonium chloride, cetyltrimethylammonium bromide, and cetylpyridinium chloride.

[0118] The composition may contain one or more surfactants to enhance physical stability or for other purposes. Suitable nonionic surfactants include polyoxyethylene fatty acid glycerides and vegetable oils, such as polyoxyethylene (60) hydrogenated castor oil, and polyoxyethylene alkyl ethers and alkylphenyl ethers, such as Octoxynol 10 and Octoxynol 40.

[0119] The compositions may optionally include one or more antioxidants to enhance chemical stability. Suitable antioxidants include, by way of example only, ascorbic acid and sodium metabisulfite.

[0120] In certain embodiments, the aqueous suspension compositions are packaged in single-dose non-reclosable containers. Alternatively, multi-dose reclosable containers are used, in which case it is typical to include a preservative in the composition.

[0121] In alternative embodiments, other delivery systems for hydrophobic pharmaceutical compounds are employed. Liposomes and emulsions are examples of delivery vehicles or carriers useful herein. In certain embodiments, organic solvents such as N-methylpyrrolidone are also employed. In additional embodiments, the compounds described herein are delivered using sustained-release systems, such as semipermeable matrices of solid hydrophobic polymers containing therapeutic agents. Various sustained-release materials are useful herein. In some embodiments, sustained-release capsules release the compound for several weeks up to over 100 days. Depending on the chemical nature and biological stability of the therapeutic reagent, additional strategies for protein stabilization are employed.

[0122] In certain embodiments, the compositions or formulations described herein comprise one or more antioxidants, metal chelators, thiol-containing compounds, and / or other general stabilizing agents. Examples of such stabilizers include, but are not limited to, (a) about 0.5% to about 2% w / v glycerol, (b) about 0.1% to about 1% w / v methionine, (c) about 0.1% to about 2% w / v monothioglycerol, (d) about 1 mM to about 10 mM EDTA, (e) about 0.01% to about 2% w / v ascorbic acid, (f) 0.003% to about 0.02% w / v polysorbate 80, (g) 0.001% to about 0.05% w / v polysorbate 20, (h) arginine, (i) heparin, (j) dextran sulfate, (k) cyclodextrin, (l) pentosan polysulfate and other heparinoids, (m) divalent cations such as magnesium and zinc, or (n) combinations thereof.

[0123] In some embodiments, the concentration of one or more compounds selected from the compounds of Formulae (I)-(IF) provided in the pharmaceutical composition is 95%, 90%, 85%, 80%, 75%, 70%, 65%, 60%, 55%, 50%, 45%, 40%, 35%, 30%, 25%, 20%, 19.75%, 19.50%, 19.25%, 19%, 18.75%, 18.50%, 18.25%, 18%, 17.75%, 17.50%, 17.25%, 17%, 16.75%, 17.50%, 17.25%, 17%, 16.75%, 17.50%, 17.25%, 17%, 17.50 ... %, 16.50%, 16.25%, 16%, 15.75%, 15.50%, 15.25%, 15%, 14.75%, 14.50%, 14.25%, 14%, 13.75%, 13.50%, 13.25%, 13%, 12.75%, 12.50%, 12.25%, 12%, 11.75%, 11.50%, 11.25%, 11%, 10.75%, 10.50%, 10.25%, 10%, 9.75%, 9.50%, 9.25%, 9%, 8.75%, 8.50% ,8.25%,8%,7.75%,7.50%,7.25%,7%,6.75%,6.50%,6.25%,6%,5.75%,5.50%,5.25%,5%,4.75%,4.50%,4.25%,4%,3.75%,3.50%,3.25%,3%,2.75%,2.50%,2.25%,2%,1.75%,1.50%,1.25%,1%,0.5%,0.4%,0.3%,0.2%,0.1%,0.09%,0.08%,0.07%,0. 0.06%, 0.05%, 0.04%, 0.03%, 0.02%, 0.01%, 0.009%, 0.008%, 0.007%, 0.006%, 0.005%, 0.004%, 0.003%, 0.002%, 0.001%, 0.0009%, 0.0008%, 0.0007%, 0.0006%, 0.0005%, 0.0004%, 0.0003%, 0.0002%, or greater than 0.0001% w / w, w / v, v / v, or mole / mole (mole fraction).In another embodiment, the amount of the compound selected from the compounds of Formulae (I)-(IF) in the pharmaceutical composition is between about any two of the values ​​listed in the preceding sentence, e.g., about 2-70 w / w, w / v, v / v, or mol / mol, 3.5-80 w / w, w / v, v / v, or mol / mol, 1-30 w / w, w / v, v / v, or mol / mol, 2-90 w / w, w / v, v / v, or mol / mol, 1-70 w / w, v / v, or mol / mol, 1-80 w / w, w / v, v / v, or mol / mol, 1-90 w / w, w / v, v / v, or mol / mol, etc.

[0124] In some embodiments, the concentration of one or more compounds selected from the compounds of Formulae (I) to (IF) provided in the pharmaceutical compositions of the present disclosure is from about 0.0001% to about 50%, from about 0.001% to about 40%, from about 0.01% to about 30%, from about 0.02% to about 29%, from about 0.03% to about 28%, from about 0.04% to about 27%, from about 0.05% to about 26%, from about 0.06% to about 25%, from about 0.07% to about 10%, from about 0.08% to about 12%, from about 0.09% to about 13%, from about 0.10% to about 14%, from about 0.11% to about 15%, from about 0.12% to about 16%, from about 0.13% to about 17%, from about 0.14% to about 18%, from about 0.15% to about 19%, from about 0.16% to about 20%, from about 0.17% to about 22%, from about 0.18% to about 23%, from about 0.19% to about 24%, from about 0.19% to about 25%, from about 0.19% to about 26%, from about 0.19% to about 26%, from about 0.19% to about 27%, from about 0.19% to about 28%, from about 0.19% to about 29%, from about 0.19% to about 28%, from about 0.19% to about 29%, from about 0.19% to about 25%, from about 0.19% to about 26%, from about 0.19% to about 25%, from about 0.19% to about 26%, from about 0.19% to about % to about 24%, about 0.08% to about 23%, about 0.09% to about 22%, about 0.1% to about 21%, about 0.2% to about 20%, about 0.3% to about 19%, about 0.4% to about 18%, about 0.5% to about 17%, about 0.6% to about 16%, about 0.7% to about 15%, about 0.8% to about 14%, about 0.9% to about 12%, about 1% to about 10% w / w, w / v, v / v, or mol / mol.

[0125] In some embodiments, the amount of one or more compounds selected from compounds of Formulae (I)-(IF) provided in the pharmaceutical composition of the present disclosure is 10 g, 9.5 g, 9.0 g, 8.5 g, 8.0 g, 7.5 g, 7.0 g, 6.5 g, 6.0 g, 5.5 g, 5.0 g, 4.5 g, 4.0 g, 3.5 g, 3.0 g, 2.5 g, 2.0 g, 1.5 g, 1.0 g, 0.95 g, 0.9 g, 0.85 g, 0.8 g, 0.75 g, 0.7 g, 0.65 g, 0.6 g, 0.55 g, 0.5 g, 0.45 g, 0.4 g, 0.35 g, 0.45 g, 0.55 g, 0.55 g, 0.45 g, 0.45 g, 0.55 g, 0.55 g, 0.55 g, 0.55 g, 0.45 g, 0.45 g, 0.45 g, 0.45 g, 0.45 g, 0.45 g, 0.45 g, 0.45 g, 0.5 ... g, 0.3 g, 0.25 g, 0.2 g, 0.15 g, 0.1 g, 0.09 g, 0.08 g, 0.07 g, 0.06 g, 0.05 g, 0.04 g, 0.03 g, 0.02 g, 0.01 g, 0.009 g, 0.008 g, 0.007 g, 0.006 g, 0.005 g, 0.004 g, 0.003 g, 0.002 g, 0.001 g, 0.0009 g, 0.0008 g, 0.0007 g, 0.0006 g, 0.0005 g, 0.0004 g, 0.0003 g, 0.0002 g, or 0.0001 g.

[0126] In some embodiments, the amount of one or more compounds selected from the compounds of Formulae (I)-(IF) provided in the pharmaceutical compositions of the present disclosure is within the range of 0.0001-10 g, 0.0005-9 g, 0.001-8 g, 0.005-7 g, 0.01-6 g, 0.05-5 g, 0.1-4 g, 0.5-4 g, or 1-3 g.

[0127] Packaging materials for use in packaging the pharmaceutical compositions described herein include, for example, those found in U.S. Patent Nos. 5,323,907, 5,052,558, and 5,033,252. Examples of pharmaceutical packaging materials include, but are not limited to, blister packs, bottles, tubes, inhalers, pumps, bags, vials, containers, syringes, bottles, and any packaging material suitable for the selected formulation and intended mode of administration and treatment. For example, a container(s) contains one or more compounds described herein, optionally in a composition or in combination with another drug disclosed herein. The container(s) optionally has a sterile access port (e.g., the container is an intravenous solution bag or a vial with a stopper that can be punctured by a hypodermic needle). Such kits optionally contain the compound together with an identifying description or label or instructions for its use in the methods described herein.

[0128] For example, a kit typically includes one or more additional containers, each containing one or more of various materials (such as reagents and / or devices, optionally in concentrated form) that are desirable from a commercial and user perspective for use of the compounds described herein. Non-limiting examples of such materials include, but are not limited to, buffers, diluents, filters, needles, syringes, carriers, packaging, container, vial, and / or tube labels describing the contents and / or instructions for use, and package inserts with instructions. A set of instructions for use is also typically included. The label is optionally on or associated with the container. For example, a label is on the container when letters, numbers, or other characters forming the label are attached, molded, or etched into the container itself, and a label is associated with the container when present in a receptacle or carrier that also holds the container, for example, as a package insert. In addition, the label is used to indicate that the contents are to be used for a particular therapeutic application. In addition, the label indicates instructions for using the contents, such as in the methods described herein. In certain embodiments, the pharmaceutical compositions are presented in a pack or dispenser device containing one or more unit dosage forms containing a compound provided herein. For example, the pack contains metal or plastic foil, such as a blister pack. Alternatively, the pack or dispenser device is accompanied by instructions for administration. Alternatively, the pack or dispenser carries a notice associated with the container in a form prescribed by a government agency regulating the manufacture, use, or sale of pharmaceuticals, which notice reflects the agency's approval of the drug form for human or veterinary administration. Such notice may be, for example, labeling approved for prescription drugs by the U.S. Food and Drug Administration or an approved product insert. In some embodiments, a composition containing a compound provided herein formulated with a compatible pharmaceutical carrier is prepared, placed in an appropriate container, and labeled for the treatment of an indicated condition.

[0129] As mentioned above, the compounds and compositions of the present disclosure find utility in a wide range of diseases and conditions mediated by protein kinase, including kinase-mediated diseases and conditions.Such diseases include, but are not limited to, lung cancer, NSCLC (non-small cell lung cancer), oat cell carcinoma, bone cancer, pancreatic cancer, skin cancer, dermatofibrosarcoma protuberans, head and neck cancer, skin or intraocular melanoma, uterine cancer, ovarian cancer, colorectal cancer, anal region cancer, stomach cancer, colon cancer, breast cancer, gynecological tumors (for example, uterine sarcoma, fallopian tube cancer, endometrial cancer, cervical cancer, vaginal cancer or vulvar cancer), Hodgkin's disease, hepatocellular carcinoma, esophageal cancer, small intestine cancer, endocrine system cancer (for example, thyroid cancer, pancreatic cancer, parathyroid cancer or adrenal cancer), soft tissue sarcoma, urethral cancer, penile cancer, prostate cancer (particularly hormone-refractory), These may include cancers such as chronic or acute leukemia, childhood solid tumors, hypereosinophilia, lymphocytic lymphoma, bladder cancer, kidney or ureter cancer (e.g., renal cell carcinoma, renal pelvic cancer), childhood malignancies, tumors of the central nervous system (e.g., primary CNS lymphoma, spinal axis tumor, medulloblastoma, brainstem glioma, or pituitary adenoma), Barrett's esophagus (premalignant syndrome), neoplastic skin diseases, psoriasis, mycosis, and benign prostatic hyperplasia, diabetes-related diseases such as diabetic retinopathy, retinal ischemia, and retinal neovascularization, cardiovascular diseases such as liver cirrhosis, angiogenesis, atherosclerosis, immune diseases such as autoimmune diseases, and renal diseases.

[0130] In some embodiments, the pharmaceutical composition comprises a compound described above and a pharmaceutically acceptable carrier, including, for example, any adjuvant, carrier, excipient, glidant, sweetener, diluent, preservative, dye / colorant, flavor enhancer, surfactant, wetting agent, dispersing agent, suspending agent, stabilizer, isotonic agent, solvent, or emulsifier, approved by the U.S. Food and Drug Administration as acceptable for use in humans or veterinary animals.

[0131] In some embodiments, the method of treating a disease or disorder comprises administering an effective amount of a compound or pharmaceutical composition described herein to a subject in need thereof. In some embodiments, the disease or disorder is a kinase-expressing cancer. In some specific embodiments, the cancer is bladder cancer. In some other specific embodiments, the cancer is prostate cancer. In some other specific embodiments, the cancer is a hematological malignancy such as acute myeloid leukemia. In some other specific embodiments, the disease or disorder is an autoimmune disease or an inflammatory disease.

[0132] In one embodiment, the pharmaceutical composition comprises a compound of Formula (I)-(IF) and a therapeutic agent comprising a nucleic acid.

[0133] In one embodiment, the pharmaceutical composition comprising a compound of Formula (I)-(IF) and a therapeutic agent further comprises a helper lipid, a stabilizing lipid, or a structural lipid.

[0134] In one embodiment, the helper lipid is 1,2-dilinoleoyl-sn-glycero-3-phosphocholine (DLPC), 1,2-dimyristoyl-sn-glycero-phosphocholine (DMPC), 1,2-dioleoyl-sn-glycero-3-phosphocholine (DOPC), 1,2-dipalmitoyl-sn-glycero-3-phosphocholine (DPPC), 1,2-distearoyl-sn-glycero-3-phosphocholine (DSPC), 1,2-dioleoyl-sn-glycero-3-phosphocholine (DOPC), 1,2-dioleoyl-sn-glycero-3-phosphocholine (DPPC), 1,2-distearoyl-sn-glycero-3-phosphocholine (DSPC), 1,2-dioleoyl-sn-glycero-3-phosphocholine (DOPC), 1,2-dioleoyl-sn-glycero-3-phosphocholine (DOPC), 1,2-dioleoyl-sn-glycero-3-phosphocholine (DPPC), 1,2-dioleoyl-sn-glycero-3-phosphocholine (DSPC), 1,2-dioleoyl-sn-glycero-3-phosphocholine (DO ... 1-O-undecanoyl-sn-glycero-phosphocholine (DUPC), 1-palmitoyl-2-oleoyl-sn-glycero-3-phosphocholine (POPC), 1,2-di-O-octadecenyl-sn-glycero-3-phosphocholine (18:0 diether PC), l-oleoyl-2-cholesterylhemisuccinoyl-sn-glycero-3-phosphocholine (OChemsPC), 1-hexadecyl-sn-glycero-3-phosphocholine (C16 Lyso PC), 1,2-dilinolenoyl-sn-glycero-3-phosphocholine, 1,2-diarachidonoyl-sn-glycero-3-phosphocholine, 1,2-didocosahexaenoyl-sn-glycero-3-phosphocholine, 1,2-dioleoyl-sn-glycero-3-phosphoethanolamine (DOPE), 1,2-diphytanoyl-sn-glycero-3-phosphoethanolamine (ME 16.0 PE), 1,2-distearoyl-sn-glycero-3-phosphoethanolamine, 1,2-dilinoleoyl-sn-glycero-3-phosphoethanolamine, 1,2-dilinolenoyl-sn-glycero-3-phosphoethanolamine, 1,2-diarachidonoyl-sn-glycero-3-phosphoethanolamine, 1,2-didocosahexaenoyl-sn-glycero-3-phosphoethanolamine, 1,2-dioleoyl-sn-glycero-3-phospho-rac-(1-glycerol) sodium salt (DOPG), and mixtures thereof.

[0135] In one embodiment, the stabilizing lipid is 1-(monomethoxy-polyethylene glycol)-2,3-dimyristoylglycerol (PEG-DMG) with an average PEG molecular weight of 2000.

[0136] In one embodiment, the structural lipid is selected from the group consisting of cholesterol, cholesterol derivatives, fecosterol, sitosterol, ergosterol, campesterol, stigmasterol, brassicasterol, tomatidine, ursolic acid, alpha tocopherol, and mixtures thereof.

[0137] In one embodiment, the nucleic acid is selected from small interfering RNA (siRNA), asymmetric interfering RNA (aiRNA), microRNA (miRNA), dicer substrate RNA (dsRNA), small hairpin RNA (shRNA), or messenger RNA (mRNA).

[0138] In one embodiment, the nucleic acid is mRNA.

[0139] In one embodiment, a pharmaceutical composition comprising a compound of Formula (I)-(IF) and a second therapeutic agent.

[0140] In one embodiment, a method of administering a therapeutic agent comprising a nucleic acid to a patient in need thereof, comprising administering a pharmaceutical composition to the patient.

[0141] In one embodiment, the patient is an animal, hi another embodiment, the patient is a human.

[0142] In one embodiment, a lipid nanoparticle comprising a compound of Formula (I)-(IF) and a therapeutic agent comprising a nucleic acid.

[0143] In one embodiment, the therapeutic agent in the lipid nanoparticle is mRNA.

[0144] The lipid component of the lipid-nanoparticle composition can comprise one or more structural lipids.The structural lipid can be selected from the group consisting of, but not limited to, cholesterol, fecosterol, sitosterol, ergosterol, campesterol, stigmasterol, brassicasterol, tomatidine, ursolic acid, alpha-tocopherol, and mixtures thereof.In some embodiments, the structural lipid is cholesterol.In some embodiments, the structural lipid includes cholesterol and corticosteroids (such as prednisolone, dexamethasone, prednisone, and hydrocortisone), or a combination thereof.

[0145] Structural lipids can also include natural and synthetic cholesterol derivatives. Some examples of natural cholesterol derivatives include, but are not limited to, 7β-OHC, 22(R)-hydroxycholesterol (22R-OHC), 24(S)-hydroxycholesterol (24(S)-OHC). Synthetic cholesterol derivatives include 22(R)-hydroxy-Δ9-cholestanol (22R-ISO-OHC), 23-(4-methylfuran-2,5-dione)-3α-hydroxy-24-nor-5β-cholane (LITHO 1a), 23-(4-methylfuran-2,5-dione)-3α,7α-dihydroxy-24-nor-5β-cholane (CHENO 1b), 23-(4-methyl-1H-pyrrole-2,5-dione)-3α-hydroxy-24-nor-5β-cholane (LITOMAL 7a), and 23-(4-methyl-1H-pyrrole-2,5-dione)-3α,7α,12α-trihydroxy-24-nor-5β-cholane (COLMAL 7f), and the ethanolamide derivatives of lithocholic acid and chenodeoxycholic acid (LITOMET, CHENOMET) (146, 147). The systematic name of LITOMET is (23-((2-hydroxyethyl)-4-methyl-1H-pyrrole-2,5-dione)-3α-hydroxy-24-nor-5β-cholane), and the systematic name of CHENOMET is (23-((2-hydroxyethyl)-4-methyl-1H-pyrrole-2,5-dione)-3α,7α-dihydroxy-24-nor-5β-cholane).

[0146] The lipid-nanoparticle compositions of the present disclosure include nucleic acids such as, but not limited to, small interfering RNA (siRNA), asymmetric interfering RNA (aiRNA), microRNA (miRNA), miRNA inhibitors (antagomir / antimir), dicer substrate RNA (dsRNA), small hairpin RNA (shRNA), messenger RNA (mRNA), polyvalent RNA, and mixtures thereof.

[0147] The lipid-nanoparticle compositions of the present disclosure can be prepared by mixing processes such as, but not limited to, microfluidics and T-junction mixing of two fluid streams, one of which contains nucleic acid and the other of which has a lipid component, which induces nanoprecipitation and particle formation.

[0148] The lipid-nanoparticle compositions of the present disclosure can be characterized using a Zetasizer to determine particle size, polydispersity index (PDI), and zeta potential. In some embodiments, the zeta potential of the lipid-nanoparticle composition can be about -10 mV to about +20 mV, about -10 mV to about +15 mV, about -10 mV to about +10 mV, about -10 mV to about +5 mV, about -10 mV to about 0 mV, about -10 mV to about -5 mV, about -5 mV to about +20 mV, about -5 mV to about +15 mV, about -5 mV to about +10 mV, about -5 mV to about +5 mV, about -5 mV to about 0 mV, about 0 mV to about +20 mV, about 0 mV to about +15 mV, about 0 mV to about +10 mV, about 0 mV to about +5 mV, about +5 mV to about +20 mV, about +5 mV to about +15 mV, or about +5 mV to about +10 mV.

[0149] UV-visible spectroscopy can be used to determine the concentration of nucleic acids in the nanoparticle composition.

[0150] The lipid-nanoparticle compositions of the present disclosure can induce expression of a desired protein both in vitro and in vivo by contacting cells with lipid nanoparticles comprising one or more of the ionizable lipids described herein, where the lipid nanoparticles are encapsulated with or associated with a nucleic acid (e.g., mRNA) that will be expressed to produce the desired protein.

[0151] The lipid-nanoparticle compositions of the present disclosure can reduce target gene and protein expression both in vitro and in vivo by contacting cells with lipid nanoparticles comprising one or more of the ionizable lipids described herein, where the lipid nanoparticles are encapsulated with or associated with a nucleic acid (e.g., siRNA) that reduces target gene expression.

[0152] The lipid-nanoparticle compositions of the present disclosure can downregulate or silence expression of target genes and proteins both in vitro and in vivo by contacting cells with lipid nanoparticles comprising one or more of the ionizable lipids described herein, wherein the lipid nanoparticles are encapsulated with or associated with a nucleic acid that downregulates or silences target gene expression.

[0153] IV. Method of Use The compounds of Formulas (I)-(IF) are stable at low pH and can encapsulate mRNA. At higher pH, the compounds of Formulas (I)-(IF) undergo rapid degradation. In one embodiment, the pH at which the compounds of Formulas (I)-(IF) begin to degrade is physiological conditions. In certain embodiments, the pH at which the compounds of Formulas (I)-(IF) begin to degrade is 7.4 or higher. The degradation process promotes the release and delivery of mRNA. The degradation process of the compounds of Formulas (I), (IB), (ID), and (IE) is illustrated below. [ka]

[0154] The decomposition process of compounds of formula (I), (IC), and (IF) is illustrated below. [ka]

[0155] The following reaction schemes illustrate methods for making lipids of formulas (I), (II), (III), (IV), (V), (VI), and (VII).

[0156] The synthesis of compounds I and II is described in the following scheme. [ka]

[0157] Step 1: In a dry 100 mL round-bottom flask, 60% NaH (225 mg, 5.6 mmol, 1.25 equiv.) dispersed in oil was added under a nitrogen atmosphere. Anhydrous THF (18 mL) and anhydrous DMF (5 mL) were added slowly, sequentially, via syringe. After 10 min, (S)-(+)-2,2-dimethyl-1,3-dioxolane-4-methanol 1 (0.5 mL, 4.5 mmol, 1 equiv.) was added slowly via syringe, and the resulting reaction mixture was stirred at room temperature for 1 h. 4-Methoxybenzyl chloride (0.67 mL, 5 mmol, 1.1 equiv.) and tetrabutylammonium iodide (59 mg, 0.16 mmol, 0.036 equiv.) were added, and the reaction mixture was stirred for an additional 12 h. MeOH (2 mL) was added. The reaction mixture was then washed with saturated aqueous ammonium chloride (25 mL), water (25 mL), and brine (25 mL). The aqueous layers were combined and back-extracted with DCM (2 x 50 mL). The organic layers were combined, dried over anhydrous sodium sulfate, filtered, and concentrated in vacuo to provide the crude product, which was purified by silica gel flash chromatography using 0-10% ethyl acetate in hexanes to provide Intermediate 2 (980 mg, 96% yield).

[0158] Step 2: To a solution of intermediate 2 (1 g, 4 mmol, 1 equiv.) in methanol (5 mL), p-toluenesulfonic acid (38 mg, 0.2 mmol, 0.05 equiv.) was added, and the resulting reaction mixture was stirred at room temperature for 72 h. Triethylamine (0.046 mL) was added. The reaction mixture was slurried onto silica gel and directly purified by silica gel flash chromatography using 0-100% ethyl acetate in hexanes to give intermediate 3 (390 mg, 46% yield).

[0159] Step 3: To a solution of diol 3 (150 mg, 0.71 mmol, 1 equiv.) in dichloromethane (6 mL), acid 4 (579 mg, 1.6 mmol, 2.3 equiv.), EDCI hydrochloride (338 mg, 1.8 mmol, 2.5 equiv.), triethylamine (0.690 mL, 4.9 mmol, 7 equiv.), and 4-DMAP (17 mg, 0.14 mmol, 0.2 equiv.) were added at room temperature, and the resulting reaction mixture was stirred for 48 h. The reaction mixture was concentrated in vacuo and purified by silica gel flash chromatography using 0-25% ethyl acetate in hexanes to afford pure intermediate 5 (232 mg, 37% yield) as a clear, pale yellow syrup.

[0160] Step 4: Intermediate 5 (190 mg, 0.21 mmol, 1 equiv.) was added to a mixture of dichloromethane (9 mL) and water (0.565 mL). DDQ (122 mg, 0.53 mmol, 2.5 equiv.) was added slowly in portions, and the resulting reaction mixture was stirred at room temperature overnight. The reaction mixture was quenched by the addition of saturated aqueous sodium bicarbonate (15 mL). The aqueous layer was extracted with dichloromethane (4 × 25 mL). The combined organic layers were washed with saturated aqueous sodium bicarbonate (25 mL), brine (2 × 25 mL), dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure to give the crude product, which was purified by flash silica gel chromatography using 0–30% ethyl acetate in hexanes. Pure fractions were concentrated to dryness to give the desired product alcohol 6 (160 mg, quantitative) as a pale yellow, clear oil.

[0161] Step 5: To a solution of intermediate alcohol 6 (174 mg, 0.23 mmol, 1 equiv.) in dichloromethane (3.5 mL), Boc-Gly-Gly7 (80 mg, 0.34 mmol, 1.5 equiv.), EDCI hydrochloride (109 mg, 0.56 mmol, 2.5 equiv.), triethylamine (0.3 mL, 2.26 mmol, 10 equiv.), and 4-DMAP (14 mg, 0.11 mmol, 0.5 equiv.) were added, and the resulting reaction mixture was stirred at room temperature for 16 h. LCMS indicated the formation of the desired product. The reaction mixture was loaded directly onto a silica gel column and purified using 0–30% methanol in dichloromethane. Pure fractions were combined, concentrated on a rotovap, and dried under high vacuum overnight to afford the desired product 8 (200 mg, 90% yield). LCMS (M+1) 984.36, Rt=5.55 min (Method B).

[0162] Step 6: To a solution of intermediate 8 (105 mg, 0.11 mmol, 1 equiv.) in dichloromethane (5 mL) was added 0.5 mL of TFA. The resulting reaction mixture was stirred at room temperature for 30 minutes, then concentrated on a rotovap and dried under high vacuum to give the desired product (I) (88 mg, 80% yield). LCMS (M+1) 884.36, Rt = 4.98 min (Method B).

[0163] Step 7: To a solution of intermediate 8 (105 mg, 0.11 mmol, 1 equiv.) in dichloromethane (5 mL) was added 0.5 mL of HCl in dioxane, and the resulting reaction mixture was stirred at room temperature. After 30 minutes, the reaction mixture was concentrated in vacuo and dried under high vacuum to give the desired product (II). (87 mg, 86% yield). LCMS (M+1) 884.36, Rt = 4.98 min (Method B).

[0164] The synthesis of compound III is described in the following scheme. [ka]

[0165] Step 1: This reaction was carried out under an argon atmosphere, and all glassware was dried in an oven. To a solution of compound 6 (385 mg, 0.5 mmol, 1 equiv.) in dry DCM (5 mL) was added p-nitrobenzyl chloroformate (202 mg, 1.0 mmol, 2 equiv.). This solution was stirred at 0 °C for 20 min. 4-DMAP (141 mg, 1.15 mmol, 2.3 equiv.) was then added, and the resulting reaction mixture was stirred at room temperature for 16 h. The reaction mixture was concentrated on a rotovap, loaded directly onto a silica gel column, and purified using 0–30% ethyl acetate in hexanes. Pure fractions were combined, concentrated, and dried under high vacuum overnight to give compound 9 (375 mg, 80% yield) as a white solid.

[0166] Step 2: A solution of compound 9 (125 mg, 0.134 mmol, 1 equiv.) in anhydrous DCM (2 mL) was flushed with argon and stirred at 0 °C for 10 min. tert-Butyl-2-hydroxyethylaminoformylate 10 (87 mg, 0.535 mmol, 4 equiv.) was added, and the resulting reaction mixture was stirred at room temperature for 16 h. The reaction progress was monitored by LCMS. Upon completion of the reaction, it was loaded directly onto a silica gel column and purified using 0–40% acetone and hexanes. The pure fractions were combined, concentrated on a rotovap, and dried under high vacuum overnight to give compound 11 (35 mg, 27% yield) as a colorless liquid.

[0167] Step 3: To a solution of compound 11 (35 mg, 0.04 mmol, 1 equiv.) in DCM (2 mL) was added TFA (1 mL). The reaction mixture was stirred for 1 h and monitored by LCMS. Upon completion, the reaction mass was concentrated on a rotovap and dried under high vacuum overnight to give (III) (39 mg, quantitative) as a colorless liquid. LCMS (M+1) 857.5, Rt = 6.22 min (Method A).

[0168] The synthesis of compound IV is described in the following scheme. [ka]

[0169] Step 1: A solution of compound 9 (145 mg, 0.155 mmol, 1 equiv.) in anhydrous DCM (2 mL) was flushed with argon and stirred at 0 °C for 5 min. tert-Butyl-(2-hydroxyethyl)-N-methylaminoformylate 12 (109 mg, 0.62 mmol, 4 equiv.) was added, and the resulting reaction mixture was stirred at room temperature for 16 h. The reaction was complete as indicated by LCMS. The reaction mixture was loaded directly onto a silica gel column and purified using 0–40% acetone and hexanes. Pure fractions were combined, concentrated on a rotovap, and dried under high vacuum overnight to give compound 13 (142 mg, 94% yield) as a colorless liquid.

[0170] Step 2: To a solution of compound 13 (82 mg, 0.08 mmol, 1 equiv.) in DCM (2 mL) was added TFA (1 mL). The reaction mixture was stirred for 1 h and monitored by LCMS, which showed the reaction was complete. The reaction mixture was concentrated on a rotovap and dried under high vacuum overnight to give the desired product (IV) (79 mg, quantitative) as a colorless liquid. LCMS (M+1) 871.1, Rt = 6.75 min (Method A).

[0171] The synthesis of compounds V, VI, and VII is described in the following scheme. [ka]

[0172] General procedure: A solution of compound 9 (1 equiv.) in anhydrous DCM (0.032 M) was flushed with argon and stirred at 0 °C for 5 min. The appropriate dimethylamino alcohol (4 equiv.) was added, and the resulting reaction mixture was stirred at room temperature for 16 h. The reaction mixture was then loaded directly onto a silica gel column and purified using 0-40% acetone and hexanes. Pure fractions were combined, concentrated on a rotovap, and dried under high vacuum overnight to give the desired product. (V): (110 mg, 93% yield). LCMS (M+1) 885.21, Rt=4.92 min (Method A). (VI): (21 mg, 38% yield). LCMS (M+1) 899.5, Rt=5.09 min (Method A). (VII): (17 mg, 27% yield). LCMS (M+1) 913.25, Rt=5.03 min (Method A).

[0173] The synthesis of intermediate 4 is described below. [ka]

[0174] Step 1: To a solution of 2-hexyldecanoic acid (5 g, 19.5 mmol, 1 equiv.) in dichloromethane (120 mL), pentane-1,5-diol (6.1 g, 58.5 mmol, 3 equiv.), DCC (4.6 g, 22.2 mmol, 1.14 equiv.), and 4-DMAP (2.85 g, 23.4 mmol, 1.2 equiv.) were added. The reaction mixture was stirred at room temperature for 12 h. Hexane (100 mL) was added, the resulting precipitate was filtered off, and the filtrate was concentrated in vacuo to give the crude material, which was purified by silica gel chromatography using 0-15% ethyl acetate in hexane to give pure intermediate 17 (3.5 g, 52% yield).

[0175] Step 2: A solution of intermediate alcohol 17 (918 mg, 2.7 mmol, 1 equiv) in acetone (10 mL) was cooled to approximately 0 °C in an ice / water bath. Jones reagent (2 M, 2.16 mL, 4.32 mmol, 1.6 equiv) was added slowly via syringe, and the resulting reaction mixture was gradually warmed to room temperature and stirred for 16 h. Methanol (0.5 mL) was added to quench the reaction. The reaction mixture was concentrated under reduced pressure, diluted with water (200 mL), and extracted with ethyl acetate (100 mL). The aqueous layer was back-extracted with additional ethyl acetate (100 mL). The combined organic layers were dried over anhydrous sodium sulfate, filtered, and concentrated in vacuo. The crude product was purified by silica gel column chromatography using 0–15% ethyl acetate in hexanes to afford intermediate 4 (710 mg, 74% yield) as a clear, colorless oil. LCMS method: Method A: Chromolith, RP-18e, 50–4.6 mm, mobile phase A: 0.05% TFA in water, mobile phase B: 0.05% TFA in acetonitrile, 50A / 50B–0A / 100B over 8 min, hold 8–9 min, 95A / 5B 9–9.1 min, hold 9.1–12 min. Method B: Chromolith, RP-18e, 50–4.6 mm, mobile phase A: 0.05% TFA in water, mobile phase B: 0.05% TFA in acetonitrile, 50A / 50B–0A / 100B over 9.5 min, hold 9.5–10.5 min, 50A / 50B 10.5–12 min.

[0176] Abbreviation: N,N'-dicyclohexylcarbodiimide (DCC) Dichloromethane (DCM) 4-(Dimethylamino)pyridine (4-DAMP) 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide (EDCI) Triethylamine (TEA) 2,3-Dichloro-5,6-dicyano-1,4-benzoquinone (DDQ)

[0177] While several exemplary aspects and embodiments are discussed above, those skilled in the art will recognize certain modifications, permutations, additions, and subcombinations thereof. Accordingly, it is intended that the following appended claims and the claims hereafter written be interpreted to include all such modifications, permutations, additions, and subcombinations as fall within their true spirit and scope.

Claims

1. A compound having the following structure (I), or a pharmaceutically acceptable salt or stereoisomer thereof: 【Chemistry 1】 During the ceremony, R １ , R ２ , R ３ , and R ４ are each independently hydrogen, C １ ~C ２０ Alkyl, C １ ~C ２０ Alkynyl, C １ ~C ２０ alkenyl, or C １ ~C ２０ is heteroalkyl, Y １ and Y ２ However, each independently, C １ ~C ８ Alkyl, C １ ~C ８ Alkynyl, C １ ~C ８ Alkenyl, or aryl C １ ~C ６ is alkyl, X is -OR ５ or -CH ２ NR ６ R ７ and R ５ But C １ ~C ６ Alkyl or C １ ~C ６ is heteroalkyl, R ６ and R ７ are each independently hydrogen, C １ ~C ６ Alkyl, or —C(═O)R ８ and R ８ But C １ ~C ６ Alkyl, C １ ~C ６ Alkynyl, C １ ~C ６ alkenyl, or C １ ~C ６ It is heteroalkyl.

2. 10. The compound of claim 1 having one of the following structures (IA)-(IB), or a pharmaceutically acceptable salt or stereoisomer thereof: 【Chemistry 2】

3. R １ , R ２ , R ３ , and R ４ However, each independently, C １ ~C ２０ Alkyl, C １ ~C ２０ Alkynyl, or C １ ~C ２０ 3. The compound of claim 1 or 2, which is alkenyl.

4. R １ , R ２ , R ３ , and R ４ However, each independently, C １ ~C １８ Alkyl or C １ ~C １８ The compound of any one of claims 1 to 3, which is alkenyl.

5. R １ , R ２ , R ３ , and R ４ However, each independently, C ４ ~C １８ Alkyl or C ４ ~C １８ The compound of any one of claims 1 to 4, which is alkenyl.

6. R １ , R ２ , R ３ , and R ４ and each independently have one of the following structures: 【Transformation 3】

7. R １ , R ２ , R ３ , and R ４ and each independently have one of the following structures: 【Chemistry 4】

8. Y １ and Y ２ However, each independently, C １ ~C ６ Alkyl, C １ ~C ６ Alkynyl, C １ ~C ６ Alkenyl, or aryl C １ ~C ４ The compound of any one of claims 1 to 7, which is alkyl.

9. Y １ and Y ２ However, each independently, C １ ~C ６ Alkyl or C １ ~C ６ The compound of any one of claims 1 to 8, which is alkenyl.

10. Y １ and Y ２ However, each independently, C １ ~C ６ The compound of any one of claims 1 to 9, which is alkyl.

11. The compound of any one of claims 1 to 10, having one of the following structures (IC) to (ID), or a pharmaceutically acceptable salt or stereoisomer thereof: 【Transformation 5】 During the ceremony, n １ , n ２ , n ３ , and n ４ are each independently an integer from 0 to 19, m １ and m ２ is independently an integer from 1 to 8.

12. n １ , n ２ , n ３ , and n ４ The compound according to any one of claims 1 to 11, wherein each independently represents an integer from 0 to 17.

13. n １ , n ２ , n ３ , and n ４ is each independently an integer from 3 to 17.

14. m １ and m ２ The compound according to any one of claims 1 to 13, wherein each independently represents an integer from 1 to 6.

15. m １ and m ２ is independently an integer of 4.

16. R ５ But C １ ~C ４ Alkyl or C １ ~C ４ The compound of any one of claims 1 to 15, which is heteroalkyl.

17. R ５ But C １ ~C ４ Heteroalkyl or C ２ ~C ３ The compound of any one of claims 1 to 16, which is heteroalkyl.

18. R ５ C ２ ~C ３ Heteroalkyl is C １ ~C ６ Alkyl, C １ ~C ６ Alkynyl, C １ ~C ６ alkenyl, or C １ ~C ６ The compound of any one of claims 1 to 17, further substituted with heteroalkyl.

19. R ５ But C １ ~C ４ Amine or C ２ ~C ３ The compound of any one of claims 1 to 18, which is an amine.

20. R ５ but has the following structure: 【Transformation 6】 The compound according to any one of claims 1 to 19, wherein

21. R ６ and R ７ The compound of any one of claims 1 to 20, wherein each of is hydrogen.

22. R ６ and R ７ is hydrogen, and R ６ and R ７ The other of １ ~C ６ Alkyl or —C(═O)R ８ The compound according to any one of claims 1 to 21,

23. R ６ and R ７ is hydrogen, and R ６ and R ７ The other of the two is —C(═O)R ８ The compound according to any one of claims 1 to 22,

24. 24. The compound of claim 23, having the following structure (IE), or a pharmaceutically acceptable salt or stereoisomer thereof: 【Transformation 7】

25. R ８ But -CH ２ NH ２ or -CH ２ NHR ９ and R ９ But -CH ３ , -C ｚ H ｚ＋１ , or -(CH ２ ) ｚ 25. The compound of any one of claims 1 to 24, wherein Z is OH and Z is an integer from 1 to 6.

26. 10. The compound of claim 1 having one of the following structures: 【Transformation 8】 【change】 【change】 【change】 【change】 【change】 【change】 【change】 【change】

27. The compound according to any one of claims 1 to 26, wherein the compound is a trifluoroacetate salt or a hydrochloride salt.

28. A pharmaceutical composition comprising the compound of any one of claims 1 to 27 and a therapeutic agent comprising a nucleic acid.

29. 30. The pharmaceutical composition of claim 28, further comprising a helper lipid, a stabilizing lipid, or a structural lipid.

30. The helper lipid may be 1,2-dilinoleoyl-sn-glycero-3-phosphocholine (DLPC), 1,2-dimyristoyl-sn-glycero-phosphocholine (DMPC), 1,2-dioleoyl-sn-glycero-3-phosphocholine (DOPC), 1,2-dipalmitoyl-sn-glycero-3-phosphocholine (DPPC), 1,2-distearoyl-sn-glycero-3-phosphocholine (DSPC), 1,2-diundeca oleoyl-sn-glycero-phosphocholine (DUPC), 1-palmitoyl-2-oleoyl-sn-glycero-3-phosphocholine (POPC), 1,2-di-O-octadecenyl-sn-glycero-3-phosphocholine (18:0 diether PC), l-oleoyl-2-cholesterylhemisuccinoyl-sn-glycero-3-phosphocholine (OChemsPC), 1-hexadecyl-sn-glycero-3-phosphocholine (C16 Lyso PC), 1,2-dilinolenoyl-sn-glycero-3-phosphocholine, 1,2-diarachidonoyl-sn-glycero-3-phosphocholine, 1,2-didocosahexaenoyl-sn-glycero-3-phosphocholine, 1,2-dioleoyl-sn-glycero-3-phosphoethanolamine (DOPE), 1,2-diphytanoyl-sn-glycero-3-phosphoethanolamine (ME 16.0 PE), 1,2-distearoyl-sn-glycero-3-phosphoethanolamine, 1,2-dilinoleoyl-sn-glycero-3-phosphoethanolamine, 1,2-dilinolenoyl-sn-glycero-3-phosphoethanolamine, 1,2-diarachidonoyl-sn-glycero-3-phosphoethanolamine, 1,2-didocosahexaenoyl-sn-glycero-3-phosphoethanolamine, 1,2-dioleoyl-sn-glycero-3-phospho-rac-(1-glycerol) sodium salt (DOPG), and mixtures thereof.

31. 30. The pharmaceutical composition of claim 29, wherein the stabilizing lipid is 1-(monomethoxy-polyethylene glycol)-2,3-dimyristoylglycerol (PEG-DMG) with an average PEG molecular weight of 2000.

32. 30. The pharmaceutical composition of claim 29, wherein the structural lipid is selected from the group consisting of cholesterol, cholesterol derivatives, fecosterol, sitosterol, ergosterol, campesterol, stigmasterol, brassicasterol, tomatidine, ursolic acid, alpha-tocopherol, and mixtures thereof.

33. 29. The pharmaceutical composition of claim 28, wherein the nucleic acid is selected from small interfering RNA (siRNA), asymmetric interfering RNA (aiRNA), microRNA (miRNA), dicer substrate RNA (dsRNA), small hairpin RNA (shRNA), or messenger RNA (mRNA).

34. 34. The pharmaceutical composition of claim 33, wherein the nucleic acid is mRNA.

35. 30. A method for administering a therapeutic agent comprising a nucleic acid to a patient in need thereof, comprising administering to said patient the composition of claim 28.

36. A lipid nanoparticle comprising the compound of any one of claims 1 to 26 and a therapeutic agent comprising a nucleic acid.

37. The lipid nanoparticle of claim 36, wherein the therapeutic agent is mRNA.

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