Lipid nanoparticles for delivery of agents

Abstract

The present disclosure provides compositions capable of delivering a load (including therapeutic agents, e.g., RNA) to organs or tissues other than the liver, lung and spleen (e.g., brain, heart, kidney and muscular tissue). More specifically, the composition comprises a plurality of lipid particles comprising an agent, a cationic lipid, and an ionizable lipid.

Description

[0001] This application claims the benefit of U.S. Provisional Application No. 63 / 495,958, filed April 13, 2023. The entire teachings of the aforementioned application are incorporated herein by reference. Background of the Invention Nucleic acid-based therapeutics represent an emerging and promising class of drugs with the potential to provide a wide range of treatments for many currently untreatable diseases. Lipid nanoparticles (LNPs) are self-assembled nanostructures that encapsulate and deliver nucleic acids. Liver-targeted LNPs encapsulating therapeutic small interfering RNA (siRNA) have been clinically approved for the treatment of patients with transthyretin-mediated amyloidosis (ONPATTRO™). The incorporation of cationic lipids into liver-targeted LNPs has been shown to have the potential to redirect LNPs to the lungs and spleen. However, despite significant efforts to expand the scope of intravenous LNP administration, delivering LNPs to organs other than the liver, lungs, and spleen remains a challenge.

[0003] Therefore, there is still a need for compositions that can deliver the drug to organs other than the liver, lungs and / or spleen, or to other organs other than the liver, lungs and / or spleen. Summary of the Invention

[0004] This disclosure is based, in part, on the discovery that the composition is capable of delivering pharmaceutical agents, particularly therapeutic agents, to organs or tissues other than the liver, lungs, and / or spleen.

[0005] Some embodiments provide compositions comprising a plurality of lipid nanoparticles, wherein the lipid nanoparticles comprise an agent, an ionizable lipid, and a fully saturated cationic lipid, wherein the lipid nanoparticles are free of zwitterionic phospholipids.

[0006] Some embodiments provide compositions comprising a plurality of lipid nanoparticles, wherein the lipid nanoparticles comprise an agent, an ionizable lipid, and a cationic lipid, wherein the cationic lipid is a lipid according to formula VI: Where the variable (e.g., R) 101 R 102 R 105 The values ​​of (X) are as defined in this article.

[0007] Some embodiments provide compositions comprising a plurality of lipid nanoparticles, wherein the lipid nanoparticles comprise a pharmaceutical agent, an ionizable lipid, and a cationic lipid, wherein the ionizable lipid is a lipid of formula I or a pharmaceutically acceptable salt thereof: Among them, variables (e.g., X, R) 2 R 3 R4 R 5 R 6 R 7 R 8 R 9 R 10 R 11 R 12 The values ​​of a, b, c, d) are as defined in this paper.

[0008] Some embodiments provide compositions comprising a plurality of lipid nanoparticles, wherein the lipid nanoparticles comprise a pharmaceutical agent, an ionizable lipid, and at least one of EDPPC and DPTAP, wherein the ionizable lipid is a lipid of formula I or II or a pharmaceutically acceptable salt thereof. Where the variable (e.g., R) 51 R 52 L 10 G 30 、a'、L 30 G 10 L 20 b' R 53 R 54 R 57 The value of ) is as defined in this article.

[0009] In some embodiments, this disclosure further provides a method for delivering a medicament to one or more of the brain, heart, muscles, and kidneys of an individual in need, the method comprising systemically administering to the individual a composition comprising a plurality of lipid nanoparticles, wherein the lipid nanoparticles comprise the medicament, ionizable lipids, and fully saturated cationic lipids.

[0010] Some embodiments provide a method for delivering a pharmaceutical agent to an individual in need, the method comprising administering the composition disclosed herein to the individual.

[0011] Some embodiments provide methods for treating an individual suffering from a disease, disorder, or condition that is beneficially treatable by a pharmaceutical agent, the methods comprising administering to the individual a therapeutically effective amount of a composition comprising the pharmaceutical agent of the present disclosure.

[0012] In some embodiments, this document also provides the use of the compositions of this disclosure according to the methods described herein, for example, the use of the compositions of this disclosure comprising a pharmaceutical agent for delivering the pharmaceutical agent to an individual (e.g., an individual in need); the use of the compositions of this disclosure comprising a pharmaceutical agent for treating an individual suffering from a disease, disorder, or condition that is beneficially treatable by the pharmaceutical agent. In some embodiments, this document also provides the use of the compositions of this disclosure in the preparation of a medicament used in the methods described herein, for example, delivering a pharmaceutical agent to an individual (e.g., an individual in need); treating an individual suffering from a disease, disorder, or condition that is beneficially treatable by the pharmaceutical agent. In some embodiments, this document also provides the use of the compositions according to the methods described herein, for example, delivering a pharmaceutical agent to an individual (e.g., an individual in need); treating a disease, disorder, or condition that is beneficially treatable by the pharmaceutical agent.

[0013] Surprisingly, it has been found that by incorporating specific cationic lipids into LNPs targeting hepatocytes at an optimized molar percentage, these LNPs can deliver RNA to a variety of organs, including brain and muscle endothelial cells, in addition to the liver, lungs, and spleen. Furthermore, by adjusting the ratio of these cationic lipids in the LNPs, RNA delivery to organs other than the liver and lungs has been successfully achieved. The cationic lipid 1,2-dipalmitoyl- was identified through rigorous formulation screening. sn -glycerol-O-ethyl-3-phosphorylcholine (EDPPC), 1,2-dipalmitoyl-3-trimethylammonium-propane (DPTAP), and O,O'-bis(tetradecanoyl)-N-(α-trimethylammoniumacetyl)diethanolamine (DC-6-14) are more potent than 1,2-dioleoyl-3-trimethylammonium-propane (DOTAP) and 1,2-dimyristoyl- sn 1-Glyceryl-O-ethyl-3-phosphorylcholine (EDMPC) delivers RNA more efficiently to endothelial cells. Attached Figure Description

[0014] Figure 1 The names and structures of several ionizable lipids and multiple classes of ionizable lipids applicable to this disclosure are shown.

[0015] Figure 2A-2G This demonstrates the results of replacing zwitterionic phospholipids with cationic lipids. In this experiment, lipids encapsulated with VE-cadherin (VE-cadherin) were used. Cdh5 The siRNA LNP was administered intravenously to mice at 0.5 mg / kg, and organs were harvested 72 hours after injection.

[0016] After isolating and purifying total RNA from organs, RT-qPCR was used to compare it with housekeeping genes. Gusb Quantitative VE-cadherin ( Cdh5 mRNA. Figure 2AThe data from this experiment in the liver are presented. Figure 2B The data from this experiment in the spleen are presented. Figure 2C The data from this experiment in the kidneys are presented. Figure 2D This demonstrates the data from the experiment in the lungs. Figure 2E The data from this experiment in the heart are presented. Figure 2F This demonstrates the data from the experiment in the brain. Figure 2G The data from this experiment in muscle are presented.

[0017] Figures 3A-3E This demonstrates a comparison of LNP containing 20% ​​EDOPC versus (vs.) 50% DOTAP for delivery to selected organs. The LNP will be encapsulated with VE-cadherin (…). Cdh5 The LNP of the siRNA was administered intravenously to mice at 0.3 mg / kg, and organs were harvested 72 hours post-injection. Total RNA was isolated and purified from the organs, and its expression was compared with housekeeping genes using RT-qPCR. Gusb Quantitative VE-cadherin ( Cdh5 mRNA. Figure 3A The data from this experiment in the liver are presented. Figure 3B The data from this experiment in the spleen are presented. Figure 3C The data from this experiment in the kidneys are presented. Figure 3D This demonstrates the data from the experiment in the lungs. Figure 3E The data from this experiment in the heart are presented.

[0018] Figure 4A-4G Screening results for LNPs containing 20 mol% cationic lipids in selected organs are presented. Mice were intravenously administered a formulation containing siRNA targeting VE-cadherin at 0.5 mg / kg, and organs were harvested 72 hours post-injection. Total RNA was isolated and purified from the organs, relative to housekeeping genes. Gusb Quantitative VE-cadherin ( Cdh5 mRNA was also tested. A PBS control was also tested. Figure 4A The data from this experiment in the liver are presented. Figure 4B The data from this experiment in the spleen are presented. Figure 4C The data from this experiment in the kidneys are presented. Figure 4D This demonstrates the data from the experiment in the lungs. Figure 4E The data from this experiment in the heart are presented. Figure 4F This demonstrates the data from the experiment in the brain. Figure 4G The data from this experiment in muscle are presented.

[0019] Figure 5A-5G This study presents a comparison of LNPs containing 20 mol% EDPPC and EDMPC, using MC3 and MD1 as ionizable lipids. The LNPs are encapsulated with targeted... Cdh5The siRNA LNP was administered intravenously to mice at 0.5 mg / kg, and organs were harvested 72 hours post-injection. Total RNA was isolated and purified from the organs, relative to housekeeping genes. Gusb Quantitative Cdh5 mRNA. Figure 5A The data from this experiment in the liver are presented. Figure 5B The data from this experiment in the spleen are presented. Figure 5C The data from this experiment in the kidneys are presented. Figure 5D This demonstrates the data from the experiment in the lungs. Figure 5E The data from this experiment in the heart are presented. Figure 5F This demonstrates the data from the experiment in the brain. Figure 5G The data from this experiment in muscle are presented.

[0020] Figure 6A-6G This study presents a comparison of LNPs containing 20 mol% DPTAP and DOTAP, using MC3 and MD1 as ionizable lipids. The LNPs are encapsulated with targeted DPTAP and DOTAP. Cdh5 The siRNA LNP was administered intravenously to mice at 0.5 mg / kg, and organs were harvested 72 hours post-injection. Total RNA was isolated and purified from the organs, relative to housekeeping genes. Gusb Quantitative Cdh5 mRNA. Figure 6A The data from this experiment in the liver are presented. Figure 6B The data from this experiment in the spleen are presented. Figure 6C The data from this experiment in the kidneys are presented. Figure 6D This demonstrates the data from the experiment in the lungs. Figure 6E The data from this experiment in the heart are presented. Figure 6F This demonstrates the data from the experiment in the brain. Figure 6G The data from this experiment in muscle are presented.

[0021] Figures 7A-7D This demonstrates the improvement in RNA delivery when using a combination of ionizable and cationic lipids compared to using ionizable and cationic lipids alone. The encapsulation of targeted... Cdh5 The siRNA LNP was administered intravenously to mice at 0.3 mg / kg, and organs were harvested 72 hours post-injection. Total RNA was isolated and purified from the organs, relative to housekeeping genes. Gusb Quantitative Cdh5 mRNA. Figure 7A The data from this experiment in the liver are presented. Figure 7B This demonstrates the data from the experiment in the lungs. Figure 7C The data from this experiment in the heart are presented. Figure 7D The data from this experiment in muscle are presented.

[0022] Figure 8A-8GThis demonstrates the results of combining 20 mol% EDPPC with various ionizable lipids. The encapsulated lipids were targeted... Cdh5 The siRNA formulation was administered intravenously to mice at 0.5 mg / kg, and organs were harvested 72 hours post-injection. Total RNA was isolated and purified from the organs, relative to housekeeping genes. Gusb Quantitative analysis of VE-cadherin mRNA was performed. A PBS control was also tested. Figure 8A The data from this experiment in the liver are presented. Figure 8B The data from this experiment in the spleen are presented. Figure 8C The data from this experiment in the kidneys are presented. Figure 8D This demonstrates the data from the experiment in the lungs. Figure 8E The data from this experiment in the heart are presented. Figure 8F This demonstrates the data from the experiment in the brain. Figure 8G The data from this experiment in muscle are presented.

[0023] Figure 9A-9G This demonstrates the results of combining 20 mol% EDPPC with various ionizable lipids. The encapsulated lipids were targeted... Cdh5 The siRNA formulation was administered intravenously to mice at 0.5 mg / kg, and organs were harvested 72 hours post-injection. Total RNA was isolated and purified from the organs, relative to housekeeping genes. Gusb Quantitative analysis of VE-cadherin mRNA was performed. A PBS control was also tested. Figure 9A The data from this experiment in the liver are presented. Figure 9B The data from this experiment in the spleen are presented. Figure 9C The data from this experiment in the kidneys are presented. Figure 9D This demonstrates the data from the experiment in the lungs. Figure 9E The data from this experiment in the heart are presented. Figure 9F This demonstrates the data from the experiment in the brain. Figure 9G The data from this experiment in muscle are presented.

[0024] Figure 10A-10G This demonstrates the results of combining 20 mol% EDPPC with various ionizable lipids. The encapsulated lipids were targeted... Cdh5 The siRNA formulation was administered intravenously to mice at 0.5 mg / kg, and organs were harvested 72 hours post-injection. Total RNA was isolated and purified from the organs, relative to housekeeping genes. Gusb Quantitative analysis of VE-cadherin mRNA was performed. A PBS control was also tested. Figure 10A The data from this experiment in the liver are presented. Figure 10B The data from this experiment in the spleen are presented. Figure 10C The data from this experiment in the kidneys are presented. Figure 10D This demonstrates the data from the experiment in the lungs. Figure 10E The data from this experiment in the heart are presented. Figure 10F This demonstrates the data from the experiment in the brain. Figure 10G The data from this experiment in muscle are presented.

[0025] Figure 11A-11G Results of combining 20 mol% DPTAP with various ionizable lipids are presented. Mice were intravenously administered a formulation containing siRNA targeting VE-cadherin at 0.5 mg / kg, and organs were harvested 72 hours post-injection. Total RNA was isolated and purified from the organs, relative to housekeeping genes. Gusb Quantitative analysis of VE-cadherin mRNA was performed. A PBS control was also tested. Figure 11A The data from this experiment in the liver are presented. Figure 11B The data from this experiment in the spleen are presented. Figure 11C The data from this experiment in the kidneys are presented. Figure 11D This demonstrates the data from the experiment in the lungs. Figure 11E The data from this experiment in the heart are presented. Figure 11F This demonstrates the data from the experiment in the brain. Figure 11G The data from this experiment in muscle are presented.

[0026] Figure 12A-12G Results of combining 20 mol% DPTAP with various ionizable lipids are presented. Mice were intravenously administered a formulation containing siRNA targeting VE-cadherin at 0.5 mg / kg, and organs were harvested 72 hours post-injection. Total RNA was isolated and purified from the organs, relative to housekeeping genes. Gusb Quantitative analysis of VE-cadherin mRNA was performed. A PBS control was also tested. Figure 12A The data from this experiment in the liver are presented. Figure 12B The data from this experiment in the spleen are presented. Figure 12C The data from this experiment in the kidneys are presented. Figure 12D This demonstrates the data from the experiment in the lungs. Figure 12E The data from this experiment in the heart are presented. Figure 12F This demonstrates the data from the experiment in the brain. Figure 12G The data from this experiment in muscle are presented.

[0027] Figure 13A-13G Results are presented for 20 mol% DC-6-14 combined with various ionizable lipids. Mice were intravenously administered a formulation containing siRNA targeting VE-cadherin at 0.5 mg / kg, and organs were harvested 72 hours post-injection. Total RNA was isolated and purified from the organs, relative to housekeeping genes. Gusb Quantitative analysis of VE-cadherin mRNA was performed. A PBS control was also tested. Figure 13A The data from this experiment in the liver are presented. Figure 13B The data from this experiment in the spleen are presented. Figure 13C The data from this experiment in the kidneys are presented. Figure 13D This demonstrates the data from the experiment in the lungs. Figure 13E The data from this experiment in the heart are presented. Figure 13F This demonstrates the data from the experiment in the brain. Figure 13G The data from this experiment in muscle are presented.

[0028] Figure 14A-14G Results are presented for 20 mol% DC-6-14 combined with various ionizable lipids. Mice were intravenously administered a formulation containing siRNA targeting VE-cadherin at 0.5 mg / kg, and organs were harvested 72 hours post-injection. Total RNA was isolated and purified from the organs, relative to housekeeping genes. Gusb Quantitative analysis of VE-cadherin mRNA was performed. A PBS control was also tested. Figure 14A The data from this experiment in the liver are presented. Figure 14B The data from this experiment in the spleen are presented. Figure 14C The data from this experiment in the kidneys are presented. Figure 14D This demonstrates the data from the experiment in the lungs. Figure 14E The data from this experiment in the heart are presented. Figure 14F This demonstrates the data from the experiment in the brain. Figure 14G The data from this experiment in muscle are presented.

[0029] Figure 15A-15G Experimental results demonstrating the optimized ratio of EDPPC in LNPs are presented. To optimize the ratio of the cationic lipids used for extrahepatic delivery, a series of LNPs with a systematically increased percentage of EDPPC were prepared and tested in vivo with a single administration of 0.5 mg / kg siCdh5. LNPs containing 10%–50% EDPPC showed efficacy in the liver, spleen, kidney, lung, and brain. Cdh5 Gene silencing. LNPs containing 20%–30% EDPPC showed the most effective gene silencing in the heart and skeletal muscle. A PBS control was also tested. Figure 15A The data from this experiment in the liver are presented. Figure 15B The data from this experiment in the spleen are presented. Figure 15C The data from this experiment in the kidneys are presented. Figure 15D This demonstrates the data from the experiment in the lungs. Figure 15E The data from this experiment in the heart are presented. Figure 15F This demonstrates the data from the experiment in the brain. Figure 15G The data from this experiment in muscle are presented.

[0030] Figure 16A-16GExperimental results demonstrating the optimization of the DPTAP ratio in LNPs are presented. To optimize the ratio of the cationic lipids used for extrahepatic delivery, a series of LNPs with a systematically increased percentage of DPTAP were prepared and tested in vivo with a single administration of 0.5 mg / kg siCdh5. LNPs containing 10%–50% DPTAP showed efficacy in the liver, spleen, kidney, lung, heart, and skeletal muscle. Cdh5 Gene silencing. A PBS control was also tested. Figure 16A The data from this experiment in the liver are presented. Figure 16B The data from this experiment in the spleen are presented. Figure 16C The data from this experiment in the kidneys are presented. Figure 16D This demonstrates the data from the experiment in the lungs. Figure 16E The data from this experiment in the heart are presented. Figure 16F This demonstrates the data from the experiment in the brain. Figure 16G The data from this experiment in muscle are presented.

[0031] Figure 17A-17G Experimental results showcasing the optimized DC-6-14 ratio in LNPs. To optimize the ratio of the cationic lipids for extrahepatic delivery, a series of LNPs with a systematically increased percentage of DC-6-14 were prepared and tested in vivo with a single administration of 0.5 mg / kg siCdh5. LNPs containing 10%–50% DC-6-14 showed efficacy in the liver, spleen, kidney, lung, heart, and skeletal muscle. Cdh5 Gene silencing. A PBS control was also tested. Figure 17A The data from this experiment in the liver are presented. Figure 17B The data from this experiment in the spleen are presented. Figure 17C The data from this experiment in the kidneys are presented. Figure 17D This demonstrates the data from the experiment in the lungs. Figure 17E The data from this experiment in the heart are presented. Figure 17F This demonstrates the data from the experiment in the brain. Figure 17G The data from this experiment in muscle are presented.

[0032] Figure 18 This study demonstrates the efficacy of siRNA delivery to the lungs when combined with liver-targeted ionizable lipids (LNPs) and DOTAP. To investigate whether adding 50% DOTAP to liver-targeted LNPs containing ionizable lipids of formula I or III redirected them to the lungs, five LNPs with different ionizable lipids (MC3 as a positive control) and one LNP without ionizable lipids as a negative control were prepared. Their in vivo endothelial delivery efficiency was tested using a single intravenous administration of 0.4 mg / kg siCdh5. Data from this experiment are presented.

[0033] Figures 19A-19EThis study presents the tissue tropism results obtained by altering the alkyl chain length of PEG lipids in LNPs. To investigate whether the alkyl chain length of PEG lipids affects the tissue tropism of LNPs containing 50% DOTAP, three LNPs with different PEG lipids were prepared and their endothelial delivery efficiency was tested in vivo using a single intravenous administration of 1.0 mg / kg siCdh5. Tissue samples were obtained 3 days after administration, and quantification was performed by RT-qPCR. Cdh5 mRNA. All the LNPs mentioned showed comparable results in all collected tissues. Cdh5 Gene silencing. Figure 19A The data from this experiment in the liver are presented. Figure 19B The data from this experiment in the spleen are presented. Figure 19C The data from this experiment in the kidneys are presented. Figure 19D This demonstrates the data from the experiment in the lungs. Figure 19E The data from this experiment in the heart are presented.

[0034] Figure 20A-20G Results of combining 20 mol% EDOPC with various ionizable lipids are presented. To explore the general applicability of EDOPC for extrahepatic delivery, various ionizable lipids were formulated with 20% EDOPC. LNPs with five different ionizable lipids were prepared, and their endothelial delivery efficiency was tested in vivo using a single intravenous administration of 0.5 mg / kg siCdh5. Tissue samples were obtained 3 days after administration, and quantification was performed by RT-qPCR. Cdh5 mRNA. All LNPs containing 20% ​​EDOPC showed significant activity in all tested tissues. Cdh5 Gene silencing indicates that these new formulations can be used for RNA delivery to endothelial RNA in a variety of tissues. Figure 20A The data from this experiment in the liver are presented. Figure 20B The data from this experiment in the spleen are presented. Figure 20C The data from this experiment in the kidneys are presented. Figure 20D This demonstrates the data from the experiment in the lungs. Figure 20E The data from this experiment in the heart are presented. Figure 20F This demonstrates the data from the experiment in the brain. Figure 20G The data from this experiment in muscle are presented.

[0035] Figures 21A-21EThis study demonstrates tissue tropism results by altering the alkyl tail length of PEG lipids in LNPs containing 18.5% EDOPC. To investigate whether the alkyl chain length of PEG lipids affects the tissue tropism of LNPs containing 18.5% EDOPC, three LNPs with different PEG lipids were prepared and their endothelial delivery efficiency was tested in vivo using a single intravenous administration of 1.0 mg / kg siCdh5. Tissue samples were obtained 3 days after administration, and quantification was performed by RT-qPCR. Cdh5 mRNA. All the LNPs mentioned showed comparable results in all collected tissues. Cdh5 Gene silencing. Figure 21A The data from this experiment in the liver are presented. Figure 21B The data from this experiment in the spleen are presented. Figure 21C The data from this experiment in the kidneys are presented. Figure 21D This demonstrates the data from the experiment in the lungs. Figure 21E The data from this experiment in the heart are presented.

[0036] Figures 22A-22F Results of combining 10 mol% DPTAP with various ionizable lipids are presented. To explore the general applicability of DPTAP for extrahepatic delivery, various ionizable lipids were formulated with 10% DPTAP. LNPs with six different ionizable lipids were prepared, and their endothelial delivery efficiency was tested in vivo using a single intravenous administration of 0.5 mg / kg siCdh5. Tissue samples were obtained 3 days after administration, and quantification was performed by RT-qPCR. Cdh5 mRNA. All LNPs containing 10% DPTAP showed significant activity in all tested tissues. Cdh5 Gene silencing indicates that these new formulations can be used for RNA delivery to endothelial RNA in a variety of tissues. Figure 22A The data from this experiment in the liver are presented. Figure 22B The data from this experiment in the kidneys are presented. Figure 22C This demonstrates the data from the experiment in the lungs. Figure 22D The data from this experiment in the heart are presented. Figure 22E This demonstrates the data from the experiment in the brain. Figure 22F The data from this experiment in muscle are presented. Detailed Implementation

[0037] Unless otherwise defined, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which this disclosure pertains. Although similar or equivalent methods and materials to those described herein may be used according to this disclosure, suitable methods and materials are described below. All publications, patent applications, patents, and other references mentioned herein are incorporated herein by reference in their entirety. Furthermore, the materials, methods, and examples described are illustrative only and not intended to be limiting. Other features and advantages of this disclosure will become apparent in the following detailed description and claims.

[0038] A. Definition The compounds described herein include any and all possible isomers, stereoisomers, enantiomers, diastereomers, tautomers, salts (e.g., pharmaceutically acceptable salts), and solvates thereof. Therefore, as used in this disclosure, the term "compound" refers to any one of the compounds of this disclosure and all possible isomers, stereoisomers, enantiomers, diastereomers, tautomers, pharmaceutically acceptable salts, and solvates thereof. Unless otherwise specifically indicated in the text or by the use of standard notation in the art to describe a particular regioisomer, stereoisomer, epiisomer, enantiomer, or configuration, such references apply.

[0039] The compounds described herein may have an asymmetric center, a chiral axis, and a chiral plane (e.g., as described in ELEliel and SH Wilen, Stereochemistry of Carbon Compounds, John Wiley & Sons, New York, 1994, pp. 1119-1190), and may exist as racemic mixtures, individual isomers (e.g., diastereomers, enantiomers, geometric isomers (including cis and trans double bond isomers), conformational isomers (including rotational and steganous isomers), tautomers, and mixtures of intermediates, including all possible isomers and mixtures thereof, unless otherwise stated.

[0040] When a compound is described by structure without indicating stereochemistry, and the compound has one or more chiral centers, it should be understood that the structure encompasses an enantiomer or diastereomer of the compound that is separate from or substantially separate from its corresponding optical isomer, a racemic mixture of the compound, and a mixture rich in one enantiomer or diastereomer relative to its corresponding optical isomer. When a compound is described by a structure indicating stereochemistry, and the compound has more than one chiral center, the stereochemistry indicates relative stereochemistry, not the absolute configuration of the substituents surrounding one or more chiral carbon atoms. Unless otherwise stated herein, when a compound is described by a structure indicating stereochemistry using solid and / or dashed wedges, and the compound has one or more chiral centers, the stereochemistry indicates the absolute configuration of the substituents surrounding the one or more chiral centers. Unless otherwise stated herein, when a compound is described by a structure indicating stereochemistry using bold solid and / or dashed lines, and the compound has one or more chiral centers, the stereochemistry indicates the relative stereochemistry of the unspecified absolute configuration of the substituents surrounding the one or more chiral centers. “R” and “S” can also be used, or optionally, to indicate the absolute configuration of substituents around one or more chiral centers (e.g., carbon atoms).

[0041] Therefore, for example, a single stereoisomer with known relative and absolute configurations having two chiral centers can be identified using the conventional RS system (e.g., (1S,2S)); diastereomers in racemic mixtures can be identified using an RS system with two letters (e.g., (1RS,2RS) as a racemic mixture of (1R,2R) and (1S,2S); (1RS,2SR) as a racemic mixture of (1R,2S) and (1S,2R)).

[0042] The English articles “a” and “an” are used in this disclosure to refer to one or more (i.e., at least one) grammatical objects of the article. For example, “element” means one or more elements. Similarly, when introducing elements disclosed herein, the English articles “a,” “an,” “the,” and “described” are intended to indicate the presence of one or more elements. Furthermore, the one or more elements may be the same or different.

[0043] Unless otherwise stated, the term “or” is used in this disclosure to mean the term “and / or” and is used interchangeably with the term “and / or”.

[0044] The term "about" means within an acceptable range of error for a given value, as determined by one of ordinary skill in the art. Typically, the acceptable range of error for a given value depends at least in part on how the value is measured or determined, for example, limitations of the measurement system. For instance, according to practice in the art, "about" may mean within an acceptable standard deviation. Alternatively, "about" may mean a range of ±20% of a given value, such as ±10%, ±5%, or ±1%. It should be understood that the term "about" may precede any specific value specified herein, except for the specific values ​​used in the embodiments.

[0045] Unless otherwise specifically defined, "alkyl" refers to a straight-chain or branched hydrocarbon group that is fully saturated and may include a divalent group, having one to about 30 carbon atoms if saturated, or two to about 30 carbon atoms if unsaturated. Examples of saturated alkyl groups include, but are not limited to, groups such as methyl (Me), ethyl (Et), n-propyl, isopropyl, n-butyl, tert-butyl, isobutyl, sec-butyl, and isomers such as n-pentyl, n-hexyl, n-heptyl, n-octyl, 1,1-dimethyl-heptyl, and 1,2-dimethyl-heptyl. Unsaturated alkyl groups may include one or more double bonds, triple bonds, or combinations thereof. Examples of unsaturated alkyl groups include, but are not limited to, vinyl, propenyl, isopropenyl, crotonyl, 2-isopentenyl, allenyl, butenyl, butadienyl, pentenyl, pentadienyl, 3-(1,4-pentadienyl), hexenyl, hexadienyl, ethynyl, propynyl, butynyl, and other isomers. Unless otherwise specifically defined, the term "divalent alkyl group" refers to the general formula: -alkyl-. A divalent alkyl group is an alkyl group having two connection points with a larger structural and / or other substituents. The term "C 1-n "Alkyl" refers to an alkyl group having 1 to n carbon atoms, where n is an integer greater than 1.

[0046] "Alkenyl" refers to a branched or straight-chain aliphatic group having a specified number of carbon atoms and at least one (e.g., one, two, three, four, five, etc.) carbon-carbon double bond. Therefore, "(C1-C..." 15 "Alkenyl" refers to a group having 1-15 carbon atoms, at least one carbon-carbon double bond, and arranged in a branched or straight chain. In some embodiments, the alkenyl group is (C1-C1) 30 )alkenyl, for example (C1-C 20 )alkenyl, (C1-C 15 )alkenyl, (C1-C 10 Alkenyl, (C1-C5)alkenyl, or (C1-C3)alkenyl. Examples of alkyl groups include vinyl, allyl, etc. In some embodiments, the alkenyl group is optionally substituted with one or more substituents, such as those described herein.

[0047] The term "alkynyl" refers to a branched or straight-chain aliphatic group having a carbon atom as a linker and at least one carbon-carbon triple bond. As used herein, the term alkynyl does not exclude the presence of one or more non-aromatic carbon-carbon double bonds. The groups -C≡CH, -C≡CCH3, and -CH2C≡CCH3 are non-limiting examples of alkynyl groups. "Alkyne" refers to a class of compounds having the formula HR, where R is an alkynyl group.

[0048] As used herein, the term "hydrocarbon" refers to an aliphatic moiety containing carbon and hydrogen. "Hydrocarbon group" should be understood as an alkyl, alkenyl, or alkynyl group or a combination thereof.

[0049] "Alkoxy" refers to an alkyl group linked by an oxygen atom, wherein the alkyl group is as described herein. Examples of alkoxy groups include, but are not limited to, methoxy, ethoxy, propoxy, and isopropoxy.

[0050] "Aryl" refers to a monocyclic or polycyclic (e.g., bicyclic, tricyclic) carbocyclic or aromatic ring system with a specified number of ring atoms, and includes fused to a non-aromatic aromatic ring, provided that one of the fused rings is an aromatic ring composed of hydrogen and carbon. Therefore, "(C6-C 15 "Aryl" refers to an aromatic ring system having 6-15 ring atoms. In some embodiments, the aryl group is (C6-C6) 25 ) aryl, for example (C6-C 20 )Aryl, (C6-C 15 )Aryl, (C6-C 12 ) aryl or (C6-C 10 Aryl. Examples of aryl groups include phenyl, naphthyl, and fluorenyl. In some embodiments, the aryl group is phenyl or fluorenyl.

[0051] The term "heteroaryl" refers to a monovalent aromatic group having an aromatic carbon or nitrogen atom as a linker, said carbon or nitrogen atom forming part of one or more aromatic ring structures, wherein at least one ring atom is nitrogen, oxygen, or sulfur, and said heteroaryl group is not composed of atoms other than carbon, hydrogen, aromatic nitrogen, aromatic oxygen, and aromatic sulfur. A heteroaryl ring may contain 1, 2, 3, or 4 ring atoms selected from nitrogen, oxygen, and sulfur. If more than one ring is present, then the rings may be fused or unfused. As used herein, the term does not exclude the presence of one or more alkyl, aryl, and / or aralkyl groups (as permitted by carbon number restrictions) connected to an aromatic ring or aromatic ring system. Non-limiting examples of heteroaryl groups include furanyl, imidazolyl, indolyl, indazole, isoxazolyl, methylpyridinyl, oxazolyl, phenylpyridinyl, pyridinyl, pyrroleyl, pyrimidinyl, pyrazinyl, quinolinyl, quinazolinyl, quinoxalinyl, triazinyl, tetrazolyl, thiazolyl, thiopheneyl, and triazolyl. The term "heteroaryl" can also refer to a divalent aromatic group having two aromatic carbon atoms, two aromatic nitrogen atoms, or one aromatic carbon atom and one aromatic nitrogen atom as two connecting points, said atoms forming part of one or more aromatic ring structures, wherein at least one of said ring atoms is nitrogen, oxygen, or sulfur, and said divalent group is not composed of atoms other than carbon, hydrogen, aromatic nitrogen, aromatic oxygen, and aromatic sulfur. If more than one ring is present, then the rings can be fused or unfused. Unfused rings can be connected by one or more of the following: covalent bonds, alkyl or alkenyl groups (carbon number restrictions allow). As used herein, the term does not exclude the presence of one or more alkyl, aryl, and / or aralkyl groups attached to an aromatic ring or aromatic ring system (carbon number restrictions allow).

[0052] Unless otherwise specifically defined, the term "cycloalkyl" refers to a fully saturated cyclic alkyl group containing one to four rings, each ring having three to eight carbon atoms. Exemplary examples of such groups include, but are not limited to, cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, cycloheptyl, adamantyl, bicyclo[1.1.1]pentane, etc.

[0053] Generally, "substituted" or "optionally substituted" means that in a group (e.g., an alkyl group, an aryl group), one or more bonds bonded to a hydrogen atom contained therein can be replaced by bonds bonded to non-hydrogen or non-carbon atoms. Substituted groups also include groups in which one or more bonds bonded to a carbon or hydrogen atom are replaced by one or more bonds bonded to a heteroatom (including double or triple bonds). Therefore, unless otherwise stated, a substituted group is replaced by one or more substituents, which can be the same or different. In some embodiments, the substituted group is replaced by 1, 2, 3, 4, 5, or 6 substituents. It should be understood that "substituted" or "replaced by" includes the implicit condition that the substitution is in accordance with the permissible valence of the substituted atom. Examples of substituents include, but are not limited to, alkyl, alkynyl, alkenyl, aryl, heteroaryl, aralkyl, alkylaryl, alcohols, halogens, hydroxyl; alkoxy, alkenyloxy, alkynoxy, aryloxy, arylalkoxy, heterocyclic alkoxy groups, carbonyl, carboxyl, esters, ethyl carbamate, oxime, hydroxylamine, alkoxyamine, arylalkoxyamine, thiols, sulfides, sulfoxides, sulfones, sulfonyl groups, sulfonamides, sulfonates, and sulfates. (lfate), amine, N-oxide, hydrazine, acylhydrazine, hydrazone, azide, amide, urea, amidine, guanidine, enamine, diimide, isocyanate, isothiocyanate, cyanate, thiocyanate, imine, nitro, nitrile, =CF2, -CF3, -CF2H, =CH2, -CHO, C(halogen)3, -ONO2, -alkyl-ONO2, -C(O)OP(O)(O-alkyl)2, -OPO3H2 and polyethylene glycol.

[0054] "Hydroxy group" refers to -OH.

[0055] Unless otherwise specifically defined, "alkylamino" refers to the general formula -(NH)-alkyl. Unless otherwise specifically defined, "dialkylamino" refers to the general formula -N-(alkyl)2. Unless otherwise specifically defined, dialkylamino includes cyclic amine compounds, such as piperidine, piperazine, aziridine, pyrrolidine, morpholine, and their derivatives.

[0056] As used in this disclosure, the term "salt" refers to an acidic and / or basic salt formed with inorganic and / or organic acids and bases.

[0057] As used herein, the term "pharmaceutically acceptable salt" means that salt, within reasonable medical judgment, is suitable for contact with mammalian tissues without excessive toxicity, irritation, anaphylactic reactions, etc., and is proportionate to a reasonable benefit / risk ratio. Pharmaceutically acceptable salts are well known in the art. For example, SM Berge et al. described pharmaceutically acceptable salts in detail in J. PharmaceuticalSciences, 1977, 66, 1–19, and their related teachings are incorporated herein by reference in their entirety. Pharmaceutically acceptable salts of compounds and lipids described herein include pharmaceutically acceptable acid addition salts and pharmaceutically acceptable base addition salts. In some embodiments, one or more components of a pharmaceutically acceptable salt are not acid addition salts or base addition salts. The term "pharmaceutically acceptable salt" is intended to include salts derived from inorganic or organic acids, including, for example, hydrochloric acid, hydrobromic acid, sulfuric acid, nitric acid, perchloric acid, phosphoric acid, formic acid, acetic acid, lactic acid, maleic acid, fumaric acid, succinic acid, tartaric acid, glycolic acid, salicylic acid, citric acid, methanesulfonic acid, benzenesulfonic acid, benzoic acid, malonic acid, trifluoroacetic acid, trichloroacetic acid, naphthalene-2-sulfonic acid, and other acids. Pharmaceutically acceptable salt forms may also include those in which the ratio of the molecules constituting the salt is not 1:1. For example, the salt may contain more than one molecule of inorganic or organic acid per molecule of base, such as the formula provided herein, where each molecule corresponds to two molecules of hydrochloric acid. As another example, the salt may contain fewer than one molecule of inorganic or organic acid per molecule of base, such as compounds or lipids of the formula provided herein, where each molecule of tartaric acid corresponds to two molecules. Other examples of pharmaceutically acceptable acid addition salts are salts formed by amino groups with inorganic acids (e.g., hydrochloric acid, hydrobromic acid, phosphoric acid, sulfuric acid, and perchloric acid) or with organic acids (e.g., acetic acid, oxalic acid, maleic acid, tartaric acid, citric acid, succinic acid, or malonic acid) or by using other methods used in the art (e.g., ion exchange).Other pharmaceutically acceptable acid addition salts include adipate, alginate, ascorbate, aspartate, benzenesulfonate, benzoate, hydrogen sulfate, borate, butyrate, camphorate, camphorsulfonate, cinnamate, citrate, cyclopentanepropionate, disaccharide, dodecyl sulfate, ethanesulfonate, formate, fumarate, glucono-p-ethylhexanoate, glycerophosphate, gluconate, glutarate, glycolate, hemisulfate, heptanate, hexanoate, hydroiodide, hydroxybenzoate, 2-hydroxy-ethanesulfonate, hydroxymaleate, and lactobionate. Lactates, laurates, lauryl sulfates, malates, maleates, malonates, methanesulfonates, 2-naphthalenesulfonates, nicotinates, nitrates, oleates, oxalates, palmitates, papoate, pectinates, persulfates, 2-phenoxybenzoates, phenylacetates, 3-phenylpropionates, phosphates, p-pentanoates, propionates, pyruvates, salicylates, stearates, succinates, sulfates, tartrates, thiocyanates, p-toluenesulfonates, undecanoates, valerates, etc. These salts can form monoacids, diacids, or triacids, and can exist in hydrated, solvated, or essentially anhydrous forms.

[0058] Pharmaceutically acceptable base addition salts include those formed with inorganic bases such as alkali metals, alkaline earth metals, and ammonium bases, as well as those formed with aliphatic, alicyclic, or aromatic organic amines such as methylamine, trimethylamine, and methylpyridine, or N⁺((C1-C4)alkyl)⁴ salts. Representative alkali metal or alkaline earth metal salts include sodium, lithium, potassium, calcium, magnesium, and barium. Other pharmaceutically acceptable base addition salts, where appropriate, include those formed using counterions such as halides, hydroxides, carboxyl groups, sulfates, phosphates, nitrates, lower alkyl sulfonates, and aryl sulfonates with non-toxic ammonium, quaternary ammonium, and amine cations.

[0059] As used herein and as is well known in the art, “treatment” is a method for achieving a beneficial or desired outcome, including clinical outcomes. Beneficial or desired clinical outcomes may include, but are not limited to, the reduction or improvement of one or more symptoms or conditions, a reduction in the severity of the disease, stabilization (i.e., non-deterioration) of the disease state, prevention of disease spread, delay or slowing of disease progression, improvement or mitigation of the disease state, and remission (whether partial or complete), whether detectable or undetectable. “Treatment” may also refer to prolonged survival compared to expected survival without treatment.

[0060] In some embodiments, as used herein, “treatment” refers to taking steps to deliver a therapeutic agent to an individual (e.g., a mammal) in need (e.g., by administering one or more therapeutic agents to the individual). In some embodiments, “treatment” also includes suppressing a disease or symptom (e.g., by slowing or stopping its progression or causing the disease or symptom to subside) and alleviating the symptoms caused by the disease or symptom.

[0061] As used in this article, “individual” includes humans, livestock such as laboratory animals (e.g., dogs, monkeys, pigs, rats, mice, etc.), pets (e.g., cats, dogs, rabbits, etc.), and animals (e.g., pigs, cattle, sheep, goats, horses, etc.), as well as non-livestock. In some respects, an individual is a human being.

[0062] As used herein, an “effective amount” or “therapeutic effective amount” of a composition is the amount of the composition that produces a identifiable physiological effect in an individual or animal when administered to an individual. When administered at a therapeutic effective amount, the compositions disclosed herein may have pharmacological properties to provide physiological effects for the treatment of a variety of physiological conditions. The actual amount including an “effective amount” or “therapeutic effective amount” will vary depending on a variety of conditions, including but not limited to the dosage, the specific condition being treated, the severity of the condition, the patient’s body size and health status, and the route of administration. A skilled medical practitioner can readily determine the appropriate amount using methods known in the medical field.

[0063] Furthermore, "therapeutic effective amount" can refer to the amount that effectively achieves the desired therapeutic outcome (e.g., treatment, cure, suppression, or improvement of physiological responses or conditions) within the necessary dosage and time period. A complete therapeutic effect does not necessarily occur with the administration of a single dose and can occur only after a series of doses. Therefore, the therapeutic effective amount can be administered in one or more doses. Therapeutic effective amount can vary depending on factors such as the mammal's disease state, age, sex, and weight, the administration method, and the ability of the therapeutic agent or combination of therapeutic agents to elicit the desired response in the individual. Therapeutic effective amount of the agent to be administered can be determined by those skilled in the art using the guidance provided herein and other methods known in the art.

[0064] B. Example Implementation Plan Exemplary implementations of this disclosure include: 1. A composition comprising a plurality of lipid nanoparticles, wherein the lipid nanoparticles comprise a pharmaceutical agent, an ionizable lipid, and a fully saturated cationic lipid, wherein the lipid nanoparticles are free of zwitterionic phospholipids.

[0065] 2. The composition according to embodiment 1, wherein the lipid nanoparticles have an apparent ionization constant (pKa) of about 4.5 to about 7.

[0066] 3. The composition according to embodiment 1 or 2, wherein the lipid nanoparticles have an apparent ionization constant (pKa) of about 5 to about 7.

[0067] 4. The composition according to any one of embodiments 1-3, wherein the lipid nanoparticles have an apparent ionization constant (pKa) of about 5.5 to about 7.

[0068] 5. The composition according to any one of embodiments 1-4, wherein the lipid nanoparticles have an apparent ionization constant (pKa) of about 6 to about 7.

[0069] 6. The composition according to any one of embodiments 1-5, wherein the cationic lipid is less than about 50 mol of the lipid nanoparticles.

[0070] 7. The composition according to any one of embodiments 1-6, wherein the cationic lipid is less than about 30 mol of the lipid nanoparticles.

[0071] 8. The composition according to any one of embodiments 1-7, wherein the cationic lipid is less than about 20 mol of the lipid nanoparticles.

[0072] 9. The composition according to any one of embodiments 1-8, wherein the fully saturated cationic lipid is C 12 -C 18 Fully saturated cationic lipids.

[0073] 10. The composition according to any one of embodiments 1-9, further comprising sterols.

[0074] 11. The composition according to any one of embodiments 1-10, further comprising PEG-lipids.

[0075] 12. The composition according to any one of embodiments 1-11, wherein the pharmaceutical agent is a bioactive agent.

[0076] 13. The composition according to embodiment 12, wherein the bioactive agent is a therapeutic agent.

[0077] 14. The composition according to embodiment 13, wherein the therapeutic agent is a nutritional supplement.

[0078] 15. The composition according to any one of embodiments 1-11, wherein the agent is a developer.

[0079] 16. The composition according to any one of embodiments 1-11, wherein the pharmaceutical agent is a diagnostic agent.

[0080] 17. The composition according to any one of embodiments 1-11, wherein the agent is a nucleic acid.

[0081] 18. The composition according to embodiment 17, wherein the nucleic acid is ribonucleic acid.

[0082] 19. The composition according to embodiment 17, wherein the nucleic acid is deoxyribonucleic acid.

[0083] 20. The composition according to embodiment 17, wherein the nucleic acid is a non-natural nucleic acid.

[0084] 21. The composition according to any one of embodiments 1-20, wherein the ionizable lipid is a lipid of formula I or II or a pharmaceutically acceptable salt thereof: in X is -NR 1 -or -O-; R 1 It is a hydrogen atom, a hydrocarbon group with 6 to 24 atoms, or R 21 -L 1 -R 22 -; R 21 It is a hydrocarbon group having 1 to 24 carbon atoms; L 1 Is -O(CO)O-, -O(CO)-, -(CO)O-, -O- or ; R 22 It is a divalent hydrocarbon linking group having 1 to 18 carbon atoms; R 2 and R 3 Each is independently a hydrogen atom, a hydrocarbon group having 3 to 24 carbon atoms, or an R... 31 -L 1 -R 32 -; R 31 It is a hydrocarbon group having 1 to 24 carbon atoms; L 2 Is -O(CO)O-, -O(CO)-, -(CO)O-, -O- or ; R 32 It is a divalent hydrocarbon linking group having 1 to 18 carbon atoms; R 4 R 5 R 6 R 7 R 8 R 9 R10 R 11 and R 12 Each is independently a hydrogen atom or optionally R 33 Substituted alkyl groups having 1 to 18 carbon atoms, wherein the alkyl group is selected from R 4 and R 5 R 10 and R 5 R 5 and R 12 R 4 and R 6 R 5 and R 6 R 6 and R 7 R 6 and R 10 R 12 and R 7 and R 7 and R 8 One or more pairs of groups can be linked together to form a 4- to 7-membered ring of carbon and nitrogen atoms, said ring may contain oxygen atoms; R 33 It is a hydroxyl group, a carboxyl group, or -NR. 45 R 46 -O(CO)OR 41 -O(CO)-R 42 -(CO)OR 43 -OR 44 Or optionally R 34 Substituted aryl or heteroaryl groups; R 34 It is an alkyl group, hydroxy group, carboxyl group, or -NR group having 1 to 18 carbon atoms. 45 R 46 -O(CO)OR 41 -O(CO)-R 42 -(CO)OR 43 or -OR 44 ; R 41 R 42 R 43 R 44 R 45 and R 46 Each is independently a hydrocarbon group having 1 to 18 carbon atoms; a, b, c, and d are each independent integers from 0 to 3, where a+b is 1 or greater, and c+d is 1 or greater; R 51 and R 52 Each independently is arbitrarily R 35Substituted alkyl groups having 1 to 18 carbon atoms; R 35 It is a hydroxyl group, -G 20 -CH(R 55 (R) 56 ), -N(R 58 (R) 59 ) or -G 20 -R 60 ; G 20 It is -(CO)O- or -O(CO)-; R 55 and R 56 Each is independently hydrogen or an alkyl group having 1 to 18 carbon atoms; R 58 and R 59 Each independently is hydrogen or optionally R 36 Substituted cycloalkyl groups having 3 to 6 carbon atoms; R 60 It is an alkyl group having 1 to 18 carbon atoms or a cycloalkyl group having 3 to 6 carbon atoms; R 36 It is -N(R) 61 (R) 62 ) or -G 20 -R 65 ; R 61 and R 62 Each is independently hydrogen or a cycloalkyl group having 3 to 6 carbon atoms; R 65 It is an alkyl group having 1 to 18 carbon atoms, -L 40 -CH(R 66 (R) 67 ) or -G 20 -R 66 ; L 40 It is a divalent alkyl group containing 1 to 6 carbon atoms; R 66 and R 67 Each is independently an alkyl group containing 1 to 10 carbon atoms or an alkoxy group containing 1 to 10 carbon atoms; L 10 It is a divalent alkyl group containing 1 to 10 carbon atoms; G 30 -S(CO)N(R) 64 )-; R 64 Yes -L 30 -G 20 -CH(R55 (R) 56 ); a' is 0 or 1; G 10 It is G 20 -O(CO)O- or -N(R) 63 )C(O)-; R 63 It is an alkyl group containing 1 to 18 carbon atoms; L 20 It is a divalent alkyl group containing 1 to 6 carbon atoms; b' is 0 or 1; R 53 R 54 and R 57 Each independently is hydrogen or optionally R 36 The substituted alkyl group containing 1 to 18 carbon atoms; and L 30 It is a single bond or an alkyl group containing 1 to 18 carbon atoms.

[0085] 22. The composition according to any one of embodiments 1-20, wherein the ionizable lipid is a lipid according to formula VII or a pharmaceutically acceptable salt thereof: in R 1 and R 2 Each is independently a hydrocarbon group having 1 to 18 carbon atoms, and R 3 It is a hydrocarbon group having 2 to 8 carbon atoms, wherein R 1 R 2 and R 3 The hydrocarbon groups represented are each independently and optionally surrounded by one or more groups selected from -OH, COOH, -NR. 51 R 52 -OC(O)OR 53 -C(O)OR 54 -OC(O)-R 55 and -OR 56 Substituents of the substituents; R 4 It is a hydrocarbon group having 1 to 8 carbon atoms; R 5 and R 6 Each is independently a hydrocarbon group having 1 to 8 carbon atoms, or -R 8 -L 1 -R 9 Exclude R 5 and R 6 Both cases involve hydrocarbon groups having 1 to 8 carbon atoms; R 7 Yes -R 10 -L 2 -R 11 -L 3 -R 12 ; R 51 and R 52 Each is an independent hydrocarbon group having 1 to 8 carbon atoms; R 53 R 54 R 55 and R 56 Each is an independent hydrocarbon group having 1 to 24 carbon atoms; By R 53 R 54 R 55 and R 56 Each of the hydrocarbon groups represented is independently and optionally surrounded by an aryl group having 6 to 20 carbon atoms or -SR 58 The aryl group having 6 to 20 carbon atoms is optionally replaced by -OH, COOH, or -NR. 51 R 52 -OC(O)OR 53 -C(O)OR 54 -OC(O)-R 55 -OR 56 or -R 59 -R 57 Replace; among them R 58 and R 59 Each is an independent hydrocarbon group having 1 to 12 carbon atoms; R 57 -OH, COOH, -NR 61 R 62 -OC(O)OR 63 -C(O)OR 64 -OC(O)-R 65 or -OR 66 ; R 61 and R 62 Each is an independent hydrocarbon group having 1 to 8 carbon atoms; R 63 R 64 R 65 and R 66 Each is an independent hydrocarbon group having 1 to 24 carbon atoms; By R 63 R 64 R 65 and R 66Each of the hydrocarbon groups represented is independently and optionally surrounded by an aryl group having 6 to 20 carbon atoms or -SR 68 The aryl group having 6 to 20 carbon atoms can be replaced by -OH, COOH, -NR, etc. 61 R 62 -OC(O)OR 63 -C(O)OR 64 -OC(O)-R 65 -OR 66 or -R 69 -R 67 replace; R 68 and R 69 Each is an independent hydrocarbon group having 1 to 12 carbon atoms; R 67 -OH, COOH, -NR 61 R 62 -OC(O)OR 63 -C(O)OR 64 -OC(O)-R 65 or -OR 66 L 1 L 2 and L 3 Each can be independently represented as -OC(O)O-, -C(O)O-, -OC(O)-, or -O-; R 8 It is a hydrocarbon group having 1 to 12 carbon atoms; R 9 It is a hydrocarbon group having 1 to 24 carbon atoms; R 10 It is a hydrocarbon group having 1 to 8 carbon atoms; R 11 It is a hydrocarbon group having 1 to 24 carbon atoms; R 12 It is a hydrocarbon group having 1 to 24 carbon atoms; By R 9 and R 12 The hydrocarbon groups represented are each independently and optionally replaced by aryl groups, -OC(O)OR 53 -C(O)OR 54 -OC(O)-R 55 or -SR 58 Replace; and By R 11 The hydrocarbon groups represented are each independently and optionally labeled -OC(O)OR 53 -C(O)OR 54 or -OC(O)-R 55 replace.

[0086] 23. The composition according to any one of embodiments 1-21, wherein the ionizable lipid is a lipid of formula III or a pharmaceutically acceptable salt thereof: in R 2 and R 3 Each is independently a hydrogen atom, a hydrocarbon group having 3 to 24 carbon atoms, or an R... 31 -L 2 -R 32 -; R 31 It is a hydrocarbon group having 1 to 24 carbon atoms; L 2 Is -O(CO)O-, -O(CO)-, -(CO)O-, -O- or ; R 32 It is a divalent hydrocarbon linking group having 1 to 18 carbon atoms; R 5 It is a hydrogen atom or optionally R 33 Substituted alkyl groups having 1 to 18 carbon atoms; R 7 and R 8 Each is independently a hydrogen atom or optionally R 33 Substituted alkyl groups having 1 to 18 carbon atoms; R 33 It is a hydroxyl group, a carboxyl group, or -NR. 45 R 46 -O(CO)OR 41 -O(CO)-R 42 -(CO)OR 43 -OR 44 Or optionally R 34 Substituted aryl or heteroaryl groups; R 34 It is an alkyl group, hydroxy group, carboxyl group, or -NR group having 1 to 18 carbon atoms. 45 R 46 -O(CO)OR 41 -O(CO)-R 42 -(CO)OR 43 or -OR 44 ; R 41 R 42 R 43 R 44 R 45 and R46 Each is independently a hydrocarbon group having 1 to 18 carbon atoms; and e is an integer selected from 2 or 3.

[0087] 24. The composition according to any one of embodiments 1-23, wherein the ionizable lipid is selected from: and .

[0088] 25. The composition according to any one of embodiments 1-20 and 22, wherein the ionizable lipid is selected from: and .

[0089] 26. The composition according to any one of embodiments 1-25, wherein the cationic lipid is a lipid according to formula IV: in: R 101 and R 102 Each is independently an optional substitution (C8-C) 24 )alkyl or optionally substituted (C8-C 24 alkenyl; R 103 In each case, it is independently an optionally substituted (C1-C6) alkyl group; R 104 It is an optionally substituted (C1-C6) alkyl group; and X⁻ is a monovalent anion.

[0090] 27. The composition according to any one of embodiments 1-26, wherein the cationic lipid is 1,2-dilauroyl- sn -Glyceryl-O-ethyl-3-phosphocholine (EDLPC), 1,2-dimyristoyl- sn -Glyceryl-O-ethyl-3-phosphocholine (EDMPC), 1,2-dimyristoleoyl- sn -Glyceryl-O-ethyl-3-phosphocholine (14:1 EPC), 1,2-dipalmitoyl- sn -Glyceryl-O-ethyl-3-phosphocholine (EDPPC), 1,2-distearate- sn -Glyceryl-O-ethyl-3-phosphocholine (EDSPC), 1,2-dioleoyl- sn-Glyceryl-3-ethylphosphorylcholine (EDOPC) or 1-palmitoyl-2-oleoyl- sn -Glyceryl-O-ethyl-3-phosphocholine (16:0-18:1 EPC).

[0091] 28. The composition according to any one of embodiments 1-27, wherein the cationic lipid is EDPPC.

[0092] 29. The composition according to any one of embodiments 1-25, wherein the cationic lipid is a lipid according to formula V: in: R 101 and R 102 Each is independently an optional substitution (C8-C) 24 )alkyl or optionally substituted (C8-C 24 alkenyl; R 103 In each case, it is independently an optionally substituted (C1-C6) alkyl group; and X⁻ is a monovalent anion.

[0093] 30. The composition according to any one of embodiments 1-25 and 29, wherein the cationic lipid is selected from 1,2-dimyristoyl-3-trimethylammonium-propane (DMTAP), 1,2-dipalmitoyl-3-trimethylammonium-propane (DPTAP), 1,2-distearate-3-trimethylammonium-propane (DSTAP), 1,2-dioleoyl-3-trimethylammonium-propane (DOTAP) and N-(2-hydroxyethyl)-N,N-dimethyl-2,3-bis(oleoyloxy)prop-1-ammonium (DORI).

[0094] 31. The composition according to any one of embodiments 1-25, 29 and 30, wherein the cationic lipid is DPTAP.

[0095] 32. The composition according to any one of embodiments 1-25, wherein the cationic lipid is a lipid according to formula VI: in: R 101 and R 102 Each is independently an optional substitution (C8-C) 24 )alkyl or optionally substituted (C8-C 24 alkenyl; R 105 Each is independently a (C1-C6) alkyl group; and X⁻ is a monovalent anion.

[0096] 33. The composition according to any one of embodiments 1-25 and 32, wherein the cationic lipid is .

[0097] 34. A composition comprising a plurality of lipid nanoparticles, wherein the lipid nanoparticles comprise a pharmaceutical agent, an ionizable lipid, and a cationic lipid, wherein the cationic lipid is a lipid according to formula VI: in: R 101 and R 102 Each is independently an optional substitution (C8-C) 24 )alkyl or optionally substituted (C8-C 24 alkenyl; R 105 Each is independently a (C1-C6) alkyl group; and X⁻ is a monovalent anion.

[0098] 35. The composition according to embodiment 34, wherein the cationic lipid is .

[0099] 36. A composition comprising a plurality of lipid nanoparticles, wherein the lipid nanoparticles comprise a pharmaceutical agent, an ionizable lipid, and a cationic lipid, wherein the ionizable lipid is a lipid of formula I or a pharmaceutically acceptable salt thereof: in X is -NR 1 -or -O-; R 1 It is a hydrogen atom, a hydrocarbon group with 6 to 24 atoms, or R 21 -L 1 -R 22 -; R 21 It is a hydrocarbon group having 1 to 24 carbon atoms; L 1 Is -O(CO)O-, -O(CO)-, -(CO)O-, -O- or ; R 22 It is a divalent hydrocarbon linking group having 1 to 18 carbon atoms; R 2 and R 3 Each is independently a hydrogen atom, a hydrocarbon group having 3 to 24 carbon atoms, or an R... 31 -L 1-R 32 -; R 31 It is a hydrocarbon group having 1 to 24 carbon atoms; L 2 Is -O(CO)O-, -O(CO)-, -(CO)O-, -O- or ; R 32 It is a divalent hydrocarbon linking group having 1 to 18 carbon atoms; R 4 R 5 R 6 R 7 R 8 R 9 R 10 R 11 and R 12 Each is independently a hydrogen atom or optionally R 33 Substituted alkyl groups having 1 to 18 carbon atoms, wherein the alkyl group is selected from R 4 and R 5 R 10 and R 5 R 5 and R 12 R 4 and R 6 R 5 and R 6 R 6 and R 7 R 6 and R 10 R 12 and R 7 and R 7 and R 8 One or more pairs of groups can be linked together to form a 4- to 7-membered ring of carbon and nitrogen atoms, said ring may contain oxygen atoms; R 33 It is a hydroxyl group, a carboxyl group, or -NR. 45 R 46 -O(CO)OR 41 -O(CO)-R 42 -(CO)OR 43 -OR 44 Or optionally R 34 Substituted aryl or heteroaryl groups; R 34 It is an alkyl group, hydroxy group, carboxyl group, or -NR group having 1 to 18 carbon atoms. 45 R 46 -O(CO)OR 41 -O(CO)-R 42-(CO)OR 43 or -OR 44 ; R 41 R 42 R 43 R 44 R 45 and R 46 Each is independently a hydrocarbon group having 1 to 18 carbon atoms; and a, b, c, and d are each independent integers from 0 to 3, where a+b is 1 or greater, and c+d is 1 or greater.

[0100] 37. The composition according to embodiment 36, wherein the ionizable lipid is a lipid of formula III or a pharmaceutically acceptable salt thereof: in R 2 and R 3 Each is independently a hydrogen atom, a hydrocarbon group having 3 to 24 carbon atoms, or an R... 31 -L 2 -R 32 -; R 31 It is a hydrocarbon group having 1 to 24 carbon atoms; L 2 Is -O(CO)O-, -O(CO)-, -(CO)O-, -O- or ; R 32 It is a divalent hydrocarbon linking group having 1 to 18 carbon atoms; R 5 It is a hydrogen atom or optionally R 33 Substituted alkyl groups having 1 to 18 carbon atoms; R 33 It is a hydroxyl group, a carboxyl group, or -NR. 45 R 46 -O(CO)OR 41 -O(CO)-R 42 -(CO)OR 43 -OR 44 Or optionally R 34 Substituted aryl or heteroaryl groups; R 7 and R 8 Each is independently a hydrogen atom or optionally R 33 Substituted alkyl groups having 1 to 18 carbon atoms; R 34It is an alkyl group, hydroxy group, carboxyl group, or -NR group having 1 to 18 carbon atoms. 45 R 46 -O(CO)OR 41 -O(CO)-R 42 -(CO)OR 43 or -OR 44 ; R 41 R 42 R 43 R 44 R 45 and R 46 Each is independently a hydrocarbon group having 1 to 18 carbon atoms; and e is an integer selected from 2 or 3.

[0101] 38. The composition according to embodiment 36 or 37, wherein the ionizable lipid is selected from: and .

[0102] 39. A composition comprising a plurality of lipid nanoparticles, wherein the lipid nanoparticles comprise a pharmaceutical agent, an ionizable lipid, and a cationic lipid, wherein the ionizable lipid is a lipid according to Formula VII or a pharmaceutically acceptable salt thereof: in R 1 and R 2 Each is independently a hydrocarbon group having 1 to 18 carbon atoms, and R 3 It is a hydrocarbon group having 2 to 8 carbon atoms, wherein R 1 R 2 and R 3 The hydrocarbon groups represented are each independently and optionally surrounded by one or more groups selected from -OH, COOH, -NR. 51 R 52 -OC(O)OR 53 -C(O)OR 54 -OC(O)-R 55 and -OR 56 Substituents of the substituents; R 4 It is a hydrocarbon group having 1 to 8 carbon atoms; R 5 and R 6 Each is independently a hydrocarbon group having 1 to 18 carbon atoms, or -R 8 -L 1-R 9 Exclude R 5 and R 6 Both cases involve hydrocarbon groups having 1 to 8 carbon atoms; R 7 Yes -R 10 -L 2 -R 11 -L 3 -R 12 ; R 51 and R 52 Each is an independent hydrocarbon group having 1 to 8 carbon atoms; R 14 R 53 R 54 R 55 and R 56 Each is an independent hydrocarbon group having 1 to 24 carbon atoms; By R 53 R 54 R 55 and R 56 Each of the hydrocarbon groups represented is independently and optionally surrounded by an aryl group having 6 to 20 carbon atoms or -SR 58 The aryl group having 6 to 20 carbon atoms is optionally replaced by -OH, COOH, or -NR. 51 R 52 -OC(O)OR 53 -C(O)OR 54 -OC(O)-R 55 -OR 56 or -R 59 -R 57 Replace; where R 58 and R 59 Each is independently a hydrocarbon group having 1 to 12 carbon atoms; R 57 -OH, COOH, -NR 61 R 62 -OC(O)OR 63 -C(O)OR 64 -OC(O)-R 65 or -OR 66 ; R 61 and R 62 Each is an independent hydrocarbon group having 1 to 8 carbon atoms; R 63 R 64 R 65 and R 66Each is an independent hydrocarbon group having 1 to 24 carbon atoms; By R 63 R 64 R 65 and R 66 Each of the hydrocarbon groups represented is independently and optionally surrounded by an aryl group having 6 to 20 carbon atoms or -SR 68 The aryl group having 6 to 20 carbon atoms can be replaced by -OH, COOH, -NR, etc. 61 R 62 -OC(O)OR 63 -C(O)OR 64 -OC(O)-R 65 -OR 66 or -R 69 -R 67 replace; R 68 and R 69 Each is an independent hydrocarbon group having 1 to 12 carbon atoms; R 67 -OH, COOH, -NR 61 R 62 -OC(O)OR 63 -C(O)OR 64 -OC(O)-R 65 or -OR 66 ; L 1 L 2 and L 3 Each can be independently -OC(O)O-, -C(O)O-, -OC(O)-, or -O-; R 8 It is a hydrocarbon group having 1 to 12 carbon atoms; R 9 It is a hydrocarbon group having 1 to 24 carbon atoms; R 10 It is a hydrocarbon group having 1 to 8 carbon atoms; R 11 It is a hydrocarbon group having 1 to 24 carbon atoms; R 12 It is a hydrocarbon group having 1 to 24 carbon atoms; By R 9 and R 12 The hydrocarbon groups represented are each independently and optionally replaced by aryl groups, -OC(O)OR 53 -C(O)OR 54 -OC(O)-R 55 or -SR 58 Replace; and By R11 and R 14 The hydrocarbon groups represented are each independently and optionally labeled -OC(O)OR 53 -C(O)OR 54 or -OC(O)-R 55 replace.

[0103] 40. The composition according to embodiment 39, wherein the ionizable lipid is selected from: and .

[0104] 41. A composition comprising a plurality of lipid nanoparticles, wherein the lipid nanoparticles comprise a pharmaceutical agent, an ionizable lipid, and at least one of EDPPC and DPTAP, wherein the ionizable lipid is a lipid of formula I or II or a pharmaceutically acceptable salt thereof: in X is -NR 1 -or -O-; R 1 It is a hydrogen atom, a hydrocarbon group with 6 to 24 atoms, or R 21 -L 1 -R 22 -; R 21 It is a hydrocarbon group having 1 to 24 carbon atoms; L 1 Is -O(CO)O-, -O(CO)-, -(CO)O-, -O- or ; R 22 It is a divalent hydrocarbon linking group having 1 to 18 carbon atoms; R 2 and R 3 Each is independently a hydrogen atom, a hydrocarbon group having 3 to 24 carbon atoms, or an R... 31 -L 1 -R 32 -; R 31 It is a hydrocarbon group having 1 to 24 carbon atoms; L 2 Is -O(CO)O-, -O(CO)-, -(CO)O-, -O- or ; R 32 It is a divalent hydrocarbon linking group having 1 to 18 carbon atoms; R 4 R5 R 6 R 7 R 8 R 9 R 10 R 11 and R 12 Each is independently a hydrogen atom or optionally R 33 Substituted alkyl groups having 1 to 18 carbon atoms, wherein the alkyl group is selected from R 4 and R 5 R 10 and R 5 R 5 and R 12 R 4 and R 6 R 5 and R 6 R 6 and R 7 R 6 and R 10 R 12 and R 7 and R 7 and R 8 One or more pairs of groups can be linked together to form a 4- to 7-membered ring of carbon and nitrogen atoms, said ring may contain oxygen atoms; R 33 It is a hydroxyl group, a carboxyl group, or -NR. 45 R 46 -O(CO)OR 41 -O(CO)-R 42 -(CO)OR 43 -OR 44 Or optionally R 34 Substituted aryl or heteroaryl groups; R 34 It is an alkyl group, hydroxy group, carboxyl group, or -NR group having 1 to 18 carbon atoms. 45 R 46 -O(CO)OR 41 -O(CO)-R 42 -(CO)OR 43 or -OR 44 ; R 41 R 42 R 43 R 44 R 45 and R 46 Each is independently a hydrocarbon group having 1 to 18 carbon atoms; a, b, c, and d are each independent integers from 0 to 3, where a+b is 1 or greater, and c+d is 1 or greater; R 51 and R 52 Each independently is arbitrarily R 35 Substituted alkyl groups having 1 to 18 carbon atoms; R 35 It is a hydroxyl group, -G 20 -CH(R 55 (R) 56 ), -N(R 58 (R) 59 ) or -G 20 -R 60 ; G 20 It is -(CO)O- or -O(CO)-; R 55 and R 56 Each is independently hydrogen or an alkyl group having 1 to 18 carbon atoms; R 58 and R 59 Each independently is hydrogen or optionally R 36 Substituted cycloalkyl groups having 3 to 6 carbon atoms; R 60 It is an alkyl group having 1 to 18 carbon atoms or a cycloalkyl group having 3 to 6 carbon atoms; R 36 It is -N(R) 61 (R) 62 ) or -G 20 -R 65 ; R 61 and R 62 Each is independently hydrogen or a cycloalkyl group having 3 to 6 carbon atoms; R 65 It is an alkyl group having 1 to 18 carbon atoms, -L 40 -CH(R 66 (R) 67 ) or -G 20 -R 66 ; L 40 It is a divalent alkyl group containing 1 to 6 carbon atoms; R 66 and R 67 Each is independently an alkyl group containing 1 to 10 carbon atoms or an alkoxy group containing 1 to 10 carbon atoms; L 10 It is a divalent alkyl group containing 1 to 10 carbon atoms; G 30 -S(CO)N(R) 64 )-; R 64 Yes -L 30 -G 20 -CH(R 55 (R) 56 ); a' is 0 or 1; G 10 It is G 20 -O(CO)O- or -N(R) 63 )C(O)-; R 63 It is an alkyl group containing 1 to 18 carbon atoms; L 20 It is a divalent alkyl group containing 1 to 6 carbon atoms; b' is 0 or 1; R 53 R 54 and R 57 Each independently is hydrogen or optionally R 36 The substituted alkyl group containing 1 to 18 carbon atoms; and L 30 It is a single bond or an alkyl group containing 1 to 18 carbon atoms.

[0105] 42. The composition according to any one of embodiments 34-41, wherein the pharmaceutical agent is a bioactive agent.

[0106] 43. The composition according to embodiment 42, wherein the bioactive agent is a therapeutic agent.

[0107] 44. The composition according to embodiment 43, wherein the therapeutic agent is a nutritional supplement.

[0108] 45. The composition according to any one of embodiments 34-41, wherein the agent is a developer.

[0109] 46. ​​The composition according to any one of embodiments 34-41, wherein the pharmaceutical agent is a diagnostic agent.

[0110] 47. The composition according to any one of embodiments 34-41, wherein the agent is a nucleic acid.

[0111] 48. The composition according to embodiment 47, wherein the nucleic acid is ribonucleic acid.

[0112] 49. The composition according to embodiment 47, wherein the nucleic acid is deoxyribonucleic acid.

[0113] 50. The composition according to embodiment 47, wherein the nucleic acid is a non-natural nucleic acid.

[0114] 51. A method of delivering a drug to one or more of the brain, heart, muscles, and kidneys of an individual in need, the method comprising systemically administering to the individual a composition comprising a plurality of lipid nanoparticles, wherein the lipid nanoparticles comprise the drug, ionizable lipids, and fully saturated cationic lipids.

[0115] 52. The method according to embodiment 51, wherein the agent is delivered to the brain.

[0116] 53. The method according to embodiment 51, wherein the agent is delivered to the heart.

[0117] 54. The method according to embodiment 51, wherein the agent is delivered to muscle.

[0118] 55. The method according to embodiment 51, wherein the agent is delivered to the kidney.

[0119] 56. The method according to any one of embodiments 51-55, wherein the ionizable lipid is a lipid of formula (I).

[0120] 57. The method according to any one of embodiments 51-56, wherein the ionizable lipid is a lipid of formula (III).

[0121] 58. The method according to any one of embodiments 51-55, wherein the ionizable lipid is a lipid of formula (II).

[0122] 59. The method according to any one of embodiments 51-55, wherein the ionizable lipid is a lipid of formula (VII).

[0123] 60. The method according to any one of embodiments 51-59, wherein the cationic lipid is a lipid of formula (IV).

[0124] 61. The method according to any one of embodiments 51-59, wherein the cationic lipid is a lipid of formula (V).

[0125] 62. The method according to any one of embodiments 51-59, wherein the cationic lipid is a lipid of formula (VI).

[0126] 63. A method for delivering a pharmaceutical agent to an individual in need, the method comprising administering to the individual the composition described in any one of embodiments 1-33.

[0127] 64. A method for delivering a pharmaceutical agent to an individual in need, the method comprising administering to the individual the composition described in any one of embodiments 34-35.

[0128] 65. A method for delivering a pharmaceutical agent to an individual in need, the method comprising administering to the individual the composition described in any one of embodiments 36-38.

[0129] 66. A method for delivering a pharmaceutical agent to an individual in need, the method comprising administering to the individual the composition described in any one of embodiments 39-50.

[0130] 67. A method for treating an individual suffering from a disease, disorder, or ailment that is beneficially treatable by a pharmaceutical agent, the method comprising administering to the individual a therapeutically effective amount of the composition of any one of embodiments 1-50.

[0131] C. Lipid particles In some embodiments, the lipid particles of this disclosure comprise ionizable lipids, cationic lipids, and a pharmaceutical agent. In some embodiments, the lipid particles further comprise sterols. In some embodiments, the lipid particles further comprise phospholipids. In some embodiments, the lipid particles further comprise PEG-lipids. Exemplary and preferred lipids of each type are provided herein. In some embodiments, the molar percentage distribution of the ionizable lipids and cationic lipids is 10-90:10-90, wherein the sum of each molar percentage must be 100. In some embodiments, the molar percentage distribution of the ionizable lipids, cationic lipids, and sterols is 10-90:10-90:10-90, wherein the sum of each molar percentage must be 100. In some embodiments, the molar percentage distribution of the ionizable lipids, cationic lipids, and sterols is 15-75:5-75:25-80, wherein the sum of each molar percentage must be 100. In some embodiments, the molar percentage distribution of the ionizable lipids, cationic lipids, and sterols is 20-60:15-50:20-60, wherein the sum of each molar percentage must be 100. In some embodiments, the molar percentage distribution of the ionizable lipids, cationic lipids, sterols, and phospholipids is 15-75:5-75:25-80:0.1-10, wherein the sum of each molar percentage must be 100. In some embodiments, the molar percentage distribution of the ionizable lipids, cationic lipids, sterols, and phospholipids is 20-60:15-50:20-60:2.5-7.5, wherein the sum of each molar percentage must be 100. In some embodiments, the molar percentage distribution of the ionizable lipids, cationic lipids, sterols, phospholipids, and PEG lipids is 20-60:15-50:20-60:2.5-7.5:0.5-2.5, wherein the sum of each molar percentage must be 100. In some embodiments, the molar percentage distribution of the ionizable lipids, cationic lipids, sterols and PEG lipids is 20-60:15-50:20-60:0.5-2.5, wherein the sum of each molar percentage must be 100.

[0132] As used herein, a "lipid particle" refers to a particle composed of at least one lipid. Lipid particles can have various structures; for example, according to this disclosure, lipid particles can take the form of lipid aggregates, micelles, liposomes, cationic liposome complexes, lipid nanoparticles, etc. For example, in some embodiments, the lipid particle has a single lipid bilayer and an internal aqueous phase. In other embodiments, the lipid particle is a liposome having multiple lipid bilayers stacked together with multiple internal aqueous phase compartments. The bilayer of the lipid particles of this disclosure can be substantially gel-like, substantially fluid, or substantially solid. In some embodiments, the phase of the bilayer can be changed (e.g., by melting). Such a phase change can be caused by, for example, temperature changes, pH changes, compositional changes, storage, application, solvent changes, or any other factors.

[0133] In some embodiments, the lipid particles are lipid nanoparticles. As used herein, "lipid nanoparticle" refers to a nanoparticle comprising a plurality of lipid molecules. Furthermore, "nanoparticle" refers to a particle with a diameter of 1 nm to 1000 nm, but particularly 10 nm to 1000 nm. More particularly, the particle has a diameter of 30 nm to 500 nm, even more particularly 50 nm to 250 nm, especially particularly 50 nm to 200 nm, and most particularly 50 nm to 150 nm.

[0134] In some embodiments, the agent is contained within the aqueous phase of the lipid particle. In other embodiments, the agent is bound to the surface of the lipid particle (covalently or non-covalently). In some embodiments, the agent is wholly or partially embedded in the lipid bilayer of the lipid particle. In some embodiments, the agent is covalently bound to the lipid particle.

[0135] In some embodiments, the agent is a bioactive agent. Examples of bioactive agents include pharmaceutical compounds (e.g., small molecules), nucleic acids, oligonucleotides, dietary supplements, proteins, peptides, amino acids, etc. In some embodiments, the agent is a therapeutic agent.

[0136] In some embodiments, the agent is a nucleic acid. The nucleic acid agent can be double-stranded or single-stranded. Furthermore, the nucleic acid agent can be a linear or circular nucleic acid molecule. In some embodiments, the nucleic acid is DNA, RNA, or a non-natural nucleic acid. The RNA agent can be RNA encoding a peptide or non-coding RNA. The RNA agent can be mRNA, siRNA, miRNA, enhancer RNA, long non-coding RNA, etc. The nucleic acid can regulate gene expression, encode a peptide, or an aptamer, among other possibilities. In some embodiments, the nucleic acid is about 15 to about 30 nucleotides in length, for example, about 20-25 nucleotides (e.g., 20, 21, 22, 23, 24, or 25 nucleotides). In some embodiments, the nucleic acid is longer than about 30 nucleotides, for example, about 30, 35, 40, 45, 50, 60, 70, 80, 90, 100, 150, 200, 250, 300, 400, 500, 750, 1000, or more nucleotides.

[0137] In some implementations, the agent is a nutritional supplement, such as a vitamin, antioxidant, or dietary supplement.

[0138] In some embodiments, the agent is an imaging agent. Examples of imaging agents include, but are not limited to, chemical or biological dyes, fluorescent probes, radioactive isotopes, spin-labeled tracers, etc.

[0139] The lipid particles disclosed herein may contain one or more pharmaceutical agents (e.g., 2, 3, 4, 5, 6, 7, 8, 9, or 10 pharmaceutical agents).

[0140] As used herein, "zeta potential" refers to the potential at the interface between the bulk solution and the immobilized solvent and solute bound to the particles. The zeta potential reflects the degree of electrostatic repulsion between particles in the solution. Therefore, the zeta potential of lipid particles can be used as a predictor of particle stability in solution (e.g., anti-coagulation or aggregation). In some embodiments, the lipid particles of this disclosure have a zeta potential of less than -20 mV. In some embodiments, the lipid particles of this disclosure have a zeta potential of less than -10 mV. In some embodiments, the lipid particles of this disclosure have a zeta potential of less than -5 mV. In some embodiments, the lipid particles of this disclosure have a zeta potential of less than 0 mV. In some embodiments, the lipid particles of this disclosure have a zeta potential of less than 5 mV. In some embodiments, the lipid particles of this disclosure have a zeta potential of less than 10 mV. In some embodiments, the lipid particles of this disclosure have a zeta potential of less than 20 mV. In some embodiments, the lipid particles of this disclosure have a zeta potential of greater than -20 mV. In some embodiments, the lipid particles of this disclosure have a zeta potential of greater than -10 mV. In some embodiments, the lipid particles of this disclosure have a zeta potential greater than -5 mV. In some embodiments, the lipid particles of this disclosure have a zeta potential greater than 0 mV. In some embodiments, the lipid particles of this disclosure have a zeta potential greater than 5 mV. In some embodiments, the lipid particles of this disclosure have a zeta potential greater than 10 mV. In some embodiments, the lipid particles of this disclosure have a zeta potential greater than 20 mV. In some embodiments, the lipid particles have a zeta potential from -20 mV to 20 mV. In some embodiments, the lipid particles have a zeta potential from -10 mV to 20 mV. In some embodiments, the lipid particles have a zeta potential from 0 mV to 20 mV. In some embodiments, the lipid particles have a zeta potential from -20 mV to 10 mV. In some embodiments, the lipid particles have a zeta potential from -20 mV to 0 mV. In some embodiments, the lipid particles have a zeta potential from -10 mV to 10 mV. In some embodiments, the lipid particles have a zeta potential from 0 mV to 10 mV. In some embodiments, the lipid particles have a zeta potential of 5 mV to 10 mV. In some embodiments, the zeta potential is determined by light scattering, for example, using a Zetasizer Nano ZS (Malvern Instruments) in 0.1X PBS solution, as described in the examples.

[0141] As used herein, “pKa” refers to, for example, the apparent ionization constant of lipid nanoparticles. The pKa of the lipid nanoparticles of this disclosure can be from about 4 to about 9. In some embodiments, the pKa of the lipid nanoparticles of this disclosure is from about 5 to about 8. In some embodiments, the pKa of the lipid nanoparticles of this disclosure is from about 5.5 to about 7. Preferably, the pKa of the lipid nanoparticles of this disclosure is from about 6 to about 7. In some embodiments, the pKa is determined using a 2-(p-toluidine)naphthalene-6-sulfonic acid (TNS) assay, as described, for example, in the Journal of Controlled Release 107 (2005) 276–287 and examples by Heyes J. et al.

[0142] As used herein, encapsulation percentage refers to the amount of drug encapsulated within the particles of the composition, expressed as a percentage of the total amount of drug. In some embodiments, the encapsulation percentage is greater than 50%. In some embodiments, the encapsulation percentage is greater than 60%. In some embodiments, the encapsulation percentage is greater than 70%. In some embodiments, the encapsulation percentage is greater than 80%. In some embodiments, the encapsulation percentage is greater than 90%. In some embodiments, it is greater than 91%. In some embodiments, the encapsulation percentage is greater than 93%. In some embodiments, the encapsulation percentage is greater than 95%. In some embodiments, the encapsulation percentage is greater than 97%. In some embodiments, the encapsulation percentage is greater than 98%. In some embodiments, the encapsulation percentage is greater than 99%. In some embodiments, the encapsulation percentage is determined by a modified Quant-iT RiboGreen RNA assay (Thermo Fisher), for example, as described in Walsh C et al. Methods Mol Biol. 2014;1141:109-20 and the examples.

[0143] As used herein, the terms “polydispersity index,” “PDI,” and “dispersity” are used interchangeably and are measures of the non-uniformity of lipid particle size within a population of lipid particles. Lower values ​​indicate a more uniform size distribution. In some embodiments, the PDI of the lipid particles in the compositions described herein is less than 0.5. In some embodiments, the PDI of the lipid particles in the compositions described herein is less than 0.3. In some embodiments, the PDI of the lipid particles in the compositions described herein is less than 0.2. In some embodiments, the PDI of the lipid particles in the compositions described herein is less than 0.15. In some embodiments, the PDI of the lipid particles in the compositions described herein is less than 0.1. In some embodiments, the PDI of the lipid particles in the compositions described herein is less than 0.08. In some embodiments, the PDI of the lipid particles in the compositions described herein is less than 0.07. In some embodiments, the PDI of the lipid particles in the compositions described herein is less than 0.06. In some embodiments, the PDI of the lipid particles in the compositions described herein is less than 0.05. In some implementations, PDI is determined by dynamic light scattering, for example using Zetasizer Nano ZS (Malvern Instruments), as described in the examples.

[0144] As used herein, “Z-average” refers to the intensity-weighted average diameter of the lipid nanoparticles in the compositions described herein. In some embodiments, the Z-average particle size of the lipid nanoparticles of this disclosure can be from 10 nm to 500 nm. In some embodiments, the Z-average particle size of the lipid nanoparticles of this disclosure can be from 10 nm to 300 nm. In some embodiments, the Z-average particle size of the lipid nanoparticles of this disclosure can be from 10 nm to 200 nm. In some embodiments, the Z-average particle size of the lipid nanoparticles of this disclosure can be from 10 nm to 150 nm. In some embodiments, the Z-average particle size of the lipid nanoparticles of this disclosure can be from 10 nm to 100 nm. In some embodiments, the Z-average particle size of the lipid nanoparticles of this disclosure can be less than 300 nm. In some embodiments, the Z-average particle size of the lipid nanoparticles of this disclosure can be less than 250 nm. In some embodiments, the Z-average particle size of the lipid nanoparticles of this disclosure can be less than 200 nm. In some embodiments, the Z-average particle size of the lipid nanoparticles of this disclosure can be less than 150 nm. In some embodiments, the Z-mean particle size of the lipid nanoparticles of this disclosure may be less than 100 nm. In some embodiments, the Z-mean particle size of the lipid nanoparticles of this disclosure is determined by dynamic light scattering, for example using a ZetasizerNano ZS (Malvern Instruments), as described in the examples.

[0145] D. Ionizable lipids In some embodiments of the compositions disclosed herein, the compositions comprise ionizable lipids. In some embodiments, the ionizable lipids contain one or more groups that are ionic at physiological pH but may be uncharged at specific pH values ​​outside the physiological pH range. Typically, the ionizable lipids of this disclosure are cationic ionizable lipids having a cationic charge at physiological pH values. The ionizable cationic group may contain one or more protonable amines capable of forming a cationic group at physiological pH. The ionizable lipid compound may further comprise one or more lipid components, for example, having C1-C24... 30 Two or more fatty acids with alkyl or alkenyl carbon groups. In some embodiments of the compositions of this application, ionizable cationic lipids refer to lipid and lipid molecules having nitrogen atoms that can acquire a charge (pKa). These amino-containing molecules typically have 2 to 6 hydrophobic chains, usually alkyl or alkenyl, such as C1-C1. 30 Alkyl or alkenyl groups, but may have at least one or more than six tails.

[0146] The ionizable lipid component of the lipid nanoparticles of the present invention may account for 1 mol% to 99 mol% of the lipid molecules in the nanoparticles. In other embodiments, the ionizable lipid component may account for 5 mol% to 90 mol% of the lipid molecules in the lipid nanoparticles. In some embodiments, the ionizable lipid component accounts for 10 mol% to 75 mol%, 20 mol% to 75 mol%, 20 mol% to 60 mol%, or 20 mol% to 55 mol% of the lipid molecules in the lipid nanoparticles.

[0147] The ionizable lipids of this application may contain one or more asymmetrically substituted carbon or nitrogen atoms and may be separated in optically active or racemic form. Therefore, unless a specific stereochemical or isomeric form is specifically indicated, all chiral, diastereomeric, racemic, epimeric, and geometrical isomers of the chemical formula are contemplated. The ionizable lipids may exist as racemates and mixtures of racemates, as single enantiomers, mixtures of diastereomers, and single diastereomers. In some embodiments, a single diastereomer is obtained. The chiral center of the cationic ionizable lipids of this application may have an S or R configuration.

[0148] The ionizable lipids of this application may also have the following advantages: they may be more effective, less toxic, longer-lasting, more potent, produce fewer side effects, be more easily absorbed, and / or have better pharmacokinetic profiles (e.g., higher oral bioavailability and / or lower clearance), and / or have other useful pharmacological, physical, or chemical properties, whether for the indications described herein or other indications.

[0149] In some embodiments, the ionizable cationic lipid comprises a positively charged ammonium group at physiological pH and contains at least two hydrophobic groups. In some embodiments, the ammonium group is positively charged at a pH of about 6 to about 8. In some embodiments, the ionizable cationic lipid comprises at least two C1-C2 groups. 30 Alkyl or alkenyl groups.

[0150] In some embodiments, the ionizable lipids suitable for use in this disclosure are biodegradable. As used herein, "biodegradable" means the property of lipids or other compounds to be broken down into smaller groups, which are then metabolized, catabolized, excreted, and / or otherwise removed from the body through natural chemical and biological processes within the individual. Biodegradation can occur chemically without enzymatic action, for example, the breakdown of esters into carboxylates and alcohols in high or low pH environments. In other embodiments, biodegradation is catalyzed by enzymes, such as the breakdown of esters by esterases. Certain groups are more readily or rapidly biodegraded than others and contribute to the rate of biodegradation of larger molecules, such as lipids or lipid particles. The incorporation of biodegradable groups into the ionizable lipids results in faster metabolism and removal from the body after the active agent is delivered to the target region. Therefore, these ionizable lipids can have lower toxicity than similar ionizable lipids without biodegradable groups.

[0151] Examples of biodegradable groups applicable to this disclosure include, but are not limited to: -OC(O)-, -C(O)O-, -SC(O)-, -C(O)S-, -OC(S)-, -C(S)O-, -SS-, -C(Z) 3 )=N-、-N=C(Z 3 )-,-C(Z 3 )=NO-,-ON=C(Z 3 )-、-C(O)(NZ 3 )-,-N(Z 3 )C(O)-,-C(S)(NZ 3 )-,-N(Z 3 )C(O)-、-N(Z 3 )C(O)N(Z 3 -, -OC(O)O-, -OSi(Z) 3 )2O-、-C(O)(CZ 1 Z 2 )C(O)O- or -OC(O)(CZ 1 Z 2 )C(O)-; where Z 1 and Z 2 Independently, it is H, OH, alkyl, alkoxy, -NH2, alkylamino, or dialkylamino, and Z 3It is H or alkyl. Generally, compounds containing more biodegradable groups are considered more biodegradable themselves. In some embodiments, the biodegradable lipids of this disclosure contain two or more biodegradable groups. In some embodiments, the biodegradable lipids of this disclosure contain three or more biodegradable groups. In some embodiments, the biodegradable lipids of this disclosure contain four or more biodegradable groups. In some embodiments, the lipid is considered biodegradable even though it does not have any of the biodegradable groups listed above.

[0152] In some embodiments, the ionizable lipid used in this disclosure is a lipid of formula (I) or a pharmaceutically acceptable salt thereof: in X is -NR 1 -or -O-; R 1 It is a hydrogen atom, a hydrocarbon group with 6 to 24 atoms, or R 21 -L 1 -R 22 -; R 21 It is a hydrocarbon group having 1 to 24 carbon atoms; L 1 Is -O(CO)O-, -O(CO)-, -(CO)O-, -O- or ; R 22 It is a divalent hydrocarbon linking group having 1 to 18 carbon atoms; R 2 and R 3 Each is independently a hydrogen atom, a hydrocarbon group having 3 to 24 carbon atoms, or an R... 31 -L 1 -R 32 -; R 31 It is a hydrocarbon group having 1 to 24 carbon atoms; L 2 Is -O(CO)O-, -O(CO)-, -(CO)O-, -O- or ; R 32 It is a divalent hydrocarbon linking group having 1 to 18 carbon atoms; R 4 R 5 R 6 R 7 R 8 R 9 R 10 R 11 and R12 Each is independently a hydrogen atom or optionally R 33 Substituted alkyl groups having 1 to 18 carbon atoms, wherein the alkyl group is selected from R 4 and R 5 R 10 and R 5 R 5 and R 12 R 4 and R 6 R 5 and R 6 R 6 and R 7 R 6 and R 10 R 12 and R 7 and R 7 and R 8 One or more pairs of groups can be linked together to form a 4- to 7-membered ring of carbon and nitrogen atoms, said ring may contain oxygen atoms; R 33 It is a hydroxyl group, a carboxyl group, or -NR. 45 R 46 -O(CO)OR 41 -O(CO)-R 42 -(CO)OR 43 -OR 44 Or optionally R 34 Substituted aryl or heteroaryl groups; R 34 It is an alkyl group, hydroxy group, carboxyl group, or -NR group having 1 to 18 carbon atoms. 45 R 46 -O(CO)OR 41 -O(CO)-R 42 -(CO)OR 43 or -OR 44 ; R 41 R 42 R 43 R 44 R 45 and R 46 Each is independently a hydrocarbon group having 1 to 18 carbon atoms; and a, b, c, and d are each independent integers from 0 to 3, where a+b is 1 or greater, and c+d is 1 or greater.

[0153] For those from R 1 Hydrocarbon groups with 6 to 24 carbon atoms and those derived from R 2 and R 3The alkyl group has 3 to 24 carbon atoms, preferably an alkyl group, an alkenyl group, or an alkynyl group, and more preferably an alkyl group or an alkenyl group. The alkyl group can be straight-chain or branched, or it can be chain-like or cyclic. The alkyl group having 6 to 24 carbon atoms is preferably an alkyl group having 6 to 20 carbon atoms, and the alkyl group having 3 to 24 carbon atoms is more preferably an alkyl group having 6 to 20 carbon atoms. Specific examples include hexyl groups, heptyl groups, octyl groups, nonyl groups, decyl groups, undecyl groups, dodecyl groups, tridecyl groups, trimethyl dodecyl groups (preferably 3,7,11-trimethyl dodecyl groups), tetradecyl groups, pentadecyl groups, hexadecyl groups, tetramethyl hexadecyl groups (preferably 3,7,11,15-tetramethyl hexadecyl groups), heptadecanyl groups, octadecyl groups, nonadecanyl groups, eicosyl groups, etc. Alkenyl groups having 6 to 24 carbon atoms and alkenyl groups having 3 to 24 carbon atoms can be straight-chain or branched, or can be chain-like or cyclic. Alkenyl groups having 6 to 24 carbon atoms are preferably alkenyl groups having 6 to 20 carbon atoms, and alkenyl groups having 3 to 24 carbon atoms are more preferably alkenyl groups having 6 to 20 carbon atoms. Specific examples include hexenyl groups, heptenyl groups, octenyl groups, nonenyl groups, decenyl groups, undecenyl groups, dodecenyl groups, dodecadienyl groups, tridecenyl groups, tetradecenyl groups, pentadecenyl groups, hexadecenyl groups (preferably (Z)-hexadec-9-enyl groups), hexadecadienyl groups, heptadecanyl groups (preferably (Z)-heptadec-8-enyl groups), heptadecanadienyl groups (preferably (Z)-8-enyl groups), and heptadecanadienyl groups (preferably (Z)-8-enyl groups). Alkynyl groups having 6 to 24 carbon atoms are preferred to be 8,11-heptadecyl-8,11-dienyl groups, octadecyl-1 (preferably 9,12-dienyl groups), octadecyl-1 (preferably 9,12-dienyl groups), nonadecyl-1 (preferably 11-11-dienyl groups), and eicosyl-1 (preferably 11,14-dienyl groups). Alkynyl groups having 6 to 24 carbon atoms are preferred to be 6 to 20 carbon atoms, and alkynyl groups having 3 to 24 carbon atoms are more preferably 6 to 20 carbon atoms. Specifically, examples include hexynyl groups, heptyynyl groups, octyynyl groups, nonynyl groups, decyynyl groups, undecynyl groups, dodecynyl groups, tetradecynyl groups, pentadecynyl groups, hexadecimalyl groups, heptadecynyl groups, heptadecynyl groups, octadecynyl groups, etc. All of the above-mentioned alkenyl groups preferably have one or two double bonds. All of the above-mentioned ynyl groups preferably have one or two triple bonds.

[0154] For those from R21 and R 31The alkyl group has 1 to 24 carbon atoms, preferably an alkyl group, an alkenyl group, or an alkynyl group with 10 to 24 carbon atoms. The alkyl group with 10 to 24 carbon atoms can be straight-chain or branched, or it can be chain-like or cyclic. The alkyl group with 10 to 24 carbon atoms is preferably an alkyl group with 12 to 24 carbon atoms. Specific examples include decyl groups, undecyl groups, dodecyl groups, tridecyl groups, trimethyldodecyl groups (preferably 3,7,11-trimethyldodecyl groups), tetradecyl groups, pentadecyl groups, hexadecyl groups, tetramethylhexadecyl groups (preferably 3,7,11,15-tetramethylhexadecyl groups), heptadecanyl groups, octadecyl groups, 2-butylhexyl groups, 2-butyloctyl groups, 1-pentylhexyl groups, 2-pentylheptyl groups, 3-pentyloctyl groups, 1-hexylheptyl groups, and 1-hexylnonyl groups. Alkenyl groups, including 2-hexyloctyl, 2-hexyldecyl, 3-hexylnonyl, 1-heptyloctyl, 2-heptylnonyl, 2-heptylundecyl, 3-heptyldecyl, 1-octylnonyl, 2-octyldecyl, 2-octyldodecyl, 3-octylundecyl, 2-nonylundecyl, 3-nonyldodecyl, 2-decyldodecyl, 2-decyltetradecyl, 3-decyltridecyl, and 2-(4,4-dimethylpentan-2-yl)-5,7,7-trimethyloctyl, etc., can be straight-chain or branched, and can be chain-like or cyclic. Specific examples include decenyl groups, undecenyl groups, dodecenyl groups, dodecadienyl groups, tridecenyl groups (preferably (Z)-twelfthoc-8-enyl groups), tetradecenyl groups (preferably tetradecen-9-enyl groups), pentadecenyl groups (preferably (Z)-pentadec-8-enyl groups), hexadecenyl groups (preferably (Z)-hexadecen-9-enyl groups), hexadecadienyl groups, heptadecenyl groups (preferably (Z)-heptadecen-8-enyl groups), heptadecadienyl groups (preferably (8Z,11Z)-heptadecen-8,11-dienyl groups), octadecenyl groups (preferably (Z)-octadecen-9-enyl groups), and octadecadienyl groups (preferably (9Z,12Z)-octadecen-9,12-dienyl groups), etc. Alkynyl groups having 10 to 24 carbon atoms can be straight-chain or branched, or they can be chain-like or cyclic. Specific examples include decynyl groups, undecynyl groups, dodecynyl groups, tetradecynyl groups, pentadecynyl groups, hexadecimalyl groups, heptadecynyl groups, heptadecynyl groups, and octadecynyl groups. All of the above alkenyl groups preferably have one or two double bonds. All of the above ynyl groups preferably have one or two triple bonds.

[0155] For those from R 22 and R 32 The linkage group is a divalent hydrocarbon having 1 to 18 carbon atoms, preferably an alkylene group having 1 to 18 carbon atoms or an alkenyl group having 2 to 18 carbon atoms. The alkylene group having 1 to 18 carbon atoms can be straight-chain or branched, or chain-like or cyclic. The number of carbon atoms in the alkylene group is preferably 1 to 12, more preferably 1 to 10, and even more preferably 2 to 10. Specific examples include methylene, ethylene, trimethylene, tetramethylene, pentamethylene, hexamethylene, heptamethylene, octamethylene, nonamethylene, decamethylene, undecamethylene, dodecamethylene, etc. The alkenyl group having 2 to 18 carbon atoms can be straight-chain or branched, or chain-like or cyclic. The number of carbon atoms in the alkenyl group is preferably 1 to 12, and more preferably 2 to 10.

[0156] L 1 and L 2 The preferred embodiments are -O(CO)O-, -O(CO)-, and -(CO)O-. L 1 and L 2 More preferred embodiments are -O(CO)- and -(CO)O-.

[0157] From R 4 R 6 R 9 R 10 R 11 and R 12 The optionally substituted alkyl group having 1 to 18 carbon atoms can be straight-chain or branched, or chain-like or cyclic. The number of carbon atoms in the alkyl group is preferably 1 to 12. Specific examples include methyl groups, ethyl groups, propyl groups, isopropyl groups, cyclopropyl groups, butyl groups, isobutyl groups, tert-butyl groups, cyclobutyl groups, pentyl groups, cyclopentyl groups, hexyl groups, cyclohexyl groups, heptyl groups, octyl groups, nonyl groups, decyl groups, undecyl groups, dodecyl groups, etc. When the alkyl group has substituents, hydroxyl groups, carboxyl groups, or groups derived from -O(CO)OR are preferred. 41 -O(CO)-R 42 -(CO)OR 43 or -OR 44 The group represented, and more preferably -O(CO)-R 42 or-(CO)OR 43 The group indicated.

[0158] From R 5 R 7 and R 8The optionally substituted alkyl group having 1 to 18 carbon atoms can be straight-chain or branched, or chain-like or cyclic. The number of carbon atoms in the alkyl group is preferably 1 to 12, and more preferably 1 to 8. Specific examples include methyl groups, ethyl groups, propyl groups, isopropyl groups, cyclopropyl groups, butyl groups, isobutyl groups, tert-butyl groups, cyclobutyl groups, pentyl groups, cyclopentyl groups, hexyl groups, cyclohexyl groups, heptyl groups, octyl groups, nonyl groups, decyl groups, undecyl groups, dodecyl groups, etc. When the alkyl group has substituents, hydroxyl groups, carboxyl groups, or groups derived from -O(CO)OR are preferred. 41 -O(CO)-R 42 -(CO)OR 43 or -OR 44 The group represented, and more preferably -O(CO)-R 42 -(CO)OR 43 or -OR 44 The group indicated.

[0159] Examples of 4- to 7-membered rings that may contain carbon and nitrogen atoms, including azacyclic butane rings, pyrrolidine rings, piperidine rings, morpholine rings, and azacyclic heptane rings. The 4- to 7-membered rings are preferably 6-membered rings, and more preferably piperidine or morpholine rings.

[0160] When from R 4 R 5 R 6 R 7 R 8 R 9 R 10 R 11 and R 12 When an optionally substituted alkyl group having 1 to 18 carbon atoms is substituted or substituted with an unsubstituted aryl group, the number of carbon atoms in the aryl group is preferably 6 to 22, more preferably 6 to 18, and even more preferably 6 to 10. Specific examples of the aryl group include phenyl groups, naphthyl groups, anthraceneyl groups, phenanthrene groups, etc. For the substituents on the aryl group, alkyl groups, hydroxyl groups, carboxyl groups, and groups having 1 to 18 carbon atoms are preferred. 45 R 46 The amino group represented is either -O(CO)OR 41 -O(CO)-R 42 -(CO)OR 43 or -OR 44 The indicated group, and more preferably a hydroxyl group or a carboxyl group. Specific examples of substituted aryl groups include hydroxyphenyl groups, carboxyphenyl groups, etc.

[0161] When from R 4 R 5 R 6 R 7 R 8 R 9 R 10 R 11 and R 12 When an optionally substituted alkyl group having 1 to 18 carbon atoms is replaced by a substituted or unsubstituted heteroaryl group, the number of carbon atoms in the heteroaryl group is preferably 1 to 12, and more preferably 1 to 6. Specific examples of the heteroaryl group include pyridyl groups, pyrazolyl groups, imidazole groups, benzimidazole groups, thiazolyl groups, oxazolyl groups, etc. For the substituents on the heteroaryl group, alkyl groups, hydroxyl groups, carboxyl groups, and groups having 1 to 18 carbon atoms are preferred. 45 R 46 The amino group represented is either -O(CO)OR 41 -O(CO)-R 42 -(CO)OR 43 or -OR 44 The indicated group, and more preferably a hydroxyl or carboxyl group. Specific examples of substituted or unsubstituted heteroaryl groups include hydroxypyridinyl groups, carboxypyridinyl groups, pyridinone groups, etc.

[0162] For R 41 R 42 R 43 R 44 R 45 and R 46The hydrocarbon group represented is a hydrocarbon group having 1 to 18 carbon atoms, preferably an alkyl group having 1 to 18 carbon atoms, an alkenyl group having 2 to 18 carbon atoms, or an alkynyl group having 2 to 18 carbon atoms, and more preferably an alkyl group having 1 to 18 carbon atoms or an alkenyl group having 2 to 18 carbon atoms. The alkyl group having 1 to 18 carbon atoms may be straight-chain or branched, or may be chain-like or cyclic. The number of carbon atoms in the alkyl group is preferably 3 to 18, and more preferably 5 to 18. Specific examples include propyl groups, isopropyl groups, cyclopropyl groups, butyl groups, isobutyl groups, tert-butyl groups, cyclobutyl groups, pentyl groups, cyclopentyl groups, hexyl groups, cyclohexyl groups, heptyl groups, octyl groups, nonyl groups, decyl groups, undecyl groups, dodecyl groups, tridecyl groups, trimethyldodecyl groups (preferably 3,7,11-trimethyldodecyl groups), tetradecyl groups, pentadecyl groups, hexadecyl groups, heptadecanyl groups, octadecyl groups, etc. The alkenyl groups having 2 to 18 carbon atoms can be straight-chain or branched, or chain-like or cyclic. The number of carbon atoms in the alkenyl groups is preferably 3 to 18, and more preferably 5 to 18. Specific examples include allyl groups, isopentenyl groups, pentenyl groups, hexenyl groups, heptenyl groups, octenyl groups, nonenyl groups (preferably (Z)-2-nonenyl groups or (E)-2-nonenyl groups), decenyl groups, undecenyl groups, dodecenyl groups, dodecadienyl groups, tridecenyl groups (preferably (Z)-twelftho-8-enyl groups), tetradecenyl groups (preferably tetradectho-9-enyl groups), pentadecenyl groups (preferably... (Z)-pentadecano-8-enyl group, hexadecenoic group (preferably (Z)-hexadeco-9-enyl group), hexadecadienyl group, heptadecenoic group (preferably (Z)-heptadeco-8-enyl group), heptadecadienyl group (preferably (8Z,11Z)-heptadeco-8,11-dienyl group), octadecenoic group (preferably (Z)-octadeco-9-enyl group), octadecadienyl group (preferably (9Z,12Z)-octadeco-9,12-dienyl group), etc. The alkynyl group having 2 to 18 carbon atoms can be straight-chain or branched, or chain-like or cyclic. The number of carbon atoms in the alkynyl group is preferably 3 to 18, and more preferably 5 to 18. Specific examples include propynyl groups, butynyl groups, penynyl groups, hexynyl groups, hepynyl groups, octyynyl groups, nonynyl groups, decynyl groups, undecynyl groups, dodecynyl groups, tetradecynyl groups, pentadecynyl groups, hexadecimalyl groups, heptadecynyl groups, heptadecynyl groups, and octadecynyl groups.

[0163] When X represents -NR 1- At that time, R 1 Preferably, it represents a hydrocarbon group having 6 to 24 carbon atoms, or is composed of R 21 -L 1 -R 22 - indicates a group. When X represents R 21 -L 1 -R 22 - When, R is preferred 2 and R 3 One of them represents a hydrogen atom, and the other represents a hydrocarbon group having 6 to 24 carbon atoms, or a group composed of R. 31 -L 2 -R 32 - indicates a functional group.

[0164] When X represents -O-, R is preferred. 2 and R 3 Each independently represents a hydrocarbon group having 6 to 24 carbon atoms, or a hydrocarbon group composed of R. 31 -L 2 -R 32 - indicates a functional group.

[0165] Preferably, R 4 R 6 R 9 R 10 R 11 and R 12 Each represents a hydrogen atom.

[0166] R 5 Preferably, it contains hydrogen atoms, an alkyl group having 1 to 18 carbon atoms, or a -O(CO)-R group. 42 or-(CO)OR 43 Alkyl groups having 1 to 18 carbon atoms that are substituted, alkyl groups having 1 to 18 carbon atoms that are substituted with aryl groups, or alkyl groups having 1 to 18 carbon atoms that are substituted with hydroxyl groups. Specifically, R 5 Preferably, it has an alkyl group having 1 to 18 carbon atoms, which can be converted to -O(CO)-R 42 or-(CO)OR 43 The substituted alkyl group having 1 to 18 carbon atoms, the alkyl group having 1 to 12 carbon atoms that can be substituted with an aryl group, or the alkyl group having 1 to 8 carbon atoms that can be substituted with a hydroxyl group, and more preferably the alkyl group having 1 to 18 carbon atoms, or the alkyl group that can be substituted with -O(CO)-R 42 or-(CO)OR 43 Substituted alkyl groups having 1 to 18 carbon atoms. In R 5 In the case of an alkyl group, R 5 It can be connected to R 4 R6 R 10 or R 12 To form rings that can contain oxygen atoms.

[0167] R 7 and R 8 Preferably, each of the following groups independently represents a hydrogen atom, a hydrocarbon group having 1 to 18 carbon atoms, and is optionally prefixed with -O(CO)-R. 42 or-(CO)OR 43 The substituted alkyl group having 1 to 18 carbon atoms, the alkyl group having 1 to 8 carbon atoms that can be substituted with an aryl group, or the alkyl group having 1 to 8 carbon atoms that can be substituted with a hydroxyl group. Alternatively, preferably, R 7 and R 8 They connect to each other to form 4 to 7-membered rings that can contain oxygen atoms, carbon atoms, and nitrogen atoms.

[0168] R 5 Not with R 7 or R 8 Connect, and not with R 7 or R 8 A ring is formed.

[0169] The sum of a+b is preferably 1 or 2, and more preferably 1. The sum of c+d is preferably 1 or 2, and more preferably 1.

[0170] The lipids represented by formula (I) and their preparation methods are described in US2021 / 0085604 and US2022 / 0273817, the contents of which are incorporated herein by reference in their entirety.

[0171] The compound represented by formula (I) is preferably the compound represented by formula (III) or a pharmaceutically acceptable salt thereof: in R 2 and R 3 Each is independently a hydrogen atom, a hydrocarbon group having 3 to 24 carbon atoms, or R 31 -L 2 -R 32 -; R 31 It is a hydrocarbon group having 1 to 24 carbon atoms; L 2 Is -O(CO)O-, -O(CO)-, -(CO)O-, -O- or ; R 32 It is a divalent hydrocarbon linking group having 1 to 18 carbon atoms; R 5 It is a hydrogen atom or optionally R33 Substituted alkyl groups having 1 to 18 carbon atoms; R 7 and R 8 Each is independently a hydrogen atom or optionally R 33 Substituted alkyl groups having 1 to 18 carbon atoms; R 33 It is a hydroxyl group, a carboxyl group, or -NR. 45 R 46 -O(CO)OR 41 -O(CO)-R 42 -(CO)OR 43 -OR 44 Or optionally R 34 Substituted aryl or heteroaryl groups; R 34 It is an alkyl group, hydroxy group, carboxyl group, or -NR group having 1 to 18 carbon atoms. 45 R 46 -O(CO)OR 41 -O(CO)-R 42 -(CO)OR 43 or -OR 44 ; R 41 R 42 R 43 R 44 R 45 and R 46 Each is independently a hydrocarbon group having 1 to 18 carbon atoms; and e is an integer selected from 2 or 3.

[0172] Examples of salts containing basic groups include salts with inorganic acids such as hydrochloric acid, hydrobromic acid, nitric acid, and sulfuric acid; salts with organic carboxylic acids such as formic acid, acetic acid, citric acid, oxalic acid, fumaric acid, maleic acid, succinic acid, malic acid, tartaric acid, aspartic acid, trichloroacetic acid, and trifluoroacetic acid; and salts with sulfonic acids such as methanesulfonic acid, benzenesulfonic acid, p-toluenesulfonic acid, mesitylenesulfonic acid, and naphthalenesulfonic acid.

[0173] Examples of salts with acidic groups include salts with alkali metals such as sodium and potassium; salts with alkaline earth metals such as calcium and magnesium; ammonium salts; and salts with nitrogen-containing organic bases such as trimethylamine, triethylamine, tributylamine, pyridine, N,N-dimethylaniline, N-methylpiperidine, N-methylmorpholine, diethylamine, dicyclohexylamine, procaine, dibenzylamine, N-benzyl-β-phenylethylamine, L-ephenamine, and N,N'-dibenzylethylenediamine.

[0174] Of the salts mentioned above, those that are pharmacologically acceptable are preferred.

[0175] Lipids represented by formula (III) and methods for their preparation are described in US2021 / 0085604 and US2022 / 0273817. The entire contents of each disclosure are incorporated herein by reference.

[0176] Exemplary lipids represented by formula (III) include: FL-D: ; FL-B: ; FL-A: ; And FL-C: .

[0177] In some embodiments, the ionizable lipids suitable for use in this disclosure are ionizable lipids of formula (II) or pharmaceutically acceptable salts thereof: in R 51 and R 52 Each independently is arbitrarily R 35 Substituted hydrocarbon groups having 1 to 21 carbon atoms; R 35 It is a hydroxyl group, -G 20 -CH(R 55 (R) 56 ), -N(R 58 (R) 59 ) or -G 20 -R 60 ; G 20 It is -(CO)O- or -O(CO)-; R 55 and R 56 Each is independently hydrogen or a hydrocarbon group having 1 to 18 carbon atoms; R 58 and R 59 Each independently is hydrogen or optionally R 36 Substituted cyclic hydrocarbon groups having 3 to 6 carbon atoms; R 60 It is a hydrocarbon group having 1 to 18 carbon atoms; R 36 It is -N(R) 61 (R) 62 ) or -G 20 -R 65 ; R61 and R 62 Each is independently hydrogen or a cyclic hydrocarbon group having 3 to 6 carbon atoms; R 65 It is a hydrocarbon group with 1 to 18 carbon atoms, -L 40 -CH(R 66 (R) 67 ) or -G 20 -R 66 ; L 40 It is a divalent hydrocarbon group containing 1 to 6 carbon atoms; R 66 and R 67 Each is independently a hydrocarbon group containing 1 to 10 carbon atoms, or an alkoxy group containing 1 to 10 carbon atoms; L 10 It is a divalent hydrocarbon group containing 1 to 18 carbon atoms; G 30 -S(CO)N(R) 64 )-; R 64 Yes -L 30 -G 20 -CH(R 55 (R) 56 ); a' is 0 or 1; G 10 It is G 20 -O(CO)O- or -N(R) 63 )C(O)-; R 63 It is a hydrocarbon group containing 1 to 18 carbon atoms; L 20 It is a divalent hydrocarbon group containing 1 to 6 carbon atoms; b' is 0 or 1; R 53 R 54 and R 57 Each independently is hydrogen or optionally R 36 Substituted hydrocarbon groups containing 1 to 18 carbon atoms; and L 30 It is a single bond or a hydrocarbon group containing 1 to 18 carbon atoms.

[0178] The compound represented by formula (II) can be represented by formula (IIa). in R 51 and R 52 Each independently is arbitrarily R 35Substituted hydrocarbon groups having 1 to 21 carbon atoms; R 35 It is a hydroxyl group or -G 20 -CH(R 55 (R) 56 ); G 20 It is -(CO)O- or -O(CO)-; R 55 and R 56 Each is independently hydrogen or a hydrocarbon group having 1 to 18 carbon atoms; L 10 It is a divalent hydrocarbon group containing 1 to 18 carbon atoms; G 10 It is -(CO)O- or -O(CO)-; and R 53 R 54 and R 57 Each can be either hydrogen or a hydrocarbon group containing 1 to 18 carbon atoms.

[0179] The compound represented by formula (II) can be represented by formula (IIb). in R 51 and R 52 Each is an independent hydrocarbon group having 1 to 21 carbon atoms; L 10 It is a divalent hydrocarbon group containing 1 to 18 carbon atoms; G 10 is -O(CO)O-; L 20 It is a divalent hydrocarbon group containing 1 to 6 carbon atoms; R 53 R 54 and R 57 Each independently is hydrogen or optionally R 36 Substituted hydrocarbon groups containing 1 to 18 carbon atoms; R 36 Yes-O(CO)-R 65 ; R 65 It is a hydrocarbon group having 1 to 18 carbon atoms, or -L 40 -CH(R 66 (R) 67 ); L 40 It is a divalent hydrocarbon group containing 1 to 6 carbon atoms; and R 66 and R 67Each is an alkoxy group containing 1 to 10 carbon atoms.

[0180] The compound represented by formula (II) can be represented by formula (IIc). in R 51 and R 52 Each is an independent hydrocarbon group having 1 to 21 carbon atoms; L 10 It is a divalent hydrocarbon group containing 1 to 18 carbon atoms; G 10 It is -N(R) 63 )C(O)-; R 63 It is a hydrocarbon group containing 1 to 18 carbon atoms; R 53 R 54 and R 57 Each independently is hydrogen or optionally R 36 Substituted hydrocarbon groups containing 1 to 18 carbon atoms; R 36 Yes-(CO)OR 65 ; R 65 Yes -L 40 -CH(R 66 (R) 67 ); L 40 It is a divalent hydrocarbon group containing 1 to 6 carbon atoms; and R 66 and R 67 Each is an independent hydrocarbon group containing 1 to 10 carbon atoms.

[0181] The compound represented by formula (II) can be represented by formula (IId). in R 51 and R 52 Each is an independent hydrocarbon group having 1 to 21 carbon atoms; L 10 It is a divalent hydrocarbon group containing 1 to 18 carbon atoms; G 30 -S(CO)NR 64 -; R 64 Yes -L 30 -G 20 -CH(R 55 (R) 56 ); L 30 It is a single bond or a hydrocarbon group containing 1 to 18 carbon atoms; G 20 is-(CO)O-; R 55 and R 56 Each is independently hydrogen or a hydrocarbon group having 1 to 18 carbon atoms; G 10 It is -(CO)O-; and R 53 R 54 and R 57 Each can be either hydrogen or a hydrocarbon group containing 1 to 18 carbon atoms.

[0182] In some embodiments, the compounds of formula (II), (IIa), (IIb), (IIc) or (IId) may be in the form of salts.

[0183] In some embodiments, the ionizable lipids suitable for use in this disclosure are ionizable lipids of formula (VII) or pharmaceutically acceptable salts thereof: in R 1 and R 2 Each is independently a hydrocarbon group having 1 to 18 carbon atoms, and R 3 It is a hydrocarbon group having 2 to 8 carbon atoms, wherein R 1 R 2 and R 3 The hydrocarbon groups represented are each independently and optionally surrounded by one or more groups selected from -OH, COOH, -NR. 51 R 52 -OC(O)OR 53 -C(O)OR 54 -OC(O)-R 55 and -OR 56 Substituents of the substituents; R 4 It is a hydrocarbon group having 1 to 8 carbon atoms; R 5 and R 6 Each is independently a hydrocarbon group having 1 to 8 carbon atoms, or -R 8 -L 1 -R 9 Exclude R 5 and R 6 Both cases involve hydrocarbon groups having 1 to 8 carbon atoms; R 7 Yes -R 10 -L 2 -R 11-L 3 -R 12 ; R 51 and R 52 Each is an independent hydrocarbon group having 1 to 8 carbon atoms; R 53 R 54 R 55 and R 56 Each is an independent hydrocarbon group having 1 to 24 carbon atoms; By R 53 R 54 R 55 and R 56 Each of the hydrocarbon groups represented is independently and optionally surrounded by an aryl group having 6 to 20 carbon atoms or -SR 58 The aryl group having 6 to 20 carbon atoms is optionally replaced by -OH, COOH, or -NR. 51 R 52 -OC(O)OR 53 -C(O)OR 54 -OC(O)-R 55 -OR 56 or -R 59 -R 57 Replace; among them R 58 and R 59 Each is an independent hydrocarbon group having 1 to 12 carbon atoms; R 57 -OH, COOH, -NR 61 R 62 -OC(O)OR 63 -C(O)OR 64 -OC(O)-R 65 or -OR 66 ; R 61 and R 62 Each is an independent hydrocarbon group having 1 to 8 carbon atoms; R 63 R 64 R 65 and R 66 Each is an independent hydrocarbon group having 1 to 24 carbon atoms; By R 63 R 64 R 65 and R 66 Each of the hydrocarbon groups represented is independently and optionally surrounded by an aryl group having 6 to 20 carbon atoms or -SR 68The aryl group having 6 to 20 carbon atoms can be replaced by -OH, COOH, -NR, etc. 61 R 62 -OC(O)OR 63 -C(O)OR 64 -OC(O)-R 65 -OR 66 or -R 69 -R 67 replace; R 68 and R 69 Each is an independent hydrocarbon group having 1 to 12 carbon atoms; R 67 -OH, COOH, -NR 61 R 62 -OC(O)OR 63 -C(O)OR 64 -OC(O)-R 65 or -OR 66 ; L 1 L 2 and L 3 Each can be independently represented as -OC(O)O-, -C(O)O-, -OC(O)-, or -O-; R 8 It is a hydrocarbon group having 1 to 12 carbon atoms; R 9 It is a hydrocarbon group having 1 to 24 carbon atoms; R 10 It is a hydrocarbon group having 1 to 8 carbon atoms; R 11 It is a hydrocarbon group having 1 to 24 carbon atoms; R 12 It is a hydrocarbon group having 1 to 24 carbon atoms; By R 9 and R 12 The hydrocarbon groups represented are each independently and optionally replaced by aryl groups, -OC(O)OR 53 -C(O)OR 54 -OC(O)-R 55 or -SR 58 Replace; and By R 11 The hydrocarbon groups represented are each independently and optionally labeled -OC(O)OR 53 -C(O)OR 54 or -OC(O)-R 55 replace.

[0184] The hydrocarbon group having 1 to 24 carbon atoms, the hydrocarbon group having 1 to 18 carbon atoms, the hydrocarbon group having 1 to 12 carbon atoms, the hydrocarbon group having 2 to 8 carbon atoms, and the hydrocarbon group having 1 to 8 carbon atoms are each preferably alkyl groups, alkenyl groups, or alkynyl groups.

[0185] The alkyl group may be straight-chain or branched, or it may be chain-like or cyclic. Specifically, examples of the alkyl group include methyl, ethyl, propyl, isopropyl, cyclopropyl, butyl, isobutyl, tert-butyl, cyclobutyl, pentyl, cyclopentyl, hexyl, cyclohexyl, heptyl, octyl, nonyl, decyl, undecyl, dodecyl, tridecyl, trimethyldodecyl (preferably 3,7,11-trimethyldodecyl), tetradecyl, pentadecyl, hexadecyl, tetramethylhexadecyl (preferably 3,7,11,15-tetramethylhexadecyl), heptadecanyl, octadecyl, 2-butylhexyl, and 2-butyloctyl. The group includes 1-pentylhexyl group, 2-pentylheptyl group, 3-pentyloctyl group, 1-hexylheptyl group, 1-hexylnonyl group, 2-hexyloctyl group, 2-hexyldecyl group, 3-hexylnonyl group, 1-heptyloctyl group, 2-heptylnonyl group, 2-heptylundecyl group, 3-heptyldecyl group, 1-octylnonyl group, 2-octyldecyl group, 2-octyldodecyl group, 3-octylundecyl group, 2-nonylundecyl group, 3-nonyldodecyl group, 2-decyldodecyl group, 2-decyltetradecyl group, 3-decyltridecyl group, 2-(4,4-dimethylpent-2-yl)-5,7,7-trimethyloctyl group, etc.

[0186] The alkenyl group can be straight-chain or branched, or it can be chain-like or cyclic. Specifically, examples of the alkenyl group include allyl group, isopentenyl group, pentenyl group, hexenyl group, heptenyl group, octenyl group, nonenyl group (preferably (Z)-2-nonenyl group or (E)-2-nonenyl group), decenyl group, undecylenyl group, dodecylenyl group, dodecyl dienyl group, tridecylenyl group (preferably (Z)-twelfthocarbon-8-enyl group), tetradecylenyl group (preferably tetradecyl-9-enyl group), pentadecylenyl group (preferably (… (Z)-pentadecano-8-enyl group, hexadecyl group (preferably (Z)-hexadeco-9-enyl group), hexadecadienyl group, heptadecano-enyl group (preferably (Z)-heptadecano-8-enyl group), heptadecano-dienyl group (preferably (8Z,11Z)-heptadecano-8,11-dienyl group), octadecyl group (preferably (Z)-octadeco-9-enyl group), octadecyl dienyl group (preferably (9Z,12Z)-octadeco-9,12-dienyl group), etc. The aforementioned alkenyl groups preferably have one or two double bonds.

[0187] The alkynyl group can be straight-chain or branched, or it can be chain-like or cyclic. Specifically, examples of alkynyl groups include propynyl groups, butynyl groups, pentyynyl groups, hexynyl groups, heptyynyl groups, octyynyl groups, nonynyl groups, decynyl groups, undecylyl groups, dodecylyl groups, tetradecyl groups, pentadecyl groups, hexadecimalyl groups, heptadecyl groups, heptadecyl groups, and octadecyl groups. Preferably, the aforementioned alkynyl groups have one or two triple bonds.

[0188] R 69 Preferably, the alkylene group has 1 to 12 carbon atoms or the alkenyl group has 2 to 12 carbon atoms. The alkylene group with 1 to 12 carbon atoms and the alkenyl group with 2 to 12 carbon atoms can be straight-chain or branched, or can be chain-like or cyclic. Specifically, examples include methylene, ethylene, trimethylene, tetramethylene, pentamethylene, hexamethylene, heptamethylene, octamethylene, nonamethylene, decamethylene, undecamethylene, etc.

[0189] The aryl group preferably has 6 to 20 carbon atoms, more preferably 6 to 18 carbon atoms, and even more preferably 6 to 10 carbon atoms. Specifically, examples of the aryl group include phenyl groups, naphthyl groups, anthraceneyl groups, phenanthrene groups, etc.

[0190] R 1 and R 2Each is preferably a hydrocarbon group having 1 to 12 carbon atoms, more preferably a hydrocarbon group having 1 to 6 carbon atoms, and even more preferably a hydrocarbon group having 1 to 3 carbon atoms.

[0191] R 3 Preferably, it is a hydrocarbon group having 2 to 6 carbon atoms, and more preferably a hydrocarbon group having 2 to 4 carbon atoms.

[0192] Preferably, R 1 R 2 and R 3 Each of the hydrocarbon groups represented can be independently and optionally replaced by -OH.

[0193] L 1 and L 3 Each is preferably -C(O)O- or -OC(O)-.

[0194] L 2 Preferably, it is -OC(O)O-, -C(O)O-, or -OC(O)-.

[0195] R 8 Preferably, it is a hydrocarbon group having 1 to 10 carbon atoms, and more preferably a hydrocarbon group having 1 to 8 carbon atoms.

[0196] R 9 Preferably, it is a hydrocarbon group having 1 to 20 carbon atoms, and more preferably a hydrocarbon group having 1 to 16 carbon atoms.

[0197] R 11 Preferably, it is a hydrocarbon group having 1 to 16 carbon atoms, and more preferably a hydrocarbon group having 1 to 9 carbon atoms.

[0198] R 12 Preferably, it is a hydrocarbon group having 1 to 20 carbon atoms, and more preferably a hydrocarbon group having 1 to 16 carbon atoms.

[0199] Preferably, R 9 and R 12 The hydrocarbon groups represented are each independently and optionally surrounded by an aryl group or -SR. 58 Replace, where R 58 Preferably, it is a hydrocarbon group having 1 to 8 carbon atoms.

[0200] Preferably, R 11 The hydrocarbon group represented is optionally replaced by -C(O)OR 55 or -OC(O)-R 56 Replace, where R 55 and R 56 Each is an independent hydrocarbon group having 1 to 16 carbon atoms.

[0201] Preferably, R55 and R 56 Each of the hydrocarbon groups represented is independently and optionally surrounded by an aryl group having 6 to 20 carbon atoms or -SR 58 replace.

[0202] In some embodiments, the ionizable lipids suitable for use in this disclosure are ionizable lipids of formula (VIII) or pharmaceutically acceptable salts thereof: in R 1 and R 2 Each is independently a hydrocarbon group having 1 to 18 carbon atoms, and R 3 It is a hydrocarbon group having 2 to 8 carbon atoms, wherein R 1 R 2 and R 3 The hydrocarbon groups represented are each independently and optionally converted by -OH, COOH, or -NR. 51 R 52 -OC(O)OR 53 -C(O)OR 54 -OC(O)-R 55 or -OR 56 replace; R 4 It is a hydrocarbon group having 1 to 8 carbon atoms; R 5 and R 6 Each is independently a hydrocarbon group having 1 to 8 carbon atoms, or -R 8 -L 1 -R 9 Exclude R 5 and R 6 Both cases involve hydrocarbon groups having 1 to 8 carbon atoms; L 1 and L 4 Each can be independently represented as -OC(O)O-, -C(O)O-, -OC(O)-, or -O-; R 8 It is a hydrocarbon group having 1 to 12 carbon atoms; R 9 It is a hydrocarbon group having 1 to 24 carbon atoms, wherein R 9 The hydrocarbon group represented may optionally be replaced by an aryl group, -OC(O)OR 53 -C(O)OR 54 -OC(O)-R 55 or -SR 58 replace; R 51 and R 52Each is an independent hydrocarbon group having 1 to 8 carbon atoms; R 21 R 23 R 25 R 53 R 54 R 55 and R 56 Each is an independent hydrocarbon group having 1 to 24 carbon atoms; By R 53 R 54 R 55 and R 56 Each of the hydrocarbon groups represented is independently and optionally surrounded by an aryl group having 6 to 20 carbon atoms or -SR 58 The aryl group having 6 to 20 carbon atoms is optionally replaced by -OH, COOH, or -NR. 51 R 52 -OC(O)OR 53 -C(O)OR 54 -OC(O)-R 55 -OR 56 or -R 59 -R 57 replace; R 58 and R 59 Each is an independent hydrocarbon group having 1 to 12 carbon atoms; R 57 -OH, COOH, -NR 51 R 52 -OC(O)OR 53 -C(O)OR 54 -OC(O)-R 55 or -OR 56 ; R 13 It is a hydrocarbon group having 1 to 8 carbon atoms; R 14 Yes -R 15 -L 5 -R 16 , where R 15 It is a hydrocarbon group having 1 to 24 carbon atoms, L 5 It is -OC(O)O-, -C(O)O-, -OC(O)-, or -O-, and R 16 It is a hydrocarbon group having 1 to 24 carbon atoms; By R 15 The hydrocarbon group representing 1 to 24 carbon atoms is optionally represented by -OC(O)OR 53 -C(O)OR 54 or -OC(O)-R55 Replace; and By R 16 The hydrocarbon group having 1 to 24 carbon atoms is optionally replaced by an aryl group having 6 to 20 carbon atoms, -OC(O)OR 53 -C(O)OR 54 -OC(O)-R 55 or -SR 58 replace.

[0203] In equation (VIII), R 1 and R 2 Each is preferably a hydrocarbon group having 1 to 12 carbon atoms, more preferably a hydrocarbon group having 1 to 6 carbon atoms, and even more preferably a hydrocarbon group having 1 to 3 carbon atoms.

[0204] R 3 Preferably, it is a hydrocarbon group having 2 to 6 carbon atoms, and more preferably a hydrocarbon group having 2 to 4 carbon atoms.

[0205] Preferably, R 1 R 2 and R 3 Each of the hydrocarbon groups represented can be independently and optionally replaced by -OH.

[0206] L 1 Preferably, it is -C(O)O- or -OC(O)-.

[0207] R 8 Preferably, it is a hydrocarbon group having 1 to 10 carbon atoms, and more preferably a hydrocarbon group having 1 to 8 carbon atoms.

[0208] R 9 Preferably, it is a hydrocarbon group having 1 to 18 carbon atoms, and is composed of R 9 The indicated hydrocarbon group is optionally replaced by an aryl group having 6 to 20 carbon atoms or -SR 58 replace.

[0209] R 14 Preferred is -R 15 -L 5 -R 16 , where R 15 It is a hydrocarbon group having 1 to 18 carbon atoms, L 5 It is -OC(O)O-, and R 16 It is a hydrocarbon group having 1 to 18 carbon atoms.

[0210] Preferably, R 15 The hydrocarbon group represented by 1 to 18 carbon atoms is optionally replaced by -C(O)OR 55 or -OC(O)-R 56 Replace. R55 and R 56 Each is independently a hydrocarbon group having 1 to 16 carbon atoms, and is composed of R 55 and R 56 Each of the hydrocarbon groups represented is independently and optionally surrounded by an aryl group having 6 to 20 carbon atoms or -SR 58 replace.

[0211] Preferably, R 16 The hydrocarbon group having 1 to 18 carbon atoms is optionally replaced by an aryl group or -SR. 58 replace.

[0212] In some embodiments, the ionizable lipids suitable for use in this disclosure are ionizable lipids of formula (IX) or pharmaceutically acceptable salts thereof: in R 1 and R 2 Each is independently a hydrocarbon group having 1 to 18 carbon atoms, and R 3 It is a hydrocarbon group having 2 to 8 carbon atoms, wherein R 1 R 2 and R 3 The hydrocarbon groups represented are optionally replaced by -OH, COOH, or -NR. 51 R 52 -OC(O)OR 53 -C(O)OR 54 -OC(O)-R 55 or -OR 56 replace; R 4 and R 8 Each is independently a hydrocarbon having 1 to 8 carbon atoms; R 21 and R 22 Each is independently a hydrocarbon group having 1 to 18 carbon atoms; R 23 and R 24 Each is an independent hydrocarbon group having 1 to 12 carbon atoms; R 25 and R 26 Each is an independent hydrocarbon group having 1 to 24 carbon atoms; L 21 and L 22 Each can be independently represented as -OC(O)O-, -C(O)O-, -OC(O)-, or -O-; By R 25 and R 26The hydrocarbon groups represented are each independently and optionally replaced by an aryl group having 6 to 20 carbon atoms, -OC(O)OR 53 -C(O)OR 54 -OC(O)-R 55 or -SR 58 Substitution, wherein the aryl group having 6 to 20 carbon atoms is optionally replaced by OH, COOH, -NR 51 R 52 -OC(O)OR 53 -C(O)OR 54 -OC(O)-R 55 -OR 56 or -R 59 -R 57 Replace, and R 51 and R 52 Each is an independent hydrocarbon group having 1 to 8 carbon atoms; R 53 R 54 R 55 and R 56 Each is independently a hydrocarbon group having 1 to 18 carbon atoms; R 57 -OH, COOH, -NR 51 R 52 -OC(O)OR 53 -C(O)OR 54 -OC(O)-R 55 or -OR 56 ;and R 58 and R 59 Each is independently a hydrocarbon group having 1 to 12 carbon atoms. In formula (IX), R 1 and R 2 Each is preferably a hydrocarbon group having 1 to 12 carbon atoms, more preferably a hydrocarbon group having 1 to 6 carbon atoms, and even more preferably a hydrocarbon group having 1 to 3 carbon atoms. (R) 1 and R 2 The hydrocarbon groups represented are preferably each independently and optionally substituted with -OH, and more preferably are hydrocarbons without substituents.

[0213] R 3 Preferably, it is a hydrocarbon group having 2 to 6 carbon atoms, and more preferably a hydrocarbon group having 2 to 4 carbon atoms.

[0214] R 21 and R 22Each is preferably a hydrocarbon group having 1 to 12 carbon atoms, more preferably a hydrocarbon group having 1 to 8 carbon atoms, and even more preferably a hydrocarbon group having 1 to 6 carbon atoms.

[0215] R 23 and R 24 Each is preferably a hydrocarbon group having 1 to 10 carbon atoms, and more preferably a hydrocarbon group having 1 to 8 carbon atoms.

[0216] R 25 and R 26 Each is preferably a hydrocarbon group having 1 to 20 carbon atoms, more preferably a hydrocarbon group having 1 to 16 carbon atoms, and even more preferably a hydrocarbon group having 1 to 12 carbon atoms.

[0217] L 21 and L 22 Each is preferably -C(O)O- or -OC(O)-.

[0218] In some embodiments, the ionizable lipids suitable for use in this disclosure are ionizable lipids of formula (X) or pharmaceutically acceptable salts thereof: in R 1 and R 2 Each is independently a hydrocarbon group having 1 to 18 carbon atoms, and R 3 It is a hydrocarbon group having 2 to 8 carbon atoms, wherein R 1 R 2 and R 3 The hydrocarbon groups represented are each independently and optionally converted by -OH, COOH, or -NR. 51 R 52 -OC(O)OR 53 -C(O)OR 54 -OC(O)-R 55 or -OR 56 replace; R 4 and R 8 Each is an independent hydrocarbon group having 1 to 8 carbon atoms; R 31 R 32 R 33 and R 34 Each is independently a hydrocarbon group having 1 to 12 carbon atoms. R 35 R 36 R 37 and R 38 Each is an independent hydrocarbon group having 1 to 24 carbon atoms; L31 L 32 L 33 and L 34 Each can be independently represented as -OC(O)O-, -C(O)O-, -OC(O)-, or -O-; By R 35 R 36 R 37 and R 38 The hydrocarbon groups represented are each independently and optionally replaced by an aryl group having 6 to 20 carbon atoms, -OC(O)OR 53 -C(O)OR 54 -OC(O)-R 55 or SR 58 Substitution, wherein the aryl group having 6 to 20 carbon atoms is optionally replaced by OH, COOH, -NR 51 R 52 -OC(O)OR 53 -C(O)OR 54 -OC(O)-R 55 -OR 56 or -R 59 -R 57 Replace, and R 51 and R 52 Each is an independent hydrocarbon group having 1 to 8 carbon atoms; R 53 R 54 R 55 and R 56 Each is independently a hydrocarbon group having 1 to 18 carbon atoms; R 57 -OH, COOH, -NR 51 R 52 -OC(O)OR 53 -C(O)OR 54 -OC(O)-R 55 or -OR 56 And R 58 and R 59 Each is an independent hydrocarbon group having 1 to 12 carbon atoms.

[0219] In equation (X), R 1 and R 2 Each is preferably a hydrocarbon group having 1 to 12 carbon atoms, more preferably a hydrocarbon group having 1 to 6 carbon atoms, and even more preferably a hydrocarbon group having 1 to 3 carbon atoms. (R) 1 and R 2The hydrocarbon groups represented are preferably each independently and optionally substituted with -OH, and more preferably are hydrocarbons without substituents.

[0220] R 3 Preferably, it is a hydrocarbon group having 2 to 6 carbon atoms, and more preferably a hydrocarbon group having 2 to 4 carbon atoms.

[0221] R 31 R 32 R 33 and R 34 Each is preferably a hydrocarbon group having 1 to 10 carbon atoms, more preferably a hydrocarbon group having 1 to 8 carbon atoms, and even more preferably a hydrocarbon group having 1 to 3 carbon atoms.

[0222] R 35 R 36 R 37 and R 38 Each is preferably a hydrocarbon group having 1 to 20 carbon atoms, more preferably a hydrocarbon group having 1 to 16 carbon atoms, and even more preferably a hydrocarbon group having 1 to 12 carbon atoms. Preferably, it is composed of R... 35 R 36 R 37 and R 38 The hydrocarbon groups represented are each independently surrounded by an aryl group having 6 to 20 carbon atoms or an SR group. 58 And more preferably -SR 58 replace.

[0223] R 35 R 36 R 37 and R 38 Each independently is preferred to be -SR 58 A substituted hydrocarbon group having 1 to 12 carbon atoms, or a hydrocarbon group having 1 to 12 carbon atoms.

[0224] L 31 L 32 L 33 and L 34 Each is preferably -C(O)O- or -OC(O)-.

[0225] R 58 Preferably, it is a hydrocarbon group having 1 to 10 carbon atoms, and more preferably a hydrocarbon group having 1 to 8 carbon atoms.

[0226] The compounds according to embodiments of the present invention can form salts (e.g., pharmaceutically acceptable salts).

[0227] Examples of salts containing basic groups include salts with inorganic acids such as hydrochloric acid, hydrobromic acid, nitric acid, and sulfuric acid; salts with organic carboxylic acids such as formic acid, acetic acid, citric acid, oxalic acid, fumaric acid, maleic acid, succinic acid, malic acid, tartaric acid, aspartic acid, trichloroacetic acid, and trifluoroacetic acid; and salts with sulfonic acids such as methanesulfonic acid, benzenesulfonic acid, p-toluenesulfonic acid, mesitylenesulfonic acid, and naphthalenesulfonic acid.

[0228] Examples of salts with acidic groups include salts with alkali metals such as sodium and potassium; salts with alkaline earth metals such as calcium and magnesium; ammonium salts; and salts with nitrogen-containing organic bases such as trimethylamine, triethylamine, tributylamine, pyridine, N,N-dimethylaniline, N-methylpiperidine, N-methylmorpholine, diethylamine, dicyclohexylamine, procaine, dibenzylamine, N-benzyl-β-phenylethylamine, L-diphenylhydroxymethylamine, and N,N'-dibenzylethylenediamine.

[0229] Among the salts mentioned above, pharmacologically acceptable salts are preferred.

[0230] The lipids represented by formula (VII) and their preparation methods are described in WO2022 / 230964A, the entire teachings of which are incorporated herein by reference.

[0231] Exemplary lipids represented by formula (VII) include: FL-E FL-F FL-G FL-H FL-I .

[0232] From R 51 or R 52The hydrocarbon group having 1 to 21 carbon atoms is preferably an alkyl group having 1 to 21 carbon atoms, an alkenyl group having 2 to 21 carbon atoms, or an alkynyl group having 2 to 21 carbon atoms, more preferably an alkyl group having 1 to 21 carbon atoms or an alkenyl group having 2 to 21 carbon atoms. The alkyl group having 1 to 21 carbon atoms can be straight-chain or branched, and can be chain-like or cyclic. The number of carbon atoms is preferably 3 to 21, and more preferably 5 to 21 carbon atoms. Examples include propyl groups, isopropyl groups, cyclopropyl groups, butyl groups, isobutyl groups, tert-butyl groups, cyclobutyl groups, pentyl groups, cyclopentyl groups, hexyl groups, cyclohexyl groups, heptyl groups, octyl groups, nonyl groups, decyl groups, undecyl groups, dodecyl groups, tridecyl groups, trimethyldodecyl groups (preferably 3,7,11-trimethyldodecyl groups), tetradecyl groups, pentadecyl groups, hexadecyl groups, heptadecanyl groups, and octadecyl groups. The alkenyl groups having 2 to 18 carbon atoms can be straight-chain or branched, and can be chain-like or cyclic. The number of carbon atoms is preferably 3 to 21, and more preferably 5 to 18. Examples include allyl groups, isopentenyl groups, pentenyl groups, hexenyl groups, heptenyl groups, octenyl groups, nonenyl groups (preferably (Z)-2-nonenyl groups or (E)-2-nonenyl groups), decenyl groups, undecenyl groups, dodecenyl groups, dodecadienyl groups, tridecenyl groups (preferably (Z)-twelfthoc-8-enyl groups), tetradecenyl groups (preferably tetradec-9-enyl groups), and pentadecenyl groups (preferably (Z)-tenthoc-8-enyl groups). The alkynyl group having 2 to 21 carbon atoms can be straight-chain or branched, and can be chain-like or cyclic. The number of carbon atoms is preferably 3 to 21, and more preferably 5 to 21. Examples include propynyl groups, butynyl groups, pentyynyl groups, hexynyl groups, heptyynyl groups, octyynyl groups, nonynyl groups, decyynyl groups, undecynyl groups, dodecynyl groups, tetradecynyl groups, pentadecynyl groups, hexadecynyl groups, heptynyl groups, octadecynyl groups, etc. Examples of hydrocarbon groups having 1 to 18 carbon atoms include those examples specifically listed among hydrocarbon groups having 1 to 21 carbon atoms, i.e., hydrocarbon groups having 1 to 18 carbon atoms.

[0233] For cyclic hydrocarbon groups, cycloalkyl groups having 3 to 10 carbon atoms, cycloalkenyl groups having 3 to 10 carbon atoms, cycloalkynyl groups having 3 to 10 carbon atoms, or aryl groups having 6 to 10 carbon atoms are preferred.

[0234] For hydrocarbon groups having 1 to 6 carbon atoms, alkyl groups having 1 to 6 carbon atoms, alkenyl groups having 2 to 6 carbon atoms, or alkynyl groups having 2 to 6 carbon atoms are preferred, and alkyl groups having 1 to 6 carbon atoms or alkenyl groups having 2 to 6 carbon atoms are more preferred. The alkyl groups having 1 to 6 carbon atoms can be straight-chain or branched, and can be chain-like or cyclic. Specific examples include propyl groups, isopropyl groups, cyclopropyl groups, butyl groups, isobutyl groups, tert-butyl groups, cyclobutyl groups, pentyl groups, cyclopentyl groups, and hexyl groups. The alkenyl groups having 2 to 6 carbon atoms can be straight-chain or branched, and can be chain-like or cyclic. Specific examples include allyl, isopentenyl, pentenyl, and hexenyl. The alkynyl groups having 2 to 6 carbon atoms can be straight-chain or branched, and can be chain-like or cyclic. Specific examples include propynyl, butynyl, pentylyl, and hexynyl.

[0235] From R 66 and R 67 The hydrocarbon group having 1 to 10 carbon atoms is preferably an alkyl group having 1 to 10 carbon atoms, an alkenyl group having 2 to 10 carbon atoms, or an alkynyl group having 2 to 10 carbon atoms, and more preferably an alkyl group having 1 to 10 carbon atoms or an alkenyl group having 2 to 10 carbon atoms. The alkyl group having 1 to 10 carbon atoms can be straight-chain or branched, and can be chain-like or cyclic. The number of carbon atoms is preferably 3 to 10, and more preferably 5 to 10 carbon atoms. Specific examples include propyl groups, isopropyl groups, cyclopropyl groups, butyl groups, isobutyl groups, tert-butyl groups, cyclobutyl groups, pentyl groups, cyclopentyl groups, hexyl groups, cyclohexyl groups, heptyl groups, octyl groups, nonyl groups, and decyl groups. The alkenyl group having 2 to 10 carbon atoms can be straight-chain or branched, and can be chain-like or cyclic. The number of carbon atoms is preferably 3 to 10, more preferably 5 to 10. Specific examples include allyl groups, isopentenyl groups, pentenyl groups, hexenyl groups, heptenyl groups, octenyl groups, nonenyl groups (preferably (Z)-2-nonenyl groups or (E)-2-nonenyl groups), and decenyl groups. The alkynyl group having 2 to 10 carbon atoms can be straight-chain or branched, and can be chain-like or cyclic. The number of carbon atoms is preferably 3 to 10, more preferably 5 to 10. Specific examples include propynyl groups, butynyl groups, pentyynyl groups, hexynyl groups, heptenyl groups, octyynyl groups, nonynyl groups, and decynyl groups.

[0236] Examples of ionizable lipids of formula (II) include: MC3; (4-(dimethylamino)butyric acid [(6Z,9Z,28Z,31Z)-heptadecane-6,9,28,31-tetraen-19-yl] ester) WO2010 / 054405 L-319; (9-[4-(dimethylamino)butyryloxy]heptadecanedioic acid bis[(Z)-non-2-enyl] ester) WO2011 / 153493, WO2013 / 086354, WO2013 / 086322 ALC-0315; (6-[6-(2-hexyldecanoyloxy)hexyl-(4-hydroxybutyl)amino]hexyl ester of 2-hexyldecanoic acid)WO2017 / 075331 SM-102; (8-[2-hydroxyethyl-(6-oxo-6-undecyloxyhexyl)amino]octanoic acid heptadecaned-9-ester)WO2017 / 099823 Lipid 5; (8-[(8-heptadecane-9-yloxy-8-oxooctyl)-(2-hydroxyethyl)amino]nonyl octanoate)WO2017 / 099823 Lipid 29; (8-[(8-heptadecane-9-yloxy-8-oxooctyl)-[3-[[2-(methylamino)-3,4-dioxocyclobuten-1-yl]amino]propyl]amino]undecane-3-ester of octanoate) Adv. Funct. Mater.2021, 2106727, DOI: 10.1002 / adfm.202106727 ATX-100; Pentadecan-8-ester of 4-[3-(dimethylamino)propylthiocarbonyl-(4-oxo-4-pentadecan-8-yloxybutyl)amino]butyrate WO2019 / 191780 Lipid A9; (10-[3-(dimethylamino)propyl-nonanoylamino]nonadecanedioic acid bis(2-butyloctyl) ester) WO2017 / 004143 Lp01; ((9Z,12Z)-octadecane-9,12-dienoic acid [2-[3-(diethylamino)propoxycarbonyloxymethyl]-3-(4,4-dioctyloxybutyryloxy)propyl] ester) WO2015 / 09534, WO2020 / 219876 TCL053; ((Z)-Tetradecano-9-enoic acid [2-[4-(dimethylamino)butyryloxymethyl]-3-[(Z)-tetradecano-9-enoyl]oxy-2-[[(Z)-tetradecano-9-enoyl]oxymethyl]propyl] ester) WO 2020 / 032184 TCL065; (3-Pentyloctanoic acid [2-[5-(dimethylamino)pentanoyloxymethyl]-3-(3-pentyloctanoyloxy)-2-(3-pentyloctanoyloxymethyl)propyl] ester) WO 2020 / 032184 Lipid 9; ((Z)-Undecano-5-enoic acid [(6Z,16Z)-12-[6-(dimethylamino)hexanoyloxy]tecosano-6,16-dien-11-yl] ester) WO2021 / 188389 Lipid 19; ((9Z,12Z)-octadec-9,12-dienoic acid [(6Z,16Z)-12-[6-(dimethylamino)hexanoyloxy]teicosto-6,16-dien-11-yl] ester) WO2021 / 188389 and 13-B43; (5-(dimethylamino)valerate [(6Z,16Z)-12-[(Z)-dec-4-enyl]docosa-6,16-dien-11-yl] ester) WO2020 / 219941 .

[0237] Other ionizable lipids applicable to this disclosure include: 306Oi10; (3-[3-[3-[bis[3-(8-methylnonoxy)-3-oxopropyl]amino]propyl-methylamino]propyl-[3-(8-methylnonoxy)-3-oxopropyl]amino]propionic acid 8-methylnonyl ester) 93-O17S; (3-[3-imidazol-1-ylpropyl-[3-oxo-3-(2-tetradecylthioethoxy)propyl]amino]propionic acid 2-tetradecylthioethyl ester) 93-O17O; (2-Tetradecyloxyethyl 3-[3-imidazol-1-ylpropyl-[3-oxo-3-(2-tetradecyloxyethoxy)propyl]amino]propionic acid) 80-O16B; (2-(dodecyldithio)ethyl ester of 3-[3-(dimethylamino)propyl-[3-[2-(dodecyldithio)ethoxy]-3-oxopropyl]amino]propionic acid) CL4H6; ((Z)-octadecyl-9-enoic acid [7-[4-(dipropylamino)butyl]-7-hydroxy-13-[(Z)-octadecyl-9-enoyl]oxy-tetrazyl] ester) YSK05; (1-Methyl-4,4-bis[(9Z,12Z)-octadec-9,12-dienoxy]piperidine) Lipid C24; (3-[4-(4-methylpiperazin-1-yl)butyl-[3-(2-octyldodecyl)-3-oxopropyl]amino]propionate 2-octyldodecyl ester) and 5A2-SC8; 4,10,16-tris(3-(2-((2-methyl-3-(octylthio)propionyl)oxy)ethoxy)-3-oxopropyl)-4,7,10,13,16-pentazanonadecanedioic acid bis(2-((2-methyl-3-(octylthio)propionyl)oxy)ethyl) ester (5 ionizable amino groups + 10 ester groups) .

[0238] Other ionizable lipids applicable to this disclosure include: cKK-E12, MD1; (3,6-bis[4-[bis(2-hydroxydodecyl)amino]butyl]piperazine-2,5-dione) C12-200; (1-[2-[bis(2-hydroxydodecyl)amino]ethyl-[2-[4-[2-[bis(2-hydroxydodecyl)amino]ethyl]piperazine-1-yl]ethyl]amino]dodecane-2-ol) E. Cationic lipids The cationic lipids suitable for the compositions of this disclosure include, for example, a variety of cationic lipids. As used herein, a “cationic lipid” comprises at least one permanently positively charged portion. The permanently positively charged portion may be positively charged at physiological pH, such that the cationic lipid carries a positive charge when delivering a pharmaceutical or therapeutic agent to cells. In some embodiments, the positively charged portion is a quaternary ammonium or quaternary ammonium ion. In some embodiments, the cationic lipid comprises a counterion or otherwise complexes with or interacts with a counterion. Cationic lipids that can be used in this disclosure comprise one or more hydrophobic components and a permanently cationic group. The permanently cationic lipid may contain a group that is positively charged regardless of pH. One type of permanently cationic group that can be used in the permanently cationic lipid is a quaternary ammonium group.

[0239] The cationic lipid component of the lipid nanoparticles of the present invention may account for 1 mol% to 99 mol% of the lipid molecules in the nanoparticles. In other embodiments, the cationic lipid component may account for 5 mol% to 90 mol% of the lipid molecules in the lipid nanoparticles. In some embodiments, the cationic lipid component accounts for 10 mol% to 75 mol%, 10 mol% to 60 mol%, 10 mol% to 50 mol%, or 15 mol% to 40 mol% of the lipid molecules in the lipid nanoparticles.

[0240] In some embodiments, the cationic lipids suitable for use in this disclosure are biodegradable. The cationic lipids suitable for use in this disclosure may have one or more biodegradable groups located in the hydrophobic portion of the cationic lipid (e.g., within a hydrocarbon chain). The inclusion of biodegradable groups in the cationic lipids allows for more rapid metabolism and removal from the body after delivery of the active agent to the target region. Therefore, these cationic lipids may exhibit lower toxicity than similar cationic lipids without biodegradable groups. These cationic lipids can be incorporated into lipid particles to deliver active agents, such as nucleic acids.

[0241] The cationic lipids used in this disclosure may be lipids of formula (IV): in: R 101 and R 102 Each independently constitutes (C8-C)24 )alkyl, (C8-C 24 Alkenyl or any of the above-mentioned groups in a substituted form; R 103 In each case, it is independently (C1-C6)alkyl or substituted (C1-C6)alkyl; R 104 It is a (C1-C6) alkyl or a substituted (C1-C6) alkyl; and X⁻ is a monovalent anion.

[0242] Examples of cationic lipids suitable for use in this disclosure include, but are not limited to: 14:1 EPC (1,2-dimyristoleoyl- sn -glycerol-3-ethylphosphorylcholine), especially as an OTf salt; EDLPC (1,2-dilauryl- sn -glycerol-3-ethylphosphorylcholine), especially as a chloride salt; EDMPC (1,2-dimyristoyl- sn -glycerol-3-ethylphosphorylcholine), especially as a chloride salt; EDPPC (1,2-dipalmitoyl- sn -glycerol-3-ethylphosphorylcholine), especially as a chloride salt; EDSPC (1,2-distearate- sn 3-glycerol-3-ethylphosphorylcholine), particularly as a chloride salt; EDOPC (1,2-dioleoyl- sn -glycerol-3-ethylphosphorylcholine), especially as a chloride salt; and 16:0-18:1 EPC (1-palmitoyl-2-oleoyl- sn 3-glycerol-3-ethylphosphorylcholine, especially as a chloride salt.

[0243] 14:1 EPC EDLPC EDMPC EDPPC EDSPC EDOPC 16:0-18:1 EPC In some embodiments, the cationic lipids suitable for use in this disclosure are cationic lipids of formula (V): in: R 101 and R 102 Each independently constitutes (C8-C) 24 )alkyl, (C8-C 24 Alkenyl or any of the above-mentioned groups in a substituted form; R 103 In each case, it is independently (C1-C6)alkyl or substituted (C1-C6)alkyl; and X⁻ is a monovalent anion.

[0244] Other examples of cationic lipids suitable for use in this disclosure include, but are not limited to: DMTAP (1,2-dimyristoyl-3-trimethylammonium-propane), particularly as a chloride salt; DPTAP (1,2-dipalmitoyl-3-trimethylammonium-propane), particularly as a chloride salt; DSTAP (1,2-distearyl-3-trimethylammonium-propane), particularly as a chloride salt; DOTAP (1,2-dioleoyl-3-trimethylammonium-propane), particularly as a chloride salt; and DORI (N-(2-hydroxyethyl)-N,N-dimethyl-2,3-bis(oleoyloxy)prop-1-ammonium), particularly as a bromide salt.

[0245] DMTAP: DPTAP: DSTAP: DORI: In some embodiments, the cationic lipids suitable for this disclosure are lipids of formula (VI): in: R 101 and R 102 Each independently constitutes (C8-C) 24 )alkyl, (C8-C 24 Alkenyl or any of the above-mentioned groups in a substituted form; R 105 Each is independently a (C1-C6) alkyl group; and X⁻ is a monovalent anion.

[0246] Another example of a cationic lipid suitable for this disclosure is DC-6-14 (O,O'-bis(tetradecanoyl)-N-(α-trimethylammonium acetyl)diethanolamine), particularly in the form of a chloride salt.

[0247] DC-6-14 F. Phospholipids In some embodiments, the compositions of this disclosure comprise phospholipids. As used herein, "phospholipid" refers to a lipid molecule comprising at least one hydrocarbon tail group, a glycerol or sphingosine moiety, and optionally a small organic moiety, preferably selected from amino acids, sugars, choline, or ethanolamine. The phospholipids of this disclosure, such as those containing choline, ethanolamine, or amino acid moieties, may have positively charged groups. However, because of the negative charge present on the lipid molecule, phospholipids as defined herein differ from ionizable lipids or cationic lipids as defined herein. Therefore, the phospholipids of this disclosure are zwitterionic, and the term "phospholipid" may be understood to be synonymous with "zwitterionic phospholipid" as used herein.

[0248] The phospholipid component of the lipid nanoparticles of the present invention may account for 0.1 mol% to 99 mol% of the lipid molecules in the nanoparticles. In other embodiments, the phospholipid component may account for 0.1 mol% to 75 mol% of the lipid molecules in the lipid nanoparticles. In some embodiments, the phospholipid component accounts for 1 mol% to 50 mol%, 1 mol% to 30 mol%, 1 mol% to 15 mol%, or 5 mol% to 10 mol% of the lipid molecules in the lipid nanoparticles.

[0249] Phospholipids applicable to this disclosure include, but are not limited to, DSPC (1,2-distearate- sn -glycerol-3-phosphocholine), DMPC (1,2-dimyristic- sn -glycerol-3-phosphatidylcholine), DPPC (1,2-dipalmitoyl- sn -glycerol-3-phosphatidylcholine), DSPC (1,2-distearate- sn -glycerol-3-phosphatidylcholine), POPC (l-palmitoyl-2-oleoyl- sn -glycerol-3-phosphocholine), DOPC (1,2-dioleoyl- sn -glycerol-3-phosphocholine), DMPE (1,2-dimyristic- sn -glycerol-3-phosphoethanolamine), DPPE (1,2-dipalmitoyl- sn -glycerol-3-phosphoethanolamine), DSPE (1,2-distearate- sn -glycerol-3-phosphoethanolamine), DOPE (1,2-dioleoyl- sn -glycerol-3-phosphoethanolamine), DLoPE (1,2-dilinoleyl- sn -glycerol-3-phosphoethanolamine), D(Phy)PE (1,2-diphydanyl- sn -glycerol-3-phosphoethanolamine), POPE (l-palmitoyl-2-oleoyl- sn -glycerol-3-phosphoethanolamine), 1,2-bis(tetradecyl)- sn -glycerol-3-phosphoethanolamine, 1,2-bis(hexadecyl)- sn -glycerol-3-phosphoethanolamine, 1,2-bis(octadecyl)- sn -Glyceryl-3-phosphoethanolamine, 1,2-diphyllyl- sn -Glyceryl-3-phosphatidylethanolamine, etc. In some embodiments, the phospholipid is phosphatidylcholine. In some embodiments, the phospholipid is phosphatidylethanolamine. In some embodiments, the phospholipid is DSPC.

[0250] G. Sterols In some embodiments, the compositions of this disclosure comprise sterols. As used herein, "sterol" refers to steroids and steroid derivatives, wherein the term "steroid" is a class of compounds having a tetracyclic 17-carbon ring structure, which may further comprise one or more substitutions (including alkyl groups, alkoxy groups, hydroxy groups, oxo groups, acyl groups) or double bonds between two or more carbon atoms. In one aspect, the ring structure of steroids comprises three fused cyclohexyl rings and a fused cyclopentyl ring, such as the structure of cholesterol.

[0251] In some embodiments, the sterols suitable for use in this disclosure are derivatives of cholesterane. As described above, cholesterane derivatives include one or more non-alkyl substitutions in a fused ring system. In some embodiments, the cholesterane or cholesterane derivative is cholesterene or a cholesterene derivative.

[0252] The sterols applicable to this disclosure include, but are not limited to, cholesterol, sitosterol, stigmasterol, fucosterol, spinachsterol, rapeseed sterol, ergosterol, cholesterol ketones, cholesterolenones, coprostanol, cholesterol-2'-hydroxyethyl ether, cholesterol-4'-hydroxybutyl ether, etc. In some embodiments, the sterols of this disclosure are phytosterols. In some embodiments, the sterols of this disclosure are cholesterol.

[0253] The sterol component of the lipid nanoparticles of the present invention may account for 1 mol% to 99 mol% of the lipid molecules in the nanoparticles. In other embodiments, the sterol component may account for 1 mol% to 75 mol% of the lipid molecules in the lipid nanoparticles. In some embodiments, the sterol component accounts for 5 mol% to 75 mol%, 5 mol% to 60 mol%, 10 mol% to 50 mol%, or 15 mol% to 45 mol% of the lipid molecules in the lipid nanoparticles.

[0254] H. PEG-lipids In some embodiments, the compositions of this disclosure comprise PEG-lipids. As used herein, "PEG-lipid" refers to a polyethylene glycol (PEG) polymer linked to one or more hydrocarbon chains of 1-30 carbon atoms in length. In some embodiments, a PEG-lipid is a compound in which one or more hydrocarbon chains of 1-30 carbon atoms in length are linked to a linking group, which in turn is linked to a PEG chain. In some embodiments, the PEG-lipid is a diglyceride that further comprises a PEG chain linked to a glycerol group.

[0255] The PEG lipid component of the lipid nanoparticles of the present invention may account for 0.1 mol% to 99 mol% of the lipid molecules in the nanoparticles. In other embodiments, the PEG lipid component may account for 0.1 mol% to 75 mol% of the lipid molecules in the lipid nanoparticles. In some embodiments, the PEG lipid component accounts for 0.1 mol% to 25 mol%, 0.1 mol% to 10 mol%, 0.1 mol% to 5 mol%, or 0.1 mol% to 3 mol% of the lipid molecules in the lipid nanoparticles.

[0256] Some non-limiting examples of PEG lipids include PEG-modified phosphatidylethanolamine or phosphatidic acid, PEG-ceramide conjugates, PEG-modified dialkylamines, PEG-modified 1,2-diacyloxypropyl-3-amine, PEG-modified diacylglycerols, and dialkylglycerols. In some embodiments, the PEG lipids suitable for use in this disclosure are PEG-modified distearate phosphatidylethanolamine or PEG-modified dimyristoyl- sn- Glycerol. In some embodiments, the PEG modification is determined by the molecular weight of the PEG component of the lipid. In some embodiments, the PEG modification has a molecular weight of about 100 to about 15,000. In some embodiments, the molecular weight is about 200 to about 500, about 400 to about 5,000, about 500 to about 3,000, or about 1,200 to about 3,000. The molecular weight of the PEG modification is about 100, 200, 400, 500, 600, 800, 1000, 1250, 1500, 1750, 2000, 2250, 2500, 2750, 3000, 3500, 4000, 4500, 5000, 6000, 7000, 8000, 9000, 10000, 12500 to about 15000. In some embodiments, the PEG modification is determined by the number of repeating subunits in the polymer chain. Some non-limiting examples of lipids that may be used in this application are taught by U.S. Patent 5,820,873, WO 2010 / 141069 or U.S. Patent 8,450,298, which are incorporated herein by reference.

[0257] The PEG-lipids applicable to this disclosure include, but are not limited to, DMG-mPEG2000 (1,2-dimyristoyl-rac-glycerol-3-methoxy polyethylene glycol-2000), DPG-mPEG2000 (1,2-dispalmitoyl-rac-glycerol-3-methoxy polyethylene glycol-2000), and DSG-mPEG2000 (1,2-distearateyl-rac-glycerol-3-methoxy polyethylene glycol-2000). Although PEG modification can involve the non-region-specific addition of PEG groups and varying numbers of repeating monomers, resulting in listed molecular weights representing averages, typical structures represented by DMG-mPEG2000 include, for example: .

[0258] I. Application The compositions disclosed herein comprise a plurality of lipid particles (e.g., lipid nanoparticles, such as any lipid nanoparticles described herein). In some embodiments, the compositions are pharmaceutical compositions. In some embodiments, the compositions further comprise a pharmaceutically acceptable carrier.

[0259] "Pharmaceutically acceptable carrier" refers to a non-toxic carrier or excipient that does not impair the pharmacological activity of the drug formulation with which it is prepared and is non-toxic when administered at a dose sufficient to deliver a therapeutic amount of the drug. Pharmaceutically acceptable carriers that may be used in the compositions described herein include, but are not limited to, ion exchangers, alumina, aluminum stearate, lecithin, serum proteins such as human serum albumin, buffering substances such as phosphates, glycine, sorbic acid, potassium sorbate, mixtures of partial glycerides of saturated vegetable fatty acids, water, salts or electrolytes such as protamine sulfate, disodium hydrogen phosphate, potassium hydrogen phosphate, sodium chloride, zinc salts, colloidal silica, magnesium trisilicate, polyvinylpyrrolidone, cellulose-based substances, polyethylene glycol, sodium carboxymethyl cellulose, polyacrylates, waxes, polyethylene-polyoxypropylene blocks, polymers, polyethylene glycol, and lanolin.

[0260] The compositions described herein can be administered parenterally (including subcutaneously, intramuscularly, intravenously, and intradermally), by inhalation spray, orally, topically, rectally, nasally, orally, vaginally, or through an implanted reservoir. As used herein, the terms “parenteral” and “extracorporeal” include subcutaneous, intradermal, intravenous, intramuscular, intraocular, intravitreal, intraarticular, intraarticular, intrasynovial, intrasternal, intrasheath, intralesional, intrahepatic, intraperitoneal, intralesional, and intracranial injection or infusion techniques. In some aspects, the compositions described herein can be administered intravenously and / or intraperitoneally. In some aspects, the compositions described herein can be administered orally. In some aspects, the compositions described herein can be administered subcutaneously. Preferably, the compositions described herein are administered orally, subcutaneously, intraperitoneally, or intravenously. More preferably, the compositions described herein are administered intravenously.

[0261] The compositions described herein can also be administered subcutaneously, intraperitoneally, or intravenously, for example, as a sterile injectable formulation, such as a sterile injectable aqueous or oily suspension. The suspension can be formulated using suitable dispersants or wetting agents (e.g., Tween 80) and suspending agents according to techniques known in the art. The sterile injectable formulation can also be a sterile injectable solution or suspension in a non-toxic, parenteral-acceptable diluent or solvent, such as a solution in 1,3-butanediol. Acceptable carriers and solvents that can be used are mannitol, dextran, water, phosphate-buffered saline, Ringer's solution, lactated Ringer's solution, and isotonic sodium chloride solution. Common surfactants typically used to prepare pharmaceutically acceptable solid, liquid, or other dosage forms, such as Tween or Span and / or other similar emulsifiers or bioavailability enhancers, can also be used for formulation purposes.

[0262] As directed, administration may be local or systemic. In some embodiments (e.g., of the compositions disclosed herein), administration is oral. In some embodiments (e.g., of the compositions disclosed herein), administration is intravenous. Preferred administration methods may vary depending on the specific composition or agent to be delivered.

[0263] J. Usage Method The lipid compositions disclosed herein can be used to deliver pharmaceutical agents to a variety of tissues and organs within an individual. In some aspects, this disclosure describes a method of delivering a pharmaceutical agent to one or more of the brain, heart, muscle, and kidneys of an individual in need, the method comprising systemically administering to the individual a composition comprising a plurality of lipid nanoparticles, wherein the lipid nanoparticles comprise the pharmaceutical agent, ionizable lipids, and fully saturated cationic lipids. In some embodiments, the pharmaceutical agent is delivered to the brain. In some embodiments, the pharmaceutical agent is delivered to the muscle. In some embodiments, the pharmaceutical agent is delivered to the heart. In some embodiments, the pharmaceutical agent is delivered to the kidney. In some embodiments, the pharmaceutical agent is delivered to both the heart and muscle. In some embodiments, the pharmaceutical agent is delivered to both the heart and brain. In some embodiments, the pharmaceutical agent is delivered to both the heart and kidney. In some embodiments, the pharmaceutical agent is delivered to both the brain and muscle. In some embodiments, the pharmaceutical agent is delivered to both the brain and kidney. In some embodiments, the pharmaceutical agent is delivered to both muscle and kidney. In some embodiments, the pharmaceutical agent is delivered to both muscle, kidney, and heart. In some embodiments, the agent is delivered to the muscles, kidneys, and brain. In some embodiments, the agent is delivered to the brain, kidneys, and heart. In some embodiments, the agent is delivered to the muscles, brain, and heart.

[0264] In some aspects, this disclosure describes methods of delivering a pharmaceutical agent to an individual in need, the methods comprising administering a composition of the present disclosure to the individual. In some aspects, this disclosure describes methods of treating an individual suffering from a disease, disorder, or condition that is beneficially treatable by a pharmaceutical agent, the methods comprising administering a therapeutically effective amount of a composition of the present disclosure to the individual. In some embodiments, the ionizable lipid in the administered composition is an ionizable lipid of formula (I). In some embodiments, the ionizable lipid in the administered composition is an ionizable lipid of formula (II). In some embodiments, the ionizable lipid in the administered composition is an ionizable lipid of formula (III). In some embodiments, the ionizable lipid in the administered composition is an ionizable lipid of formula (VII). In some embodiments, the ionizable lipid in the administered composition is an ionizable lipid of formula (VIII). In some embodiments, the ionizable lipid in the administered composition is an ionizable lipid of formula (IX). In some embodiments, the ionizable lipid in the administered composition is an ionizable lipid of formula (X). In some embodiments, the cationic lipid in the applied composition is a cationic lipid of formula (IV). In some embodiments, the cationic lipid in the applied composition is a cationic lipid of formula (V). In some embodiments, the cationic lipid in the applied composition is a cationic lipid of formula (VI).

[0265] Typically, the compositions of this disclosure are applied once every 1 to 30 days. In some embodiments, the application is once every 30 days. In some embodiments, the application is once every 25 days. In some embodiments, the application is once every 20 days. In some embodiments, the application is once every 10 days. In some embodiments, the application is once every 9 days. In some embodiments, the application is once every 8 days. In some embodiments, the application is once every 7 days. In some embodiments, the application is once every 6 days. In some embodiments, the application is once every 5 days. In some embodiments, the application is once every 4 days. In some embodiments, the application is once every 3 days. In some embodiments, the application is once every 2 days. In some aspects, (e.g., of the compositions of this disclosure) the application is QD or BID (e.g., QD). In some aspects, (e.g., of the compositions of this disclosure) the application is daily.

[0266] In some aspects, the compositions described herein also comprise one or more other pharmaceutical agents, for example, for use in combination with the first pharmaceutical agent. Some embodiments provide combinations comprising pharmaceutical agents in addition to one or more therapeutic agents (e.g., drug combinations). Such combinations are particularly useful, for example, when the first pharmaceutical agent and one or more other therapeutic agents need to be administered individually. In the combinations provided herein, two or more pharmaceutical agents may be administered via the same route of administration or via different routes of administration. In some embodiments, the other pharmaceutical agent is a therapeutic agent. In some embodiments, the other pharmaceutical agent is a diagnostic agent. In some embodiments, the pharmaceutical agent is an imaging agent. In some aspects, the other pharmaceutical agent is a small molecule. In some aspects, the other pharmaceutical agent is a peptide. In some aspects, the other pharmaceutical agent is a protein. In some aspects, the other pharmaceutical agent is an antibody or antibody fragment.

[0267] In some embodiments of this disclosure, the pharmaceutical agent in the composition is a therapeutic agent for treating a disease, disorder, or condition. The compositions of this disclosure may also be administered in combination with one or more other therapies to treat a disease, disorder, or condition. When administered in combination with such other therapies, the compounds of this disclosure may be administered before, after, or simultaneously with the other therapies (e.g., other therapeutic agents). When administered concurrently (e.g., simultaneously), the compositions of this disclosure and another therapy may be in separate formulations or in the same formulation. Furthermore, the other therapy may be in a solvent of the formulation or bound to lipid nanoparticles, including by covalent or non-covalent bonding to a surface, embedded in a membrane, or encapsulated within nanoparticles. Alternatively, the compositions of this disclosure and another therapy may be administered as separate formulations at substantially the same time or in different time sequences. When the compositions of this disclosure and other therapies (e.g., therapeutic agents) are administered as separate formulations or compositions, the compositions of this disclosure and other therapies may be administered via the same route of administration or via different routes of administration. A skilled clinician can determine the appropriate timing of administration for each therapy used in combination (e.g., a timing sufficient to allow for overlap in the efficacy of the therapies). Typically, combination therapy provides the beneficial effects of a combination of therapeutic agents in treating the diseases, conditions, or disorders described in this article.

[0268] The compositions or other therapeutic agents disclosed herein may be administered at doses of about 0.001 mg / kg to about 100 mg / kg body weight, or alternatively at doses of about 1 mg / dose to about 5000 mg / dose, or as required by a particular pharmaceutical agent. For example, suitable doses may be about 0.001 mg / kg to about 100 mg / kg body weight, about 0.01 mg / kg to about 100 mg / kg body weight, about 0.01 mg / kg to about 10 mg / kg body weight per treatment. In some aspects, suitable doses are about 0.1 mg / kg to about 10 mg / kg body weight per treatment, such as about 0.1 mg / kg to about 5 mg / kg body weight, about 0.1 mg / kg to about 2.5 mg / kg body weight, or about 0.1 mg / kg to about 1.0 mg / kg body weight. Suitable doses may be about 0.001 mg / dose to about 100 mg / dose, about 0.01 mg / dose to about 100 mg / dose, about 0.01 mg / dose to about 50 mg / dose, about 0.1 mg / dose to about 10 mg / dose, about 0.05 mg / dose to about 50 mg / dose, about 0.1 mg / dose to about 100 mg / dose, about 1 mg / dose to about 7500 mg / dose, about 1 mg / dose to about 5000 mg / dose, about 10 mg / dose to about 2500 mg / dose, or about 100 mg / dose to about 1000 mg / dose.

[0269] Doses lower or higher than those described above may be required. The specific dosage and treatment regimen for any particular patient depends on a variety of factors, such as the activity of the specific drug used, age, weight, general health status, sex, diet, timing of administration, frequency of administration, rate of excretion, drug combination, severity and duration of the disease, symptoms or conditions, individual susceptibility to the disease, symptoms or conditions, and the judgment of the treating physician. Determining the dosage of a specific drug, individual, and disease, disorder, or condition is within the capabilities of a person skilled in the art.

[0270] K. Example Materials and methods: All chemicals obtained from commercial sources are stored according to the manufacturer's instructions and are ready for use without further purification. Lipids represented by formula (I) or (III) or (VII) and methods for their preparation are described in US2021 / 0085604 and US2022 / 0273817. The entire contents of each disclosure are incorporated herein by reference.

[0271] siRNA: One of the most common evaluation systems for extrahepatic delivery is bioluminescence imaging (BLI) using firefly luciferase (FLuc) mRNA. After administration of a non-toxic, stable substrate, FLuc emits light at tissue-penetrating wavelengths, which can be imaged in vivo and ex vivo, allowing for the identification of tissues where mRNA has been successfully delivered and translated. While these BLI techniques are simple and convenient, quantification of bioluminescence from bulk tissues may overlook RNA delivery to underrepresented cell types within the tissue. This is particularly true for endothelial cells. For example, the lungs and liver contain 15%–30% endothelial cells, while the kidneys and brain contain less than 5%, resulting in less bioluminescence even with efficient FLuc mRNA delivery to endothelial cells. In principle, mRNA encoding fluorescent proteins can facilitate cell-type level analysis using microscopy and flow cytometry. However, currently commercially available GFP and tdTomato mRNAs have been reported to fail to induce sufficient protein expression to visualize beyond background fluorescence in vivo. To address this sensitivity issue, an evaluation system was developed in which Cre recombinase mRNA was administered to genetically engineered mice (Ai14). Transcription of the tdTomato protein, driven by the CAG promoter, was inhibited by a STOP box flanked by loxPs, thus achieving highly sensitive evaluation. However, the Ai14 / Cre mRNA model is an "on / off" binary system; successful transfection of one Cre mRNA results in the same tdTomato expression as many Cre mRNAs, and RNA delivery efficiency (input) is disproportionate to fluorescence intensity and protein expression (output).

[0272] Conversely, siRNA is a useful therapeutic modality and a powerful tool for in vivo screening of LNPs. For example, early efforts to enhance the potency of ionizable lipids were supported by screening systems using siRNA targeting factor VII (FVII), a clotting factor secreted by hepatocytes. Because FVII expression is strictly limited to hepatocytes, the efficiency of RNA delivery to hepatocytes can be readily assessed by quantifying serum FVII protein levels. Regarding endothelial cells, siRNA is used to target genes specifically expressed in the endothelium, such as Icam2, Tie2, and... Cdh5 It can be effective.

[0273] cadherin ( Cdh5 ), also known as vascular endothelial cadherin (VE-cadherin), is a useful target gene for assessing endothelial cell siRNA delivery because... Cdh5 Expression is strictly limited to endothelial cells. The amount retained in tissues after siCdh5 administration was quantified. Cdh5 mRNA can be used to assess the efficiency of RNA delivery to endothelial cells.

[0274] The following custom siRNAs were prepared by Horizon. siRNAs targeting mouse VECadherin (siVEcad, siCdh5): Justice: 5'-mCmCAAAAGAGAGAmCmUGGAmUmUdTsdT-3' (SEQ ID NO:1) Antonym: 5'-AAUCmCAGUCUCUCUUUUGGdTsdT-3' (SEQ ID NO: 2) abbreviation Adenosine-3'-phosphate C-cytidine-3'-phosphate G-guanosine-3'-phosphate Uruidine-3'-phosphate mA 2'-O-methyladenosine-3'-phosphate mC 2'-O-methylcytidine-3'-phosphate mG 2'-O-methylguanosine-3'-phosphate mU 2'-O-methyluridine-3'-phosphate dT 2'-deoxythymidine-3'-phosphate sdT 2'-deoxythymidine-5'-phosphate-thiophosphate Lipid Nanoparticle Formulation: Lipid nanoparticles were synthesized using the previously described microfluidic chip device (Chen, D., J. Am. Chem. Soc. (2012)). This facilitated the rapid discovery of effective siRNA-containing lipid nanoparticles achievable through controlled microfluidic agents. Briefly, a lipid-containing ethanol phase was mixed with an siRNA-containing aqueous phase via microfluidic channels in a PDMS (polydimethylsiloxane) chip. The siRNA was diluted in 10 mM citrate buffer (pH 3.0) (aqueous phase), while an appropriate amount of lipid was co-dissolved in 200 standard strength ethanol (ethanol phase). The ethanol and aqueous phases were mixed together using a syringe pump at a 1:3 volume ratio (total flow rate 1.2 mL / min).

[0275] The obtained LNP was dialyzed overnight against phosphate-buffered saline (PBS) in a 20 kDa molecular weight cutoff (MWCO) box at 4°C or room temperature and stored at 4°C until use. In some experiments, the LNP solution was further concentrated using a 100 kDa MWCO Amiconultra centrifuge filter (Millipore Sigma). For animal experiments, the LNP solution was sterilized by passing it through a 0.22 μm filter (Millipore Sigma).

[0276] Particle size and zeta potential determination: The diameter (z-mean) and polydispersity index (PDI) of LNPs were determined using dynamic light scattering (Zetasizer Nano ZS (Malvern Instruments)). The zeta potential was determined in 0.1X PBS solution using the same instrument.

[0277] Quantification of siRNA Concentration and Encapsulation: To quantify siRNA concentration and determine RNA encapsulation efficiency, the modified Quant-iT RiboGreen RNA assay (Thermo Fisher) was used as previously described (Walsh C. et al., Methods Mol Biol. 2014;1141:109-20). This assay measures the amount of RNA in samples with intact LNPs to determine the amount of unencapsulated RNA, and measures the amount of RNA in LNP samples disrupted by Triton X-100 to quantify total RNA. Briefly, 0.5 μL of LNP of known concentration or serially diluted siRNA was diluted in black 96-well plates with or without 2% Triton X-100 (Sigma-Aldrich) in a final volume of 100 μL of TE buffer (10 mM tris-HCl and 20 mM EDTA). The plates were incubated at 37°C with shaking at 350 rpm for 15 minutes. After incubation, add 99.5 μl of TE buffer and 0.5 μl of RiboGreen reagent to each well. Incubate the plate at 37°C with shaking at 350 rpm for 2 minutes. Measure RiboGreen fluorescence using a ELISA reader (Tecan) (excitation wavelength 485 nm, emission wavelength 528 nm). RNA encapsulation efficiency (EE%) was determined using the following formula: pKa determination by TNS: The apparent pKa values ​​of lipid nanoparticles were determined using the 2-(p-toluidine)naphthalene-6-sulfonic acid (TNS) assay as described above (Heyes J. et al., Journal of Controlled Release 107 (2005) 276–287). Briefly, buffers ranging from pH 3.0 to 9.0 in increments of 0.5 were prepared by mixing 20 mM sodium phosphate buffer, 20 mM sodium citrate buffer, 20 mM sodium borate buffer, and 150 mM NaCl. 300 μM TNS solution was added to the above pH buffers to a final concentration of 6 μM. 46 μL of each pH buffer containing 6 μM TNS was added to each black 384-well plate. Finally, 4 μL of LNP solution containing 500 ng / μL ionizable lipids was added to each well (quadruple). Fluorescence intensity was measured using a Tecan microplate reader at excitation at 320 nm and emission at 465 nm. The obtained pH-fluorescence S-shaped curves were evaluated using Prism (Graphpad) curve fitting analysis, and the apparent pKa values ​​were obtained from the inflection points of the S-shaped curves.

[0278] Animal Studies: All animal studies were approved by the MIT Institutional Animal Care and Use Committee (CAC) and were consistent with applicable local, state, and federal regulations. All experimental procedures were conducted under the guidance of the Division of Comparative Medicine at MIT, in accordance with ethical compliance and approval. Female C57BL / 6 mice (6 weeks old) were obtained from the Jackson Laboratory, placed in the MIT animal facility, and acclimatized for at least 3 days prior to the start of the studies.

[0279] For intravenous administration, after gently heating the animal with a heat lamp, siRNA-lipid nanoparticles diluted in PBS were injected via the tail vein using a 29 g, 3 / 10 cc insulin syringe (BD Biosciences). For intramuscular administration, siRNA-lipid nanoparticles diluted in PBS were injected via the tail vein using a 29 g, 3 / 10 cc insulin syringe (BD Biosciences).

[0280] 48-72 hours after injection, organs or tissues, including heart, liver, spleen, lung, kidney, muscle (quadriceps) and brain, are collected, soaked in RNAlater solution (Thermo Fisher) at 4°C for 12-48 hours, and stored at -20°C after RNAlater removal.

[0281] Tissue mRNA quantification by RT-qPCR: Total RNA was isolated from tissues using TRIzol (Thermo Fisher) and Direct-zol-96 MagBead (Zymo Research) or Quick-RNA MagBead (Zymo Research). Briefly, tissue perforations were placed in 96-well deep-well plates with 4 mm stainless steel beads and lysed using GenoGrinder2010 with 350 μL TRIzol or 250 μL DNA / RNA Shield (Zymo Research). Total RNA was further purified using Direct-zol-96 MagBead (Zymo Research) or Quick-RNA MagBead (Zymo Research) according to the manufacturer's protocol.

[0282] cDNA was synthesized using the ABI High-Capacity cDNA Reverse Transcription Kit (Applied Biosystems #4368814) according to the manufacturer's instructions. In short, for each reaction, 5 μl of a master mix containing 1 μl of 10X buffer, 0.4 μl of 25X dNTPs, 1 μl of 10X random primers, 0.5 μl of reverse transcriptase, and 2.1 μl of water was added to 5 μl of total RNA solution isolated using the above protocol. The plate was sealed, mixed, and incubated on a thermal cycler at 25°C for 10 minutes, then at 37°C for 2 hours, and finally at 85°C for 5 minutes.

[0283] Using the Luna® Universal Probe RT-qPCR Kit (NEB) and Taqman probes— Cdh5 (Mm00486938_m1) Gusb Gene expression was analyzed in qPCR using either (Mm01197698_m1) or B2m (Mm00437762_m1). Samples were amplified using a LightCycler 480 qPCR machine (Roche). Cdh5 Expressing relative B2m or Gusb Normalize.

[0284] Example 1: Comparison of 20% EDPOC LNP and conventional liver-targeted LNP To explore the potential of cationic lipids for extrahepatic RNA delivery, DSPC was first replaced with EDOPC in a standard hepatocyte-targeted LNP formulation. The resulting LNP was then compared with that targeting… Cdh5 The siRNA was formulated to assess gene silencing in endothelial cells in vivo. Formulation details are shown in Table 1. The physicochemical properties of LNPs are shown in Table 2. Results are presented in... Figure 2A-2G These results indicate that replacing zwitterionic phospholipids in liver-targeting LNPs with cationic lipids can alter the tissue and cellular tropism of LNPs.

[0285] Table 1: Formulations

[0286] Table 2: Physicochemical Properties

[0287] Example 2: Comparison of 20% EDOPC LNP and 50% DOTAP LNP Although there have been reports of achieving lung RNA delivery by incorporating 33%–50% DOTAP into liver-targeting LNPs (Q. Cheng, Nat. Nanotechnol., 2020., KJ Kauffman, Mol. Ther. - Nucleic Acids 2018., RJ Dorkin, MIT Thesis, 2016), the use of LNPs to deliver RNA to endothelial cells in organs other than the liver, spleen, and lungs has not been reported. To explore this possibility, LNPs were formulated using 20% ​​EDOPC and compared with previously reported LNPs containing 50% DOTAP. Formulation details are shown in Table 3. The physicochemical properties of the LNPs are shown in Table 4. Results are presented in… Figures 3A-3E middle.

[0288] Table 3: Formulations

[0289] Table 4: Physicochemical Properties

[0290] Example 3: Screening of cationic lipids at 20 mol% To elucidate the relationship between the chemical structure of cationic lipids and their efficiency in delivering RNA outside the liver, spleen, and lungs, a variety of biodegradable cationic lipids were evaluated. Three different classes of cationic lipids were prepared: class 1 consisted of cationic phospholipids, class 2 consisted of trimethylammonium propane (TAP) lipids and DORI, an analog of DOTAP with an ethanolamine head group, and class 3 consisted of DC-6-14. These lipids all contained a hydrophilic quaternary ammonium head group and a biodegradable ester bond located at their hydrophobic tail, but differed in their tail length, saturation, and the connection structure between their head and tail. LNPs of each cationic lipid in 20% concentration were formulated and characterized. All cationic lipids showed pKa values ​​of 6–7 and zeta potentials ranging from near zero to slightly positive. Formulation details are shown in Table 5. The physicochemical properties of the LNPs are shown in Table 6. Results are presented in... Figure 4A-4G LNPs containing EDSPC and DMTAP were excluded from in vivo evaluation due to their large particle size (>150 nm) and inability to be sterilized by filtration.

[0291] Then by using 0.5 mg kg -1Intravenous administration of siCdh5 at various doses was used to evaluate RNA delivery to endothelial cells in multiple organs. While all cationic lipids showed comparable levels of endothelial gene silencing in the liver and spleen, differences in tail structure significantly impacted delivery efficiency in other organs. For example, in terms of unsaturation, compared to C14:1 EPC, C14:0 EPC LNP delivered siRNA more efficiently to endothelial cells in organs such as the lungs, heart, brain, and skeletal muscle. On the other hand, compared to DSTAP (18:0 TAP) LNP and DOTAP (18:1 TAP) LNP, DOTAP LNP showed more efficient endothelial cell delivery in organs such as the kidneys, heart, brain, and skeletal muscle. Tail chain length also showed a significant effect on endothelial delivery efficiency. For example, although DSTAP (18:0 TAP) and DPTAP (16:0 TAP) differed by only two carbon atoms in their tails, DPTAP showed significantly more efficient endothelial delivery.

[0292] Similarly, although EDLPC (12:0 EPC) and EDMPC (14:0 EPC) differ by only two carbon atoms, EDMPC exhibits more efficient endothelial delivery. Furthermore, when comparing EDPPC (16:0 EPC) and EDOPC (18:1 EPC) with 16:0–18:1 EPC (which has an asymmetric tail structure and can be considered an intermediate structure between EDPPC and EDOPC), 16:0–18:1 EPC does not exhibit intermediate properties between EDPPC and EDOPC, showing similar or slightly lower delivery efficiency than EDOPC.

[0293] These results indicate that, in addition to the positive charge in the head, the tail structure of cationic lipids also significantly contributes to their endothelial delivery efficiency. In particular, cationic lipids with symmetrical tails of saturated C14 and saturated C16 (e.g., EDPPC, DPTAP, and DC-6-14) exhibit the most efficient delivery to endothelial cells.

[0294] Table 5: Formulations

[0295] Table 6: Physicochemical Properties

[0296] Example 4: Advantages of EDPPC and DPTAP compared to EDMPC and DOTAP To further confirm the superiority of EDPPC over EDMPC and DPTAP over DOTAP, these cationic lipids were formulated with MC3 and MD-1, and their in vivo endothelial RNA delivery efficiency was evaluated. The encapsulated... Cdh5 The siRNA LNP was administered intravenously to mice at 0.5 mg / kg, and organs were harvested 72 hours post-injection. Total RNA was isolated and purified from the organs, relative to housekeeping genes. Gusb Quantitative Cdh5 mRNA. Formulation details are shown in Table 7. Physicochemical properties of LNPs are shown in Table 8. Results are presented in... Figure 5A-5G and Figure 6A-6G middle.

[0297] Table 7: Formulations

[0298] Table 8: Physicochemical Properties

[0299] Example 5: Combination of ionizable lipids and cationic lipids To further confirm the necessity of the combination of ionizable and cationic lipids for endothelial RNA delivery, LNPs without ionizable lipids were prepared and compared with LNPs containing both ionizable and cationic lipids. The target LNPs were then encapsulated with... Cdh5 The siRNA LNP was administered intravenously to mice at 0.3 mg / kg, and organs were harvested 72 hours post-injection. Total RNA was isolated and purified from the organs, relative to housekeeping genes. Gusb Quantitative Cdh5 mRNA. Formulation details are shown in Table 9. Physicochemical properties of LNPs are shown in Table 10. Results are presented in... Figures 7A-7D middle.

[0300] Table 9: Formulations

[0301] Table 10: Physicochemical Properties

[0302] Example 6: Universality of ionizable lipids (20% EDPPC) To explore the universality of these cationic lipids for extrahepatic delivery, various ionizable lipids were formulated into LNPs containing 20% ​​EDPPC, 20% DPTAP, and 20% DC-6-14. First, LNPs containing 20% ​​EDPPC and each containing 15 different ionizable lipids were prepared.

[0303] Surprisingly, with a single administration of 0.5 mg / kg siCdh5, all LNPs showed efficacy in endothelial cells across all tested organs. Cdh5 Gene silencing. Formulation details are shown in Table 11. Physicochemical properties of LNPs are shown in Table 12. Results are presented in... Figure 8A-8G , Figure 9A-9G and Figure 10A-10G middle.

[0304] Table 11: Formulations

[0305] Table 12: Physicochemical Properties

[0306] Example 7: Universality of ionizable lipids (20% DPTAP) To further explore the universality of these cationic lipids for extrahepatic delivery, various ionizable lipids were formulated into LNPs containing 20% ​​EDPPC, 20% DPTAP, and 20% DC-6-14. LNPs containing 20% ​​DPTAP and each of 11 different ionizable lipids were prepared. Surprisingly, all LNPs showed efficacy in endothelial cells of all test organs after a single administration of 0.5 mg / kg siCdh5. Cdh5 Gene silencing. Formulation details are shown in Table 13. Physicochemical properties of LNPs are shown in Table 14. Results are presented in... Figure 11A-11G and Figure 12A-12G middle.

[0307] Table 13: Formulations

[0308] Table 14: Physicochemical Properties

[0309] Example 8: Universality of ionizable lipids (20% DC-6-14) To further investigate the universality of these cationic lipids for extrahepatic delivery, various ionizable lipids were formulated into LNPs containing 20% ​​EDPPC, 20% DPTAP, and 20% DC-6-14. Finally, the universality of DC-6-14 was further investigated. LNPs containing 20% ​​DC-6-14 and each of 13 different ionizable lipids were further examined. Surprisingly, all LNPs showed efficacy in endothelial cells of all tested organs after a single administration of 0.5 mg / kg siCdh5. Cdh5 Gene silencing. Formulation details are shown in Table 15. Physicochemical properties of LNPs are shown in Table 16. Results are presented in... Figure 13A-13G and Figure 14A-14G middle.

[0310] Table 15: Formulations

[0311] Table 16: Physicochemical Properties

[0312] Example 9: Optimization of EDPPC ratio (0-50 mol%) To optimize the ratio of cationic lipids for extrahepatic delivery, a series of LNPs were prepared by systematically increasing the percentage of cationic lipids identified in the screening. The obtained LNPs were tested in vivo with a single 0.5 mg / kg siCdh5 administration. LNPs containing 10%–50% EDDPC showed efficacy in the liver, spleen, kidney, lung, and brain. Cdh5 Gene silencing was observed, with LNPs containing 20%-30% EDPPC showing the most effective gene silencing in the heart and skeletal muscle. Formulation details are shown in Table 17. The physicochemical properties of LNPs are shown in Table 18. Results are presented in… Figure 15A-15G middle.

[0313] Table 17: Formulations

[0314] Table 18: Physicochemical Properties

[0315] Example 10: Optimization of DPTAP ratio (0-50 mol%) Next, a series of LNPs containing 5%–50% DPTAP were prepared and tested in vivo with a single dose of 0.5 mg / kg siCdh5. LNPs containing 10%–50% DPTAP showed efficacy in the liver, spleen, kidney, lung, heart, and skeletal muscle. Cdh5 Gene silencing. Formulation details are shown in Table 19. Physicochemical properties of LNPs are shown in Table 20. Results are presented in... Figure 16A-16G middle.

[0316] Table 19: Formulations

[0317] Table 20: Physicochemical Properties

[0318] Example 11: Optimization of DC-6-14 ratio (0-50 mol%) Next, a series of LNPs containing 5%–50% DC-6-14 were prepared and tested in vivo with a single dose of 0.5 mg / kg siCdh5. LNPs containing 10%–50% DPTAP showed efficacy in the liver, spleen, kidney, lung, heart, and skeletal muscle. Cdh5 Gene silencing. Formulation details are shown in Table 21. Physicochemical properties of LNPs are shown in Table 22. Results are presented in... Figure 17A-17G middle.

[0319] Table 21: Formulations

[0320] Table 22: Physicochemical Properties

[0321] Example 12: 50% DOTAP with lipids of formula I or III To investigate whether liver-targeted lung-targeting lymphocytes (LNPs) containing 50% DOTAP in lipids comprising formula I or III are redirected to the lungs, five LNPs with different ionizable lipids (MC3 as a positive control) and one LNP without ionizable lipids as a negative control were prepared. Their in vivo lung endothelial delivery efficiency was tested using a single intravenous administration of 0.4 mg / kg siCdh5. Lung tissue was harvested 3 days after administration, and quantification was performed by RT-qPCR. Cdh5 mRNA. Except for 50% DOTAP, all LNPs containing ionizable lipids selected from FL-A, FL-B, FL-C, and FL-D showed 80%-90%. Cdh5 Gene silencing indicates that these novel formulations can be used to deliver RNA to the lungs. Formulation details are shown in Table 23. The physicochemical properties of LNPs are shown in Table 24. Results are presented in... Figure 18 middle.

[0322] Table 23: Formulations

[0323] Table 24: Physicochemical Properties

[0324] Example 13: 50% DOTAP with different PEG lipids To investigate whether the alkyl chain length of PEG-lipids affects the tissue tropism of LNPs containing 50% DOTAP, three LNPs with different PEG lipids were prepared and their endothelial delivery efficiency was tested in vivo using a single intravenous administration of 1.0 mg / kg siCdh5. Tissue samples were obtained 3 days after administration, and quantification was performed by RT-qPCR. Cdh5mRNA. All the LNPs mentioned showed comparable results in all collected tissues. Cdh5 Gene silencing. Formulation details are shown in Table 25. Physicochemical properties of LNPs are shown in Table 26. Results are presented in... Figures 19A-19E middle.

[0325] Table 25: Formulations

[0326] Table 26: Physicochemical Properties

[0327] Example 14: 20% EDOPC with different ionizable lipids To explore the general applicability of EDOPC for extrahepatic delivery, various ionizable lipids were formulated with 20% EDOPC. LNPs containing five different ionizable lipids were prepared, and their endothelial delivery efficiency was tested in vivo using a single intravenous administration of 0.5 mg / kg siCdh5. Three days after administration, tissue samples were obtained, and quantification was performed by RT-qPCR. Cdh5 mRNA. All LNPs containing 20% ​​EDOPC showed significant activity in all tested tissues. Cdh5 Gene silencing indicates that these new formulations can be used for RNA delivery to endothelial RNA in a variety of tissues. Formulation details are shown in Table 27. The physicochemical properties of LNPs are shown in Table 28. Results are presented in... Figure 20A-20G middle.

[0328] Table 27: Formulations

[0329] Table 28: Physicochemical Properties

[0330] Example 15: 18.5% EDOPC with different PEG lipids To investigate whether the alkyl chain length of PEG-lipids affects the tissue tropism of LNPs containing 18.5% EDOPC, three LNPs with different PEG lipids were prepared and their endothelial delivery efficiency was tested in vivo using a single intravenous administration of 1.0 mg / kg siCdh5.

[0331] Three days after application, tissues were obtained and quantified by RT-qPCR. Cdh5 mRNA. All the LNPs mentioned showed comparable results in all collected tissues. Cdh5 Gene silencing. Formulation details are shown in Table 29. Physicochemical properties of LNPs are shown in Table 30. Results are presented in... Figures 21A-21E middle.

[0332] Table 29: Formulations

[0333] Table 30: Physicochemical Properties

[0334] Example 16: 10% DPTAP with different ionizable lipids To explore the universality of DPTAP for extrahepatic delivery, various ionizable lipids were formulated with 10% DPTAP. Six different ionizable lipid-containing polymeric nippets (LNPs) were prepared, and their endothelial delivery efficiency was tested in vivo using a single intravenous administration of 0.5 mg / kg siCdh5. Three days after administration, tissue samples were obtained, and quantification was performed by RT-qPCR. Cdh5 mRNA. All LNPs containing 10% DPTAP showed significant activity in all tested tissues. Cdh5 Gene silencing indicates that these new formulations can be used for RNA delivery to endothelial RNA in a variety of tissues. Formulation details are shown in Table 31. The physicochemical properties of LNPs are shown in Table 32. Results are presented in... Figures 22A-22F middle.

[0335] Table 31: Formulations

[0336] Table 32: Physicochemical Properties

Claims

1. A composition comprising a plurality of lipid nanoparticles, wherein the lipid nanoparticles comprise an agent, an ionizable lipid, and a fully saturated cationic lipid, wherein the lipid nanoparticles are free of zwitterionic phospholipids.

2. The composition according to claim 1, wherein the lipid nanoparticles have an apparent ionization constant (pKa) of about 4.5 to about 7.

3. The composition according to claim 1 or 2, wherein the lipid nanoparticles have an apparent ionization constant (pKa) of about 5 to about 7.

4. The composition according to any one of claims 1-3, wherein the lipid nanoparticles have an apparent ionization constant (pKa) of about 5.5 to about 7.

5. The composition according to any one of claims 1-4, wherein the lipid nanoparticles have an apparent ionization constant (pKa) of about 6 to about 7.

6. The composition according to any one of claims 1-5, wherein the cationic lipid is less than about 50 mol of the lipid nanoparticles.

7. The composition according to any one of claims 1-6, wherein the cationic lipid is less than about 30 mol of the lipid nanoparticles.

8. The composition according to any one of claims 1-7, wherein the cationic lipid is less than about 20 mol of the lipid nanoparticles.

9. The composition according to any one of claims 1-8, wherein the fully saturated cationic lipid is C 12 -C 18 Fully saturated cationic lipids.

10. The composition according to any one of claims 1-9, further comprising sterols.

11. The composition according to any one of claims 1-10, further comprising PEG-lipids.

12. The composition according to any one of claims 1-11, wherein the pharmaceutical agent is a bioactive agent.

13. The composition according to claim 12, wherein the bioactive agent is a therapeutic agent.

14. The composition of claim 13, wherein the therapeutic agent is a nutritional supplement.

15. The composition according to any one of claims 1-11, wherein the agent is a developer.

16. The composition according to any one of claims 1-11, wherein the pharmaceutical agent is a diagnostic agent.

17. The composition according to any one of claims 1-11, wherein the agent is a nucleic acid.

18. The composition according to claim 17, wherein the nucleic acid is ribonucleic acid.

19. The composition according to claim 17, wherein the nucleic acid is deoxyribonucleic acid.

20. The composition according to claim 17, wherein the nucleic acid is a non-natural nucleic acid.

21. The composition according to any one of claims 1-20, wherein the ionizable lipid is a lipid of formula I or II or a pharmaceutically acceptable salt thereof: (I)； (II), in X is -NR 1 -or -O-; R 1 It is a hydrogen atom, a hydrocarbon group with 6 to 24 atoms, or R 21 -L 1 -R 22 -; R 21 It is a hydrocarbon group having 1 to 24 carbon atoms; L 1 Is it -O(CO)O-, -O(CO)-, -(CO)O-, -O- or ; R 22 It is a divalent hydrocarbon linking group having 1 to 18 carbon atoms; R 2 and R 3 Each is independently a hydrogen atom, a hydrocarbon group having 3 to 24 carbon atoms, or an R... 31 -L 1 -R 32 -; R 31 It is a hydrocarbon group having 1 to 24 carbon atoms; L 2 Is it -O(CO)O-, -O(CO)-, -(CO)O-, -O- or ; R 32 It is a divalent hydrocarbon linking group having 1 to 18 carbon atoms; R 4 R 5 R 6 R 7 R 8 R 9 R 10 R 11 and R 12 Each is independently a hydrogen atom or optionally R 33 Substituted alkyl groups having 1 to 18 carbon atoms, wherein the alkyl group is selected from R 4 and R 5 R 10 and R 5 R 5 and R 12 R 4 and R 6 R 5 and R 6 R 6 and R 7 R 6 and R 10 R 12 and R 7 and R 7 and R 8 One or more pairs of groups can be linked together to form a 4- to 7-membered ring of carbon and nitrogen atoms, said ring may contain oxygen atoms; R 33 It is a hydroxyl group, a carboxyl group, or -NR. 45 R 46 -O(CO)OR 41 -O(CO)-R 42 -(CO)OR 43 -OR 44 Or optionally R 34 Substituted aryl or heteroaryl groups; R 34 It is an alkyl group, hydroxy group, carboxyl group, or -NR group having 1 to 18 carbon atoms. 45 R 46 -O(CO)OR 41 -O(CO)-R 42 -(CO)OR 43 or -OR 44 ; R 41 R 42 R 43 R 44 R 45 and R 46 Each is independently a hydrocarbon group having 1 to 18 carbon atoms; a, b, c, and d are each independent integers from 0 to 3, where a+b is 1 or greater, and c+d is 1 or greater; R 51 and R 52 Each independently is arbitrarily R 35 Substituted alkyl groups having 1 to 18 carbon atoms; R 35 It is a hydroxyl group, -G 20 -CH(R 55 (R) 56 ), -N(R 58 (R) 59 ) or -G 20 -R 60 ; G 20 is-(CO)O-or-O(CO)-; R 55 and R 56 Each is independently hydrogen or an alkyl group having 1 to 18 carbon atoms; R 58 and R 59 Each independently is hydrogen or optionally R 36 Substituted cycloalkyl groups having 3 to 6 carbon atoms; R 60 It is an alkyl group having 1 to 18 carbon atoms or a cycloalkyl group having 3 to 6 carbon atoms; R 36 It is -N(R) 61 (R) 62 ) or -G 20 -R 65 ; R 61 and R 62 Each is independently hydrogen or a cycloalkyl group having 3 to 6 carbon atoms; R 65 It is an alkyl group having 1 to 18 carbon atoms, -L 40 -CH(R 66 (R) 67 ) or -G 20 -R 66 ; L 40 It is a divalent alkyl group containing 1 to 6 carbon atoms; R 66 and R 67 Each is independently an alkyl group containing 1 to 10 carbon atoms or an alkoxy group containing 1 to 10 carbon atoms; L 10 It is a divalent alkyl group containing 1 to 10 carbon atoms; G 30 For -S(CO)N(R 64 )-; R 64 Yes -L 30 -G 20 -CH(R 55 (R) 56 ); a' is 0 or 1; G 10 is G 20 、 -O(CO)O- or -N(R 63 )C(O)-; R 63 It is an alkyl group containing 1 to 18 carbon atoms; L 20 It is a divalent alkyl group containing 1 to 6 carbon atoms; b' is 0 or 1; R 53 R 54 and R 57 Each independently is hydrogen or optionally R 36 The substituted alkyl group containing 1 to 18 carbon atoms; and L 30 It is a single bond or an alkyl group containing 1 to 18 carbon atoms.

22. The composition according to any one of claims 1-20, wherein the ionizable lipid is a lipid of formula VII or a pharmaceutically acceptable salt thereof: (VII), in R 1 and R 2 Each is independently a hydrocarbon group having 1 to 18 carbon atoms, and R 3 It is a hydrocarbon group having 2 to 8 carbon atoms, wherein R 1 R 2 and R 3 The hydrocarbon groups represented are each independently and optionally surrounded by one or more groups selected from -OH, COOH, -NR. 51 R 52 -OC(O)OR 53 -C(O)OR 54 -OC(O)-R 55 and -OR 56 Substituents of the substituents; R 4 It is a hydrocarbon group having 1 to 8 carbon atoms; R 5 and R 6 Each is independently a hydrocarbon group having 1 to 8 carbon atoms, or -R 8 -L 1 -R 9 Exclude R 5 and R 6 Both cases involve hydrocarbon groups having 1 to 8 carbon atoms; R 7 Yes -R 10 -L 2 -R 11 -L 3 -R 12 ; R 51 and R 52 Each is an independent hydrocarbon group having 1 to 8 carbon atoms; R 53 R 54 R 55 and R 56 Each is an independent hydrocarbon group having 1 to 24 carbon atoms; By R 53 R 54 R 55 and R 56 Each of the hydrocarbon groups represented is independently and optionally surrounded by an aryl group having 6 to 20 carbon atoms or -SR 58 The aryl group having 6 to 20 carbon atoms is optionally replaced by -OH, COOH, or -NR. 51 R 52 -OC(O)OR 53 -C(O)OR 54 -OC(O)-R 55 -OR 56 or -R 59 -R 57 Replace; among them R 58 and R 59 Each is an independent hydrocarbon group having 1 to 12 carbon atoms; R 57 -OH, COOH, -NR 61 R 62 -OC(O)OR 63 -C(O)OR 64 -OC(O)-R 65 or -OR 66 ; R 61 and R 62 Each is an independent hydrocarbon group having 1 to 8 carbon atoms; R 63 R 64 R 65 and R 66 Each is an independent hydrocarbon group having 1 to 24 carbon atoms; By R 63 R 64 R 65 and R 66 Each of the hydrocarbon groups represented is independently and optionally surrounded by an aryl group having 6 to 20 carbon atoms or -SR 68 The aryl group having 6 to 20 carbon atoms can be replaced by -OH, COOH, -NR, etc. 61 R 62 -OC(O)OR 63 -C(O)OR 64 -OC(O)-R 65 -OR 66 or -R 69 -R 67 replace; R 68 and R 69 Each is an independent hydrocarbon group having 1 to 12 carbon atoms; R 67 -OH, COOH, -NR 61 R 62 -OC(O)OR 63 -C(O)OR 64 -OC(O)-R 65 or -OR 66 ; L 1 L 2 and L 3 Each can be independently represented as -OC(O)O-, -C(O)O-, -OC(O)-, or -O-; R 8 It is a hydrocarbon group having 1 to 12 carbon atoms; R 9 It is a hydrocarbon group having 1 to 24 carbon atoms; R 10 It is a hydrocarbon group having 1 to 8 carbon atoms; R 11 It is a hydrocarbon group having 1 to 24 carbon atoms; R 12 It is a hydrocarbon group having 1 to 24 carbon atoms; By R 9 and R 12 The hydrocarbon groups represented are each independently and optionally replaced by aryl groups, -OC(O)OR 53 -C(O)OR 54 -OC(O)-R 55 or -SR 58 Replace; and By R 11 The hydrocarbon groups represented are each independently and optionally labeled -OC(O)OR 53 -C(O)OR 54 or -OC(O)-R 55 replace.

23. The composition according to any one of claims 1-20, wherein the ionizable lipid is a lipid of formula VIII or a pharmaceutically acceptable salt thereof: (VIII), in R 1 and R 2 Each is independently a hydrocarbon group having 1 to 18 carbon atoms, and R 3 It is a hydrocarbon group having 2 to 8 carbon atoms, wherein R 1 R 2 and R 3 The hydrocarbon groups represented are each independently and optionally converted by -OH, COOH, or -NR. 51 R 52 -OC(O)OR 53 -C(O)OR 54 -OC(O)-R 55 or -OR 56 replace; R 4 It is a hydrocarbon group having 1 to 8 carbon atoms; R 5 and R 6 Each is independently a hydrocarbon group having 1 to 8 carbon atoms, or -R 8 -L 1 -R 9 Exclude R 5 and R 6 Both cases involve hydrocarbon groups having 1 to 8 carbon atoms; L 1 and L 4 Each can be independently represented as -OC(O)O-, -C(O)O-, -OC(O)-, or -O-; R 8 It is a hydrocarbon group having 1 to 12 carbon atoms; R 9 It is a hydrocarbon group having 1 to 24 carbon atoms, wherein R 9 The hydrocarbon group is optionally replaced by an aryl group, -OC(O)OR 53 -C(O)OR 54 -OC(O)-R 55 or -SR 58 replace; R 51 and R 52 Each is an independent hydrocarbon group having 1 to 8 carbon atoms; R 53 R 54 R 55 and R 56 Each is an independent hydrocarbon group having 1 to 24 carbon atoms; By R 53 R 54 R 55 and R 56 Each of the hydrocarbon groups represented is independently and optionally surrounded by an aryl group having 6 to 20 carbon atoms or -SR 58 The aryl group having 6 to 20 carbon atoms is optionally replaced by -OH, COOH, or -NR. 51 R 52 -OC(O)OR 53 -C(O)OR 54 -OC(O)-R 55 -OR 56 or -R 59 -R 57 replace; R 58 and R 59 Each is an independent hydrocarbon group having 1 to 12 carbon atoms; R 57 -OH, COOH, -NR 51 R 52 -OC(O)OR 53 -C(O)OR 54 -OC(O)-R 55 or -OR 56 ; R 13 It is a hydrocarbon group having 1 to 8 carbon atoms; R 14 Yes -R 15 -L 5 -R 16 , where R 15 It is a hydrocarbon group with 1 to 24 carbon atoms, L 5 It is -OC(O)O-, -C(O)O-, -OC(O)-, or -O-, and R 16 It is a hydrocarbon group having 1 to 24 carbon atoms; By R 15 The hydrocarbon group representing 1 to 24 carbon atoms is optionally represented by -OC(O)OR 53 -C(O)OR 54 or -OC(O)-R 55 Replace; and By R 16 The hydrocarbon group having 1 to 24 carbon atoms is optionally replaced by an aryl group having 6 to 20 carbon atoms, -OC(O)OR 53 -C(O)OR 54 -OC(O)-R 55 or -SR 58 replace.

24. The composition according to any one of claims 1-20, wherein the ionizable lipid is a lipid of formula IX or a pharmaceutically acceptable salt thereof: (IX), in R 1 and R 2 Each is independently a hydrocarbon group having 1 to 18 carbon atoms, and R 3 It is a hydrocarbon group having 2 to 8 carbon atoms, wherein R 1 R 2 and R 3 The hydrocarbon groups represented are optionally replaced by -OH, COOH, or -NR. 51 R 52 -OC(O)OR 53 -C(O)OR 54 -OC(O)-R 55 or -OR 56 replace; R 4 and R 8 Each is independently a hydrocarbon having 1 to 8 carbon atoms; R 21 and R 22 Each is independently a hydrocarbon group having 1 to 18 carbon atoms; R 23 and R 24 Each is an independent hydrocarbon group having 1 to 12 carbon atoms; R 25 and R 26 Each is an independent hydrocarbon group having 1 to 24 carbon atoms; L 21 and L 22 Each can be independently represented as -OC(O)O-, -C(O)O-, -OC(O)-, or -O-; By R 25 and R 26 The hydrocarbon groups represented are each independently and optionally replaced by an aryl group having 6 to 20 carbon atoms, -OC(O)OR 53 -C(O)OR 54 -OC(O)-R 55 or -SR 58 Substitution, wherein the aryl group having 6 to 20 carbon atoms is optionally replaced by OH, COOH, -NR 51 R 52 -OC(O)OR 53 -C(O)OR 54 -OC(O)-R 55 -OR 56 or -R 59 -R 57 Replace, and R 51 and R 52 Each is an independent hydrocarbon group having 1 to 8 carbon atoms; R 53 R 54 R 55 and R 56 Each is independently a hydrocarbon group having 1 to 18 carbon atoms; - R 57 -OH, COOH, -NR 51 R 52 -OC(O)OR 53 -C(O)OR 54 -OC(O)-R 55 or -OR 56 ;and R 58 and R 59 Each is an independent hydrocarbon group having 1 to 12 carbon atoms.

25. The composition according to any one of claims 1-20, wherein the ionizable lipid is a lipid of formula X or a pharmaceutically acceptable salt thereof: (X)， in R 1 and R 2 Each is independently a hydrocarbon group having 1 to 18 carbon atoms, and R 3 It is a hydrocarbon group having 2 to 8 carbon atoms, wherein R 1 R 2 and R 3 The hydrocarbon groups represented are each independently and optionally converted by -OH, COOH, or -NR. 51 R 52 -OC(O)OR 53 -C(O)OR 54 -OC(O)-R 55 or -OR 56 replace; R 4 and R 8 Each is an independent hydrocarbon group having 1 to 8 carbon atoms; R 31 R 32 R 33 and R 34 Each is independently a hydrocarbon group having 1 to 12 carbon atoms. R 35 R 36 R 37 and R 38 Each is an independent hydrocarbon group having 1 to 24 carbon atoms; L 31 L 32 L 33 and L 34 Each can be independently represented as -OC(O)O-, -C(O)O-, -OC(O)-, or -O-; By R 35 R 36 R 37 and R 38 The hydrocarbon groups represented are each independently and optionally replaced by an aryl group having 6 to 20 carbon atoms, -OC(O)OR 53 -C(O)OR 54 -OC(O)-R 55 or SR 58 Substitution, wherein the aryl group having 6 to 20 carbon atoms is optionally replaced by OH, COOH, -NR 51 R 52 -OC(O)OR 53 -C(O)OR 54 -OC(O)-R 55 -OR 56 or -R 59 -R 57 Replace, and R 51 and R 52 Each is an independent hydrocarbon group having 1 to 8 carbon atoms; R 53 R 54 R 55 and R 56 Each is independently a hydrocarbon group having 1 to 18 carbon atoms; R 57 -OH, COOH, -NR 51 R 52 -OC(O)OR 53 -C(O)OR 54 -OC(O)-R 55 or -OR 56 ;and R 58 and R 59 Each is an independent hydrocarbon group having 1 to 12 carbon atoms.

26. The composition according to any one of claims 1-21, wherein the ionizable lipid is a lipid of formula III or a pharmaceutically acceptable salt thereof: (III), in R 2 and R 3 Each is independently a hydrogen atom, a hydrocarbon group having 3 to 24 carbon atoms, or an R... 31 -L 2 -R 32 -; R 31 It is a hydrocarbon group having 1 to 24 carbon atoms; L 2 Is it -O(CO)O-, -O(CO)-, -(CO)O-, -O- or ; R 32 It is a divalent hydrocarbon linking group having 1 to 18 carbon atoms; R 5 It is a hydrogen atom or optionally R 33 Substituted alkyl groups having 1 to 18 carbon atoms; R 7 and R 8 Each is independently a hydrogen atom or optionally R 33 Substituted alkyl groups having 1 to 18 carbon atoms; R 33 It is a hydroxyl group, a carboxyl group, or -NR. 45 R 46 -O(CO)OR 41 -O(CO)-R 42 -(CO)OR 43 -OR 44 Or optionally R 34 Substituted aryl or heteroaryl groups; R 34 It is an alkyl group, hydroxy group, carboxyl group, or -NR group having 1 to 18 carbon atoms. 45 R 46 -O(CO)OR 41 -O(CO)-R 42 -(CO)OR 43 or -OR 44 ; R 41 R 42 R 43 R 44 R 45 and R 46 Each is independently a hydrocarbon group having 1 to 18 carbon atoms; and e is an integer selected from 2 or 3.

27. The composition according to any one of claims 1-21 and 26, wherein the ionizable lipid is selected from: and 。 28. The composition according to any one of claims 1-20 and 22, wherein the ionizable lipid is selected from: and 。 29. The composition according to any one of claims 1-28, wherein the cationic lipid is a lipid according to formula IV: (IV), in: R 101 and R 102 Each is independently an optional substitution (C8-C) 24 )alkyl or optionally substituted (C8-C 24 alkenyl; R 103 In each case, it is independently an optionally substituted (C1-C6) alkyl group; R 104 It is an optionally substituted (C1-C6) alkyl group; and X⁻ is a monovalent anion.

30. The composition according to any one of claims 1-29, wherein the cationic lipid is 1,2-dilauryl- sn -Glyceryl-O-ethyl-3-phosphocholine (EDLPC), 1,2-dimyristoyl- sn -Glyceryl-O-ethyl-3-phosphocholine (EDMPC), 1,2-dimyristoleoyl- sn -Glyceryl-O-ethyl-3-phosphocholine (14:1 EPC), 1,2-dipalmitoyl- sn -Glyceryl-O-ethyl-3-phosphocholine (EDPPC), 1,2-distearate- sn -Glyceryl-O-ethyl-3-phosphocholine (EDSPC), 1,2-dioleoyl- sn -Glyceryl-3-ethylphosphorylcholine (EDOPC) or 1-palmitoyl-2-oleoyl- sn -Glyceryl-O-ethyl-3-phosphocholine (16:0-18:1 EPC).

31. The composition according to any one of claims 1-30, wherein the cationic lipid is EDPPC.

32. The composition according to any one of claims 1-28, wherein the cationic lipid is a lipid according to formula V: (V)， in: R 101 and R 102 Each is independently an optional substitution (C8-C) 24 )alkyl or optionally substituted (C8-C 24 alkenyl; R 103 In each case, it is independently an optionally substituted (C1-C6) alkyl group; and X⁻ is a monovalent anion.

33. The composition according to any one of claims 1-28 and 32, wherein the cationic lipid is selected from 1,2-dimyristoyl-3-trimethylammonium-propane (DMTAP), 1,2-dipalmitoyl-3-trimethylammonium-propane (DPTAP), 1,2-distearate-3-trimethylammonium-propane (DSTAP), 1,2-dioleoyl-3-trimethylammonium-propane (DOTAP) and N-(2-hydroxyethyl)-N,N-dimethyl-2,3-bis(oleoyloxy)prop-1-ammonium (DORI).

34. The composition according to any one of claims 1-28, 32 and 33, wherein the cationic lipid is DPTAP.

35. The composition according to any one of claims 1-28, wherein the cationic lipid is a lipid according to formula VI: (WE), in: R 101 and R 102 Each is independently an optional substitution (C8-C) 24 )alkyl or optionally substituted (C8-C 24 alkenyl; R 105 Each is independently a (C1-C6) alkyl group; and X⁻ is a monovalent anion.

36. The composition according to any one of claims 1-28 and 35, wherein the cationic lipid is 。 37. A composition comprising a plurality of lipid nanoparticles, wherein the lipid nanoparticles comprise a pharmaceutical agent, an ionizable lipid, and a cationic lipid, wherein the cationic lipid is a lipid according to formula VI: (WE), in: R 101 and R 102 Each is independently an optional substitution (C8-C) 24 )alkyl or optionally substituted (C8-C 24 alkenyl; R 105 Each is independently a (C1-C6) alkyl group; and X⁻ is a monovalent anion.

38. The composition of claim 37, wherein the cationic lipid is 。 39. A composition comprising a plurality of lipid nanoparticles, wherein the lipid nanoparticles comprise a pharmaceutical agent, an ionizable lipid, and a cationic lipid, wherein the ionizable lipid is a lipid of formula I or a pharmaceutically acceptable salt thereof: (I)， in X is -NR 1 -or -O-; R 1 It is a hydrogen atom, a hydrocarbon group with 6 to 24 atoms, or R 21 -L 1 -R 22 -; R 21 It is a hydrocarbon group having 1 to 24 carbon atoms; L 1 Is it -O(CO)O-, -O(CO)-, -(CO)O-, -O- or ; R 22 It is a divalent hydrocarbon linking group having 1 to 18 carbon atoms; R 2 and R 3 Each is independently a hydrogen atom, a hydrocarbon group having 3 to 24 carbon atoms, or an R... 31 -L 1 -R 32 -; R 31 It is a hydrocarbon group having 1 to 24 carbon atoms; L 2 Is it -O(CO)O-, -O(CO)-, -(CO)O-, -O- or ; R 32 It is a divalent hydrocarbon linking group having 1 to 18 carbon atoms; R 4 R 5 R 6 R 7 R 8 R 9 R 10 R 11 and R 12 Each is independently a hydrogen atom or optionally R 33 Substituted alkyl groups having 1 to 18 carbon atoms, wherein the alkyl group is selected from R 4 and R 5 R 10 and R 5 R 5 and R 12 R 4 and R 6 R 5 and R 6 R 6 and R 7 R 6 and R 10 R 12 and R 7 and R 7 and R 8 One or more pairs of groups can be linked together to form a 4- to 7-membered ring of carbon and nitrogen atoms, said ring may contain oxygen atoms; R 33 It is a hydroxyl group, a carboxyl group, or -NR. 45 R 46 -O(CO)OR 41 -O(CO)-R 42 -(CO)OR 43 -OR 44 Or optionally R 34 Substituted aryl or heteroaryl groups; R 34 It is an alkyl group, hydroxy group, carboxyl group, or -NR group having 1 to 18 carbon atoms. 45 R 46 -O(CO)OR 41 -O(CO)-R 42 -(CO)OR 43 or -OR 44 ; R 41 R 42 R 43 R 44 R 45 and R 46 Each is independently a hydrocarbon group having 1 to 18 carbon atoms; and a, b, c, and d are each independent integers from 0 to 3, where a+b is 1 or greater, and c+d is 1 or greater.

40. The composition of claim 39, wherein the ionizable lipid is a lipid of formula III or a pharmaceutically acceptable salt thereof: (III), in R 2 and R 3 Each is independently a hydrogen atom, a hydrocarbon group having 3 to 24 carbon atoms, or an R... 31 -L 2 -R 32 -; R 31 It is a hydrocarbon group having 1 to 24 carbon atoms; L 2 Is it -O(CO)O-, -O(CO)-, -(CO)O-, -O- or ; R 32 It is a divalent hydrocarbon linking group having 1 to 18 carbon atoms; R 5 It is a hydrogen atom or optionally R 33 Substituted alkyl groups having 1 to 18 carbon atoms; R 33 It is a hydroxyl group, a carboxyl group, or -NR. 45 R 46 -O(CO)OR 41 -O(CO)-R 42 -(CO)OR 43 -OR 44 Or optionally R 34 Substituted aryl or heteroaryl groups; R 7 and R 8 Each is independently a hydrogen atom or optionally R 33 Substituted alkyl groups having 1 to 18 carbon atoms; R 34 It is an alkyl group, hydroxy group, carboxyl group, or -NR group having 1 to 18 carbon atoms. 45 R 46 -O(CO)OR 41 -O(CO)-R 42 -(CO)OR 43 or -OR 44 ; R 41 R 42 R 43 R 44 R 45 and R 46 Each is independently a hydrocarbon group having 1 to 18 carbon atoms; and e is an integer selected from 2 or 3.

41. The composition according to claim 39 or 40, wherein the ionizable lipid is selected from: and 。 42. A composition comprising a plurality of lipid nanoparticles, wherein the lipid nanoparticles comprise a pharmaceutical agent, an ionizable lipid, and a cationic lipid, wherein the ionizable lipid is a lipid according to Formula VII or a pharmaceutically acceptable salt thereof: (VII), in R 1 and R 2 Each is independently a hydrocarbon group having 1 to 18 carbon atoms, and R 3 It is a hydrocarbon group having 2 to 8 carbon atoms, wherein R 1 R 2 and R 3 The hydrocarbon groups represented are each independently and optionally surrounded by one or more groups selected from -OH, COOH, -NR. 51 R 52 -OC(O)OR 53 -C(O)OR 54 -OC(O)-R 55 and -OR 56 Substituents of the substituents; R 4 It is a hydrocarbon group having 1 to 8 carbon atoms; R 5 and R 6 Each is independently a hydrocarbon group having 1 to 8 carbon atoms, or -R 8 -L 1 -R 9 Exclude R 5 and R 6 Both cases involve hydrocarbon groups having 1 to 8 carbon atoms; R 7 Yes -R 10 -L 2 -R 11 -L 3 -R 12 ; R 51 and R 52 Each is an independent hydrocarbon group having 1 to 8 carbon atoms; R 53 R 54 R 55 and R 56 Each is an independent hydrocarbon group having 1 to 24 carbon atoms; By R 53 R 54 R 55 and R 56 Each of the hydrocarbon groups represented is independently and optionally surrounded by an aryl group having 6 to 20 carbon atoms or -SR 58 The aryl group having 6 to 20 carbon atoms is optionally replaced by -OH, COOH, or -NR. 51 R 52 -OC(O)OR 53 -C(O)OR 54 -OC(O)-R 55 -OR 56 or -R 59 -R 57 Replace; among them R 58 and R 59 Each is an independent hydrocarbon group having 1 to 12 carbon atoms; R 57 -OH, COOH, -NR 61 R 62 -OC(O)OR 63 -C(O)OR 64 -OC(O)-R 65 or -OR 66 ; R 61 and R 62 Each is an independent hydrocarbon group having 1 to 8 carbon atoms; R 63 R 64 R 65 and R 66 Each is an independent hydrocarbon group having 1 to 24 carbon atoms; By R 63 R 64 R 65 and R 66 Each of the hydrocarbon groups represented is independently and optionally surrounded by an aryl group having 6 to 20 carbon atoms or -SR 68 The aryl group having 6 to 20 carbon atoms can be replaced by -OH, COOH, -NR, etc. 61 R 62 -OC(O)OR 63 -C(O)OR 64 -OC(O)-R 65 -OR 66 or -R 69 -R 67 replace; R 68 and R 69 Each is an independent hydrocarbon group having 1 to 12 carbon atoms; R 67 -OH, COOH, -NR 61 R 62 -OC(O)OR 63 -C(O)OR 64 -OC(O)-R 65 or -OR 66 ; L 1 L 2 and L 3 Each can be independently represented as -OC(O)O-, -C(O)O-, -OC(O)-, or -O-; R 8 It is a hydrocarbon group having 1 to 12 carbon atoms; R 9 It is a hydrocarbon group having 1 to 24 carbon atoms; R 10 It is a hydrocarbon group having 1 to 8 carbon atoms; R 11 It is a hydrocarbon group having 1 to 24 carbon atoms; R 12 It is a hydrocarbon group having 1 to 24 carbon atoms; By R 9 and R 12 The hydrocarbon groups represented are each independently and optionally replaced by aryl groups, -OC(O)OR 53 -C(O)OR 54 -OC(O)-R 55 or -SR 58 Replace; and By R 11 The hydrocarbon groups represented are each independently and optionally labeled -OC(O)OR 53 -C(O)OR 54 or -OC(O)-R 55 replace.

43. The composition of claim 42, wherein the ionizable lipid is selected from: and 。 44. A composition comprising a plurality of lipid nanoparticles, wherein the lipid nanoparticles comprise a pharmaceutical agent, an ionizable lipid, and at least one of EDPPC and DPTAP, wherein the ionizable lipid is a lipid of formula I or II or a pharmaceutically acceptable salt thereof: (I)； (II), in X is -NR 1 -or -O-; R 1 It is a hydrogen atom, a hydrocarbon group with 6 to 24 atoms, or R 21 -L 1 -R 22 -; R 21 It is a hydrocarbon group having 1 to 24 carbon atoms; L 1 Is it -O(CO)O-, -O(CO)-, -(CO)O-, -O- or ; R 22 It is a divalent hydrocarbon linking group having 1 to 18 carbon atoms; R 2 and R 3 Each is independently a hydrogen atom, a hydrocarbon group having 3 to 24 carbon atoms, or an R... 31 -L 1 -R 32 -; R 31 It is a hydrocarbon group having 1 to 24 carbon atoms; L 2 Is it -O(CO)O-, -O(CO)-, -(CO)O-, -O- or ; R 32 It is a divalent hydrocarbon linking group having 1 to 18 carbon atoms; R 4 R 5 R 6 R 7 R 8 R 9 R 10 R 11 and R 12 Each is independently a hydrogen atom or optionally R 33 Substituted alkyl groups having 1 to 18 carbon atoms, wherein the alkyl group is selected from R 4 and R 5 R 10 and R 5 R 5 and R 12 R 4 and R 6 R 5 and R 6 R 6 and R 7 R 6 and R 10 R 12 and R 7 and R 7 and R 8 One or more pairs of groups can be linked together to form a 4- to 7-membered ring of carbon and nitrogen atoms, said ring may contain oxygen atoms; R 33 It is a hydroxyl group, a carboxyl group, or -NR. 45 R 46 -O(CO)OR 41 -O(CO)-R 42 -(CO)OR 43 -OR 44 Or optionally R 34 Substituted aryl or heteroaryl groups; R 34 It is an alkyl group, hydroxy group, carboxyl group, or -NR group having 1 to 18 carbon atoms. 45 R 46 -O(CO)OR 41 -O(CO)-R 42 -(CO)OR 43 or -OR 44 ; R 41 R 42 R 43 R 44 R 45 and R 46 Each is independently a hydrocarbon group having 1 to 18 carbon atoms; a, b, c, and d are each independent integers from 0 to 3, where a+b is 1 or greater, and c+d is 1 or greater; R 51 and R 52 Each independently is arbitrarily R 35 Substituted alkyl groups having 1 to 18 carbon atoms; R 35 It is a hydroxyl group, -G 20 -CH(R 55 (R) 56 ), -N(R 58 (R) 59 ) or -G 20 -R 60 ; G 20 is-(CO)O-or-O(CO)-; R 55 and R 56 Each is independently hydrogen or an alkyl group having 1 to 18 carbon atoms; R 58 and R 59 Each independently is hydrogen or optionally R 36 Substituted cycloalkyl groups having 3 to 6 carbon atoms; R 60 It is an alkyl group having 1 to 18 carbon atoms or a cycloalkyl group having 3 to 6 carbon atoms; R 36 It is -N(R) 61 (R) 62 ) or -G 20 -R 65 ; R 61 and R 62 Each is independently hydrogen or a cycloalkyl group having 3 to 6 carbon atoms; R 65 It is an alkyl group having 1 to 18 carbon atoms, -L 40 -CH(R 66 (R) 67 ) or -G 20 -R 66 ; L 40 It is a divalent alkyl group containing 1 to 6 carbon atoms; R 66 and R 67 Each is independently an alkyl group containing 1 to 10 carbon atoms or an alkoxy group containing 1 to 10 carbon atoms; L 10 It is a divalent alkyl group containing 1 to 10 carbon atoms; G 30 For -S(CO)N(R 64 )-; R 64 Yes -L 30 -G 20 -CH(R 55 (R) 56 ); a' is 0 or 1; G 10 is G 20 、 -O(CO)O- or -N(R 63 )C(O)-; R 63 It is an alkyl group containing 1 to 18 carbon atoms; L 20 It is a divalent alkyl group containing 1 to 6 carbon atoms; b' is 0 or 1; R 53 R 54 and R 57 Each independently is hydrogen or optionally R 36 The substituted alkyl group containing 1 to 18 carbon atoms; and L 30 It is a single bond or an alkyl group containing 1 to 18 carbon atoms.

45. The composition according to any one of claims 1-44, wherein the pharmaceutical agent is a bioactive agent.

46. ​​The composition according to claim 45, wherein the bioactive agent is a therapeutic agent.

47. The composition of claim 46, wherein the therapeutic agent is a nutritional supplement.

48. The composition according to any one of claims 1-44, wherein the agent is a developer.

49. The composition according to any one of claims 1-44, wherein the pharmaceutical agent is a diagnostic agent.

50. The composition according to any one of claims 1-44, wherein the agent is a nucleic acid.

51. The composition according to claim 50, wherein the nucleic acid is ribonucleic acid.

52. The composition according to claim 50, wherein the nucleic acid is deoxyribonucleic acid.

53. The composition according to claim 50, wherein the nucleic acid is a non-natural nucleic acid.

54. A method of delivering a drug to one or more of the brain, heart, muscles, and kidneys of an individual in need, the method comprising systemically administering to the individual a composition comprising a plurality of lipid nanoparticles, wherein the lipid nanoparticles comprise the drug, ionizable lipids, and fully saturated cationic lipids.

55. The method of claim 54, wherein the agent is delivered to the brain.

56. The method of claim 54, wherein the agent is delivered to the heart.

57. The method of claim 54, wherein the agent is delivered to muscle.

58. The method of claim 54, wherein the agent is delivered to the kidney.

59. The method according to any one of claims 54-58, wherein the ionizable lipid is a lipid of formula (I).

60. The method according to any one of claims 54-59, wherein the ionizable lipid is a lipid of formula (III).

61. The method according to any one of claims 54-60, wherein the ionizable lipid is a lipid of formula (II).

62. The method according to any one of claims 54-58, wherein the ionizable lipid is a lipid of formula (VII).

63. The method according to any one of claims 54-58 and 62, wherein the ionizable lipid is a lipid of formula (VIII).

64. The method according to any one of claims 54-58 and 62, wherein the ionizable lipid is a lipid of formula (IX).

65. The method according to any one of claims 54-58 and 62, wherein the ionizable lipid is a lipid of formula (X).

66. The method according to any one of claims 54-65, wherein the cationic lipid is a lipid of formula (IV).

67. The method according to any one of claims 54-65, wherein the cationic lipid is a lipid of formula (V).

68. The method according to any one of claims 54-65, wherein the cationic lipid is a lipid of formula (VI).

69. A method of delivering a pharmaceutical agent to an individual in need, the method comprising administering to the individual the composition of any one of claims 1-53.

70. A method for treating an individual suffering from a disease, disorder, or ailment that is beneficially treatable by a pharmaceutical agent, the method comprising administering to the individual a therapeutically effective amount of the composition of any one of claims 1-53.

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