Lipid nanoparticles targeting the splen

Abstract

The invention relates to lipid nanoparticles containing a steroid compound, as well as to the production and use of such lipid nanoparticles. Such lipid nanoparticles are suitable for the in vivo delivery of payloads, such as nucleic acids, to non-liver organs (e.g., the spleen) for the treatment or prevention of certain diseases or disorders, particularly diseases of the spleen.

Description

Spleen-targeted lipid nanoparticles 1. Technical Field

[0001] The present application relates to the field of drug therapy, in particular to lipid nanoparticles. The application also relates to the preparation of the lipid nanoparticles, and the use of the lipid nanoparticles for targeted delivery of bioactive molecules, such as nucleic acids (e.g., mRNA, miRNA, siRNA, saRNA, ASO, DNA) and polypeptides (e.g., antibodies) to certain organs, in particular the spleen. 2. Background Technology

[0002] In the past few years, nucleic acid-based gene therapy has made rapid progress in various fields such as the treatment of infectious diseases and cancer. Lipid nanoparticles (LNPs) are one of the most advanced non-viral delivery platforms that can safely and effectively deliver nucleic acid drugs (such as mRNA or siRNA) to specific target organs of interest while also protecting them from degradation such as that caused by nucleases. LNPs have several advantages, including high encapsulation efficiency, good cell transfection efficiency, strong tissue penetration, and relatively low cytotoxicity and immunogenicity. Currently, LNPs have been successfully used in several commercial products. For example, FDA-approved drugs such as the mRNA COVID-19 vaccine developed by Moderna and Pfizer-BioNTech, and the siRNA drug Onpattro developed by Alnylam, all utilize the LNP drug delivery platform.

[0003] The study found that after entering the body's circulation, the PEGylated lipids dissociate from the LNP nanoparticles to allow ApoE to adsorb onto the LNP surface, forming an effective ligand for liver targeting. The LNP then binds to lipoprotein receptors on the surface of hepatocytes, triggering endocytosis. Therefore, current LNP delivery systems mainly exhibit passive liver targeting. In order to fully unleash the therapeutic potential of nucleic acid therapy, expand its application to different types of diseases, and minimize toxic side effects on healthy organs, it is necessary to develop LNP delivery systems that target non-hepatic organs.

[0004] The present application provides certain steroid compounds that provide targeted delivery of payloads to non-hepatic organs, particularly the spleen.

[0005] 3. Summary of the Invention

[0006] The present invention provides a spleen-targeted lipid nanoparticle, which has a better delivery effect to the spleen than to other organs such as the liver or lung.

[0007] In one embodiment, the lipid nanoparticles contain a steroid compound and an ionizable lipid.

[0008] In one embodiment, the molar percentage of the steroid compound to the total lipids is 3-15 mol%, preferably 5-10 mol%, for example 3 mol%, 4 mol%, 5 mol%, 6 mol%, 7 mol%, 8 mol%, 9 mol%, 10 mol%, 11 mol%, 12 mol%, 13 mol%, 14 mol% or 15 mol%.

[0009] In one embodiment, the molar percentage of the ionizable lipid is 35-70 mol%, more preferably 40-60 mol%, such as 35 mol%, 40 mol%, 45 mol%, 50 mol%, 55 mol% or 60 mol%.

[0010] In one embodiment, the spleen targeting lipid nanoparticles further comprise a phospholipid.

[0011] In a preferred embodiment, the phospholipid is DSPC, DMPC, DOPC, DPPC, POPC, DOPE, DMPE, POPOE or DPPE.

[0012] In a preferred embodiment, the molar percentage of the phospholipid is 5-30 mol%, preferably 5-20 mol%, more preferably 5-15 mol%, for example 5 mol%, 6 mol%, 7 mol%, 8 mol%, 9 mol%, 10 mol%, 11 mol%, 12 mol%, 13 mol%, 14 mol% or 15 mol%.

[0013] In one embodiment, the spleen-targeted lipid nanoparticles further comprise structural lipids, such as cholesterols.

[0014] In a preferred embodiment, the structured lipid is selected from the group consisting of cholesterol, campesterol, stigmasterol, sitosterol, brassicasterol, ergosterol, solanine, ursolic acid, α-tocopherol, β-sitosterol, avenasterol and rapeseedsterol.

[0015] In a preferred embodiment, the molar percentage of the structured lipid is 10-80 mol%, more preferably 15-70 mol%, more preferably 30-50 mol%, more preferably 30-40 mol%, for example 30 mol%, 35 mol%, 40 mol%, 45 mol%, 50 mol%, 55 mol% or 60 mol%.

[0016] In one embodiment, the spleen-targeted lipid nanoparticles further comprise a polymer-conjugated lipid, such as a pegylated lipid.

[0017] In a preferred embodiment, the PEGylated lipid comprises a PEGylated moiety having a molecular weight of about 1000 Da to about 20,000 Da, about 1000 Da to about 5000 Da, or about 1000 Da to about 2000 Da.

[0018] In a preferred embodiment, the polymer-conjugated lipid is ALC-0159, DMG-PEG2000, DMPE-PEG1000, DPPE-PEG1000, DSPE-PEG1000, DOPE-PEG1000, ceramide-PEG2000, DMPE-PEG2000, DPPE-PEG2000, DSPE-PEG2000, DSPE-PEG2000-mannose, ceramide-PEG5000, DSPE-PEG5000 or DSPE-PEG2000 amine.

[0019] In a preferred embodiment, the molar percentage of the polymer-conjugated lipid is 0.25-10 mol%, more preferably 0.5-5 mol%, more preferably 0.5-3 mol%, for example 0.5 mol%, 0.6 mol%, 0.7 mol%, 0.8 mol%, 0.9 mol%, 1.0 mol%, 1.1 mol%, 1.2 mol%, 1.3 mol%, 1.4 mol%, 1.5 mol%, 1.6 mol%, 1.7 mol%, 1.8 mol%. l%, 1.9mol%, 2.0mol%, 2.1mol%, 2.2mol%, 2.3mol%, 2.4mol%, 2.5mol%, 2.6mol%, 2.7mol%, 2.8mol%, 2.9mol %, 3.0 mol%, 3.1 mol%, 3.2 mol%, 3.3 mol%, 3.4 mol%, 3.5 mol%, 3.6 mol%, 3.7 mol%, 3.8 mol%, 3.9 mol% or 4.0 mol%.

[0020] In one embodiment, the average particle size of the spleen-targeted lipid nanoparticles is 50-800 nm, more preferably 50-400 nm, more preferably 80-200 nm, for example 80, 90, 100, 110, 120, 130, 140, 150, 160, 170, 180, 190 or 200 nm.

[0021] In one embodiment, the lipid nanoparticles contain: (1) a steroid compound; (2) an ionizable lipid; (3) a phospholipid compound; (4) a structural lipid; and (5) a polymer-conjugated lipid.

[0022] In a more preferred embodiment, the lipid nanoparticles further contain a payload.

[0023] In one embodiment, the payload is a therapeutic, prophylactic, or diagnostic agent.

[0024] In one embodiment, the payload is a nucleic acid.

[0025] In a preferred embodiment, the nucleic acid is an antisense oligonucleotide (ASO), DNA or RNA, optionally wherein the RNA is RNA interference (RNAi), small interfering RNA (siRNA), short hairpin RNA (shRNA), antisense RNA (aRNA), messenger RNA (mRNA), modified messenger RNA (mmRNA), long non-coding RNA (lncRNA), microRNA (miRNA), small activating RNA (saRNA), poly-coding nucleic acid (MCNA), polymer-coded nucleic acid (PCNA), guide RNA (gRNA), CRISPR RNA (crRNA) or any other RNA in a ribozyme.

[0026] In a preferred embodiment, the spleen-targeted lipid nanoparticles include the following components in molar percentages:

[0027] 1) steroid compound 3-15 mol%;

[0028] 2) ionizable lipids 35-70 mol%;

[0029] 3) 5-30 mol% phospholipids;

[0030] 4) structural lipids 10-80 mol%; and

[0031] 5) Polymer-conjugated lipid 0.25-10 mol%.

[0032] In a preferred embodiment, the spleen-targeted lipid nanoparticles include the following components in molar percentages:

[0033] 1) 5-10 mol% of steroid compound;

[0034] 2) ionizable lipids 40-60 mol%;

[0035] 3) 5-20 mol% phospholipids;

[0036] 4) structural lipids 15-70 mol%; and

[0037] 5) Polymer conjugated lipid 0.5-5 mol%.

[0038] In a preferred embodiment, the spleen-targeted lipid nanoparticles include the following components in molar percentages:

[0039] 1) 5-10 mol% of steroid compound;

[0040] 2) ionizable lipids 40-60 mol%;

[0041] 3) 5-15 mol% phospholipids;

[0042] 4) structural lipids 30-50 mol%; and

[0043] 5) Polymer conjugated lipid 0.5-3 mol%.

[0044] In a particularly preferred embodiment, the spleen-targeted lipid nanoparticles include the following components in molar percentages:

[0045] 1) steroid compound 5 mol%;

[0046] 2) ionizable lipids 50 mol%;

[0047] 3) phospholipids 10 mol%;

[0048] 4) structural lipids 33.5 mol%; and

[0049] 5) Polymer-conjugated lipid 1.5 mol%.

[0050] In one embodiment, the particle size distribution (PDI) of the lipid nanoparticles is no greater than 0.2, preferably no greater than 0.15.

[0051] In one embodiment, the ratio of the amount of the lipid nanoparticles distributed to the spleen to that distributed to the liver after administration is greater than 1, more preferably greater than 2, more preferably greater than 3, more preferably greater than 4, more preferably greater than 5, more preferably greater than 10, more preferably greater than 20, more preferably greater than 30, more preferably greater than 40, more preferably greater than 50, and more preferably greater than 100.

[0052] In one embodiment, the ratio of the lipid nanoparticles distributed to the spleen and the lungs after administration is greater than 1, more preferably greater than 2, more preferably greater than 3, more preferably greater than 4, more preferably greater than 5, more preferably greater than 10, more preferably greater than 20, more preferably greater than 30, more preferably greater than 40, more preferably greater than 50, and more preferably greater than 100.

[0053] In one aspect, provided herein is a lipid nanoparticle comprising an ionizable lipid and a steroid compound, wherein the steroid compound is a compound of formula (I):

[0054] or a stereoisomer, a mixture of stereoisomers, or a pharmaceutically acceptable salt thereof, wherein:

[0055] Is a single bond or a double bond;

[0056] Y is selected from -C(O)O-, -O-, -NH-, -SC(O)O-, -OC(O)NR 2a -、-NR 2a C(O)NR 2a -、-OC(O)S-、-OC(O)O-、-NR 2a C(O)O-, -OC(O)-, -SC(O)-, -C(O)S-, -NR 2a -、-C(O)NR 2a -、-NR 2a C(O)-、-NR 2a C(O)S-、-SC(O)NR 2a -, -C(O)-, -OC(S)-, -C(S)O-, -OC(S)NR 2a -、-NR 2a C(S)O- or -S(O) 0-2 -, and its left connection is towards the direction of V1;

[0057] Y1 is absent, -C(O)O-, -O-, -NH-, -SC(O)O-, -OC(O)NR 2a -、-NR 2a C(O)NR 2a -、-OC(O)S-、-OC(O)O-、-NR 2a C(O)O-, -OC(O)-, -SC(O)-, -C(O)S-, -NR 2a -、-C(O)NR 2a -、-NR 2a C(O)-、-NR 2a C(O)S-、-SC(O)NR 2a -, -C(O)-, -OC(S)-, -C(S)O-, -OC(S)NR 2a -、-NR 2a C(S)O- or -S(O) 0-2 -, and its left connection is towards the direction of V1;

[0058] R 2a Each example is independently selected from H, C 1-10 Alkyl, C 3-14 cycloalkyl or 3 to 14 membered heterocyclyl, and wherein the alkyl, cycloalkyl and heterocyclyl are independently optionally replaced by C 1-10 Alkyl, L 2b -OR 2b, Lb-SR 2b 、L 2b -NR 2b R' 2b 、L 2b N + R 2b R' 2b R” 2b 、L 2b -OC(O)R 2b or L 2b -C(O)OR 2b replace;

[0059] R 201 、R 202 、R 203 、R 204 Each example is independently selected from H, OH, alkyl, alkoxy, -L 2b -NR 21 R 22 、-L 2b -N + R 21 R 22 R 23 、L 2b -OC(O)R 21 or L 2b -C(O)OR 21 ;

[0060] m is an integer from 1 to 12;

[0061] m1 is an integer from 0 to 12;

[0062] V1 is NR 21 、N + R 21 R 22 R 23 , imidazole, pyridine, pyrrole, pyrrolidine, pyrazole, (iso)thiazole, (iso)oxazole, oxadiazine, oxazoline, dithioureaazole, (iso)triazole, tetrazole, pentazole, indole, indoline, pyrimidine, pyrazine, quinazoline, piperazine, piperidine, morpholine, pyran, quinoxaline, quinoline, quinolinone, (iso)quinoline, amidine, guanidine or urea;

[0063] M is absent or selected from one or more anions independently selected from chloride, bromide, iodide, sulfate, nitrate, perchlorate, formate, citrate, sulfonate, methanesulfonate, trifluoromethanesulfonate, trifluoroacetate, pyrosulfate, bisulfate, sulfite, bisulfite, phosphate, pyrophosphate, monohydrogen phosphate, dihydrogen phosphate, acetate, propionate, decanoate, octanoate, acrylate, isobutyrate, hexanoate, heptanoate, oxalate, malonate, succinate, fumarate, maleate, benzoate, phthalate, phenylacetate, lactate, glycolate, tartrate, mandelate, methanesulfonate, gluconate, aspartate, or glutamate;

[0064] R 21 、R 22 、R 23 Each instance of is independently H, optionally substituted C 1-8 Alkyl, -L 2b -OR 2b 、-L 2b -SR 2b 、-L 2b -NR 2b R' 2b R” 2b 、L 2b -OC(O)R 2b or L 2b -C(O)OR 2b ;

[0065] L 2b is absent or optionally substituted C 1-10 alkylene;

[0066] R 2b , R' 2b and R” 2b Each instance of is independently H or C 1-10 alkyl;

[0067] R z1 It is C 1-14 Alkyl or C 1-14 alkenyl, wherein the alkyl and alkenyl groups are optionally substituted; and

[0068] Provided that, when Y is -NHC(O)-, at least one of the following conditions is met:

[0069] (i) m is an integer from 5 to 12; or

[0070] (ii) V1 is not N(CH3)2.

[0071] In one embodiment, the present invention provides a lipid nanoparticle comprising an ionizable lipid and a steroid compound, wherein the steroid compound is a compound of formula (IP):

[0072] or a stereoisomer, a mixture of stereoisomers, or a pharmaceutically acceptable salt thereof, wherein:

[0073] Is a single bond or a double bond;

[0074] Y1 does not exist, -C(O)O-, -O-, -NH-, -SC(O)O-, -OC(O)NR 2a -、-NR 2a C(O)NR 2a -、-OC(O)S-、-OC(O)O-、-NR 2a C(O)O-, -OC(O)-, -SC(O)-, -C(O)S-,-NR 2a -,-C(O)NR 2a -,-NR 2a C(O)-,-NR 2a C(O)S-,-SC(O)NR 2a -,-C(O)-,-OC(S)-, -C(S)O-, -OC(S)NR 2a -、-NR 2a C(S)O-, -S(O) 0-2 -or -OPO3-, and wherein the leftward connection is in the direction of V1;

[0075] R 2a Each instance of is independently H, C 1-10 Alkyl, C 3-14 cycloalkyl or 3 to 14 membered heterocyclyl, and wherein the alkyl, cycloalkyl and heterocyclyl are independently optionally replaced by C 1-10 Alkyl, L 2b -OR 2b 、L b -SR 2b 、L 2b -NR 2b R' 2b 、L 2b N + R 2b R' 2b R” 2b 、L 2b -OC(O)R 2b or L 2b -C(O)OR 2b replace;

[0076] R201 、R 202、 R 203 、R 204 Each example is independently H, OH, alkyl, alkoxy, -L 2b -NR 21 R 22 、-L 2b -N + R 21 R 22 R 23 、L 2b -OC(O)R 21 or L 2b -C(O)OR 21 ;

[0077] R 205 Absence, cations, H, C 1-6 Alkyl, C 1-6 Halogenated alkyl, C 2-6 Alkenyl or C 2-6 alkynyl, and wherein alkyl, haloalkyl, alkenyl and alkynyl are optionally substituted;

[0078] m is an integer from 1 to 12;

[0079] m1 is an integer from 0 to 12;

[0080] V1 is NR 21 R 22 、N + R 21 R 22 R 23 , imidazole, pyridine, pyrrole, pyrrolidine, pyrazole, (iso)thiazole, (iso)oxazole, oxadiazine, oxazoline, dithioureaazole, (iso)triazole, tetrazole, pentazole, indole, indoline, pyrimidine, pyrazine, quinazoline, piperazine, piperidine, morpholine, pyran, quinoxaline, quinoline, quinolinone, (iso)quinoline, amidine, guanidine or urea;

[0081] M is absent or is one or more anions independently selected from chloride, bromide, iodide, sulfate, nitrate, perchlorate, formate, citrate, sulfonate, methanesulfonate, trifluoromethanesulfonate, trifluoroacetate, pyrosulfate, bisulfate, sulfite, bisulfite, phosphate, pyrophosphate, monohydrogen phosphate, dihydrogen phosphate, acetate, propionate, decanoate, octanoate, acrylate, isobutyrate, hexanoate, heptanoate, oxalate, malonate, succinate, fumarate, maleate, benzoate, phthalate, phenylacetate, lactate, glycolate, tartrate, mandelate, methanesulfonate, gluconate, aspartate, or glutamate;

[0082] R21 、R 22 、R 23 Each instance of is independently H, optionally substituted C 1-8 Alkyl, -L 2b -OR 2b 、-L 2b -SR 2b 、-L 2b -NR 2b R' 2b R” 2b 、L 2b -OC(O)R 2b or L 2b -C(O)OR 2b ;

[0083] L 2b is absent or optionally substituted C 1-10 alkylene;

[0084] R 2b , R' 2b and R” 2b Each instance of is independently H or C 1-10 alkyl;

[0085] R z1 It is C 1-14 Alkyl or C 1-14 alkenyl, wherein the alkyl and alkenyl groups are optionally substituted.

[0086] In one embodiment, Y is -C(O)O-, -OC(O)-, -O-, -NHC(O)O-, -OC(O)NH-, or -OC(O)O-, and wherein the leftward linkage is in the direction of V. In one embodiment, Y is absent, -C(O)O-, -OC(O)-, -O-, -S-, SO, S(O)2, -O(CO)O-, -NHC(O)-, or -C(O)NH-, and wherein the leftward linkage is in the direction of V.

[0087] In one embodiment, the steroid compound is a compound of Formula (I-A1), (I-A2), (I-A3), (I-A4), or (I-A5):

[0088] or a stereoisomer, a mixture of stereoisomers or a pharmaceutically acceptable salt thereof,

[0089] In one embodiment, the steroid compound is a compound of Formula (I-B1), (I-B2), (I-B3), (I-B4), (I-B5), (I-B6), (I-B7), or (I-B8):

[0090] or a stereoisomer, a mixture of stereoisomers, or a pharmaceutically acceptable salt thereof, wherein m3 is an integer from 1 to 11.

[0091] In one embodiment, R 201 、R 202 、R 203 and R 204 Each instance of is independently H, C 1-6 Alkyl, -NR 21 R 22 、-N + R 21 R 22 R 23 、-(CH2) 0-6 -OC(O)R 21 or -(CH2) 0-6 -C(O)OR 21 .

[0092] In one embodiment, R 201 、R 202 、R 203 and R 204 Each instance of is independently H, NH2, NH3 + , -CH2OC(O)CH3, -CH2C(O)OCH3, -OC(O)CH3 or -C(O)OCH3.

[0093] In one embodiment, m is an integer from 5 to 12.

[0094] In one embodiment, the present invention provides a lipid nanoparticle, wherein the steroid compound is a compound of formula (I-P1):

[0095] or a stereoisomer, a mixture of stereoisomers, or a pharmaceutically acceptable salt thereof, and wherein m3 is an integer from 1 to 11.

[0096] In one embodiment, V1 is NR 21 R 22 、N + R 21 R 22 R 23 , or an optionally substituted amidine, guanidine or urea radical. In one embodiment, R 21 、R 22 、R 23 are independently H, hydroxyl, C 1-3 Alkyl or C 1-3 In one embodiment, V1 is NH2, NH3 +、N(CH3)2、N + (CH3)3、N + CH3(CH2CH2OH)2、N + (CH3)2(CH2CH2OH), NHC(=N + H2)NH2, NHC(O)NH2, CH2C(=NH)NH2, N(CH3)(CH2CH2CO2CH3), N + (CH3)2(CH2CH2CO2CH3), N(CH3)(CH2CO2CH3) or N + (CH3)2(CH2CO2CH3). In one embodiment, V1 is N + CH3(CH2CH2OH)2、N + (CH3)2(CH2CH2OH) or N + (CH3)3.

[0097] In one embodiment, M is absent or is one or more of bromide, chloride, iodide, or trifluoroacetate.

[0098] In one embodiment, R z1 It is C 1-14 Alkyl or C 1-14 Alkenyl, wherein the alkyl and alkenyl groups are optionally substituted with one or more hydroxy, oxo, nitro, cyano, halogen, C 1-6 Alkoxy, C2-C6 alkenyl, C2-C6 alkynyl, C3-C8 cycloalkyl, C6-C 10 In one embodiment, R z1 It is C 6-12 Alkyl or C 6-12 In one embodiment, R z1 yes

[0099] In another embodiment, the steroid compound is a compound of formula (II-A):

[0100] or a stereoisomer, a mixture of stereoisomers, or a pharmaceutically acceptable salt thereof, wherein:

[0101] Each are independently a single bond or a double bond;

[0102] Y is -C(O)O-, -O-, -NH-, -SC(O)O-, -OC(O)NR 2a-、-NR 2a C(O)NR 2a -、-OC(O)S-、-OC(O)O-、-NR 2a C(O)O-, -OC(O)-, -SC(O)-, -C(O)S-, -NR 2a -、-C(O)NR 2a -、-NR 2a C(O)-、-NR 2a C(O)S-、-SC(O)NR 2a -, -C(O)-, -OC(S)-, -C(S)O-, -OC(S)NR 2a -、-NR 2a C(S)O- or -S(O) 0-2 -, and wherein the leftward connection is in the direction of V1;

[0103] Y1 is absent, -C(O)O-, -O-, -NH-, -SC(O)O-, -OC(O)NR 2a -、-NR 2a C(O)NR 2a -、-OC(O)S-、-OC(O)O-、-NR 2a C(O)O-, -OC(O)-, -SC(O)-, -C(O)S-, -NR 2a -、-C(O)NR 2a -、-NR 2a C(O)-、-NR 2a C(O)S-、-SC(O)NR 2a -, -C(O)-, -OC(S)-, -C(S)O-, -OC(S)NR 2a -、-NR 2a C(S)O- or -S(O) 0-2 -, and wherein the leftward connection is in the direction of V1;

[0104] R 2a Each instance of is independently H, C 1-10 Alkyl, C 3-14 cycloalkyl or 3 to 14 membered heterocyclyl, and wherein the alkyl, cycloalkyl and heterocyclyl are independently optionally replaced by C 1-10 Alkyl, L 2b -OR 2b 、L b -SR 2b 、L 2b -NR 2b R' 2b 、L 2b N + R 2bR' 2b R” 2b 、L 2b -OC(O)R 2b or L 2b -C(O)OR 2b replace;

[0105] R 201 、R 202、 R 203 、R 204 Each example is independently H, OH, alkyl, alkoxy, -L 2b -NR 21 R 22 or -L 2b -N + R 21 R 22 R 23 、L 2b -OC(O)R 21 or L 2b -C(O)OR 21 ;

[0106] m is an integer from 3 to 12;

[0107] m1 is an integer from 0 to 12;

[0108] V1 is NR 21 、N + R 21 R 22 R 23 , imidazole, pyridine, pyrrole, pyrrolidine, pyrazole, (iso)thiazole, (iso)oxazole, oxadiazine, oxazoline, dithioureaazole, (iso)triazole, tetrazole, pentazole, indole, indoline, pyrimidine, pyrazine, quinazoline, piperazine, piperidine, morpholine, pyran, quinoxaline, quinoline, quinolinone, (iso)quinoline, amidine, guanidine or urea;

[0109] M is absent or is one or more anions independently selected from chloride, bromide, iodide, sulfate, nitrate, perchlorate, formate, citrate, sulfonate, methanesulfonate, trifluoromethanesulfonate, trifluoroacetate, pyrosulfate, bisulfate, sulfite, bisulfite, phosphate, pyrophosphate, monohydrogen phosphate, dihydrogen phosphate, acetate, propionate, decanoate, octanoate, acrylate, isobutyrate, hexanoate, heptanoate, oxalate, malonate, succinate, fumarate, maleate, benzoate, phthalate, phenylacetate, lactate, glycolate, tartrate, mandelate, methanesulfonate, gluconate, aspartate, or glutamate;

[0110] R 21 、R 22 、R23 are each independently H, optionally substituted C 1-8 Alkyl, -L 2b -OR 2b 、-L 2b -SR 2b 、-L 2b -NR 2b R' 2b R” 2b 、L 2b -OC(O)R 2b or L 2b -C(O)OR 2b ;

[0111] L 2b is absent or optionally substituted C 1-10 alkylene;

[0112] R 2b , R' 2b and R” 2b Each of is independently H or C 1-10 alkyl; and

[0113] R z1 It is C 1-14 Alkyl or C 1-14 alkenyl, wherein the alkyl and alkenyl groups are optionally substituted; and with the proviso that the compound is not:

[0114] In one embodiment, the steroid compound is of formula (II-B):

[0115] or a stereoisomer, a mixture of stereoisomers, or a pharmaceutically acceptable salt thereof, wherein:

[0116] Y is -C(O)O-, -O-, -NH-, -SC(O)O-, -OC(O)NR 2a -、-NR 2a C(O)NR 2a -、-OC(O)S-、-OC(O)O-、-NR 2a C(O)O-, -OC(O)-, -SC(O)-, -C(O)S-, -NR 2a -、-C(O)NR 2a -、-NR 2a C(O)-、-NR 2a C(O)S-、-SC(O)NR 2a -, -C(O)-, -OC(S)-, -C(S)O-, -OC(S)NR 2a -、-NR2a C(S)O- or -S(O) 0-2 -, and wherein the leftward connection is in the direction of V1;

[0117] Y1 is absent, -C(O)O-, -O-, -NH-, -SC(O)O-, -OC(O)NR 2a -、-NR 2a C(O)NR 2a -、-OC(O)S-、-OC(O)O-、-NR 2a C(O)O-, -OC(O)-, -SC(O)-, -C(O)S-, -NR 2a -、-C(O)NR 2a -、-NR 2a C(O)-、-NR 2a C(O)S-、-SC(O)NR 2a -, -C(O)-, -OC(S)-, -C(S)O-, -OC(S)NR 2a -、-NR 2a C(S)O- or -S(O) 0-2 -, and wherein the leftward connection is in the direction of V1;

[0118] R 2a Each instance of is independently H, C 1-10 Alkyl, C 3-14 cycloalkyl or 3 to 14 membered heterocyclyl, and wherein the alkyl, cycloalkyl and heterocyclyl are independently optionally replaced by C 1-10 Alkyl, L 2b -OR 2b 、L b -SR 2b 、L 2b -NR 2b R' 2b 、L 2b N + R 2b R' 2b R” 2b 、L 2b -OC(O)R 2b or L 2b -C(O)OR 2b replace;

[0119] R 201 、R 202、 R 203 、R 204 Each example is independently H, OH, alkyl, alkoxy, -L 2b -NR 21 R 22 or -L2b -N + R 21 R 22 R 23 、L 2b -OC(O)R 21 or L 2b -C(O)OR 21 ;

[0120] m is an integer from 5 to 12;

[0121] m1 is an integer from 0 to 12;

[0122] V1 is NR 21 R 22 、N + R 21 R 22 R 23 , imidazole, pyridine, pyrrole, pyrrolidine, pyrazole, (iso)thiazole, (iso)oxazole, oxadiazine, oxazoline, dithioureaazole, (iso)triazole, tetrazole, pentazole, indole, indoline, pyrimidine, pyrazine, quinazoline, piperazine, piperidine, morpholine, pyran, quinoxaline, quinoline, quinolinone, (iso)quinoline, amidine, guanidine or urea;

[0123] M is absent or is one or more anions independently selected from chloride, bromide, iodide, sulfate, nitrate, perchlorate, formate, citrate, sulfonate, methanesulfonate, trifluoromethanesulfonate, trifluoroacetate, pyrosulfate, bisulfate, sulfite, bisulfite, phosphate, pyrophosphate, monohydrogen phosphate, dihydrogen phosphate, acetate, propionate, decanoate, octanoate, acrylate, isobutyrate, hexanoate, heptanoate, oxalate, malonate, succinate, fumarate, maleate, benzoate, phthalate, phenylacetate, lactate, glycolate, tartrate, mandelate, methanesulfonate, gluconate, aspartate, or glutamate;

[0124] R 21 、R 22 、R 23 are each independently H, optionally substituted C 1-8 Alkyl, -L 2b -OR 2b 、-L 2b -SR 2b 、-L 2b -NR 2b R' 2b R” 2b 、L 2b -OC(O)R 2b or L 2b -C(O)OR 2b ;

[0125] L 2b is absent or optionally substituted C 1-10 alkylene;

[0126] R 2b , R' 2b and R” 2b Each of is independently H or C 1-10 alkyl.

[0127] In one embodiment of formula (II-B), Y1 is -C(O)O-, -OC(O)-, -O-, -S-, -NR 2a -、-OC(O)NR 2a -or-NR 2a C(O)-.

[0128] In another embodiment, the steroid compound is a compound of formula (II-C1):

[0129] or a stereoisomer, a mixture of stereoisomers, or a pharmaceutically acceptable salt thereof, wherein:

[0130] R z1 yes

[0131] R 2a is H, optionally substituted C 1-8 Alkyl, -L 2b -OR 2b 、L 2b -OC(O)R 2b or L 2b -C(O)OR 2b , preferably H, -CH3, -CH2C(O)OCH3 or -CH2C(O)OH;

[0132] V1 is NR 21 R 22 or N + R 21 R 22 R 23 ; where R 21 、R 22 、R 23 are each independently H, optionally substituted C 1-8 Alkyl, -L 2b -OR 2b 、-L 2b -SR 2b 、-L 2b -NR 2b R' 2b R”2b 、L 2b -OC(O)R 2b or L 2b -C(O)OR 2b ; and R 21 and R 22 Not simultaneously hydrogen;

[0133] L 2b is absent or optionally substituted C 1-10 alkylene;

[0134] R 2b , R' 2b and R” 2b Each of is independently H or C 1-10 alkyl;

[0135] M is absent or is one or more anions independently selected from chloride, bromide, iodide, sulfate, nitrate, perchlorate, formate, citrate, sulfonate, methanesulfonate, trifluoromethanesulfonate, trifluoroacetate, pyrosulfate, bisulfate, sulfite, bisulfite, phosphate, pyrophosphate, monohydrogen phosphate, dihydrogen phosphate, acetate, propionate, decanoate, octanoate, acrylate, isobutyrate, hexanoate, heptanoate, oxalate, malonate, succinate, fumarate, maleate, benzoate, phthalate, phenylacetate, lactate, glycolate, tartrate, mandelate, methanesulfonate, gluconate, aspartate, or glutamate;

[0136] m is an integer from 1 to 10, for example, 1, 2, 3, 4, 5, 6, 7, 8, 9 or 10.

[0137] In another embodiment, the steroid compound is a compound of formula (II-C2):

[0138] or a stereoisomer, a mixture of stereoisomers, or a pharmaceutically acceptable salt thereof, wherein:

[0139] R z1 yes

[0140] V1 is NR 21 R 22 or N + R 21 R 22 R 23 ; where R 21 、R 22 、R 23 are each independently H, optionally substituted C 1-8 Alkyl, -L2b -OR 2b 、-L 2b -SR 2b 、-L 2b -NR 2b R' 2b R” 2b 、L 2b -OC(O)R 2b or L 2b -C(O)OR 2b ; and R 21 and R 22 Not simultaneously hydrogen;

[0141] L 2b is absent or optionally substituted C 1-10 alkylene;

[0142] R 2b , R' 2b and R” 2b Each of is independently H or C 1-10 alkyl;

[0143] M is absent or is one or more anions independently selected from chloride, bromide, iodide, sulfate, nitrate, perchlorate, formate, citrate, sulfonate, methanesulfonate, trifluoromethanesulfonate, trifluoroacetate, pyrosulfate, bisulfate, sulfite, bisulfite, phosphate, pyrophosphate, monohydrogen phosphate, dihydrogen phosphate, acetate, propionate, decanoate, octanoate, acrylate, isobutyrate, hexanoate, heptanoate, oxalate, malonate, succinate, fumarate, maleate, benzoate, phthalate, phenylacetate, lactate, glycolate, tartrate, mandelate, methanesulfonate, gluconate, aspartate, or glutamate;

[0144] m is an integer from 1 to 10, for example, 1, 2, 3, 4, 5, 6, 7, 8, 9 or 10.

[0145] In another aspect, provided herein is a lipid nanoparticle comprising an ionizable lipid and a steroid compound, wherein the steroid compound is:

[0146] or a stereoisomer, a mixture of stereoisomers or a pharmaceutically acceptable salt thereof.

[0147] In another aspect, provided herein is a lipid nanoparticle for delivering or expressing a payload in the spleen of a subject, wherein the lipid nanoparticle is administered to the subject by a systemic administration route, wherein the lipid nanoparticle comprises an ionizable lipid and a steroid compound, and wherein the steroid compound comprises one or more cationic groups.

[0148] In one embodiment, the systemic route of administration is intravenous, intraarterial, or intraperitoneal.

[0149] In one embodiment, the cationic group is a tertiary amine, a quaternary ammonium, a primary amine, an ammonium, an imidazole, a pyridine, a pyrrole, a pyrrolidine, a pyrazole, an isothiazole, an isoxazole, an oxadiazine, an oxazoline, a dithioureaazole, an isotriazole, a tetrazole, a pentazole, an indole, a dihydroindole, an isoxazole, a pyrimidine, a pyrazine, a quinazoline, a piperazine, a piperidine, a morpholine, a pyran, a quinoxaline, a quinoline, a quinolinone, an isoquinoline, an amidine, a guanidine, or a urea.

[0150] In one embodiment, the steroid compound comprises a Z1 moiety, and wherein said Z1 is:

[0151] or a stereoisomer or mixture of stereoisomers thereof, wherein one or more ring C—C bonds in Z1 are independently optionally replaced by C═C bonds where valence permits, wherein Z1 is optionally replaced by one or more rings selected from hydroxy, halogen and C 1-14 Alkyl or C 1-14 wherein the alkyl group is optionally substituted with one or more hydroxyl, nitro, cyano, halogen, C1-6 alkoxy, C2-C6 alkenyl, C2-C6 alkynyl, C3-C8 cycloalkyl, C6-C 10 In one embodiment, the Z1 moiety is substituted with an aryl, a 5- to 10-membered heteroaryl, or a 3- to 8-membered heterocyclyl, and wherein the connection in Z1 is toward the rest of the steroid compound. In one embodiment, the Z1 moiety is a monovalent group of cholesterol, cholestane, campesterol, sitosterol, sitostanol, β-sitosterol, stigmasterol, stigmasterol, brassicasterol, avenasterol, ergosterol, ursolic acid, tocopherol, or a stereoisomer thereof, a mixture of stereoisomers, or a pharmaceutically acceptable salt thereof.

[0152] In one embodiment, the steroid compound is a compound of formula (I):

[0153] or a stereoisomer, a mixture of stereoisomers, or a pharmaceutically acceptable salt thereof, wherein:

[0154] Is a single bond or a double bond;

[0155] Y is -C(O)O-, -O-, -NH-, -SC(O)O-, -OC(O)NR 2a -、-NR 2a C(O)NR 2a -、-OC(O)S-、-OC(O)O-、-NR 2a C(O)O-, -OC(O)-, -SC(O)-, -C(O)S-, -NR 2a -、-C(O)NR 2a -、-NR 2a C(O)-、-NR 2a C(O)S-、-SC(O)NR 2a -, -C(O)-, -OC(S)-, -C(S)O-, -OC(S)NR 2a -、-NR 2a C(S)O- or -S(O) 0-2 -, and wherein the leftward connection is in the direction of V1;

[0156] Y1 is absent, -C(O)O-, -O-, -NH-, -SC(O)O-, -OC(O)NR 2a -、-NR 2a C(O)NR 2a -、-OC(O)S-、-OC(O)O-、-NR 2a C(O)O-, -OC(O)-, -SC(O)-, -C(O)S-, -NR 2a -、-C(O)NR 2a -、-NR 2a C(O)-、-NR 2a C(O)S-、-SC(O)NR 2a -, -C(O)-, -OC(S)-, -C(S)O-, -OC(S)NR 2a -、-NR 2a C(S)O- or -S(O) 0-2 -, and wherein the leftward connection is in the direction of V1;

[0157] R 2a Each instance of is independently H, C 1-10 Alkyl, C 3-14 cycloalkyl or 3 to 14 membered heterocyclyl, and wherein the alkyl, cycloalkyl and heterocyclyl are independently optionally replaced by C 1-10 Alkyl, L 2b -OR 2b , Lb-SR 2b 、L 2b -NR 2b R' 2b 、L2b N + R 2b R' 2b R” 2b 、L 2b -OC(O)R 2b or L 2b -C(O)OR 2b replace;

[0158] R 201 、R 202 、R 203 、R 204 Each example is independently H, OH, alkyl, alkoxy, -L 2b -NR 21 R 22 、-L 2b -N + R 21 R 22 R 23 、L 2b -OC(O)R 21 or L 2b -C(O)OR 21 ;

[0159] m is an integer from 1 to 12;

[0160] m1 is an integer from 0 to 12;

[0161] V1 is NR 21 R 22 、N + R 21 R 22 R 23 , imidazole, pyridine, pyrrole, pyrrolidine, pyrazole, isothiazole, isoxazole, oxadiazine, oxazoline, dithioureaazole, isotriazole, tetrazole, pentazole, indole, indoline, isoxazole, pyrimidine, pyrazine, quinazoline, piperazine, piperidine, morpholine, pyran, quinoxaline, quinoline, quinolinone, isoquinoline, amidine, guanidine or urea;

[0162] M is absent or is one or more anions independently selected from chloride, bromide, iodide, sulfate, nitrate, perchlorate, formate, citrate, sulfonate, methanesulfonate, trifluoromethanesulfonate, trifluoroacetate, pyrosulfate, bisulfate, sulfite, bisulfite, phosphate, pyrophosphate, monohydrogen phosphate, dihydrogen phosphate, acetate, propionate, decanoate, octanoate, acrylate, isobutyrate, hexanoate, heptanoate, oxalate, malonate, succinate, fumarate, maleate, benzoate, phthalate, phenylacetate, lactate, glycolate, tartrate, mandelate, methanesulfonate, gluconate, aspartate, or glutamate;

[0163] R 21 、R 22 、R 23 Each instance of is independently H, optionally substituted C 1-8 Alkyl, -L 2b -OR 2b 、-L 2b -SR 2b 、-L 2b -NR 2b R' 2b R” 2b 、L 2b -OC(O)R 2b or L 2b -C(O)OR 2b ;

[0164] L 2b is absent or optionally substituted C 1-10 alkylene;

[0165] R 2b , R' 2b and R” 2b Each instance of is independently H or C 1-10 alkyl; and

[0166] R z1 It is C 1-14 Alkyl or C 1-14 alkenyl, wherein the alkyl and alkenyl groups are optionally substituted.

[0167] In one embodiment, the amount of the steroid compound accounts for about 15 mol % to about 80 mol %, about 20 mol % to about 50 mol %, about 20 mol % to about 40 mol % or about 15 mol % to about 30 mol % of the total lipid in the lipid nanoparticle. In one embodiment, the amount of the steroid compound accounts for about 20 mol % to about 30 mol % of the total lipid in the lipid nanoparticle.

[0168] In one embodiment, the ionizable lipid comprises at least one ionizable head group and at least one biodegradable hydrophobic chain, wherein the ionizable lipid has a logP of at least about 10.1 and a pKa of about 4 to about 11; and the biodegradable hydrophobic chain has the structural formula -R 16 -M0-R 17 ; where R 16 It is C 4-14 Alkylene, C 4-14 Alkenylene or C 4-14 Alkynylidene; R 17 It is C 6-20alkyl; and M0 is a biodegradable group, and wherein the alkylene, alkenylene, alkynylene and alkyl are optionally substituted.

[0169] In one embodiment, M0 is an ester group, an amide group, a thioester group, a carbonate group, a carbamate group, a carbamoylthioester group, a urea group, or a disulfide group.

[0170] In one embodiment, the ionizable lipid is a compound of formula (III):

[0171] or a stereoisomer, mixture of stereoisomers, tautomer, isotopomer or pharmaceutically acceptable salt thereof, wherein:

[0172] Each instance of R3 and R4 is independently C 1-10 Alkyl, C 2-10 Alkenyl, C 2-10 Alkynyl, C3-C 14 Cycloalkyl, 3 to 14 membered heterocyclic group, (C 1-10 Alkylene)-(C3-C 14 Cycloalkyl) or (C 1-10 alkylene)-(3 to 14 membered heterocyclyl), or R3 and R4 together with the nitrogen atom to which they are attached form a 3 to 14 membered heterocyclyl, or R4 and one R0' together with the atom to which they are attached form a 4 to 10 membered heterocyclyl or a 5 to 10 membered heteroaryl, and wherein said alkyl, alkenyl, alkynyl, cycloalkyl, heterocyclyl, heteroaryl and alkylene are independently optionally substituted with one or more R*;

[0173] Each instance of R0' is independently optionally substituted methylene, or wherein two R0' taken together with the carbon to which they are attached form a 3- to 8-membered cycloalkylene, and wherein said cycloalkylene is optionally substituted with one or more R**;

[0174] k is 0, 1, 2, 3, 4, 5, or 6;

[0175] j is 0 or 1;

[0176] The dotted line connecting Q and W does not exist or is a chemical bond;

[0177] W is C, CH or N, provided that when W is N, j is 0 and the dotted bond connecting Q and W is absent;

[0178] G5 does not exist, C 1-24 Alkylene, C 2-24 Alkenylene, C3-C8 cycloalkylene, C3-C8 cycloalkenylene, wherein the alkylene, alkenylene, cycloalkylene, cycloalkenylene are optionally substituted with one or more R**;

[0179] G1, G2, G3 and G4 are each independently absent, C 1-13 Alkylene, C 2-13 Alkenylene or C 2-13 Alkyne, wherein the alkylene, alkenylene and alkynylene groups are optionally substituted by one or more R s replace;

[0180] The total length of G1 and G2 is 3, 4, 5, 6, 7, 8, 9, 10, 11, 12 or 13 carbon atoms;

[0181] The total length of G3 and G4 is 3, 4, 5, 6, 7, 8, 9, 10, 11, 12 or 13 carbon atoms;

[0182] R5, R6, R7 and R8 are each independently hydrogen or optionally substituted C 1-8 alkyl;

[0183] When the dashed line connecting Q and W is absent, Q is absent or selected from -C(O)O-, -O-, -NH-, -SC(O)O-, -OC(O)NR b -、-NR b C(O)NR b -、-OC(O)S-、-OC(O)O-、-NR b C(O)O-, -OC(O)-, -SC(O)-, -C(O)S-, -NR b -、-C(O)NR b -、-NR b C(O)-、-NR b C(O)S-、-SC(O)NR b -, -C(O)-, -OC(S)-, -C(S)O-, -OC(S)NR b -、-NR b C(S)O-, -SS-, or -S(O) 0-2 -, where the direction of W is connected to the right;

[0184] When the dashed line connecting Q and W is a chemical bond, G5 is absent, and Q and W form a 5- to 10-membered monocyclic or fused ring, optionally wherein the ring is substituted with one or more R**;

[0185] M1 and M2 are each independently absent or selected from -C(O)O-, -O-, -SC(O)O-, -OC(O)NR a -、-NR a C(O)NR a -、-OC(O)S-、-OC(O)O-、-NR aC(O)O-, -OC(O)-, -SC(O)-, -C(O)S-, -NR a -、-C(O)NR a -、-NR a C(O)-、-NR a C(O)S-、-SC(O)NR a -, -C(O)-, -OC(S)-, -C(S)O-, -OC(S)NR a -、-NR a C(S)O-, -SS-, or -S(O) 0-2 -;

[0186] Each instance of R0 is independently -(CRR')-;

[0187] Each instance of R and R' is independently H, C 1-20 Alkyl, L a -OR a 、-L a -SR a or -L a -NR a R' a , or R and R' together with the carbon to which they are attached form C 3-8 cycloalkylene;

[0188] Q3 and Q4 are each independently H, -(CRR')-, C 6-10 aryl or steroid groups;

[0189] Each instance of A1, A2, A3, A4 is independently -(CR 18 R 18 -CR 18 =CR 18 )-or-(CR 18 R 18 -C≡C)-;

[0190] R 18 Each example of C is independently 1-20 alkyl;

[0191] Each instance of N1 and N2 is independently a biodegradable group;

[0192] Z does not exist, C 1-10 Alkylene or -OP(O)(OH)-O-;

[0193] The dashed line between Z-Q3 or Z-Q4 independently does not exist or is a chemical bond, provided that when Z is absent, both dashed lines do not exist;

[0194] m, n, q, r, u, v, y and z are each independently selected from 0, 1, 2, 3, 4, 5, 6, 7, 8, 9 or 10;

[0195] o, p, w and x are each independently selected from 0, 1 or 2;

[0196] s and t are each independently selected from 0 or 1;

[0197] The number of carbon atoms between G1 and Q3 or between G3 and Q4 is 8 to 30;

[0198] L a and L e Each independently absent or selected from C 1-20 alkylene;

[0199] L b and L f Each independently absent or selected from C 1-10 alkylene;

[0200] L c Not present or selected from C 1-8 alkylene;

[0201] L d Not present or selected from C 1-14 alkylene;

[0202] R* is hydroxyl, halogen, C 1-10 Alkyl, C 1-10 Halogenated alkyl, C 1-10 Hydroxyalkyl, -L b -OR b 、-L b -SR b or -L b -NR b R' b ;

[0203] R** is hydroxyl, halogen, C 1-8 Alkyl, C 1-8 Halogenated alkyl, C 1-8 Hydroxyalkyl, -L b -OR b 、-L b -SR b or -L b -NR b R' b ;

[0204] R s It is C 1-14 Alkyl, -L b -OR b 、-L b -SR bor -L b -NR b R' b ;

[0205] R a and R' a Each independently is H, C 1-20 Alkyl, C 3-14 Cycloalkyl or 3 to 14 membered heterocyclyl, wherein the alkyl, cycloalkyl and heterocyclyl are optionally substituted by one or more selected from hydroxy, halogen, C 1-20 Alkyl, -L e -OR e 、-L e -SR e or -L e -NR e R' e The group substitution;

[0206] R b and R' b Each independently is H, C 1-10 Alkyl, C 3-14 Cycloalkyl or 3 to 14 membered heterocyclyl, wherein the alkyl, cycloalkyl and heterocyclyl are optionally substituted by one or more selected from hydroxy, halogen, C 1-10 Alkyl, -L f -OR f 、-L f -SR f or -L f -NR f R' f The group substitution;

[0207] R c and R' c are each independently hydrogen or C 1-8 alkyl;

[0208] R d and R' d are each independently hydrogen or C 1-14 alkyl;

[0209] R e and R' e are each independently hydrogen or C 1-20 alkyl; and

[0210] R f and RR' f are each independently hydrogen or C 1-10 alkyl.

[0211] In one embodiment, the ionizable lipid is a compound of formula (IV'):

[0212] or a stereoisomer, a mixture of stereoisomers or a pharmaceutically acceptable salt thereof, wherein

[0213] j is 0 or 1;

[0214] W is CH or N, provided that when W is N, j is 0;

[0215] k is 0, 1, 2, 3, 4, 5, 6, 7, or 8;

[0216] Each instance of R0' is independently optionally substituted methylene, or wherein two R0' together with the carbon to which they are attached form a 3- to 8-membered cycloalkylene, and wherein the cycloalkylene is optionally substituted with one or more R**;

[0217] M1 and M2 are each independently -C(O)O-, -O-, -SC(O)O-, -OC(O)NR a -、-NR a C(O)NR a -、-OC(O)S-、-OC(O)O-、-NR a C(O)O-, -OC(O)-, -SC(O)-, -C(O)S-, -NR a -、-C(O)NR a -、-NR a C(O)-、-NR a C(O)S-、-SC(O)NR a -, -C(O)-, -OC(S)-, -C(S)O-, -OC(S)NR a -、-NR a C(S)O-, -SS-, or -S(O) 0-2 -;

[0218] Q does not exist, -C(O)O-, -O-, -NH-, -SC(O)O-, -OC(O)NR b -、-NR b C(O)NR b -、-OC(O)S-、-OC(O)O-、-NR b C(O)O-, -OC(O)-, -SC(O)-, -C(O)S-, -NR b -、-C(O)NR b -、-NR b C(O)-、-NR b C(O)S-、-SC(O)NR b -, -C(O)-, -OC(S)-, -C(S)O-, -OC(S)NR b -、-NRb C(S)O-, -SS-, or -S(O) 0-2 -;

[0219] R a and R b Each independently is H, C 1-20 Alkyl, C 3-14 Cycloalkyl or 3 to 14 membered heterocyclyl, wherein the alkyl, cycloalkyl and heterocyclyl are optionally substituted by one or more selected from hydroxy, halogen, C 1-20 Alkyl, -Le-ORe, -Le-SRe or -Le-NReR'e;

[0220] G5 is absent or optionally substituted C 1-8 alkylene;

[0221] R1 and R2 are each independently C 4-20 Alkyl, C 4-20 Alkenyl or C 4-20 Alkynyl, wherein one or more methylene units in R1 and R2 are independently optionally replaced by -NH- or -N(C 1-20 alkyl), wherein the alkyl, alkenyl and alkynyl groups are optionally substituted;

[0222] Each instance of R3 and R4 is independently H, C 1-10 Alkyl, C 2-10 Alkenyl, C 2-10 Alkynyl, C3-C 14 cycloalkyl or 3 to 14 membered heterocyclyl, or R3 and R4 together with the nitrogen atom to which they are attached form a 3 to 14 membered heterocyclyl, or R4 and one R0' together with the atom to which they are attached form a 4 to 10 membered heterocyclyl; and wherein said alkyl, alkenyl, alkynyl, cycloalkyl, heterocyclyl is optionally substituted with one or more R*; and

[0223] R5, R6, R7 and R8 are each independently hydrogen or an optionally substituted C 1-8 alkyl.

[0224] In one embodiment, the ionizable lipid is a compound of formula (V'), (VI') or (VII'):

[0225] or a stereoisomer, a mixture of stereoisomers or a pharmaceutically acceptable salt thereof, wherein

[0226] k is 0, 1, 2, 3, 4, or 5;

[0227] L1 and L2 are each independently -(CRR')2-, -CH=CH-, -C≡C- or -NR"-;

[0228] One of L3 and L5 is -(CR s R s ')2-, -CH=CH- or -C≡C-, and the other is absent;

[0229] One of L4 and L6 is -(CR s R s ')2-, -CH=CH- or -C≡C-, and the other is absent;

[0230] G 1a , G 1b , G 2a , G 2b , G 3a , G 3b , G 4a and G 4b Each of which is independently absent or optionally substituted C1-7 alkylene;

[0231] G 1a , G 1b , G 2a and G 2b The total length of is 1, 2, 3, 4, 5, 6 or 7 carbon atoms;

[0232] G 3a , G 3b , G 4a and G 4b The total length of is 1, 2, 3, 4, 5, 6 or 7 carbon atoms;

[0233] R5, R6, R7 and R8 are each independently hydrogen or an optionally substituted C 1-6 alkyl;

[0234] G7, G8, G9 and G 10 Each independently absent or optionally substituted C 1-12 Alkylene, provided that the total length of G7 and G8 is 4, 5, 6, 7, 8, 9, 10, 11 or 12 carbon atoms, and G9 and G 10 has a total length of 4, 5, 6, 7, 8, 9, 10, 11 or 12 carbon atoms;

[0235] R s and R s 'Each independently is H, C 1-10 Alkyl, -L d -OR d or -L d -NR d R' d ;

[0236] R' is H, C 1-14 Alkyl, -L a -OR a or -L a -NR a R' a ;

[0237] R" is H or C 1-14 alkyl;

[0238] L d Does not exist or C 1-10 alkylene;

[0239] L a Does not exist or C 1-14 alkylene;

[0240] R a and R' a Each independently is H, C 1-14 alkyl, C3-10 cycloalkyl or 3 to 10 membered heterocyclyl, and wherein the alkyl, cycloalkyl and heterocyclyl are optionally substituted;

[0241] R d and R' d are each independently H or C 1-10 alkyl;

[0242] a' and b are each independently 0, 1, 3, 4, 5, provided that at least one of a' and b is not 0;

[0243] g is 0, 1, 2, 3, 4, or 5;

[0244] a'+g is 0, 1, 2, 3, 4 or 5; and

[0245] c, d, e and f are each independently 0, 1, 2, 3, 4, 5, 6 or 7, provided that c+d is an integer between 2 and 9, and e+f is an integer between 2 and 9.

[0246] In one embodiment, the ionizable lipid is a compound of formula (VIII):

[0247] or a stereoisomer, a mixture of stereoisomers or a pharmaceutically acceptable salt thereof.

[0248] In one embodiment, R5, R6, R7 and R8 are each independently hydrogen or methyl.

[0249] In one embodiment, G1, G2, G3 and G4 are each independently absent or C 1-8 Alkylene.

[0250] In one embodiment, the ionizable lipid is a compound of formula (IX):

[0251] or a stereoisomer, a mixture of stereoisomers or a pharmaceutically acceptable salt thereof, wherein

[0252] G1 and G2 are each independently a bond, C 1-13 Straight chain alkylene, C 2-13 Straight chain alkenylene or C 2-13 Straight chain alkynylene, wherein the alkylene, alkenylene and alkynylene groups are optionally substituted by one or more R G1 replace;

[0253] The total length of G1 and G2 is 3, 4, 5, 6, 7, 8, 9, 10, 11, 12 or 13 carbon atoms;

[0254] R G1 In each case, independently H, C 1-14 Alkyl, -L a -OR a 、-L a -SR a or -L a -NR a R' a ;

[0255] G3 is C 4-14 Straight chain alkylene, C 4-14 Straight chain alkenylene or C 4-14 Straight chain alkynylene, wherein the alkylene, alkenylene and alkynylene groups are optionally substituted by one or more R G3 replace;

[0256] R G3 In each case, independently H, -L a -OR a 、-L a -SR a or -L a -NR a R' a ;

[0257] L a independently a bond or C 1-14 alkylene;

[0258] R a and R' a Each independently is H, C 1-14 Alkyl, C 3-14 Cycloalkyl or 3-14 membered heterocyclic group;

[0259] G4 is the key, C 1-6 Alkylene, C2-6 Alkenylene or C 2-6 Alkyne, wherein the alkylene, alkenylene and alkynylene groups are optionally substituted by one or more R G4 replace;

[0260] R G4 In each case, independently H, C 1-6 Alkyl, -L b -OR b 、-L b -SR b or -L b -NR b R' b ;

[0261] L b independently a bond or C 1-6 alkylene;

[0262] R b and R' b Independently H, C 1-6 Alkyl, C 3-10 Cycloalkyl or 3-10 membered heterocyclic group;

[0263] Or two R G4 Together with the carbon atom to which they are attached, they form C 3-14 Cycloalkylene or 3-14 membered heterocyclylene, wherein the cycloalkylene and heterocyclylene are optionally substituted by one or more R 4g replace;

[0264] R 4g is independently H, halogen, cyano, C 1-8 Alkyl, C 1-8 Halogenated alkyl, -L e -OR e 、-L e -SR e or -L e -NR e R' e ;

[0265] L e independently a bond or C 1-8 alkylene;

[0266] R e and R' e Independently H, C 1-8 Alkyl, C 3-14 Cycloalkyl or 3-14 membered heterocyclic group;

[0267] M1 and M2 are independently -C(O)O-, -OC(O)-, -O-, -SC(O)O-, -OC(O)NR-, -NRC(O)NR-, -OC(O)S-, -OC(O)O-, -NRC(O)O-, -SC(O)-, -C(O )S-, -NR-, -C(O)NR-, -NRC(O)-, -NRC(O)S-, -SC(O)NR-, -C(O)-, -OC(S)-, -C(S)O-, -OC(S)NR-, -NRC(S)O-, -SS- or -S(O) 0-2 -;

[0268] Q is a bond, -C(O)O-, -O-, -SC(O)O-, -OC(O)NR f -、-NR f C(O)NR f -、-OC(O)S-、-OC(O)O-、-NR f C(O)O-, -OC(O)-, -SC(O)-, -C(O)S-, -NR f -、-C(O)NR f -、-NR f C(O)-、-NR f C(O)S-、-SC(O)NR f -, -C(O)-, -OC(S)-, -C(S)O-, -OC(S)NR f -、-NR f C(S)O-, -SS-, -S(O) 0-2 -, phenylene or pyridinylene, wherein the phenylene or pyridinylene is optionally substituted with one or more R*;

[0269] R* is independently H, halogen, cyano, C 1-10 Alkyl, C 1-10 Halogenated alkyl, -L f -OR f 、-L f -SR f or -L f -NR f R' f ;

[0270] L f independently a bond or C 1-8 alkylene;

[0271] R f and R' f Independently H, C 1-10 Alkyl, C 3-14 Cycloalkyl or 3-14 membered heterocyclic group;

[0272] R1 and R2 are independently C 4-20 Alkyl, C 4-20 Alkenyl or C 4-20 Alkynyl, which is optionally substituted by one or more R 1s substituted, and wherein one or more methylene units are optionally and independently replaced by -NR'-;

[0273] R 1s Independently H, C 1-20 Alkyl, -L c -OR c 、-L c -SR c or -L c -NR c R' c ;

[0274] R and R' are independently H or C 1-20 alkyl;

[0275] L c independently a bond or C 1-20 alkylene;

[0276] R c and R' c Independently H, C 1-20 Alkyl, C 3-14 Cycloalkyl or 3-14 membered heterocyclic group;

[0277] R3 is -CN, -OR g 、-C(O)R g 、-OC(O)R g 、-NR”C(O)R g 、-NR g R' g 、-NR”C(O)NR g R' g 、-NR”C(O)R g 、-NR”S(O)2R g 、-OC(O)NR g R' g 、-NR”C(O)OR g 、-N(OR g )C(O)R g 、-N(OR g )S(O)2R g 、-N(OR g )C(O)OR g 、-N(OR g )C(O)R g R' g, 3-14 membered heterocyclyl or 5-14 membered heteroaryl;

[0278] R g and R' g Independently H, C 1-10 Alkyl, C 3-10 Cycloalkyl and 3-10 membered heterocyclic groups;

[0279] R" is independently H or C 1-6 alkyl;

[0280] R5 and R6 are independently C 1-8 Alkyl, wherein the alkyl group is optionally substituted by one or more R 4s replace;

[0281] Or R5 and R6 together with the carbon atom to which they are attached form C 3-14 Cycloalkylene or 3-14 membered heterocyclylene, wherein the cycloalkylene and heterocyclylene are optionally substituted by one or more R 4s replace;

[0282] R 4s are independently H, halogen, cyano, C 1-8 Alkyl, C 1-8 Halogenated alkyl, -L d -OR d 、-L d -SR d or -L d -NR d R' d ;

[0283] L d independently a bond or C 1-8 alkylene;

[0284] R d and R' d Independently H, C 1-8 Alkyl, C 3-14 Cycloalkyl or 3-14 membered heterocyclic group.

[0285] In one embodiment, the ionizable lipid is a compound of formula (X):

[0286] or a stereoisomer, a mixture of stereoisomers or a pharmaceutically acceptable salt thereof, wherein

[0287] a is 1, 2, 3, 4, 5, or 6;

[0288] b is 4, 5, 6, 7, 8, 9, or 10;

[0289] c is 1, 2, 3, 4, 5, or 6;

[0290] d is 0, 1, 2, 3, or 4;

[0291] c+d is 3, 4, 5, 6, 7, 8 or 9.

[0292] In one embodiment, the ionizable lipid is:

[0293] or a stereoisomer, a mixture of stereoisomers or a pharmaceutically acceptable salt thereof.

[0294] In one embodiment, the ionizable lipid is:

[0295] or a stereoisomer, a mixture of stereoisomers or a pharmaceutically acceptable salt thereof.

[0296] In one embodiment, the ionizable lipid is DODAP, DODMA, DLinDMA, DLin-KC2-DMA, DLin-MC3-DMA, SM-102, or ALC-0315, or a combination thereof, or a stereoisomer, mixture of stereoisomers, or pharmaceutically acceptable salt thereof.

[0297] In one embodiment, the molar percentage of the ionizable lipids to the total lipids is 35-70 mol%, more preferably 40-60 mol%, such as 35 mol%, 40 mol%, 45 mol%, 50 mol%, 55 mol% or 60 mol%.

[0298] In one embodiment, the lipid nanoparticles further comprise a phospholipid. In one embodiment, the phospholipid is selected from any one or more of DSPC, DMPC, DOPC, DPPC, POPC, DOPE, DMPE, POPOE or DPPE.

[0299] In one embodiment, the molar percentage of the phospholipids to the total lipids is 5-30 mol%, preferably 5-20 mol%, more preferably 5-15 mol%, for example 5 mol%, 6 mol%, 7 mol%, 8 mol%, 9 mol%, 10 mol%, 11 mol%, 12 mol%, 13 mol%, 14 mol% or 15 mol%.

[0300] In one embodiment, the lipid nanoparticles further comprise a structural lipid. In one embodiment, the structural lipid is selected from the group consisting of cholesterol, campesterol, stigmasterol, sitosterol, brassicasterol, ergosterol, solanine, ursolic acid, α-tocopherol, β-sitosterol, avenasterol, and rapeseed oil sterol.

[0301] In one embodiment, the molar percentage of the structured lipids to the total lipids is 10-80 mol%, more preferably 15-70 mol%, more preferably 30-50 mol%, more preferably 30-40 mol%, for example 30 mol%, 35 mol%, 40 mol%, 45 mol%, 50 mol%, 55 mol% or 60 mol%.

[0302] In one embodiment, the lipid nanoparticle further comprises a polymer-conjugated lipid. In one embodiment, the polymer-conjugated lipid is a PEGylated lipid. In one embodiment, the PEGylated lipid comprises a PEGylated portion having a molecular weight of about 1000Da to about 20,000Da, about 1000Da to about 5000Da, or about 1000Da to about 2000Da. In one embodiment, the polymer-conjugated lipid is ALC-0159, DMG-PEG2000, DMPE-PEG1000, DPPE-PEG1000, DSPE-PEG1000, DOPE-PEG1000, ceramide-PEG2000, DMPE-PEG2000, DPPE-PEG2000, DSPE-PEG2000, DSPE-PEG2000-mannose, ceramide-PEG5000, DSPE-PEG5000, or DSPE-PEG2000-amino.

[0303] In one embodiment, the molar percentage of the polymer-conjugated lipid to the total lipid is 0.25-10 mol%, more preferably 0.5-5 mol%, more preferably 0.5-3 mol%, for example 0.5 mol%, 0.6 mol%, 0.7 mol%, 0.8 mol%, 0.9 mol%, 1.0 mol%, 1.1 mol%, 1.2 mol%, 1.3 mol%, 1.4 mol%, 1.5 mol%, 1.6 mol%, 1.7 mol%, 1.8 mol%. l%, 1.9mol%, 2.0mol%, 2.1mol%, 2.2mol%, 2.3mol%, 2.4mol%, 2.5mol%, 2.6mol%, 2.7mol%, 2.8mol%, 2.9mol %, 3.0 mol%, 3.1 mol%, 3.2 mol%, 3.3 mol%, 3.4 mol%, 3.5 mol%, 3.6 mol%, 3.7 mol%, 3.8 mol%, 3.9 mol%, 4.0 mol%.

[0304] In one embodiment, lipid nanoparticles include a payload, and wherein said payload is a therapeutic agent, a preventive agent or a diagnostic agent. In one embodiment, payload is a nucleic acid. In one embodiment, nucleic acid is an antisense oligonucleotide (ASO), DNA or RNA, optionally wherein said RNA is interference (RNAi), small interfering RNA (siRNA), short hairpin RNA (shRNA), antisense RNA (aRNA), messenger RNA (mRNA), modified messenger RNA (mmRNA), long non-coding RNA (lncRNA), microRNA (miRNA), small activation RNA (saRNA), polyencoded nucleic acid (MCNA), polymer-encoded nucleic acid (PCNA), guide RNA (gRNA), CRISPR RNA (crRNA) or any other RNA in ribozyme.

[0305] In one embodiment, the molar ratio of total N atoms in the steroid compound and ionizable lipid to P atoms in the nucleic acid molecule is from about 1:1 to about 20:1, from about 1:1 to about 20:1, from about 2:1 to about 10:1, or from about 4:1 to about 8:1.

[0306] In one embodiment, when lipid nanoparticles comprising a payload are systemically administered to a subject, the delivery or expression level of the payload is higher in the spleen of the subject than in the liver of the subject.

[0307] In one embodiment, the level of delivery or expression of the payload in the spleen of a subject is at least 1 fold, at least 2 fold, at least 3 fold, at least 4 fold, at least 5 fold, at least 10 fold, at least 20 fold, at least 30 fold, at least 40 fold, at least 50 fold, or at least 100 fold greater than the level of delivery or expression of the payload in the liver of a subject.

[0308] In one embodiment, provided herein are lipid nanoparticle compositions comprising the lipid nanoparticles described herein.

[0309] In one embodiment, provided herein is a pharmaceutical composition comprising a lipid nanoparticle or lipid nanoparticle composition described herein.

[0310] In one embodiment, provided herein is a method of treating or preventing a spleen disease or a spleen-related disease in a subject suffering from the spleen disease or the spleen-related disease, comprising administering a therapeutically effective amount of a lipid nanoparticle described herein, a lipid nanoparticle composition described herein, or a pharmaceutical composition described herein to the subject.

[0311] In one embodiment, provided herein is a use of a lipid nanoparticle as described herein or a lipid nanoparticle composition as described herein in the preparation of a medicament for preventing or treating a spleen disease or a spleen-related disease.

[0312] In one embodiment, provided herein is a lipid nanoparticle as described herein, a lipid nanoparticle composition as described herein, or a pharmaceutical composition as described herein for use in preventing or treating a spleen disease or a spleen-related disease.

[0313] In a preferred embodiment, the spleen disease or spleen-related disease mentioned above includes a blood system disease, an infectious disease, an immune disease or cancer.

[0314] In a preferred embodiment, the spleen diseases or spleen-related diseases mentioned above include hemolytic anemia, thrombocytopenic purpura, chronic leukemia, lymphoma, such as Hodgkin's lymphoma or non-Hodgkin's lymphoma, myelodysplastic syndrome, myelofibrosis, splenomegaly, lipid metabolism disorders, such as glucocerebrosidosis and sphingomyelinosis, splenomegaly and hypersplenism associated with sepsis, typhoid fever, infectious mononucleosis or subacute bacterial endocarditis, splenomegaly and hypersplenism associated with malaria, tuberculosis, kala-azar, etc., congestive splenomegaly and hypersplenism caused by portal hypertension in cirrhosis, splenic cysts, such as parasitic splenic cysts or pseudosplenic cysts, splenic abscesses, primary splenic tumors, splenic metastases, splenic artery aneurysms, splenic infarction, splenic purpura, splenic rupture, etc.

[0315] In one embodiment, provided herein is a method for preparing a lipid nanoparticle described herein or a lipid nanoparticle composition described herein, comprising the steps of:

[0316] (i) dissolving a mixture comprising lipids in a first solution, wherein the lipid solution is formed in an organic solvent;

[0317] (ii) dissolving the payload in the second solution to form a payload solution; and

[0318] (iii) The lipid solution and the payload solution are mixed to form lipid nanoparticles.

[0319] In one embodiment, mixing is performed using a microfluidic system.

[0320] In another aspect, provided herein is a steroid compound of formula (II-A):

[0321] or a stereoisomer, a mixture of stereoisomers, or a pharmaceutically acceptable salt thereof, wherein:

[0322] Each are independently a single bond or a double bond;

[0323] Y is -C(O)O-, -O-, -NH-, -SC(O)O-, -OC(O)NR 2a -、-NR 2a C(O)NR 2a -、-OC(O)S-、-OC(O)O-、-NR 2a C(O)O-, -OC(O)-, -SC(O)-, -C(O)S-, -NR 2a -、-C(O)NR 2a -、-NR 2a C(O)-、-NR 2a C(O)S-、-SC(O)NR 2a -, -C(O)-, -OC(S)-, -C(S)O-, -OC(S)NR 2a -、-NR 2a C(S)O- or -S(O) 0-2 -, and wherein the left side is connected in the direction of V1;

[0324] Y1 is absent or selected from -C(O)O-, -O-, -NH-, -SC(O)O-, -OC(O)NR 2a -、-NR 2a C(O)NR 2a -、-OC(O)S-、-OC(O)O-、-NR 2aC(O)O-, -OC(O)-, -SC(O)-, -C(O)S-, -NR 2a -、-C(O)NR 2a -、-NR 2a C(O)-、-NR 2a C(O)S-、-SC(O)NR 2a -, -C(O)-, -OC(S)-, -C(S)O-, -OC(S)NR 2a -、-NR 2a C(S)O- or -S(O) 0-2 -, and wherein the left side is connected in the direction of V1;

[0325] R 2a Each instance of is independently H, C 1-10 Alkyl, C 3-14 cycloalkyl or 3 to 14 membered heterocyclyl, and wherein the alkyl, cycloalkyl and heterocyclyl are independently optionally replaced by C 1-10 Alkyl, L 2b -OR 2b , Lb-SR 2b 、L 2b -NR 2b R' 2b 、L 2b N + R 2b R' 2b R” 2b 、L 2b -OC(O)R 2b or L 2b -C(O)OR 2b replace;

[0326] R 201 、R 202 、R 203 、R 204 Each example is independently H, OH, alkyl, alkoxy, -L 2b -NR 21 R 22 or -L 2b -N + R 21 R 22 R 23 、L 2b -OC(O)R 21 or L 2b -C(O)OR 21 ;

[0327] M is an integer from 3 to 12;

[0328] m1 is an integer from 0 to 12;

[0329] V1 is NR 21 、N + R 21 R 22 R 23 , imidazole, pyridine, pyrrole, pyrrolidine, pyrazole, (iso)thiazole, (iso)oxazole, oxadiazine, oxazoline, dithioureaazole, (iso)triazole, tetrazole, pentazole, indole, indoline, pyrimidine, pyrazine, quinazoline, piperazine, piperidine, morpholine, pyran, quinoxaline, quinoline, quinolinone, (iso)quinoline, amidine, guanidine or urea;

[0330] M is absent or is one or more anions independently selected from chloride, bromide, iodide, sulfate, nitrate, perchlorate, formate, citrate, sulfonate, methanesulfonate, trifluoromethanesulfonate, trifluoroacetate, pyrosulfate, bisulfate, sulfite, bisulfite, phosphate, pyrophosphate, monohydrogen phosphate, dihydrogen phosphate, acetate, propionate, decanoate, octanoate, acrylate, isobutyrate, hexanoate, heptanoate, oxalate, malonate, succinate, fumarate, maleate, benzoate, phthalate, phenylacetate, lactate, glycolate, tartrate, mandelate, methanesulfonate, gluconate, aspartate, or glutamate;

[0331] R 21 、R 22 、R 23 are each independently H, optionally substituted C 1-8 Alkyl, -L 2b -OR 2b 、-L 2b -SR 2b 、-L 2b -NR 2b R' 2b R” 2b 、L 2b -OC(O)R 2b or L 2b -C(O)OR 2b ;

[0332] L 2b is absent or optionally substituted C 1-10 alkylene;

[0333] R 2b , R' 2b and R” 2b Each of is independently H or C 1-10 alkyl; and

[0334] R z1 It is C 1-14 Alkyl or C1-14 alkenyl, wherein the alkyl and alkenyl groups are optionally substituted; provided that it is not:

[0335] In one embodiment, the present invention provides a steroid compound having the formula (II-B):

[0336] or a stereoisomer, a mixture of stereoisomers, or a pharmaceutically acceptable salt thereof, wherein:

[0337] Y is -C(O)O-, -O-, -NH-, -SC(O)O-, -OC(O)NR 2a -、-NR 2a C(O)NR 2a -、-OC(O)S-、-OC(O)O-、-NR 2a C(O)O-, -OC(O)-, -SC(O)-, -C(O)S-, -NR 2a -、-C(O)NR 2a -、-NR 2a C(O)-、-NR 2a C(O)S-、-SC(O)NR 2a -, -C(O)-, -OC(S)-, -C(S)O-, -OC(S)NR 2a -、-NR 2a C(S)O- or -S(O) 0-2 -, and wherein the leftward connection is in the direction of V1;

[0338] Y1 is absent, -C(O)O-, -O-, -NH-, -SC(O)O-, -OC(O)NR 2a -、-NR 2a C(O)NR 2a -、-OC(O)S-、-OC(O)O-、-NR 2a C(O)O-, -OC(O)-, -SC(O)-, -C(O)S-, -NR 2a -、-C(O)NR 2a -、-NR 2a C(O)-、-NR 2a C(O)S-、-SC(O)NR 2a -, -C(O)-, -OC(S)-, -C(S)O-, -OC(S)NR 2a -、-NR 2a C(S)O- or -S(O) 0-2 -, and wherein the leftward connection is in the direction of V1;

[0339] R 2a Each instance of is independently H, C 1-10 Alkyl, C 3-14 cycloalkyl or 3 to 14 membered heterocyclyl, and wherein the alkyl, cycloalkyl and heterocyclyl are independently optionally replaced by C 1-10 Alkyl, L 2b -OR 2b 、L b -SR 2b 、L 2b -NR 2b R' 2b 、L 2b N + R 2b R' 2b R” 2b 、L 2b -OC(O)R 2b or L 2b -C(O)OR 2b replace;

[0340] R 201 、R 202、 R 203 、R 204 Each example is independently H, OH, alkyl, alkoxy, -L 2b -NR 21 R 22 or -L 2b -N + R 21 R 22 R 23 、L 2b -OC(O)R 21 or L 2b -C(O)OR 21 ;

[0341] m is an integer from 5 to 12;

[0342] m1 is an integer from 0 to 12;

[0343] V1 is NR 21 R 22 、N + R 21 R 22 R 23 , imidazole, pyridine, pyrrole, pyrrolidine, pyrazole, (iso)thiazole, (iso)oxazole, oxadiazine, oxazoline, dithioureaazole, (iso)triazole, tetrazole, pentazole, indole, indoline, pyrimidine, pyrazine, quinazoline, piperazine, piperidine, morpholine, pyran, quinoxaline, quinoline, quinolinone, (iso)quinoline, amidine, guanidine or urea;

[0344] M is absent or is one or more anions independently selected from chloride, bromide, iodide, sulfate, nitrate, perchlorate, formate, citrate, sulfonate, methanesulfonate, trifluoromethanesulfonate, trifluoroacetate, pyrosulfate, bisulfate, sulfite, bisulfite, phosphate, pyrophosphate, monohydrogen phosphate, dihydrogen phosphate, acetate, propionate, decanoate, octanoate, acrylate, isobutyrate, hexanoate, heptanoate, oxalate, malonate, succinate, fumarate, maleate, benzoate, phthalate, phenylacetate, lactate, glycolate, tartrate, mandelate, methanesulfonate, gluconate, aspartate, or glutamate;

[0345] R 21 、R 22 、R 23 are each independently H, optionally substituted C 1-8 Alkyl, -L 2b -OR 2b 、-L 2b -SR 2b 、-L 2b -NR 2b R' 2b R” 2b 、L 2b -OC(O)R 2b or L 2b -C(O)OR 2b ;

[0346] L 2b is absent or optionally substituted C 1-10 alkylene;

[0347] R 2b , R' 2b and R” 2b Each of is independently H or C 1-10 alkyl.

[0348] In one embodiment, the present invention provides a steroid compound of formula (IP):

[0349] or a stereoisomer, a mixture of stereoisomers, or a pharmaceutically acceptable salt thereof, wherein:

[0350] Is a single bond or a double bond;

[0351] Y1 does not exist, -C(O)O-, -O-, -NH-, -SC(O)O-, -OC(O)NR 2a -、-NR 2a C(O)NR 2a-、-OC(O)S-、-OC(O)O-、-NR 2a C(O)O-, -OC(O)-, -SC(O)-, -C(O)S-,-NR 2a -,-C(O)NR 2a -,-NR 2a C(O)-,-NR 2a C(O)S-,-SC(O)NR 2a -,-C(O)-,-OC(S)-, -C(S)O-, -OC(S)NR 2a -、-NR 2a C(S)O-, -S(O) 0-2 -or -OPO3-, and wherein the leftward connection is in the direction of V1;

[0352] R 2a Each instance of is independently H, C 1-10 Alkyl, C 3-14 cycloalkyl or 3 to 14 membered heterocyclyl, and wherein the alkyl, cycloalkyl and heterocyclyl are independently optionally replaced by C 1-10 Alkyl, L 2b -OR 2b 、L b -SR 2b 、L 2b -NR 2b R' 2b 、L 2b N + R 2b R' 2b R” 2b 、L 2b -OC(O)R 2b or L 2b -C(O)OR 2b replace;

[0353] R 201 、R 202、 R 203 、R 204 Each example is independently H, OH, alkyl, alkoxy, -L 2b -NR 21 R 22 、-L 2b -N + R 21 R 22 R 23 、L 2b -OC(O)R 21 or L 2b -C(O)OR 21 ;

[0354] R205 Absence, cations, H, C 1-6 Alkyl, C 1-6 Halogenated alkyl, C 2-6 Alkenyl or C 2-6 alkynyl, and wherein alkyl, haloalkyl, alkenyl and alkynyl are optionally substituted;

[0355] m is an integer from 1 to 12;

[0356] m1 is an integer from 0 to 12;

[0357] V1 is NR 21 R 22 、N + R 21 R 22 R 23 , imidazole, pyridine, pyrrole, pyrrolidine, pyrazole, (iso)thiazole, (iso)oxazole, oxadiazine, oxazoline, dithioureaazole, (iso)triazole, tetrazole, pentazole, indole, indoline, pyrimidine, pyrazine, quinazoline, piperazine, piperidine, morpholine, pyran, quinoxaline, quinoline, quinolinone, (iso)quinoline, amidine, guanidine or urea;

[0358] M is absent or is one or more anions independently selected from chloride, bromide, iodide, sulfate, nitrate, perchlorate, formate, citrate, sulfonate, methanesulfonate, trifluoromethanesulfonate, trifluoroacetate, pyrosulfate, bisulfate, sulfite, bisulfite, phosphate, pyrophosphate, monohydrogen phosphate, dihydrogen phosphate, acetate, propionate, decanoate, octanoate, acrylate, isobutyrate, hexanoate, heptanoate, oxalate, malonate, succinate, fumarate, maleate, benzoate, phthalate, phenylacetate, lactate, glycolate, tartrate, mandelate, methanesulfonate, gluconate, aspartate, or glutamate;

[0359] R 21 、R 22 、R 23 Each instance of is independently H, optionally substituted C 1-8 Alkyl, -L 2b -OR 2b 、-L 2b -SR 2b 、-L 2b -NR 2b R' 2b R” 2b 、L 2b -OC(O)R 2b or L 2b -C(O)OR 2b ;

[0360] L 2b is absent or optionally substituted C 1-10 alkylene;

[0361] R 2b , R' 2b and R” 2b Each instance of is independently H or C 1-10 alkyl;

[0362] R z1 It is C 1-14 Alkyl or C 1-14 alkenyl, wherein the alkyl and alkenyl groups are optionally substituted.

[0363] In one embodiment, provided herein is a steroid compound of formula (II-C1):

[0364] or a stereoisomer, a mixture of stereoisomers, or a pharmaceutically acceptable salt thereof, wherein:

[0365] R z1 yes

[0366] R 2a is H, optionally substituted C 1-8 Alkyl, -L 2b -OR 2b 、L 2b -OC(O)R 2b or L 2b -C(O)OR 2b , preferably H, -CH3, -CH2C(O)OCH3 or -CH2C(O)OH;

[0367] V1 is NR 21 R 22 or N + R 21 R 22 R 23 ; where R 21 、R 22 、R 23 are each independently H, optionally substituted C 1-8 Alkyl, -L 2b -OR 2b 、-L 2b -SR 2b 、-L 2b -NR 2b R' 2b R” 2b 、L 2b -OC(O)R 2b or L2b -C(O)OR 2b ; and R 21 and R 22 Not simultaneously hydrogen;

[0368] L 2b is absent or optionally substituted C 1-10 alkylene;

[0369] R 2b , R' 2b and R” 2b Each of is independently H or C 1-10 alkyl;

[0370] M is absent or is one or more anions independently selected from chloride, bromide, iodide, sulfate, nitrate, perchlorate, formate, citrate, sulfonate, methanesulfonate, trifluoromethanesulfonate, trifluoroacetate, pyrosulfate, bisulfate, sulfite, bisulfite, phosphate, pyrophosphate, monohydrogen phosphate, dihydrogen phosphate, acetate, propionate, decanoate, octanoate, acrylate, isobutyrate, hexanoate, heptanoate, oxalate, malonate, succinate, fumarate, maleate, benzoate, phthalate, phenylacetate, lactate, glycolate, tartrate, mandelate, methanesulfonate, gluconate, aspartate, or glutamate;

[0371] m is an integer from 1 to 10, for example, 1, 2, 3, 4, 5, 6, 7, 8, 9 or 10.

[0372] In one embodiment, provided herein is a steroid compound of formula (II-C2):

[0373] or a stereoisomer, a mixture of stereoisomers, or a pharmaceutically acceptable salt thereof, wherein:

[0374] R z1 yes

[0375] V1 is NR 21 R 22 or N + R 21 R 22 R 23 ; where R 21 、R 22 、R 23 are each independently H, optionally substituted C 1-8 Alkyl, -L 2b -OR 2b 、-L 2b -SR2b 、-L 2b -NR 2b R' 2b R” 2b 、L 2b -OC(O)R 2b or L 2b -C(O)OR 2b ; and R 21 and R 22 Not simultaneously hydrogen;

[0376] L 2b is absent or optionally substituted C 1-10 alkylene;

[0377] R 2b , R' 2b and R” 2b Each of is independently H or C 1-10 alkyl;

[0378] M is absent or is one or more anions independently selected from chloride, bromide, iodide, sulfate, nitrate, perchlorate, formate, citrate, sulfonate, methanesulfonate, trifluoromethanesulfonate, trifluoroacetate, pyrosulfate, bisulfate, sulfite, bisulfite, phosphate, pyrophosphate, monohydrogen phosphate, dihydrogen phosphate, acetate, propionate, decanoate, octanoate, acrylate, isobutyrate, hexanoate, heptanoate, oxalate, malonate, succinate, fumarate, maleate, benzoate, phthalate, phenylacetate, lactate, glycolate, tartrate, mandelate, methanesulfonate, gluconate, aspartate, or glutamate;

[0379] m is an integer from 1 to 10, for example, 1, 2, 3, 4, 5, 6, 7, 8, 9 or 10.

[0380] In one embodiment, the steroid compound is selected from:

[0381] or a stereoisomer, a mixture of stereoisomers or a pharmaceutically acceptable salt thereof.

[0382] In one embodiment, the steroid compound is:

[0383] or a stereoisomer, a mixture of stereoisomers or a pharmaceutically acceptable salt thereof.

[0384] In another aspect, provided herein is a steroid compound selected from:

[0385] or a stereoisomer, a mixture of stereoisomers or a pharmaceutically acceptable salt thereof.

[0386] In one embodiment, provided herein is a lipid nanoparticle comprising a steroid compound described herein. 4. Description of the Figures

[0387] FIG1 shows the expression levels in the liver, lung, and spleen of mice after administration of LNP5 in Experimental Example 3. The three rows in the figure represent the experimental results of three mice respectively. 5. Specific implementation methods

[0388] Provided herein are lipid nanoparticle compositions and their preparation and use.

[0389] In one aspect, a lipid nanoparticle (LNP, such as the LNP described in Chapter 0) is provided herein. In one embodiment, the LNP comprises a steroid compound with a cationic group, also referred to as a steroid ionizable lipid (cationic steroid compound, Chapter 0). In one embodiment, the LNP comprises an ionizable lipid (Chapter 0). In one embodiment, the LNP further comprises a phospholipid (Chapter 0). In one embodiment, the LNP does not comprise a phospholipid. In one embodiment, the LNP further comprises a structural lipid (Chapter 0). In one embodiment, the LNP further comprises a polymer-conjugated lipid, such as a pegylated lipid (Chapter 0). In one embodiment, the LNP comprises a payload (Chapter 0).

[0390] In one aspect, provided herein is a lipid nanoparticle composition (Section 0).

[0391] In one embodiment, the LNPs described herein preferentially deliver their payload to non-hepatic organs (eg, spleen) when administered to a subject. In one embodiment, provided herein are LNP compositions for use in treating spleen diseases or spleen-related diseases.

[0392] In another aspect, provided herein is a pharmaceutical composition comprising the LNP provided herein (Section 0).

[0393] Also provided herein is a method for preparing the LNP compositions described herein (see Section 0). Also provided herein is a method for treating a disease using the LNP compositions described herein (Section 0). In one embodiment, provided herein is a method for treating a spleen disease or spleen-related disease.

[0394] One aspect of the application relates to the discovery that including a steroid compound (e.g., a cationic steroid compound as provided in Section 0) in LNP significantly increases the efficiency of delivery of the LNP and its payload to the spleen. More specifically, cationic steroid compounds make the delivery of the LNP and its payload to the lungs more efficient than other cationic lipids (e.g., DOTAP). Cationic steroid compounds can be effective in relatively low amounts, which can reduce toxicity and side effects.

[0395] 5.1 Definition

[0396] 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. All patents, applications, published applications, and other publications are incorporated by reference in their entirety. If a term is defined in multiple ways herein, the definition in this section prevails unless otherwise stated.

[0397] The term "about" is used herein to mean approximately, roughly, roughly, or generally. When the term "about" is used in conjunction with a numerical range, it modifies that range by expanding the boundaries above and below the numerical values ​​recited. Generally, unless otherwise stated or otherwise apparent from the context, the term "about" can modify a numerical value above and below the recited value by a variance of 10%, up or down (higher or lower).

[0398] It should be noted that if there is a discrepancy between a described structure and the name of that structure, the described structure will be given greater weight.

[0399] When a range of values ​​is listed, it is intended to encompass every value and sub-range within the stated range. For example, "C 1-6 "Alkyl" is intended to include C1, C2, C3, C4, C5, C6, C 1-6 、C 1-5 、C 1-4 、C 1-3 、C 1-2 、C 2-6 、C 2-5 、C 2-4 、C 2-3 、C 3-6 、C3-5、C 3-4 、C 4-6 、C 4-5 and C 5-6 alkyl.

[0400] As used herein and unless otherwise indicated, "alkyl" refers to a straight-chain or branched saturated hydrocarbon group formed by removing one hydrogen atom. For example, "C 1-24"Alkyl" refers to a straight chain or branched hydrocarbon group having 1 to 24 carbon atoms. 1-20 "Alkyl" refers to a straight-chain or branched saturated hydrocarbon group having 1 to 20 carbon atoms. 6-24 "Alkyl" refers to a straight or branched saturated hydrocarbon group having 6 to 24 carbon atoms. 6-20 "Alkyl" refers to a free radical of a straight or branched saturated hydrocarbon group having 6 to 20 carbon atoms. In some embodiments, the alkyl group is C 1-24 Alkyl, C 4-20 Alkyl, C 6-20 Alkyl, C 8-20 Alkyl, C 10-20 Alkyl, C 6-18 Alkyl, C 6-16 Alkyl, C 6-14 Alkyl, C 6-12 Alkyl, C 8-10 Alkyl, C 2-8 Alkyl, C 7-9 Alkyl, C 4-6 Alkyl, C 1-16 Alkyl, C 1-14 Alkyl, C 2-14 Alkyl, C 1-13 Alkyl, C 1-12 Alkyl, C 1-10 Alkyl, C 1-8 Alkyl, C 1-7 Alkyl, C 2-7 Alkyl, C 1-6 Alkyl, C 1-5 Alkyl, C5 alkyl, C 1-4 Alkyl, C 2-4 Alkyl, C 1-3 Alkyl, C 2-3 Alkyl, C 1-2 Alkyl or methyl. C 1-6Examples of alkyl groups include methyl (C1), ethyl (C2), n-propyl (C3), isopropyl (C3), n-butyl (C4), tert-butyl (C4), sec-butyl (C4), isobutyl (C4), n-pentyl (C5), 3-pentyl (C5), pentyl (C5), neopentyl (C5), 3-methyl-2-butyl (C5), tert-pentyl (C5) and n-hexyl (C6). The term "alkyl" also includes heteroalkyl groups in which one or more (e.g., 1, 2, 3 or 4) carbon atoms are replaced by heteroatoms (e.g., oxygen, sulfur, nitrogen, boron, silicon, phosphorus). Alkyl groups may be substituted or unsubstituted. Alkyl groups may optionally be substituted by one or more substituents, for example, by 1 to 5 substituents, 1 to 3 substituents or 1 substituent. Common abbreviations for alkyl groups include Me(-CH3), Et(-CH2CH3), iPr(-CH(CH3)2), nPr(-CH2CH2CH3), n-Bu(-CH2CH2CH2CH3), or i-Bu(-CH2CH(CH3)2).

[0401] As used herein and unless otherwise specified, "alkenyl" refers to a straight-chain or branched unsaturated hydrocarbon group having at least one carbon-carbon double bond. For example, "C 6-24 "Alkenyl" refers to a straight or branched hydrocarbon group having 6 to 24 carbon atoms and at least one carbon-carbon double bond. 4-28 "Alkenyl" refers to a straight or branched hydrocarbon group having 4 to 28 carbon atoms and at least one carbon-carbon double bond. 4-20 "Alkenyl" refers to a straight or branched hydrocarbon group having 4 to 20 carbon atoms and at least one carbon-carbon double bond. "C2-14 alkenyl" refers to a straight or branched hydrocarbon group having 2 to 14 carbon atoms and at least one carbon-carbon double bond. In some embodiments, alkenyl is C 4-24 Alkenyl, C 6-24 Alkenyl, C 8-24 Alkenyl, C 10-24 Alkenyl, C8 alkenyl, C9 alkenyl, C 10 Alkenyl, C 11 Alkenyl, C 12 Alkenyl, C 13 Alkenyl, C 14 Alkenyl, C 15 Alkenyl, C 16 Alkenyl, C 2-13 Alkenyl, C 2-10 Alkenyl, C 2-6 Alkenyl or C 2-4 Alkenyl. C 2-6The example of alkenyl includes vinyl (C2), 1-propenyl (C3), 2-propenyl (C3), 1-butenyl (C4), 2-butenyl (C4), butadienyl (C4), pentenyl (C5), pentadienyl (C5), hexenyl (C6) etc.Term " alkenyl " also includes heteroalkenyl, wherein one or more (for example, 1,2,3 or 4) carbon atoms are replaced by heteroatoms (for example, oxygen, sulfur, nitrogen, boron, silicon, phosphorus).Alkenyl can be substituted or unsubstituted.Alkenyl can be optionally substituted by one or more substituents, for example, substituted by 1 to 5 substituents, 1 to 3 substituents or 1 substituent.

[0402] As used herein, and unless otherwise indicated, "alkynyl" refers to a straight or branched hydrocarbon group having at least one carbon-carbon triple bond. For example, "C 4-20 "Alkynyl" refers to a straight or branched hydrocarbon group having 2 to 20 carbon atoms and at least one carbon-carbon triple bond. In some embodiments, the alkynyl group is C 4-24 Alkynyl, C 6-20 Alkynyl, C 8-18 Alkynyl, C8 alkynyl, C9 alkynyl, C 10 Alkynyl, C 11 Alkynyl, C 12 Alkynyl, C 13 Alkynyl, C 14 Alkynyl, C 15 Alkynyl, C 16 Alkynyl, C 17 Alkynyl, C 2-13 Alkynyl, C 2-10 Alkynyl, C 2-6 Alkynyl or C 2-4 Alkynyl. C 2-6 Examples of alkynyl include, but are not limited to, ethynyl (C2), 1-propynyl (C3), 2-propynyl (C3), 1-butynyl (C4), 2-butynyl (C4), pentynyl (C5), hexynyl (C6) and the like. The term "alkynyl" also includes heteroalkynyls, wherein one or more (e.g., 1, 2, 3 or 4) carbon atoms are replaced by heteroatoms (e.g., oxygen, sulfur, nitrogen, boron, silicon, phosphorus). Alkynyl groups can be substituted or unsubstituted. Alkynyl groups can be selected to be substituted by one or more substituents, for example, by 1 to 5 substituents, 1 to 3 substituents or 1 substituent.

[0403] As used herein and unless otherwise indicated, "alkylene" refers to a divalent group formed by removing another hydrogen atom from an alkyl group. For example, "C 1-24 "Alkylene" refers to a 1-24 A divalent group formed by removing another hydrogen from an alkyl group. In some embodiments, an alkylene group is a C 1-24 Alkylene, C 4-24Alkylene, C 1-20 Alkylene, C 8-20 Alkylene, C8 alkylene, C9 alkylene, C 10 Alkylene, C 11 Alkylene, C 12 Alkylene, C 13 Alkylene, C 13 Alkylene, C 14 Alkylene, C 15 Alkylene, C 16 Alkylene, C 17 Alkylene, C 18 Alkylene, C 1-20 Alkylene, C 1-14 Alkylene, C 2-14 Alkylene, C 1-13 Alkylene, C 1-12 Alkylene, C 1-10 Alkylene, C 1-8 Alkylene, C 7-9 Alkylene, C 4-6 Alkylene, C 1-7 Alkylene, C 2-7 Alkylene, C 1-6 Alkylene, C 1-5 Alkylene, C5 alkylene, C 1-4 Alkylene, C 2-4 Alkylene, C 1-3 Alkylene, C 2-3 Alkylene, C 1-2 Alkylene or methylene. Alkylene can be substituted or unsubstituted. Unsubstituted alkylene groups include, but are not limited to, methylene (-CH2-), ethylene (-CH2CH2-), propylene (-CH2CH2CH2-), butylene (-CH2CH2CH2CH2-), pentylene (-CH2CH2CH2CH2CH2-), hexylene (-CH2CH2CH2CH2CH2CH2-), and the like. Examples of substituted alkylene groups (e.g., alkylene groups substituted with one or more alkyl(methyl) groups) include, but are not limited to, substituted methylene (-CH(CH3)-, -C(CH3)2-), substituted ethylene (-CH(CH3)CH2-, -CH2CH(CH3)-, -C(CH3)2CH2-, -CH2C(CH3)2-), substituted propylene (-CH(CH3)CH2CH2-, -CH2CH(CH3)CH2-, -CH2CH2CH(CH3)-, -C(CH3)2CH2CH2-, -CH2C(CH3)2CH2-), and the like.

[0404] As used herein and unless otherwise indicated, "alkenylene" refers to a divalent group formed by removing one hydrogen from an alkenyl group. For example, "C 2-24 "Alkenylene" refers to C 2-24 In some embodiments, alkenyl is a C 4-24 Alkenylene, C 4-20 Alkenylene, C 8-20 Alkenylene, C 8-18 Alkenylene, C 8-16 Alkenylene, C 4-14 Alkenylene, C 2-13 Alkenylene, C 2-10 Alkenylene, C 2-6 Alkenylene or C 2-4 Alkenylene. Alkenylene can be substituted or unsubstituted. Exemplary unsubstituted alkenylene groups include, but are not limited to, ethenylene (-CH=CH-) and propenylene (e.g., -CH=CHCH2-, -CH2-CH=CH-). Exemplary substituted alkenylene groups (e.g., alkenylene groups substituted with one or more alkyl (methyl) groups) include, but are not limited to, substituted ethenylene (-C(CH3)=CH-, -CH=C(CH3)-), substituted propenylene (e.g., -C(CH3)=CHCH2-, -CH=C(CH3)CH2-, -CH=CHCH(CH3)-, -CH=CHC(CH3)2-, -CH(CH3)-CH=CH-, -C(CH3)2-CH=CH-, -CH2-C(CH3)=CH-, -CH2-CH=C(CH3)-), and the like.

[0405] As used herein and unless otherwise indicated, "alkynylene" refers to a divalent group formed by removing one hydrogen from an alkynyl group. For example, "C 2-24 "Alkynylidene" refers to C 2-24 An alkynyl group in which another hydrogen is removed to provide a divalent radical of an alkynylene group. "C 2-13 "Alkynylene" refers to a C2-13 alkynyl group wherein another hydrogen is removed to provide a divalent radical of an alkynylene group. "C 4-14 "Alkynylidene" refers to C 4-14 An alkynyl group wherein another hydrogen is removed to provide a divalent radical of an alkynylene group. In some embodiments, an alkynylene group is C 2-24 Alkynylidene, C 2-20 Alkynylidene, C 4-16 Alkynylidene, C 8-16 Alkynylidene, C 9-16 Alkynylidene, C 10-16 Alkynylidene, C 4-12 Alkynylidene, C2-10 Alkynylidene, C 2-6 Alkynylidene or C 2-4 Alkynylidene. Alkynylidene can be substituted or unsubstituted. Exemplary alkynylene groups include, but are not limited to, ethynylene (-C≡C-), substituted or unsubstituted propynylene (-C≡CCH2-), and the like.

[0406] “C 0-6 "Alkylene" refers to the chemical bond described above and "C 1-6 Alkylene", "C 0-4 "Alkylene" refers to the chemical bond described above and "C 1-4 "Alkylene".

[0407] The term "the total length of variable A and variable B is x carbon atoms" means that the total number of carbon atoms in the main chain of the group represented by variable A and the number of carbon atoms in the main chain of the group represented by variable B is x.

[0408] "Halo" or "halogen" refers to fluorine (F), chlorine (Cl), bromine (Br), or iodine (I).

[0409] "Haloalkyl" refers to an alkyl group substituted with one or more halogens. For example, "C 1-10 "Haloalkyl" refers to the above-mentioned "C 1-10 In some embodiments, the haloalkyl group is C 1-24 Halogenated alkyl, C 1-20 Halogenated alkyl, C 1-18 Halogenated alkyl, C 1-16 Halogenated alkyl, C 1-14 Halogenated alkyl, C 1-12 Halogenated alkyl, C 6-12 Halogenated alkyl, C 1-6 Halogenated alkyl, C 1-4 Haloalkyl or C 1-2 In some embodiments, haloalkyl includes but is not limited to C haloalkyl, C haloalkyl, C haloalkyl, C haloalkyl, C haloalkyl, C haloalkyl or C haloalkyl.Haloalkyl can optionally further comprise one or more non-halogen replacements.Exemplary haloalkyl includes but is not limited to-CF ,-CH F ,-CHF ,-CHFCH F ,-CH CHF ,-CF CF ,-CCl ,-CH Cl ,-CHCl , 2,2,2-trifluoro-1,1-dimethyl-ethyl etc.Haloalkyl can be substituted at any available point of attachment, for example, replaced by 1 to 5 substituents, 1 to 3 substituents or 1 substituent.

[0410] As used herein and unless otherwise specified, "cycloalkyl" refers to a non-aromatic monocyclic or polycyclic hydrocarbon group consisting solely of carbon and hydrogen atoms. For example, "C 3-14 "Cycloalkyl" or "3- to 14-membered cycloalkyl" refers to a non-aromatic cyclic hydrocarbon radical having 3 to 14 ring carbon atoms and zero heteroatoms, optionally containing 1, 2, or 3 double or triple bonds. In some embodiments, the cycloalkyl radical is a 3- to 10-membered cycloalkyl radical (C 3-10 cycloalkyl), 5- to 10-membered cycloalkyl (C 5-10 cycloalkyl), 3 to 8 membered cycloalkyl (C 3-8 cycloalkyl), 3 to 7 membered cycloalkyl (C 3-7 cycloalkyl), 3 to 6 membered cycloalkyl (C 3-6 cycloalkyl), 5 to 7 membered cycloalkyl (C 5-7 cycloalkyl), 4 to 6 membered cycloalkyl (C 4-6 cycloalkyl), 5- to 6-membered cycloalkyl (C 5-6 Cycloalkyl groups include cycloalkyl groups (Ccycloalkyl), ...). Cycloalkyl groups also include ring systems in which the cycloalkyl rings described above are fused to one or more aryl or heteroaryl groups, wherein the point of attachment is on the cycloalkyl ring, and in such cases, the number of carbon atoms continues to refer to the number of carbon atoms in the cycloalkyl system. Cycloalkyl groups further include cycloalkyl rings described above in which substituents on any non-adjacent carbon atoms are linked to form a bridged ring, thereby together forming a polycycloalkane sharing two or more carbon atoms. Cycloalkyl groups further include cycloalkyl rings described above in which substituents on the same carbon atom are linked to form a ring, thereby together forming a polycycloalkane sharing one carbon atom. Exemplary cycloalkyl groups include, but are not limited to, cyclopropyl (C3), cyclopropenyl (C3), cyclobutyl (C4), cyclobutenyl (C4), cyclopentyl (C5), cyclopentenyl (C5), cyclohexyl (C6), cyclohexenyl (C6), cyclohexadienyl (C6), cycloheptyl (C7), cycloheptenyl (C7), cycloheptadienyl (C7), cycloheptatrienyl (C7), etc. Cycloalkyl can be substituted or unsubstituted. Cycloalkyl can be substituted by one or more substituents, for example, substituted by 1 to 5 substituents, 1 to 3 substituents, or 1 substituent.

[0411] As used herein and unless otherwise indicated, "cycloalkylene" refers to a divalent radical formed by removing another hydrogen from a cycloalkyl group. For example, "C 3-14 "Cycloalkylene" refers to a cycloalkylene group formed by removing a C 3-10 In some embodiments, cycloalkylene is C 3-12Cycloalkylene, C 3-8 Cycloalkylene, C 3-6 Cycloalkylene, C 3-5 Cycloalkylene, C 3-4 Cycloalkylene, C3 cycloalkylene, C4 cycloalkylene, C5 cycloalkylene or C6 cycloalkylene.Cycloalkylene can be substituted or unsubstituted.Exemplary cycloalkylene include cyclopropylene, cyclobutylene, cyclopentylene or cyclohexylene.

[0412] As used herein and unless otherwise indicated, "heterocyclyl" refers to a saturated or unsaturated radical of a non-aromatic ring system having ring carbon atoms and at least one ring heteroatom. For example, a "3 to 14 membered heterocyclyl" refers to a saturated or unsaturated radical of a 3 to 14 membered non-aromatic ring system having at least one ring heteroatom (e.g., 1-5 heteroatoms), optionally containing 1, 2 or 3 double or triple bonds. In some embodiments, the ring heteroatoms in the heterocyclyl are selected from one or more of nitrogen, oxygen, sulfur, boron, phosphorus or silicon. In heterocyclyls containing one or more nitrogen atoms, the point of attachment to the rest of the molecule may be a carbon atom or a nitrogen atom, as long as valence permits. In some embodiments, the heterocyclyl group is a 3- to 10-membered heterocyclyl, a 5- to 10-membered heterocyclyl, a 5- to 10-membered nitrogen-containing heterocyclyl, a 3- to 8-membered heterocyclyl, a 3- to 8-membered nitrogen-containing heterocyclyl, a 3- to 7-membered heterocyclyl, a 5- to 7-membered heterocyclyl, a 3- to 6-membered heterocyclyl, a 4- to 6-membered heterocyclyl, a 5- or 6-membered heterocyclyl, a 4-membered heterocyclyl, a 5-membered heterocyclyl, a 6-membered heterocyclyl, or a 7-membered heterocyclyl. Heterocyclyl also includes ring systems in which the heterocyclyl groups described above are fused to one or more cycloalkyl groups, wherein the point of attachment is on the heterocyclyl ring, or the heterocyclyl groups described above are fused to one or more aryl or heteroaryl groups, wherein the point of attachment is on the heterocyclyl ring; and in such cases, the number of ring members continues to represent the number of ring members in the heterocyclyl ring system. The heterocyclyl group may further comprise a heterocyclyl ring as described above, wherein substituents on any non-adjacent carbon atoms or nitrogen atoms are linked to form a bridged ring, thereby together forming a polycyclic heteroalkane sharing two or more carbon atoms or nitrogen atoms. The heterocyclyl group may further comprise a heterocyclyl ring as described above, wherein substituents on the same carbon atom are linked to form a ring, thereby together forming a polycyclic heteroalkane sharing one carbon atom. Exemplary 3-membered heterocyclyl groups containing one heteroatom include, but are not limited to, aziridine, oxirane, and thiirane. Exemplary 4-membered heterocyclyl groups containing one heteroatom include, but are not limited to, azetidinyl, oxetane, and thiirane. Exemplary 5-membered heterocyclyl groups containing one heteroatom include, but are not limited to, tetrahydrofuranyl, dihydrofuranyl, tetrahydrothienyl, dihydrothienyl, pyrrolidinyl, dihydropyrrolyl, and pyrrolyl-2,5-dione. Exemplary 5-membered heterocyclic radicals containing two heteroatoms include, but are not limited to, pyrazolidinyl, dioxolanyl, oxasulfuranyl, disulfuranyl, and oxazolidin-2-one. Exemplary 5-membered heterocyclic radicals containing three heteroatoms include, but are not limited to, triazolinyl, oxadiazolinyl, and thiadiazolinyl. Exemplary 6-membered heterocyclic radicals containing one heteroatom include, but are not limited to, piperidinyl, tetrahydropyranyl, dihydropyridinyl, and tetrahydrothiopyranyl. Exemplary 6-membered heterocyclic radicals containing two heteroatoms include, but are not limited to, piperazinyl, morpholinyl, dithianyl, and dioxanyl.Exemplary 6-membered heterocyclyl groups containing three heteroatoms include but are not limited to triazinanyl. Exemplary 7-membered heterocyclyl groups containing one heteroatom include but are not limited to azepanyl, oxepane and thiepanyl. Exemplary 5-membered heterocyclyl groups fused to C6 aryl rings (also referred to herein as 5,6-bicyclic heterocyclyl) include but are not limited to indolinyl, isoindolinyl, dihydrobenzofuranyl, dihydrobenzothienyl, benzoxazolinonyl etc. Exemplary 6-membered heterocyclyl groups fused to C6 aryl rings (also referred to herein as 6,6-bicyclic heterocyclyl) include but are not limited to tetrahydroquinolinyl, tetrahydroisoquinolinyl etc. As long as valence allows, heterocyclyl further includes the heterocyclyl ring described above that shares one or two atoms with cycloalkyl, heterocyclyl, aryl or heteroaryl to form a bridged ring or spirocycle, wherein the shared atom can be a carbon atom or a nitrogen atom. Heterocyclyl further includes the heterocyclyl rings described above, which may be optionally substituted with one or more substituents, for example, 1 to 5 substituents, 1 to 3 substituents, or 1 substituent.

[0413] As used herein and unless otherwise indicated, "aryl" refers to a radical of a monocyclic or polycyclic 4n+2 aromatic ring system containing only carbon and hydrogen atoms. For example, "C 6-10 "Aryl" refers to a group of a monocyclic or polycyclic (e.g., bicyclic) 4n+2 aromatic ring system having 6-10 ring carbon atoms and zero heteroatoms (e.g., having 6 or 10 shared π electrons in the cyclic array). In some embodiments, an aryl group has six ring carbon atoms ("C6 aryl"; e.g., phenyl). In some embodiments, an aryl group has ten ring carbon atoms ("C 10 Aryl groups also include ring systems in which an aryl ring as described above is fused to one or more cycloalkyl or heterocyclyl groups, and the point of attachment is on the aryl ring, in which case the number of carbon atoms continues to refer to the number of carbon atoms in the aryl ring system. Aryl groups can be substituted or unsubstituted. Aryl groups can be substituted with one or more substituents, for example, with 1 to 5 substituents, 1 to 3 substituents, or 1 substituent.

[0414] As used herein and unless otherwise indicated, "heteroaryl" refers to a radical of a monocyclic or polycyclic 4n+2 aromatic ring system having at least one ring heteroatom. For example, a "5- to 14-membered heteroaryl" refers to a radical of a 5- to 14-membered monocyclic or bicyclic 4n+2 aromatic ring system having ring carbon atoms and ring heteroatoms (e.g., 1-4 ring heteroatoms) (e.g., having 6, 10, or 14 π electrons shared in a cyclic arrangement). In some embodiments, each heteroatom is independently selected from nitrogen, oxygen, and sulfur. In heteroaryl groups containing one or more nitrogen atoms, the point of attachment can be a carbon atom or a nitrogen atom as long as the valence permits. Heteroaryl bicyclic ring systems can include one or more heteroatoms in one or both rings. Heteroaryl also includes ring systems in which the heteroaryl ring described above is fused to one or more cycloalkyl or heterocyclyl groups, and the point of attachment is on the heteroaryl ring. In this case, the number of carbon atoms continues to represent the number of carbon atoms in the heteroaryl ring system. In some embodiments, the heteroaryl group is a 5- to 10-membered heteroaryl group, a 5- to 8-membered heteroaryl group, a 5- to 6-membered heteroaryl group, a 5-membered heteroaryl group, or a 6-membered heteroaryl group. Exemplary 5-membered heteroaryl groups containing one heteroatom include, but are not limited to, pyrrolyl, furanyl, and thienyl. Exemplary 5-membered heteroaryl groups containing two heteroatoms include, but are not limited to, imidazolyl, pyrazolyl, oxazolyl, isoxazolyl, thiazolyl, and isothiazolyl. Exemplary 5-membered heteroaryl groups containing three heteroatoms include, but are not limited to, triazolyl, oxadiazolyl (such as 1,2,4-oxadiazolyl), and thiadiazolyl. Exemplary 5-membered heteroaryl groups containing four heteroatoms include, but are not limited to, tetrazolyl. Exemplary 6-membered heteroaryl groups containing one heteroatom include, but are not limited to, pyridinyl or pyridonyl. Exemplary 6-membered heteroaryl groups containing two heteroatoms include, but are not limited to, pyridazinyl, pyrimidinyl, and pyrazinyl. Exemplary 6-membered heteroaryl groups containing three or four heteroatoms include, but are not limited to, triazinyl and tetrazinyl, respectively. Exemplary 7-membered heteroaryl groups containing one heteroatom include, but are not limited to, azacycloheptatrienyl, oxepantatrienyl, and thiepantatrienyl. Exemplary 5,6-bicyclic heteroaryl groups include, but are not limited to, indolyl, isoindolyl, indazolyl, benzotriazolyl, benzothiophenyl, isobenzothiophenyl, benzofuranyl, benzisofuranyl, benzimidazolyl, benzoxazolyl, benzisoxazolyl, benzoxadiazolyl, benzothiazolyl, benzisothiazolyl, benzthiadiazolyl, indolizinyl, and purinyl. Exemplary 6,6-bicyclic heteroaryl groups include, but are not limited to, naphthyridinyl, pteridinyl, quinolinyl, isoquinolinyl, cinnolinyl, quinoxalinyl, phthalazinyl, and quinazolinyl. Heteroaryl groups may be substituted or unsubstituted. The heteroaryl group may be substituted with one or more substituents, for example, with 1 to 5 substituents, 1 to 3 substituents, or 1 substituent.

[0415] "Hydroxyalkyl" refers to an alkyl group substituted with one or more hydroxy groups.

[0416] "Alkoxy" refers to the oxygen ether form of a straight or branched chain alkyl group, i.e., an -O-alkyl group. Similarly, "methoxy" refers to -O-CH3.

[0417] Divalent groups formed by removing another hydrogen from the groups defined above (such as alkyl, alkenyl, alkynyl, cycloalkyl, heterocyclyl, aryl and heteroaryl) are collectively referred to as "alkylene groups". Ring-forming groups such as cycloalkyl, heterocyclyl, aryl and heteroaryl are collectively referred to as "cyclyl groups".

[0418] As described herein and unless otherwise indicated, the term "optionally substituted" means unsubstituted or substituted. Substituents are independently selected, and substitution can be at any chemically acceptable position. As used herein, the term "substituted" means that a hydrogen atom is removed and replaced by a substituent. A single divalent substituent (e.g., oxo) can replace two hydrogen atoms. It should be understood that substitution at a given atom is limited by valence. Illustrative examples of substituents are substituents found in the exemplary compounds, groups, and embodiments disclosed herein, as well as halogen, deuterium, OH, CN, alkyl, haloalkyl, alkenyl, alkynyl, cycloalkyl, aryl, heterocyclyl, heteroaryl, heterocycloalkyl, cycloalkylalkyl, heterocyclylalkyl, or heteroarylalkyl. The substituent may optionally be further substituted with another substituent such as halogen, deuterium, OH, CN, alkyl, haloalkyl, alkoxy; cycloalkyloxy, aryloxy, heterocyclyloxy, heteroaryloxy, heterocycloalkyloxy, heterocyclylalkoxy, heteroaryloxy, oxo (═O), amino, alkylamino, cycloalkylamino, heteroarylamino, guanidino, amido, sulfonylamino, urea, nitrourea, oxime, hydroxyamino, alkoxyamino, hydrazine, azido, nitro, thio (—SH), sulfonyl, aminosulfonyl, acyl, formyl, carboxyl, ester, carbamate, amido, cyano or isocyanato.

[0419] All alkyl, alkenyl, alkynyl, cycloalkyl, heterocyclyl, aryl and heteroaryl groups, etc., as defined herein, are optionally substituted.

[0420] Exemplary substituents on carbon atoms include, but are not limited to, halogen, -CN, -NO2, N3, -SO2H, -SO3H, -OH, -OR aa 、-ON(R bb )2、-N(R bb )2、-N(R bb )3 + X - 、-N(OR cc )R bb 、-SH、-SR aa 、-SSR cc 、-C(=O)R aa、-CO2H、 -CHO、-C(OR cc )2、-CO2R aa 、-OC(=O)R aa 、-OCO2R aa 、-C(=O)N(R bb )2、-OC(=O)N(R bb )2、-NR bb C(=O)R a a 、-NR bb CO2R aa 、-NR bb C(=O)N(R bb )2、-C(=NR bb )R aa 、-C(=NR bb )OR aa 、-OC(=NR bb )R aa 、-OC(=NR bb )OR aa 、-C(=NR bb )N(R bb )2、-OC(=NR bb )N(R bb )2、-NR bb C(=NR bb )N(R bb )2、-C(=O)NR bb SO2R aa 、-NR bb SO2R aa 、-SO2N(R bb )2、-SO2R aa 、-SO2OR aa 、-OSO2R aa 、-S(=O)R aa 、-OS(=O)R aa 、-Si(R aa )3、-OSi(R aa )3、-C(=S)N(R bb )2、-C(=O)SR aa 、-C(=S)SR aa 、-SC(=S)SR aa 、-SC(=O)SR aa 、-OC(=O)SR aa 、-SC(=O)OR aa 、-SC(=O)R aa 、-P(=O)2R aa 、-OP(=O)2Raa 、-P(=O)(R aa )2、-OP(=O)(R aa )2、-OP(=O)(OR cc )2、-P(=O)2N(R bb )2、-OP(=O)2N(R bb )2、-P(=O)(NR bb )2、-OP(=O)(NR bb )2、-NR bb P(=O)(OR cc )2、-NR bb P(=O)(NR bb )2、-P(R cc )2、-P(R cc )3、-OP(R cc )2、-OP(R cc )3、-B(R aa )2、-B(OR cc )2, -BR aa (OR cc ), alkyl, haloalkyl, alkenyl, alkynyl, cycloalkyl, heterocyclyl, aryl and heteroaryl, wherein each of the alkyl, alkenyl, alkynyl, cycloalkyl, heterocyclyl, aryl and heteroaryl groups is independently replaced by 0, 1, 2, 3, 4 or 5 R dd or two geminal hydrogen atoms on the carbon atom are replaced by =O, =S, =NN(R bb )2, =NNR bb C(=O)R aa 、=NNR bb C(=O)OR aa 、=NNR bb S(=O)2R aa 、=NR bb NOR cc group replacement;

[0421] R aa Each of R is independently selected from alkyl, haloalkyl, alkenyl, alkynyl, cycloalkyl, heterocyclyl, aryl and heteroaryl, or R aa Two of the groups are combined to form a heterocyclyl or heteroaryl ring, wherein each of the alkyl, alkenyl, alkynyl, cycloalkyl, heterocyclyl, aryl, and heteroaryl groups is independently replaced by 0, 1, 2, 3, 4, or 5 R dd group substitution;

[0422] R bb Each of which is independently selected from hydrogen, -OH, -OR aa 、-N(R cc )2, -CN, -C(=O)Raa 、-C(=O)N(R cc )2, -CO2R aa 、-SO2R aa 、-C(=NR cc )OR aa 、-C(=NR cc )N(R cc )2、-SO2N(R cc )2, -SO2R cc 、-SO2OR cc 、-SOR aa 、-C(=S)N(R cc )2, -C(=O)SR cc 、-C(=S)SR cc 、-P(=O)2R aa 、-P(=O)(R aa )2、-P(=O)2N(R cc )2、-P(=O)(NR cc )2, alkyl, haloalkyl, alkenyl, alkynyl, cycloalkyl, heterocyclyl, aryl and heteroaryl, or two R bb The groups are combined to form a heterocyclyl or heteroaryl ring, wherein each of the alkyl, alkenyl, alkynyl, cycloalkyl, heterocyclyl, aryl, and heteroaryl groups is independently replaced by 0, 1, 2, 3, 4, or 5 R dd group substitution;

[0423] R cc Each of R is independently selected from hydrogen, alkyl, haloalkyl, alkenyl, alkynyl, cycloalkyl, heterocyclyl, aryl and heteroaryl, or two R cc The groups are combined to form a heterocyclyl or heteroaryl ring, wherein each of the alkyl, alkenyl, alkynyl, cycloalkyl, heterocyclyl, aryl, and heteroaryl groups is independently replaced by 0, 1, 2, 3, 4, or 5 R dd group substitution;

[0424] R dd Each of which is independently selected from halogen, -CN, -NO2, -N3, -SO2H, -SO3H, -OH, -OR ee 、-ON(R ff )2、-N(R ff )2、-N(R ff )3 + X - 、-N(OR ee )R ff 、-SH、-SR ee 、-SSR ee 、-C(=O)R ee 、-CO2H、-CO2Ree 、-OC(=O)R ee 、-OCO2R ee 、-C(=O)N(R ff )2、-OC(=O)N(R ff )2、-NR ff C(=O)R ee 、-NR ff CO2R ee 、-NR ff C(=O)N(R ff )2、-C(=NR ff )OR ee 、-OC(=NR ff )R ee 、-OC(=NR ff )OR ee 、-C(=NR ff )N(R ff )2、-OC(=NR ff )N(R ff )2、-NR ff C(=NR ff )N(R ff )2、-NR ff SO2R ee 、-SO2N(R ff )2, -SO2R ee 、-SO2OR ee 、-OSO2R ee 、-S(=O)R ee 、 -Si(R ee )3、-OSi(R ee )3、-C(=S)N(R ff )2, -C(=O)SR ee 、-C(=S)SR ee 、-SC(=S)SR ee 、-P(=O)2R ee 、-P(=O)(R ee )2、-OP(=O)(R ee )2、-OP(=O)(OR ee )2, alkyl, haloalkyl, alkenyl, alkynyl, cycloalkyl, heterocyclyl, aryl and heteroaryl, wherein each of alkyl, alkenyl, alkynyl, cycloalkyl, heterocyclyl, aryl and heteroaryl is independently replaced by 0, 1, 2, 3, 4 or 5 R gg Group substitution, or two geminal R dd Substituents may be combined to form =0 or =S;

[0425] R eeEach of R is independently selected from alkyl, haloalkyl, alkenyl, alkynyl, cycloalkyl, aryl, heterocyclyl and heteroaryl, wherein each of alkyl, alkenyl, alkynyl, cycloalkyl, heterocyclyl, aryl and heteroaryl is independently replaced by 0, 1, 2, 3, 4 or 5 R gg group substitution;

[0426] R ff Each of R is independently selected from hydrogen, alkyl, haloalkyl, alkenyl, alkynyl, cycloalkyl, heterocyclyl, aryl and heteroaryl, or two R ff The groups are combined to form a heterocyclyl or heteroaryl ring, wherein each of the alkyl, alkenyl, alkynyl, cycloalkyl, heterocyclyl, aryl, and heteroaryl groups is independently replaced by 0, 1, 2, 3, 4, or 5 R gg group substitution;

[0427] R gg Each of which is independently selected from halogen, -CN, -NO2, N3, -SO2H, -SO3H, -OH, -OC 1-6 Alkyl, -ON(C 1-6 Alkyl)2, -N(C 1-6 Alkyl)2, -N(C 1-6 Alkyl)3 + X - 、-NH(C 1-6 Alkyl)2 + X - 、-NH2(C 1-6 alkyl) + X - 、-NH3 + X - 、-N(OC 1-6 Alkyl)(C 1-6 Alkyl), -N(OH)(C 1-6 Alkyl), -NH(OH), -SH, -SC 1-6 Alkyl, -SS(C 1-6 alkyl), -C(=O)(C 1-6 alkyl), -CO2H, -CO2(C 1-6 alkyl), -OC(=O)(C 1-6 Alkyl), -OCO2(C 1-6 alkyl), -C(=O)NH2, -C(=O)N(C 1-6 alkyl)2, -OC(=O)NH(C 1-6 alkyl), -NHC(=O)(C 1-6 Alkyl), -N(C 1-6 alkyl)C(=O)(C 1-6 Alkyl), -NHCO2(C 1-6alkyl), -NHC(=O)N(C 1-6 Alkyl)2, -NHC(=O)NH(C 1-6 alkyl), -NHC(=O)NH2, -C(=NH)O(C 1-6 alkyl), -OC(=NH)(C 1-6 alkyl), -OC(=NH)OC 1-6 Alkyl, -C(=NH)N(C 1-6 Alkyl)2, -C(=NH)NH(C 1-6 alkyl), -C(=NH)NH2, -OC(=NH)N(C 1-6 Alkyl)2, -OC(NH)NH(C 1-6 alkyl), -OC(NH)NH2, -NHC(NH)N(C 1-6 Alkyl)2, -NHC(=NH)NH2, -NHSO2(C 1-6 Alkyl), -SO2N(C 1-6 Alkyl)2, -SO2NH(C 1-6 alkyl), -SO2NH2, -SO2C 1-6 Alkyl, -SO2OC 1-6 Alkyl, -OSO2C 1-6 Alkyl, -SOC 1-6 Alkyl, -Si(C 1-6 alkyl)3, -OSi(C 1-6 alkyl)3, -C(=S)N(C 1-6 alkyl)2、C(=S)NH(C 1-6 alkyl), C(=S)NH2, -C(=O)S(C 1-6 alkyl), -C(=S)SC 1-6 Alkyl, -SC(=S)SC 1-6 Alkyl, -P(=O)2(C 1-6 alkyl), -P(=O)(C 1-6 alkyl)2, -OP(=O)(C 1-6 alkyl)2, -OP(=O)(OC 1-6 Alkyl)2, C 1-6 Alkyl, C 1-6 Halogenated alkyl, C2-C6 alkenyl, C2-C6 alkynyl, C3-C7 cycloalkyl, C6-C 10 Aryl, C3-C7 heterocyclic, C5-C 10 heteroaryl; or two geminal R gg Substituents may combine to form =O or =S; where X - It is a counter ion.

[0428] Exemplary substituents on nitrogen atoms include, but are not limited to, hydrogen, -OH, -OR aa 、-N(R cc )2, -CN, -C(=O)R aa 、-C(=O)N(R cc )2, -CO2R aa 、-SO2R aa 、-C(=NR bb )R aa 、-C(=NR cc )OR aa 、-C(=NR cc )N(R cc )2、-SO2N(R cc )2, -SO2R cc 、-SO2OR cc 、-SOR aa 、-C(=S)N(R cc )2, -C(=O)SR cc 、-C(=S)SR cc 、-P(=O)2R aa 、-P(=O)(R aa )2、-P(=O)2N(R cc )2、-P(=O)(NR cc )2, alkyl, haloalkyl, alkenyl, alkynyl, cycloalkyl, heterocyclyl, aryl and heteroaryl, or two R cc The groups combine to form a heterocyclyl or heteroaryl ring, wherein each of the alkyl, alkenyl, alkynyl, cycloalkyl, heterocyclyl, aryl, and heteroaryl groups is independently replaced by 0, 1, 2, 3, 4, or 5 R dd group substituted, and wherein R aa 、R bb 、R cc and R dd As described herein.

[0429] "Nucleic acid" refers to single-stranded or double-stranded deoxyribonucleic acid (DNA) or ribonucleic acid (RNA) molecules and hybrid molecules thereof. Examples of nucleic acid molecules include, but are not limited to, messenger RNA (mRNA), microRNA (miRNA), small interfering RNA (siRNA), self-amplifying RNA (saRNA), and antisense oligonucleotides (ASOs). The nucleic acid can be further chemically modified, and the chemical modifier is selected from one or a combination of the following substances: pseudouridine, N1-methyl-pseudouridine, 5-methoxyuridine, and 5-methylcytosine. The mRNA molecule contains a protein coding region and may further contain an expression regulatory sequence. Typical expression regulatory sequences include, but are not limited to, a 5' cap, a 5' untranslated region (5'UTR), a 3' untranslated region (3'UTR), a polyadenylic acid sequence (PolyA), and a miRNA binding site.

[0430] As used herein, the term "pKa" refers to the negative logarithm (p) of the acid dissociation constant (Ka) of an acid, and is equal to the pH value when there are equal concentrations of acid and its conjugate base form in a solution. As used herein, the term "pKa" can be measured using water or dimethyl sulfoxide as a solvent. Observations previously reported as pKa using water as a solvent can be used as pKa as used herein. In some embodiments, pKa can be determined experimentally, such as titration experiments using hydrochloric acid or sodium hydroxide. In some embodiments, pKa is determined by 2-(p-toluidinyl)naphthalene-6-sulfonic acid (TNS) fluorescence method.

[0431] As used herein, the term "ionizable" or "ionizable group" refers to a chemical group that is ionized or capable of ionization. An ionizable group can exist as a neutral group, a positively charged group (cationic group), or a negatively charged group (anionic group). As used herein, a "charged moiety" is a chemical moiety that carries an electric charge, for example, monovalent (+1, or -1), divalent (+2, or -2), trivalent (+3, or -3), etc.

[0432] The term "treat" as used herein relates to reversing, alleviating, or inhibiting the progression of, or preventing, the disorder or condition to which the term applies, or reversing, alleviating, or inhibiting the progression of, or preventing one or more symptoms of, such a disorder or condition. The noun "treat" as used herein relates to the action of treating (verb), and treating (verb) is as just defined.

[0433] As used herein, the term "pharmaceutically acceptable salt" refers to carboxylate and amino acid addition salts of the compounds of the present disclosure that are suitable for contact with the tissues of patients within the scope of sound medical judgment and do not produce undue toxicity, irritation, allergic response, or the like. They are commensurate with a reasonable benefit / risk ratio and are effective for their intended use. Where possible, the term includes zwitterionic forms of the compounds of the present disclosure.

[0434] Pharmaceutically acceptable base addition salts are formed from metals or amines, such as alkali metal and alkaline earth metal hydroxides or organic amines. Examples of metals used as cations include sodium, potassium, magnesium, calcium, and the like. Examples of suitable amines are N,N'-dibenzylethylenediamine, chloroprocaine, choline, diethanolamine, ethylenediamine, N-methylglucamine, and procaine.

[0435] Base addition salts of acidic compounds can be prepared by contacting the free acid form with a sufficient amount of the desired base in a conventional manner to form a salt. The free acid can be regenerated by contacting the salt form with an acid in a conventional manner and then isolating the free acid. The free acid form differs from its salt form in physical properties, such as solubility in polar solvents. However, for the purposes of this disclosure, the salt is still equivalent to its free acid.

[0436] Salt can be prepared by inorganic acid, and the inorganic acid comprises sulfate, pyrosulfate, bisulfate, sulfite, bisulfite, nitrate, phosphate, monohydrogen phosphate, dihydrogen phosphate, metaphosphate, pyrophosphate, chloride, bromide and iodide.The example of inorganic acid comprises hydrochloric acid, nitric acid, sulfuric acid, hydrobromic acid, hydroiodic acid, phosphoric acid etc.Representative salt comprises hydrobromide, hydrochloride, sulfate, bisulfate, nitrate, acetate, oxalate, valerate, oleate, palmitate, stearate, laurate, borate, benzoate, lactate, phosphate, toluenesulfonate, citrate, maleate, fumarate, succinate, tartrate, naphthalene dicarboxylate, methylsulfonate, glucoheptanoate, lactobionate, lauryl sulfonate, isethionate etc. Salts can also be prepared from organic acids including aliphatic monocarboxylic acids and dicarboxylic acids, phenyl-substituted alkanoic acids, hydroxyalkanoic acids, alkanedioic acids, aromatic acids, aliphatic and aromatic sulfonic acids, and the like. Representative salts include acetate, propionate, octanoate, isobutyrate, oxalate, malonate, succinate, suberate, sebacate, fumarate, maleate, mandelate, benzoate, chlorobenzoate, methylbenzoate, dinitrobenzoate, naphthoate, benzenesulfonate, toluenesulfonate, phenylacetate, citrate, lactate, maleate, tartrate, methanesulfonate, and the like. Pharmaceutically acceptable salts can include cations based on alkali metals and alkaline earth metals (e.g., sodium, lithium, potassium, calcium, magnesium, and the like), as well as non-toxic ammonium, quaternary ammonium, and amine cations, including but not limited to ammonium, tetramethylammonium, tetraethylammonium, methylamine, dimethylamine, trimethylamine, triethylamine, ethylamine, and the like. Also included are salts of amino acids such as arginine salts, gluconates, galacturonates, and the like (see, for example, Berge SM et al., "Pharmaceutical Salts," J. Pharm. Sci., 1977; 66: 1-19 for reference).

[0437] "Subjects" contemplated for administration include, but are not limited to, humans (e.g., males or females of any age group, e.g., pediatric subjects (e.g., infants, children, adolescents) or adult subjects (e.g., young, middle-aged, or elderly) and / or non-human animals, such as mammals, e.g., primates (e.g., cynomolgus monkeys, rhesus monkeys), cows, pigs, horses, sheep, goats, rodents, cats, and / or dogs. In some embodiments, the subject is a human. In some embodiments, the subject is a non-human animal. The terms "human," "patient," and "subject" are used interchangeably herein.

[0438] "Disease," "disorder," and "condition" are used interchangeably herein.

[0439] Unless otherwise indicated, the term "treat," as used herein, includes an effect on a subject having a particular disease, disorder, or condition that reduces the severity of the disease, disorder, or condition, or delays or slows the progression of the disease, disorder, or condition ("therapeutic treatment"). The term also includes an effect produced before a subject begins to suffer from a particular disease, disorder, or condition ("prophylactic treatment").

[0440] Typically, the "effective amount" of a pharmaceutical composition refers to an amount sufficient to elicit a target biological response. As will be appreciated by those skilled in the art, the effective amount of a pharmaceutical composition of the present disclosure may vary depending on factors such as the desired biological endpoint, the pharmacokinetics of the pharmaceutical composition, the disease being treated, the mode of administration, and the age, health status, and symptoms of the subject. An effective amount includes a therapeutically effective amount and a prophylactic effective amount. Unless otherwise indicated, a "therapeutically effective amount" of a pharmaceutical composition as used herein is an amount sufficient to provide a therapeutic benefit in the process of treating a disease, disorder, or condition, or to delay or minimize one or more symptoms associated with the disease, disorder, or condition. A therapeutically effective amount of a pharmaceutical composition refers to an amount of a therapeutic agent that provides a therapeutic benefit in the treatment of a disease, disorder, or condition when used alone or in combination with other therapies. The term "therapeutically effective amount" may include an amount that improves overall treatment, reduces or avoids the symptoms or causes of a disease or condition, or enhances the therapeutic effect of other therapeutic agents.

[0441] Unless otherwise indicated, the term "steroid compound" or "cationic steroid compound" as used herein refers to any steroid compound comprising at least one cationic portion. As used herein, the term "steroid" is a class of compounds having a tetracyclic 17-carbon ring structure, which may further comprise one or more substitutions, including but not limited to alkyl groups, alkoxy groups, hydroxyl groups, oxo groups, acyl groups, or double bonds between two or more carbon atoms. As used herein, the term "steroid" also includes steroid esters and sterols. As used herein, the term "steroid" also includes compounds belonging to or related to the following illustrative compound families: corticosteroids, mineralocorticoids, and sex steroids (e.g., androgens or estrogens or anti-androgens and anti-estrogens molecules). Non-limiting examples of steroids include cholesterol, cholestane, campesterol, sitosterol, sitostanol, β-sitosterol, stigmasterol, stigmasterol, rapeseed sterol, avenasterol, ergosterol, ursolic acid, tocopherol, photosterol, or derivatives thereof. In one embodiment, the cationic steroid compound is an ionizable steroid compound or a zwitterionic steroid compound that becomes positively charged at a certain pH. In one embodiment, the cationic steroid compound is a permanent cationic steroid compound. In one embodiment, the cationic steroid compound is a compound described in Section 0.

[0442] Unless otherwise indicated, the term "biodegradable" as used herein refers to a substance that is degraded under physiological conditions. In some embodiments, a biodegradable substance is a substance that is decomposed by an organelle. In some embodiments of any aspect, a biodegradable substance is a substance that is decomposed by a chemical process. As used herein, the term "biodegradable group" refers to a group comprising one or more bonds that can undergo a bond cleavage reaction in a biological environment, such as in an organism, organ, tissue, cell, or organelle. For example, a biodegradable group can be metabolized (for example, by hydrolysis) by the body of a mammal (such as a human). Some groups containing biodegradable bonds include, for example, but are not limited to, esters, dithiols, and oximes. Non-limiting examples of biodegradable groups are -OC(O)-, -C(O)O-, -SC(O)-, -C(O)S-, -OC(S)-, -C(S)O-, -SS-, -C(R cc )=N-、-N=C(R cc )-、-C(R cc )=NO-、-ON=C(R cc )-、-C(O)(NR cc )-、-N(R cc )C(O)-、-C(S)(NR cc )-、-N(R cc )C(O)-、-N(R cc )C(O)N(R cc )-、-OC(O)O-、-OSi(R cc )2O-、-C(O)(CR cc R cc )C(O)O- or -OC(O)(CR cc R cc )C(O)-. The term "biodegradable lipid" refers to a lipid having one or more biodegradable groups in the middle or distal end of the lipid portion (e.g., hydrophobic chain) of the lipid. Without being bound by theory, incorporating biodegradable groups into lipids may result in faster metabolism and removal of the lipid from the body after payload delivery.

[0443] Unless otherwise indicated, the term "structured lipid" refers to a neutral sterol, and also refers to a lipid containing a neutral sterol portion. Without being bound by theory, incorporating structured lipids into lipid nanoparticles can help alleviate the aggregation of other lipids in the particle. Structured lipids can be selected from a group including but not limited to cholesterol, fecosterol, sitosterol, ergosterol, campesterol, stigmasterol, rapeseed sterol, tomatine, tomatine, ursolic acid, α-tocopherol, hopanes, plant sterols, or a mixture thereof. In some embodiments, the structured lipid is a sterol. As defined herein, "sterol" is a subgroup of the steroids consisting of steroid alcohols. In certain embodiments, the structured lipid is cholesterol. In certain embodiments, the structured lipid is an analog of cholesterol. In certain embodiments, the structured lipid is α-tocopherol. Unless otherwise indicated, "structured lipid" as used herein is not a "steroid compound" or "cationic steroid compound" as used herein. In one embodiment, a "structured lipid" as used herein does not comprise a cationic moiety or an ionizable moiety. In one embodiment, a "structured lipid" as used herein does not comprise a quaternary ammonium group or a tertiary amine group.

[0444] Unless otherwise indicated, the term "polymer-conjugated lipid" as used herein refers to a polymer (e.g., polyethylene glycol (PEG), polypropylene glycol, polyvinyl pyrrolidone, or poly(N-(2-hydroxypropyl)methacrylamide)) conjugated to a lipid. For example, PEG can be conjugated to myristoyl diglyceride to generate DMG-PEG-2000. Alternatively, PEG can be conjugated to DSPE to generate DSPE-PEG-2000. In one embodiment, the polymer-conjugated lipid is a PEGylated lipid. PEGylated lipids can incorporate various functionalized PEG end groups, including amine, carboxylic acid, azide, aldehyde, thiol, and hydroxyl moieties. Without being bound by theory, polymer-conjugated lipids improve circulation time, drug stability, suitability for different routes of administration, and facilitate targeted drug delivery.

[0445] 5.2 Lipid Nanoparticles

[0446] In one aspect, a lipid nanoparticle (LNP) is provided herein. In one embodiment, the LNP comprises a steroid compound (steroid compound, chapter 0). In one embodiment, the LNP comprises an ionizable lipid (chapter 0). In one embodiment, the LNP comprises a steroid compound (chapter 0) and an ionizable lipid (chapter 0). In one embodiment, the LNP further comprises a phospholipid lipid (chapter 0). In one embodiment, the LNP does not comprise a phospholipid lipid. In one embodiment, the LNP further comprises a structural lipid (chapter 0). In one embodiment, the LNP further comprises a polymer-conjugated lipid (chapter 0). In one embodiment, the LNP further comprises a payload (chapter 0).

[0447] In one embodiment, the lipid nanoparticles described herein are not liposomes. In one embodiment, the lipid nanoparticles described herein are substantially free of an aqueous lumen, or have an lumen that is less than 10% of the total volume of the nanoparticle. In one embodiment, the lipid nanoparticles described herein comprise a unilamellar lipid membrane. In one embodiment, the lipid nanoparticles described herein comprise smaller particles within the interior of the lipid nanoparticle, wherein the smaller particles are associated with a payload described herein.

[0448] In one embodiment, the present invention provides a lipid nanoparticle for delivering or expressing a payload in a subject's spleen. In one embodiment, the amount of the payload delivered or expressed in the subject's spleen is higher than in the subject's liver. In one embodiment, the payload is delivered by systemic administration.

[0449] In one embodiment, the lipid nanoparticles contain a steroid compound and an ionizable lipid.

[0450] In one embodiment, the molar percentage of the steroid compound to the total lipids is 3-15 mol%, preferably 5-10 mol%, for example 3 mol%, 4 mol%, 5 mol%, 6 mol%, 7 mol%, 8 mol%, 9 mol%, 10 mol%, 11 mol%, 12 mol%, 13 mol%, 14 mol% or 15 mol%.

[0451] In one embodiment, the molar percentage of the ionizable lipid is 35-70 mol%, more preferably 40-60 mol%, such as 35 mol%, 40 mol%, 45 mol%, 50 mol%, 55 mol% or 60 mol%.

[0452] In one embodiment, the spleen targeting lipid nanoparticles further comprise a phospholipid.

[0453] In a preferred embodiment, the molar percentage of the phospholipids is 5-30 mol%, preferably 5-20 mol%, more preferably 5-15 mol%, for example 5 mol%, 6 mol%, 7 mol%, 8 mol%, 9 mol%, 10 mol%, 11 mol%, 12 mol%, 13 mol%, 14 mol%, 15 mol%, 16 mol%, 17 mol%, 18 mol%, 19 mol% or 20 mol%.

[0454] In one embodiment, the spleen-targeted lipid nanoparticles further comprise structural lipids, such as cholesterols.

[0455] In a preferred embodiment, the molar percentage of the structured lipid is 10-80 mol%, more preferably 15-70 mol%, more preferably 30-50 mol%, more preferably 30-40 mol%, for example 30 mol%, 35 mol%, 40 mol%, 45 mol%, 50 mol%, 55 mol% or 60 mol%.

[0456] In one embodiment, the spleen-targeted lipid nanoparticles further comprise a polymer-conjugated lipid, such as a pegylated lipid.

[0457] In a preferred embodiment, the molar percentage of the polymer-conjugated lipid is 0.25-10 mol%, more preferably 0.5-5 mol%, more preferably 0.5-3 mol%, for example 0.5 mol%, 0.6 mol%, 0.7 mol%, 0.8 mol%, 0.9 mol%, 1.0 mol%, 1.1 mol%, 1.2 mol%, 1.3 mol%, 1.4 mol%, 1.5 mol%, 1.6 mol%, 1.7 mol%, 1.8 mol%, 1.9mol%, 2.0mol%, 2.1mol%, 2.2mol%, 2.3mol%, 2.4mol%, 2.5mol%, 2.6mol%, 2.7mol%, 2.8mol%, 2.9mol%, 3.0 mol%, 3.1 mol%, 3.2 mol%, 3.3 mol%, 3.4 mol%, 3.5 mol%, 3.6 mol%, 3.7 mol%, 3.8 mol%, 3.9 mol%, 4.0 mol%.

[0458] In one embodiment, the average particle size of the spleen-targeted lipid nanoparticles is 50-800 nm, more preferably 50-400 nm, more preferably 80-200 nm, for example 80, 90, 100, 110, 120, 130, 140, 150, 160, 170, 180, 190 or 200 nm.

[0459] In one embodiment, the lipid nanoparticles contain: (1) a steroid compound; (2) an ionizable lipid; (3) a phospholipid compound; (4) a structural lipid; and (5) a polymer-conjugated lipid. In a more preferred embodiment, the lipid nanoparticles further contain a payload.

[0460] In a preferred embodiment, the spleen-targeted lipid nanoparticles include the following components in molar percentages:

[0461] 6) steroid compound 3-15 mol%;

[0462] 7) ionizable lipids 35-70 mol%;

[0463] 8) 5-30 mol% phospholipids;

[0464] 9) structural lipids 10-80 mol%; and

[0465] 10) Polymer-conjugated lipid 0.25-10 mol%.

[0466] In a preferred embodiment, the spleen-targeted lipid nanoparticles include the following components in molar percentages:

[0467] 6) steroid compound 5-10 mol%;

[0468] 7) ionizable lipids 40-60 mol%;

[0469] 8) 5-20 mol% phospholipids;

[0470] 9) structural lipids 15-70 mol%; and

[0471] 10) Polymer-conjugated lipid 0.5-5 mol%.

[0472] In a preferred embodiment, the spleen-targeted lipid nanoparticles include the following components in molar percentages:

[0473] 6) steroid compound 5-10 mol%;

[0474] 7) ionizable lipids 40-60 mol%;

[0475] 8) 5-15 mol% phospholipids;

[0476] 9) structural lipids 30-50 mol%; and

[0477] 10) Polymer conjugated lipid 0.5-3 mol%.

[0478] In a particularly preferred embodiment, the spleen-targeted lipid nanoparticles include the following components in molar percentages:

[0479] 6) steroid compound 5 mol%;

[0480] 7) ionizable lipids 50 mol%;

[0481] 8) phospholipids 10 mol%;

[0482] 9) structural lipids 33.5 mol%; and

[0483] 10) Polymer-conjugated lipid 1.5 mol%.

[0484] In one embodiment, the particle size distribution (PDI) of the lipid nanoparticles is no greater than 0.2, preferably no greater than 0.15.

[0485] In one embodiment, the ratio of the amount of the lipid nanoparticles distributed to the spleen to that distributed to the liver after administration is greater than 1, more preferably greater than 2, more preferably greater than 3, more preferably greater than 4, more preferably greater than 5, more preferably greater than 10, more preferably greater than 20, more preferably greater than 30, more preferably greater than 40, more preferably greater than 50, and more preferably greater than 100.

[0486] In one embodiment, the ratio of the lipid nanoparticles distributed to the spleen and the lungs after administration is greater than 1, more preferably greater than 2, more preferably greater than 3, more preferably greater than 4, more preferably greater than 5, more preferably greater than 10, more preferably greater than 20, more preferably greater than 30, more preferably greater than 40, more preferably greater than 50, and more preferably greater than 100.

[0487] 5.2.1. Steroid compounds

[0488] In one embodiment, the lipid nanoparticles comprise a steroid compound comprising one or more cationic groups.

[0489] In one embodiment, the cationic group of the steroid compound is a tertiary amine group, a quaternary ammonium group, an ammonium group (NH3 + ), a primary amine group (NH2), an imidazole group, a pyridine group, a pyrrole group, a pyrrolidine group, a pyrazole group, an isothiazole group, an isoxazole group, an oxadiazine group, an oxazoline group, a dithiourea group, an isotriazole group, a tetrazole group, a pentazole group, an indole group, a dihydroindole group, an isoxazole group, a pyrimidine group, a pyrazine group, a quinazoline group, a piperazine group, a piperidine group, a morpholine group, a pyran group, a quinoxaline group, a quinoline group, a quinolinone group, an isoquinoline group, an amidine group, a guanidine group or a urea group.

[0490] In one embodiment, the cationic group of the steroid compound is a quaternary ammonium group. In one embodiment, the cationic group is a tertiary amine group. In one embodiment, the steroid compound comprises a quaternary ammonium group and a β-sitosterol moiety. In one embodiment, the cationic steroid compound comprises a tertiary amine group and a β-sitosterol moiety. In one embodiment, the cationic group is an amidine group, a guanidine group, or a urea group.

[0491] In one embodiment, the steroid compound is a permanently cationic steroid compound (i.e., positively charged regardless of pH). In one embodiment, the steroid compound is an ionizable cationic steroid compound (e.g., positively charged at a certain pH). In one embodiment, the steroid compound is a zwitterionic steroid compound.

[0492] In one embodiment, the steroid compound comprises a Z1 moiety comprising three fused cyclohexyl rings and one fused cyclopentyl ring, wherein one or more CC bonds in the rings are optionally replaced by a C=C double bond. In one embodiment, Z1 comprises an optionally substituted cyclopentanepolyhydrophenanthrene moiety.

[0493] In one embodiment, the steroid compound comprises a Z1 moiety, and wherein said Z1 is or a stereoisomer or mixture of stereoisomers thereof, wherein one or more ring C—C bonds in Z1 are independently optionally replaced by C═C bonds where valence permits, wherein Z1 is optionally replaced by one or more rings selected from hydroxy, halogen and C 1-14 Alkyl or C 1-14 wherein the alkyl group is optionally substituted with one or more hydroxyl, nitro, cyano, halogen, C 1-6 Alkoxy, C2-C6 alkenyl, C2-C6 alkynyl, C3-C8 cycloalkyl, C6-C 10 Z1 is substituted with aryl, 5- to 10-membered heteroaryl, or 3- to 8-membered heterocyclyl, and wherein the linkage in Z1 is toward the remainder of the steroid compound.

[0494] In one embodiment, the steroid compound comprises a Z1 moiety, wherein the Z1 moiety is a monovalent free radical of cholesterol, cholestane, campesterol, sitosterol, sitostanol, β-sitosterol, stigmasterol, stigmasterol, brassicasterol, avenasterol, ergosterol, ursolic acid, tocopherol, or a stereoisomer, a mixture of stereoisomers, or a pharmaceutically acceptable salt thereof.

[0495] In one embodiment, the pKa of the steroid compound is between 5 and 8. In one embodiment, the pKa of the steroid compound is between 5.5 and 6.5. In one embodiment, the pKa of the steroid compound is greater than 8. In one embodiment, the pKa of the steroid compound is greater than 10. In one embodiment, the pKa of the steroid compound is greater than 10.

[0496] In one embodiment, the steroid compound is a compound of formula (C): M - V1-[-(R 20 ) a1 -Y1] n1 -Z1 (C)

[0497] or a stereoisomer, a mixture of stereoisomers, or a pharmaceutically acceptable salt thereof, wherein:

[0498] Z1 is an optionally substituted steroid;

[0499] Each instance of Y1 is independently absent or -C(O)O-, -O-, -NH-, -SC(O)O-, -OC(O)NR 2a -、-NR 2a C(O)NR 2a -、-OC(O)S-、-OC(O)O-、-NR 2a C(O)O-, -OC(O)-, -SC(O)-, -C(O)S-, -NR 2a -、-C(O)NR 2a -、-NR 2a C(O)-、-NR 2a C(O)S-、-SC(O)NR 2a -, -C(O)-, -OC(S)-, -C(S)O-, -OC(S)NR 2a -、-NR 2a C(S)O-, -SS-, or -S(O) 0-2 -, where the leftward connection is in the direction of V1 and the rightward connection is in the direction of Z1;

[0500] R 2a Each instance of is independently H, C 1-10 Alkyl, C 3-14 cycloalkyl or 3 to 14 membered heterocyclyl, and wherein the alkyl, cycloalkyl and heterocyclyl are independently optionally replaced by C 1-10 Alkyl, L 2b -OR 2b , Lb-SR 2b 、L 2b -NR 2b R'2b or L 2b N + R 2b R' 2b R” 2b replace;

[0501] R 20 Each example of is independently -(CR 201 R 202 )-, where R 201 and R 202 Each example is independently H, OH, alkyl, alkoxy, -L 2b -NR 21 R 22 or -L 2b -N + R 21 R 22 R 23 ;

[0502] a1 and n1 are each independently 0, 1, 2, 3, 4, 5, 6, 7, 8, 9 or 10;

[0503] V1 is NR 21 R 22 、N + R 21 R 22 R 23 , or an optionally substituted nitrogen-containing heterocyclic group selected from the group consisting of imidazole, pyridine, pyrrole, pyrrolidine, pyrazole, isothiazole, isoxazole, oxadiazine, oxazoline, dithioureaazole, isotriazole, tetrazole, pentazole, indole, indoline, isoxazole, pyrimidine, pyrazine, quinazoline, piperazine, piperidine, morpholine, pyran, quinoxaline, quinoline, quinolinone and isoquinoline, or a nitrogen-containing non-heterocyclic group selected from amidine, guanidine or urea;

[0504] M is absent or is an anion selected from the group consisting of chloride, bromide, iodide, sulfate, nitrate, perchlorate, formate, phosphate, pyrophosphate, citrate, sulfonate, methanesulfonate, trifluoromethanesulfonate, and trifluoroacetate;

[0505] R 21 、R 22 、R 23 Each instance of is independently H, C 1-8 Alkyl, -L 2b -OR 2b 、-L 2b -SR 2b or-L2c-NR 2b R' 2b R” 2b ;

[0506] L2b Does not exist or C 1-10 an alkylene group; and

[0507] R 2b , R' 2b and R” 2b Each of is independently H or C 1-10 alkyl.

[0508] In one embodiment, a1 is 3, 4, 5, 6, 7, 8, 9 or 10.

[0509] In one embodiment, n1 is 0, 1, 2, 3, 4, 5, 6, 7, 8, 9 or 10.

[0510] In one embodiment, the steroid compound is a compound of formula (C1): M - V1-(R 20 ) 2-8 -Y1-Z1 (C1)

[0511] In one embodiment, the steroid compound is a compound of formula (C2): M - V1-(R 20 ) 2-6 -Y1-(R 20 ) 2-6 -Y1-Z1 (C2)

[0512] In one embodiment, R20 is -(CR 201 R 202 )-, where R 201 and R 202 Each example is independently H, OH, alkyl, alkoxy, -L 2b -NR 21 R 22 or -L 2b -N + R 21 R 22 R 23 In one embodiment, R20 is -(CR 201 R 202 )-, where R 201 and R 202 Each instance of is independently H, C 1-6 Alkyl, -NR 21 R 22 or -N + R 21 R 22 R 23 ;

[0513] In one embodiment, the steroid compound is a compound of formula (I):

[0514] or a stereoisomer, a mixture of stereoisomers, or a pharmaceutically acceptable salt thereof, wherein:

[0515] Is a single bond or a double bond;

[0516] Y is -C(O)O-, -O-, -NH-, -SC(O)O-, -OC(O)NR 2a -、-NR 2a C(O)NR 2a -、-OC(O)S-、-OC(O)O-、-NR 2a C(O)O-, -OC(O)-, -SC(O)-, -C(O)S-, -NR 2a -、-C(O)NR 2a -、-NR 2a C(O)-、-NR 2a C(O)S-、-SC(O)NR 2a -, -C(O)-, -OC(S)-, -C(S)O-, -OC(S)NR 2a -、-NR 2a C(S)O- or -S(O) 0-2 -, and wherein the leftward connection is in the direction of V1;

[0517] Y1 is absent or selected from -C(O)O-, -O-, -NH-, -SC(O)O-, -OC(O)NR 2a -、-NR 2a C(O)NR 2a -、-OC(O)S-、-OC(O)O-、-NR 2a C(O)O-, -OC(O)-, -SC(O)-, -C(O)S-, -NR 2a -、-C(O)NR 2a -、-NR 2a C(O)-、-NR 2a C(O)S-、-SC(O)NR 2a -, -C(O)-, -OC(S)-, -C(S)O-, -OC(S)NR 2a -、-NR 2a C(S)O- or -S(O) 0-2 -, and wherein the leftward connection is in the direction of V1;

[0518] R 2a Each instance of is independently H, C 1-10Alkyl, C 3-14 cycloalkyl or 3 to 14 membered heterocyclyl, and wherein the alkyl, cycloalkyl and heterocyclyl are independently optionally replaced by C 1-10 Alkyl, L 2b -OR 2b , Lb-SR 2b 、L 2b -NR 2b R' 2b 、L 2b N + R 2b R' 2b R” 2b 、L 2b -OC(O)R 2b or L 2b -C(O)OR 2b replace;

[0519] R 201 、R 202 、R 203 、R 204 Each example is independently H, OH, alkyl, alkoxy, -L 2b -NR 21 R 22 、-L 2b -N + R 21 R 22 R 23 、L 2b -OC(O)R 21 or L 2b -C(O)OR 21 ;

[0520] m is an integer from 1 to 12;

[0521] m1 is an integer from 0 to 12;

[0522] V1 is NR 21 R 22 、N + R 21 R 22 R 23 , imidazole, pyridine, pyrrole, pyrrolidine, pyrazole, isothiazole, isoxazole, oxadiazine, oxazoline, dithioureaazole, isotriazole, tetrazole, pentazole, indole, indoline, isoxazole, pyrimidine, pyrazine, quinazoline, piperazine, piperidine, morpholine, pyran, quinoxaline, quinoline, quinolinone, isoquinoline, amidine, guanidine or urea;

[0523] M is absent or is one or more anions independently selected from chloride, bromide, iodide, sulfate, nitrate, perchlorate, formate, citrate, sulfonate, methanesulfonate, trifluoromethanesulfonate, trifluoroacetate, pyrosulfate, bisulfate, sulfite, bisulfite, phosphate, pyrophosphate, monohydrogen phosphate, dihydrogen phosphate, acetate, propionate, decanoate, octanoate, acrylate, isobutyrate, hexanoate, heptanoate, oxalate, malonate, succinate, fumarate, maleate, benzoate, phthalate, phenylacetate, lactate, glycolate, tartrate, mandelate, methanesulfonate, gluconate, aspartate, or glutamate;

[0524] R 21 、R 22 、R 23 Each instance of is independently H, optionally substituted C 1-8 Alkyl, -L 2b -OR 2b 、-L 2b -SR 2b 、-L 2b -NR 2b R' 2b R” 2b 、L 2b -OC(O)R 2b or L 2b -C(O)OR 2b ;

[0525] L 2b is absent or optionally substituted C 1-10 alkylene;

[0526] R 2b , R' 2b and R” 2b Each instance of is independently H or C 1-10 alkyl; and

[0527] R z1 It is C 1-14 Alkyl or C 1-14 alkenyl, wherein the alkyl and alkenyl groups are optionally substituted.

[0528] In one embodiment, the steroid compound is a compound of formula (I):

[0529] or a stereoisomer, a mixture of stereoisomers, or a pharmaceutically acceptable salt thereof, wherein:

[0530] Is a single bond or a double bond;

[0531] Y is -C(O)O-, -O-, -NH-, -SC(O)O-, -OC(O)NR 2a -、-NR 2a C(O)NR 2a -、-OC(O)S-、-OC(O)O-、-NR 2a C(O)O-, -OC(O)-, -SC(O)-, -C(O)S-, -NR 2a -、-C(O)NR 2a -、-NR 2a C(O)-、-NR 2a C(O)S-、-SC(O)NR 2a -, -C(O)-, -OC(S)-, -C(S)O-, -OC(S)NR 2a -、-NR 2a C(S)O- or -S(O) 0-2 -, and wherein the leftward connection is in the direction of V1;

[0532] Y1 is absent, -C(O)O-, -O-, -NH-, -SC(O)O-, -OC(O)NR 2a -、-NR 2a C(O)NR 2a -、-OC(O)S-、-OC(O)O-、-NR 2a C(O)O-, -OC(O)-, -SC(O)-, -C(O)S-, -NR 2a -、-C(O)NR 2a -、-NR 2a C(O)-、-NR 2a C(O)S-、-SC(O)NR 2a -, -C(O)-, -OC(S)-, -C(S)O-, -OC(S)NR 2a -、-NR 2a C(S)O- or -S(O) 0-2 -, and wherein the leftward connection is in the direction of V1;

[0533] R 2a Each instance of is independently H, C 1-10 Alkyl, C 3-14 cycloalkyl or 3 to 14 membered heterocyclyl, and wherein the alkyl, cycloalkyl and heterocyclyl are independently optionally replaced by C 1-10 Alkyl, L 2b -OR 2b , Lb-SR 2b 、L 2b -NR 2b R'2b 、L 2b N + R 2b R' 2b R” 2b 、L 2b -OC(O)R 2b or L 2b -C(O)OR 2b replace;

[0534] R 201 、R 202 、R 203 、R 204 Each example is independently H, OH, alkyl, alkoxy, -L 2b -NR 21 R 22 、-L 2b -N + R 21 R 22 R 23 、L 2b -OC(O)R 21 or L 2b -C(O)OR 21 ;

[0535] m is an integer from 1 to 12;

[0536] m1 is an integer from 0 to 12;

[0537] V1 is NR 21 R 22 、N + R 21 R 22 R 23 , imidazole, pyridine, pyrrole, pyrrolidine, pyrazole, isothiazole, isoxazole, oxadiazine, oxazoline, dithioureaazole, isotriazole, tetrazole, pentazole, indole, indoline, isoxazole, pyrimidine, pyrazine, quinazoline, piperazine, piperidine, morpholine, pyran, quinoxaline, quinoline, quinolinone, isoquinoline, amidine, guanidine or urea;

[0538] M is absent or is one or more anions independently selected from chloride, bromide, iodide, sulfate, nitrate, perchlorate, formate, citrate, sulfonate, methanesulfonate, trifluoromethanesulfonate, trifluoroacetate, pyrosulfate, bisulfate, sulfite, bisulfite, phosphate, pyrophosphate, monohydrogen phosphate, dihydrogen phosphate, acetate, propionate, decanoate, octanoate, acrylate, isobutyrate, hexanoate, heptanoate, oxalate, malonate, succinate, fumarate, maleate, benzoate, phthalate, phenylacetate, lactate, glycolate, tartrate, mandelate, methanesulfonate, gluconate, aspartate, or glutamate;

[0539] R 21 、R 22 、R 23 Each instance of is independently H, optionally substituted C 1-8 Alkyl, -L 2b -OR 2b 、-L 2b -SR 2b 、-L 2b -NR 2b R' 2b R” 2b 、L 2b -OC(O)R 2b or L 2b -C(O)OR 2b ;

[0540] L 2b is absent or optionally substituted C 1-10 alkylene;

[0541] R 2b , R' 2b and R” 2b Each instance of is independently H or C 1-10 alkyl;

[0542] R z1 It is C 1-14 Alkyl or C 1-14 alkenyl, wherein the alkyl and alkenyl groups are optionally substituted; and

[0543] Provided that when Y is -NHC(O)-, at least one of the following conditions is met:

[0544] (i) m is an integer from 5 to 12; or

[0545] (ii) V1 is not N(CH3)2.

[0546] In one embodiment, the steroid compound is a compound of formula (IP):

[0547] or a stereoisomer, a mixture of stereoisomers, or a pharmaceutically acceptable salt thereof, wherein:

[0548] Is a single bond or a double bond;

[0549] Y1 does not exist, -C(O)O-, -O-, -NH-, -SC(O)O-, -OC(O)NR 2a -、-NR 2a C(O)NR 2a -、-OC(O)S-、-OC(O)O-、-NR 2a C(O)O-, -OC(O)-, -SC(O)-, -C(O)S-,-NR 2a -,-C(O)NR 2a -,-NR 2a C(O)-,-NR 2a C(O)S-,-SC(O)NR 2a -,-C(O)-,-OC(S)-, -C(S)O-, -OC(S)NR 2a -、-NR 2a C(S)O-, -S(O) 0-2 -or -OPO3-, and wherein the leftward connection is in the direction of V1;

[0550] R 2a Each instance of is independently H, C 1-10 Alkyl, C 3-14 cycloalkyl or 3 to 14 membered heterocyclyl, and wherein the alkyl, cycloalkyl and heterocyclyl are independently optionally replaced by C 1-10 Alkyl, L 2b -OR 2b 、L b -SR 2b 、L 2b -NR 2b R' 2b 、L 2b N + R 2b R' 2b R” 2b 、L 2b -OC(O)R 2b or L 2b -C(O)OR 2b replace;

[0551] R 201 、R 202、 R203 、R 204 Each example is independently H, OH, alkyl, alkoxy, -L 2b -NR 21 R 22 、-L 2b -N + R 21 R 22 R 23 、L 2b -OC(O)R 21 or L 2b -C(O)OR 21 ;

[0552] R 205 Absence, cations, H, C 1-6 Alkyl, C 1-6 Halogenated alkyl, C 2-6 Alkenyl or C 2-6 alkynyl, and wherein alkyl, haloalkyl, alkenyl and alkynyl are optionally substituted;

[0553] m is an integer from 1 to 12;

[0554] m1 is an integer from 0 to 12;

[0555] V1 is NR 21 R 22 、N + R 21 R 22 R 23 , imidazole, pyridine, pyrrole, pyrrolidine, pyrazole, (iso)thiazole, (iso)oxazole, oxadiazine, oxazoline, dithioureaazole, (iso)triazole, tetrazole, pentazole, indole, indoline, pyrimidine, pyrazine, quinazoline, piperazine, piperidine, morpholine, pyran, quinoxaline, quinoline, quinolinone, (iso)quinoline, amidine, guanidine or urea;

[0556] M is absent or is one or more anions independently selected from chloride, bromide, iodide, sulfate, nitrate, perchlorate, formate, citrate, sulfonate, methanesulfonate, trifluoromethanesulfonate, trifluoroacetate, pyrosulfate, bisulfate, sulfite, bisulfite, phosphate, pyrophosphate, monohydrogen phosphate, dihydrogen phosphate, acetate, propionate, decanoate, octanoate, acrylate, isobutyrate, hexanoate, heptanoate, oxalate, malonate, succinate, fumarate, maleate, benzoate, phthalate, phenylacetate, lactate, glycolate, tartrate, mandelate, methanesulfonate, gluconate, aspartate, or glutamate;

[0557] R 21 、R 22 、R23 Each instance of is independently H, optionally substituted C 1-8 Alkyl, -L 2b -OR 2b 、-L 2b -SR 2b 、-L 2b -NR 2b R' 2b R” 2b 、L 2b -OC(O)R 2b or L 2b -C(O)OR 2b ;

[0558] L 2b is absent or optionally substituted C 1-10 alkylene;

[0559] R 2b , R' 2b and R” 2b Each instance of is independently H or C 1-10 alkyl;

[0560] R z1 It is C 1-14 Alkyl or C 1-14 wherein said alkyl and alkenyl groups are optionally substituted. In one embodiment of formula (I), V1 is not NH2 or NH3 + In one embodiment of formula (I), when Y is -NR 2a C(O)-, V1 is not NH2 or NH3 + In one embodiment of formula (I), when Y is -NHC(O), V1 is not NH2 or NH3 + .

[0561] With respect to Formula (I) and Formula (IP), the following embodiments apply.

[0562] In one embodiment, Y is -C(O)O-, -OC(O)-, -O-, -NHC(O)O-, -OC(O)NH-, or -OC(O)O-. In one embodiment, Y is -C(O)O-. In one embodiment, Y is -O-. In one embodiment, Y is -NH-. In one embodiment, Y is -SC(O)O-. In one embodiment, Y is -OC(O)NR 2a In one embodiment, Y is -NR 2a C(O)NR 2aIn one embodiment, Y is -OC(O)S-. In one embodiment, Y is -OC(O)O-. In one embodiment, Y is -NR 2a In one embodiment, Y is -OC(O)-. In one embodiment, Y is -SC(O)-. In one embodiment, Y is -C(O)S-. In one embodiment, Y is -NR 2a In one embodiment, Y is -C(O)NR 2a In one embodiment, Y is -NR 2a C(O)-. In one embodiment, Y is -NR 2a C(O)S-. In one embodiment, Y is -SC(O)NR 2a -. In one embodiment, Y is -C(O)-. In one embodiment, Y is -OC(S)-. In one embodiment, Y is -C(S)O-. In one embodiment, Y is -OC(S)NR 2a In one embodiment, Y is -NR 2a C(S)O-. In one embodiment, Y is -S(O) 0-2 (eg, -S-). In these embodiments, the leftward linkage is in the direction of V1.

[0563] In one embodiment, Y1 is absent, -C(O)O-, -OC(O)-, -O-, -S-, SO, S(O)2, -O(CO)O-, -NHC(O)-, or -C(O)NH-. In one embodiment, Y1 is absent. In one embodiment, Y1 is -SC(O)O-. In one embodiment, Y1 is -OC(O)NR 2a -. In one embodiment, Y1 is -NR 2a C(O)NR 2a In one embodiment, Y1 is -OC(O)S-. In one embodiment, Y1 is -OC(O)O-. In one embodiment, Y1 is -NR 2a C(O)O-. In one embodiment, Y1 is -NR 2a C(O)S-. In one embodiment, Y1 is -SC(O)NR 2a -. In one embodiment, Y1 is -OC(S)NR 2a -. In one embodiment, Y1 is -NR 2aC(S)O-. In one embodiment, Y1 is -NHC(O)-. In one embodiment, Y1 is -C(O)NH-. In one embodiment, Y1 is -O(CO)O-. In one embodiment, Y1 is -O-. In these embodiments, the leftward connection is in the direction of V1.

[0564] In one embodiment, the steroid compound is a compound of Formula (I-A1), (I-A2), (I-A3), (I-A4), or (I-A5):

[0565] or a stereoisomer, a mixture of stereoisomers or a pharmaceutically acceptable salt thereof,

[0566] In one embodiment, the steroid compound is a compound of Formula (I-B1), (I-B2), (I-B3), (I-B4), (I-B5), (I-B6), (I-B7), or (I-B8):

[0567] or a stereoisomer, a mixture of stereoisomers, or a pharmaceutically acceptable salt thereof, wherein m3 is an integer from 1 to 11.

[0568] In one embodiment, the steroid compound is a compound of formula (I-P1):

[0569] or a stereoisomer, a mixture of stereoisomers, or a pharmaceutically acceptable salt thereof, and wherein m3 is an integer from 1 to 11.

[0570] In one embodiment, m is an integer from 1 to 12. In one embodiment, m is an integer from 5 to 12. In one embodiment, m is 5. In one embodiment, m is 6. In one embodiment, m is 7. In one embodiment, m is 8. In one embodiment, m is 9. In one embodiment, m is 10. In one embodiment, m is 11. In one embodiment, m is 12.

[0571] In one embodiment, m1 is an integer from 0 to 12. In one embodiment, m1 is an integer from 0 to 6. In one embodiment, m1 is 0, 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, or 12.

[0572] In one embodiment, m3 is an integer from 1 to 11. In one embodiment, m3 is an integer from 1 to 6. In one embodiment, m3 is 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, or 11.

[0573] In one embodiment, R2a It’s H.

[0574] In one embodiment, R 2a It is C 1-10 In one embodiment, R 2a It is C 1-6 In one embodiment, R 2a It is C 1-3 In one embodiment, R 2a In one embodiment, R 2a The alkyl group in 2b -OR 2b In one embodiment, R 2a The alkyl group in is unsubstituted. In one embodiment, R 2a The alkyl group in Lb-SR 2b In one embodiment, R 2a The alkyl group in 2b -NR 2b R' 2b In one embodiment, R 2a The alkyl group in 2b N + R 2b R' 2b R” 2b replace.

[0575] In one embodiment, R 2a It is C 3-14 In one embodiment, R 2a In one embodiment, R 2a is a C3-6 cycloalkyl group. 2a is C3, C4, C5 or C6 cycloalkyl. In one embodiment, R 2a The cycloalkyl group in is unsubstituted. In one embodiment, R 2a The cycloalkyl group in 1-10 In one embodiment, R 2a The cycloalkyl group in 2b -OR 2b In one embodiment, R 2a The cycloalkyl group in the Lb-SR 2b In one embodiment, R 2a The cycloalkyl group in 2b -NR 2b R' 2b In one embodiment, R 2a The cycloalkyl group in 2b N+ R 2b R' 2b R” 2b replace.

[0576] In one embodiment, R 2a is a 3 to 14 membered heterocyclyl. In one embodiment, R 2a is a 3 to 8 membered heterocyclyl. 2a is a 3- to 6-membered heterocyclic group. 2a is a 3 to 6-membered nitrogen-containing heterocyclic group. 2a is a 3 to 6-membered oxygen-containing heterocyclic group. 2a The heterocyclic group in is unsubstituted. In one embodiment, R 2a The heterocyclic group in 1-10 In one embodiment, R 2a The heterocyclic group in L 2b -OR 2b In one embodiment, R 2a The heterocyclic group in L b -SR 2b In one embodiment, R 2a The heterocyclic group in L 2b -NR 2b R' 2b In one embodiment, R 2a The heterocyclic group in L 2b N + R 2b R' 2b R” 2b replace.

[0577] In one embodiment, R 201 、R 202 、R 203 and R 204 Each instance of is independently H, C 1-6 Alkyl, -NR 21 R 22 、-N + R 21 R 22 R 23 、-(CH2) 0-6 -OC(O)R 21 or -(CH2) 0-6 -C(O)OR 21 .

[0578] In one embodiment, R 201 、R 202 、R203 and R 204 Each instance of is independently H, NH2, NH3 + , -CH2OC(O)CH3, -CH2C(O)OCH3, -OC(O)CH3 or -C(O)OCH3.

[0579] In one embodiment, V1 is NR 21 R 22 In one embodiment, V1 is N + R 21 R 22 R 23 .

[0580] In one embodiment, R 21 、R 22 、R 23 are independently H, hydroxyl, C 1-3 Alkyl or C 1-3 Hydroxyalkyl.

[0581] In one embodiment, V1 is an optionally substituted nitrogen-containing heterocyclic group. In one embodiment, V1 is an optionally substituted 3 to 8-membered nitrogen-containing heterocyclic group. In one embodiment, V1 is an optionally substituted nitrogen-containing heteroaryl group. In one embodiment, V1 is an optionally substituted 5 or 6-membered nitrogen-containing heteroaryl group. In one embodiment, V1 is selected from the group consisting of imidazole, pyridine, pyrrole, pyrrolidine, pyrazole, isothiazole, isoxazole, oxadiazine, oxazoline, dithiourea, isotriazole, tetrazole, pentazole, indole, indoline, isoxazole, pyrimidine, pyrazine, quinazoline, piperazine, piperidine, morpholine, pyran, quinoxaline, quinoline, quinolinone and isoquinoline.

[0582] In one embodiment, V1 is a nitrogen-containing non-heterocyclic group. In one embodiment, V1 is an optionally substituted amidine group. In one embodiment, V1 is an optionally substituted guanidine group. In one embodiment, V1 is an optionally substituted urea group.

[0583] In one embodiment, V1 is N(CH3)2, N + (CH3)3、N + CH3(CH2CH2OH)2、N + (CH3)2(CH2CH2OH), NHC(=N + H2)NH2, NHC(O)NH2, CH2C(=NH)NH2, N(CH3)(CH2CH2CO2CH3), N + (CH3)2(CH2CH2CO2CH3), N(CH3)(CH2CO2CH3) or N +(CH3)2(CH2CO2CH3). In one embodiment, V1 is N + CH3(CH2CH2OH)2、N + (CH3)2(CH2CH2OH) or N + (CH3)3.

[0584] In one embodiment, V1 is N(CH3)2. In one embodiment, V1 is N + (CH3)3. In one embodiment, V1 is N + CH3(CH2CH2OH)2. In one embodiment, V1 is N + (CH3)2(CH2CH2OH). In one embodiment, V1 is NHC(=N + In one embodiment, V1 is NHC(O)NH2. In one embodiment, V1 is CH2C(=NH)NH2.

[0585] In one embodiment, V1 is NH2. In one embodiment, V1 is NH3 + .

[0586] In one embodiment, R 21 、R 22 、R 23 Each independently is H, C 1-8 Alkyl, -L 2b -OR 2b 、-L 2b -SR 2b or-L2c-NR 2b R' 2b R” 2b In one embodiment, R 21 、R 22 、R 23 Each independently is H, C 1-6 Alkyl, -OR 2b 、-SR 2b or -NR 2b R' 2b R” 2b In one embodiment, R 21 、R 22 、R 23 are independently H, hydroxyl, C 1-3 Alkyl or C 1-3 Hydroxyalkyl.

[0587] In one embodiment, R 21 is H. In one embodiment, R 21 It is C 1-8In one embodiment, R 21 It is C 1-6 In one embodiment, R 21 It is C 1-3 In one embodiment, R 21 In one embodiment, R 21 In one embodiment, R 21 is propyl or isopropyl. In one embodiment, R 21 The alkyl group in is unsubstituted. In one embodiment, R 21 The alkyl group in is substituted by one or more halogens (i.e., F, Cl, Br, or I). 21 The alkyl group in the 2b In one embodiment, R 21 The alkyl group in Lb-SR 2b In one embodiment, R 21 The alkyl group in 2b -NR 2b R' 2b In one embodiment, R 21 The alkyl group in 2b N + R 2b R' 2b R” 2b replace.

[0588] In one embodiment, R 22 Yes-L 2b -OR 2b In one embodiment, R 22 Yes-L 2b -SR 2b In one embodiment, R 22 Yes-L2c-NR 2b R' 2b R” 2b .

[0589] In one embodiment, R 22 is H. In one embodiment, R 22 It is C 1-8 In one embodiment, R 22 It is C 1-6 In one embodiment, R 22 It is C 1-3 In one embodiment, R 22 In one embodiment, R 22 In one embodiment, R 22is propyl or isopropyl. In one embodiment, R 22 The alkyl group in is unsubstituted. In one embodiment, R 22 The alkyl group in is substituted by one or more halogens (i.e., F, Cl, Br, or I). 22 The alkyl group in the 2b In one embodiment, R 22 The alkyl group in Lb-SR 2b In one embodiment, R 22 The alkyl group in 2b -NR 2b R' 2b In one embodiment, R 22 The alkyl group in 2b N + R 2b R' 2b R” 2b replace.

[0590] In one embodiment, R 22 Yes-L 2b -OR 2b In one embodiment, R 22 Yes-L 2b -SR 2b In one embodiment, R 22 Yes-L2c-NR 2b R' 2b R” 2b .

[0591] In one embodiment, R 23 is H. In one embodiment, R 23 It is C 1-8 In one embodiment, R 23 It is C 1-6 In one embodiment, R 23 It is C 1-3 In one embodiment, R 23 In one embodiment, R 23 In one embodiment, R 23 is propyl or isopropyl. In one embodiment, R 23 The alkyl group in is unsubstituted. In one embodiment, R 23 The alkyl group in is substituted by one or more halogens (i.e., F, Cl, Br, or I). 23 The alkyl group in the 2b In one embodiment, R 23The alkyl group in Lb-SR 2b In one embodiment, R 23 The alkyl group in 2b -NR 2b R' 2b In one embodiment, R 23 The alkyl group in 2b N + R 2b R' 2b R” 2b replace.

[0592] In one embodiment, R 23 Yes-L 2b -OR 2b In one embodiment, R 23 Yes-L 2b -SR 2b In one embodiment, R 23 Yes-L2c-NR 2b R' 2b R” 2b .

[0593] In one embodiment, L 2b In one embodiment, L 2b is optionally substituted C 1-10 In one embodiment, L 2b is optionally substituted C 1-6 In one embodiment, L 2b is optionally substituted C 1-3 In one embodiment, L 2b The alkylene group in is unsubstituted. In one embodiment, L 2b The alkylene group is replaced by one or more halogen, hydroxyl or C 1-3 Alkoxy substitution.

[0594] In one embodiment, R 2b is H. In one embodiment, R 2b It is C 1-10 In one embodiment, R 2b It is C 1-6 In one embodiment, R 2b It is C 1-3 In one embodiment, R 2b In one embodiment, R 2b In one embodiment, R 2b is propyl or isopropyl. In one embodiment, R 2bThe alkyl group in is unsubstituted. In one embodiment, R 2b The alkyl group is replaced by one or more halogen, hydroxyl or C 1-3 Alkoxy substitution.

[0595] In one embodiment, R' 2b Is H. In one embodiment, R' 2b It is C 1-10 In one embodiment, R' 2b It is C 1-6 In one embodiment, R' 2b It is C 1-3 In one embodiment, R' 2b In one embodiment, R' 2b In one embodiment, R' 2b Is propyl or isopropyl. In one embodiment, R' 2b In one embodiment, the alkyl group in R' is unsubstituted. 2b The alkyl group is replaced by one or more halogen, hydroxyl or C 1-3 Alkoxy substitution.

[0596] In one embodiment, R" 2b is H. In one embodiment, R" 2b It is C 1-10 In one embodiment, R" 2b It is C 1-6 In one embodiment, R" 2b It is C 1-3 In one embodiment, R" 2b In one embodiment, R" 2b In one embodiment, R" 2b is propyl or isopropyl. In one embodiment, R" 2b The alkyl group in is unsubstituted. In one embodiment, R" 2b The alkyl group is replaced by one or more halogen, hydroxyl or C 1-3 Alkoxy substitution.

[0597] In one embodiment, M is one or more anions. In one embodiment, M is one or more pharmaceutically acceptable anions. In one embodiment, M is absent or is one or more anions independently selected from chloride, bromide, iodide, sulfate, nitrate, perchlorate, formate, citrate, sulfonate, methanesulfonate, trifluoromethanesulfonate, trifluoroacetate, pyrosulfate, bisulfate, sulfite, bisulfite, phosphate, pyrophosphate, monohydrogen phosphate, dihydrogen phosphate, acetate, propionate, decanoate, octanoate, acrylate, isobutyrate, hexanoate, heptanoate, oxalate, malonate, succinate, fumarate, maleate, benzoate, phthalate, phenylacetate, lactate, glycolate, tartrate, mandelate, methanesulfonate, gluconate, aspartate, or glutamate.

[0598] In one embodiment, M is chloride, bromide, iodide, sulfate, nitrate, perchlorate, formate, phosphate, pyrophosphate, citrate, sulfonate, methanesulfonate, trifluoromethanesulfonate or trifluoroacetate. In one embodiment, M is absent or is one or more of bromide, chloride or trifluoroacetate.

[0599] In one embodiment, R z1 It is C 1-14 Alkyl or C 1-14 Alkenyl, wherein the alkyl and alkenyl groups are optionally substituted with one or more hydroxy, oxo, nitro, cyano, halogen, C 1-6 Alkoxy, C2-C6 alkenyl, C2-C6 alkynyl, C3-C8 cycloalkyl, C6-C 10 In one embodiment, R z1 is C3-12 alkyl or C3-12 alkenyl. In one embodiment, R z1 It is C 7-10 Alkyl or C 7-10 In one embodiment, R z1 is C7 alkyl or C7 alkenyl. In one embodiment, R z1 is C8 alkyl or C8 alkenyl. In one embodiment, R z1 is C9 alkyl or C9 alkenyl. In one embodiment, R z1 It is C 10 Alkyl or C 10 Alkenyl.

[0600] In one embodiment, R z1 It is C 6-12 In one embodiment, R z1 is unsubstituted C6-12 In one embodiment, R z1 is substituted by one or more oxo or hydroxy groups. 6-12 alkyl.

[0601] In one embodiment, R z1 It is C 6-12 In one embodiment, R z1 is unsubstituted C 6-12 In one embodiment, R z1 is substituted by one or more oxo or hydroxy groups. 6-12 Alkenyl.

[0602] In one embodiment, R z1 yes In one embodiment, R z1 yes In one embodiment, R z1 yes In one embodiment, R z1 yes In one embodiment, R z1 yes In one embodiment, R z1 yes In one embodiment, R z1 yes

[0603] In one embodiment, the steroid compound is selected from:

[0604] or a stereoisomer, a mixture of stereoisomers or a pharmaceutically acceptable salt thereof.

[0605] In one embodiment, the steroid compound is not:

[0606] or a pharmaceutically acceptable salt thereof.

[0607] In one embodiment, provided herein is a steroid compound of formula (II-A):

[0608] or a stereoisomer, a mixture of stereoisomers, or a pharmaceutically acceptable salt thereof, wherein:

[0609] Each are independently a single bond or a double bond;

[0610] Y is -C(O)O-, -O-, -NH-, -SC(O)O-, -OC(O)NR 2a -、-NR 2a C(O)NR 2a -、-OC(O)S-、-OC(O)O-、-NR 2a C(O)O-, -OC(O)-, -SC(O)-, -C(O)S-, -NR 2a -、-C(O)NR 2a -、-NR 2a C(O)-、-NR 2a C(O)S-、-SC(O)NR 2a -, -C(O)-, -OC(S)-, -C(S)O-, -OC(S)NR 2a -、-NR 2a C(S)O- or -S(O) 0-2 -, and wherein the leftward connection is in the direction of V1;

[0611] Y1 is absent, -C(O)O-, -O-, -NH-, -SC(O)O-, -OC(O)NR 2a -、-NR 2a C(O)NR 2a -、-OC(O)S-、-OC(O)O-、-NR 2a C(O)O-, -OC(O)-, -SC(O)-, -C(O)S-, -NR 2a -、-C(O)NR 2a -、-NR 2a C(O)-、-NR 2a C(O)S-、-SC(O)NR 2a -, -C(O)-, -OC(S)-, -C(S)O-, -OC(S)NR 2a -、-NR 2a C(S)O- or -S(O) 0-2 -, and the leftward connection is in the direction of V1;

[0612] R 2a Each instance of is independently H, C 1-10 Alkyl, C 3-14 cycloalkyl or 3 to 14 membered heterocyclyl, and wherein the alkyl, cycloalkyl and heterocyclyl are independently optionally replaced by C 1-10 Alkyl, L 2b -OR 2b , Lb-SR 2b 、L 2b-NR 2b R' 2b 、L 2b N + R 2b R' 2b R” 2b 、L 2b -OC(O)R 2b or L 2b -C(O)OR 2b replace;

[0613] R 201 、R 202 、R 203 、R 204 Each example is independently H, OH, alkyl, alkoxy, -L 2b -NR 21 R 22 or -L 2b -N + R 21 R 22 R 23 、L 2b -OC(O)R 21 or L 2b -C(O)OR 21 ;

[0614] M is an integer from 3 to 12;

[0615] m1 is an integer from 0 to 12;

[0616] V1 is NR 21 、N + R 21 R 22 R 23 , imidazole, pyridine, pyrrole, pyrrolidine, pyrazole, (iso)thiazole, (iso)oxazole, oxadiazine, oxazoline, dithioureaazole, (iso)triazole, tetrazole, pentazole, indole, indoline, pyrimidine, pyrazine, quinazoline, piperazine, piperidine, morpholine, pyran, quinoxaline, quinoline, quinolinone, (iso)quinoline, amidine, guanidine or urea;

[0617] M is absent or is one or more anions independently selected from chloride, bromide, iodide, sulfate, nitrate, perchlorate, formate, citrate, sulfonate, methanesulfonate, trifluoromethanesulfonate, trifluoroacetate, pyrosulfate, bisulfate, sulfite, bisulfite, phosphate, pyrophosphate, monohydrogen phosphate, dihydrogen phosphate, acetate, propionate, decanoate, octanoate, acrylate, isobutyrate, hexanoate, heptanoate, oxalate, malonate, succinate, fumarate, maleate, benzoate, phthalate, phenylacetate, lactate, glycolate, tartrate, mandelate, methanesulfonate, gluconate, aspartate, or glutamate;

[0618] R 21 、R 22 、R 23 are each independently H, optionally substituted C 1-8 Alkyl, -L 2b -OR 2b 、-L 2b -SR 2b 、-L 2b -NR 2b R' 2b R” 2b 、L 2b -OC(O)R 2b or L 2b -C(O)OR 2b ;

[0619] L 2b is absent or optionally substituted C 1-10 alkylene;

[0620] R 2b , R' 2b and R” 2b Each of is independently H or C 1-10 alkyl; and

[0621] R z1 It is C 1-14 Alkyl or C 1-14 alkenyl, wherein the alkyl and alkenyl groups are optionally substituted; and provided that is not:

[0622] In one embodiment of formula (II-A), V1 is not NH2 or NH3 + In one embodiment of formula (II-A), when Y is -NR 2a C(O)-, V1 is not NH2 or NH3 +In one embodiment of formula (II-A), when Y is -NHC(O), V1 is not NH2 or NH3 + .

[0623] In one embodiment, the steroid compound is of formula (II-B):

[0624] or a stereoisomer, a mixture of stereoisomers, or a pharmaceutically acceptable salt thereof, wherein:

[0625] Y is -C(O)O-, -O-, -NH-, -SC(O)O-, -OC(O)NR 2a -、-NR 2a C(O)NR 2a -、-OC(O)S-、-OC(O)O-、-NR 2a C(O)O-, -OC(O)-, -SC(O)-, -C(O)S-, -NR 2a -、-C(O)NR 2a -、-NR 2a C(O)-、-NR 2a C(O)S-、-SC(O)NR 2a -, -C(O)-, -OC(S)-, -C(S)O-, -OC(S)NR 2a -、-NR 2a C(S)O- or -S(O) 0-2 -, and wherein the leftward connection is in the direction of V1;

[0626] Y1 is absent, -C(O)O-, -O-, -NH-, -SC(O)O-, -OC(O)NR 2a -、-NR 2a C(O)NR 2a -、-OC(O)S-、-OC(O)O-、-NR 2a C(O)O-, -OC(O)-, -SC(O)-, -C(O)S-, -NR 2a -、-C(O)NR 2a -、-NR 2a C(O)-、-NR 2a C(O)S-、-SC(O)NR 2a -, -C(O)-, -OC(S)-, -C(S)O-, -OC(S)NR 2a -、-NR 2a C(S)O- or -S(O) 0-2 -, and wherein the leftward connection is in the direction of V1;

[0627] R2a Each instance of is independently H, C 1-10 Alkyl, C 3-14 cycloalkyl or 3 to 14 membered heterocyclyl, and wherein the alkyl, cycloalkyl and heterocyclyl are independently optionally replaced by C 1-10 Alkyl, L 2b -OR 2b 、L b -SR 2b 、L 2b -NR 2b R' 2b 、L 2b N + R 2b R' 2b R” 2b 、L 2b -OC(O)R 2b or L 2b -C(O)OR 2b replace;

[0628] R 201 、R 202、 R 203 、R 204 Each example is independently H, OH, alkyl, alkoxy, -L 2b -NR 21 R 22 or -L 2b -N + R 21 R 22 R 23 、L 2b -OC(O)R 21 or L 2b -C(O)OR 21 ;

[0629] m is an integer from 5 to 12;

[0630] m1 is an integer from 0 to 12;

[0631] V1 is NR 21 R 22 、N + R 21 R 22 R 23 , imidazole, pyridine, pyrrole, pyrrolidine, pyrazole, (iso)thiazole, (iso)oxazole, oxadiazine, oxazoline, dithioureaazole, (iso)triazole, tetrazole, pentazole, indole, indoline, pyrimidine, pyrazine, quinazoline, piperazine, piperidine, morpholine, pyran, quinoxaline, quinoline, quinolinone, (iso)quinoline, amidine, guanidine or urea;

[0632] M is absent or is one or more anions independently selected from chloride, bromide, iodide, sulfate, nitrate, perchlorate, formate, citrate, sulfonate, methanesulfonate, trifluoromethanesulfonate, trifluoroacetate, pyrosulfate, bisulfate, sulfite, bisulfite, phosphate, pyrophosphate, monohydrogen phosphate, dihydrogen phosphate, acetate, propionate, decanoate, octanoate, acrylate, isobutyrate, hexanoate, heptanoate, oxalate, malonate, succinate, fumarate, maleate, benzoate, phthalate, phenylacetate, lactate, glycolate, tartrate, mandelate, methanesulfonate, gluconate, aspartate, or glutamate;

[0633] R 21 、R 22 、R 23 are each independently H, optionally substituted C 1-8 Alkyl, -L 2b -OR 2b 、-L 2b -SR 2b 、-L 2b -NR 2b R' 2b R” 2b 、L 2b -OC(O)R 2b or L 2b -C(O)OR 2b ;

[0634] L 2b is absent or optionally substituted C 1-10 alkylene;

[0635] R 2b , R' 2b and R” 2b Each of is independently H or C 1-10 alkyl.

[0636] In one embodiment, Y1 in the steroid compound of formula (II-B) is -C(O)O-, -OC(O)-, -O-, -S-, -NR 2a -、-OC(O)NR 2a -or-NR 2a C(O)-. In one embodiment, Y1 in the steroid compound of formula (II-B) is -C(O)O-, -OC(O)-, -O-, -S-, -NH-, -NCH3-, -OC(O)NH-, or -NHC(O)-. In one embodiment, Y1 in the steroid compound of formula (II-B) is -O-. In one embodiment, Y1 in the steroid compound of formula (II-B) is -S-.

[0637] In another embodiment, the steroid compound is a compound of formula (II-C1):

[0638] or a stereoisomer, a mixture of stereoisomers, or a pharmaceutically acceptable salt thereof, wherein:

[0639] R z1 yes

[0640] R 2a is H, optionally substituted C 1-8 Alkyl, -L 2b -OR 2b 、L 2b -OC(O)R 2b or L 2b -C(O)OR 2b , preferably H, -CH3, -CH2C(O)OCH3 or -CH2C(O)OH;

[0641] V1 is NR 21 R 22 or N + R 21 R 22 R 23 ; where R 21 、R 22 、R 23 are each independently H, optionally substituted C 1-8 Alkyl, -L 2b -OR 2b 、-L 2b -SR 2b 、-L 2b -NR 2b R' 2b R” 2b 、L 2b -OC(O)R 2b or L 2b -C(O)OR 2b ; and R 21 and R 22 Not simultaneously hydrogen;

[0642] L 2b is absent or optionally substituted C 1-10 alkylene;

[0643] R 2b , R' 2b and R” 2b Each of is independently H or C 1-10 alkyl;

[0644] M is absent or is one or more anions independently selected from chloride, bromide, iodide, sulfate, nitrate, perchlorate, formate, citrate, sulfonate, methanesulfonate, trifluoromethanesulfonate, trifluoroacetate, pyrosulfate, bisulfate, sulfite, bisulfite, phosphate, pyrophosphate, monohydrogen phosphate, dihydrogen phosphate, acetate, propionate, decanoate, octanoate, acrylate, isobutyrate, hexanoate, heptanoate, oxalate, malonate, succinate, fumarate, maleate, benzoate, phthalate, phenylacetate, lactate, glycolate, tartrate, mandelate, methanesulfonate, gluconate, aspartate, or glutamate;

[0645] m is an integer from 1 to 12, for example, 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11 or 12.

[0646] In another embodiment, the steroid compound is a compound of formula (II-C2):

[0647] or a stereoisomer, a mixture of stereoisomers, or a pharmaceutically acceptable salt thereof, wherein:

[0648] R z1 yes

[0649] V1 is NR 21 R 22 or N + R 21 R 22 R 23 ; where R 21 、R 22 、R 23 are each independently H, optionally substituted C 1-8 Alkyl, -L 2b -OR 2b 、-L 2b -SR 2b 、-L 2b -NR 2b R' 2b R” 2b 、L 2b -OC(O)R 2b or L 2b -C(O)OR 2b ; and R 21 and R 22 Not simultaneously hydrogen;

[0650] L 2b is absent or optionally substituted C 1-10 alkylene;

[0651] R 2b , R' 2b and R” 2b Each of is independently H or C 1-10 alkyl;

[0652] M is absent or is one or more anions independently selected from chloride, bromide, iodide, sulfate, nitrate, perchlorate, formate, citrate, sulfonate, methanesulfonate, trifluoromethanesulfonate, trifluoroacetate, pyrosulfate, bisulfate, sulfite, bisulfite, phosphate, pyrophosphate, monohydrogen phosphate, dihydrogen phosphate, acetate, propionate, decanoate, octanoate, acrylate, isobutyrate, hexanoate, heptanoate, oxalate, malonate, succinate, fumarate, maleate, benzoate, phthalate, phenylacetate, lactate, glycolate, tartrate, mandelate, methanesulfonate, gluconate, aspartate, or glutamate;

[0653] m is an integer from 1 to 12, for example, 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11 or 12.

[0654] In one embodiment, the steroid compound is not: or a pharmaceutically acceptable salt thereof.

[0655] In one embodiment, provided herein are the following compounds:

[0656] or a stereoisomer, a mixture of stereoisomers or a pharmaceutically acceptable salt thereof.

[0657] In one embodiment, the molar percentage of the steroid compound to the total lipids is 3-15 mol%, preferably 5-10 mol%, for example 3 mol%, 4 mol%, 5 mol%, 6 mol%, 7 mol%, 8 mol%, 9 mol%, 10 mol%, 11 mol%, 12 mol%, 13 mol%, 14 mol% or 15 mol%.

[0658] 5.2.2. Ionizable lipids

[0659] In one embodiment, the lipid nanoparticle further comprises an ionizable lipid. In one embodiment, the ionizable lipid is different from the steroid compound described in Section 0. In one embodiment, the lipid nanoparticle comprises an ionizable lipid and a steroid compound having a cationic group.

[0660] In one embodiment, the pKa of the ionizable lipid is from about 4 to about 14. In one embodiment, the pKa of the ionizable lipid is from about 5 to about 7. In one embodiment, the pKa of the ionizable lipid is from about 6 to about 7. In one embodiment, the pKa of the ionizable lipid is greater than 7. In one embodiment, the pKa of the ionizable lipid is about 4, about 5, about 6, about 7, about 8, about 9, about 10, or about 11. In one embodiment, the pKa of the ionizable lipid is 5.0, 5.1, 5.2, 5.3, 5.4, 5.5, 5.6, 5.7, 5.8, 5.9, 6.0, 6.1, 6.2, 6.3, 6.4, 6.5, 7.0, 7.5, 8.0, 8.5, 9.0, 9.5, 10.0, 10.5, 11.0, or greater than 11. In one embodiment, the ionizable lipid becomes positively charged at physiological pH (i.e., pH 7.4).

[0661] In one embodiment, the ionizable lipid comprises one or more groups that are protonated at physiological pH, but can be deprotonated and uncharged at a pH greater than 8, greater than 9, or greater than 10. In one embodiment, the ionizable lipid comprises one or more tertiary amine groups. In one embodiment, the ionizable lipid comprises one, two, three, or four C6-C24 alkyl or alkenyl lipid groups. These lipid groups can be connected by functional groups (e.g., ester groups or amide groups), or can be further added to the sulfur atom by Michael addition.

[0662] In one embodiment, the ionizable lipid comprises at least one ionizable head group and at least one biodegradable hydrophobic chain, wherein the ionizable lipid has a logP of at least about 10.1 and a pKa of about 4 to about 11; and the biodegradable hydrophobic chain has the structural formula -R 16 -M0-R 17 ; where R 16 It is C 4-14 Alkylene, C 4-14 Alkenylene or C 4-14 Alkynylidene; R 17 It is C 6-20 and M0 is a biodegradable group, and wherein the alkylene, alkenylene, alkynylene and alkyl are optionally substituted. In one embodiment, the ionizable head group is a tertiary amine group. In one embodiment, the tertiary amine group of the ionizable lipid comprises a -CH2CH2OH moiety.

[0663] In one embodiment, the ionizable lipid comprises a tertiary amine group and two biodegradable hydrophobic chains, wherein each hydrophobic chain is independently -R 16 -C(O)OR 17 , and where R 16 In one embodiment, R 17 One of them is a forked C 6-20 alkyl, and another R 17 It is linear C 6-20 alkyl.

[0664] In one embodiment, M0 is an ester group, an amide group, a thioester group, a carbonate group, a carbamate group, a carbamoylthioester group, a urea group or an ether group. In one embodiment, M0 is an ester group.

[0665] In one embodiment, the ionizable lipid is a compound of formula (III):

[0666] or a stereoisomer, mixture of stereoisomers, tautomer, isotopomer or pharmaceutically acceptable salt thereof, wherein:

[0667] Each instance of R3 and R4 is independently C 1-10 Alkyl, C 2-10 Alkenyl, C 2-10 Alkynyl, C3-C 14 Cycloalkyl, 3 to 14 membered heterocyclic group, (C 1-10 Alkylene)-(C3-C 14 Cycloalkyl) or (C 1-10 alkylene)-(3 to 14 membered heterocyclyl), or R3 and R4 together with the nitrogen atom to which they are attached form a 3 to 14 membered heterocyclyl, or R4 and one R0' together with the atom to which they are attached form a 4 to 10 membered heterocyclyl or a 5 to 10 membered heteroaryl, and wherein said alkyl, alkenyl, alkynyl, cycloalkyl, heterocyclyl, heteroaryl and alkylene are independently optionally substituted with one or more R*;

[0668] Each instance of R0' is independently optionally substituted methylene, or wherein the two substituents of R0' together with the methylene to which they are attached form a 3- to 8-membered cycloalkylene, and wherein the cycloalkylene is optionally substituted with one or more R**;

[0669] k is 0, 1, 2, 3, 4, 5, or 6;

[0670] j is 0 or 1;

[0671] The dotted line connecting Q and W is either non-existent or a bond;

[0672] W is C, CH or N, provided that when W is N, j is 0 and the dotted bond connecting Q and W is absent;

[0673] G5 does not exist, C 1-24 Alkylene, C 2-24 Alkenylene, C3-C8 cycloalkylene, C3-C8 cycloalkenylene, wherein the alkylene, alkenylene, cycloalkylene, cycloalkenylene are optionally substituted with one or more R**;

[0674] G1, G2, G3 and G4 are each independently absent, C 1-13 Alkylene, C 2-13 Alkenylene or C 2-13 Alkyne, wherein the alkylene, alkenylene and alkynylene groups are optionally substituted by one or more R s replace;

[0675] The total length of G1 and G2 is 3, 4, 5, 6, 7, 8, 9, 10, 11, 12 or 13 carbon atoms;

[0676] The total length of G3 and G4 is 3, 4, 5, 6, 7, 8, 9, 10, 11, 12 or 13 carbon atoms;

[0677] R5, R6, R7 and R8 are each independently hydrogen or optionally substituted C 1-8 alkyl;

[0678] When the dashed line connecting Q and W is absent, Q is absent or selected from -C(O)O-, -O-, -NH-, -SC(O)O-, -OC(O)NR b -、-NR b C(O)NR b -、-OC(O)S-、-OC(O)O-、-NR b C(O)O-, -OC(O)-, -SC(O)-, -C(O)S-, -NRb-, -C(O)NR b -、-NR b C(O)-、-NR b C(O)S-、-SC(O)NR b -, -C(O)-, -OC(S)-, -C(S)O-, -OC(S)NR b -、-NR b C(S)O-, -SS-, or -S(O) 0-2 -, where the rightward connection is in the direction of W;

[0679] When the dashed line connecting Q and W is a chemical bond, G5 is absent, and Q and W form a 5- to 10-membered monocyclic or fused ring, optionally wherein the ring is substituted with one or more R**;

[0680] M1 and M2 are each independently absent, or -C(O)O-, -O-, -SC(O)O-, -OC(O)NR a -、-NR a C(O)NR a -、-OC(O)S-、-OC(O)O-、-NR a C(O)O-, -OC(O)-, -SC(O)-, -C(O)S-, -NR a -、-C(O)NR a -、-NR a C(O)-、-NR a C(O)S-、-SC(O)NR a -, -C(O)-, -OC(S)-, -C(S)O-, -OC(S)NR a -、-NR a C(S)O-, -SS-, or -S(O) 0-2 -;

[0681] Each instance of R0 is independently -(CRR')-;

[0682] Each instance of R and R' is independently H, C 1-20 Alkyl, L a -OR a 、-L a -SR a or -L a -NR a R' a , or R and R' together with the carbon to which they are attached form C 3-8 cycloalkylene;

[0683] Q3 and Q4 are each independently H, -(CRR')-, C 6-10 an aryl or steroid moiety;

[0684] Each instance of A1, A2, A3, A4 is independently -(CR 18 R 18 -CR 18 =CR 18 )-or-(CR 18 R 18 -C≡C)-;

[0685] R 18 Each example of C is independently 1-20 alkyl;

[0686] Each instance of N1 and N2 is independently a biodegradable group;

[0687] Z does not exist, C 1-10 Alkylene or -OP(O)(OH)-O-;

[0688] Each of the dashed lines between Z-Q3 or Z-Q4 is independently absent or a chemical bond, provided that when Z is absent, both dashed lines are absent;

[0689] m, n, q, r, u, v, y and z are each independently 0, 1, 2, 3, 4, 5, 6, 7, 8, 9 or 10;

[0690] o, p, w and x are each independently 0, 1 or 2;

[0691] s and t are each independently 0 or 1;

[0692] The number of atoms between G1 and Q3 or between G3 and Q4 is 8 to 30;

[0693] L a and Le independently do not exist or are C 1-20 alkylene;

[0694] L b and L f Each independently does not exist or is C 1-10 alkylene;

[0695] L c Does not exist or C 1-8 alkylene;

[0696] L d Does not exist or C 1-14 alkylene;

[0697] R* is hydroxyl, halogen, C 1-10 Alkyl, C 1-10 Halogenated alkyl, C 1-10 Hydroxyalkyl, -L b -OR b 、-L b -SR b or -L b -NR b R' b ;

[0698] R** is hydroxyl, halogen, C 1-8 Alkyl, C 1-8 Halogenated alkyl, C 1-8 Hydroxyalkyl, -L b -ORb 、-L b -SR b or -L b -NR b R' b ;

[0699] R s It is C 1-14 Alkyl, -L b -OR b 、-L b -SR b or -L b -NR b R' b ;

[0700] R a and R' a Each independently is H, C 1-20 Alkyl, C 3-14 Cycloalkyl or 3 to 14 membered heterocyclyl, wherein the alkyl, cycloalkyl and heterocyclyl are optionally substituted by one or more selected from hydroxy, halogen, C 1-20 Alkyl, -L e -OR e 、-L e -SR e or -L e -NR e R' e The group substitution;

[0701] R b and R' b Each independently is H, C 1-10 Alkyl, C 3-14 Cycloalkyl or 3 to 14 membered heterocyclyl, wherein the alkyl, cycloalkyl and heterocyclyl are optionally substituted by one or more selected from hydroxy, halogen, C 1-10 Alkyl, -L f -OR f 、-L f -SR f or -L f -NR f R' f The group substitution;

[0702] R c and R' c are each independently hydrogen or C 1-8 alkyl;

[0703] R d and R' d are each independently hydrogen or C 1-14 alkyl;

[0704] Re and R' e are each independently hydrogen or C 1-20 alkyl; and

[0705] R f and R' f are each independently hydrogen or C 1-10 alkyl.

[0706] In one embodiment, m is 0. In one embodiment, m is 1. In one embodiment, m is 2. In one embodiment, m is 3. In one embodiment, m is 4. In one embodiment, m is 5. In one embodiment, m is 6. In one embodiment, m is 7. In one embodiment, m is 8. In one embodiment, m is 9. In one embodiment, m is 10.

[0707] In one embodiment, n is 0. In one embodiment, n is 1. In one embodiment, n is 2. In one embodiment, n is 3. In one embodiment, n is 4. In one embodiment, n is 5. In one embodiment, n is 6. In one embodiment, n is 7. In one embodiment, n is 8. In one embodiment, n is 9. In one embodiment, n is 10.

[0708] In one embodiment, q is 0. In one embodiment, q is 1. In one embodiment, q is 2. In one embodiment, q is 3. In one embodiment, q is 4. In one embodiment, q is 5. In one embodiment, q is 6. In one embodiment, q is 7. In one embodiment, q is 8. In one embodiment, q is 9. In one embodiment, q is 10.

[0709] In one embodiment, r is 0. In one embodiment, r is 1. In one embodiment, r is 2. In one embodiment, r is 3. In one embodiment, r is 4. In one embodiment, r is 5. In one embodiment, r is 6. In one embodiment, r is 7. In one embodiment, r is 8. In one embodiment, r is 9. In one embodiment, r is 10.

[0710] In one embodiment, u is 0. In one embodiment, u is 1. In one embodiment, u is 2. In one embodiment, u is 3. In one embodiment, u is 4. In one embodiment, u is 5. In one embodiment, u is 6. In one embodiment, u is 7. In one embodiment, u is 8. In one embodiment, u is 9. In one embodiment, u is 10.

[0711] In one embodiment, v is 0. In one embodiment, v is 1. In one embodiment, v is 2. In one embodiment, v is 3. In one embodiment, v is 4. In one embodiment, v is 5. In one embodiment, v is 6. In one embodiment, v is 7. In one embodiment, v is 8. In one embodiment, v is 9. In one embodiment, v is 10.

[0712] In one embodiment, y is 0. In one embodiment, y is 1. In one embodiment, y is 2. In one embodiment, y is 3. In one embodiment, y is 4. In one embodiment, y is 5. In one embodiment, y is 6. In one embodiment, y is 7. In one embodiment, y is 8. In one embodiment, y is 9. In one embodiment, y is 10.

[0713] In one embodiment, z is 0. In one embodiment, z is 1. In one embodiment, z is 2. In one embodiment, z is 3. In one embodiment, z is 4. In one embodiment, z is 5. In one embodiment, z is 6. In one embodiment, z is 7. In one embodiment, z is 8. In one embodiment, z is 9. In one embodiment, z is 10.

[0714] In one embodiment, m+q+u+y is an integer from 3 to 25. In one embodiment, m+q+u+y is an integer from 5 to 20. In one embodiment, m+q+u+y is an integer from 5 to 20. In one embodiment, m+q+u+y is an integer from 5 to 20. In one embodiment, m+q+u+y is an integer from 5 to 18. In one embodiment, m+q+u+y is an integer from 6 to 12. In one embodiment, m+q+u+y is 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17 or 18.

[0715] In one embodiment, n+r+v+z is an integer from 3 to 25. In one embodiment, n+r+v+z is an integer from 5 to 20. In one embodiment, n+r+v+z is an integer from 5 to 20. In one embodiment, n+r+v+z is an integer from 5 to 20. In one embodiment, n+r+v+z is an integer from 5 to 18. In one embodiment, n+r+v+z is an integer from 6 to 12. In one embodiment, n+r+v+z is 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17 or 18.

[0716] In one embodiment, o is 0. In one embodiment, o is 1. In one embodiment, o is 2.

[0717] In one embodiment, p is 0. In one embodiment, p is 1. In one embodiment, p is 2.

[0718] In one embodiment, w is 0. In one embodiment, w is 1. In one embodiment, w is 2.

[0719] In one embodiment, x is 0. In one embodiment, x is 1. In one embodiment, x is 2.

[0720] In one embodiment, o+p is 0. In one embodiment, o+p is 1. In one embodiment, o+p is 2. In one embodiment, w+x is 0. In one embodiment, w+x is 1. In one embodiment, w+x is 2. In one embodiment, o+p+w+x is 0. In one embodiment, o+p+w+x is 1. In one embodiment, o+p+w+x is 2. In one embodiment, o+p+w+x is 3. In one embodiment, o+p+w+x is 4.

[0721] In one embodiment, R0 is CH2. In one embodiment, R0 is CH(C 1-16 In one embodiment, R0 is CH(C 1-14 In one embodiment, R0 is CH(C 1-12 In one embodiment, R0 is CH(C 1-6 In one embodiment, R0 is CHCH3. In one embodiment, R0 is C(CH3)2. In one embodiment, R0 is C(C 1-12 In one embodiment, R0 is CH(L a -OR a In one embodiment, R0 is CH(L a -SR a In one embodiment, R0 is CH(L a -NR a R' a In one embodiment, R0 is CH(OR a In one embodiment, R0 is CH(SR a In one embodiment, R0 is CH(NR a R' a In one embodiment, R0 is CHN(C 1-12In one embodiment, R0 is CH(CH2) 1-6 NR a R' a In one embodiment, R0 is CH(CH2) 1-6 N(C 1-12 Alkyl)2.

[0722] In one embodiment, Q3 is H. In one embodiment, Q3 is -(CRR')-. In one embodiment, Q3 is -(CH2)-. In one embodiment, Q4 is H. In one embodiment, Q4 is -(CRR')-. In one embodiment, Q4 is -(CH2)-.

[0723] In one embodiment, A1 is absent. In one embodiment, A1 is (CR 18 R 18 -CR 18 =CR 18 In one embodiment, A1 is -(CHR 18 -CH=CH)-. In one embodiment, A1 is -(CH2-CH=CH)-. In one embodiment, the C=C bond in A1 is in the form of a Z configuration. In one embodiment, the C=C bond in A1 is in the form of an E configuration. In one embodiment, A1 is -(CR 18 R 18 -C≡C)-. In one embodiment, A1 is -(CHR 18 In one embodiment, A1 is -(CH2-C≡C)-.

[0724] In one embodiment, A2 is absent. In one embodiment, A2 is (CR 18 R 18 -CR 18 =CR 18 In one embodiment, A2 is -(CHR 18 -CH=CH)-. In one embodiment, A2 is -(CH2-CH=CH)-. In one embodiment, the C=C bond in A2 is in the form of a Z configuration. In one embodiment, the C=C bond in A2 is in the form of an E configuration. In one embodiment, A2 is -(CR 18 R 18 -C≡C)-. In one embodiment, A2 is -(CHR18-C≡C)-. In one embodiment, A2 is -(CH2-C≡C)-.

[0725] In one embodiment, A3 is absent. In one embodiment, A3 is (CR18 R 18 -CR 18 =CR 18 In one embodiment, A3 is -(CHR 18 -CH=CH)-. In one embodiment, A3 is -(CH2-CH=CH)-. In one embodiment, the C=C bond in A3 is in the form of a Z configuration. In one embodiment, the C=C bond in A3 is in the form of an E configuration. In one embodiment, A3 is -(CR 18 R 18 -C≡C)-. In one embodiment, A3 is -(CHR 18 In one embodiment, A3 is -(CH2-C≡C)-.

[0726] In one embodiment, A4 is absent. In one embodiment, A4 is (CR 18 R 18 -CR 18 =CR 18 In one embodiment, A4 is -(CHR 18 -CH=CH)-. In one embodiment, A4 is -(CH2-CH=CH)-. In one embodiment, the C=C bond in A4 is in the form of a Z configuration. In one embodiment, the C=C bond in A4 is in the form of an E configuration. In one embodiment, A4 is -(CR 18 R 18 -C≡C)-. In one embodiment, A4 is -(CHR 18 In one embodiment, A4 is -(CH2-C≡C)-.

[0727] In one embodiment, R 18 is H. In one embodiment, R 18 It is C 1-20 In one embodiment, R 18 It is C 1-16 In one embodiment, R 18 It is C 1-12 In one embodiment, R 18 It is C 1-6 alkyl.

[0728] In one embodiment, s is 0. In one embodiment, s is 1. In one embodiment, t is 0. In one embodiment, t is 1.

[0729] In one embodiment, N1 is absent. In one embodiment, N1 is a biodegradable group. In one embodiment, N1 is an ester group. In one embodiment, N1 is an amide group. In one embodiment, N1 is a thioester group. In one embodiment, N1 is a carbonate group. In one embodiment, N1 is a carbamate group. In one embodiment, N1 is a carbamoylthioester group. In one embodiment, N1 is a urea group. In one embodiment, N1 is an imine group. In one embodiment, N1 is an oxime group. In one embodiment, N1 is a disulfide group.

[0730] In one embodiment, N2 is absent. In one embodiment, N2 is a biodegradable group. In one embodiment, N2 is an ester group. In one embodiment, N2 is an amide group. In one embodiment, N2 is a thioester group. In one embodiment, N2 is a carbonate group. In one embodiment, N2 is a carbamate group. In one embodiment, N2 is a carbamoylthioester group. In one embodiment, N2 is a urea group. In one embodiment, N2 is an imine group. In one embodiment, N2 is an oxime group. In one embodiment, N2 is a disulfide group.

[0731] In one embodiment, Z is absent. In one embodiment, Z is absent, and the dashed line between Q4 and Z and the dashed line between Q3 and Z are both absent. In one embodiment, Z is absent, and Q4 and Q3 are both H.

[0732] In one embodiment, Z is C 1-10 In one embodiment, Z is C 1-6 In one embodiment, Z is C 1-3 In one embodiment, Z is -OP(O)(OH)-O-.

[0733] In one embodiment, Z is absent and the number of carbon atoms between G1 and Q3 is from 8 to 30. In one embodiment, Z is absent and the number of carbon atoms between G1 and Q3 is from 10 to 25. In one embodiment, Z is absent and the number of carbon atoms between G1 and Q3 is from 10 to 20.

[0734] In one embodiment, Z is absent and the number of carbon atoms between G3 and Q4 is from 8 to 30. In one embodiment, Z is absent and the number of carbon atoms between G3 and Q4 is from 10 to 25. In one embodiment, Z is absent and the number of carbon atoms between G3 and Q4 is from 10 to 20.

[0735] In one embodiment, the ionizable lipid is a compound of formula (IV'):

[0736] or a stereoisomer, a mixture of stereoisomers or a pharmaceutically acceptable salt thereof, wherein

[0737] j is 0 or 1;

[0738] W is CH or N, provided that when W is N, j is 0;

[0739] k is 0, 1, 2, 3, 4, 5, 6, 7, or 8;

[0740] Each instance of R0' is independently optionally substituted methylene, or wherein the two substituents of R0' together with the methylene to which they are attached form a 3- to 8-membered cycloalkylene, and wherein the cycloalkylene is optionally substituted with one or more R**;

[0741] M1 and M2 are each independently -C(O)O-, -O-, -SC(O)O-, -OC(O)NR a -、-NR a C(O)NR a -、-OC(O)S-、-OC(O)O-、-NR a C(O)O-, -OC(O)-, -SC(O)-, -C(O)S-, -NR a -、-C(O)NR a -、-NR a C(O)-、-NR a C(O)S-、-SC(O)NR a -, -C(O)-, -OC(S)-, -C(S)O-, -OC(S)NR a -、-NR a C(S)O-, -SS-, or -S(O) 0-2 -;

[0742] Q does not exist, -C(O)O-, -O-, -NH-, -SC(O)O-, -OC(O)NRb-, -NRbC(O)NRb-, -OC(O)S-, -OC(O)O-, -NRbC(O)O-, -OC(O)-, -SC(O)-, -C(O )S-, -NRb-, -C(O)NRb-, -NRbC(O)-, -NRbC(O)S-, -SC(O)NRb-, -C(O)-, -OC(S)-, -C(S)O-, -OC(S)NRb-, -NRbC(S)O-, -SS- or -S(O) 0-2 -;

[0743] R a and Rb are each independently H, C 1-20 Alkyl, C 3-14 Cycloalkyl or 3 to 14 membered heterocyclyl, wherein the alkyl, cycloalkyl and heterocyclyl are optionally substituted by one or more selected from hydroxy, halogen, C 1-20 Alkyl, -Le-ORe, -Le-SRe or -Le-NReR'e;

[0744] G5 is absent or optionally substituted C 1-8 alkylene;

[0745] R1 and R2 are each independently C 4-20 Alkyl, C 4-20 Alkenyl or C 4-20 Alkynyl, wherein one or more methylene units in R1 and R2 are independently optionally replaced by -NH- or -N(C 1-20 alkyl), wherein the alkyl, alkenyl and alkynyl groups are optionally substituted;

[0746] Each instance of R3 and R4 is independently H, C 1-10 Alkyl, C 2-10 Alkenyl, C 2-10 Alkynyl, C3-C 14 cycloalkyl or 3 to 14 membered heterocyclyl, or R3 and R4 together with the nitrogen atom to which they are attached form a 3 to 14 membered heterocyclyl, or R4 and one R0' together with the atom to which they are attached form a 4 to 10 membered heterocyclyl; and wherein said alkyl, alkenyl, alkynyl, cycloalkyl, heterocyclyl is optionally substituted with one or more R*; and

[0747] R5, R6, R7 and R8 are each independently hydrogen or optionally substituted C 1-8 alkyl.

[0748] In one embodiment, the ionizable lipid is a compound of formula (IV'-A):

[0749] or a stereoisomer, a mixture of stereoisomers or a pharmaceutically acceptable salt thereof.

[0750] In one embodiment, the ionizable lipid is a compound of formula (VIII):

[0751] or a stereoisomer, a mixture of stereoisomers or a pharmaceutically acceptable salt thereof.

[0752] In one embodiment, the ionizable lipid is a compound of formula (V'), (VI') or (VII'):

[0753] or a stereoisomer, a mixture of stereoisomers or a pharmaceutically acceptable salt thereof, wherein

[0754] k is 0, 1, 2, 3, 4, or 5;

[0755] L1 and L2 are each independently -(CRR')2-, -CH=CH-, -C≡C- or -NR"-;

[0756] One of L3 and L5 is -(CR s R s ')2-, -CH=CH- or -C≡C-, and the other is absent;

[0757] One of L4 and L6 is -(CR s R s ')2-, -CH=CH- or -C≡C-, and the other is absent;

[0758] G 1a , G 1b , G 2a , G 2b , G 3a , G 3b , G 4a and G 4b Each of which is independently absent or optionally substituted C1-7 alkylene;

[0759] G 1a , G 1b , G 2a and G 2b The total length of is 1, 2, 3, 4, 5, 6 or 7 carbon atoms;

[0760] G 3a , G 3b , G 4a and G 4b The total length of is 1, 2, 3, 4, 5, 6 or 7 carbon atoms;

[0761] R5, R6, R7 and R8 are each independently hydrogen or optionally substituted C 1-6 alkyl;

[0762] G7, G8, G9 and G 10 Each independently absent or optionally substituted C 1-12 Alkylene, provided that the total length of G7 and G8 is 4, 5, 6, 7, 8, 9, 10, 11 or 12 carbon atoms, and G9 and G 10 has a total length of 4, 5, 6, 7, 8, 9, 10, 11 or 12 carbon atoms;

[0763] R s and R s 'Each independently is H, C 1-10 Alkyl, -L d -OR d or -L d -NR d R' d ;

[0764] R' is H, C 1-14 Alkyl, -L a -OR a or -L a -NR a R' a ;

[0765] R" is H or C 1-14 alkyl;

[0766] L d Does not exist or C 1-10 alkylene;

[0767] L a Does not exist or C 1-14 alkylene;

[0768] R a and R' a Each independently is H, C 1-14 Alkyl, C 3-10 cycloalkyl or 3 to 10 membered heterocyclyl, and wherein the alkyl, cycloalkyl and heterocyclyl are optionally substituted;

[0769] R d and R' d are each independently H or C 1-10 alkyl;

[0770] a' and b are each independently 0, 1, 3, 4, or 5, provided that at least one of a' and b is not 0;

[0771] g is 0, 1, 2, 3, 4, or 5;

[0772] a'+g is 0, 1, 2, 3, 4 or 5; and

[0773] c, d, e and f are each independently 0, 1, 2, 3, 4, 5, 6 or 7, with the proviso that c+d is an integer between 2 and 9, and e+f is an integer between 2 and 9.

[0774] In one embodiment, R1 is C 4-20 In one embodiment, R1 is C 6-18In one embodiment, R1 is C 8-18 In one embodiment, R1 is C8, C9, C 10 、C 11 、C 12 、C 13 、C 14 、C 15 、C 16 、C 17 、C 18 、C 19 or C 20 In one embodiment, R1 is C 4-20 In one embodiment, R1 is C 6-18 In one embodiment, R1 is C 8-18 In one embodiment, R1 is C8, C9, C 10 、C 11 、C 12 、C 13 、C 14 、C 15 、C 16 、C 17 、C 18 、C 19 or C 20 In one embodiment, R1 is C 4-20 In one embodiment, R1 is C 6-18 In one embodiment, R1 is C 8-18 In one embodiment, R1 is C8, C9, C 10 、C 11 、C 12 、C 13 、C 14 、C 15 、C 16 、C 17 、C 18 、C 19 or C 20 Alkynyl.

[0775] In one embodiment, one or more -CH2- groups in R1 are replaced by -NH-. In one embodiment, one or more -CH2- groups in R1 are replaced by -N(C 1-20 In one embodiment, one or more -CH2- groups in R1 are replaced by -N(C 1-12 alkyl)-replacement.

[0776] In one embodiment, R1 is unsubstituted. In one embodiment, R1 is replaced by -L a -ORa In one embodiment, R1 is replaced by -L a -SR a In one embodiment, R1 is replaced by -L a -NR a R' a replace.

[0777] In one embodiment, R2 is C 4-20 In one embodiment, R2 is C 6-18 In one embodiment, R2 is C 8-18 In one embodiment, R2 is C8, C9, C 10 、C 11 、C 12 、C 13 、C 14 、C 15 、C 16 、C 17 、C 18 、C 19 or C 20 In one embodiment, R2 is C 4-20 In one embodiment, R2 is C 6-18 In one embodiment, R2 is C 8-18 In one embodiment, R2 is C8, C9, C 10 、C 11 、C 12 、C 13 、C 14 、C 15 、C 16 、C 17 、C 18 、C 19 or C 20 In one embodiment, R2 is C 4-20 In one embodiment, R2 is C 6-18 In one embodiment, R2 is C 8-18 In one embodiment, R2 is C8, C9, C 10 、C 11 、C 12 、C 13 、C 14 、C 15 、C 16 、C 17 、C 18 、C 19 or C 20 Alkynyl.

[0778] In one embodiment, one or more -CH2- groups in R2 are replaced by -NH-. In one embodiment, one or more -CH2- groups in R2 are replaced by -N(C 1-20 In one embodiment, one or more -CH2- groups in R2 are replaced by -N(C 1-12 alkyl)-replacement.

[0779] In one embodiment, R2 is unsubstituted. In one embodiment, R2 is -L a -OR a In one embodiment, R2 is replaced by -L a -SR a In one embodiment, R2 is replaced by -L a -NR a R' a replace.

[0780] In one embodiment, L a In one embodiment, L a It is C 1-14 In one embodiment, L a It is C 1-6 In one embodiment, L a In one embodiment, L a It is ethylene.

[0781] In one embodiment, R a It is C 1-14 In one embodiment, R a is a C3-10 cycloalkyl group. a is a 3- to 10-membered heterocyclyl. a It is C 1-10 Alkyl; in one embodiment, R a It is C 8-10 Alkyl; in one embodiment, R a It is C 8-10 Straight chain alkyl; in one embodiment, R a is -(CH2)8CH3; in one embodiment, R a Optionally substituted with one or more of the following substituents: H, C 1-20 Alkyl, -L e -OR e 、-L e -SR e and -L e -NR e R' e .

[0782] In one embodiment, R' a It is C 1-14 In one embodiment, R' a It is C 3-10 In one embodiment, R' a In one embodiment, R' a It is C 8-10 Alkyl; in one embodiment, R' a It is C 8-10 linear alkyl; in one embodiment, R' a is -(CH2)8CH3; in one embodiment, R' a Optionally substituted with one or more of the following substituents: H, C 1-20 Alkyl, -L e -OR e 、-L e -SR e and -L e -NR e R' e .

[0783] In one embodiment, R a and R' a Together with the nitrogen to which they are attached, they form a 4 to 10 membered ring. In one embodiment, R a and R' a Together with the nitrogen to which they are attached, they form a 4 to 8 membered ring. a and R' a Together with the nitrogen to which they are attached, they form a 4 to 6 membered ring. a and R' a Together with the nitrogen to which they are attached, they form a 4 to 6 membered cycloalkyl group. a and R' a Together with the nitrogen to which they are attached they form a 4- to 6-membered heterocyclic group.

[0784] In one embodiment, R3 is H. In one embodiment, R3 is C 1-10 In one embodiment, R3 is C 1-10 In one embodiment, R3 is C 2-10 In one embodiment, R3 is C 2-10 In one embodiment, R3 is a 3 to 14 membered cycloalkyl. In one embodiment, R3 is a 3 to 14 membered heterocyclyl. In one embodiment, R3 is C 6-10In one embodiment, R3 is a 5- to 14-membered heteroaryl. In one embodiment, R3 is C 1-6 In one embodiment, R3 is C 1-6 In one embodiment, R3 is a 3- to 10-membered cycloalkyl group. In one embodiment, R3 is a 3- to 10-membered heterocyclyl group. In one embodiment, R3 is a 3- to 7-membered cycloalkyl group. In one embodiment, R3 is a 3- to 7-membered heterocyclyl group.

[0785] In one embodiment, R3 is unsubstituted. In one embodiment, R3 is substituted with one or more R*, wherein each R* is independently halogen, cyano, C 1-10 Alkyl, C 1-10 Halogenated alkyl, -L b -ORb, -L b -SRb or -L b -NRbR'b. In one embodiment, R3 is optionally substituted with 1, 2, 3, 4 or 5 R*, wherein each R* is independently halogen, cyano, C 1-10 Alkyl, C 1-10 Halogenated alkyl, -L b -ORb, -L b -SRb or -L b -NRbR'b. In one embodiment, R3 is Me. In one embodiment, R3 is -CH2CH3. In one embodiment, R3 is -CH(CH3)2.

[0786] In one embodiment, R3 is C optionally substituted with OH 1-6 In one embodiment, R3 is -CH2CH2OH.

[0787] In one embodiment, R4 is H. In one embodiment, R4 is C 1-10 In one embodiment, R4 is C 1-10 In one embodiment, R4 is C 2-10 In one embodiment, R4 is C 2-10 In one embodiment, R4 is a 3 to 14 membered cycloalkyl. In one embodiment, R4 is a 3 to 14 membered heterocyclyl. In one embodiment, R4 is C 6-10 In one embodiment, R4 is a 5- to 14-membered heteroaryl. In one embodiment, R4 is C 1-6 In one embodiment, R4 is C 1-6haloalkyl. In one embodiment, R4 is a 3- to 10-membered cycloalkyl. In one embodiment, R4 is a 3- to 10-membered heterocyclyl. In one embodiment, R4 is a 3- to 7-membered cycloalkyl. In one embodiment, R4 is a 3- to 7-membered heterocyclyl. In one embodiment, R4 is Me. In one embodiment, R3 is unsubstituted. In one embodiment, R3 is substituted with one or more R*. In one embodiment, R3 is optionally substituted with 1, 2, 3, 4, or 5 R*.

[0788] In one embodiment, R3 and R4 together with the N atom to which they are attached form a 3- to 14-membered heterocyclyl. In one embodiment, R3, R4 together with the N atom to which they are attached form a 3- to 10-membered heterocyclyl; in one embodiment, R3, R4 together with the N atom to which they are attached form a 3- to 7-membered heterocyclyl; in one embodiment, R3, R4 together with the N atom to which they are attached form a 5- to 7-membered heterocyclyl; in one embodiment, R3, R4 together with the N atom to which they are attached form a 4- to 6-membered heterocyclyl; in one embodiment, R3, R4 together with the N atom to which they are attached form a 5-membered heterocyclyl; in one embodiment, R3, R4 together with the N atom to which they are attached form a In one embodiment, R3, R4 together with the N atom to which they are attached form In one embodiment, R3, R4 together with the N atom to which they are attached form In one embodiment, R3, R4 together with the N atom to which they are attached form In one embodiment, the heterocyclyl formed by R3 and R4 together with the N atom to which they are attached is optionally substituted with one or more R*; in one embodiment, the heterocyclyl formed by R3 and R4 together with the N atom to which they are attached is optionally substituted with 1, 2, 3, 4 or 5 R*.

[0789] In one embodiment, R4 together with the nitrogen atom to which it is attached and an R0' forms a 3 to 14-membered heterocyclic radical. In one embodiment, R4 together with the nitrogen atom to which it is attached and an R0' forms a 4 to 10-membered heterocyclic radical. In one embodiment, R4 together with the nitrogen atom to which it is attached and an R0' forms a 4 to 6-membered heterocyclic radical. In one embodiment, R4 together with the nitrogen atom to which it is attached and an R0' forms a 5 to 14-membered heteroaryl. In one embodiment, R4 together with the nitrogen atom to which it is attached and an R0' forms a 5 to 10-membered heteroaryl. In one embodiment, R4 together with the nitrogen atom to which it is attached and an R0' forms a 5 or 6-membered heteroaryl.

[0790] In one embodiment, R5 is H; in one embodiment, R5 is C1-8 Alkyl; in one embodiment, R5 is C 1-6 Alkyl; in one embodiment, R5 is C 1-3 Alkyl; in one embodiment, R5 is Me; in one embodiment, R5 is optionally substituted with one or more R*; in one embodiment, R5 is optionally substituted with 1, 2, 3, 4 or 5 R*.

[0791] In one embodiment, R6 is H; in one embodiment, R6 is C 1-8 Alkyl; in one embodiment, R6 is C 1-6 Alkyl; in one embodiment, R6 is C 1-3 Alkyl; in one embodiment, R6 is Me; in one embodiment, R6 is optionally substituted with one or more R*; in one embodiment, R6 is optionally substituted with 1, 2, 3, 4 or 5 R*.

[0792] In one embodiment, R5 and R6 together with the carbon atom to which they are attached form C 3-14 Cycloalkylene or 3- to 14-membered heterocyclic ring, wherein the cycloalkylene and heterocyclic ring are optionally replaced by one or more R 4s In one embodiment, R5 and R6 together with the carbon atom to which they are attached form C 3-8 In one embodiment, R5 and R6 together with the carbon atom to which they are attached form C 3-6 Cycloalkylene. In one embodiment, R5 and R6 together with the carbon atom to which they are attached form a C3 cycloalkylene, C4 cycloalkylene, C5 cycloalkylene or C6 cycloalkylene. In one embodiment, R5 and R6 together with the carbon atom to which they are attached form a 3- to 8-membered heterocyclic ring. In one embodiment, R5 and R6 together with the carbon atom to which they are attached form a 3- to 6-membered heterocyclic ring. In one embodiment, R5 and R6 together with the carbon atom to which they are attached form a 3-membered heterocyclic ring, a 4-membered heterocyclic ring, a 5-membered heterocyclic ring or a 6-membered heterocyclic ring. In one embodiment, the heterocyclic ring has one or more oxygen, sulfur or nitrogen atoms on the ring. In one embodiment, the heterocyclic ring or cycloalkylene formed by R5 and R6 is unsubstituted. In one embodiment, the heterocyclic ring or cycloalkylene formed by R5 and R6 is substituted by 1, 2, 3, 4 or 5 R*. In one embodiment, the heterocyclic ring or cycloalkylene formed by R5 and R6 is substituted by 1, 2, 3, 4 or 5 R 4s replace.

[0793] In one embodiment, R7 is H; in one embodiment, R7 is C 1-8 Alkyl; in one embodiment, R7 is C 1-6Alkyl; in one embodiment, R7 is C 1-3 Alkyl; in one embodiment, R7 is Me; in one embodiment, R7 is optionally substituted with one or more R*; in one embodiment, R7 is optionally substituted with 1, 2, 3, 4 or 5 R*.

[0794] In one embodiment, R5, R6, R7 and R8 are each independently hydrogen or methyl. In one embodiment, R5 and R6 are each independently C 1-6 In one embodiment, R5 and R6 are methyl, and R7 and R8 are hydrogen. In one embodiment, R5, R6, R7, R8 are hydrogen.

[0795] In one embodiment, R8 is H; in one embodiment, R8 is C 1-8 Alkyl; in one embodiment, R8 is C 1-6 Alkyl; in one embodiment, R8 is C 1-3 Alkyl; in one embodiment, R8 is Me; in one embodiment, R8 is optionally substituted with one or more R*; in one embodiment, R8 is optionally substituted with 1, 2, 3, 4 or 5 R*.

[0796] In one embodiment, R7 and R8 together with the carbon atom to which they are attached form C 3-14 Cycloalkylene or 3- to 14-membered heterocyclic ring, wherein the cycloalkylene and heterocyclic ring are optionally replaced by one or more R 4s In one embodiment, R7 and R8 together with the carbon atom to which they are attached form a C3-8 cycloalkylene group. In one embodiment, R7 and R8 together with the carbon atom to which they are attached form a C3-8 cycloalkylene group. 3-6 Cycloalkylene. In one embodiment, R7 and R8, together with the carbon atom to which they are attached, form a C3 cycloalkylene, C4 cycloalkylene, C5 cycloalkylene or C6 cycloalkylene. In one embodiment, R7 and R8, together with the carbon atom to which they are attached, form a 3- to 8-membered heterocyclic ring. In one embodiment, R7 and R8, together with the carbon atom to which they are attached, form a 3- to 6-membered heterocyclic ring. In one embodiment, R7 and R8, together with the carbon atom to which they are attached, form a 3-membered heterocyclic ring, a 4-membered heterocyclic ring, a 5-membered heterocyclic ring or a 6-membered heterocyclic ring. In one embodiment, the heterocyclic ring has one or more oxygen, sulfur or nitrogen atoms on the ring. In one embodiment, the heterocyclic ring or cycloalkylene formed by R7 and R8 is unsubstituted. In one embodiment, the heterocyclic ring or cycloalkylene formed by R7 and R8 is substituted by 1, 2, 3, 4 or 5 R*. In one embodiment, the heterocyclic ring or cycloalkylene formed by R7 and R8 is substituted by 1, 2, 3, 4 or 5 R*. 4s replace.

[0797] In one embodiment, R* is halogen, cyano, C 1-6 Alkyl, C 1-6 Halogenated alkyl, -L b -OR b or -L b -NR b R' b In one embodiment, R* is C 1-6 Alkyl, C 1-6 Haloalkyl or -OR b In one embodiment, R* is independently H, halogen, C 1-6 Alkyl or C 1-6 In one embodiment, R* is C 1-6 Alkyl or C 1-6 In one embodiment, R* is halogen; In one embodiment, R* is cyano; In one embodiment, R* is C 1-10 Alkyl; in one embodiment, R* is C 1-10 haloalkyl; in one embodiment, R* is -L b -OR b ; In one embodiment, R* is -L b -SR b ; In one embodiment, R* is -L b -NR b R' b In one embodiment, R* is C 1-6 Alkyl; in one embodiment, R* is C 1-6 haloalkyl; in one embodiment, R* is -OR b .

[0798] In one embodiment, L b In one embodiment, L b It is C 1-10 In one embodiment, L b It is C 1-6 In one embodiment, L b It is C 1-3 In one embodiment, L b In one embodiment, L b It is ethylene.

[0799] In one embodiment, L f In one embodiment, L f It is C1-10 In one embodiment, L f It is C 1-6 In one embodiment, L f It is C 1-3 In one embodiment, L f In one embodiment, L f It is ethylene.

[0800] In one embodiment, L c In one embodiment, L c It is C 1-8 In one embodiment, L c It is C 1-6 In one embodiment, L c It is C 1-3 In one embodiment, L c In one embodiment, L c It is ethylene.

[0801] In one embodiment, R b It is C 1-10 In one embodiment, R b It is C 1-6 In one embodiment, R b It is C 1-3 In one embodiment, R b It is C 3-14 In one embodiment, R b It is C 3-8 In one embodiment, R b It is C 3-6 In one embodiment, R b is a 3 to 14 membered heterocyclyl. In one embodiment, R b is a 3 to 8 membered heterocyclyl. b is a 3- to 6-membered heterocyclic group. b is a 3 to 6-membered nitrogen-containing heterocyclic group. b In one embodiment, R b C 1-10 Alkyl, -L f -OR f 、-L f -SR f or -L f -NR f R' f One or more substitutions, wherein Rf and R' f are each independently H or C 1-10 alkyl.

[0802] In one embodiment, R' b It is C 1-10 In one embodiment, R' b It is C 1-6 In one embodiment, R' b It is C 1-3 In one embodiment, R' b It is C 3-14 In one embodiment, R' b It is C 3-8 In one embodiment, R' b It is C 3-6 In one embodiment, R' b is a 3 to 14 membered heterocyclyl. In one embodiment, R' b is a 3 to 8 membered heterocyclic group. In one embodiment, R' b is a 3 to 6 membered heterocyclic group. In one embodiment, R' b is a 3 to 6-membered nitrogen-containing heterocyclic group. In one embodiment, R' b is unsubstituted. In one embodiment, R' b C 1-10 Alkyl, -L f -OR f 、-L f -SR f or -L f -NR f R' f One or more substitutions, wherein R f and R' f are each independently H or C 1-10 alkyl.

[0803] In one embodiment, j is 0. In one embodiment, j is 1.

[0804] In one embodiment, W is CH. In one embodiment, W is N.

[0805] In one embodiment, k is 0. In one embodiment, k is 1. In one embodiment, k is 2.

[0806] In one embodiment, k is 3. In one embodiment, k is 4. In one embodiment, k is 5. In one embodiment, k is 6. In one embodiment, k is 7. In one embodiment, k is 8.

[0807] In one embodiment, the two substituents of R0' together with the methylene group to which they are attached form a 3 to 8 membered cycloalkyl. In one embodiment, the two substituents of R0' together with the methylene group to which they are attached form a 3 to 6 membered cycloalkyl. In one embodiment, the two substituents of R0' together with the methylene group to which they are attached form a 5 or 6 membered cycloalkyl.

[0808] In one embodiment, M1 is -C(O)O-; in one embodiment, M1 is -O-; in one embodiment, M1 is -SC(O)O-; in one embodiment, M1 is -OC(O)NR a -; In one embodiment, M1 is -NR a C(O)NR a -; In one embodiment, M1 is -OC(O)S-; In one embodiment, M1 is -OC(O)O-; In one embodiment, M1 is -NR a C(O)O-; in one embodiment, M1 is -OC(O)-; in one embodiment, M1 is -SC(O)-; in one embodiment, M1 is -C(O)S-; in one embodiment, M1 is -NR a -; In one embodiment, M1 is -C(O)NR a -; In one embodiment, M1 is -NR a C(O)-; in one embodiment, M1 is -NR a C(O)S-; in one embodiment, M1 is -SC(O)NR a -; in one embodiment, M1 is -C(O)-; in one embodiment, M1 is -OC(S)-; in one embodiment, M1 is -C(S)O-; in one embodiment, M1 is -OC(S)NR a -; In one embodiment, M1 is -NR a C(S)O-; in one embodiment, M1 is -SS-; in one embodiment, M1 is -S(O) 0-2 -.

[0809] In one embodiment, M2 is -C(O)O-; in one embodiment, M2 is -O-; in one embodiment, M2 is -SC(O)O-; in one embodiment, M2 is -OC(O)NR a -; In one embodiment, M2 is -NR a C(O)NR a-; In one embodiment, M2 is -OC(O)S-; In one embodiment, M2 is -OC(O)O-; In one embodiment, M2 is -NR a C(O)O-; in one embodiment, M2 is -OC(O)-; in one embodiment, M2 is -SC(O)-; in one embodiment, M2 is -C(O)S-; in one embodiment, M2 is -NR a -; In one embodiment, M2 is -C(O)NR a -; In one embodiment, M2 is -NR a C(O)-; in one embodiment, M2 is -NR a C(O)S-; in one embodiment, M2 is -SC(O)NR a -; In one embodiment, M2 is -C(O)-; In one embodiment, M2 is -OC(S)-; In one embodiment, M2 is -C(S)O-; In one embodiment, M2 is -OC(S)NR a -; In one embodiment, M2 is -NR a C(S)O-; in one embodiment, M2 is -SS-; in one embodiment, M2 is -S(O) 0-2 -.

[0810] In one embodiment, M1 and M2 are each independently selected from -C(O)O-, -SC(O)O-, -OC(O)NR a -、 -NR a C(O)NR a -、-OC(O)S-、-OC(O)O-、-NR a C(O)O-, -C(O)S-, -C(O)NR a -、-NR a C(O)S-、-SC(O)NR a -、-C(S)O-、-OC(S)NR a -and-NR a C(S)O-; In one embodiment, M1 and M2 are independently -C(O)O-, -C(O)S-, -C(O)NR a - or -C(S)O-; in one embodiment, M1 and M2 are independently -C(O)O-, -C(O)S- or -C(O)NR a In one embodiment, M1 and M2 are both -C(O)O-.

[0811] In one embodiment, Q is absent; in another embodiment, Q is -C(O)O-; in another embodiment, Q is -O-; in another embodiment, Q is -SC(O)O-; in another embodiment, Q is -OC(O)NR b -; In another embodiment, Q is -NR b C(O)NR b -; in another embodiment, Q is -OC(O)S-; in another embodiment, Q is -OC(O)O-; in another embodiment, Q is -NR b C(O)O-; in another embodiment, Q is -OC(O)-; in another embodiment, Q is -SC(O)-; in another embodiment, Q is -C(O)S-; in another embodiment, Q is -NR b -; In another embodiment, Q is -C(O)NR b -; In another embodiment, Q is -NR b C(O)-; In another embodiment, Q is -NR b C(O)S-; in another embodiment, Q is -SC(O)NR b -; in another embodiment, Q is -C(O)-; in another embodiment, Q is -OC(S)-; in another embodiment, Q is -C(S)O-; in another embodiment, Q is -OC(S)NR b -; In another embodiment, Q is -NR b C(S)O-; in another embodiment, Q is -SS-; in another embodiment, Q is -S(O) 0-2 -; in another embodiment, Q is phenylene; in another embodiment, Q is pyridinylene; in another embodiment, phenylene or pyridinylene is optionally substituted with one or more R*.

[0812] In one embodiment, Q is selected from absent, -C(O)O-, -O-, -SC(O)O-, -OC(O)NR b -、-NR b C(O)NR b -、-OC(O)S-、-OC(O)O-、-NR b C(O)O-, -OC(O)-, -SC(O)-, -C(O)S-, -NR b -、-C(O)NR b -、-NR b C(O)-、-NR b C(O)S-、-SC(O)NR b-, -C(O)-, -OC(S)-, -C(S)O-, -OC(S)NR b -、-NR b C(S)O-, -SS-, and -S(O) 0-2 -; In one embodiment, Q is selected from -C(O)O-, -O-, -SC(O)O-, -OC(O)NH-, -NHC(O)NH-, -OC(O)S-, -OC(O)O-, -NHC(O)O-, -OC(O)-, -SC(O)-, -C(O)S-, -NH-, -C(O)NH-, -NHC(O)-, -NHC(O)S-, -SC(O) In one embodiment, Q is selected from the group consisting of -C(O)O-, -O-, -SC(O)O-, -OC(O)NH-, -NHC(O)NH-, -OC(O)S-, -OC(O)O-, and -NHC(O)O-; in one embodiment, Q is -C(O)O-.

[0813] In one embodiment, R a is H; in another embodiment, R a It is C 1-20 Alkyl; in another embodiment, R a is a 3- to 14-membered cycloalkyl group; in another embodiment, R a is a 3- to 14-membered heterocyclyl; in another embodiment, R a It is C 1-14 Alkyl; in another embodiment, R a It is C 1-10 Alkyl; in another embodiment, R a It is C 8-10 Alkyl; in another embodiment, R a It is C 8-10 linear alkyl; in another embodiment, R a is -(CH2)8CH3; in another embodiment, R a Optionally substituted with one or more of the following substituents: H, C 1-20 Alkyl, -L e -OR e 、 -L e -SR e and -L e -NR e R' e , where L e Does not exist or C 1-20 Alkylene, where R e and R'e are independently H or C 1-20 alkyl;

[0814] In one embodiment, R' a is H; in another embodiment, R' a It is C 1-20 Alkyl; in another embodiment, R' a is a 3 to 14 membered cycloalkyl group; in another embodiment, R' a is a 3- to 14-membered heterocyclyl; in another embodiment, R' a It is C 1-14 Alkyl; in another embodiment, R' a It is C 1-10 Alkyl; in another embodiment, R' a It is C 8-10 Alkyl; in another embodiment, R' a It is C 8-10 linear alkyl; in another embodiment, R' a is -(CH2)8CH3; in another embodiment, R' a Optionally substituted with one or more of the following substituents: H, C 1-20 Alkyl, -L e -OR e 、-L e -SR e and -L e -NR e R' e , where L e Does not exist or C 1-20 Alkylene, where R e and R' e are independently H or C 1-20 alkyl.

[0815] In one embodiment, L e In one embodiment, L e It is C 1-20 In one embodiment, L e It is C 1-16 In one embodiment, L e It is C 1-12 In one embodiment, L e It is C 1-6 In one embodiment, L e In one embodiment, L e It is ethylene.

[0816] In one embodiment, Re In one embodiment, R e It is C 1-20 In one embodiment, R e It is C 1-16 In one embodiment, R e It is C 1-12 In one embodiment, R e It is C 1-6 In one embodiment, R' e In one embodiment, R' e It is C 1-20 In one embodiment, R' e It is C 1-16 In one embodiment, R' e It is C 1-12 In one embodiment, R' e It is C 1-6 alkyl.

[0817] In one embodiment, R f In one embodiment, R f It is C 1-10 In one embodiment, R f It is C 1-6 In one embodiment, R f It is C 1-3 In one embodiment, R' f In one embodiment, R' f It is C 1-10 In one embodiment, R' f It is C 1-6 In one embodiment, R' f It is C 1-3 alkyl.

[0818] In one embodiment, G1 is absent; in another embodiment, G1 is C 1-13 Alkylene; in another embodiment, G1 is C 2-13 Alkenylene; in another embodiment, G1 is C 2-6 Alkenylene; in another embodiment, G1 is C 2-13 Alkyne; in another embodiment, G1 is C 2-6 Alkyne; In another embodiment, G1 is optionally replaced by one or more R s Substituted, where R s Independently H, C 1-14 Alkyl, -L d -ORd 、-L d -SR d or -L d -NR d R' d , and where L d Does not exist or C 1-14 alkylene; and wherein R d and R' d are independently H or C 1-14 alkyl.

[0819] In one embodiment, G2 is absent; in another embodiment, G2 is C 2-13 Alkylene; in another embodiment, G2 is C 2-6 Alkylene; in another embodiment, G2 is C 2-13 Alkenylene; in another embodiment, G2 is C 2-6 Alkenylene; in another embodiment, G2 is C 2-13 Alkyne; In another embodiment, G2 is optionally replaced by one or more R s Substituted, where R s Independently H, C 1-14 Alkyl, -L d -OR d 、-L d -SR d or -L d -NR d R' d , and where L d Does not exist or C 1-14 alkylene; and wherein R d and R' d are independently H or C 1-14 alkyl.

[0820] In one embodiment, the total length of G1 and G2 is 3 carbon atoms; in another embodiment, the total length of G1 and G2 is 4 carbon atoms; in another embodiment, the total length of G1 and G2 is 5 carbon atoms; in another embodiment, the total length of G1 and G2 is 6 carbon atoms; in another embodiment, the total length of G1 and G2 is 7 carbon atoms; in another embodiment, the total length of G1 and G2 is 8 carbon atoms; in another embodiment, the total length of G1 and G2 is 9 carbon atoms; in another embodiment, the total length of G1 and G2 is 10 carbon atoms; in another embodiment, the total length of G1 and G2 is 11 carbon atoms; in another embodiment, the total length of G1 and G2 is 12 carbon atoms; in another embodiment, the total length of G1 and G2 is 13 carbon atoms.

[0821] In one embodiment, G3 is absent; in another embodiment, G3 is C 1-13 Alkylene; in another embodiment, G3 is C 2-13 Alkenylene; in another embodiment, G3 is C 2-6 Alkenylene; in another embodiment, G3 is C 2-13 Alkyne; in another embodiment, G3 is C 2-6 Alkyne; In another embodiment, G3 is optionally replaced by one or more R s Substituted, where R s Independently H, C 1-14 Alkyl, -L d -OR d 、-L d -SR d or -L d -NR d R' d , and where L d Does not exist or C 1-14 alkylene; and wherein R d and R' d are independently H or C 1-14 alkyl.

[0822] In one embodiment, G4 is absent; in another embodiment, G4 is C 2-13 Alkylene; in another embodiment, G4 is C 2-6 Alkylene; in another embodiment, G4 is C 2-13 Alkenylene; in another embodiment, G4 is C 2-6 Alkenylene; in another embodiment, G4 is C 2-13 Alkyne; In another embodiment, G4 is optionally replaced by one or more R s Substituted, where R s Independently H, C 1-14 Alkyl, -L d -OR d 、-L d -SR d or -L d -NR d R' d , and where L d Does not exist or C 1-14 alkylene; and wherein R d and R' d are independently H or C 1-14 alkyl.

[0823] In one embodiment, G1, G2, G3 and G4 are each independently absent or C 1-8 Alkylene.

[0824] In one embodiment, the total length of G3 and G4 is 3 carbon atoms; in another embodiment, the total length of G3 and G4 is 4 carbon atoms; in another embodiment, the total length of G3 and G4 is 5 carbon atoms; in another embodiment, the total length of G3 and G4 is 6 carbon atoms; in another embodiment, the total length of G3 and G4 is 7 carbon atoms; in another embodiment, the total length of G3 and G4 is 8 carbon atoms; in another embodiment, the total length of G3 and G4 is 9 carbon atoms; in another embodiment, the total length of G3 and G4 is 10 carbon atoms; in another embodiment, the total length of G3 and G4 is 11 carbon atoms; in another embodiment, the total length of G3 and G4 is 12 carbon atoms; in another embodiment, the total length of G3 and G4 is 13 carbon atoms.

[0825] In one embodiment, R s is H; in another embodiment, R s It is C 1-14 Alkyl; in another embodiment, R s Yes-L d -OR d In another embodiment, R s Yes-L d -SR d In another embodiment, R s Yes-L d -NR d R' d In another embodiment, R s It is C 1-10 Alkyl; in another embodiment, R s It is C 1-6 In one embodiment, R s It is H, C 1-10 Alkyl, -L d -OR d or -L d -NR d R' d In another more specific embodiment, R s Is H or C 1-6 alkyl.

[0826] In one embodiment, G5 is absent. In one embodiment, G5 is absent, j is 0, and R3 is CH2CH2OH.

[0827] In another embodiment, G5 is C 1-8 Alkylene; in another embodiment, G5 is C 1-6 Alkylene; in another embodiment, G5 is C 1-3 In another embodiment, G5 is optionally substituted with one or more R**, wherein each R** is independently C 1-8 Alkyl, -L c -OR c 、-Lc-SR c or -L c -NRcR' c , where R c and R' c are independently H or C 1-8 alkyl, and wherein L c Does not exist or C 1-6 alkylene;

[0828] In one embodiment, R** is C 1-8 Alkyl; in another embodiment, R** is -L c -OR c In another embodiment, R** is -L c -SR c In another embodiment, R** is -L c -NR c R' c In another embodiment, R** is C 1-6 alkyl.

[0829] In one embodiment, L1 is -(CRR')2-. In one embodiment, L1 is -CH=CH-. In one embodiment, L1 is -C≡C-. In one embodiment, L1 is -NR"-;

[0830] In one embodiment, L2 is -(CRR')2-. In one embodiment, L2 is -CH=CH-. In one embodiment, L2 is -C≡C-. In one embodiment, L2 is -NR"-;

[0831] In one embodiment, L3 is -(CR s R s ')2-, and L5 is absent. In one embodiment, L3 is -CH=CH-, and L5 is absent. In one embodiment, L3 is -C≡C-, and L5 is absent. In one embodiment, L5 is -(CR s R s')2-, and L3 is absent. In one embodiment, L5 is -CH=CH-, and L3 is absent. In one embodiment, L5 is -C≡C-, and L3 is absent.

[0832] In one embodiment, L4 is -(CR s R s ')2-, and L6 is absent. In one embodiment, L4 is -CH=CH-, and L6 is absent. In one embodiment, L4 is -C≡C-, and L6 is absent. In one embodiment, L6 is -(CR s R s ')2-, and L4 is absent. In one embodiment, L6 is -CH=CH-, and L4 is absent. In one embodiment, L6 is -C≡C-, and L4 is absent.

[0833] In one embodiment, G 1a In one embodiment, G 1a In one embodiment, G 1a In one embodiment, G 1a In one embodiment, G 1a In one embodiment, G 1a In one embodiment, G 1a In one embodiment, G 1a In one embodiment, G 1a In one embodiment, G 1a In one embodiment, G 1a By one or more R s Substitute, where each R s Independently H, C 1-14 Alkyl, -L d -OR d 、-L d -SR d or -L d -NR d R' d , and where L d Does not exist or C 1-14 alkylene; and wherein R d and R' d are independently H or C 1-14 alkyl.

[0834] In one embodiment, G 1bIn one embodiment, G 1b In one embodiment, G 1b In one embodiment, G 1b In one embodiment, G 1b In one embodiment, G 1b In one embodiment, G 1b In one embodiment, G 1b In one embodiment, G 1b In one embodiment, G 1b In one embodiment, G 1b By one or more R s Substitute, where each R s Independently H, C 1-14 Alkyl, -L d -OR d 、-L d -SR d or -L d -NR d R' d , and where L d Does not exist or C 1-14 alkylene; and wherein R d and R' d are independently H or C 1-14 alkyl.

[0835] In one embodiment, G 2a In one embodiment, G 2a In one embodiment, G 2a In one embodiment, G 2a In one embodiment, G 2a In one embodiment, G 2a In one embodiment, G 2a In one embodiment, G 2a In one embodiment, G 2a In one embodiment, G 2a In one embodiment, G 2a By one or more R s Substitute, where each R s Independently H, C 1-14 Alkyl, -L d -ORd 、-L d -SR d or -L d -NR d R' d , and where L d Does not exist or C 1-14 alkylene; and wherein R d and R' d are independently H or C 1-14 alkyl.

[0836] In one embodiment, G 2b In one embodiment, G 2b In one embodiment, G 2b In one embodiment, G 2b In one embodiment, G 2b In one embodiment, G 2b In one embodiment, G 2b In one embodiment, G 2b In one embodiment, G 2b In one embodiment, G 2b In one embodiment, G 2b By one or more R s Substitute, where each R s Independently H, C 1-14 Alkyl, -L d -OR d 、-L d -SR d or -L d -NR d R' d , and where L d Does not exist or C 1-14 alkylene; and wherein R d and R' d are independently H or C 1-14 alkyl.

[0837] In one embodiment, G 3a In one embodiment, G 3a In one embodiment, G 3a In one embodiment, G 3a In one embodiment, G 3a In one embodiment, G 3aIn one embodiment, G 3a In one embodiment, G 3a In one embodiment, G 3a In one embodiment, G 3a In one embodiment, G 3a By one or more R s Substitute, where each R s Independently H, C 1-14 Alkyl, -L d -OR d 、-L d -SR d or -L d -NR d R' d , and where L d Does not exist or C 1-14 alkylene; and wherein R d and R' d are independently H or C 1-14 alkyl.

[0838] In one embodiment, G 3b In one embodiment, G 3b In one embodiment, G 3b In one embodiment, G 3b In one embodiment, G 3b In one embodiment, G 3b In one embodiment, G 3b In one embodiment, G 3b In one embodiment, G 3b In one embodiment, G 3b In one embodiment, G 3b By one or more R s Substitute, where each R s Independently H, C 1-14 Alkyl, -L d -OR d 、-L d -SR d or -L d -NR d R' d , and where L d Does not exist or C 1-14 alkylene; and wherein R dand R' d are independently H or C 1-14 alkyl.

[0839] In one embodiment, G 4a In one embodiment, G 4a In one embodiment, G 4a In one embodiment, G 4a In one embodiment, G 4a In one embodiment, G 4a In one embodiment, G 4a In one embodiment, G 4a In one embodiment, G 4a In one embodiment, G 4a In one embodiment, G 4a By one or more R s Substitute, where each R s Independently H, C 1-14 Alkyl, -L d -OR d 、-L d -SR d or -L d -NR d R' d , and where L d Does not exist or C 1-14 alkylene; and wherein R d and R' d are independently H or C 1-14 alkyl.

[0840] In one embodiment, G 4b In one embodiment, G 4b In one embodiment, G 4b In one embodiment, G 4b In one embodiment, G 4b In one embodiment, G 4b In one embodiment, G 4b In one embodiment, G 4b In one embodiment, G 4b In one embodiment, G 4b In one embodiment, G 4bBy one or more R s Substitute, where each R s Independently H, C 1-14 Alkyl, -L d -OR d 、-L d -SR d or -L d -NR d R' d , and where L d Does not exist or C 1-14 alkylene; and wherein R d and R' d are independently H or C 1-14 alkyl.

[0841] In one embodiment, G 1a , G 1b , G 2a and G 2b The total length of G is 1, 2, 3, 4, 5, 6 or 7 carbon atoms; in another more specific embodiment, G 1a , G 1b , G 2a and G 2b The total length of G is 1, 2, 3, 4, 5 or 6 carbon atoms. 1a , G 1b , G 2a and G 2b The total length of G is 1 carbon atom. 1a , G 1b , G 2a and G 2b The total length of G is 2 carbon atoms. 1a , G 1b , G 2a and G 2b The total length of G is 3 carbon atoms. 1a , G 1b , G 2a and G 2b The total length of G is 4 carbon atoms. 1a , G 1b , G 2a and G 2b The total length of G is 5 carbon atoms. 1a , G 1b , G 2a and G 2b The total length is 6 carbon atoms.

[0842] In one embodiment, G3a , G 3b , G 4a and G 4b The total length of G is 1, 2, 3, 4, 5, 6 or 7 carbon atoms. 3a , G 3b , G 4a and G 4b The total length of G is 1 carbon atom. 3a , G 3b , G 4a and G 4b The total length of G is 2 carbon atoms. 3a , G 3b , G 4a and G 4b The total length of G is 3 carbon atoms. 3a , G 3b , G 4a and G 4b The total length of G is 4 carbon atoms. 3a , G 3b , G 4a and G 4b The total length of G is 5 carbon atoms. 3a , G 3b , G 4a and G 4b The total length of G is 6 carbon atoms. 3a , G 3b , G 4a and G 4b The total length is 7 carbon atoms.

[0843] In one embodiment, G7 is absent; in another embodiment, G7 is C 1-12 Alkylene; in another embodiment, G7 is C 1-6 In another embodiment, G7 is C1-5 alkylene; In another embodiment, G7 is C1-5 linear alkylene; In another embodiment, G7 is -CH2-; In another embodiment, G7 is -(CH2)2-; In another embodiment, G7 is -(CH2)4-; In another embodiment, G7 is -(CH2)5-; In another embodiment, G7 is optionally substituted with 1, 2, 3, 4, 5 or 6 R, wherein R is C 1-10In another embodiment, 1, 2, or 3 methylene groups in G7 are optionally and independently substituted with 1 R; in another embodiment, 1 or 2 methylene groups in G7 are optionally and independently substituted with 1 R; in another embodiment, the methylene group of G7 connected to M1 is not substituted with R.

[0844] In one embodiment, G8 is absent; in another embodiment, G8 is C 1-12 Alkylene; in another embodiment, G8 is C 1-10 Alkylene; in another embodiment, G8 is C 1-8 Alkylene; in another embodiment, G8 is C 1-8 linear alkylene; in another embodiment, G8 is -(CH2)2-; in another embodiment, G8 is -(CH2)4-; in another embodiment, G8 is -(CH2)6-; in another embodiment, G8 is -(CH2)7-; in another embodiment, G8 is -(CH2)8-; in another embodiment, G8 is optionally substituted with 1, 2, 3, 4, 5, or 6 R, wherein R is C 1-10 alkyl; in another embodiment, 1, 2 or 3 methylene groups in G8 are optionally and independently substituted with 1 R; in another embodiment, 1 or 2 methylene groups in G8 are optionally and independently substituted with 1 R.

[0845] In one embodiment, the total length of G7 and G8 is 4 carbon atoms; in another embodiment, the total length of G7 and G8 is 5 carbon atoms; in another embodiment, the total length of G7 and G8 is 6 carbon atoms; in another embodiment, the total length of G7 and G8 is 7 carbon atoms; in another embodiment, the total length of G7 and G8 is 8 carbon atoms; in another embodiment, the total length of G7 and G8 is 9 carbon atoms; in another embodiment, the total length of G7 and G8 is 10 carbon atoms; in another embodiment, the total length of G7 and G8 is 11 carbon atoms; in another embodiment, the total length of G7 and G8 is 12 carbon atoms.

[0846] In one embodiment, the total length of G7 and G8 is 6, 7, 8, 9 or 10 carbon atoms. In one embodiment, the total length of G7 and G8 is 6, 7 or 8 carbon atoms.

[0847] In one embodiment, G9 is absent; in another embodiment, G9 is C 1-12 Alkylene; in another embodiment, G9 is C 1-6 Alkylene; in another embodiment, G9 is C 1-5Alkylene; in another embodiment, G9 is C 1-5 linear alkylene; in another embodiment, G9 is -CH2-; in another embodiment, G9 is -(CH2)2-; in another embodiment, G9 is -(CH2)4-; in another embodiment, G9 is -(CH2)5-; in another embodiment, G9 is optionally substituted with 1, 2, 3, 4, 5, or 6 R, wherein R is C 1-10 alkyl; in another embodiment, 1, 2 or 3 methylene groups in G9 are optionally and independently substituted with 1 R; in another embodiment, 1 or 2 methylene groups in G9 are optionally and independently substituted with 1 R; in another embodiment, the methylene group of G9 connected to M2 is not substituted with R.

[0848] In one embodiment, G 10 is absent; in another embodiment, G 10 It is C 1-12 Alkylene; in another embodiment, G 10 It is C 1-10 Alkylene; in another embodiment, G 10 It is C 1-8 Alkylene; in another embodiment, G 10 It is C 1-8 linear alkylene; in another embodiment, G 10 is -(CH2)2-; in another embodiment, G 10 is -(CH2)4-; in another embodiment, G 10 is -(CH2)6-; in another embodiment, G 10 is -(CH2)7-; in another embodiment, G 10 is -(CH2)8-; in another embodiment, G 10 Optionally substituted with 1, 2, 3, 4, 5 or 6 R, wherein R is C 1-10 Alkyl; in another embodiment, G 10 1, 2 or 3 methylene groups in are optionally and independently substituted by 1 R; in another embodiment, G 10 1 or 2 methylene groups in are optionally and independently substituted by 1 R.

[0849] In one embodiment, G9 and G 10 The total length of G9 and G is 4 carbon atoms; in another embodiment, 10 The total length of G9 and G is 5 carbon atoms; in another embodiment, 10 The total length of G9 and G is 6 carbon atoms; in another embodiment, 10The total length of G9 and G is 7 carbon atoms; in another embodiment, 10 The total length of G9 and G is 8 carbon atoms; in another embodiment, 10 The total length of G9 and G is 9 carbon atoms; in another embodiment, 10 The total length of G9 and G is 10 carbon atoms; in another embodiment, 10 The total length of G9 and G is 11 carbon atoms; in another embodiment, 10 The total length is 12 carbon atoms.

[0850] In one embodiment, G9 and G 10 The total length of G9 is 6, 7, 8, 9 or 10 carbon atoms. 10 The total length of the molecule is 6, 7 or 8 carbon atoms.

[0851] In one embodiment, R s is H. In one embodiment, R s It is C 1-10 In one embodiment, R s It is C 1-6 In one embodiment, R s In one embodiment, R s In one embodiment, R s In one embodiment, R s In one embodiment, R s In one embodiment, R s In one embodiment, R s Yes-L d -OR d In one embodiment, R s Yes-L d -NR d R' d In one embodiment, R s Yes-OR d In one embodiment, R s Yes-NR d R' d In one embodiment, R s Yes -CH2-OR d In one embodiment, R s Yes-CH2-NR d R' d .

[0852] In one embodiment, R s' is H. In one embodiment, R s 'It's C 1-10 In one embodiment, R s 'It's C 1-6 In one embodiment, R s ' is methyl. In one embodiment, R s ' is ethyl. In one embodiment, R s ' is a C3 alkyl group. In one embodiment, R s ' is C4 alkyl. In one embodiment, R s ' is a C5 alkyl group. In one embodiment, R s ' is a C6 alkyl group. In one embodiment, R s 'Yes-L d -OR d In one embodiment, R s 'Yes-L d -NR d R' d In one embodiment, R s 'Yes-OR d In one embodiment, R s 'Yes-NR d R' d In one embodiment, R s ' is -CH2-OR d In one embodiment, R s ' is -CH2-NR d R' d .

[0853] Each instance of R and R' is independently H, C 1-20 Alkyl, L a -OR a 、-L a -SR a or -L a -NR a R' a , or R and R' together with the carbon atom to which they are attached form C 3-8 cycloalkylene;

[0854] In one embodiment, R is H. In one embodiment, R is C 1-20 In one embodiment, R is C 1-14 In one embodiment, R is C 1-8 In one embodiment, R is C 1-6 In one embodiment, R is -L a -OR aIn one embodiment, R is -L a -NR a R' a In one embodiment, R is -OR a In one embodiment, R is -NR a R' a In one embodiment, R is -CH2-OR a In one embodiment, R is -CH2-NR a R' a .

[0855] In one embodiment, R' is H. In one embodiment, R' is C 1-20 In one embodiment, R' is C 1-14 In one embodiment, R' is C 1-8 In one embodiment, R' is C 1-6 In one embodiment, R' is -L a -OR a In one embodiment, R' is -L a -NR a R' a In one embodiment, R' is -OR a In one embodiment, R' is -NR a R' a In one embodiment, R' is -CH2-OR a In one embodiment, R' is -CH2-NR a R' a .

[0856] In one embodiment, R and R' together with the carbon atoms to which they are attached form C 3-8 In one embodiment, R and R' together with the carbon atom to which they are attached form C 3-6 In one embodiment, R and R' together with the carbon atoms to which they are attached form C3 cycloalkylene, C4 cycloalkylene, C5 cycloalkylene or C6 cycloalkylene,

[0857] In one embodiment, R" is H. In one embodiment, R" is C 1-14 In one embodiment, R" is C 1-8 In one embodiment, R" is C 1-6Alkyl. In one embodiment, R" is methyl. In one embodiment, R" is ethyl. In one embodiment, R" is C3 alkyl. In one embodiment, R" is C4 alkyl. In one embodiment, R" is C5 alkyl. In one embodiment, R" is C6 alkyl.

[0858] In one embodiment, L d In one embodiment, L d It is C 1-10 In one embodiment, L d It is C 1-6 In one embodiment, L d In one embodiment, L d In one embodiment, L d is C3 alkylene. In one embodiment, L d In one embodiment, L d In one embodiment, L d It is a C6 alkylene group.

[0859] In one embodiment, R c is H. In one embodiment, R c It is C 1-8 In one embodiment, R c It is C 1-6 In one embodiment, R c In one embodiment, R c In one embodiment, R c In one embodiment, R c In one embodiment, R c In one embodiment, R c In one embodiment, R c In one embodiment, R c It is a C8 alkyl group.

[0860] In one embodiment, R' c Is H. In one embodiment, R' c It is C 1-8 In one embodiment, R' c It is C 1-6 In one embodiment, R' c In one embodiment, R' c In one embodiment, R' cIs a C3 alkyl group. In one embodiment, R' c In one embodiment, R' c is a C5 alkyl group. In one embodiment, R' c In one embodiment, R' is a C6 alkyl group. In one embodiment, R' is a C7 alkyl group. c It is a C8 alkyl group.

[0861] In one embodiment, R d is H. In one embodiment, R d It is C 1-10 In one embodiment, R d It is C 1-8 In one embodiment, R d It is C 1-6 In one embodiment, R d In one embodiment, R d In one embodiment, R d In one embodiment, R d In one embodiment, R d In one embodiment, R d In one embodiment, R d In one embodiment, R d In one embodiment, R d In one embodiment, R d It is C 10 alkyl.

[0862] In one embodiment, R' d Is H. In one embodiment, R' d It is C 1-10 In one embodiment, R' d It is C 1-8 In one embodiment, R' d It is C 1-6 In one embodiment, R' d In one embodiment, R' d In one embodiment, R' d Is a C3 alkyl group. In one embodiment, R' d In one embodiment, R' d is a C5 alkyl group. In one embodiment, R' d is a C6 alkyl group. In one embodiment, R' dis a C7 alkyl group. In one embodiment, R' d In one embodiment, R' d In one embodiment, R' d It is C 10 alkyl.

[0863] In one embodiment, a' is 0. In one embodiment, a' is 1. In one embodiment, a' is 2. In one embodiment, a' is 3. In one embodiment, a' is 4. In one embodiment, a' is 5.

[0864] In one embodiment, b is 0. In one embodiment, b is 1. In one embodiment, b is 2. In one embodiment, b is 3. In one embodiment, b is 4. In one embodiment, b is 5.

[0865] In one embodiment, g is 0. In one embodiment, g is 1. In one embodiment, g is 2. In one embodiment, g is 3. In one embodiment, g is 4. In one embodiment, g is 5.

[0866] In one embodiment, a' is 2 and b is 2. In one embodiment, a' is 0 and b is 2. In one embodiment, a' is 2 and b is 0. In one embodiment, a' is 1 and b is 2. In one embodiment, a' is 2 and b is 1.

[0867] In one embodiment, a' + g is equal to 2. In one embodiment, a' + g is equal to 3. In one embodiment, a' + g is equal to 0. In one embodiment, a' + g is equal to 1. In one embodiment, a' + g is equal to 4. In one embodiment, a' + g is equal to 5.

[0868] In one embodiment, c is 0. In one embodiment, c is 1. In one embodiment, c is 2. In one embodiment, c is 3. In one embodiment, c is 4. In one embodiment, c is 5. In one embodiment, c is 6. In one embodiment, c is 7.

[0869] In one embodiment, d is 0. In one embodiment, d is 1. In one embodiment, d is 2. In one embodiment, d is 3. In one embodiment, d is 4. In one embodiment, d is 5. In one embodiment, d is 6. In one embodiment, d is 7.

[0870] In one embodiment, e is 0. In one embodiment, e is 1. In one embodiment, e is 2. In one embodiment, e is 3. In one embodiment, e is 4. In one embodiment, e is 5. In one embodiment, e is 6. In one embodiment, e is 7.

[0871] In one embodiment, f is 0. In one embodiment, f is 1. In one embodiment, f is 2. In one embodiment, f is 3. In one embodiment, f is 4. In one embodiment, f is 5. In one embodiment, f is 6. In one embodiment, f is 7.

[0872] In one embodiment, c + d is equal to 2. In one embodiment, c + d is equal to 3. In one embodiment, c + d is equal to 0. In one embodiment, c + d is equal to 1. In one embodiment, c + d is equal to 4. In one embodiment, c + d is equal to 5. In one embodiment, c + d is equal to 6. In one embodiment, c + d is equal to 7. In one embodiment, c + d is equal to 8. In one embodiment, c + d is equal to 9.

[0873] In one embodiment, e + f equals 2. In one embodiment, e + f equals 3. In one embodiment, e + f equals 0. In one embodiment, e + f equals 1. In one embodiment, e + f equals 4. In one embodiment, e + f equals 5. In one embodiment, e + f equals 6. In one embodiment, e + f equals 7. In one embodiment, e + f equals 8. In one embodiment, e + f equals 9.

[0874] In one embodiment, The total length of is 4, 5, 6, 7, 8 or 9 carbon atoms.

[0875] In one embodiment, Independently selected from: -(CH2)3-C(CH3)2-, -(CH2)4-C(CH3)2-, -(CH2)5-C(CH3)2-, -(CH2)6-C(CH3)2-, -(CH2)7-C(CH3)2-, -(CH2)8-C(CH3)2-, -(CH2)3-CH=CH-C(CH3)2-, -(CH2)3-C≡CC(CH3)2-, -(CH2)4-C(CH3)2-CH2-, -(CH 2) 3-C(CH3)2-(CH2)2-, -(CH2)2-C(CH3)2-(CH2)3-, -(CH2)2-CH=CH-C(CH3)2-CH2-, -(CH2)2-C(CH3)2-C≡C-CH2-, -(CH2)2-C(CH3)2-CH=CH-CH2-, -(CH2)2-C(CH3)2-CH=CH-CH2-, -(CH2)2-C≡CC(CH3)2-CH2- and -(CH2)3-C(CH3)2-C≡C-; in one embodiment, are independently -(CH2)4-C(CH3)2-, -(CH2)5-C(CH3)2-, or -(CH2)6-C(CH3)2-; in one embodiment, In one embodiment, is -(CH2)6-C(CH3)2-, and In one embodiment, is -(CH2)7-, and It is -(CH2)7-.

[0876] In one embodiment, -G7-L1-G8-H or -G9-L2-G 10 -H is independently selected from: -(CH2)5CH3, -(CH2)6CH3, -(CH2)7CH3, -(CH2)8CH3, -(CH2)9CH3, -(CH2) 10 CH3, -(CH2) 11 CH3, -CH2-C≡C-(CH2)5CH3, -CH2-C≡C-(CH2)6CH3, -(CH2)2-C≡C-(CH2)5CH3, -(CH2)4-C≡C-(CH2)3CH3, -CH2-CH=CH-( CH2)5CH3, -CH2-CH=CH-(CH2)6CH3, -(CH2)2-CH=CH-(CH2)5CH3, -(CH2)4-CH=CH-(CH2)3CH3, -(CH2)5-CH=CH-CH2CH3,

[0877] In one embodiment, -G7-L1-G8-H or -G9-L2-G 10 -H is independently selected from the following groups: -(CH2)5CH3, -(CH2)6CH3, -(CH2)7CH3, -(CH2)8CH3, -(CH2)9CH3, -(CH2) 10 CH3, -(CH2) 11 CH3, -CH2-C≡C-(CH2)5CH3, -CH2-C≡C-(CH2)6CH3, -(CH2)2-C≡C-(CH2)5CH3, -(CH2)4-C≡C-(CH2)3CH3, -CH2-CH=CH-( CH2)5CH3, -CH2-CH=CH-(CH2)6CH3, -(CH2)2-CH=CH-(CH2)5CH3, -(CH2)4-CH=CH-(CH2)3CH3, -(CH2)5-CH=CH-CH2CH3,

[0878] In one embodiment, -G7-L1-G8-H or -G9-L2-G 10 -H is In one embodiment, -G7-L1-G8-H or -G9-L2-G 10 -H is -(CH2)8CH3. In one embodiment, -G7-L1-G8-H and -G9-L2-G 10 One of the -H is And another is -(CH2)8CH3.

[0879] In one embodiment, the ionizable lipid is a compound of formula (IX):

[0880] or a stereoisomer, a mixture of stereoisomers or a pharmaceutically acceptable salt thereof, wherein

[0881] G1 and G2 are each independently a bond, C 1-13 Straight chain alkylene, C 2-13 Straight chain alkenylene or C 2-13 Straight chain alkynylene, wherein the alkylene, alkenylene and alkynylene groups are optionally substituted by one or more R G1 replace;

[0882] The total length of G1 and G2 is 3, 4, 5, 6, 7, 8, 9, 10, 11, 12 or 13 carbon atoms;

[0883] R G1In each case, independently H, C 1-14 Alkyl, -L a -OR a 、-L a -SR a or -L a -NR a R' a ;

[0884] G3 is C 4-14 Straight chain alkylene, C 4-14 Straight chain alkenylene or C 4-14 Straight chain alkynylene, wherein the alkylene, alkenylene and alkynylene groups are optionally substituted by one or more R G3 replace;

[0885] R G3 In each case, independently H, -L a -OR a 、-L a -SR a or -L a -NR a R' a ;

[0886] L a independently a bond or C 1-14 alkylene;

[0887] R a and R' a Each independently is H, C 1-14 Alkyl, C 3-14 Cycloalkyl or 3-14 membered heterocyclic group;

[0888] G4 is the key, C 1-6 Alkylene, C 2-6 Alkenylene or C 2-6 Alkyne, wherein the alkylene, alkenylene and alkynylene groups are optionally substituted by one or more R G4 replace;

[0889] R G4 In each case, independently H, C 1-6 Alkyl, -L b -OR b 、-L b -SR b or -L b -NR b R' b ;

[0890] L b independently a bond or C 1-6 alkylene;

[0891] R b and R' b Independently H, C 1-6 Alkyl, C 3-10 Cycloalkyl or 3-10 membered heterocyclic group;

[0892] Or two R G4 Together with the carbon atom to which they are attached, they form C 3-14 Cycloalkylene or 3-14 membered heterocyclylene, wherein the cycloalkylene and heterocyclylene are optionally substituted by one or more R 4g replace;

[0893] R 4g is independently H, halogen, cyano, C 1-8 Alkyl, C 1-8 Halogenated alkyl, -L e -OR e 、-L e -SR e or -L e -NR e R' e ;

[0894] L e independently a bond or C 1-8 alkylene;

[0895] R e and R' e Independently H, C 1-8 Alkyl, C 3-14 Cycloalkyl or 3-14 membered heterocyclic group;

[0896] M1 and M2 are independently -C(O)O-, -OC(O)-, -O-, -SC(O)O-, -OC(O)NR-, -NRC(O)NR-, -OC(O)S-, -OC(O)O-, -NRC(O)O-, -SC(O)-, -C(O )S-, -NR-, -C(O)NR-, -NRC(O)-, -NRC(O)S-, -SC(O)NR-, -C(O)-, -OC(S)-, -C(S)O-, -OC(S)NR-, -NRC(S)O-, -SS- or -S(O) 0-2 -;

[0897] Q is a bond, -C(O)O-, -O-, -SC(O)O-, -OC(O)NR f -、-NR f C(O)NR f -、-OC(O)S-、-OC(O)O-、-NR fC(O)O-, -OC(O)-, -SC(O)-, -C(O)S-, -NR f -、-C(O)NR f -、-NR f C(O)-、-NR f C(O)S-、-SC(O)NR f -, -C(O)-, -OC(S)-, -C(S)O-, -OC(S)NR f -、-NR f C(S)O-, -SS-, -S(O) 0-2 -, phenylene or pyridinylene, wherein the phenylene or pyridinylene is optionally substituted with one or more R*;

[0898] R* is independently H, halogen, cyano, C 1-10 Alkyl, C 1-10 Halogenated alkyl, -L f -OR f 、-L f -SR f or -L f -NR f R' f ;

[0899] L f independently a bond or C 1-8 alkylene;

[0900] R f and R' f Independently H, C 1-10 Alkyl, C 3-14 Cycloalkyl or 3-14 membered heterocyclic group;

[0901] R1 and R2 are independently C 4-20 Alkyl, C 4-20 Alkenyl or C 4-20 Alkynyl, which is optionally substituted by one or more R 1s substituted, and wherein one or more methylene units are optionally and independently replaced by -NR'-;

[0902] R 1s Independently H, C 1-20 Alkyl, -L c -OR c 、-L c -SR c or -L c -NR c R' c ;

[0903] R and R' are independently H or C 1-20 alkyl;

[0904] L c independently a bond or C 1-20 alkylene;

[0905] R c and R' c Independently H, C 1-20 Alkyl, C 3-14 Cycloalkyl or 3-14 membered heterocyclic group;

[0906] R3 is -CN, -OR g 、-C(O)R g 、-OC(O)R g 、-NR”C(O)R g 、-NR g R' g 、-NR”C(O)NR g R' g 、-NR”C(O)R g 、-NR”S(O)2R g 、-OC(O)NR g R' g 、-NR”C(O)OR g 、-N(OR g )C(O)R g 、-N(OR g )S(O)2R g 、-N(OR g )C(O)OR g 、-N(OR g )C(O)R g R' g , 3-14 membered heterocyclyl or 5-14 membered heteroaryl;

[0907] R g and R' g Independently H, C 1-10 Alkyl, C 3-10 Cycloalkyl and 3-10 membered heterocyclic groups;

[0908] R" is independently H or C 1-6 alkyl;

[0909] R5 and R6 are independently C 1-8 Alkyl, wherein the alkyl group is optionally substituted by one or more R 4s replace;

[0910] Or R5 and R6 together with the carbon atom to which they are attached form C 3-14 Cycloalkylene or 3-14 membered heterocyclylene, wherein the cycloalkylene and heterocyclylene are optionally substituted by one or more R 4sreplace;

[0911] R 4s are independently H, halogen, cyano, C 1-8 Alkyl, C 1-8 Halogenated alkyl, -L d -OR d 、-L d -SR d or -L d -NR d R' d ;

[0912] L d independently a bond or C 1-8 alkylene;

[0913] R d and R' d Independently H, C 1-8 Alkyl, C 3-14 Cycloalkyl or 3-14 membered heterocyclic group.

[0914] In one embodiment, G1 and G2 are independently a bond, C 1-9 Straight chain alkylene, C 2-9 Straight chain alkenylene or C 2-9 Straight chain alkynylene.

[0915] In one embodiment, G1 is C 1-6 Straight chain alkylene, C 2-6 Straight chain alkenylene or C 2-6 In one embodiment, G1 is C 1-6 In one embodiment, G1 is C 2-6 Straight chain alkylene.

[0916] In one embodiment, G2 is a bond, C 1-6 Straight chain alkylene, C 2-6 Straight chain alkenylene or C 2-6 In one embodiment, G2 is a bond or C 1-6 In one embodiment, G2 is a bond or C 1-4 Straight chain alkylene.

[0917] In one embodiment, G1 and G2 have a total length of 3, 4, 5, 6, 7, 8 or 9 carbon atoms. In one embodiment, G1 and G2 have a total length of 4, 5 or 6 carbon atoms. In one embodiment, G1 and G2 have a total length of 5 or 6 carbon atoms. In one embodiment, G1 and G2 have a total length of 5, 6 or 7 carbon atoms. In one embodiment, G1 and G2 have a total length of 6 or 7 carbon atoms. In one embodiment, G1 and G2 are optionally replaced by 1, 2, 3 or 4 R G1 replace.

[0918] In one embodiment, at each occurrence of R G1 are independently H or C 1-10 In one embodiment, at each occurrence of R G1 are independently H or C 1-6 alkyl.

[0919] In one embodiment, G3 is C 4-10 Straight chain alkylene, C 4-10 Straight chain alkenylene or C 4-10 In one embodiment, G3 is C 4-9 In one embodiment, G3 is C 5-8 In one embodiment, G3 is optionally replaced by 1, 2, 3 or 4 R G3 replace.

[0920] In one embodiment, L a independently a bond or C 1-10 In one embodiment, L a independently a bond or C 1-6 Alkylene.

[0921] In one embodiment, R a and R' a Each independently is H, C 1-10 Alkyl, C 3-10 Cycloalkyl or 3-10 membered heterocyclyl. In one embodiment, R a and R' a are each independently H or C 1-6 alkyl.

[0922] In one embodiment, G4 is C 1-4 Alkylene, C 2-4 Alkenylene or C 2-4 In one embodiment, G4 is C 2-4 In one embodiment, G4 is C 2-3In one embodiment, G4 is optionally replaced by 1, 2, 3 or 4 R G4 replace.

[0923] In one embodiment, R G4 are independently H or C 1-6 In one embodiment, R G4 Independently C 1-4 alkyl.

[0924] In one embodiment, both R G4 Together with the same carbon atom to which they are attached, they form C 3-10 In one embodiment, two R G4 Together with the same carbon atom to which they are attached, they form C 3-7 In one embodiment, the cycloalkylene and heterocyclylene groups are optionally substituted by 1, 2 or 3 R 4g replace;

[0925] In one embodiment, R 4g are independently H, halogen, cyano, C 1-6 Alkyl or C 1-6 Halogenated alkyl.

[0926] In one embodiment, L b independently a bond or C 1-4 Alkylene.

[0927] In one embodiment, R b and R' b Independently H, C 1-6 Alkyl, C 3-7 Cycloalkyl or 3- to 7-membered heterocyclyl. In one embodiment, R b and R' b are independently H or C 1-6 In one embodiment, R b and R' b are independently H or C 1-4 alkyl.

[0928] In one embodiment, M1 and M2 are independently -C(O)O-, -OC(O)-, -OC(O)O-, -SC(O)-, -C(O)S-, -C(O)NR-, or -NRC(O)-. In one embodiment, M1 and M2 are independently -C(O)O-, -OC(O)O-, -OC(O)-, -SC(O)-, or -C(O)S-. In one embodiment, M1 and M2 are independently -C(O)O-, -OC(O)-, -SC(O)-, or -C(O)S-. In one embodiment, M1 and M2 are independently -C(O)O- or -OC(O)-.

[0929] In one embodiment, R1 and R2 are independently C 6-14 Alkyl, C 6-14 Alkenyl or C 6-14 In one embodiment, R1 and R2 are independently C 6-14 In one embodiment, R1 and R2 are independently C 7-12 In one embodiment, R1 and R2 are independently C 8-12 In one embodiment, R1 and R2 are independently C 7-12 Alkyl, C 7-12 Alkenyl or C 7-12 Alkynyl.

[0930] In one embodiment, R1 and R2 are optionally replaced by 1, 2, 3 or 4 R 1s In one embodiment, R1 and R2 are optionally replaced by one R 1s replace.

[0931] In one embodiment, R1 and R2 are independently -(CH2)5CH3, -(CH2)6CH3, -(CH2)7CH3, -(CH2)8CH3, -(CH2)9CH3, -(CH2) 10 CH3, -(CH2) 11 CH3, -CH2-C≡C-(CH2)5CH3, -CH2-C≡C-(CH2)6CH3, -(CH2)2-C≡C-(CH2)5CH3, -(CH2)2-C≡C-(CH2)4CH3, -(CH2)3-C≡C-(CH2)3CH3, -(CH2)4-C≡C- (CH2)3CH3, -CH2-CH=CH-(CH2)5CH3, -CH2-CH=CH-(CH2)6CH3, -(CH2)2-CH=CH-(CH2)5CH3, -(CH2)4-CH=CH-(CH2)3CH3, -(CH2)5-CH=CH-CH2CH3,

[0932] In one embodiment, R 1s Independently H, C 1-14 Alkyl, -L c -OR c or -L c -NR c R' c In one embodiment, R 1s are independently H or C 1-14 In one embodiment, R 1s are independently H or C 1-10 In one embodiment, R 1s are independently H or C 1-9 In one embodiment, R 1s are independently H or C 1-6 In one embodiment, R 1s are independently H or C 1-4 alkyl.

[0933] In one embodiment, R and R' are independently H or C 1-20 In one embodiment, R and R' are independently H or C 1-14 In one embodiment, R and R' are independently H or C 1-9 In one embodiment, R and R' are independently H or C 1-6 In one embodiment, R is H.

[0934] In one embodiment, L c independently a bond or C 1-14 In one embodiment, L c independently a bond or C 1-10 In one embodiment, L c independently a bond or C 1-6 Alkylene.

[0935] In one embodiment, R c and R' c are independently H or C 1-14 In one embodiment, R c and R' c are independently H or C 1-10 In one embodiment, R c and R' c are independently H or C 1-6 alkyl.

[0936] In one embodiment, R3 is cyano, -ORg or -NR g R' g In one embodiment, R3 is -OR g or -NR g R' g In one embodiment, R3 is -OR g。 In one embodiment, R3 is -OH.

[0937] In one embodiment, R3 is -OH or -N(CH3)2.

[0938] In one embodiment, R g and R' g Independently H, C 1-6 Alkyl, C 3-7 In one embodiment, R g and R' g are independently H or C 1-6 In one embodiment, R g and R' g are independently H or C 1-4 In one embodiment, R g and R' g are independently H or -CH3.

[0939] In one embodiment, R5 and R6 are independently C 1-6 In one embodiment, R5 and R6 are independently C 1-3 In one embodiment, R5 and R6 are independently -CH3. In one embodiment, R5 and R6 are each independently and optionally replaced by 1, 2 or 3 R 4s replace.

[0940] In one embodiment, R5 and R6 together with the carbon atom to which they are attached form C 3-10 In one embodiment, the cycloalkylene or heterocyclylene is optionally substituted by 1, 2 or 3 R 4s replace.

[0941] In one embodiment, R 4s are independently H, halogen, cyano, C 1-6 Alkyl, C 1-6 Halogenated alkyl, -L d -OR d 、-L d -SR d or -L d -NR d R' dIn one embodiment, R 4s are independently H, halogen, cyano, C 1-6 Alkyl or C 1-6 In one embodiment, R 4s Independently H, C 1-3 Alkyl or C 1-3 Halogenated alkyl.

[0942] In one embodiment, L d independently a bond or C 1-6 In one embodiment, L d independently a bond or C 1-3 Alkylene.

[0943] In one embodiment, R d and R' d Independently H, C 1-6 Alkyl, C 3-10 Cycloalkyl or 3-10 membered heterocyclyl. In one embodiment, R d and R' d are independently H or C 1-6 In one embodiment, R d and R' d are independently H or C 1-6 alkyl.

[0944] In one embodiment, L e independently a bond or C 1-6 In one embodiment, L e independently a bond or C 1-4 Alkylene.

[0945] In one embodiment, R e and R' e Independently H, C 1-6 Alkyl, C 3-10 Cycloalkyl or 3-10 membered heterocyclyl. In one embodiment, R e and R' e are independently H or C 1-6 In one embodiment, R e and R' e are independently H or C 1-4 alkyl.

[0946] In one embodiment, L f independently a bond or C 1-6 In one embodiment, L f independently a bond or C 1-4 Alkylene.

[0947] In one embodiment, R f and R' f Independently H, C 1-6 Alkyl, C 3-10 Cycloalkyl or 3-10 membered heterocyclyl. In one embodiment, R f and R' f are independently H or C 1-6 In one embodiment, R f and R' f are independently H or C 1-4 alkyl.

[0948] In one embodiment, the ionizable lipid is a compound of formula (X):

[0949] or a stereoisomer, a mixture of stereoisomers or a pharmaceutically acceptable salt thereof, wherein

[0950] a is 1, 2, 3, 4, 5, or 6;

[0951] b is 4, 5, 6, 7, 8, 9, or 10;

[0952] c is 1, 2, 3, 4, 5, or 6;

[0953] d is 0, 1, 2, 3, or 4;

[0954] c+d is 3, 4, 5, 6, 7, 8 or 9.

[0955] In one embodiment, the M1 or M2 and R on R1 or R2 1s The substitution sites are separated by 0-10 carbon atoms. In one embodiment, the M1 or M2 and R1 or R2 are separated by 0-10 carbon atoms. 1s The substitution sites are separated by 0-6 carbon atoms. In one embodiment, the M1 or M2 and R1 or R2 are separated by 0-6 carbon atoms. 1s The substitution sites are separated by 0-4 carbon atoms. In one embodiment, the M1 or M2 and R1 or R2 are separated by 0-4 carbon atoms. 1s The substitution sites are separated by 0-2 carbon atoms. In one embodiment, the M1 or M2 and R1 or R2 are separated by 0-2 carbon atoms. 1s The substitution sites are separated by 0 carbon atoms.

[0956] In one embodiment, R2 is In one embodiment, R2 is In one embodiment, R2 is In one embodiment, R2 is

[0957] In one embodiment, R1 is

[0958] In a preferred embodiment, the ionizable lipid is:

[0959] or a stereoisomer, a mixture of stereoisomers or a pharmaceutically acceptable salt thereof.

[0960] Descriptions related to ionizable lipids, including their preparation processes, can be found in CN 115850104 A, the entire contents of which are incorporated herein by reference.

[0961] In one embodiment, the ionizable lipid is DODAP, DODMA, DLinDMA, DLin-KC2-DMA, DLin-MC3-DMA, SM-102, or ALC-0315, or a combination thereof, or a stereoisomer, mixture of stereoisomers, or pharmaceutically acceptable salt thereof.

[0962] In one embodiment, the ionizable lipid is compound 46 of the formula:

[0963] or a stereoisomer, a mixture of stereoisomers or a pharmaceutically acceptable salt thereof.

[0964] In one embodiment, the ionizable lipid is Compound 132,

[0965] or a stereoisomer, a mixture of stereoisomers or a pharmaceutically acceptable salt thereof.

[0966] In one embodiment, the ionizable lipid is:

[0967] or a stereoisomer, a mixture of stereoisomers or a pharmaceutically acceptable salt thereof.

[0968] In one embodiment, the molar percentage of the ionizable lipid to the total lipid in the lipid nanoparticle is 35-70 mol%, more preferably 40-60 mol%, such as 35 mol%, 40 mol%, 45 mol%, 50 mol%, 55 mol% or 60 mol%.

[0969] 5.2.3. Phospholipids

[0970] As used herein, phospholipid refers to an amphipathic lipid with a negatively charged phosphate group and one or more hydrophobic moieties. Phospholipids can be assembled into one or more lipid bilayers. Phospholipids are composed of two fatty acid tails (hydrophobic moieties) and a phosphate group head connected by glycerol, and can be natural or synthetic. The limiting examples of phospholipids include soybean lecithin, egg lecithin, phosphatidylglycerol, phosphatidylinositol, phosphatidylethanolamine, phosphatidic acid, sphingomyelin, diphosphatidylglycerol, phosphatidylserine, phosphatidylcholine, dimyristoylphosphatidylcholine, dimyristoylphosphatidylglycerol, distearoylphosphatidylglycerol, dipalmitoylphosphatidylcholine and hydrogenated or partially hydrogenated phosphatidylcholine. In some embodiments, the phospholipid is phosphatidic acid (DMPA, DPPA, DSPA), phosphatidylcholine (DDPC, DLPC, DMPC, DPPC, DSPC, DOPC, POPC, DEPC), phosphatidylglycerol (DMPG, DPPG, DSPG, POPG), phosphatidylethanolamine (DMPE, DPPE, DSPE, DOPE), or phosphatidylserine (DOPS). The phospholipid can be a natural phospholipid, a non-natural phospholipid, or a synthetic phospholipid. The phospholipid can be modified or functionalized by various chemical reactions, such as attachment to an alkyne to conjugate with an azide group. The phospholipid can be functionalized for targeting or imaging purposes.

[0971] In one embodiment, the lipid nanoparticles comprise phospholipids. In one embodiment, the phospholipid is distearoylphosphatidylcholine (DSPC). In one embodiment, the phospholipid is dioleoylphosphatidylethanolamine (DOPE). In one embodiment, the phospholipid is dimyristoylphosphatidylcholine (DMPC). In one embodiment, the phospholipid is 1,2-dioleoyl-sn-glycero-3-phosphocholine (DOPC). In one embodiment, the phospholipid is dipalmitoylphosphatidylcholine (DPPC). In one embodiment, the phospholipid is 1-palmitoyl-2-oleoyl-sn-glycero-3-phosphocholine (POPC). In one embodiment, the phospholipid is 1,2-dimyristoyl-sn-glycero-3-phosphoethanolamine (DMPE). In one embodiment, the phospholipid is 1,2-dipalmitoyl-sn-glycero-3-phosphoethanolamine (DPPE). In one embodiment, the phospholipid is dipalmitoylphosphatidylcholine (DPPC). In one embodiment, the phospholipid is 2-(9Z-octadecanoyl)-sn-glycero-3-phosphoethanolamine (POPE).

[0972] In one embodiment, the lipid nanoparticles do not comprise phospholipids, or comprise phospholipids in an amount less than about 50 mol%, about 40 mol%, 30 mol%, 20 mol%, 15 mol%, 10 mol% or 5 mol% of the total lipids present in the lipid nanoparticles.

[0973] In one embodiment, the molar percentage of the phospholipids to the total lipids in the lipid nanoparticles is 5-30 mol%, more preferably 5-20 mol%, more preferably 5-15 mol%, for example 5 mol%, 6 mol%, 7 mol%, 8 mol%, 9 mol%, 10 mol%, 11 mol%, 12 mol%, 13 mol%, 14 mol%, 15 mol%, 16 mol%, 17 mol%, 18 mol%, 19 mol% or 20 mol%.

[0974] 5.2.4. Structural lipids

[0975] In one embodiment, the lipid nanoparticles further comprise a structured lipid. Unless otherwise specified, the structured lipid is a neutral lipid other than the steroid compound described in Section 0. In one embodiment, the structured lipid does not comprise a cationic group. In one embodiment, the structured lipid does not comprise a tertiary amine group or a quaternary ammonium group.

[0976] In one embodiment, the structured lipid is a sterol. In one embodiment, the structured lipid is a neutral sterol. In one embodiment, the structured lipid is cholesterol. In certain embodiments, the structured lipid is an analog of cholesterol. In one embodiment, the structured lipid is sitosterol or β-sitosterol. In one embodiment, the structured lipid is coprosterol. In one embodiment, the structured lipid is fucoxosterol. In one embodiment, the structured lipid is brassicasterol. In one embodiment, the structured lipid is ergosterol. In one embodiment, the structured lipid is tomatine. In one embodiment, the structured lipid is ursolic acid. In one embodiment, the structured lipid is α-tocopherol. In one embodiment, the structured lipid is stigmasterol. In one embodiment, the structured lipid is avenasterol. In one embodiment, the structured lipid is campesterol. In one embodiment, the structured lipid is solanine.

[0977] In one embodiment, the molar percentage of the structural lipids in the lipid nanoparticles is 10-80 mol%, more preferably 15-70 mol%, more preferably 30-50 mol%, more preferably 30-40 mol%, for example 30 mol%, 35 mol%, 40 mol%, 45 mol%, 50 mol%, 55 mol% or 60 mol%.

[0978] 5.2.5. Polymer-conjugated lipids

[0979] In one embodiment, lipid nanoparticles include polymer-conjugated lipids. In one embodiment, polymer-conjugated lipids are PEGylated lipids (PEG lipids). In one embodiment, PEG lipids are diglycerides, which also include PEG chains connected to glycerol groups. In one embodiment, PEG lipids are compounds containing one or more C6-C24 long-chain alkyl or alkenyl groups or C6-C24 fatty acid groups linked to linker groups with PEG chains. Non-limiting examples of PEG lipids include PEG-modified phosphatidylethanolamine and phosphatidic acid, PEG-ceramide-conjugated PEG-modified dialkylamines and PEG-modified 1,2-diacyloxypropane-3-amines, PEG-modified diacylglycerols and dialkylglycerols. In one embodiment, PEG-modified distearoyl phosphatidylethanolamine or PEG-modified dimyristoyl-sn-glycerol. In one embodiment, PEG modifications are measured according to the molecular weight of the PEG component of the lipid. In one embodiment, the molecular weight of the PEGylated lipid is from about 1000Da to about 10,000Da. In one embodiment, the molecular weight of the PEGylated lipid is from about 1000 Da to about 5000 Da. In one embodiment, the molecular weight of the PEGylated lipid is from about 1000 Da to about 2000 Da.

[0980] In one embodiment, the PEGylated lipid is methoxypolyethylene glycoloxy (2000) -N, N-ditetradecyl acetamide (ALC-0159). In one embodiment, the PEGylated lipid is 1,2-dimyristoyl-rac-glycero-3-methoxypolyethylene glycol-2000 (DMG-PEG2000). In one embodiment, the PEGylated lipid is 1,2-dimyristoyl-sn-glycero-3-phosphoethanolamine-polyethylene glycol 1000 (DMPE-PEG1000). In one embodiment, the PEGylated lipid is 1,2-dipalmitoyl-sn-glycero-3-phosphoethanolamine-polyethylene glycol 1000 (DPPE-PEG1000). In one embodiment, the PEGylated lipid is 1,2-distearoyl-sn-glycero-3-phosphoethanolamine-polyethylene glycol 1000 (DSPE-PEG1000). In one embodiment, the PEGylated lipid is 1,2-dioleoyl-sn-glycero-3-phosphoethanolamine-polyethylene glycol 1000 (DOPE-PEG1000).

[0981] In one embodiment, the PEGylated lipid is 1,2-dimyristoyl-rac-glyceryl-3-methoxypolyethylene glycol-2000 (ceramide-PEG2000). In one embodiment, the PEGylated lipid is 1,2-dimyristoyl-sn-glyceryl-3-phosphoethanolamine-polyethylene glycol 2000 (DMPE-PEG2000). In one embodiment, the PEGylated lipid is 1,2-dipalmitoyl-sn-glyceryl-3-phosphoethanolamine-polyethylene glycol 2000 (DPPE-PEG2000). In one embodiment, the PEGylated lipid is 1,2-distearoyl-sn-glyceryl-3-phosphoethanolamine-polyethylene glycol 2000 (DSPE-PEG2000). In one embodiment, the PEGylated lipid is 1,2-distearoyl-sn-glycero-3-phosphoethanolamine-N-(polyethylene glycol 2000)-mannose (DSPE-PEG2000-mannose). In one embodiment, the PEGylated lipid is 1,2-dimyristoyl-rac-glycero-3-methoxypolyethylene glycol-5000 (ceramide-PEG5000). In one embodiment, the PEGylated lipid is 1,2-distearoyl-sn-glycero-3-phosphoethanolamine-polyethylene glycol 5000 (DSPE-PEG5000). In one embodiment, the PEGylated lipid is 1,2-distearoyl-sn-glycero-3-phosphoethanolamine-N-[amino(polyethylene glycol)-2000] (DSPE-PEG2000 amine).

[0982] In a preferred embodiment, the molar percentage of the polymer-conjugated lipid to the total lipid in the lipid nanoparticles is 0.25-10 mol%, more preferably 0.5-5 mol%, more preferably 0.5-3 mol%, such as 0.5 mol%, 0.6 mol%, 0.7 mol%, 0.8 mol%, 0.9 mol%, 1.0 mol%, 1.1 mol%, 1.2 mol%, 1.3 mol%, 1.4 mol%, 1.5 mol%, 1.6 mol%, 1.7 mol%. , 1.8mol%, 1.9mol%, 2.0mol%, 2.1mol%, 2.2mol%, 2.3mol%, 2.4mol%, 2.5mol%, 2.6mol%, 2.7mol%, 2.8mol%, 2.9 mol%, 3.0 mol%, 3.1 mol%, 3.2 mol%, 3.3 mol%, 3.4 mol%, 3.5 mol%, 3.6 mol%, 3.7 mol%, 3.8 mol%, 3.9 mol%, 4.0 mol%.

[0983] 5.2.6. Payload

[0984] In one embodiment, the lipid nanoparticle comprises a payload.

[0985] In one embodiment, the payload is a small molecule.

[0986] In one embodiment, the payload is a nucleic acid.

[0987] In one embodiment, the payload is an anticancer agent, an antifungal agent, a psychiatric agent (e.g., an analgesic), a consciousness-altering agent (e.g., an anesthetic or hypnotic), a nonsteroidal anti-inflammatory drug (NSAIDS), an anthelmintic, an anti-acne agent, an anti-angina agent, an anti-arrhythmic agent, an anti-asthma agent, an antibacterial agent, an anti-benign prostatic hyperplasia agent, an anticoagulant, an antidepressant, an antidiabetic agent, an antiemetic agent, an antiepileptic agent, an antigout agent, an antihypertensive agent, an anti-inflammatory agent, an antimalarial agent, an anti-migraine agent, an antimuscarinic agent, an antitumor agent, an antiobesity agent, an antiosteoporosis agent, an antiparkinsonian agent, an antiproliferative agent, an antiprotozoal agent, an antithyroid agent, an antitussive agent, an anti-urinary incontinence agent, an antiviral agent, an anxiolytic agent, an appetite suppressant, a beta-blocker, an agent that affects or controls cardiac muscle contraction (cardiac inotropic agents), chemotherapy drugs, cognitive enhancers, birth control pills, corticosteroids, Cox-2 inhibitors, diuretics, erectile dysfunction improvers, expectorants, gastrointestinal preparations, histamine receptor antagonists, immunosuppressants, keratolytics, lipid-regulating agents, leukotriene inhibitors, macrolides, muscle relaxants, neuroleptics, nutritional preparations, opioid analgesics, protease inhibitors, or sedatives.

[0988] In one embodiment, the payload is a protein. In one embodiment, the payload is a peptide. In one embodiment, the payload is an antibody. In one embodiment, the payload is a monoclonal antibody. In one embodiment, the payload is a bispecific antibody.

[0989] In one embodiment, the payload is an antisense oligonucleotide (ASO). In one embodiment, the ASO comprises naturally occurring nucleosides. In one embodiment, the ASO comprises modified nucleosides. In one embodiment, the ASO is capable of regulating the expression of a target gene by hybridizing with a target nucleic acid, particularly a continuous sequence on the target nucleic acid. In one embodiment, the ASO is single-stranded.

[0990] In one embodiment, the payload is deoxyribonucleic acid (DNA). In one embodiment, the payload is plasmid DNA (pDNA). In one embodiment, the payload is double-stranded DNA (dsDNA). In one embodiment, the payload is single-stranded DNA (ssDNA).

[0991] In one embodiment, the payload is ribonucleic acid (RNA). In one embodiment, the payload is RNA interference (RNAi). In one embodiment, the payload is small interfering RNA (siRNA). In one embodiment, the payload is short hairpin RNA (shRNA). In one embodiment, the payload is antisense RNA (aRNA). In one embodiment, the payload is messenger RNA (mRNA). In one embodiment, the payload is modified messenger RNA (mmRNA). In one embodiment, the payload is long non-coding RNA (lncRNA). In one embodiment, the payload is microRNA (miRNA). In one embodiment, the payload is small activating RNA (siRNA). In one embodiment, the payload is multi-coding nucleic acid (MCNA). In one embodiment, the payload is polymer-encoded nucleic acid (PCNA). In one embodiment, the payload is any RNA in a ribozyme.

[0992] In one embodiment, the payload is a clustered regularly interspaced short palindromic repeats (CRISPR)-related nucleic acid. In one embodiment, the payload is a guide RNA (gRNA). In one embodiment, the payload is a CRISPR RNA (crRNA). In one embodiment, the payload comprises a first nucleic acid and a second nucleic acid. In one embodiment, the first nucleic acid is a messenger RNA. In one embodiment, the second nucleic acid is a single guide RNA. In one embodiment, the first nucleic acid is a messenger RNA (mRNA), and the second nucleic acid is a single guide RNA (sgRNA).

[0993] In one embodiment, the ratio of (total nitrogen atoms in the cationic steroid compound and ionizable lipid) to (total phosphorus atoms in the nucleic acid) is from about 1:1 to about 15:1 (N:P ratio). In one embodiment, the N:P ratio is from about 3:1 to about 12:1. In one embodiment, the N:P ratio is from about 4:1 to about 9:1. In one embodiment, the N:P ratio is about 4:1, about 5:1, about 6:1, about 7:1, about 8:1, or about 9:1. In one embodiment, the N:P ratio is about 6:1. In one embodiment, the N:P ratio is 6:1. In one embodiment, the N:P ratio is about 8:1. In one embodiment, the N:P ratio is 8:1. In one embodiment, the N:P ratio is about 4:1. In one embodiment, the N:P ratio is 4:1. In one embodiment, the N:P ratio is about 5.5:1. In one embodiment, the N:P ratio is 5.5:1.

[0994] In one embodiment, when the lipid nanoparticles are administered to a subject, the amount of payload delivered or expressed by the lipid nanoparticles in a non-hepatic tissue of the subject is higher than the amount of payload delivered or expressed by the lipid nanoparticles in the subject's liver. In one embodiment, when the lipid nanoparticles are administered to a subject, the amount of payload delivered or expressed by the lipid nanoparticles in the subject's spleen is higher than the amount of payload delivered or expressed by the lipid nanoparticles in the subject's liver.

[0995] In one embodiment, the amount of payload delivered or expressed in the spleen of a subject is higher than the amount of payload delivered or expressed in the liver of a subject. In one embodiment, the amount of payload delivered or expressed in the spleen of a subject is at least 2 times higher than the amount of payload delivered or expressed in the liver of a subject. In one embodiment, the amount of payload delivered or expressed in the spleen of a subject is at least 5 times higher than the amount of payload delivered or expressed in the liver of a subject. In one embodiment, the amount of payload delivered or expressed in the spleen of a subject is at least 10 times higher than the amount of payload delivered or expressed in the liver of a subject. In one embodiment, the amount of payload delivered or expressed in the spleen of a subject is at least 20 times higher than the amount of payload delivered or expressed in the liver of a subject. In one embodiment, the amount of payload delivered or expressed in the spleen of a subject is at least 40 times higher than the amount of payload delivered or expressed in the liver of a subject. In one embodiment, the amount of payload delivered or expressed in the spleen of a subject is at least 60 times higher than the amount of payload delivered or expressed in the liver of a subject. In one embodiment, the amount of payload delivered or expressed in the spleen of the subject is at least 100-fold greater than the amount of payload delivered or expressed in the liver of the subject.

[0996] In one embodiment, when lipid nanoparticles comprising a nucleic acid are administered to a subject, the amount of protein expressed by the nucleic acid in the subject's spleen is higher than the amount of protein expressed by the nucleic acid in the subject's liver. In one embodiment, when lipid nanoparticles comprising an mRNA are administered to a subject, the amount of protein expressed by the mRNA in the subject's spleen is higher than the amount of protein expressed by the mRNA in the subject's liver.

[0997] In one embodiment, the amount of protein expressed in the spleen of a subject is higher than the amount of protein expressed in the liver of a subject. In one embodiment, the amount of protein expressed in the spleen of a subject is at least 2 times higher than the amount of protein expressed in the liver of a subject. In one embodiment, the amount of protein expressed in the spleen of a subject is at least 5 times higher than the amount of protein expressed in the liver of a subject. In one embodiment, the amount of protein expressed in the spleen of a subject is at least 10 times higher than the amount of protein expressed in the liver of a subject. In one embodiment, the amount of protein expressed in the spleen of a subject is at least 20 times higher than the amount of protein expressed in the liver of a subject. In one embodiment, the amount of protein expressed in the spleen of a subject is at least 40 times higher than the amount of protein expressed in the liver of a subject. In one embodiment, the amount of protein expressed in the spleen of a subject is at least 60 times higher than the amount of protein expressed in the liver of a subject. In one embodiment, the amount of protein expressed in the spleen of a subject is at least 100 times higher than the amount of protein expressed in the liver of a subject.

[0998] 5.3 Lipid Nanoparticle Composition

[0999] In one embodiment, provided herein is a lipid nanoparticle composition comprising a lipid nanoparticle described in Section 0. In one embodiment, a lipid nanoparticle composition described herein comprises a steroid compound described in Section 0 and an ionizable lipid described in Section 0. In one embodiment, a lipid nanoparticle composition described herein comprises a phospholipid described in Section 0. In one embodiment, a population of lipid nanoparticles described herein does not comprise a phospholipid. In one embodiment, a lipid nanoparticle composition described herein comprises a structured lipid described in Section 0. In one embodiment, a lipid nanoparticle composition described herein comprises a polymer-conjugated lipid described in Section 0. In one embodiment, a lipid nanoparticle composition described herein comprises a payload described in Section 0.

[1000] In one embodiment, the lipid nanoparticle composition has a polydispersity index (PDI) of less than 0.2. In one embodiment, the lipid nanoparticle composition has a PDI of less than 0.15. In one embodiment, the lipid nanoparticle composition has a PDI of about 0.01, about 0.02, about 0.03, about 0.04, about 0.05, about 0.06, about 0.07, about 0.08, about 0.09, or about 0.10.

[1001] In one embodiment, the apparent pKa of the lipid nanoparticle composition is between 2 and 7. In one embodiment, the apparent pKa of the lipid nanoparticle composition is 2, 3, 4, 5, 6, or 7. In one embodiment, the apparent pKa of the lipid nanoparticle composition is less than 6. In one embodiment, the apparent pKa of the lipid nanoparticle composition is between 3 and 6.

[1002] In one embodiment, the apparent pKa is determined by 2-(p-toluidinyl)-6-naphthalenesulfonic acid (TNS) fluorescence. TNS fluorescence has been described in the art, such as Jayaraman M, et al., Maximizing the potency of siRNA lipid nanoparticles for hepatic gene silencing in vivo. Angew Chem Int Ed, 2012. 51, 8529–8533, which is incorporated herein by reference.

[1003] 5.4 Pharmaceutical Compositions

[1004] In one embodiment, provided herein is a pharmaceutical composition comprising the lipid nanoparticles described in Section 0 or the lipid nanoparticle composition described in Section 0. In one embodiment, the pharmaceutical composition further comprises a pharmaceutically acceptable carrier.

[1005] Pharmaceutically acceptable carriers for the application include nontoxic carriers, adjuvants or supporting agents, which can not destroy the pharmacological activity of the compound prepared together. Pharmaceutically acceptable carriers that can be used for compositions of the present disclosure include, but are not limited to, ion exchangers, aluminum oxide, aluminum stearate, lecithin, serum proteins (for example, human serum albumin), buffer substances (such as phosphates), glycine, sorbic acid, potassium sorbate, mixtures of partial glycerides of saturated vegetable fatty acids, water, salt or electrolytes (such as protamine sulfate), disodium hydrogen phosphate, potassium hydrogen phosphate, sodium chloride, zinc salts, silica gel, magnesium trisilicate, polyvinyl pyrrolidone, cellulose-based materials, polyethylene glycol, sodium carboxymethyl cellulose, polyacrylates, waxes, polyethylene-polyoxypropylene block polymers, polyethylene glycol and lanolin.

[1006] In one embodiment, the pharmaceutical composition is formulated for oral administration. In one embodiment, the pharmaceutical composition is formulated for intravenous administration. In one embodiment, the pharmaceutical composition is formulated for intramuscular administration. In one embodiment, the pharmaceutical composition is formulated for inhalation administration. In one embodiment, administration is intra-arterial administration. In one embodiment, administration is intraperitoneal administration.

[1007] Typically, the pharmaceutical compositions provided herein are administered in an effective amount. The amount of the pharmaceutical composition actually administered will typically be determined by a physician based on relevant circumstances, including the condition to be treated or prevented, the selected route of administration, the actual pharmaceutical composition administered, the age, weight, and response of the individual patient, the severity of the patient's symptoms, and the like.

[1008] When used to prevent the conditions of the present disclosure, the pharmaceutical compositions provided herein will generally be administered to a subject at risk of developing the condition, at the dosage levels described above, on the advice and under the supervision of a physician. Subjects at risk of developing a particular condition generally include those with a family history of the condition, or those identified as being particularly susceptible to the condition through genetic testing or screening.

[1009] The pharmaceutical compositions provided herein can also be administered chronically ("chronic administration"). Chronic administration refers to administration of a compound or pharmaceutical composition thereof over an extended period of time (e.g., over a period of 3 months, 6 months, 1 year, 2 years, 3 years, 5 years, etc.), or can continue indefinitely (e.g., for the remainder of the subject's life). In certain embodiments, chronic administration is intended to provide a constant level of the compound in the blood, e.g., over an extended period of time, within a therapeutic window.

[1010] Pharmaceutical composition of the present disclosure can be further delivered using a variety of administration methods. For example, in certain embodiments, pharmaceutical composition can be given in the form of a single dose, for example, so that the concentration of compound in blood is increased to an effective level. The arrangement of a single dose depends on the systemic level of the active ingredient desired by the whole body, for example, intramuscular or subcutaneous single doses allow the slow release of active ingredient, and the single dose (for example, delivered by intravenous drip) delivered directly to the vein allows faster delivery, which rapidly increases the concentration of active ingredient in blood to an effective level. In other embodiments, pharmaceutical composition can be used in the form of continuous infusion, for example, by intravenous drip, to provide the maintenance of the steady-state concentration of active ingredient in the body of the experimenter. In addition, in other embodiments, pharmaceutical composition can first be used in the form of a single dose, then used in the form of continuous infusion.

[1011] Compositions for oral administration can take the form of bulk liquid solutions or suspensions or bulk powders. However, more commonly, compositions are presented in the form of unit dosage forms to facilitate accurate administration. The term "unit dosage form" refers to a physically discrete unit suitable for use as a single dose for human subjects and other mammals, each unit containing a predetermined amount of active material, which is calculated together with suitable pharmaceutical excipients to produce the desired therapeutic effect. Typical unit dosage forms include pre-filled, pre-measured ampoules or syringes of liquid compositions, or include pills, tablets, capsules, etc. in the case of solid compositions. In such compositions, the active substance is typically a minor component (approximately 0.1 to approximately 50% by weight, or alternatively approximately 1 to approximately 40% by weight), and the remainder is various carriers or excipients and processing aids that contribute to forming the desired form of administration.

[1012] Liquid forms suitable for oral administration may include a suitable aqueous or non-aqueous vehicle with buffering agents, suspending and dispensing agents, coloring agents, flavoring agents, etc. Solid forms may include, for example, any of the following ingredients or compounds of a similar nature: binders such as microcrystalline cellulose, tragacanth, or gelatin; excipients such as starch or lactose; disintegrants such as alginic acid, Primogel, or corn starch; lubricants such as magnesium stearate; glidants such as colloidal silicon dioxide; sweeteners such as sucrose or saccharin; or flavorings such as mint, methyl salicylate, or orange flavor.

[1013] Injectable compositions are generally based on injectable sterile saline or phosphate buffered saline or other injectable excipients known in the art. As before, the active compound in such compositions is usually a minor component, generally about 0.05 to 10% by weight, with the remainder being injectable excipients and the like.

[1014] The above components for orally administrable, injectable or topically administrable compositions are merely representative. Additional materials and processing techniques are described in Part 8 of Remington's Pharmaceutical Sciences, 17th edition, 1985, Mack Publishing Company, Easton, Pennsylvania, which is incorporated herein by reference.

[1015] Also provided herein is a kit (e.g., a pharmaceutical pack) comprising a pharmaceutical composition described herein. The kits described herein can include a pharmaceutical composition and other therapeutic or diagnostic or prophylactic agents, as well as a first and a second container (e.g., a vial, an ampoule, a bottle, a syringe, and / or dispersible packaging or other material) containing the pharmaceutical composition or other therapeutic or diagnostic or prophylactic agent. In some embodiments, the provided kits can also optionally include a third container containing a pharmaceutically acceptable excipient for diluting or suspending the nanoparticle composition of the present disclosure and / or other therapeutic or diagnostic or prophylactic agent. In some embodiments, the nanoparticle composition of the present application provided in the first container and the other therapeutic or diagnostic or prophylactic agent provided in the second container are combined to form a unit dosage form.

[1016] 5.5 Methods of Making or Producing Lipid Nanoparticles

[1017] The present invention also provides a method for producing lipid nanoparticles, comprising the following steps:

[1018] (i) dissolving a mixture comprising the lipids described herein in a first solution, wherein the lipid solution is formed in an organic solvent;

[1019] (ii) dissolving the payload in the second solution to form a payload solution; and

[1020] (iii) The lipid solution and the payload solution are mixed to form lipid nanoparticles.

[1021] In one embodiment, the organic solvent is a water miscible organic solvent. In one embodiment, the organic solvent is an alcohol. In one embodiment, the organic solvent is ethanol.

[1022] In one embodiment, the second solution is an aqueous solution. In one embodiment, the second solution is an aqueous solution having a pH below 7. In one embodiment, the second solution is an aqueous solution having a pH between 3 and 6. In one embodiment, the second solution is an aqueous solution having a pH of about 3.5, about 4, about 4.5, about 5, about 5.5, or about 6. In one embodiment, the second solution is a sodium acetate buffer solution having a pH of about 4.5.

[1023] 5.6 Treatment

[1024] In one embodiment, provided herein is a method of treating or preventing a disease or condition in a subject. In one embodiment, the method comprises administering to a subject a lipid nanoparticle as described in Section 0. In one embodiment, the method comprises administering to a subject a lipid nanoparticle composition as described in Section 0. In one embodiment, the method comprises administering to a subject a pharmaceutical composition as described in Section 0.

[1025] In one embodiment, provided herein is a method of treating or preventing a disease or condition in a subject, comprising administering to the subject a therapeutically effective amount of lipid nanoparticles comprising (i) a steroid compound (see Section 0) and (ii) an ionizable lipid (0).

[1026] In one embodiment, provided herein is a lipid nanoparticle for delivering or expressing a payload in the spleen of a subject, wherein the lipid nanoparticle is administered to the subject by a systemic administration route, wherein the lipid nanoparticle comprises an ionizable lipid and a steroid compound, and wherein the steroid compound comprises one or more cationic groups (e.g., the steroid compound in Section 0).

[1027] In one embodiment, administration is systemic administration. In one embodiment, administration is intravenous administration. In one embodiment, administration is intra-arterial administration. In one embodiment, administration is intraperitoneal administration. In one embodiment, administration is oral administration. In one embodiment, administration is intramuscular administration.

[1028] In one embodiment, provided herein is a method of treating a spleen disease in a subject. In one embodiment, provided herein is a method of treating a spleen-related disease in a subject.

[1029] In one embodiment, the spleen disease or spleen-related disease comprises a hematological disease, an infectious disease, an immune disease, or cancer.

[1030] In one embodiment, the spleen disease or spleen-related disease includes hemolytic anemia, thrombocytopenic purpura, chronic leukemia, lymphoma, such as Hodgkin's lymphoma or non-Hodgkin's lymphoma, myelodysplastic syndrome, myelofibrosis, splenomegaly, lipid metabolism disorders, such as glucocerebrosidosis and sphingomyelinosis, splenomegaly and hypersplenism associated with sepsis, typhoid fever, infectious mononucleosis or subacute bacterial endocarditis, splenomegaly and hypersplenism associated with malaria, tuberculosis, kala-azar, etc., congestive splenomegaly and hypersplenism caused by portal hypertension in cirrhosis, splenic cysts, such as parasitic splenic cysts or pseudosplenic cysts, splenic abscesses, primary splenic tumors, splenic metastases, splenic artery aneurysms, splenic infarction, splenic purpura, splenic rupture, etc.

[1031] In one embodiment, provided herein is a method of delivering or expressing a payload described in Section 0 in a subject, comprising administering a therapeutically effective amount of a lipid nanoparticle described in Section 0 or a lipid nanoparticle composition described in Section 0 to the subject.

[1032] In one embodiment, the amount of payload delivered or expressed in the spleen of a subject is higher than the amount of payload delivered or expressed in the liver of a subject. In one embodiment, the amount of payload delivered or expressed in the spleen of a subject is at least 2 times higher than the amount of payload delivered or expressed in the liver of a subject. In one embodiment, the amount of payload delivered or expressed in the spleen of a subject is at least 5 times higher than the amount of payload delivered or expressed in the liver of a subject. In one embodiment, the amount of payload delivered or expressed in the spleen of a subject is at least 10 times higher than the amount of payload delivered or expressed in the liver of a subject. In one embodiment, the amount of payload delivered or expressed in the spleen of a subject is at least 20 times higher than the amount of payload delivered or expressed in the liver of a subject. In one embodiment, the amount of payload delivered or expressed in the spleen of a subject is at least 40 times higher than the amount of payload delivered or expressed in the liver of a subject. In one embodiment, the amount of payload delivered or expressed in the spleen of a subject is at least 60 times higher than the amount of payload delivered or expressed in the liver of a subject. In one embodiment, the amount of payload delivered or expressed in the spleen of the subject is at least 100-fold greater than the amount of payload delivered or expressed in the liver of the subject.

[1033] In one embodiment, the amount of protein expressed in the spleen of a subject by the mRNA encapsulated in the LNP is higher than the amount of protein expressed in the liver of a subject by the mRNA encapsulated in the LNP. In one embodiment, the amount of protein expressed in the spleen of a subject by the mRNA encapsulated in the LNP is at least 5 times, at least 10 times, at least 20 times, at least 30 times, at least 40 times, at least 50 times, at least 60 times, at least 70 times, at least 80 times, at least 90 times, or at least 100 times higher than the amount of protein expressed in the liver of a subject by the mRNA encapsulated in the LNP.

[1034] In one embodiment, the ratio of the amount of payload delivered or expressed in the spleen of a subject to the amount of payload delivered or expressed in the liver of a subject (spleen / liver ratio) is greater than 1. In one embodiment, the spleen / liver ratio is greater than 2. In one embodiment, the spleen / liver ratio is greater than 5. In one embodiment, the spleen / liver ratio is greater than 10. In one embodiment, the spleen / liver ratio is greater than 15. In one embodiment, the spleen / liver ratio is greater than 20. In one embodiment, the spleen / liver ratio is greater than 25. In one embodiment, the spleen / liver ratio is greater than 30. In one embodiment, the spleen / liver ratio is greater than 35. In one embodiment, the spleen / liver ratio is greater than 40. In one embodiment, the spleen / liver ratio is greater than 50. In one embodiment, the spleen / liver ratio is greater than 60. In one embodiment, the spleen / liver ratio is greater than 70. In one embodiment, the spleen / liver ratio is greater than 80. In one embodiment, the spleen / liver ratio is greater than 90. In one embodiment, the spleen / liver ratio is greater than 100.

[1035] 6. Specific Implementation Methods

[1036] In order to make the technical solutions of the present disclosure clearer and more specific, the present disclosure is further described through the following examples. The following examples are only used to illustrate the specific embodiments of the present disclosure so that those skilled in the art can understand the present application, but are not intended to limit the scope of protection of the present application. Technical means or methods not specifically described in the specific embodiments of the present disclosure are conventional technical means or methods in the art. Unless otherwise specified, the materials, reagents, etc. used in the examples are commercially available.

[1037] Table 1 Abbreviations

[1038] Example 1.1: Synthesis of Compound 46

[1039] Potassium carbonate (1.55 g, 11.2 mmol, 4.0 eq.) was added to a solution of compound 1-6 (959 mg, 2.8 mmol, 1.0 eq.) and 3-1 (638 mg, 3.08 mmol, 1.1 eq.) in DMF. The mixture was heated at 60 ° C for 4 hours, then cooled to room temperature and quenched with a saturated aqueous sodium chloride solution. The resulting solution was extracted with ethyl acetate, and the organic phases were combined, dried over anhydrous sodium sulfate, filtered and concentrated to obtain a crude product, which was further purified by silica gel column chromatography to obtain compound 3-2 (682 mg).

[1040] Compound 3-2 (324mg, 0.69mmol, 1.0eq.) is dissolved in 5.0mL dichloromethane, and the reaction system is cooled to 0 DEG C in an ice bath. Add 2 drops of DMF, then oxalyl chloride (0.24mL, 2.8mmol, 4.0eq.) is added dropwise to the reaction solution. After dropwise addition is complete, the ice bath is removed, and the mixture is stirred at room temperature for 1 hour. Use rotary evaporator to remove the solvent, to give the crude acyl chloride product (309mg) in the form of oily matter, which is directly used in the next reaction step.

[1041] Compound 3-3 (407 mg, 1.9 mmol, 3.0 eq) was added to a solution of crude acid chloride (309 mg) in DCE (3.0 mL), and the reaction was heated to 70° C. to react overnight. The reaction solution was cooled to room temperature, and the solvent was removed using a rotary evaporator to give a crude product, which was purified by a silica gel column to give compound 3-4 (325 mg).

[1042] Compound 3-4 is dissolved in 4.0mL methanol, and at room temperature adds NaBH4(28mg, 0.73mmol).TLC indicates that starting material disappears.Mixture is quenched with saturated NaCl aqueous solution, extracted with dichloromethane (x3), dichloromethane is combined and passed through anhydrous sodium sulfate drying, filtered and concentrated under vacuum to obtain crude product 3-5 (260mg), which is used for next step without purification.

[1043] Crude compound 3-5 (260mg, 0.39mmol, 1.0eq.) is dissolved in 5.0mL dichloromethane, and compound 1-10 (77.4mg, 0.59mmol, 1.5eq.), EDCI (224mg, 1.17mmol, 3.0eq.), triethylamine (0.16mL, 1.17mmol, 3.0eq.) and DMAP (48mg, 0.39mmol, 1.0eq.) are added to reaction system. Reaction solution is stirred at room temperature for 12 hours. Then by adding saturated sodium chloride solution, reaction solution is quenched, and extracted with dichloromethane. Organic phase is combined, dried over anhydrous sodium sulfate, and filtered. Filtrate is collected and concentrated, to give crude product, which is passed through preparative high performance liquid chromatography to give compound 46 (32.6mg) in oily product form. 1 H NMR (400MHz, CDCl3): δppm 0.88(t,J=7.2Hz,9H),1.14(s,12H),1.15-1.28(m,37H),1.47-1.59(m,18H),1.75-1.84(m, 2H),2.24-2.35(m,10H),3.95(d,J=5.6Hz,2H),4.03(t,J=6.8Hz,2H),4.80-4.87(m,1H); MS m / z[M+H] + (ESI):780.7.

[1044] Example 1.2: Synthesis of Compound 132

[1045] To a 500 mL three-necked round-bottom flask at room temperature were added 1-nonanol (15 g, 104.0 mmol, 1.0 eq.), 8-bromooctanoic acid (25.5 g, 114.0 mmol, 1.1 eq.), DMAP (2.54 g, 20.8 mmol, 0.2 eq.), DIEA (40.3 g, 312.0 mmol, 3.0 eq.), and EDCI (25.9 g, 135 mmol, 1.3 eq.), followed by 250 mL of DCM. The reaction mixture was stirred at room temperature for 4 hours, and the reaction was monitored for completion by TLC. The reaction mixture was poured into 200 mL of saturated aqueous ammonium chloride and extracted with 3 x 100 mL of DCM. The organic phases were combined and dried over anhydrous sodium sulfate. The filtrate was filtered and concentrated to dryness to obtain the crude product, which was purified on a silica gel column to afford 21 g of compound 132-2 as a yellow oil.

[1046] Compound 132-2 (21 g, 60.1 mmol, 1.0 eq.) and 2-aminoethanol (110 g, 1.80 mol, 30.0 eq.) were added to a 250 mL three-necked round-bottom flask at room temperature and dissolved in 100 mL of methanol. The mixture was heated to 60°C and stirred for 18 hours. The solvent was removed by concentration, and saturated aqueous ammonium chloride and ethyl acetate were added. The organic phases were separated and extracted. The combined organic phases were dried over anhydrous sodium sulfate. The filtrate was filtered and concentrated to dryness to obtain the crude product, which was purified on a silica gel column to afford 14 g of compound 132-3 as a yellow oil.

[1047] To a 250 mL three-necked round-bottom flask at room temperature, 9-heptadecanol (9.8 g, 38.2 mmol, 1.0 eq.) and triethylamine (15.5 g, 152.8 mmol, 4.0 eq.) were added and dissolved in 100 mL of DCM. The reaction system was cooled in an ice bath, and then 2-methylpropanoyl chloride (9.8 g, 91.7 mmol, 2.4 eq.) was slowly added. The reaction was allowed to warm to room temperature overnight. TLC monitored the reaction for completion, and the reaction was quenched by adding saturated aqueous ammonium chloride. The mixture was extracted with DCM, and the organic phases were combined and dried over anhydrous sodium sulfate. The filtrate was filtered and concentrated to dryness to obtain the crude product, which was purified on a silica gel column to afford 8.7 g of compound 132-5 as a yellow oil.

[1048] To a 250 mL three-necked round-bottom flask at room temperature, compound 132-5 (8.7 g, 26.6 mmol, 1.0 eq.) was added and dissolved in 60 mL of THF. The reaction system was cooled to -40°C, and LDA (13.1 mL, 26.2 mmol, 0.98 eq.) was slowly added dropwise. The reaction was stirred for 1 hour. 1,6-Dibromohexane (9.03 g, 37.0 mmol, 1.39 eq.) and DMPU (0.48 g, 3.73 mmol, 0.14 eq.) were then added to the reaction system. The reaction was allowed to warm to room temperature overnight. TLC monitored the reaction completion, and the reaction was quenched by adding saturated ammonium chloride in ice water. The reaction was extracted with ethyl acetate, and the organic phases were combined and dried over anhydrous sodium sulfate. The filtrate was filtered and concentrated to dryness to obtain the crude product, which was purified on a silica gel column to afford 10.1 g of compound 132-6 as a yellow oil.

[1049] To a 20 mL round-bottom flask were added compound 132-6 (1.48 g, 3.04 mmol, 2.0 eq.), compound 132-3 (500 mg, 1.52 mmol, 1.0 eq.), K2CO3 (628.2 mg, 4.55 mmol, 3.0 eq.), KI (302.3 mg, 1.82 mmol, 1.2 eq.), cyclopentyl methyl ether (7.5 mL), and acetonitrile (2.5 mL). The temperature was raised to 80°C. Following completion of the reaction, TLC was used to quench the reaction. The reaction was then extracted with DCM, and the combined organic phases were dried over anhydrous sodium sulfate. The filtrate was filtered and concentrated to dryness to obtain the crude product, which was then purified on a silica gel column to afford 96.98 mg of compound 132 as an oil. 1 H NMR(400MHz, CDCl3)δ:4.86-4.80(m,1H),4.04(t,J=7.2Hz,2H),3.57(m,2H),2.62(t,J=7.2Hz,2H),2.49(t,J=7.2Hz,4H ),2.29(t,J=7.2Hz,2H),1.62(m,4H),1.51-1.45(m,10H),1.38-1.23(m,49H),1.15(s,6H),0.88(t,J=7.2Hz,9H); ESI-MS m / z:738.60[M+H] + .

[1050] Example 2: Synthesis of compound Lys-es-cholesterol

[1051] (pyridine: pyridine)

[1052] Cholesteryl chloroformate 2-1 (1.0 g, 2.2 mmol, 1.0 eq.) and pyridine (18 μL, 0.22 mmol, 0.1 eq.) were dissolved in 20 mL of dichloromethane. 1,6-hexanediol 2-2 (1.1 mL, 8.8 mmol, 4.0 eq.) was then added to the reaction mixture, which was then heated at 45 ° C for 12 hours. After the reaction, the reaction mixture was cooled to room temperature, quenched with a saturated sodium chloride solution, extracted with dichloromethane, and then combined and dried over anhydrous sodium sulfate. The solution was filtered and dried to obtain a crude product, which was purified by silica gel column chromatography to obtain intermediate compound 2-3 (879 mg).

[1053] Compound 2-4 (0.49g, 2.0mmol, 1.0eq.), pyridine (0.24mL, 3.0mmol, 1.5eq.) and DCC (0.62g, 3.0mmol, 1.5eq.) are dissolved in dichloromethane (20mL). Then, compound 2-3 (879mg, 1.7mmol, 0.85eq.) prepared above is added to reaction mixture. At room temperature, react until complete conversion is observed by TLC. Reaction mixture is quenched with saturated sodium chloride solution, extracted with dichloromethane, and organic phase is combined and dried with anhydrous sodium sulfate. The mixture is filtered, concentrated to obtain crude product, which is then purified by silica gel column chromatography to obtain intermediate compound 2-5 (641mg).

[1054] Compound 2-5 (200mg) is dissolved in the mixture of dichloromethane (4.0mL) and trifluoroacetic acid (2.0mL).Mixture is at room temperature stirred, until the complete reaction of compound 2-5 is realized.Then by rotary evaporation, organic solvent and trifluoroacetic acid are removed, thereby produce solid crude product, it is dispersed in the mixture of methyl alcohol and normal hexane.Then slurries are filtered and dried, to obtain final product Lys-es cholesterol (62.2mg). 1 H NMR (400MHz, CD3OD): δ0.74 (s, 3H), 0.88 (d, J = 6.8Hz, 6H), 0.90-1.68 (m, 38H), 1.75-2.18 (m, 8H), 2.28-2.38 (m, 2H), 2.96 MS m / z[M+H] + (ESI):659.5.

[1055] Example 3: Synthesis of Compound Arg-es-Cholesterol

[1056] The compound 2-5 (200mg, 0.26mmol, 1.0eq.) and compound 3-1 (113mg, 0.29mmol, 1.1eq.) prepared in Example 2 are dissolved in dichloromethane (5.0mL). Triethylamine (72 μ L, 0.52mmol, 2.0eq.) is added to the reaction solution, which is then reacted until the substrate is completely consumed by TLC analysis. The reaction solution is cooled to room temperature, and the reaction is quenched with a saturated sodium chloride solution, extracted with dichloromethane, and the organic phase is combined and dried over anhydrous sodium sulfate. The mixture is filtered and concentrated to obtain a crude product, which is further purified to obtain intermediate compound 3-2 (174mg) by silica gel column chromatography.

[1057] Compound 3-2 (174 mg) is dissolved in dichloromethane (4.0 mL) and formic acid (2.0 mL). The reaction mixture is stirred at room temperature until compound 3-2 most of disappears. Use rotary evaporator to remove solvent to obtain crude product. The crude product obtained is dispersed in a mixture of methanol and normal hexane, and then the slurries are filtered and dried to obtain the final product Arg-es-cholesterol (60.2 mg). 1 H NMR (400MHz, CD3OD): δ0.73(s,3H),0.88(d,J=6.8Hz,3H),0.94(d,J=6.8Hz,3H),1.05-1.44(m,14H),1.53-1.72(m,24H),1.74-2.08(m,8H),2 MS m / z[M+H] + (ESI):701.5.

[1058] Example 4: Synthesis of Compound MHEM-Cholesterol

[1059] (RT: room temperature; toluene: toluene; reflux: reflux; MHEM-Cholesterol: MHEM-cholesterol)

[1060] Compound 2-1 (450mg, 1.0mmol, 1.0eq.) and 2-bromoethylamine hydrobromide (compound 4-2, 225mg, 1.1mmol, 1.1eq.) are dissolved in 10mL chloroform. The reaction mixture is cooled to -30 DEG C, then triethylamine (0.21mL, 1.5mmol, 1.5eq.) is added to the reaction solution. After 30 minutes, the temperature is raised to room temperature, and under agitation, the reaction is continued until compound 2-1 is completely consumed. The reaction solution is quenched with saturated sodium chloride solution, and extracted with dichloromethane. The organic phase is combined, dried over anhydrous sodium sulfate, then organic solvent is removed by rotary evaporation to obtain thick intermediate compound 4-3 (452mg), which is directly used in the next step without purification.

[1061] The compound 4-3 (452 ​​mg, 0.84 mmol, 1.0 eq.) and compound 4-4 (89 mg, 1.0 mmol, 1.2 eq.) p...

Claims

1. A lipid nanoparticle comprising an ionizable lipid and a steroid compound, wherein the steroid compound is a compound of formula (I): or a stereoisomer, a mixture of stereoisomers, or a pharmaceutically acceptable salt thereof, wherein: Is a single bond or a double bond; Y is -C(O)O-, -O-, -NH-, -SC(O)O-, -OC(O)NR 2a -、-NR 2a C(O)NR 2a -、-OC(O)S-、-OC(O)O-、-NR 2a C(O)O-, -OC(O)-, -SC(O)-, -C(O)S-, -NR 2a -、-C(O)NR 2a -、-NR 2a C(O)-、-NR 2a C(O)S-、-SC(O)NR 2a -, -C(O)-, -OC(S)-, -C(S)O-, -OC(S)NR 2a -、-NR 2a C(S)O- or -S(O) 0-2 -, and wherein the leftward connection is in the direction of V1; Y1 is absent, -C(O)O-, -O-, -NH-, -SC(O)O-, -OC(O)NR 2a -、-NR 2a C(O)NR 2a -、-OC(O)S-、-OC(O)O-、-NR 2a C(O)O-, -OC(O)-, -SC(O)-, -C(O)S-, -NR 2a -、-C(O)NR 2a -、-NR 2a C(O)-、-NR 2a C(O)S-、-SC(O)NR 2a -, -C(O)-, -OC(S)-, -C(S)O-, -OC(S)NR 2a -、-NR 2a C(S)O- or -S(O) 0-2 -, and wherein the leftward connection is in the direction of V1; R 2a Each instance of is independently H, C 1-10 Alkyl, C 3-14 cycloalkyl or 3 to 14 membered heterocyclyl, and wherein the alkyl, cycloalkyl and heterocyclyl are independently optionally replaced by C 1-10 Alkyl, L 2b -OR 2b , L b -SR 2b , L 2b -NR 2b R' 2b , L 2b N + R 2b R' 2b R” 2b , L 2b -OC(O)R 2b or L 2b -C(O)OR 2b replace; R 201 、R 202、 R 203 、R 204 Each example is independently H, OH, alkyl, alkoxy, -L 2b -NR 21 R 22 、-L 2b -N + R 21 R 22 R 23 , L 2b -OC(O)R 21 or L 2b -C(O)OR 21 ; m is an integer from 1 to 12; m1 is an integer from 0 to 12; V1 is NR 21 R 22 、N + R 21 R 22 R 23 , imidazole, pyridine, pyrrole, pyrrolidine, pyrazole, (iso)thiazole, (iso)oxazole, oxadiazine, oxazoline, dithioureaazole, (iso)triazole, tetrazole, pentazole, indole, indoline, pyrimidine, pyrazine, quinazoline, piperazine, piperidine, morpholine, pyran, quinoxaline, quinoline, quinolinone, (iso)quinoline, amidine, guanidine or urea; M is absent or is one or more anions independently selected from chloride, bromide, iodide, sulfate, nitrate, perchlorate, formate, citrate, sulfonate, methanesulfonate, trifluoromethanesulfonate, trifluoroacetate, pyrosulfate, bisulfate, sulfite, bisulfite, phosphate, pyrophosphate, monohydrogen phosphate, dihydrogen phosphate, acetate, propionate, decanoate, octanoate, acrylate, isobutyrate, hexanoate, heptanoate, oxalate, malonate, succinate, fumarate, maleate, benzoate, phthalate, phenylacetate, lactate, glycolate, tartrate, mandelate, methanesulfonate, gluconate, aspartate, or glutamate; R 21 、R 22 、R 23 Each instance of is independently H, optionally substituted C 1-8 Alkyl, -L 2b -OR 2b 、-L 2b -SR 2b 、-L 2b -NR 2b R' 2b R” 2b , L 2b -OC(O)R 2b or L 2b -C(O)OR 2b ; L 2b is absent or optionally substituted C 1-10 alkylene; R 2b , R' 2b and R” 2b Each instance of is independently H or C 1-10 alkyl; R z1 It is C 1-14 Alkyl or C 1-14 alkenyl, wherein the alkyl and alkenyl groups are optionally substituted; and Provided that when Y is -NHC(O)-, at least one of the following conditions is met: (i) m is an integer from 5 to 12; or (ii) V1 is not N(CH3)2.

2. A lipid nanoparticle comprising an ionizable lipid and a steroid compound, wherein the steroid compound is a compound of formula (IP): or a stereoisomer, a mixture of stereoisomers, or a pharmaceutically acceptable salt thereof, wherein: Is a single bond or a double bond; Y1 does not exist, -C(O)O-, -O-, -NH-, -SC(O)O-, -OC(O)NR 2a -、-NR 2a C(O)NR 2a -、-OC(O)S-、-OC(O)O-、-NR 2a C(O)O-, -OC(O)-, -SC(O)-, -C(O)S-,-NR 2a -,-C(O)NR 2a -,-NR 2a C(O)-,-NR 2a C(O)S-,-SC(O)NR 2a -,-C(O)-,-OC(S)-, -C(S)O-, -OC(S)NR 2a -、-NR 2a C(S)O-, -S(O) 0-2 -or -OPO3-, and wherein the leftward connection is in the direction of V1; R 2a Each instance of is independently H, C 1-10 Alkyl, C 3-14 cycloalkyl or 3 to 14 membered heterocyclyl, and wherein the alkyl, cycloalkyl and heterocyclyl are independently optionally replaced by C 1-10 Alkyl, L 2b -OR 2b , L b -SR 2b , L 2b -NR 2b R' 2b , L 2b N + R 2b R' 2b R” 2b , L 2b -OC(O)R 2b or L 2b -C(O)OR 2b replace; R 201 、R 202、 R 203 、R 204 Each example is independently H, OH, alkyl, alkoxy, -L 2b -NR 21 R 22 、 -L 2b -N + R 21 R 22 R 23 、L 2b -OC(O)R 21 或L 2b -C(O)OR 21 ; R 205 Absence, cations, H, C 1-6 Alkyl, C 1-6 Halogenated alkyl, C 2-6 Alkenyl or C 2-6 alkynyl, and wherein alkyl, haloalkyl, alkenyl and alkynyl are optionally substituted; m is an integer from 1 to 12; m1 is an integer from 0 to 12; V1 is NR 21 R 22 、N + R 21 R 22 R 23 , imidazole, pyridine, pyrrole, pyrrolidine, pyrazole, (iso)thiazole, (iso)oxazole, oxadiazine, oxazoline, dithioureaazole, (iso)triazole, tetrazole, pentazole, indole, indoline, pyrimidine, pyrazine, quinazoline, piperazine, piperidine, morpholine, pyran, quinoxaline, quinoline, quinolinone, (iso)quinoline, amidine, guanidine or urea; M is absent or is one or more anions independently selected from chloride, bromide, iodide, sulfate, nitrate, perchlorate, formate, citrate, sulfonate, methanesulfonate, trifluoromethanesulfonate, trifluoroacetate, pyrosulfate, bisulfate, sulfite, bisulfite, phosphate, pyrophosphate, monohydrogen phosphate, dihydrogen phosphate, acetate, propionate, decanoate, octanoate, acrylate, isobutyrate, hexanoate, heptanoate, oxalate, malonate, succinate, fumarate, maleate, benzoate, phthalate, phenylacetate, lactate, glycolate, tartrate, mandelate, methanesulfonate, gluconate, aspartate, or glutamate; R 21 、R 22 、R 23 Each instance of is independently H, optionally substituted C 1-8 Alkyl, -L 2b -OR 2b 、-L 2b -SR 2b 、-L 2b -NR 2b R' 2b R” 2b , L 2b -OC(O)R 2b or L 2b -C(O)OR 2b ; L 2b is absent or optionally substituted C 1-10 alkylene; R 2b , R' 2b and R” 2b Each instance of is independently H or C 1-10 alkyl; R z1 It is C 1-14 Alkyl or C 1-14 alkenyl, wherein the alkyl and alkenyl groups are optionally substituted.

3. The lipid nanoparticle of claim 1, wherein Y is -C(O)O-, -OC(O)-, -O-, -NHC(O)O-, -OC(O)NH- or -OC(O)O-, and wherein the leftward connection is in the direction of V1.

4. The lipid nanoparticle of any one of claims 1 to 3, wherein Y1 is absent, -C(O)O-, -OC(O)-, -O-, -S-, SO, S(O)2, -O(CO)O-, -NHC(O)- or -C(O)NH-, and wherein the leftward connection is in the direction of V1.

5. The lipid nanoparticle of claim 1, wherein the steroid compound is a compound of formula (I-A1), (I-A2), (I-A3), (I-A4) or (I-A5): or a stereoisomer, a mixture of stereoisomers, or a pharmaceutically acceptable salt thereof.

6. The lipid nanoparticle of claim 5, wherein the steroid compound is a compound of formula (I-B1), (I-B2), (I-B3), (I-B4), (I-B5), (I-B6), (I-B7) or (I-B8): or a stereoisomer, a mixture of stereoisomers, or a pharmaceutically acceptable salt thereof, wherein m3 is an integer from 1 to 11.

7. The lipid nanoparticle according to any one of claims 1 to 6, wherein R 201 、R 202 、R 203 and R 204 Each instance of is independently H, C 1-6 Alkyl, -NR 21 R 22 、-N + R 21 R 22 R 23 、-(CH2) 0-6 -OC(O)R 21 or -(CH2) 0-6 -C(O)OR 21 .

8. The lipid nanoparticle according to claim 7, wherein R 201 、R 202 、R 203 and R 204 Each instance of is independently H, NH2, NH3 + , -CH2OC(O)CH3, -CH2C(O)OCH3, -OC(O)CH3 or -C(O)OCH3.

9. The lipid nanoparticle according to any one of claims 1 to 8, wherein m is an integer from 5 to 12.

10. The lipid nanoparticle of claim 2, wherein the steroid compound is a compound of formula (I-P1): or a stereoisomer, a mixture of stereoisomers, or a pharmaceutically acceptable salt thereof, and wherein m3 is an integer from 1 to 11.

11. The lipid nanoparticle according to any one of claims 1 to 10, wherein V1 is NR 21 R 22 、N + R 21 R 22 R 23 , or an optionally substituted radical of amidine, guanidine or urea.

12. The lipid nanoparticle according to any one of claims 1 to 11, wherein R 21 、R 22 、R 23 are independently H, hydroxyl, C 1-3 Alkyl or C 1-3 Hydroxyalkyl.

13. The lipid nanoparticle according to any one of claims 1 to 12, wherein V1 is NH2, NH3 + 、N(CH3)2、N + (CH3)3、N + CH3(CH2CH2OH)2、N + (CH3)2(CH2CH2OH), NHC(=N + H2)NH2, NHC(O)NH2, CH2C(=NH)NH2, N(CH3)(CH2CH2CO2CH3), N + (CH3)2(CH2CH2CO2CH3), N(CH3)(CH2CO2CH3) or N + (CH3)2(CH2CO2CH3).

14. The lipid nanoparticle of claim 13, wherein V1 is N + CH3(CH2CH2OH)2、N + (CH3)2(CH2CH2OH) or N + (CH3)3.

15. The lipid nanoparticle of any one of claims 1 to 14, wherein M is absent, or one or more of bromide, chloride, iodide or trifluoroacetate.

16. The lipid nanoparticle according to any one of claims 1 to 5, wherein R z1 It is C 1-14 Alkyl or C 1-14 Alkenyl, wherein the alkyl and alkenyl groups are optionally substituted with one or more hydroxy, oxo, nitro, cyano, halogen, C 1-6 Alkoxy, C2-C6 alkenyl, C2-C6 alkynyl, C3-C8 cycloalkyl, C6-C 10 aryl, 5- to 10-membered heteroaryl, or 3- to 8-membered heterocyclic group.

17. The lipid nanoparticle according to claim 14, wherein R z1 It is C 6-12 Alkyl or C 6-12 and wherein the alkyl and alkenyl groups are optionally substituted with one or more hydroxy, oxo or halogen.

18. The lipid nanoparticle according to claim 17, wherein R z1 yes 19. A lipid nanoparticle comprising an ionizable lipid and a steroid compound, wherein the steroid compound is: or a stereoisomer, a mixture of stereoisomers, or a pharmaceutically acceptable salt thereof.

20. The lipid nanoparticle according to any one of claims 1 to 19, wherein the molar percentage of the steroid compound based on the total lipids is 3-15 mol%, preferably 5-10 mol%, such as 3 mol%, 4 mol%, 5 mol%, 6 mol%, 7 mol%, 8 mol%, 9 mol%, 10 mol%, 11 mol%, 12 mol%, 13 mol%, 14 mol% or 15 mol%.

21. The lipid nanoparticle of any one of claims 1 to 20, wherein the ionizable lipid comprises at least one ionizable head group and at least one biodegradable hydrophobic chain, wherein: The ionizable lipid has a logP of at least about 10.1 and a pKa of about 4 to about 11; and The structural formula of the biodegradable hydrophobic chain is -R 16 -M0-R 17 ;in R 16 It is C 4-14 Alkylene, C 4-14 Alkenylene or C 4-14 Alkynylidene; R 17 It is C 6-20 alkyl; and M0 is a biodegradable group, and wherein the alkylene, alkenylene, alkynylene and alkyl are optionally substituted; Preferably, M0 is an ester group, an amide group, a thioester group, a carbonate group, a carbamate group, a carbamoylthioester group, a urea group or a disulfide group.

22. The lipid nanoparticle of any one of claims 1 to 21, wherein the ionizable lipid is a compound of formula (III): or a stereoisomer, mixture of stereoisomers, tautomer, isotopologue or pharmaceutically acceptable salt thereof, wherein: Each instance of R3 and R4 is independently C 1-10 Alkyl, C 2-10 Alkenyl, C 2-10 Alkynyl, C3-C 14 Cycloalkyl, 3 to 14 membered heterocyclic group, (C 1-10 Alkylene)-(C3-C 14 Cycloalkyl) or (C 1-10 alkylene)-(3 to 14 membered heterocyclyl), or R3 and R4 together with the nitrogen atom to which they are attached form a 3 to 14 membered heterocyclyl, or R4 and one R0' together with the atom to which they are attached form a 4 to 10 membered heterocyclyl or a 5 to 10 membered heteroaryl, and wherein said alkyl, alkenyl, alkynyl, cycloalkyl, heterocyclyl, heteroaryl and alkylene are independently optionally replaced by one or more R * replace; Each instance of R0' is independently optionally substituted methylene, or wherein two R0' together with the carbon to which they are attached form a 3 to 8 membered cycloalkylene, and wherein the cycloalkylene is optionally substituted by one or more R ** replace; k is 0, 1, 2, 3, 4, 5, or 6; j is 0 or 1; The dotted line connecting Q and W is either non-existent or a bond; W is C, CH or N, provided that when W is N, j is 0 and the dotted bond connecting Q and W is absent; G5 does not exist, C 1-24 Alkylene, C 2-24 C3-C8 cycloalkylene, C3-C8 cycloalkenylene, wherein the alkylene, alkenylene, cycloalkylene, cycloalkenylene are optionally substituted by one or more R ** replace; G1, G2, G3 and G4 are each independently absent, C 1-13 Alkylene, C 2-13 Alkenylene or C 2-13 Alkyne, wherein the alkylene, alkenylene and alkynylene groups are optionally substituted by one or more R s replace; The total length of G1 and G2 is 3, 4, 5, 6, 7, 8, 9, 10, 11, 12 or 13 carbon atoms; The total length of G3 and G4 is 3, 4, 5, 6, 7, 8, 9, 10, 11, 12 or 13 carbon atoms; R5, R6, R7 and R8 are each independently hydrogen or optionally substituted C 1-8 alkyl; When the dotted line connecting Q and W is absent, Q is absent or is -C(O)O-, -O-, -NH-, -SC(O)O-, -OC(O)NR b -、-NR b C(O)NR b -、-OC(O)S-、-OC(O)O-、-NR b C(O)O-, -OC(O)-, -SC(O)-, -C(O)S-, -NR b -、-C(O)NR b -、-NR b C(O)-、-NR b C(O)S-、-SC(O)NR b -, -C(O)-, -OC(S)-, -C(S)O-, -OC(S)NR b -、-NR b C(S)O-, -SS-, or -S(O) 0-2 -, where the rightward connection is in the direction of W; When the dashed line connecting Q and W is a bond, G5 is absent, and Q and W form a 5- to 10-membered monocyclic or fused ring, optionally wherein the ring is separated by one or more R ** replace; M1 and M2 are each independently absent, or -C(O)O-, -O-, -SC(O)O-, -OC(O)NR a -、-NR a C(O)NR a -、-OC(O)S-、-OC(O)O-、-NR a C(O)O-, -OC(O)-, -SC(O)-, -C(O)S-, -NR a -、-C(O)NR a -、-NR a C(O)-、-NR a C(O)S-、-SC(O)NR a -, -C(O)-, -OC(S)-, -C(S)O-, -OC(S)NR a -、-NR a C(S)O-, -SS-, or -S(O) 0-2 -; Each instance of R0 is independently -(CRR')-; Each instance of R and R' is independently H, C 1-20 Alkyl, L a -OR a 、-L a -SR a or -L a -NR a R' a , or R and R' together with the carbon to which they are attached form C 3-8 cycloalkylene; Q3 and Q4 are each independently H, -(CRR')-, C 6-10 an aryl or steroid moiety; Each instance of A1, A2, A3, A4 is independently -(CR 18 R 18 -CR 18 =CR 18 )-or-(CR 18 R 18 -C≡C)-; R 18 Each instance of is independently H or C 1-20 alkyl; Each instance of N1 and N2 is independently a biodegradable group; Z does not exist, C 1-10 Alkylene or -OP(O)(OH)-O-; Each of the dashed lines between Z-Q3 or Z-Q4 is independently absent or a bond, provided that when Z is absent, both dashed lines are absent; m, n, q, r, u, v, y and z are each independently 0, 1, 2, 3, 4, 5, 6, 7, 8, 9 or 10; o, p, w and x are each independently 0, 1 or 2; s and t are each independently 0 or 1; The number of carbon atoms between G1 and Q3 or between G3 and Q4 is 8 to 30; L a and L e Each independently does not exist or is C 1-20 alkylene; L b and L f Each independently does not exist or is C 1-10 alkylene; L c Does not exist or C 1-8 alkylene; L d Does not exist or C 1-14 alkylene; R * is hydroxyl, halogen, C 1-10 Alkyl, C 1-10 Halogenated alkyl, C 1-10 Hydroxyalkyl, -L b -OR b 、-L b -SR b or -L b -NR b R' b ; R ** is hydroxyl, halogen, C 1-8 Alkyl, C 1-8 Halogenated alkyl, C 1-8 Hydroxyalkyl, -L b -OR b 、-L b -SR b or -L b -NR b R' b ; R s It is C 1-14 Alkyl, -L b -OR b 、-L b -SR b or -L b -NR b R' b ; R a and R' a Each independently is H, C 1-20 Alkyl, C 3-14 Cycloalkyl or 3 to 14 membered heterocyclyl, wherein the alkyl, cycloalkyl and heterocyclyl are optionally substituted by one or more selected from hydroxy, halogen, C 1-20 Alkyl, -L e -OR e 、-L e -SR e or -L e -NR e R' e The group substitution; R b and R' b Each independently is H, C 1-10 Alkyl, C 3-14 Cycloalkyl or 3 to 14 membered heterocyclyl, wherein the alkyl, cycloalkyl and heterocyclyl are optionally substituted by one or more selected from hydroxy, halogen, C 1-10 Alkyl, -L f -OR f 、-L f -SR f or -L f -NR f R' f The group substitution; R c and R' c are each independently hydrogen or C 1-8 alkyl; R d and R' d are each independently hydrogen or C 1-14 alkyl; R e and R' e are each independently hydrogen or C 1-20 alkyl; and R f and R' f are each independently hydrogen or C 1-10 alkyl.

23. The lipid nanoparticle of claim 22, wherein the ionizable lipid is a compound of formula (IV'): or a stereoisomer, a mixture of stereoisomers or a pharmaceutically acceptable salt thereof, wherein j is 0 or 1; W is CH or N, provided that when W is N, j is 0; k is 0, 1, 2, 3, 4, 5, 6, 7, or 8; Each instance of R0' is independently optionally substituted methylene, or wherein two R0' together with the carbon to which they are attached form a 3 to 8 membered cycloalkylene, and wherein the cycloalkylene is optionally substituted by one or more R ** replace; M1 and M2 are each independently -C(O)O-, -O-, -SC(O)O-, -OC(O)NR a -、-NR a C(O)NR a -、-OC(O)S-、-OC(O)O-、-NR a C(O)O-, -OC(O)-, -SC(O)-, -C(O)S-, -NR a -、-C(O)NR a -、-NR a C(O)-、-NR a C(O)S-、-SC(O)NR a -, -C(O)-, -OC(S)-, -C(S)O-, -OC(S)NR a -、-NR a C(S)O-, -SS-, or -S(O) 0-2 -; Q is absent, -C(O)O-, -O-, -SC(O)O-, -OC(O)NR b -, -NR b C(O)NR b -, -OC(O)S-, -OC(O)O-, -NR b C(O)O-, -OC(O)-, -SC(O)-, -C(O)S-, -NR b -, -C(O)NR b -, -NR b C(O)-, -NR b C(O)S-, -SC(O)NR b -, -C(O)-, -OC(S)-, -C(S)O-, -OC(S)NR b -, -NR b C(S)O-, -S-S- or -S(O) 0-2 -; R a and R b Each independently is H, C 1-20 Alkyl, C 3-14 Cycloalkyl or 3 to 14 membered heterocyclyl, wherein the alkyl, cycloalkyl and heterocyclyl are optionally substituted by one or more selected from hydroxy, halogen, C 1-20 Alkyl, -L e -OR e 、-L e -SR e or -L e -NR e R' e The group substitution; G5 is absent or optionally substituted C 1-8 alkylene; R1 and R2 are each independently C 4-20 Alkyl, C 4-20 Alkenyl or C 4-20 Alkynyl, wherein one or more methylene units in R1 and R2 are independently optionally replaced by -NH- or -N(C 1-20 alkyl), wherein the alkyl, alkenyl and alkynyl groups are optionally substituted; Each instance of R3 and R4 is independently H, C 1-10 Alkyl, C 2-10 Alkenyl, C 2-10 Alkynyl, C3-C 14 cycloalkyl or 3 to 14 membered heterocyclyl, or R3 and R4 together with the N atom to which they are attached form a 3 to 14 membered heterocyclyl, or R4 and one R0' together with the atom to which they are attached form a 4 to 10 membered heterocyclyl; and wherein said alkyl, alkenyl, alkynyl, cycloalkyl, heterocyclyl is optionally replaced by one or more R * replace; and R5, R6, R7 and R8 are each independently hydrogen or optionally substituted C 1-8 alkyl.

24. The lipid nanoparticle of claim 23, wherein the ionizable lipid is a compound of formula (V'), (VI') or (VII'): or a stereoisomer, a mixture of stereoisomers or a pharmaceutically acceptable salt thereof, wherein k is 0, 1, 2, 3, 4, or 5; L1 and L2 are each independently -(CRR')2-, -CH=CH-, -C≡C- or -NR"-; One of L3 and L5 is -(CR s R s ')2-, -CH=CH- or -C≡C-, and the other is absent; One of L4 and L6 is -(CR s R s ')2-, -CH=CH- or -C≡C-, and the other is absent; G 1a , G 1b , G 2a , G 2b , G 3a , G 3b , G 4a and G 4b Each of which is independently absent or optionally substituted C 1-7 alkylene; G 1a , G 1b , G 2a and G 2b The total length of is 1, 2, 3, 4, 5, 6 or 7 carbon atoms; G 3a , G 3b , G 4a and G 4b The total length of is 1, 2, 3, 4, 5, 6 or 7 carbon atoms; R5, R6, R7 and R8 are each independently hydrogen or optionally substituted C 1-6 alkyl; G7, G8, G9 and G 10 Each independently absent or optionally substituted C 1-12 Alkylene, provided that the total length of G7 and G8 is 4, 5, 6, 7, 8, 9, 10, 11 or 12 carbon atoms, and G9 and G 10 has a total length of 4, 5, 6, 7, 8, 9, 10, 11 or 12 carbon atoms; R s and R s 'Each independently is H, C 1-10 Alkyl, -L d -OR d or -L d -NR d R' d ; R' is H, C 1-14 Alkyl, -L a -OR a or -L a -NR a R' a ; R" is H or C 1-14 alkyl; L d Does not exist or C 1-10 alkylene; L a Does not exist or C 1-14 alkylene; R a and R' a Each independently is H, C 1-14 Alkyl, C 3-10 cycloalkyl or 3 to 10 membered heterocyclyl, and wherein the alkyl, cycloalkyl and heterocyclyl are optionally substituted; R d and R' d are each independently H or C 1-10 alkyl; a' and b are each independently 0, 1, 3, 4, or 5, provided that at least one of a' and b is not 0; g is 0, 1, 2, 3, 4, or 5; a'+g is 0, 1, 2, 3, 4 or 5; and c, d, e and f are each independently 0, 1, 2, 3, 4, 5, 6 or 7, provided that c+d is an integral between 2 and 9, and e+f is an integral between 2 and 9.

25. The lipid nanoparticle of any one of claims 1-24, wherein the ionizable lipid is a compound of formula (VIII): or a stereoisomer, a mixture of stereoisomers, or a pharmaceutically acceptable salt thereof.

26. The lipid nanoparticle of claim 25, wherein R5, R6, R7 and R8 are each independently hydrogen or methyl.

27. The lipid nanoparticle of claim 25, wherein G1, G2, G3, and G4 are each independently absent or C 1-8 Alkylene.

28. The lipid nanoparticle of any one of claims 1-27, wherein the ionizable lipid is a compound of formula (IX): or a stereoisomer, a mixture of stereoisomers or a pharmaceutically acceptable salt thereof, wherein G1 and G2 are each independently a bond, C 1-13 Straight chain alkylene, C 2-13 Straight chain alkenylene or C 2-13 Straight chain alkynylene, wherein the alkylene, alkenylene and alkynylene groups are optionally substituted by one or more R G1 replace; The total length of G1 and G2 is 3, 4, 5, 6, 7, 8, 9, 10, 11, 12 or 13 carbon atoms; R G1 In each case, independently H, C 1-14 Alkyl, -L a -OR a 、-L a -SR a or -L a -NR a R' a ; G3 is C 4-14 Straight chain alkylene, C 4-14 Straight chain alkenylene or C 4-14 Straight chain alkynylene, wherein the alkylene, alkenylene and alkynylene groups are optionally substituted by one or more R G3 replace; R G3 In each case, independently H, -L a -OR a 、-L a -SR a or -L a -NR a R' a ; L a independently a bond or C 1-14 alkylene; R a and R' a Each independently is H, C 1-14 Alkyl, C 3-14 Cycloalkyl or 3-14 membered heterocyclic group; G4 is the key, C 1-6 Alkylene, C 2-6 Alkenylene or C 2-6 Alkyne, wherein the alkylene, alkenylene and alkynylene groups are optionally substituted by one or more R G4 replace; R G4 In each case, independently H, C 1-6 Alkyl, -L b -OR b 、-L b -SR b or -L b -NR b R' b ; L b independently a bond or C 1-6 alkylene; R b and R' b Independently H, C 1-6 Alkyl, C 3-10 Cycloalkyl or 3-10 membered heterocyclic group; Or two R G4 Together with the carbon atom to which they are attached, they form C 3-14 Cycloalkylene or 3-14 membered heterocyclylene, wherein the cycloalkylene and heterocyclylene are optionally substituted by one or more R 4g replace; R 4g is independently H, halogen, cyano, C 1-8 Alkyl, C 1-8 Halogenated alkyl, -L e -OR e 、-L e -SR e or -L e -NR e R' e ; L e independently a bond or C 1-8 alkylene; R e and R' e Independently H, C 1-8 Alkyl, C 3-14 Cycloalkyl or 3-14 membered heterocyclic group; M1 and M2 are independently -C(O)O-, -OC(O)-, -O-, -SC(O)O-, -OC(O)NR-, -NRC(O)NR-, -OC(O)S-, -OC(O)O-, -NRC(O)O-, -SC(O)-, -C(O)S-, -NR-, -C(O)NR-, -NRC(O)-, -NRC(O)S-, -SC(O)NR-, -C(O)-, -OC(S)-, -C(S)O-, -OC(S)NR-, -NRC(S)O-, -S-S- or -S(O) 0-2 -; Q is a bond, -C(O)O-, -O-, -SC(O)O-, -OC(O)NR f -、-NR f C(O)NR f -、-OC(O)S-、-OC(O)O-、-NR f C(O)O-, -OC(O)-, -SC(O)-, -C(O)S-, -NR f -、-C(O)NR f -、-NR f C(O)-、-NR f C(O)S-、-SC(O)NR f -, -C(O)-, -OC(S)-, -C(S)O-, -OC(S)NR f -、-NR f C(S)O-, -SS-, -S(O) 0-2 -, phenylene or pyridinylene, wherein the phenylene or pyridinylene is optionally substituted with one or more R*; R* is independently H, halogen, cyano, C 1-10 Alkyl, C 1-10 Halogenated alkyl, -L f -OR f 、-L f -SR f or -L f -NR f R' f ; L f independently a bond or C 1-8 alkylene; R f and R' f Independently H, C 1-10 Alkyl, C 3-14 Cycloalkyl or 3-14 membered heterocyclic group; R1 and R2 are independently C 4-20 Alkyl, C 4-20 Alkenyl or C 4-20 Alkynyl, which is optionally substituted by one or more R 1s substituted, and wherein one or more methylene units are optionally and independently replaced by -NR'-; R 1s Independently H, C 1-20 Alkyl, -L c -OR c 、-L c -SR c or -L c -NR c R' c ; R and R' are independently H or C 1-20 alkyl; L c independently a bond or C 1-20 alkylene; R c and R' c Independently H, C 1-20 Alkyl, C 3-14 Cycloalkyl or 3-14 membered heterocyclic group; R3 is -CN, -OR g 、-C(O)R g 、-OC(O)R g 、-NR”C(O)R g 、-NR g R’ g 、-NR”C(O)NR g R’ g 、-NR”C(O)R g 、-NR”S(O)2R g 、-OC(O)NR g R’ g 、-NR”C(O)OR g 、-N(OR g )C(O)R g 、-N(OR g )S(O)2R g 、-N(OR g )C(O)OR g 、-N(OR g )C(O)R g R’ g 、a 3- to 14-membered heterocyclic group or a 5- to 14-membered heteroaryl group; R g and R' g Independently H, C 1-10 Alkyl, C 3-10 Cycloalkyl and 3-10 membered heterocyclic groups; R" is independently H or C 1-6 alkyl; R5 and R6 are independently C 1-8 Alkyl, wherein the alkyl group is optionally substituted by one or more R 4s replace; Or R5 and R6 together with the carbon atom to which they are attached form C 3-14 Cycloalkylene or 3-14 membered heterocyclylene, wherein the cycloalkylene and heterocyclylene are optionally substituted by one or more R 4s replace; R 4s are independently H, halogen, cyano, C 1-8 Alkyl, C 1-8 Halogenated alkyl, -L d -OR d 、-L d -SR d or -L d -NR d R' d ; L d independently a bond or C 1-8 alkylene; R d and R' d Independently H, C 1-8 Alkyl, C 3-14 Cycloalkyl or 3-14 membered heterocyclic group.

29. The lipid nanoparticle of any one of claims 1-28, wherein the ionizable lipid is a compound of formula (X): or a stereoisomer, a mixture of stereoisomers or a pharmaceutically acceptable salt thereof, wherein a is 1, 2, 3, 4, 5, or 6; b is 4, 5, 6, 7, 8, 9, or 10; c is 1, 2, 3, 4, 5, or 6; d is 0, 1, 2, 3, or 4; c+d is 3, 4, 5, 6, 7, 8 or 9.

30. The lipid nanoparticle of any one of claims 1 to 29, wherein the ionizable lipid is: or a stereoisomer, a mixture of stereoisomers, or a pharmaceutically acceptable salt thereof.

31. The lipid nanoparticle of any one of claims 1 to 30, wherein the ionizable lipid is DODAP, DODMA, DLinDMA, DLin-KC2-DMA, DLin-MC3-DMA, SM-102, or ALC-0315, or a combination thereof, or a stereoisomer, a mixture of stereoisomers, or a pharmaceutically acceptable salt thereof.

32. The lipid nanoparticle of any one of claims 1 to 31, wherein the molar percentage of ionizable lipid is 35-70 mol%, more preferably 40-60 mol%, such as 35 mol%, 40 mol%, 45 mol%, 50 mol%, 55 mol% or 60 mol%.

33. The lipid nanoparticle of any one of claims 1 to 32, further comprising a phospholipid.

34. The lipid nanoparticle of claim 33, wherein the phospholipid is DSPC, DMPC, DOPC, DPPC, POPC, DOPE, DMPE, POPOE, or DPPE.

35. The lipid nanoparticle according to any one of claims 1 to 34, wherein the molar percentage of phospholipids is 5-30 mol%, preferably 5-20 mol%, more preferably 5-15 mol%, such as 5 mol%, 6 mol%, 7 mol%, 8 mol%, 9 mol%, 10 mol%, 11 mol%, 12 mol%, 13 mol%, 14 mol% or 15 mol%.

36. The lipid nanoparticle of any one of claims 1 to 35, further comprising a structured lipid.

37. The lipid nanoparticle of claim 36, wherein the structured lipid is selected from the group consisting of cholesterol, campesterol, stigmasterol, sitosterol, brassicasterol, ergosterol, solanine, ursolic acid, α-tocopherol, β-sitosterol, avenasterol, and rapeseedsterol.

38. The lipid nanoparticle of any one of claims 1-37, wherein the molar percentage of the structural lipid is 10-80 mol%, more preferably 15-70 mol%, more preferably 30-50 mol%, more preferably 30-40 mol%, such as 30 mol%, 35 mol%, 40 mol%, 45 mol%, 50 mol%, 55 mol% or 60 mol%.

39. The lipid nanoparticle according to any one of claims 1 to 38, further comprising a polymer-conjugated lipid, such as a PEGylated lipid.

40. The lipid nanoparticle of claim 39, wherein the PEGylated lipid comprises a PEGylated portion having a molecular weight of about 1000 Da to about 20,000 Da, about 1000 Da to about 5000 Da, or about 1000 Da to about 2000 Da.

41. The lipid nanoparticle of claim 39, wherein the polymer-conjugated lipid is ALC-0159, DMG-PEG2000, DMPE-PEG1000, DPPE-PEG1000, DSPE-PEG1000, DOPE-PEG1000, ceramide-PEG2000, DMPE-PEG2000, DPPE-PEG2000, DSPE-PEG2000, DSPE-PEG2000-mannose, ceramide-PEG5000, DSPE-PEG5000, or DSPE-PEG2000 amine.

42. The lipid nanoparticle according to any one of claims 1 to 41, wherein the molar percentage of the polymer-conjugated lipid is 0.25-10 mol%, more preferably 0.5-5 mol%, more preferably 0.5-3 mol%, such as 0.5 mol%, 0.6 mol%, 0.7 mol%, 0.8 mol%, 0.9 mol%, 1.0 mol%, 1.1 mol%, 1.2 mol%, 1.3 mol%, 1.4 mol%, 1.5 mol%, 1.6 mol%, 1.7 mol%. l%, 1.8mol%, 1.9mol%, 2.0mol%, 2.1mol%, 2.2mol%, 2.3mol%, 2.4mol%, 2.5mol%, 2.6mol%, 2.7mol%, 2.8mol%, 2. 9mol%, 3.0mol%, 3.1mol%, 3.2mol%, 3.3mol%, 3.4mol%, 3.5mol%, 3.6mol%, 3.7mol%, 3.8mol%, 3.9mol%, 4.0mol%.

43. The lipid nanoparticle according to any one of claims 1-42, wherein the lipid nanoparticle contains: (1) a steroid compound; (2) an ionizable lipid; (3) a phospholipid compound; (4) a structural lipid; and (5) a polymer-conjugated lipid.

44. The lipid nanoparticle of any one of claims 1-43, wherein the spleen-targeted lipid nanoparticle comprises the following molar percentages of components: 1) 3-15 mol% of a steroid compound, preferably 5-10 mol%; 2) 35-70 mol% ionizable lipids, preferably 40-60 mol%; 3) 5-30 mol% of phospholipids, preferably 5-20 mol%; 4) structural lipids 10-80 mol%, preferably 15-70 mol%, more preferably 30-50 mol%; and 5) Polymer-conjugated lipid 0.25-10 mol%, preferably 0.5-5 mol%, more preferably 0.5-3 mol%.

45. The lipid nanoparticle of any one of claims 1 to 44, wherein the lipid nanoparticle further comprises a payload, and wherein the payload is a therapeutic, prophylactic, or diagnostic agent.

46. ​​The lipid nanoparticle of claim 45, wherein the payload is a nucleic acid.

47. The lipid nanoparticle of claim 46, wherein the nucleic acid is an antisense oligonucleotide (ASO), DNA, or RNA, optionally wherein the RNA is RNA interference (RNAi), small interfering RNA (siRNA), short hairpin RNA (shRNA), antisense RNA (aRNA), messenger RNA (mRNA), modified messenger RNA (mmRNA), long non-coding RNA (lncRNA), microRNA (miRNA), small activating RNA (saRNA), poly-encoded nucleic acid (MCNA), polymer-encoded nucleic acid (PCNA), guide RNA (gRNA), CRISPR RNA (crRNA), or any other RNA in a ribozyme.

48. A lipid nanoparticle composition comprising the lipid nanoparticles according to any one of claims 1 to 47.

49. A pharmaceutical composition comprising the lipid nanoparticle according to any one of claims 1 to 47 or the lipid nanoparticle composition according to claim 48 and a pharmaceutically acceptable carrier.

50. A method for treating or preventing a spleen disease or a spleen-related disease in a subject suffering from the spleen disease or the spleen-related disease, comprising administering a therapeutically effective amount of the lipid nanoparticle of any one of claims 1 to 47, the lipid nanoparticle composition of claim 48, or the pharmaceutical composition of claim 49 to the subject.

51. Use of the lipid nanoparticle of any one of claims 1 to 47 or the lipid nanoparticle composition of claim 48 in the preparation of a medicament for preventing or treating spleen diseases or spleen-related diseases.

52. The lipid nanoparticle of any one of claims 1 to 47, the lipid nanoparticle composition of claim 48, or the pharmaceutical composition of claim 49, for preventing or treating a spleen disease or a spleen-related disease; preferably, the spleen disease or spleen-related disease comprises a blood system disease, an infectious disease, an immune disease, or a cancer; more preferably, the spleen disease or spleen-related disease comprises hemolytic anemia, thrombocytopenic purpura, chronic leukemia, lymphoma, such as Hodgkin's lymphoma or non-Hodgkin's lymphoma, myelodysplastic syndrome , myelofibrosis, splenomegaly, lipid metabolism disorders such as glucocerebrosidosis and sphingomyelinosis, splenomegaly and hypersplenism associated with sepsis, typhoid fever, infectious mononucleosis or subacute bacterial endocarditis, splenomegaly and hypersplenism associated with malaria, tuberculosis, kala-azar, etc., congestive splenomegaly and hypersplenism caused by portal hypertension in cirrhosis, splenic cysts such as parasitic splenic cysts or pseudosplenic cysts, splenic abscesses, primary splenic tumors, splenic metastases, splenic artery aneurysms, splenic infarction, splenic purpura, splenic rupture, etc.

53. The method of claim 50 or the use of claim 51, wherein the spleen disease or spleen-related disease comprises a blood system disease, an infectious disease, an immune disease or cancer.

54. The method of claim 50 or the use of claim 51, wherein the spleen disease or spleen-related disease comprises hemolytic anemia, thrombocytopenic purpura, chronic leukemia, lymphoma, such as Hodgkin's lymphoma or non-Hodgkin's lymphoma, myelodysplastic syndrome, myelofibrosis, splenomegaly, lipid metabolism disorders, such as glucocerebrosidosis and sphingomyelinosis, splenomegaly and hypersplenism associated with sepsis, typhoid fever, infectious mononucleosis or subacute bacterial endocarditis, splenomegaly and hypersplenism associated with malaria, tuberculosis, kala-azar, etc., congestive splenomegaly and hypersplenism caused by portal hypertension in cirrhosis, splenic cysts, such as parasitic splenic cysts or pseudosplenic cysts, splenic abscesses, primary splenic tumors, splenic metastases, splenic artery aneurysms, splenic infarction, splenic purpura, splenic rupture, etc.

55. A method for preparing the lipid nanoparticle of any one of claims 1 to 47 or the lipid nanoparticle composition of claim 48, comprising the steps of: (i) dissolving a mixture comprising lipids in a first solution, wherein the lipid solution is formed in an organic solvent; (ii) dissolving the payload in a second solution to form a payload solution; as well as (iii) mixing the lipid solution and the payload solution to form the lipid nanoparticles.

56. The method of claim 55, wherein the mixing is performed using a microfluidic system.

57. A steroid compound of formula (II-A): or a stereoisomer, a mixture of stereoisomers, or a pharmaceutically acceptable salt thereof, wherein: Each are independently a single bond or a double bond; Y is -C(O)O-, -O-, -NH-, -SC(O)O-, -OC(O)NR 2a -、-NR 2a C(O)NR 2a -、-OC(O)S-、-OC(O)O-、-NR 2a C(O)O-, -OC(O)-, -SC(O)-, -C(O)S-, -NR 2a -、-C(O)NR 2a -、-NR 2a C(O)-、-NR 2a C(O)S-、-SC(O)NR 2a -, -C(O)-, -OC(S)-, -C(S)O-, -OC(S)NR 2a -、-NR 2a C(S)O- or -S(O) 0-2 -, and wherein the leftward connection is in the direction of V1; Y1 is absent, -C(O)O-, -O-, -NH-, -SC(O)O-, -OC(O)NR 2a -、-NR 2a C(O)NR 2a -、-OC(O)S-、-OC(O)O-、-NR 2a C(O)O-, -OC(O)-, -SC(O)-, -C(O)S-, -NR 2a -、-C(O)NR 2a -、-NR 2a C(O)-、-NR 2a C(O)S-、-SC(O)NR 2a -, -C(O)-, -OC(S)-, -C(S)O-, -OC(S)NR 2a -、-NR 2a C(S)O- or -S(O) 0-2 -, and wherein the leftward connection is in the direction of V1; R 2a Each instance of is independently H, C 1-10 Alkyl, C 3-14 cycloalkyl or 3 to 14 membered heterocyclyl, and wherein the alkyl, cycloalkyl and heterocyclyl are independently optionally replaced by C 1-10 Alkyl, L 2b -OR 2b , L b -SR 2b , L 2b -NR 2b R' 2b , L 2b N + R 2b R' 2b R” 2b , L 2b -OC(O)R 2b or L 2b -C(O)OR 2b replace; R 201 、R 202、 R 203 、R 204 Each example is independently H, OH, alkyl, alkoxy, -L 2b -NR 21 R 22 or -L 2b -N + R 21 R 22 R 23 , L 2b -OC(O)R 21 or L 2b -C(O)OR 21 ; m is an integer from 3 to 12; m1 is an integer from 0 to 12; V1 is NR 21 R 22 、N + R 21 R 22 R 23 , imidazole, pyridine, pyrrole, pyrrolidine, pyrazole, (iso)thiazole, (iso)oxazole, oxadiazine, oxazoline, dithiourea, (iso)triazole, tetrazole, pentazole, indole, indoline, pyrimidine, pyrazine, quinazoline, piperazine, piperidine, morpholine, pyran, quinoxaline, quinoline, quinolinone, (iso)quinoline, amidine, guanidine or urea; M is absent or one or more anions independently selected from chloride, bromide, iodide, sulfate, nitrate, perchlorate, formate, citrate, sulfonate, methanesulfonate, trifluoromethanesulfonate, trifluoroacetate, pyrosulfate, bisulfate, sulfite, bisulfite, phosphate, pyrophosphate, monohydrogen phosphate, dihydrogen phosphate, acetate, propionate, decanoate, octanoate, acrylate, isobutyrate, hexanoate, heptanoate, oxalate, malonate, succinate, fumarate, maleate, benzoate, phthalate, phenylacetate, lactate, glycolate, tartrate, mandelate, methanesulfonate, gluconate, aspartate, or glutamate; R 21 、R 22 、R 23 are each independently H, optionally substituted C 1-8 Alkyl, -L 2b -OR 2b 、-L 2b -SR 2b 、-L 2b -NR 2b R' 2b R” 2b , L 2b -OC(O)R 2b or L 2b -C(O)OR 2b ; L 2b is absent or optionally substituted C 1-10 alkylene; R 2b , R' 2b and R” 2b Each of is independently H or C 1-10 alkyl; and R z1 It is C 1-14 Alkyl or C 1-14 alkenyl, wherein the alkyl and alkenyl groups are optionally substituted; and with the proviso that the compound is not:

58. A steroid compound represented by formula (II-B): or a stereoisomer, a mixture of stereoisomers, or a pharmaceutically acceptable salt thereof, wherein: Y is -C(O)O-, -O-, -NH-, -SC(O)O-, -OC(O)NR 2a -、-NR 2a C(O)NR 2a -、-OC(O)S-、-OC(O)O-、-NR 2a C(O)O-, -OC(O)-, -SC(O)-, -C(O)S-, -NR 2a -、-C(O)NR 2a -、-NR 2a C(O)-、-NR 2a C(O)S-、-SC(O)NR 2a -, -C(O)-, -OC(S)-, -C(S)O-, -OC(S)NR 2a -、-NR 2a C(S)O- or -S(O) 0-2 -, and wherein the leftward connection is in the direction of V1; Y1 is absent, -C(O)O-, -O-, -NH-, -SC(O)O-, -OC(O)NR 2a -、-NR 2a C(O)NR 2a -、-OC(O)S-、-OC(O)O-、-NR 2a C(O)O-, -OC(O)-, -SC(O)-, -C(O)S-, -NR 2a -、-C(O)NR 2a -、-NR 2a C(O)-、-NR 2a C(O)S-、-SC(O)NR 2a -, -C(O)-, -OC(S)-, -C(S)O-, -OC(S)NR 2a -、-NR 2a C(S)O- or -S(O) 0-2 -, and wherein the leftward connection is in the direction of V1; R 2a Each instance of is independently H, C 1-10 Alkyl, C 3-14 cycloalkyl or 3 to 14 membered heterocyclyl, and wherein the alkyl, cycloalkyl and heterocyclyl are independently optionally replaced by C 1-10 Alkyl, L 2b -OR 2b , L b -SR 2b , L 2b -NR 2b R' 2b , L 2b N + R 2b R' 2b R” 2b , L 2b -OC(O)R 2b or L 2b -C(O)OR 2b replace; R 201 、R 202、 R 203 、R 204 Each example is independently H, OH, alkyl, alkoxy, -L 2b -NR 21 R 22 or -L 2b -N + R 21 R 22 R 23 , L 2b -OC(O)R 21 or L 2b -C(O)OR 21 ; m is an integer from 5 to 12; m1 is an integer from 0 to 12; V1 is NR 21 R 22 、N + R 21 R 22 R 23 , imidazole, pyridine, pyrrole, pyrrolidine, pyrazole, (iso)thiazole, (iso)oxazole, oxadiazine, oxazoline, dithioureaazole, (iso)triazole, tetrazole, pentazole, indole, indoline, pyrimidine, pyrazine, quinazoline, piperazine, piperidine, morpholine, pyran, quinoxaline, quinoline, quinolinone, (iso)quinoline, amidine, guanidine or urea; M is absent or is one or more anions independently selected from chloride, bromide, iodide, sulfate, nitrate, perchlorate, formate, citrate, sulfonate, methanesulfonate, trifluoromethanesulfonate, trifluoroacetate, pyrosulfate, bisulfate, sulfite, bisulfite, phosphate, pyrophosphate, monohydrogen phosphate, dihydrogen phosphate, acetate, propionate, decanoate, octanoate, acrylate, isobutyrate, hexanoate, heptanoate, oxalate, malonate, succinate, fumarate, maleate, benzoate, phthalate, phenylacetate, lactate, glycolate, tartrate, mandelate, methanesulfonate, gluconate, aspartate, or glutamate; R 21 、R 22 、R 23 are each independently H, optionally substituted C 1-8 Alkyl, -L 2b -OR 2b 、-L 2b -SR 2b 、-L 2b -NR 2b R' 2b R” 2b , L 2b -OC(O)R 2b or L 2b -C(O)OR 2b ; L 2b is absent or optionally substituted C 1-10 alkylene; R 2b , R' 2b and R” 2b Each of is independently H or C 1-10 alkyl; Preferably, Y1 is -C(O)O-, -OC(O)-, -O-, -S-, -NR 2a -、-OC(O)NR 2a -or-NR 2a C(O)-.

59. A steroid compound having the formula (IP): or a stereoisomer, a mixture of stereoisomers, or a pharmaceutically acceptable salt thereof, wherein: Is a single bond or a double bond; Y1 does not exist, -C(O)O-, -O-, -NH-, -SC(O)O-, -OC(O)NR 2a -、-NR 2a C(O)NR 2a -、-OC(O)S-、-OC(O)O-、-NR 2a C(O)O-, -OC(O)-, -SC(O)-, -C(O)S-,-NR 2a -,-C(O)NR 2a -,-NR 2a C(O)-,-NR 2a C(O)S-,-SC(O)NR 2a -,-C(O)-,-OC(S)-, -C(S)O-, -OC(S)NR 2a -、-NR 2a C(S)O-, -S(O) 0-2 -or -OPO3-, and wherein the leftward connection is in the direction of V1; R 2a Each instance of is independently H, C 1-10 Alkyl, C 3-14 cycloalkyl or 3 to 14 membered heterocyclyl, and wherein the alkyl, cycloalkyl and heterocyclyl are independently optionally replaced by C 1-10 Alkyl, L 2b -OR 2b , L b -SR 2b , L 2b -NR 2b R' 2b , L 2b N + R 2b R' 2b R” 2b , L 2b -OC(O)R 2b or L 2b -C(O)OR 2b replace; R 201 、R 202、 R 203 、R 204 Each example is independently H, OH, alkyl, alkoxy, -L 2b -NR 21 R 22 、-L 2b -N + R 21 R 22 R 23 , L 2b -OC(O)R 21 or L 2b -C(O)OR 21 ; R 205 Absence, cations, H, C 1-6 Alkyl, C 1-6 Halogenated alkyl, C 2-6 Alkenyl or C 2-6 alkynyl, and wherein alkyl, haloalkyl, alkenyl and alkynyl are optionally substituted; m is an integer from 1 to 12; m1 is an integer from 0 to 12; V1 is NR 21 R 22 、N + R 21 R 22 R 23 , imidazole, pyridine, pyrrole, pyrrolidine, pyrazole, (iso)thiazole, (iso)oxazole, oxadiazine, oxazoline, dithioureaazole, (iso)triazole, tetrazole, pentazole, indole, indoline, pyrimidine, pyrazine, quinazoline, piperazine, piperidine, morpholine, pyran, quinoxaline, quinoline, quinolinone, (iso)quinoline, amidine, guanidine or urea; M is absent or is one or more anions independently selected from chloride, bromide, iodide, sulfate, nitrate, perchlorate, formate, citrate, sulfonate, methanesulfonate, trifluoromethanesulfonate, trifluoroacetate, pyrosulfate, bisulfate, sulfite, bisulfite, phosphate, pyrophosphate, monohydrogen phosphate, dihydrogen phosphate, acetate, propionate, decanoate, octanoate, acrylate, isobutyrate, hexanoate, heptanoate, oxalate, malonate, succinate, fumarate, maleate, benzoate, phthalate, phenylacetate, lactate, glycolate, tartrate, mandelate, methanesulfonate, gluconate, aspartate, or glutamate; R 21 、R 22 、R 23 Each instance of is independently H, optionally substituted C 1-8 Alkyl, -L 2b -OR 2b 、-L 2b -SR 2b 、-L 2b -NR 2b R' 2b R” 2b , L 2b -OC(O)R 2b or L 2b -C(O)OR 2b ; L 2b is absent or optionally substituted C 1-10 alkylene; R 2b , R' 2b and R” 2b Each instance of is independently H or C 1-10 alkyl; R z1 It is C 1-14 Alkyl or C 1-14 alkenyl, wherein the alkyl and alkenyl groups are optionally substituted.

60. A steroid compound represented by formula (II-C1): or a stereoisomer, a mixture of stereoisomers, or a pharmaceutically acceptable salt thereof, wherein: R z1 yes R 2a is H, optionally substituted C 1-8 Alkyl, -L 2b -OR 2b , L 2b -OC(O)R 2b or L 2b -C(O)OR 2b , preferably H, -CH3, -CH2C(O)OCH3 or -CH2C(O)OH; V1 is NR 21 R 22 or N + R 21 R 22 R 23 ; where R 21 、R 22 、R 23 are each independently H, optionally substituted C 1-8 Alkyl, -L 2b -OR 2b 、-L 2b -SR 2b 、-L 2b -NR 2b R' 2b R” 2b , L 2b -OC(O)R 2b or L 2b -C(O)OR 2b ; and R 21 and R 22 Not simultaneously hydrogen; L 2b is absent or optionally substituted C 1-10 alkylene; R 2b , R' 2b and R” 2b Each of is independently H or C 1-10 alkyl; M is absent or is one or more anions independently selected from chloride, bromide, iodide, sulfate, nitrate, perchlorate, formate, citrate, sulfonate, methanesulfonate, trifluoromethanesulfonate, trifluoroacetate, pyrosulfate, bisulfate, sulfite, bisulfite, phosphate, pyrophosphate, monohydrogen phosphate, dihydrogen phosphate, acetate, propionate, decanoate, octanoate, acrylate, isobutyrate, hexanoate, heptanoate, oxalate, malonate, succinate, fumarate, maleate, benzoate, phthalate, phenylacetate, lactate, glycolate, tartrate, mandelate, methanesulfonate, gluconate, aspartate, or glutamate; m is an integer from 1 to 10, for example, 1, 2, 3, 4, 5, 6, 7, 8, 9 or 10.

61. A steroid compound represented by formula (II-C2): or a stereoisomer, a mixture of stereoisomers, or a pharmaceutically acceptable salt thereof, wherein: R z1 yes V1 is NR 21 R 22 or N + R 21 R 22 R 23 ; where R 21 、R 22 、R 23 are each independently H, optionally substituted C 1-8 Alkyl, -L 2b -OR 2b 、-L 2b -SR 2b 、-L 2b -NR 2b R' 2b R” 2b , L 2b -OC(O)R 2b or L 2b -C(O)OR 2b ; and R 21 and R 22 Not simultaneously hydrogen; L 2b is absent or optionally substituted C 1-10 alkylene; R 2b , R' 2b and R” 2b Each of is independently H or C 1-10 alkyl; M is absent or is one or more anions independently selected from chloride, bromide, iodide, sulfate, nitrate, perchlorate, formate, citrate, sulfonate, methanesulfonate, trifluoromethanesulfonate, trifluoroacetate, pyrosulfate, bisulfate, sulfite, bisulfite, phosphate, pyrophosphate, monohydrogen phosphate, dihydrogen phosphate, acetate, propionate, decanoate, octanoate, acrylate, isobutyrate, hexanoate, heptanoate, oxalate, malonate, succinate, fumarate, maleate, benzoate, phthalate, phenylacetate, lactate, glycolate, tartrate, mandelate, methanesulfonate, gluconate, aspartate, or glutamate; m is an integer from 1 to 10, for example, 1, 2, 3, 4, 5, 6, 7, 8, 9 or 10.

62. A steroid compound selected from the group consisting of: or a stereoisomer, a mixture of stereoisomers, or a pharmaceutically acceptable salt thereof.

63. A lipid nanoparticle comprising a compound according to any one of claims 57-62.

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