Cationic lipid compounds and composition and use thereof

Abstract

The present application provides new cationic lipid compounds and a composition containing the compounds. A nanoparticle composition contains new cationic lipid compounds and other lipids, such as phospholipids, structural lipids and PEG lipids. Additionally, the nanoparticle composition of the present application can be used to deliver a therapeutic agent and / or a prophylactic agent to mammalian cells or organs to, for example, regulate polypeptide, protein or gene expression.

Description

Cationic lipid compound and its composition and application

[0001] This application claims priority to and the benefits of Chinese Patent Application No. 2024103242951 filed with the State Intellectual Property Office of China on March 20, 2024, the disclosure of which is incorporated herein by reference in its entirety. Technical Field

[0002] The present invention relates to the field of biomedicine technology, and in particular to a cationic lipid compound, a preparation method and an application thereof. Background Art

[0003] Liposomal nanoparticles (LNPs), developed based on ionizable cationic lipids, are a proven and effective platform for nucleic acid and drug delivery. LNPs are composed of four main components: ionizable cationic lipids, cholesterol, auxiliary cationic lipids, and polyethylene glycol lipids. The design and synthesis of ionizable cationic lipids is one of the core technologies of LNPs, playing a key role in the efficient delivery of nucleic acids into cells and their release into the cytoplasm.

[0004] Nucleic acid drugs mainly include antisense nucleic acids, small interfering nucleic acids (siRNA), micronucleic acids, messenger nucleic acids (mRNA) and CRISPR systems. These nucleic acid drugs can inhibit, interfere with, regulate, insert and transform human genes through various mechanisms, thereby achieving the treatment of specific diseases. However, the delivery of nucleic acid drugs brings unique challenges due to their easy degradation and the requirement for efficient intracellular delivery. The combination of nanotechnology and nucleic acid drug delivery solves many of these challenges. In recent years, lipid nanoparticles have demonstrated their practicality as a delivery platform for mRNA vaccines and therapies.

[0005] The successful application of ionizable cationic lipid molecules has rapidly advanced the clinical application of nucleic acid drugs. Onpattro, the first siRNA drug, uses DLin-MC3-DMA as an ionizable liposome for delivery into liver cells for the treatment of nerve damage caused by transthyretin amyloidosis (hATTR, familial amyloid polyneuropathy). Moderna and BioNTech / Pfizer have developed two COVID-19 vaccines using SM-102 and ALC-0315, respectively.

[0006] In addition to nucleic acid drugs, some poorly soluble small molecule drugs can also be better delivered with the help of lipid nanoparticles, such as protein degradation targeting chimeras (PROTACs), which have received much attention in recent years. PROTACs are highly anticipated in anti-tumor treatment because they can achieve efficient degradation of previously difficult-to-drug targets. However, PROTACs often have the characteristics of poor water solubility and large molecular weight, making it difficult to cross the body and cell barriers. Lipid nanoparticles provide a new option for the efficient delivery of this type of molecules.

[0007] CD36 is a ubiquitous scavenger receptor in the human body, helping immune cells recognize and eliminate pathogens associated with long-chain fatty acids, oxidized lipids, advanced oxidation protein products, thrombospondins, and advanced glycation end products. CD36 is widely expressed in muscle cells, gastrointestinal tissue cells, dendritic cells (DCs), microvascular endothelial cells (MVECs), retinal epithelial cells, monocytes, adipocytes, platelets, intestinal epithelial cells, microglia, and podocytes. Specifically, in tumor tissue, CD36 is expressed in tumor cells, stromal cells, and immune cells, but expression levels vary across cell types and tumor stages. It is generally believed that CD36 is highly expressed in ovarian cancer, gastric cancer, glioblastoma, and oral squamous cell carcinoma, but is low in highly metastatic cancers. CD36 plays a regulatory role in tumor immune evasion, growth, and metastasis. In highly metastatic tumors, it promotes metastasis by inducing epithelial-mesenchymal transition (EMT) through the TGF-β signaling pathway. CD36 has a hairpin-shaped structure outside the cell membrane that is divided into two binding regions, called entrance 1 and entrance 2. Entrance 1 is the main binding region.

[0008] Sulfosuccinimidyl oleate (SSO) is the first discovered CD36 inhibitor. It can be precisely recognized by entrance1 and permanently bind to CD36 (Sulfo-N-succinimidyl oleate (SSO) inhibits fatty acid uptake and signaling for intracellular calcium via binding CD36 lysine 164: SSO also inhibits oxidized low density lipoprotein uptake by macrophages. J Biol Chem. 2013 May 31; 288(22): 15547-55. doi: 10.1074 / jbc.M113.473298). A study prepared a microemulsion of SSO and 0.5% methylcellulose and administered it orally to Balb / c mice at a dose of 50 mg / kg, demonstrating the safety of the design (Sulfosuccinimidyl oleatesodium is neuroprotective and alleviates stroke-induced neuroinflammation. JNeuroinflammation. 2017Dec 4; 14(1): 237. doi: 10.1186 / s12974-017-1010-7).

[0009] To date, a variety of ionizable cationic lipid molecules have been developed, and through structural modification and formulation optimization, a variety of delivery functions have been achieved. However, there is still a need for improved ionizable lipid molecules for nucleic acid delivery to enhance delivery efficiency and increase the expression of therapeutic proteins. Existing delivery systems have difficulty targeting cells and tissues outside the liver, which greatly limits their scope of application. Furthermore, there are currently no reports of ionizable cationic lipid molecules that target CD36. Summary of the Invention

[0010] In view of this, the present invention provides an ionizable cationic lipid targeting CD36 based on the structural characteristics of entrance 1, and assembles it into lipid nanoparticles to achieve the purpose of precisely delivering nucleic acids or other drugs to cells, tissues or tumor microenvironments containing CD36 receptors.

[0011] To achieve the above objectives, the present invention mainly provides the following technical solutions:

[0012] In one aspect, the present application provides a compound of formula (I):

[0013] Among them, R 1 independently selected from the group consisting of hydroxy-substituted alkyl, hydroxy-substituted alkenyl, and hydroxy-substituted alkynyl;

[0014] M 1 and M 2 are each independently selected from the group consisting of optionally substituted alkylene, optionally substituted alkenylene, and optionally substituted alkynylene;

[0015] L 1 and L 2 are each independently selected from the group consisting of -C(O)O-, -OC(O)-, -O-, -C(O)N(R)-, and -N(R)C(O)-, wherein each R is independently selected from the group consisting of: C 1-3 Alkyl, C 2-3 alkenyl and H;

[0016] M 3 and M 4 Each independently where R 2 Independently selected from linear C 4-10 Alkylene, straight chain C 4-10 Alkenylene and straight chain C 4-10 The group consisting of alkynylene, R 3 Independently selected from linear C 4-10 Alkyl, straight chain C 4-10 Alkenyl and straight chain C 4-10 A group consisting of alkynyl groups.

[0017] In some embodiments, wherein R 2 Independently selected from linear C 6-9 Alkylene, straight chain C 6-9 Alkenylene and straight chain C 6-9 The group consisting of alkynylene, R 3 Independently selected from linear C 6-9 Alkyl, straight chain C 6-9 Alkenyl and straight chain C 6-9 A group consisting of alkynyl groups.

[0018] In some embodiments, wherein R 2 Independently linear C 6-9 Alkylene or linear C 6-9 Alkenylene, R 3 Independently linear C 6-9 Alkyl or linear C 6-9 Alkenyl.

[0019] In some embodiments, wherein R 2 and R 3They are not unsaturated hydrocarbon groups at the same time.

[0020] In some embodiments, wherein M 3 and M 4 Each independently a linear C 10-20 Alkenyl or straight chain C 10-20 Alkynyl.

[0021] In some embodiments, wherein M 3 and M 4 Each independently a linear C 14-18 Alkenyl or straight chain C 14-18 Alkynyl.

[0022] In some embodiments, wherein M 3 and M 4 Each independently a linear C 18 Alkenyl or straight chain C 18 Alkynyl.

[0023] In some embodiments, wherein M 3 and M 4 For straight chain C 18 Alkenyl.

[0024] In some embodiments, wherein R 1 independently selected from hydroxy-substituted C 1-6 Alkyl, hydroxy substituted C 2-6 Alkenyl and hydroxy substituted C 2-6 A group consisting of alkynyl groups.

[0025] In some embodiments, wherein R 1 C containing hydroxyl substitution 1-6 alkyl.

[0026] In some embodiments, wherein R 1 Selected from the group consisting of -CH2OH, -(CH2)2OH, -(CH2)3OH, -(CH2)4OH, -(CH2)5OH and -(CH2)6OH.

[0027] In some embodiments, wherein M 1 and M 2 Each independently is an optionally substituted C 1-4 Alkylene, and when M 1 and M 2 When R is C2 alkylene 1 Not -(CH2)6OH.

[0028] In some embodiments, wherein M 1 and M 2 are each independently an optionally substituted C 3-4Alkylene.

[0029] In some embodiments, wherein the compound has formula (II):

[0030] Wherein, m is 2, 3, 4 or 5;

[0031] Each n is independently 3 or 4;

[0032] L 1 and L 2 each independently selected from the group consisting of -C(O)O-, -OC(O)-, -O-, -C(O)N(H)-, and -N(H)C(O)-;

[0033] M 3 and M 4 Each independently where R 2 Independently selected from linear C 4-10 Alkylene, straight chain C 4-10 Alkenylene and straight chain C 4-10 The group consisting of alkynylene, R 3 Independently selected from linear C 4-10 Alkyl, straight chain C 4-10 Alkenyl and straight chain C 4-10 A group consisting of alkynyl groups.

[0034] In some embodiments, wherein R 2 Independently selected from linear C 6-9 Alkylene, straight chain C 6-9 Alkenylene and straight chain C 6-9 The group consisting of alkynylene, R 3 Independently selected from linear C 6-9 Alkyl, straight chain C 6-9 Alkenyl and straight chain C 6-9 A group consisting of alkynyl groups.

[0035] In some embodiments, wherein R 2 Independently linear C 6-9 Alkylene or linear C 6-9 Alkenylene, R 3 Independently linear C 6-9 Alkyl or linear C 6-9 Alkenyl.

[0036] In some embodiments, wherein R 2 and R 3 They are not unsaturated hydrocarbon groups at the same time.

[0037] In some embodiments, wherein M 3 and M 4Each independently a linear C 10-20 Alkenyl or straight chain C 10-20 Alkynyl.

[0038] In some embodiments, wherein M 3 and M 4 Each independently a linear C 14-18 Alkenyl or straight chain C 14-18 Alkynyl.

[0039] In some embodiments, wherein M 3 and M 4 Each independently a linear C 18 Alkenyl or straight chain C 18 Alkynyl.

[0040] In some embodiments, wherein M 3 and M 4 For straight chain C 18 Alkenyl.

[0041] In some embodiments, wherein the compound has formula (III):

[0042] Wherein, m is 2, 3 or 4;

[0043] L 1 and L 2 each independently selected from the group consisting of -C(O)O-, -OC(O)-, -CO-, -C(O)N(H)-, and -N(H)C(O)-;

[0044] M 3 and M 4 Each independently where R 2 Independently linear C 4-10 Alkylene or linear C 4-10 Alkenylene, R 3 Independently linear C 6-9 Alkyl or linear C 4-10 Alkenyl.

[0045] In some embodiments, wherein R 2 Independently linear C 6-9 Alkylene or linear C 6-9 Alkenylene, R 3 Independently linear C 6-9 Alkyl or linear C 6-9 Alkenyl.

[0046] In some embodiments, wherein R 2 and R 3 They are not all unsaturated hydrocarbon groups.

[0047] In some embodiments, wherein R 2 and R 3 Each independently a linear C 6-9 Alkylene.

[0048] In some embodiments, wherein M 3 and M 4 For straight chain C 10-20 Alkenyl or straight chain C 10-20 Alkynyl.

[0049] In some embodiments, wherein M 3 and M 4 For straight chain C 14-18 Alkenyl or straight chain C 14-18 Alkynyl.

[0050] In some embodiments, wherein M 3 and M 4 For straight chain C 18 Alkenyl.

[0051] In some embodiments, wherein M 3 and M 4 Each independently selected from:

[0052] In some embodiments, wherein M 3 and M 4 for

[0053] On the other hand, the present application provides a compound of formula (IV):

[0054] Wherein, m is 2 or 3;

[0055] L 1 and L 2 are each independently selected from the group consisting of -C(O)O-, -OC(O)-, -CO-, -C(O)N(H)- and -N(H)C(O)-.

[0056] In some embodiments, wherein L 1 and L 2 Each is independently selected from -C(O)O-, -OC(O)-, -O-, -C(O)N(H)- and -N(H)C(O)-.

[0057] In some embodiments, wherein L 1 and L 2 It is -C(O)O-.

[0058] In some embodiments, wherein L 1 and L 2 It is -OC(O)-.

[0059] In some embodiments, wherein L 1 and L 2 For -CO-.

[0060] In some embodiments, wherein L 1 and L 2 It is -C(O)N(H)-.

[0061] In some embodiments, wherein L 1 and L 2 It is -N(H)C(O)-.

[0062] On the other hand, the present application provides a compound selected from:

[0063] In another aspect, the present application provides a nanoparticle composition comprising a lipid component of the compound described herein.

[0064] In some embodiments, the lipid component further comprises phospholipids.

[0065] In some embodiments, the phospholipid is selected from the group consisting of 1,2-dilinoleoyl-sn-glycero-3-phosphocholine (DLPC), 1,2-dimyristoyl-sn-glycero-phosphocholine (DMPC), 1,2-dioleoyl-sn-glycero-3-phosphocholine (DOPC), 1,2-dipalmitoyl-sn-glycero-3-phosphocholine (DPPC), 1,2-distearoyl-sn-glycero-3-phosphocholine (DSPC), 1,2-diondecanoyl-sn-glycero-phosphocholine (DUPC), 1-palmitoyl-2-oleoyl-sn-glycero-3-phosphocholine (POPC), 1,2-di-O-octadecenyl-sn-glycero-3-phosphocholine (18:0Diether PC), 1-oleoyl-2-cholesteryl hemisuccinyl-sn-glycero-3-phosphocholine (OChemsPC), 1-hexadecyl-sn-glycero-3-phosphocholine (C16 Lyso PC), 1,2-dialinolenoyl-sn-glycero-3-phosphocholine, 1,2-diarachidonoyl-sn-glycero-3-phosphocholine, 1,2-didocosahexaenoyl-sn-glycero-3-phosphocholine, 1,2-dioleoyl-sn-glycero-3-phosphoethanolamine (DOPE), 1,2-diphytanoyl-sn-glycero-3-phosphoethanolamine (ME 16.0PE), 1,2-distearoyl-sn-glycero-3-phosphoethanolamine, 1,2-dilinoleoyl-sn-glycero-3-phosphoethanolamine, 1,2-dilinolenoyl-sn-glycero-3-phosphoethanolamine, 1,2-diarachidonoyl-sn-glycero-3-phosphoethanolamine, 1,2-bisdocosahexaenoyl-sn-glycero-3-phosphoethanolamine, 1,2-dioleoyl-sn-glycero-3-phospho-rac-(1-glycerol) sodium salt (DOPG), dipalmitoylphosphatidylglycerol (DPPG), palmitoyloleoyl phosphatidylethanolamine (POPE), distearoyl-phosphatidyl-ethanolamine (DSPE), dipalmitoylphosphatidylethanolamine (DPPE), dimyristoylphosphoethanolamine (DMPE), 1-stearoyl-2-oleoyl-stearoylethanolamine (SOPE), 1-stearoyl-2-oleoyl-phosphatidylcholine (SOPC), sphingomyelin, phosphatidylcholine, phosphatidylethanolamine, phosphatidylserine, phosphatidylinositol, phosphatidic acid, palmitoyloleoylphosphatidylcholine, lysophosphatidylcholine, lysophosphatidylethanolamine (LPE), and mixtures thereof.

[0066] In some embodiments, the phospholipid is DOPE.

[0067] In some embodiments, the phospholipid is DSPC.

[0068] In some embodiments, the lipid component further comprises a structural lipid.

[0069] In some embodiments, the structured lipid is selected from the group consisting of cholesterol, coprosterol, sitosterol, ergosterol, campesterol, stigmasterol, brassicasterol, tomatidine, ursolic acid, α-tocopherol, and mixtures thereof.

[0070] In some embodiments, the structured lipid is cholesterol.

[0071] In some embodiments, the lipid component further comprises PEG lipids.

[0072] In some embodiments, the PEG lipid is selected from the group consisting of PEG-modified phosphatidylethanolamine, PEG-modified phosphatidic acid, PEG-modified ceramide, PEG-modified dialkylamine, PEG-modified diacylglycerol, PEG-modified dialkylglycerol, and mixtures thereof.

[0073] In some embodiments, the PEG lipid comprises a PEG moiety having a size of about 1000 Da to about 20 kDa.

[0074] In some embodiments, the PEG lipid is selected from any one or a combination of more than one of PEG1000-DMG, PEG5000-DMG, PEG2000-DMG and PEG2000-DSPE.

[0075] In some embodiments, the PEG lipid is PEG2000-DMG.

[0076] In some embodiments, the lipid component further comprises phospholipids, structural lipids and PEG lipids.

[0077] In some embodiments, the lipid component comprises about 30 mol% to about 60 mol% of the compound, about 0 mol% to about 30 mol% of phospholipids, about 18.5 mol% to about 48.5 mol% of structural lipids, and about 0 mol% to about 10 mol% of PEG lipids.

[0078] In some embodiments, the lipid component comprises about 50 mol% of the compound, about 10 mol% of phospholipids, about 38.5 mol% of structural lipids, and about 1.5 mol% of PEG lipids.

[0079] In some embodiments, the lipid component comprises about 50 mol% of Compound 1 or Compound 2, about 10 mol% of DOPE or DSPC, about 37.0-39.5 mol% of cholesterol or sitosterol, and about 0.5-3.0 mol% of PEG2000-DMG.

[0080] In some embodiments, the nanoparticle composition further comprises a therapeutic and / or prophylactic agent.

[0081] In some embodiments, the therapeutic and / or preventive agent is an anticancer agent, an antiviral agent, an immunomodulatory agent, an anti-inflammatory agent, or an agent that regulates cellular metabolic activity.

[0082] In some embodiments, the therapeutic and / or prophylactic agent is a nucleic acid, a protein, a peptide, or a small molecule.

[0083] In some embodiments, the therapeutic and / or prophylactic agent is a nucleic acid.

[0084] In some embodiments, the therapeutic and / or prophylactic agent is ribonucleic acid (RNA).

[0085] In some embodiments, the RNA is selected from the group consisting of small interfering RNA (siRNA), asymmetric interfering RNA (aiRNA), microRNA (miRNA), Dicer-substrate RNA (dsRNA), small hairpin RNA (shRNA), messenger RNA (mRNA), and mixtures thereof.

[0086] In some embodiments, the RNA is mRNA.

[0087] In some embodiments, the mRNA comprises one or more of the following: a stem-loop, a chain-terminating nucleoside, a poly(A) sequence, a polyadenylation signal, and / or a 5' cap structure.

[0088] In some embodiments, the therapeutic and / or prophylactic agent comprises components of a gene editing system.

[0089] In some embodiments, the components of the gene editing system comprise a polynucleotide encoding a nuclease.

[0090] In some embodiments, the nuclease is selected from the group consisting of zinc finger nucleases (ZFNs), transcription activator-like effector nucleases (TALENs), CRISPR-associated protein 9 (Cas9), and engineered homing endonucleases.

[0091] In some embodiments, wherein the nuclease is Cas9 and the nanoparticle composition further comprises a guide RNA that targets the nuclease to a specific site in the target cell genome.

[0092] In some embodiments, the components of the gene editing system include: i) CRISPR-associated protein 9 (Cas9), or mRNA encoding CRISPR-associated protein 9 (Cas9); and

[0093] ii) a guide RNA nucleic acid of a single guide RNA (sgRNA), or a guide RNA nucleic acid encoding a single guide RNA (sgRNA).

[0094] In some embodiments, the therapeutic and / or prophylactic agent comprises a small molecule.

[0095] In some embodiments, the therapeutic and / or prophylactic agent comprises a PROTAC (Proteolysis Targeting Chimera) molecule.

[0096] In some embodiments, the PROTAC is selected from the group consisting of ARV-110, ARV-471, ARV-766, ARV-771, AVR-825, AR-LDD, DT2216, KT-474, KT-413, KT-333, NX-2127, NX-5948, CG001419, CFT8634, FHD-609 and SARD279.

[0097] In some embodiments, the wt / wt ratio of the lipid component to the therapeutic and / or prophylactic agent is from about 10:1 to about 60:1.

[0098] In some embodiments, the nanoparticle composition has an N:P ratio of about 2:1 to about 30:1.

[0099] In some embodiments, the nanoparticle composition has an average size of about 70 nm to about 100 nm.

[0100] In another aspect, the present application provides use of the compound described herein and the nanoparticle composition described herein in preparing a delivery system.

[0101] In some embodiments, the delivery system is a CD36-targeted delivery system.

[0102] In another aspect, the present application provides a delivery system comprising the compound described herein or the nanoparticle composition described herein.

[0103] In some embodiments, the delivery system is a CD36-targeted delivery system.

[0104] On the other hand, the present application provides a pharmaceutical composition comprising the nanoparticle composition described herein and a pharmaceutically acceptable carrier.

[0105] In another aspect, the present application provides a method for delivering a therapeutic and / or prophylactic agent to a mammalian cell, the method comprising administering the nanoparticle composition described herein to a subject in need thereof, wherein the administration comprises contacting the cell with the nanoparticle composition, thereby delivering the therapeutic and / or prophylactic agent to the cell.

[0106] In some embodiments, the cells express CD36.

[0107] In some embodiments, the mammalian cell is in a mammal.

[0108] In some embodiments, the mammal is a human.

[0109] In another aspect, the present application provides a method for delivering a therapeutic agent and / or a prophylactic agent to an organ or tissue, the method comprising contacting the organ or tissue with the nanoparticle composition described herein, thereby delivering the therapeutic agent and / or prophylactic agent to the cell.

[0110] In some embodiments, the organ or tissue is an organ or tissue that expresses CD36.

[0111] In another aspect, the present application provides a method for preventing and / or treating a disease or condition, comprising administering an effective amount of the nanoparticle composition described herein to a subject in need thereof. BRIEF DESCRIPTION OF THE DRAWINGS

[0112] The accompanying drawings, which are incorporated into and constitute part of the specification, illustrate several embodiments of the present application and, together with the specification, serve to explain the principles of the present application. The accompanying drawings are only for the purpose of illustrating the embodiments of the present application and should not be construed as limiting the present application. Further objects, features, and advantages of the present application will become apparent from the following detailed description taken in conjunction with the accompanying drawings, which show illustrative embodiments of the present application, in which:

[0113] FIG1 shows the fluorescence photography of DC2.4 cells after transfection of the liposome nanoparticles LNP001 of the present application;

[0114] FIG2 shows the fluorescence photography of A549 cells after transfection of the liposome nanoparticles LNP001 of the present application;

[0115] FIG3A shows the fluorescence expression of the liposome nanoparticles LNP001 and LNP002 of the present application after intramuscular injection;

[0116] FIG3B shows the fluorescence photography of the liposome nanoparticles LNP001 and LNP002 after intravenous injection of the present application;

[0117] FIG3C shows the fluorescence photography of the liposome nanoparticles LNP001 and LNP002 after subcutaneous injection of the present application;

[0118] FIG4 shows the delivery results of the muscle-targeted gene editing system of the liposome nanoparticle LNP001 of the present application;

[0119] FIG5 shows the PROTAC drug delivery results of the liposome nanoparticle LNP001 of the present application targeting Hela cells. DETAILED DESCRIPTION

[0120] Definition of terms

[0121] In this application, the terms "alkyl" and "alkylene" generally refer to a straight-chain or branched saturated hydrocarbon moiety. In one embodiment, the alkyl group is a straight-chain saturated hydrocarbon. Unless otherwise indicated, the "alkyl" or "alkylene" group contains 1-24 carbon atoms. Representative saturated straight-chain alkyl groups include methyl, ethyl, n-propyl, n-butyl, n-pentyl, and n-hexyl. Representative saturated branched alkyl groups include isopropyl, sec-butyl, isobutyl, tert-butyl, and isopentyl. The symbol "C 1-14 "Alkyl" means an optionally substituted straight or branched chain saturated hydrocarbon containing 1 to 14 carbon atoms. Unless otherwise specified, the alkyl groups described herein refer to both unsubstituted and substituted alkyl groups.

[0122] In this application, the terms "alkenyl" and "alkenylene" generally refer to a straight or branched hydrocarbon moiety having one or more carbon-carbon double bonds. In one embodiment, the alkenyl group contains 1, 2, or 3 double bonds and is unsaturated. Unless otherwise indicated, an "alkenyl" group contains 2-24 carbon atoms. Alkenyl groups include cis and trans isomers. Representative straight and branched alkenyl groups include ethenyl, propenyl, 1-butenyl, 2-butenyl, isobutenyl, 1-pentenyl, 2-pentenyl, 3-methyl-1-butenyl, 2-methyl-2-butenyl, and 2,3-dimethyl-2-butenyl. The symbol "C 2-14 "Alkenyl" means an optionally substituted straight or branched chain hydrocarbon containing 2 to 14 carbon atoms and at least one carbon-carbon double bond. An alkenyl group may contain one, two, three, four or more carbon-carbon double bonds. For example, C 18 Alkenyl groups may include one or more double bonds, including two double bonds of C 18Unless otherwise specified, the alkenyl groups described herein refer to both unsubstituted and substituted alkenyl groups.

[0123] In this application, the terms "alkynyl" and "alkynylene" generally refer to straight or branched chain hydrocarbon moieties having one or more carbon-carbon triple bonds. Unless otherwise indicated, "alkynyl" contains 2-24 carbon atoms. Representative straight and branched chain alkynyl groups include ethynyl, propynyl, 1-butynyl, 2-butynyl, 1-pentynyl, 2-pentynyl, and 3-methyl-1-butynyl. The symbol "C 2-14 "Alkynyl" means an optionally substituted straight or branched chain hydrocarbon containing 2 to 14 carbon atoms and at least one carbon-carbon triple bond. Alkynyl groups may contain one, two, three, four or more carbon-carbon triple bonds. For example, C 18 Alkynyl groups may include one or more carbon-carbon triple bonds. Unless otherwise specifically stated, alkynyl groups described herein refer to both unsubstituted and substituted alkynyl groups.

[0124] In this application, the terms "optionally substituted alkyl", "optionally substituted alkenyl" and "optionally substituted alkynyl" generally mean that when substituted, at least one hydrogen atom is replaced by a substituent. Substituents include bridge oxygen, halogen, heterocycle, -CN, -OR x 、-NR x R y 、-NR x C(=O)R y 、-NR x SO2R y 、-C(=O)R x 、-C(=O)OR x 、-C(=O)NR x R y 、-SO n R x and-SO n NR x R y , where n is 0, 1 or 2, R x and R y are the same or different and are independently halogen, alkyl or heterocycle, and each of the alkyl and heterocycle substituents may be bridged by one or more oxygen, halogen, -OH, -CN, alkyl, -OR x , heterocyclic, -NR x R y 、-NR x C(=O)R y -NR x SO2R y 、-C(=O)R x 、-C(=O)OR x 、-C(=O)NRx R y 、-SO n R x and-SO n NR x R y Further replacement.

[0125] In this application, the term "compound" is intended to include all isomers and isotopes of the depicted structure. "Isotopes" refer to atoms with the same atomic number but different mass numbers due to the number of neutrons in the nucleus. For example, isotopes of hydrogen include tritium and deuterium.

[0126] In the present application, the term "isomer" generally refers to any geometric isomer, tautomer, zwitterion, stereoisomer, enantiomer or diastereomer of a compound. A compound may include one or more chiral centers and / or double bonds, and therefore may exist as a stereoisomer, such as a double bond isomer (i.e., geometric E / Z isomer) or a diastereomer (e.g., enantiomer (i.e., (+) or (-)) or cis / trans isomer). The present application encompasses any and all isomers of compounds described herein, including stereoisomerically pure forms (e.g., geometrically pure, enantiomerically pure or diastereomerically pure) and enantiomers and stereoisomer mixtures, such as racemates. Enantiomers and stereoisomer mixtures of compounds and the manner in which they are split into their constituent enantiomers or stereoisomers are well known.

[0127] In this application, "nanoparticle composition" generally refers to a composition comprising one or more lipids. Nanoparticle compositions are typically about micron-sized or smaller and can include a lipid bilayer. Nanoparticle compositions encompass lipid nanoparticles (LNPs), liposomes (e.g., lipid vesicles), and lipid complexes (e.g., cationic lipid complexes (lipoplex, LPX)). For example, a nanoparticle composition can be a liposome with a lipid bilayer having a diameter of 500 nm or less.

[0128] In this application, "lipid component" generally refers to a component of a nanoparticle composition that includes one or more lipids. For example, the lipid component can include one or more cationic / ionizable lipids, pegylated lipids, structural lipids, or other lipids, such as phospholipids.

[0129] In this application, the term "phospholipid" generally refers to a lipid comprising a phosphate moiety and one or more carbon chains, such as unsaturated fatty acid chains. A phospholipid can include one or more multiple bonds (e.g., double bonds or triple bonds) (e.g., one or more unsaturations). Specific phospholipids can contribute to membrane fusion. For example, a cationic phospholipid can interact with one or more negatively charged phospholipids of a membrane (e.g., a cell membrane or an intracellular membrane). Phospholipid fusion with the membrane can allow one or more components of a lipid-containing composition to pass through the membrane, thereby allowing, for example, the one or more components to be delivered to a cell.

[0130] In this application, "N:P ratio" generally refers to the molar ratio of ionizable (at physiological pH) nitrogen atoms in a lipid to phosphate groups in an RNA, such as in a nanoparticle composition comprising a lipid component and RNA.

[0131] In this application, the term "PEG lipid," "PEGylated lipid," or "PEGylated lipid" generally refers to a lipid that comprises a polyethylene glycol component.

[0132] The term "nucleic acid molecule" or "nucleic acid" or "polynucleotide" includes any compound and / or substance that comprises a nucleotide polymer. Each nucleotide is composed of a base, particularly a purine or pyrimidine base (i.e., cytosine (C), guanine (G), adenine (A), thymine (T) or uracil (U)), a sugar (i.e., deoxyribose or ribose) and a phosphate group. Typically, nucleic acid molecules are described by a base sequence, wherein the bases represent the primary structure (linear structure) of the nucleic acid molecule. The base sequence is typically expressed from 5' to 3'. The nucleic acids present in the nanoparticles according to the present invention include any known form of nucleic acid. In this article, the term nucleic acid molecule encompasses deoxyribonucleic acid (DNA) (including, for example, complementary DNA (cDNA) and genomic DNA), ribonucleic acid (RNA) (particularly messenger RNA (mRNA)), synthetic forms of DNA or RNA, and mixed polymers comprising two or more of these molecules. Nucleic acid molecules can be linear, hairpin-shaped or circular. In addition, the term nucleic acid molecule includes sense and antisense strands, as well as single-stranded and double-stranded forms. In addition, the nucleic acid molecules described herein may contain naturally occurring or non-naturally occurring nucleotides. Examples of non-naturally occurring nucleotides include modified nucleotide bases having derivatized sugar or phosphate backbone linkages or chemically modified residues.

[0133] In this application, the term "RNA" generally refers to ribonucleic acid that may be naturally occurring or non-naturally occurring. For example, RNA can include modified and / or non-naturally occurring components, such as one or more nucleobases, nucleosides, nucleotides, or linkers. RNA can include a cap structure, a chain-terminating nucleoside, a stem-loop, a polyadenylate sequence, and / or a polyadenylation signal. RNA can have a nucleotide sequence that encodes a polypeptide of interest. For example, RNA can be messenger RNA (mRNA). Translation of an mRNA encoding a specific polypeptide, for example, translation of an mRNA in vivo within a mammalian cell can produce the encoded polypeptide. RNA can be selected from the non-limiting group consisting of small interfering RNA (siRNA), asymmetric interfering RNA (aiRNA), microRNA (miRNA), Dicer-substrate RNA (dsRNA), small hairpin RNA (shRNA), mRNA, single-stranded guide RNA (sgRNA), cas9 mRNA, and mixtures thereof.

[0134] In this application, the term "contact" generally refers to establishing a physical connection between two or more entities. For example, contacting a mammalian cell with a nanoparticle composition means that the mammalian cell and the nanoparticle are physically connected. Methods for contacting cells with external entities in vivo and in vitro are well-known in the biological field. For example, contacting a nanoparticle composition with a mammalian cell in a mammal can be carried out by different routes of administration (e.g., intravenous, intramuscular, intradermal, and subcutaneous) and can involve different amounts of nanoparticle compositions. In addition, the nanoparticle composition can contact more than one mammalian cell.

[0135] In this application, the term "delivery" generally refers to providing an entity to a target. For example, delivering a therapeutic and / or prophylactic agent to a subject can involve administering a nanoparticle composition comprising the therapeutic and / or prophylactic agent to the subject (e.g., via an intravenous, intramuscular, intradermal, or subcutaneous route). Administering a nanoparticle composition to a mammal or mammalian cell can involve contacting one or more cells with the nanoparticle composition.

[0136] In this application, the term "enhanced delivery" means that the delivery of a therapeutic and / or prophylactic agent to a target tissue of interest (e.g., mammalian liver) by nanoparticles is higher (e.g., at least 1.5 times higher, at least 2 times higher, at least 3 times higher, at least 4 times higher, at least 5 times higher, at least 6 times higher, at least 7 times higher, at least 8 times higher, at least 9 times higher, at least 10 times higher) than the level of delivery of a therapeutic and / or prophylactic agent to a target tissue of interest (e.g., MC3, KC2, or DLinDMA) by control nanoparticles. The level of delivery of nanoparticles in a specific tissue can be measured by comparing the amount of protein produced in the tissue to the weight of the tissue, the amount of therapeutic and / or prophylactic agent in the tissue to the weight of the tissue, the amount of protein produced in the tissue to the amount of total protein in the tissue, or the amount of therapeutic and / or prophylactic agent in the tissue to the amount of total therapeutic and / or prophylactic agent in the tissue. It will be appreciated that it is not necessary to determine the enhanced delivery of nanoparticles to target tissues in a treated subject, as delivery can be determined in surrogates such as animal models (e.g., rat models). In certain embodiments, nanoparticle compositions comprising compounds according to Formula (I), (II), or (III) have substantially the same level of delivery enhancement regardless of the route of administration. For example, certain compounds disclosed herein exhibit similar delivery enhancement when used for intravenous or intramuscular delivery of therapeutic and / or prophylactic agents.

[0137] As used herein, the term "specific delivery," "specifically deliver," or "specifically delivering" means that delivery of a therapeutic and / or prophylactic agent to a target tissue of interest (e.g., expressing CD36) by nanoparticles is higher (e.g., at least 1.5-fold higher, at least 2-fold higher, at least 3-fold higher, at least 4-fold higher, at least 5-fold higher, at least 6-fold higher, at least 7-fold higher, at least 8-fold higher, at least 9-fold higher, at least 10-fold higher) than delivery to a non-target tissue (e.g., not expressing CD36). The level of nanoparticle delivery in a specific tissue can be measured by comparing the amount of protein produced in the tissue to the weight of the tissue, the amount of therapeutic and / or prophylactic agent in the tissue to the weight of the tissue, the amount of protein produced in the tissue to the total amount of protein in the tissue, or the amount of therapeutic and / or prophylactic agent in the tissue to the total amount of therapeutic and / or prophylactic agent in the tissue. For example, for renal vascular targeting, if, following systemic administration of the therapeutic and / or prophylactic agent, the therapeutic and / or prophylactic dose delivered to the kidney per 1 g of tissue is 1.5 times, 2 times, 3 times, 5 times, 10 times, 15 times, or 20 times the therapeutic and / or prophylactic dose delivered to the liver or spleen, then the therapeutic and / or prophylactic agent is specifically provided to the kidney of a mammal relative to the liver and spleen. It will be appreciated that specific delivery of nanoparticles to target tissues need not be determined in vivo in a treated subject and that delivery can be determined in surrogates such as animal models (e.g., rat models).

[0138] In the present application, the term "high expression" refers to that the expression level of a gene (such as CD36) in a specific cell or tissue is higher than the normal expression level of the gene (for example, at least 10% higher, preferably at least 25% higher, more preferably at least 50% higher, even more preferably at least 100% higher, even more preferably at least 200% higher, and most preferably at least 300% higher).

[0139] In this application, the term "in vitro" refers to events that occur in an artificial environment, such as in a test tube or reaction vessel, in a cell culture, in a Petri dish, etc., rather than inside an organism (such as an animal, plant, or microorganism).

[0140] In this application, the term "in vivo" refers to events that occur inside an organism (such as an animal, plant or microorganism, or a cell or tissue thereof).

[0141] In this application, the term "ex vivo" refers to an event that occurs outside an organism (e.g., an animal, plant, or microorganism, or a cell or tissue thereof). An ex vivo event can occur in an environment that is minimally altered relative to the natural (e.g., in vivo) environment.

[0142] In this application, "administration method" may include intravenous, intramuscular, intradermal, subcutaneous or other methods of delivering the composition to the subject. The choice of administration method should target delivery (eg, specific delivery) to a specific area or system of the body.

[0143] In this application, the term "pharmaceutically acceptable" is used herein to refer to compounds, materials, compositions and / or dosage forms that are, within the scope of sound medical judgment, suitable for use in contact with human and animal tissues without excessive toxicity, irritation, allergic response or other problems or complications, and are consistent with a reasonable benefit / risk ratio.

[0144] In this application, the term "pharmaceutically acceptable carrier" generally refers to ingredients other than active ingredients that are substantially non-toxic and non-inflammatory in the subject (eg, a vehicle capable of suspending, forming a complex, or dissolving the active compound).

[0145] In this application, "target cells" or "target cells" refer to one or more cells of interest (e.g., cells expressing or overexpressing CD36). These cells can be found in vitro, in vivo, in situ, or in a tissue or organ of an organism. The organism can be an animal, such as a mammal, and more specifically, a human.

[0146] In the present application, "target tissue" or "target tissue" refers to any one or more tissue types of interest to which the delivery of therapeutic and / or preventive agents will cause a desired biological and / or pharmacological effect. Examples of target tissues of interest include specific tissues, organs, and systems or groups thereof. In specific applications, the target tissue can be the vascular endothelium in the kidney, lung, spleen, blood vessels (intracoronary or femoral artery), or tumor tissue (e.g., by intratumoral injection). "Non-target tissue" refers to any one or more tissue types to which the expression of the encoded protein will not cause a desired biological and / or pharmacological effect. In specific applications, non-target tissues can include the liver and spleen.

[0147] In this application, the term "treat" refers to partially or completely alleviating, ameliorating, improving, alleviating a particular infection, disease, condition and / or illness, delaying its onset, inhibiting its progression, reducing its severity, and / or reducing the incidence of one or more symptoms or characteristics thereof. For example, "treating" cancer may refer to inhibiting the survival, growth and / or spread of a tumor. For the purpose of reducing the risk of developing pathology associated with a disease, condition and / or illness, treatment may be applied to a subject who does not exhibit symptoms of the disease, condition and / or illness, and / or to a subject who only exhibits early signs of the disease, condition and / or illness.

[0148] In this application, the terms "preventing," "prevent," or "protecting against" describe reducing or eliminating the onset of a disease, condition, or disorder, or reducing or eliminating the onset of symptoms or complications of the disease, condition, or disorder. For example, "prevention" can be with the aid of a vaccine, whereby the vaccine can be used to prevent a disease, condition, or disorder, such as preventing a viral infection.

[0149] Unless otherwise specified, an "individual" or "subject" is a human. In some cases, where specified, an "individual" or "subject" is or includes a non-human mammal (e.g., a "mammalian subject" or "non-human mammalian subject"). Mammals include, but are not limited to, domesticated animals (e.g., cattle, sheep, cats, dogs, and horses), primates (e.g., humans and non-human primates such as monkeys), rabbits, and rodents (e.g., mice and rats).

[0150] In this application, the terms "comprising" or "including" are used interchangeably, and are intended to be open and allow but not require the inclusion of other elements or steps. When the term "comprising" is used in this article, the terms "mainly consisting of" and "consisting of" are also encompassed and disclosed. Throughout the specification, when a composition is described as having, including or comprising a specific component, it is expected that the composition also mainly consists of or consists of said component. Similarly, when a method or process is described as having, including or comprising a specific process step, these processes also mainly consist of or consist of said process step. In addition, it should be understood that the order of the steps or the order in which certain operations are performed is unimportant, as long as the present invention remains operable. In addition, two or more steps or operations can be performed simultaneously.

[0151] In this application, whether or not explicitly stated, the term "about" refers to an index value, including, for example, integers, fractions, and percentages. The term "about" generally refers to a numerical range (e.g., + / - 5%-10% of the range) that one of ordinary skill in the art would consider to be equivalent to the value (e.g., having the same function or result). When terms such as "at least" and "about" appear before a numerical value or range list, the terms modify all values ​​or ranges provided in the list. In some cases, the term "about" can include numerical values ​​rounded to the nearest significant figure.

[0152] Unless otherwise indicated or apparent from the context, in the claims, words such as "a (kind)" and "said" may represent one (kind) or more than one (kind). Unless otherwise indicated or apparent from the context, a claim or description including "or" between one or more members of a group is deemed to satisfy that one, more than one or all of the group members are present in, used for, or otherwise relate to a given product or method. The present application includes embodiments in which only one member of the group is present in, used for, or otherwise relate to a given product or method. The present application includes embodiments in which more than one or all group members are present in, used for, or otherwise relate to a given product or method. Unless otherwise noted, as used herein, expressions such as "one or more of A, B, or C," "one or more A, B, or C," "one or more of A, B, and C," "one or more A, B, and C," "selected from A, B, and C," "selected from the group consisting of A, B, and C," etc. are used interchangeably and all refer to selecting from the group consisting of A, B, and / or C, i.e., one or more A, one or more B, one or more C, or any combination thereof.

[0153] Detailed Description of the Invention

[0154] Cationic lipid compounds

[0155] The present invention provides a series of ionizable cationic lipid molecules containing one or more double bonds of long-chain olefins, oleic acid or oleyl alcohol esters and their analogs. The central amine portion of the lipid according to formula (I), (II), (III) or (IV) can be protonated at physiological pH. Therefore, the lipid can be positively charged or partially positively charged at physiological pH. These lipids can be referred to as cationic or ionizable (amino) lipids. The lipids can also be zwitterionic, that is, neutral molecules with both positive and negative charges. In addition, the cationic lipid compounds described herein can have CD36 targeting, for example, any one of the lipid compounds of formula (I), (II), (III), (IV) or compounds 1-12 has a CD36 targeting higher than that of reference lipids (e.g., MC3, KC2, SM102 or DLinDMA), and the lipid nanoparticles composed thereof can be accurately recognized by the CD36 receptor and internalized into cells, releasing mRNA and expressing related proteins, or releasing other drugs. The cationic lipid compounds described herein can be advantageously used in lipid nanoparticle compositions to deliver therapeutic agents and / or prophylactic agents to mammalian cells, tissues or organs.

[0156] In one aspect, the present application provides a compound of formula (I):

[0157] Among them, R 1may be independently selected from the group consisting of hydroxy-substituted alkyl, hydroxy-substituted alkenyl, and hydroxy-substituted alkynyl;

[0158] M 1 and M 2 each independently selected from the group consisting of optionally substituted alkylene, optionally substituted alkenylene, and optionally substituted alkynylene;

[0159] L 1 and L 2 Each may be independently selected from the group consisting of -C(O)O-, -OC(O)-, -O-, -C(O)N(R)-, and -N(R)C(O)-, wherein each R is independently selected from the group consisting of: C 1-3 Alkyl, C 2-3 alkenyl and H;

[0160] M 3 and M 4 Each can independently where R 2 Can be independently selected from linear C 4-10 Alkylene, straight chain C 4-10 Alkenylene and straight chain C 4-10 The group consisting of alkynylene, R 3 Can be independently selected from linear C 4-10 Alkyl, straight chain C 4-10 Alkenyl and straight chain C 4-10 A group consisting of alkynyl groups.

[0161] In some embodiments, wherein R 2 Can be independently selected from linear C 6-9 Alkylene, straight chain C 6-9 Alkenylene and straight chain C 6-9 The group consisting of alkynylene, R 3 Can be independently selected from linear C 6-9 Alkyl, straight chain C 6-9 Alkenyl and straight chain C 6-9 For example, where R 2 Can be independently linear C 6-9 Alkylene or linear C 6-9 Alkenylene, R 3 Can be independently linear C 6-9 Alkyl or linear C 6-9 Alkenyl.

[0162] In some embodiments, wherein R 2 and R 3 They may not all be unsaturated hydrocarbon groups. 2 C 4-10 Alkylene, R 3For straight chain C 4-10 Alkenyl or straight chain C 4-10 Alkynyl. For example, R 2 For straight chain C 4-10 Alkenylene or straight chain C 4-10 Alkynylidene, R 3 For straight chain C 4-10 Alkyl. For example, R 2 and R 3 All C 4-10 Alkylene.

[0163] In some embodiments, wherein M 3 and M 4 Each can be independently a linear C 10-20 Alkenyl (e.g., C 11-19 Alkenyl, C 12-18 Alkenyl, C 13-18 Alkenyl, C 14-18 Alkenyl, C 15-18 Alkenyl or C 16-18 Alkenyl) or straight chain C 10-20 Alkynyl (e.g., C 11-19 Alkynyl, C 12-18 Alkynyl, C 13-18 Alkynyl, C 14-18 Alkynyl, C 15-18 Alkynyl or C 16-18 alkynyl).

[0164] For example, where M 3 and M 4 Each can be independently a linear C 14-18 Alkenyl (e.g. C 14 Alkenyl, C 15 Alkenyl, C 16 Alkenyl, C 17 Alkenyl or C 18 Alkenyl) or straight chain C 14-18 Alkynyl (e.g. C 14 Alkynyl, C 15 Alkynyl, C 16 Alkynyl, C 17 Alkynyl or C 18 alkynyl).

[0165] For example, where M 3 and M 4 Each can be independently a linear C 14-18 Alkenyl or straight chain C 14-18 Alkynyl, and M 3 and M 4 Each can independently where R 2 Can be independently selected from linear C 4-10Alkylene (e.g., C4 alkylene, C5 alkylene, C6 alkylene, C7 alkylene, C8 alkylene, C9 alkylene or C 10 Alkylene), R 3 Can be independently selected from linear C 4-10 Alkyl (e.g., C4 alkyl, C5 alkyl, C6 alkyl, C7 alkyl, C8 alkyl, C9 alkyl or C 10 alkyl).

[0166] In some embodiments, wherein M 3 and M 4 Each can be independently a linear C 18 Alkenyl or straight chain C 18 Alkynyl.

[0167] In some embodiments, wherein M 3 and M 4 Can be a straight chain C 18 Alkenyl.

[0168] For example, M 3 and M 4 For straight chain C 18 Alkenyl, and M 3 and M 4 Each independently where R 2 Independently selected from linear C 4-10 Alkylene (e.g., C4 alkylene, C5 alkylene, C6 alkylene, C7 alkylene, C8 alkylene, C9 alkylene or C 10 Alkylene) or linear C 4-10 Alkenylene (e.g., C4 alkenylene, C5 alkenylene, C6 alkenylene, C7 alkenylene, C8 alkenylene, C9 alkenylene or C 10 alkenylene), R 3 Independently selected from linear C 4-10 Alkyl (e.g., C4 alkyl, C5 alkyl, C6 alkyl, C7 alkyl, C8 alkyl, C9 alkyl or C 10 Alkyl) or linear C 4-10 Alkenyl (e.g., C4 alkenyl, C5 alkenyl, C6 alkenyl, C7 alkenyl, C8 alkenyl, C9 alkenyl, or C 10 alkenyl).

[0169] For example, M 3 and M 4 For straight chain C 18 Alkenyl, and M 3 and M 4 Each independently where R 2 independently selected from linear C6 alkylene, R 3Independently selected from linear C 10 Alkyl. For example, M 3 and M 4 For straight chain C 18 Alkenyl, and M 3 and M 4 Each independently where R 2 independently selected from linear C7 alkylene, R 3 is independently selected from a linear C9 alkyl group. 3 and M 4 For straight chain C 18 Alkenyl, and M 3 and M 4 Each independently where R 2 independently selected from linear C8 alkylene, R 3 is independently selected from a straight chain C8 alkyl group. 3 and M 4 For straight chain C 18 Alkenyl, and M 3 and M 4 Each independently where R 2 Independently selected from linear C 10 Alkylene, R 3 is independently selected from a straight chain C6 alkyl group. 3 and M 4 For straight chain C 18 Alkenyl, and M 3 and M 4 Each independently where R 2 independently selected from linear C9 alkylene, R 3 is independently selected from a linear C7 alkyl group. 3 and M 4 For straight chain C 18 Alkenyl, and M 3 and M 4 Each independently where R 2 independently selected from linear C8 alkylene, R 3 Independently selected from straight chain C8 alkenyl.

[0170] In some embodiments, wherein R 1 Can be independently selected from hydroxy substituted C 1-6 Alkyl, hydroxy substituted C 2-6 Alkenyl and hydroxy substituted C 2-6 For example, where R 1 C containing hydroxyl substitution1-6 Alkyl. For example, R 1 It may be selected from the group consisting of -CH2OH, -(CH2)2OH, -(CH2)3OH, -(CH2)4OH, -(CH2)5OH and -(CH2)6OH.

[0171] In some embodiments, wherein R 1 is selected from the group consisting of -CH2OH, -(CH2)2OH, -(CH2)3OH, -(CH2)4OH, -(CH2)5OH and -(CH2)6OH; and M 3 and M 4 Each independently where R 2 Independently selected from linear C 4-10 Alkylene, straight chain C 4-10 Alkenylene and straight chain C 4-10 The group consisting of alkynylene, R 3 Independently selected from linear C 4-10 Alkyl, straight chain C 4-10 Alkenyl and straight chain C 4-10 A group consisting of alkynyl groups.

[0172] In some embodiments, wherein M 1 and M 2 are each independently an optionally substituted C 1-4 Alkylene, and when M 1 and M 2 When R is C2 alkylene 1 is not -(CH2)6OH. For example, where M 1 and M 2 are each independently an optionally substituted C 3-4 Alkylene.

[0173] For example, where R 1 Selected from the group consisting of -CH2OH, -(CH2)2OH, -(CH2)3OH, -(CH2)4OH, -(CH2)5OH and -(CH2)6OH; M 1 and M 2 are each independently an optionally substituted C 3-4 Alkylene.

[0174] For example, R 1 Selected from the group consisting of -CH2OH, -(CH2)2OH, -(CH2)3OH, -(CH2)4OH, -(CH2)5OH and -(CH2)6OH; M 1 and M 2 are each independently an optionally substituted C 3-4Alkylene; M 3 and M 4 Each independently where R 2 Independently selected from linear C 4-10 Alkylene, straight chain C 4-10 Alkenylene and straight chain C 4-10 The group consisting of alkynylene, R 3 Independently selected from linear C 4-10 Alkyl, straight chain C 4-10 Alkenyl and straight chain C 4-10 A group consisting of alkynyl groups.

[0175] For example, R 1 Selected from the group consisting of -CH2OH, -(CH2)2OH, -(CH2)3OH, -(CH2)4OH, -(CH2)5OH and -(CH2)6OH; M 1 and M 2 are each independently an optionally substituted C 3-4 Alkylene; M 3 and M 4 Each independently a linear C 14-18 Alkenyl or straight chain C 14-18 Alkynyl, and M 3 and M 4 Each independently where R 2 Independently selected from linear C 4-10 Alkylene, straight chain C 4-10 Alkenylene and straight chain C 4-10 The group consisting of alkynylene, R 3 Independently selected from linear C 4-10 Alkyl, straight chain C 4-10 Alkenyl and straight chain C 4-10 A group consisting of alkynyl groups.

[0176] For example, R 1 Selected from the group consisting of -CH2OH, -(CH2)2OH, -(CH2)3OH, -(CH2)4OH, -(CH2)5OH and -(CH2)6OH; M 1 and M 2 Each independently is an optionally substituted C 3-4 Alkylene; M 3 and M 4 For straight chain C 18 Alkenyl, and M 3 and M 4 Each independently where R 2 Independently linear C 6-9Alkylene or linear C 6-9 Alkenylene, R 3 Independently linear C 6-9 Alkyl or linear C 6-9 Alkenyl.

[0177] For example, R 1 Selected from the group consisting of -CH2OH, -(CH2)2OH, -(CH2)3OH, -(CH2)4OH, -(CH2)5OH and -(CH2)6OH; M 1 and M 2 Each independently is an optionally substituted C 3-4 Alkylene; wherein M 3 and M 4 Each independently selected from:

[0178] In some embodiments, wherein the compound has formula (II):

[0179] Wherein, m is 2, 3, 4 or 5;

[0180] Each n is independently 3 or 4;

[0181] L 1 and L 2 each independently selected from the group consisting of -C(O)O-, -OC(O)-, -O-, -C(O)N(H)-, and -N(H)C(O)-;

[0182] M 3 and M 4 Each independently where R 2 Independently selected from linear C 4-10 Alkylene, straight chain C 4-10 Alkenylene and straight chain C 4-10 The group consisting of alkynylene, R 3 Independently selected from linear C 4-10 Alkyl, straight chain C 4-10 Alkenyl and straight chain C 4-10 A group consisting of alkynyl groups.

[0183] In some embodiments, wherein R 2 Independently selected from linear C 6-9 Alkylene, straight chain C 6-9 Alkenylene and straight chain C 6-9 The group consisting of alkynylene, R 3 Independently selected from linear C 6-9 Alkyl, straight chain C 6-9 Alkenyl and straight chain C 6-9 A group consisting of alkynyl groups.

[0184] In some embodiments, wherein R 2 Independently linear C 6-9 Alkylene or linear C 6-9 Alkenylene, R 3 Independently linear C 6-9 Alkyl or linear C 6-9 Alkenyl.

[0185] In some embodiments, wherein R 2 and R 3 are not unsaturated hydrocarbon groups at the same time. For example, R 2 C 6-9 Alkylene, R 3 For straight chain C 6-9 For example, R 2 For straight chain C 6-9 Alkenylene, R 3 For straight chain C 6-9 Alkyl. For example, R 2 and R 3 All C 6-9 Alkylene.

[0186] In some embodiments, wherein M 3 and M 4 Each independently a linear C 10-20 Alkenyl or straight chain C 10-20 For example, where M 3 and M 4 Each independently a linear C 14-18 Alkenyl or straight chain C 14-18 Alkynyl. For another example, where M 3 and M 4 Each independently a linear C 18 Alkenyl or straight chain C 18 Alkynyl.

[0187] In some embodiments, wherein M 3 and M 4 For straight chain C 18 For example, M 3 and M 4 For straight chain C 18 Alkenyl, and M 3 and M 4 Each independently where R 2 Independently linear C 6-9 Alkylene or linear C 6-9 Alkenylene, R 3 Independently linear C 6-9 Alkyl or linear C6-9 For example, M 3 and M 4 For straight chain C 18 Alkenyl, and M 3 and M 4 Each independently where R 2 Independently linear C 6-9 Alkylene, R 3 Independently linear C 6-9 alkyl.

[0188] In some embodiments, wherein the compound has formula (III):

[0189] Wherein, m is 2, 3 or 4;

[0190] L 1 and L 2 each independently selected from the group consisting of -C(O)O-, -OC(O)-, -CO-, -C(O)N(H)-, and -N(H)C(O)-;

[0191] M 3 and M 4 Each independently where R 2 Independently linear C 4-10 Alkylene or linear C 4-10 Alkenylene, R 3 Independently linear C 6-9 Alkyl or linear C 4-10 Alkenyl.

[0192] In some embodiments, wherein R 2 Independently linear C 6-9 Alkylene or linear C 6-9 Alkenylene, R 3 Independently linear C 6-9 Alkyl or linear C 6-9 Alkenyl.

[0193] In some embodiments, wherein R 2 and R 3 are not unsaturated hydrocarbon groups at the same time. For example, 2 and R 3 Each independently a linear C 6-9 Alkylene.

[0194] For another example, m is 2, 3 or 4; L 1 and L 2are each independently selected from the group consisting of -C(O)O-, -OC(O)-, -CO-, -C(O)N(H)-, and -N(H)C(O)-; M 3 and M 4 Each independently where R 2 and R 3 Each independently a linear C 6-9 Alkylene.

[0195] In some embodiments, wherein M 3 and M 4 For straight chain C 10-20 Alkenyl or straight chain C 10-20 For example, where M 3 and M 4 For straight chain C 14-18 Alkenyl or straight chain C 14-18 Alkynyl.

[0196] In some embodiments, wherein M 3 and M 4 For straight chain C 18 In some embodiments, wherein M 3 and M 4 For straight chain C 18 For example, M 3 and M 4 For straight chain C 18 Alkenyl, and M 3 and M 4 Each independently where R 2 Independently linear C 6-9 Alkylene or linear C 6-9 Alkenylene, R 3 Independently linear C 6-9 Alkyl or linear C 6-9 For example, M 3 and M 4 For straight chain C 18 Alkenyl, and M 3 and M 4 Each independently where R 2 Independently linear C 6-9 Alkylene, R 3 Independently linear C 6-9 alkyl.

[0197] In some embodiments, wherein M 3 and M 4 Each independently selected from:

[0198] For another example, m is 2, 3 or 4; L 1 and L 2 are each independently selected from the group consisting of -C(O)O-, -OC(O)-, -CO-, -C(O)N(H)-, and -N(H)C(O)-; M 3 and M 4 for

[0199] For example, where M 3 and M 4 for

[0200] For another example, m is 2, 3 or 4; L 1 and L 2 are each independently selected from the group consisting of -C(O)O-, -OC(O)-, -CO-, -C(O)N(H)-, and -N(H)C(O)-; M 3 and M 4 for

[0201] On the other hand, the present application provides a compound of formula (IV):

[0202] Wherein, m is 2 or 3;

[0203] L 1 and L 2 are each independently selected from the group consisting of -C(O)O-, -OC(O)-, -CO-, -C(O)N(H)- and -N(H)C(O)-.

[0204] In some embodiments, wherein L 1 and L 2 Each is independently selected from -C(O)O-, -OC(O)-, -O-, -C(O)N(H)- and -N(H)C(O)-. For example, wherein L 1 and L 2 For example, L 1 and L 2 For -OC(O)-. For another example, where L 1 and L 2 For -CO-. For another example, where L 1 and L 2 For example, L 1 and L 2 It is -N(H)C(O)-.

[0205] On the other hand, the present application provides a compound selected from:

[0206] Nanoparticle compositions

[0207] On the other hand, the application provides a nanoparticle composition, which can include a lipid component of a compound described herein and one or more additional components, such as a therapeutic and / or prophylactic agent. Nanoparticle compositions include, for example, lipid nanoparticles (LNPs), liposomes, lipid vesicles, and lipid complexes.

[0208] In some embodiments, the lipid component of the nanoparticle composition can include one or more phospholipids. The phospholipids useful in these compositions and methods can be selected from the non-limiting group consisting of: 1,2-Dilinoleoyl-sn-glycero-3-phosphocholine (DLPC), 1,2-Dimyristoyl-sn-glycero-phosphocholine (DMPC), 1,2-Dioleoyl-sn-glycero-3-phosphocholine (DOPC), 1,2-Dipalmitoyl-sn-glycero-3-phosphocholine (DPPC), 1,2-Distearoyl-sn-glycero-3-phosphocholine (DSPC), 1,2-Dicondecanoyl-sn-glycero-phosphocholine (DUPC), 1-palmitoyl-2-oleoyl-sn-glycero-3-phosphocholine (POPC), 1,2-Di-O-octadecenyl-sn-glycero-3-phosphocholine (18:0Diether PC), 1-oleoyl-2-cholesteryl hemisuccinyl-sn-glycero-3-phosphocholine (OChemsPC), 1-hexadecyl-sn-glycero-3-phosphocholine (C16 Lyso PC), 1,2-dialinolenoyl-sn-glycero-3-phosphocholine, 1,2-diarachidonoyl-sn-glycero-3-phosphocholine, 1,2-didocosahexaenoyl-sn-glycero-3-phosphocholine, 1,2-dioleoyl-sn-glycero-3-phosphoethanolamine (DOPE), 1,2-diphytanoyl-sn-glycero-3-phosphoethanolamine (ME 16.0PE), 1,2-distearoyl-sn-glycero-3-phosphoethanolamine, 1,2-dilinoleoyl-sn-glycero-3-phosphoethanolamine, 1,2-dilinolenoyl-sn-glycero-3-phosphoethanolamine, 1,2-diarachidonoyl-sn-glycero-3-phosphoethanolamine, 1,2-bisdocosahexaenoyl-sn-glycero-3-phosphoethanolamine, 1,2-dioleoyl-sn-glycero-3-phospho-rac-(1-glycerol) sodium salt (DOPG), dipalmitoylphosphatidylglycerol (DPPG), palmitoyloleoyl In one embodiment, the phospholipid may be DOPE, DOPE, DSPE, DPPE, DMPE, 1-stearoyl-2-oleoyl-stearoylethanolamine (SOPE), 1-stearoyl-2-oleoyl-phosphatidylcholine (SOPC), sphingomyelin, phosphatidylcholine, phosphatidylethanolamine, phosphatidylserine, phosphatidylinositol, phosphatidic acid, palmitoyloleoylphosphatidylcholine, lysophosphatidylcholine, lysophosphatidylethanolamine (LPE), and mixtures thereof. In one embodiment, the phospholipid may be DOPE. In another embodiment, the phospholipid is DSPC.

[0209] In some embodiments, the lipid component of the nanoparticle composition can include a structured lipid. Without limitation, the structured lipid can be selected from the group consisting of cholesterol, coprosterol, sitosterol, ergosterol, campesterol, stigmasterol, brassicasterol, tomatine, ursolic acid, α-tocopherol, and mixtures thereof. In one embodiment, the structured lipid can be cholesterol.

[0210] In some embodiments, the lipid component of the nanoparticle composition can include one or more PEG or PEG-modified lipids. These species can alternatively be referred to as PEGylated lipids. PEG lipids are lipids modified with polyethylene glycol. PEG lipids can be selected from the non-limiting group consisting of: PEG-modified phosphatidylethanolamine, PEG-modified phosphatidic acid, PEG-modified ceramide (PEG-CER), PEG-modified dialkylamines, PEG-modified diacylglycerols (PEG-DEG), PEG-modified dialkylglycerols, and mixtures thereof. For example, the PEG lipid can be selected from the group consisting of 1,2-dimyristoyl-sn-glyceromethoxypolyethylene glycol (PEG-DMG), 1,2-distearoyl-sn-glycero-3-phosphoethanolamine-N-[amino(polyethylene glycol)] (PEG-DSPE), PEG-distearylglycerol (PEG-DSG), PEG-dipalmitoleyl, PEG-dioleyl, PEG-distearyl, PEG-diacylglyceramide (PEG-DAG), PEG-dipalmitoylphosphatidylethanolamine (PEG-DPPE), or PEG-1,2-dimyristyloxypropyl-3-amine (PEG-c-DMA). For example, in some embodiments, the PEG lipid is PEG-DMG.

[0211] In some embodiments, the PEG lipid comprises a PEG moiety having a size of about 1000 Da to about 20 kDa.

[0212] In some embodiments, the PEG lipid is selected from any one or a combination of more than one of PEG1000-DMG, PEG5000-DMG, PEG2000-DMG and PEG2000-DSPE.

[0213] In some embodiments, the PEG lipid is PEG2000-DMG.

[0214] In some embodiments, the lipid component further comprises phospholipids, structural lipids and PEG lipids.

[0215] The lipid component of the nanoparticle composition can include, for example, a compound according to Formula (I), (II), (III), (IV) or compounds 1-12, a phospholipid (such as an unsaturated lipid, such as DOPE or DSPC), a PEG lipid, and a structural lipid. The ingredients of the provided lipid component can be in a specific ratio.

[0216] In some embodiments, the lipid component comprises about 30 mol% to about 60 mol% of the compound, about 0 mol% to about 30 mol% of phospholipids, about 18.5 mol% to about 48.5 mol% of structural lipids, and about 0 mol% to about 10 mol% of PEG lipids. In some embodiments, the lipid component of the nanoparticle composition comprises about 35 mol% to about 55 mol% of the compound of formula (I), (II), (III), or (IV); about 5 mol% to about 25 mol% of phospholipids; about 30 mol% to about 40 mol% of structural lipids; and about 0 mol% to about 10 mol% of PEG lipids. In a specific embodiment, the lipid component comprises about 50 mol% of the compound, about 10 mol% of phospholipids, about 38.5 mol% of structural lipids, and about 1.5 mol% of PEG lipids. In another specific embodiment, the lipid component comprises about 40 mol% of the compound, about 20 mol% of phospholipids, about 38.5 mol% of structural lipids, and about 1.5 mol% of PEG lipids. In some embodiments, the phospholipid can be DOPE or DSPC. In other embodiments, the PEG lipid can be PEG-DMG and / or the structural lipid can be cholesterol.

[0217] For example, the lipid component of the nanoparticle composition can include about 50 mol% of Compound 1 or Compound 2, about 10 mol% of DOPE or DSPC, about 37.0-39.5 mol% of cholesterol or sitosterol, and about 0.5-3.0 mol% of PEG2000-DMG.

[0218] For another example, the lipid component of the nanoparticle composition can include about 50 mol% of Compound 1 or Compound 2, about 10 mol% of DOPE or DSPC, about 38.5 mol% of cholesterol or sitosterol, and about 1.5 mol% of PEG2000-DMG.

[0219] Nanoparticle compositions can include one or more therapeutic and / or prophylactic agents. Therapeutic and / or prophylactic agents include biologically active substances and are alternatively referred to as "active agents." Therapeutic and / or prophylactic agents can be substances that, after delivery to a cell or organ, cause desired changes in the cell or organ or in other body tissues or systems. Such species can be used to treat one or more diseases, disorders, or conditions. In some embodiments, the therapeutic and / or prophylactic agent is an anticancer agent, an antiviral agent, an immunomodulator, an anti-inflammatory agent, or an agent that regulates cellular metabolic activity.

[0220] In some embodiments, the therapeutic and / or prophylactic agent is a nucleic acid, a protein, a peptide, or a small molecule.

[0221] In some embodiments, the therapeutic and / or preventive agent is a nucleic acid. Exemplary nucleic acids include, but are not limited to, one or more of the following: deoxyribonucleic acid (DNA); ribonucleic acid (RNA), including messenger mRNA (mRNA), its hybrid; RNAi inducing factor; RNAi factor; siRNA; shRNA; miRNA; antisense RNA; ribozyme; catalytic DNA; RNA that induces triple helix formation; aptamer; carrier, etc. In some embodiments, the therapeutic and / or preventive agent is RNA. The RNA that can be used in the compositions and methods described herein can be selected from, but is not limited to, the group consisting of: shortmer, antagomir, antisense RNA, ribozyme, small interfering RNA (siRNA), asymmetric interfering RNA (aiRNA), microRNA (miRNA), Dicer-substrate RNA (dsRNA), small hairpin RNA (shRNA), transfer RNA (tRNA), messenger RNA (mRNA), and mixtures thereof.

[0222] In some embodiments, the therapeutic and / or preventive agent can be mRNA. The mRNA can encode any polypeptide of interest, including any naturally occurring or non-naturally occurring or otherwise modified polypeptide. The polypeptide encoded by the mRNA can be of any size and can have any secondary structure or activity. In some embodiments, the polypeptide encoded by the mRNA can have a therapeutic effect when expressed in a cell. For example, the mRNA can encode preventive or therapeutic proteins such as green fluorescent protein, myoglobin, myosin, fetal hemoglobin, collagen, tolerance-inducing autoantigen proteins, and the novel coronavirus S protein.

[0223] In other embodiments, therapeutic agent and / or preventive agent can be siRNA.siRNA can selectively reduce the expression of paid close attention to gene or lower the expression of this gene.For example, the selection of siRNA can make after the nanoparticle composition that will comprise this siRNA is administered to experimenter in need, make the gene silencing relevant with specific disease, disease or the patient's condition.siRNA can comprise the sequence complementary to the mRNA sequence of paid close attention to gene or protein with coding.In some embodiments, siRNA can be immunomodulatory siRNA.

[0224] In other embodiments, therapeutic agent and / or preventive agent can be shRNA or its encoding vector or plasmid.shRNA can produce inside target cell after suitable construct is delivered in the core.Construct and mechanism relevant to shRNA are well-known in the related art.

[0225] In some embodiments, the therapeutic agent and / or preventive agent comprises a component of a gene editing system. The term "gene editing" and its grammatical equivalents may refer to a genetic engineering that inserts, replaces or removes one or more nucleotides from a genome. For example, gene editing can be performed using a nuclease that includes a CRISPR-associated protein (Cas protein, such as Cas9), a zinc finger nuclease (ZFN), a transcription activator-like effector nuclease (TALEN) and a large range of nucleases. The nuclease may be a naturally occurring nuclease, a genetically modified nuclease and / or a recombinant nuclease. For example, the CRISPR / cas system may be suitable as a gene editing system. The term "gene editing system" refers to a system, such as one or more molecules, that guides and achieves changes (e.g., deletions) of one or more nucleic acids at or near the targeted genomic DNA site by the system.

[0226] In some embodiments, the components of the gene editing system include polynucleotides encoding nucleases (e.g., zinc finger nucleases (ZFNs), transcription activator-like effector nucleases (TALENs), CRISPR-associated protein 9 (Cas9), and engineered homing endonucleases).

[0227] In some embodiments, wherein the nuclease is Cas9 and the nanoparticle composition further comprises a guide RNA that targets the nuclease to a specific site in the genome of the target cell. In some embodiments, the guide RNA can comprise two RNA molecules, referred to herein as "dual guide RNA" or "dgRNA." In other embodiments, the guide RNA can comprise a single RNA molecule, referred to herein as "single guide RNA" or "sgRNA."

[0228] In some embodiments, the components of the gene editing system include: i) CRISPR-associated protein 9 (Cas9), or mRNA encoding CRISPR-associated protein 9 (Cas9); and ii) single guide RNA (sgRNA), or a nucleic acid encoding a single guide RNA (sgRNA).

[0229] For example, the therapeutic and / or prophylactic agent can be sgRNA and / or cas9 mRNA. sgRNA and / or cas9 mRNA can be used as a gene editing tool. For example, the sgRNA-cas9 complex can affect the mRNA translation of a cellular gene.

[0230] In other embodiments, the therapeutic and / or prophylactic agent can be a protein. The therapeutic proteins that can be used in the nanoparticles in the present application include, but are not limited to, gentamicin, amikacin, insulin, erythropoietin (EPO), granulocyte colony stimulating factor (G-CSF), granulocyte-macrophage colony stimulating factor (GM-CSF), factor VIR, luteinizing hormone-releasing hormone (LHRH) analogs, interferon, heparin, hepatitis B surface antigen, typhoid vaccine, and cholera vaccine.

[0231] In other embodiments, the therapeutic and / or prophylactic agent comprises a small molecule. For example, the therapeutic and / or prophylactic agent can be a cytotoxin, a radioactive ion, a chemotherapeutic agent, a vaccine, or a compound that elicits an immune response.

[0232] In some embodiments, the therapeutic and / or prophylactic agent may include a PROTAC (Proteolysis Targeting Chimera) molecule, also known as a proteolysis targeting chimeric molecule. PROTAC is a heterobifunctional molecule that is generally designed to have three parts: 1) a ligand / molecule that binds to and / or regulates a ubiquitin ligase; 2) a small molecule that binds to the target protein of interest for proteolysis; and 3) a linker that connects the two molecules together. Therefore, PROTAC works by allowing the ligand / molecule to bind to the ubiquitin ligase, thereby recruiting the target of the protein of interest to the ligase for ubiquitination and ultimately proteolysis and degradation. Without limitation, the PROTAC may include ARV-110, ARV-471, ARV-766, ARV-771, AVR-825, AR-LDD, DT2216, KT-474, KT-413, KT-333, NX-2127, NX-5948, CG001419, CFT8634, FHD-609 or SARD279.

[0233] In the nanoparticle composition, the amount of therapeutic and / or prophylactic agent can depend on the size, composition, desired target and / or application or other characteristics of the nanoparticle composition, and the characteristic of the therapeutic and / or prophylactic agent. For example, the amount of the RNA that can be used for the nanoparticle composition can depend on the size, sequence and other characteristics of the RNA. The relative amount of therapeutic and / or prophylactic agent and other compositions (lipid) can also change in the nanoparticle composition. In some embodiments, the wt / wt ratio of lipid component and therapeutic and / or prophylactic agent can be about 5:1 to about 60:1, such as 5:1, 6:1, 7:1, 8:1, 9:1, 10:1, 11:1, 12:1, 13:1, 14:1, 15:1, 16:1, 17:1, 18:1, 19:1, 20:1, 25:1, 30:1, 35:1, 40:1, 45:1, 50:1 and 60:1. For example, the wt / wt ratio of the lipid component to the therapeutic and / or prophylactic agent can be about 10: 1 to about 40: 1. In certain embodiments, the wt / wt ratio is about 20: 1. The amount of the therapeutic and / or prophylactic agent in the nanoparticle composition can be measured, for example, using absorption spectroscopy (e.g., UV-visible spectroscopy).

[0234] In some embodiments, the nanoparticle combination includes one or more RNAs, and the one or more RNAs, lipids, and amounts thereof selected can provide a specific N:P ratio. The N:P ratio of the composition refers to the ratio of the molar ratio of nitrogen atoms in the one or more lipids to the number of phosphate groups in the RNA. In general, a lower N:P ratio is preferred. The one or more RNAs, lipids, and amounts thereof selected can provide an N:P ratio of about 2:1 to about 30:1, such as 2:1, 3:1, 4:1, 5:1, 6:1, 7:1, 8:1, 9:1, 10:1, 12:1, 14:1, 16:1, 18:1, 20:1, 22:1, 24:1, 26:1, 28:1, or 30:1. In certain embodiments, the N:P ratio can be about 2:1 to about 8:1. In other embodiments, the N:P ratio is about 5:1 to about 8:1. For example, the N:P ratio can be about 5.0: 1, about 5.5: 1, about 5.67: 1, about 6.0: 1, about 6.5: 1, or about 7.0: 1. For example, the N:P ratio can be about 6:1.

[0235] Nanoparticle compositions can be characterized by a variety of methods. For example, microscopy (e.g., transmission electron microscopy or scanning electron microscopy) can be used to examine the morphology and size distribution of the nanoparticle compositions. Dynamic light scattering or potentiometric analysis (e.g., potentiometric titration) can be used to measure zeta potential. Dynamic light scattering can also be used to measure particle size. Multiple characteristics of the nanoparticle compositions, such as particle size, polydispersity index, and zeta potential, can also be measured using instruments such as the Zetasizer Nano ZS (Malvern Instruments Ltd, Malvern, Worcestershire, UK).

[0236] The nanoparticle compositions can have an average size, for example, between tens of nanometers and hundreds of nanometers, as measured by dynamic light scattering (DLS). For example, the average size can be from about 40 nm to about 150 nm, such as about 40 nm, 45 nm, 50 nm, 55 nm, 60 nm, 65 nm, 70 nm, 75 nm, 80 nm, 85 nm, 90 nm, 95 nm, 100 nm, 105 nm, 110 nm, 115 nm, 120 nm, 125 nm, 130 nm, 135 nm, 140 nm, 145 nm, or 150 nm. In some embodiments, the average size of the nanoparticle composition can be from about 50 nm to about 100 nm, from about 50 nm to about 90 nm, from about 50 nm to about 80 nm, from about 50 nm to about 70 nm, from about 50 nm to about 60 nm, from about 60 nm to about 100 nm, from about 60 nm to about 90 nm, from about 60 nm to about 80 nm, from about 60 nm to about 70 nm, from about 70 nm to about 150 nm, from about 70 nm to about 130 nm, from about 70 nm to about 100 nm, from about 70 nm to about 90 nm, from about 70 nm to about 80 nm, from about 80 nm to about 150 nm, from about 80 nm to about 130 nm, from about 80 nm to about 100 nm, from about 80 nm to about 90 nm, from about 90 nm to about 150 nm, from about 90 nm to about 130 nm, or from about 90 nm to about 100 nm. In certain embodiments, the average size of the nanoparticle composition can be from about 70 nm to about 130 nm or from about 70 nm to about 100 nm. In one particular embodiment, the average size can be about 80 nm. In other embodiments, the average size can be about 100 nm. In other embodiments, the average size can be about 120 nm.

[0237] In some embodiments, the polydispersity index of a nanoparticle composition can be about 0 to about 0.25, such as 0.01, 0.02, 0.03, 0.04, 0.05, 0.06, 0.07, 0.08, 0.09, 0.10, 0.11, 0.12, 0.13, 0.14, 0.15, 0.16, 0.17, 0.18, 0.19, 0.20, 0.21, 0.22, 0.23, 0.24 or 0.25. In some embodiments, the polydispersity index of a nanoparticle composition can be about 0.10 to about 0.20.

[0238] The zeta potential of a nanoparticle composition can be used to indicate the zeta potential of the composition. For example, the zeta potential can describe the surface charge of a nanoparticle composition. Nanoparticle compositions with relatively low charge, i.e., positively or negatively charged, are generally desirable because species with higher charges may interact undesirably with cells, tissues, and other elements in the body. In some embodiments, the zeta potential of the nanoparticle composition can be from about -10 mV to about +20 mV, about -10 mV to about +15 mV, about -10 mV to about +10 mV, about -10 mV to about +5 mV, about -10 mV to about 0 mV, about -10 mV to about -5 mV, about -5 mV to about +20 mV, about -5 mV to about +15 mV, about -5 mV to about +10 mV, about -5 mV to about +5 mV, about -5 mV to about 0 mV, about 0 mV to about +20 mV, about 0 mV to about +15 mV, about 0 mV to about +10 mV, about 0 mV to about +5 mV, about +5 mV to about +20 mV, about +5 mV to about +15 mV, or about +5 mV to about +10 mV.

[0239] Pharmaceutical composition

[0240] Nanoparticle compositions can be formulated as whole or as part of a pharmaceutical composition. A pharmaceutical composition can include one or more nanoparticle compositions. For example, a pharmaceutical composition can include one or more nanoparticle compositions containing one or more different therapeutic and / or prophylactic agents. Pharmaceutical compositions can also include one or more pharmaceutically acceptable carriers, such as, but not limited to, one or more solvents, dispersion media, diluents, dispersing aids, suspending aids, granulation aids, disintegrants, fillers, glidants, liquid vehicles, binders, surfactants, isotonic agents, thickeners or emulsifiers, buffers, lubricants, oils, preservatives, and other species. Excipients such as waxes, butters, colorants, coatings, flavorings, and fragrances can also be included.

[0241] Nanoparticle compositions and / or pharmaceutical compositions comprising one or more nanoparticle compositions can be administered to any patient or subject, including patients or subjects who can benefit from the therapeutic effect provided by delivering therapeutic and / or prophylactic agents to one or more specific cells, tissues, organs or systems or a combination thereof. Although the description of nanoparticle compositions and pharmaceutical compositions comprising nanoparticle compositions provided herein is primarily directed to compositions suitable for administration to humans, it will be appreciated by those skilled in the art that these compositions are generally suitable for administration to any other mammal. It will be fully appreciated that compositions suitable for administration to humans can be modified so that these compositions are suitable for administration to various animals, and that such modifications can be designed and / or performed by ordinary skilled veterinary pharmacologists with only routine experimentation (if any) required. It is contemplated that subjects to whom these compositions may be administered include, but are not limited to, humans, other primates, and other mammals, including commercially relevant mammals such as cattle, pigs, horses, sheep, cats, dogs, mice, and / or rats.

[0242] Pharmaceutical compositions can be prepared into various forms suitable for a variety of routes and methods of administration. For example, pharmaceutical compositions can be prepared into liquid dosage forms (e.g., emulsions, microemulsions, nanoemulsions, solutions, suspensions, syrups, and elixirs), injectable forms, solid dosage forms (e.g., capsules, tablets, pills, powders, and granules), dosage forms for topical and / or transdermal administration (e.g., ointments, pastes, creams, lotions, gels, powders, solutions, sprays, inhalants, and patches), suspensions, powders, and other forms. Pharmaceutical compositions comprising one or more nanoparticle compositions can be prepared by any method known or later developed in the art of pharmacology.

[0243] Delivery system

[0244] CD36 is a transmembrane protein of the class B scavenger receptor family, also known as FAT, SCARB3, GP88, glycoprotein IV (gpIV), and glycoprotein IIIb (gpIIIb). This protein is widely expressed on many cells, such as microvascular endothelial cells, macrophages, platelets, adipocytes, epithelial cells (e.g., intestinal epithelial cells and renal tubular cells), pancreatic islet cells, and cardiomyocytes. CD36 recognizes and binds to a variety of ligands, including oxidized or modified low-density lipoproteins (oxLDL, mLDL); long-chain fatty acids (LCFA); lipid and lipoprotein components of bacterial cell walls; thrombospondins (TSP)-1 and -2 and molecules with type 1 thrombospondin repeat (TSR) peptide domains; fibrillar β-amyloid (fAβ); dying / apoptotic cells; glycated proteins; and phospholipids. Ligand recognition by CD36 initiates a signaling cascade in phagocytes (such as macrophages) resulting in the engulfment of the ligand (such as lipids and fatty acids) and any other material linked to the ligand, such as cellular components, bacteria, etc.

[0245] CD36 has been shown to bind to nanoparticle compositions containing a compound according to Formula (I), (II), (III), (IV), or Compounds 1-12 in vitro and in vivo, and is known to bind to the CD36 receptor found on the surface of cells. Therefore, administering a nanoparticle composition containing a compound according to Formula (I), (II), (III), (IV), or Compounds 1-12 to a subject can bind to CD36 in vivo in the subject and can subsequently deliver therapeutic and / or prophylactic agents (e.g., RNA) to cells expressing CD36 in a targeted manner.

[0246] Nanoparticle compositions can be designed for one or more specific applications or targets. For example, nanoparticle compositions can be designed to deliver therapeutic and / or prophylactic agents, such as RNA, to specific cells, tissues, organs, or systems, or combinations thereof, in a mammal (e.g., cells and tissues expressing CD36).

[0247] The application provides the method that therapeutic agent and / or preventative are delivered to mammalian cell or organ.Therapeutic agent and / or preventative are delivered to cell and relate to and will comprise the nanoparticle composition of this therapeutic agent and / or preventative and be administered to experimenter, wherein the using of said composition relates to and makes this cell contact with said composition.For example, protein, cytotoxic agent, radioactive ion, chemotherapeutic agent or nucleic acid (as RNA, for example mRNA) can utilize nanoparticle composition to be delivered to cell or organ.When therapeutic agent and / or preventative are mRNA, after cell contacts with nanoparticle composition, translatable mRNA can produce paid close attention to polypeptide in cell translation.

[0248] The nanoparticle compositions of the present application can target specific types or categories of cells (e.g., cells of a specific organ or its system). For example, a nanoparticle composition comprising a therapeutic and / or prophylactic agent of interest can be specifically delivered to mammalian muscle, liver, kidney, spleen, femur, or lung. Specific delivery to a specific category of cells, organs, or systems, or a combination thereof, indicates that, for example, after the nanoparticle composition is administered to a mammal, a higher proportion of the nanoparticle composition comprising the therapeutic and / or prophylactic agent is delivered to the destination of interest (e.g., tissue expressing CD36) relative to other targets. In some embodiments, specific delivery can increase the amount of the therapeutic and / or prophylactic agent per 1 g of tissue of the target destination (e.g., tissue of interest, such as tissue expressing CD36) by more than 2 times, 5 times, 10 times, 15 times, or 20 times compared to another destination. In some embodiments, the tissue of interest is selected from the group consisting of muscle (e.g., by intramuscular injection), liver, kidney, lung, spleen, femur, eye tissue (e.g., by intraocular, subretinal, or intravitreal injection), vascular endothelium in a blood vessel (e.g., within the coronary or femoral artery) or kidney, and tumor tissue (e.g., by intratumoral injection).

[0249] The present application provides uses of the compounds and nanoparticle compositions described herein in preparing delivery systems.

[0250] In another aspect, the present application provides a delivery system comprising the compound described herein or the nanoparticle composition described herein.

[0251] In some embodiments, the delivery system is a CD36-targeted delivery system. For example, the delivery system of the present application can "enhance delivery" and / or "specifically deliver" therapeutic agents and / or prophylactic agents to cells, tissues, or organs that express CD36.

[0252] In another aspect, the present application provides a method for delivering a therapeutic and / or prophylactic agent to mammalian cells, tissues and / or organs, the method comprising administering the nanoparticle composition described herein to a subject in need thereof, wherein the administration comprises contacting the cell with the nanoparticle composition, thereby delivering the therapeutic and / or prophylactic agent to the cell.

[0253] In some embodiments, the cells, tissues and / or organs express CD36.

[0254] In some embodiments, the cells, tissues and / or organs highly express CD36.

[0255] In some embodiments, the mammalian cell is in vivo or in vitro in a mammal.

[0256] In some embodiments, the mammal is a human.

[0257] Methods and uses

[0258] In another aspect, the present application provides a method for preventing and / or treating a disease or condition, comprising administering an effective amount of the nanoparticle composition described herein to a subject in need thereof.

[0259] The present application also provides use of the nanoparticle composition described herein in preparing medicines.

[0260] The present application also provides a drug for preventing and / or treating a disease or condition, which comprises the nanoparticle composition described in the present application.

[0261] The methods provided herein relate to administering nanoparticle compositions containing one or more therapeutic and / or prophylactic agents and pharmaceutical compositions comprising these nanoparticle compositions. For the features and embodiments of the present application, the terms therapeutic agent and prophylactic agent can be used interchangeably herein. Therapeutic compositions or imaging, diagnostic or prophylactic compositions can be administered to a subject using any reasonable amount and any route of administration that is effective for preventing, treating, diagnosing or imaging a disease, disorder and / or condition and / or for any other purpose. The specific amount administered to a given subject can vary depending on the species, age and general condition of the subject; the purpose of administration; the specific composition; the mode of administration, etc. The compositions according to the present application can be formulated into unit dosage forms to facilitate administration and uniformity of dosage. However, it should be understood that the total daily dosage of the compositions of the present application will be determined by the attending physician within the scope of reasonable medical judgment. The specific therapeutically effective, prophylactically effective, or other appropriate dosage level (e.g., for imaging) for any particular patient will depend on a variety of factors, including the severity and nature of the condition being treated, if any; the therapeutic and / or prophylactic agent(s) employed; the specific composition employed; the patient's age, weight, general health, sex, and diet; the time of administration, route of administration, and rate of excretion of the specific pharmaceutical composition employed; the duration of the treatment; drugs used in combination or concomitantly with the specific pharmaceutical composition employed; and like factors well known in the medical arts.

[0262] Nanoparticle compositions containing one or more therapeutic and / or prophylactic agents can be administered by any route. In some embodiments, compositions comprising one or more nanoparticle compositions described herein, including prophylactic, diagnostic, or imaging compositions, are administered by one or more of a variety of routes, including oral, intravenous, intramuscular, intraarterial, intramedullary, intrathecal, intraparenchymal, subcutaneous, intraventricular, transdermal or intradermal, interdermal, rectal, intravaginal, intraperitoneal, intraocular, subretinal, intravitreal, topical (e.g., by powder ointment, cream, gel, lotion, and / or drops), mucosal, nasal, buccal, intestinal, vitreous, intratumoral, sublingual, intranasal; by intratracheal instillation, bronchial instillation, and / or inhalation; in the form of an oral spray and / or powder, nasal spray, and / or aerosol, and / or through a portal vein catheter. In some embodiments, the composition can be administered intravenously, intramuscularly, intradermally, intraarterially, intratumorally, subcutaneously, intraocular, subretinal, intravitreal, intraparenchymal, or by any other parenteral route of administration or by inhalation. However, taking into account possible advances in drug delivery science, the present disclosure encompasses delivery or administration of the compositions described herein by any appropriate route. In general, the most appropriate route of administration will depend on a variety of factors, including the properties of the nanoparticle composition containing one or more therapeutic and / or prophylactic agents (e.g., its stability in various body environments such as the bloodstream and the gastrointestinal tract), the patient's condition (e.g., whether the patient can tolerate a particular route of administration), etc.

[0263] Nanoparticle compositions comprising one or more therapeutic and / or prophylactic agents can be used in combination with one or more other therapeutic, prophylactic, diagnostic, or imaging agents. "In combination with" is not intended to indicate that these agents must be administered at the same time and / or formulated for delivery together, but these delivery methods are within the scope of this disclosure. For example, one or more nanoparticle compositions comprising one or more different therapeutic and / or prophylactic agents can be administered in combination. The composition can be administered simultaneously with, before, or after one or more other desired therapeutic agents or medical procedures. Generally speaking, each agent will be administered at a dose and / or time course determined for that agent. In some embodiments, this disclosure encompasses a combination of a delivery composition or its imaging, diagnostic, or prophylactic composition with an agent that improves its bioavailability, reduces and / or improves its metabolism, inhibits its excretion, and / or improves its distribution in the body. In addition, it should be understood that the therapeutic, prophylactic, diagnostic, or imaging active agents used in combination can be administered together in a single composition or separately in different compositions. Generally speaking, it is expected that the amount of the agents used in combination will not exceed the level at which they would be used independently. In some embodiments, the level used in combination may be lower than the level used independently.

[0264] Example

[0265] The following will clearly and completely describe the technical solutions of the present invention in conjunction with the accompanying drawings. Obviously, the embodiments described are only some of the embodiments of the present invention, not all of them. All other embodiments obtained by ordinary technicians in this field based on the embodiments of the present invention without making any creative efforts shall fall within the scope of protection of the present invention.

[0266] Example 1 Synthesis of di((Z)-octadec-9-en-1-yl)4,4'-((2-hydroxyethyl)azanediyl)dibutyrate

[0267] Step 1: Synthesis of (Z)-octadec-9-en-1-yl 4-bromobutanoate

[0268] 4-Butylbromoacid (5 g, 29.94 mmol) and oleyl alcohol (8.04 g, 29.94 mmol) were mixed and heated to 65°C. After the reaction mixture was completely melted, 3 drops of concentrated sulfuric acid were added and stirring continued at 65°C overnight. After the reaction was completed, heating was stopped and 100 mL of EA and 100 mL of a saturated NaHCO3 solution were added for washing. The aqueous phase was removed and the organic phase was washed with saturated brine, dried over anhydrous sodium sulfate, and spin-dried through a column. Column chromatography was performed using PE:EA = 10:1 as the eluent to give 9.89 g of the product as a colorless oil in a 76% yield.

[0269] Step 2: Synthesis of di((Z)-octadec-9-en-1-yl)4,4'-((2-hydroxyethyl)azanediyl)dibutyrate

[0270] Ethanolamine (500 mg, 8.19 mmol) was dissolved in 50 mL of MeCN. (Z)-octadec-9-en-1-yl4-bromobutanoate (8.54 g, 20.46 mmol), K2CO3 (7.02 g, 50.770 mmol), and KI (20 mg) were added and stirred at 85°C overnight. 100 mL of water and 100 mL of EA were added. After separation, the organic phase was washed with saturated brine, dried over anhydrous sodium sulfate, and then dried by column chromatography. Column chromatography was performed using DCM (containing 5% EA):MeOH = 50:1 to 5:1 to obtain 2.35 g of the product as a light yellow oil, yield: 39.1%. m / z [m+H + ]:734.66

[0271] Example 2 Synthesis of ((2-hydroxyethyl)azanediyl)bis(propane-3,1-diyl)(10Z,10'Z)-bis(octadec-10-enoate)

[0272] Step 1: Synthesis of 3-bromopropyl oleate

[0273] 1-Bromobutanol (5.42g, 35.4mmol) and oleic acid (10g, 35.4mmol) were mixed and heated to 65°C. After the reaction mixture was completely melted, 3 drops of concentrated sulfuric acid were added and stirring continued at 65°C overnight. After the reaction was completed, heating was stopped and 100mL of EA and 100mL of NaHCO3 saturated solution were added for washing. The aqueous phase was removed and the organic phase was washed with saturated salt water and then dried over anhydrous sodium sulfate and spin-dried for column. Column chromatography was eluted with PE:EA=10:1 to give 10.6g of the product as a colorless oil with a yield of 71%.

[0274] Step 2: ((2-hydroxyethyl)azanediyl)bis(propane-3,1-diyl)(10Z,10'Z)-bis(octadec-10-enoate) Synthesis

[0275] Ethanolamine (500 mg, 8.19 mmol) was dissolved in 50 mL of MeCN. (Z)-octadec-9-en-1-yl4-bromobutanoate (8.26 g, 20.46 mmol), K2CO3 (7.02 g, 50.770 mmol), and KI (20 mg) were added and stirred at 85°C overnight. 100 mL of water and 100 mL of EA were added. After separation, the organic phase was washed with saturated brine, dried over anhydrous sodium sulfate, and then dried by column chromatography. Column chromatography was performed using DCM (containing 5% EA):MeOH = 50:1 to 5:1 to obtain 3.95 g of the product as a light yellow oil in a yield of 68.3%. m / z [m+H + ]:734.66

[0276] Example 3 Synthesis of 4,4'-((2-hydroxyethyl)azanediyl)bis(N-((Z)-octadec-9-en-1-yl)butanamide)

[0277] Step 1: Synthesis of (Z)-4-bromo-N-(octadec-9-en-1-yl)butanamide

[0278] Under nitrogen, 4-butylbromoic acid (5 g, 29.94 mmol) and (Z)-octadec-9-en-1-amine (8.81 g, 32.93 mmol) were mixed, and 100 mL of DCM and a small amount of molecular sieves were added. After the reaction mixture was completely dissolved, 0.1 eq of DMAP and 2 eq of DCC were added and stirred overnight. After the reaction was complete, 100 mL of EA and 100 mL of saturated NaHCO3 solution were added for washing. The aqueous phase was removed, and the organic phase was washed with saturated brine, dried over anhydrous sodium sulfate, and passed through a column. Column chromatography with PE:EA = 10:1 was used as the eluent to obtain 6.89 g of the product as a colorless oil, with a yield of 55%.

[0279] Step 2: Synthesis of 4,4'-((2-hydroxyethyl)azanediyl)bis(N-((Z)-octadec-9-en-1-yl)butanamide

[0280] 2-aminoethan-1-ol (100 mg, 1.64 mmol) was dissolved in 50 mL of MeCN. (Z)-4-bromo-N-(octadec-9-en-1-yl)butanamide (2.05 g, 4.91 mmol), K2CO3 (3 g, 21.7 mmol), and KI (20 mg) were added and stirred at 85°C overnight. 100 mL of water and 100 mL of EA were added. After separation, the organic phase was washed with saturated brine, dried over anhydrous sodium sulfate, and then dried by column chromatography. Column chromatography was performed using DCM (containing 5% EA):MeOH (containing 10% NH3) in a ratio of 50:1 to 5:1 to afford 0.72 g of the product as a light yellow oil in a 60% yield. m / z [m+H + ]:733.24

[0281] Example 4 Synthesis of 2-(bis(4-(((Z)-octadec-9-en-1-yl)oxy)butyl)amino)ethan-1-ol

[0282] Step 1: Synthesis of (Z)-1-(4-bromobutoxy)octadec-9-ene

[0283] Combine 4-bromobutan-1-ol (5 g, 32.68 mmol) and (Z)-octadec-9-en-1-ol (9.65 g, 35.94 mmol) and heat to 120°C. Once the reaction mixture is completely melted, add 3 drops of concentrated sulfuric acid and continue stirring at 120°C overnight. After the reaction is complete, remove the heat and wash with 100 mL of EA and 100 mL of saturated NaHCO₃ solution. Remove the aqueous phase, wash the organic phase with saturated brine, dry it over anhydrous sodium sulfate, and filter it through a column. Column chromatography using PE:EA = 10:1 yields 9.89 g of the product as a colorless oil (75% yield).

[0284] Step 2: Synthesis of 2-(bis(4-(((Z)-octadec-9-en-1-yl)oxy)butyl)amino)ethan-1-ol

[0285] 2-aminoethan-1-ol (200 mg, 3.27 mmol) was dissolved in 50 mL of MeCN. (Z)-1-(4-bromobutoxy)octadec-9-ene (3.96 g, 9.82 mmol), K2CO3 (5 g, 36.2 mmol), and KI (20 mg) were added and stirred at 85°C overnight. 100 mL of water and 100 mL of EA were added. After separation, the organic phase was washed with saturated brine, dried over anhydrous sodium sulfate, and then dried by column chromatography. Column chromatography was performed using DCM (containing 5% EA):MeOH (containing 10% NH3) in a ratio of 50:1 to 5:1 to afford 1.02 g of the product as a light yellow oil in a yield of 44.1%. m / z [m+H + ]706.7.

[0286] Example 5 Synthesis of di((Z)-octadec-9-en-1-yl)4,4'-((3-hydroxypropyl)azanediyl)dibutyrate

[0287] Step 1: Synthesis of (Z)-octadec-9-en-1-yl 4-bromobutanoate

[0288] 4-Butylbromoacid (5 g, 29.94 mmol) and oleyl alcohol (8.04 g, 29.94 mmol) were mixed and heated to 65°C. After the reaction mixture was completely melted, 3 drops of concentrated sulfuric acid were added and stirring continued at 65°C overnight. After the reaction was completed, heating was stopped and 100 mL of EA and 100 mL of a saturated NaHCO3 solution were added for washing. The aqueous phase was removed and the organic phase was washed with saturated brine, dried over anhydrous sodium sulfate, and spin-dried through a column. Column chromatography was performed using PE:EA = 10:1 as the eluent to give 9.89 g of the product as a colorless oil in a 76% yield.

[0289] Step 2: Synthesis of di((Z)-octadec-9-en-1-yl)4,4'-((3-hydroxypropyl)azanediyl)dibutyrate

[0290] Propanolamine (500 mg, 6.66 mmol) was dissolved in 50 mL of MeCN. (Z)-octadec-9-en-1-yl 4-bromobutanoate (6.95 g, 16.64 mmol), K2CO3 (7.02 g, 50.770 mmol), and KI (20 mg) were added and stirred at 85°C overnight. 100 mL of water and 100 mL of EA were added. After separation, the organic phase was washed with saturated brine, dried over anhydrous sodium sulfate, and then dried by column chromatography. Column chromatography was performed using DCM (containing 5% EA):MeOH = 50:1 to 5:1 to obtain 3.1 g of the product as a light yellow oil in a yield of 62.2%. m / z [m+H + ]:749.23

[0291] Example 6 Synthesis of 4,4'-((3-hydroxypropyl)azanediyl)bis(N-((Z)-octadec-9-en-1-yl)butanamide)

[0292] Step 1: Synthesis of (Z)-4-bromo-N-(octadec-9-en-1-yl)butanamide

[0293] Under nitrogen, 4-butylbromoic acid (5 g, 29.94 mmol) and (Z)-octadec-9-en-1-amine (8.81 g, 32.93 mmol) were mixed, and 100 mL of DCM and a small amount of molecular sieves were added. After the reaction mixture was completely dissolved, 0.1 eq of DMAP and 2 eq of DCC were added and stirred overnight. After the reaction was completed, 100 mL of EA and 100 mL of saturated NaHCO3 solution were added for washing. The aqueous phase was removed, and the organic phase was washed with saturated brine, dried over anhydrous sodium sulfate, and dried over a column. Column chromatography was performed using PE:EA = 10:1 as the eluent to obtain 6.89 g of the product as a colorless oil with a yield of 55%.

[0294] Step 2: Synthesis of 4,4'-((3-hydroxypropyl)azanediyl)bis(N-((Z)-octadec-9-en-1-yl)butanamide)

[0295] Propanolamine (400 mg, 5.33 mmol) was dissolved in 50 mL of MeCN, and (Z)-octadec-9-en-1-yl4-bromobutanoate (5.54 g, 13.31 mmol), K2CO3 (7.02 g, 50.770 mmol), and KI (20 mg) were added. The mixture was stirred at 85°C overnight. 100 mL of water and 100 mL of EA were added. After separation, the organic phase was washed with saturated brine, dried over anhydrous sodium sulfate, and then dried by column chromatography. Column chromatography was performed using DCM (containing 5% EA):MeOH = 50:1 to 5:1 to give 2.14 g of the product as a light yellow oil, with a yield of 53.6%. m / z [m+H + ]:747.26

[0296] Example 7 Synthesis of 3-(bis(4-(((Z)-octadec-9-en-1-yl)oxy)butyl)amino)propan-1-ol

[0297] Step 1: Synthesis of (Z)-1-(4-bromobutoxy)octadec-9-ene

[0298] 4-bromobutan-1-ol (5 g, 32.68 mmol) and (Z)-octadec-9-en-1-ol (9.65 g, 35.94 mmol) were mixed and heated to 120°C. After the reaction mixture was completely melted, 3 drops of concentrated sulfuric acid were added and stirring continued at 120°C overnight. After the reaction was completed, heating was stopped and 100 mL of EA and 100 mL of saturated NaHCO3 solution were added for washing. The aqueous phase was removed and the organic phase was washed with saturated brine, dried over anhydrous sodium sulfate, and spin-dried through a column. Column chromatography was performed using PE:EA = 10:1 as the eluent to obtain 9.89 g of the product as a colorless oil in a 75% yield.

[0299] Step 2: Synthesis of 3-(bis(4-(((Z)-octadec-9-en-1-yl)oxy)butyl)amino)propan-1-ol

[0300] Propanolamine (500 mg, 6.66 mmol) was dissolved in 50 mL of MeCN, and (Z)-1-(4-bromobutoxy)octadec-9-ene (6.71 g, 16.64 mmol), K2CO3 (5 g, 36.2 mmol), and KI (20 mg) were added. The mixture was stirred at 85°C overnight. 100 mL of water and 100 mL of EA were added. After separation, the organic phase was washed with saturated brine, dried over anhydrous sodium sulfate, and then dried by column chromatography. Column chromatography was performed using DCM (containing 5% EA):MeOH (containing 10% NH3) = 50:1 to 5:1 to give 2.3 g of the product as a light yellow oil in a yield of 47.9%. m / z [m+H + ]:721.26.

[0301] Example 8

[0302] Synthesis of (10Z,10'Z)-N,N'-(((2-hydroxyethyl)azanediyl)bis(propane-3,1-diyl))bis(nonadec-10-enamide)

[0303] The first step is the synthesis of di-tert-butyl(((2-hydroxyethyl)azanediyl)bis(propane-3,1-diyl))dicarbamate

[0304] Di-tert-butyl(azanediylbis(propane-3,1-diyl))dicarbamate (1 g, 3.02 mmol) and 2-bromoethan-1-ol (0.754 g, 6.03 mmol) were mixed and dissolved in 30 mL of acetonitrile. K2CO3 (1.65 g, 12 mmol) and KI (10 mg) were added and stirred at 85°C overnight. 100 mL of water and 100 mL of EA were added. After separation, the organic phase was washed with saturated brine, dried over anhydrous sodium sulfate, and then dried by column chromatography. Column chromatography was performed using DCM (containing 5% EA):MeOH (containing 10% NH3) in a ratio of 50:1 to 5:1 to afford 0.32 g of the product as a light yellow oil in a yield of 28.3%. m / z [m+H + ]:376.5.

[0305] Step 2: Synthesis of 2-(bis(3-aminopropyl)amino)ethan-1-ol

[0306] Dissolve 0.32g of di-tert-butyl(((2-hydroxyethyl)azanediyl)bis(propane-3,1-diyl))dicarbamate in 20mL of DCM and add 5mL of trifluoroacetic acid. After half an hour, TLC confirms complete reaction. The product is then spin-dried and dissolved in 100mL of EA. The organic layer is washed with saturated NaHCO3 solution and saturated brine, dried over anhydrous sodium sulfate, and then filtered through a column. Elution with DCM (containing 5% EA):MeOH (containing 10% NH3) in a ratio of 50:1 to 5:1 yields 0.05g of the product as a colorless oil in a 33.4% yield.

[0307] Step 3: Synthesis of (10Z,10'Z)-N,N'-(((2-hydroxyethyl)azanediyl)bis(propane-3,1-diyl))bis(nonadec-10-enamide)

[0308] 2-(bis(3-aminopropyl)amino)ethan-1-ol (50 mg, 0.285 mmol) and oleic acid (211 mg, 0.73 mmol) were mixed and 100 mL of DMF and a small amount of molecular sieves were added. After the reaction mixture was completely dissolved, 0.1 eq of DMAP and 2 eq of DCC were added and stirred overnight. After the reaction was completed, 100 mL of EA and 100 mL of saturated NaHCO3 solution were added for washing. The aqueous phase was removed, and the organic phase was washed with saturated brine, dried over anhydrous sodium sulfate, and dried by column chromatography. Column chromatography was performed using PE:EA = 10:1 to obtain 120 mg of the product as a colorless oil with a yield of 57%. m / z [m+H + ]:733.24

[0309] Example 9

[0310] (10Z,10'Z)-N,N'-(((3-hydroxypropyl)azanediyl)bis(propane-3,1-diyl))bis(nonadec-10-enamide) Synthesis

[0311] Step 1: Synthesis of di-tert-butyl(((3-hydroxypropyl)azanediyl)bis(propane-3,1-diyl))dicarbamate

[0312] Mix di-tert-butyl(azanediylbis(propane-3,1-diyl))dicarbamate (5 g, 15.08 mmol) and 3-bromopropan-1-ol (4.19 g, 30.17 mmol), dissolve in 50 mL of acetonitrile, add K2CO3 (10 g, 72 mmol) and KI (20 mg), and stir at 85°C overnight. Add 100 mL of water and 100 mL of EA, separate the liquid, and wash with saturated brine.

[0313] The organic phase was dried over anhydrous sodium sulfate and then dried by column chromatography. Column chromatography was performed using DCM (containing 5% EA): MeOH (containing 10% NH3) = 50:1 to 5:1 to give 2.47 g of a light yellow oily product in a yield of 42%. m / z [m+H + ]:390.5.

[0314] Step 2: Synthesis of 3-(bis(3-aminopropyl)amino)propan-1-ol

[0315] Dissolve 2.47g of di-tert-butyl(((3-hydroxypropyl)azanediyl)bis(propane-3,1-diyl))dicarbamate in 50mL of DCM and add 10mL of trifluoroacetic acid. After half an hour, TLC confirms complete reaction. The product is then spin-dried and dissolved in 100mL of EA. The organic layer is washed with saturated NaHCO3 solution and saturated brine, dried over anhydrous sodium sulfate, and then filtered through a column. Elution with DCM (containing 5% EA):MeOH (containing 10% NH3) in a ratio of 50:1 to 5:1 yields 0.97g of the product as a colorless oil in an 80.8% yield.

[0316] Step 3: Synthesis of (10Z,10'Z)-N,N'-(((3-hydroxypropyl)azanediyl)bis(propane-3,1-diyl))bis(nonadec-10-enamide)

[0317] 3-(bis(3-aminopropyl)amino)propan-1-ol (500 mg, 2.64 mmol) and oleic acid (1.96 g, 6.6 mmol) were mixed and 100 mL of DMF and a small amount of molecular sieves were added. After the reaction mixture was completely dissolved, 0.1 eq of DMAP and 2 eq of DCC were added and stirred overnight. After the reaction was completed, 100 mL of EA and 100 mL of a saturated NaHCO3 solution were added for washing. The aqueous phase was removed, and the organic phase was washed with saturated brine, dried over anhydrous sodium sulfate, and dried by column chromatography. Column chromatography was performed using PE:EA = 10:1 to obtain 380 mg of the product as a colorless oil with a yield of 19.28%. m / z [m+H + ]:747.26

[0318] Example 10 Synthesis of ((3-hydroxypropyl)azanediyl)bis(propane-3,1-diyl)(10Z,10'Z)-bis(nonadec-10-enoate)

[0319] Step 1: Synthesis of 3-bromopropyl(Z)-nonadec-10-enoate

[0320] 1-Bromobutanol (5.41g, 38.94mmol) and oleic acid (10g, 35.4mmol) were mixed and heated to 65°C. After the reaction mixture was completely melted, 3 drops of concentrated sulfuric acid were added and stirring continued at 65°C overnight. After the reaction was completed, heating was stopped and 100mL of EA and 100mL of NaHCO3 saturated solution were added for washing. The aqueous phase was removed and the organic phase was washed with saturated salt water and then dried over anhydrous sodium sulfate and spin-dried for column. Column chromatography was eluted with PE:EA=10:1 to give 11.5g of the product as a colorless oil with a yield of 77.8%.

[0321] Step 2: Synthesis of ((3-hydroxypropyl)azanediyl)bis(propane-3,1-diyl)(10Z,10'Z)-bis(nonadec-10-enoate)

[0322] Propanolamine (500 mg, 6.66 mmol) was dissolved in 50 mL of MeCN. 3-bromopropyl(Z)-nonadec-10-enoate (8.34 g, 19.97 mmol), K2CO3 (7.02 g, 50.770 mmol), and KI (20 mg) were added and stirred at 85°C overnight. 100 mL of water and 100 mL of EA were added. After separation, the organic phase was washed with saturated brine, dried over anhydrous sodium sulfate, and then dried by column chromatography. Column chromatography was performed using DCM (containing 5% EA):MeOH = 50:1 to 5:1 to obtain 1.78 g of the product as a light yellow oil in a yield of 35.7%. m / z [m+H + ]:749.23

[0323] Example 11 Assembly of liposome nanoparticles (LNP)

[0324] Liposome nanoparticles were formulated using the following ratios: cationic lipid: DOPE: chol: DMG-PEG = 50:10:38.75:1.25, with an N / P ratio of 6. Ionizable cationic lipid, DOPE, cholesterol, and DMG-PEG were prepared in 0.05 mmol / mL ethanol solution according to the above ratios. Subsequently, a 240 μg / mL solution of eGFP mRNA (molecular weight calculated as 320) in Tris-HCl buffer was prepared. Liposome nanoparticles were prepared using a microfluidic device at a volume ratio of 3 (water solution): 1 (ethanol solution) and a flow rate of 1:3. A 14 kD dialysis bag was used to exchange the solution with DPBS, resulting in liposome nanoparticles stored in DPBS. The mRNA content was determined using a nanodrop, and the particle size, PDI, and potential were measured using a zeta potential analyzer.

[0325] Example 12: Transfection of DC2.4 cells with eGFP mRNA using liposome nanoparticles (LNPs)

[0326] Dendritic cells play a crucial role in autoimmunity and anti-tumor immunity. DC2.4 is a dendritic cell lineage, and previous studies have shown that it expresses CD36. (H. Ma, W. Fang, Q. Li, Y. Wang, S. X. Hou, Arf1 Ablation in Colorectal Cancer Cells Activates a Super Signal Complex in DC to Enhance Anti-Tumor Immunity. Adv. Sci. 2023, 10, 2305089.) 2E5 cells were plated per well in a 24-well plate. DMEM medium supplemented with 10% FBS and 2.5% penicillin-streptomycin was used for incubation at 37°C in a CO2 incubator for 24 hours. Before transfection, the medium was aspirated, the cells were washed once with DPBS, and then serum-reduced Opti-MEM medium was added for 30 minutes. Subsequently, 200 ng of LNP001 containing eGFP mRNA prepared in Example 11 was added to each well and incubated overnight at 37°C. On the next day, the 24-well plate was directly photographed for fluorescence, and then digested normally. Cells were counted using trypan blue staining, and the fluorescence value was measured by flow cytometry.

[0327] Results: MFI values ​​ranged from 120,000 to 160,000, the positive rate was 95% to 98%, and the cell recovery rate was 56% to 84%.

[0328] Example 13: Transfection of A549 cells with eGFP mRNA using the liposome nanoparticles (LNPs):

[0329] A549 cells are a lung cancer cell line. Previous studies have shown that they express CD36 and are closely associated with cellular metabolism (Liu, H., Guo, W., Wang, T. et al. CD36 inhibition reduces non-small-cell lung cancer development through the AKT-mTOR pathway. Cell Biol Toxicol 40, 10). 2E5 cells were plated per well in a 24-well plate. The cells were cultured in REMI medium supplemented with 10% FBS and 2.5% penicillin-streptomycin at 37°C in a CO2 incubator for 24 hours. Before transfection, the medium was aspirated, the cells were washed once with DPBS, and then serum-reduced Opti-MEM medium was added for 30 minutes. Subsequently, LNPs containing 200 ng of eGFP-mRNA were added to each well and incubated at 37°C overnight. The next day, the 24-well plate was directly photographed for fluorescence, digested as usual, and cells were counted using trypan blue staining and fluorescence was measured by flow cytometry.

[0330] Results: MFI values ​​ranged from 89,000 to 110,000, the positive rate was 95% to 98%, and the cell recovery rate was 51% to 82%.

[0331] Example 14: In vivo effect evaluation experiment of liposome nanoparticles (LNP) in mice:

[0332] On day 0, Balb / c mice were randomly divided into 7 groups according to body weight: Note: Vehicle group: 10 mM Tris-HCl pH = 7.2 buffer solution

[0333] After grouping, drug administration began on the first day. Group reference 1, Group reference 2, Group A, Group B, Group C, Group D and Group E were administered by tail vein injection / intramuscular injection / subcutaneous injection, respectively. All were single doses, and the drug was administered once in total.

[0334] Results: Six hours after the start of drug administration, the whole-body fluorescence imaging of mice in Group Reference 1 and Group E was performed, and then the animals were euthanized for fluorescence imaging of the heart, liver, spleen, lung, kidney and lymph nodes. The whole-body bioluminescence value of the tail vein administration group of Group Reference 1 was 7.62E+05p / s; the bioluminescence values ​​of the heart, liver, spleen, lung, kidney and lymph nodes of Group Reference 1 were 2.19E+04p / s, 2.44E+04p / s, 1.55E+04p / s, 1.24E+04p / s, 1.84E+04p / s and 1.82E+04p / s, respectively; the whole-body bioluminescence value of the tail vein administration group of Group E was 2.12E+07±2.27E+06p / s; the bioluminescence values ​​of the heart, liver, spleen, lung, kidney and lymph nodes of Group E were 1.9 The expression of the LNPs in the lymph nodes of groups 1 and E was 5E+04±2.58E+03p / s, 1.04E+06±5.23E+04p / s, 1.42E+06±6.35E+05p / s, 3.32E+06±1.47E+06p / s, 4.55E+04±3.19E+03p / s, and 1.45E+04±1.95E+03p / s, respectively. The %eGFP+ / mCD45+ ratios in the lymph nodes of groups 1 and E were 0.25% and 0.17%, respectively. The %eGFP+ / mCD45+ ratios in the spleen of groups 1 and E were 1.72% and 1.95%, respectively. These results indicate that the LNPs were primarily localized in tissues such as the liver, spleen, and lungs after tail vein administration, and that they enhanced macrophage transfection efficiency in the spleen.

[0335] Fluorescence imaging was performed on mice in Group Reference 1, Group Reference 2, Group A, Group B, Group C, Group D, and Group E 6, 24, and 48 hours after dosing (fluorescence values ​​at the intramuscular injection site and liver can be compared between Reference 2 and Group A / B). Because bioluminescence values ​​for subcutaneous-administered mice in all groups were less than 1E7 24 hours after dosing, fluorescence imaging was performed only on mice injected intramuscularly and via the tail vein in each group at 48 hours. The overall bioluminescence value of the tail vein administration group of reference group 1 was 9.34E+05p / s; the overall bioluminescence value of the muscle administration group of reference group 1 was 8.03E+05p / s; the overall bioluminescence value of the muscle administration group of reference group 2 was 7.46E+09±2.77E+09p / s, the liver luminescence value of the muscle administration group of reference group 2 was 4.90E+09±2.08E+09p / s, the left leg muscle tissue luminescence value of reference group 2 was 1.02E+09±2.67E+08p / s, and the right leg muscle tissue luminescence value of reference group 2 was 1.16E+0 9±3.82E+08p / s; the overall bioluminescence value of the tail vein administration group of group A was 1.66E+08±2.16E+06p / s; the overall bioluminescence value of the muscle administration group of group A was 2.54E+08±4.4E+06p / s, the liver luminescence value of group A was 4.87E+07±1.44E+07p / s, the luminescence value of the left leg muscle tissue of group A was 1.23E+08±5.90E+06p / s, and the luminescence value of the right leg muscle tissue of group A was 6.96E+07±3.21E+06p / s; group A The overall bioluminescence value of the subcutaneous administration group was 2.14E+07±7.88E+06; the overall bioluminescence value of the tail vein administration group of group B was 1.75E+08±1.67E+07p / s; the overall bioluminescence value of the intramuscular administration group of group B was 1.19E+08±2.76E+07p / s, the intramuscular administration liver luminescence value of group B was 9.54E+06±1.95E+06p / s, the intramuscular administration left leg muscle tissue luminescence value of group B was 2.97E+07±1.87E+06p / s, and the intramuscular administration right leg muscle tissue luminescence value of group B was 7.19E+ 07±2.83E+07p / s; the overall bioluminescence value of the subcutaneous administration group in group B was 5.47E+06±1.52E+06; the overall bioluminescence value of the tail vein administration group in group C was 3.52E+07±4.43E+06p / s; the overall bioluminescence value of the intramuscular administration group in group C was 1.92E+08±8.87E+07p / s; the overall bioluminescence value of the tail vein administration group in group D was 3.12E+06±1.11E+06p / s; the overall bioluminescence value of the intramuscular administration group in group D was 1.82E+07±1.13E+06p / s.The overall bioluminescence value of the tail vein administration group in group E was 1.21E+07±6.03E+06p / s; the overall bioluminescence value of the muscle administration group in group E was 1.79E+07±8.22E+06p / s.

[0336] Fluorescence data from mice preliminarily indicated that the model Flux mRNA was successfully delivered into mice by LNP and was able to efficiently express fluorescent protein. Compared with the commercial reference 2LNP, relatively fewer off-target LNPs were distributed to the liver after intramuscular injection, and the muscle:liver fluorescence was relatively higher, reflecting superior tissue-specific delivery characteristics.

[0337] Example 15: Targeted Muscle CRISPR-Mediated Gene Editing Using the Lipid Nanoparticles

[0338] LNP001 or Lipo3000 (purchased from Thermo Fisher) was mixed with an H11-targeting sgRNA and Cas-9 mRNA at a lipid:RNA ratio of 20:1. After dialysis and concentration, the mixture was injected intramuscularly into the calf gastrocnemius muscle of C57BL / 6J mice at a dose of 2-10 ml / kg. Four days later, the mice were euthanized and their gastrocnemius muscle tissue was isolated. RNA was then extracted using a Qiagen kit and reverse transcribed into cDNA. The gene editing efficiency of the model CRISPR system delivered by different lipid nanoparticles was tested by quantitative PCR using the designed primers 5'-TGGATTTTGACTGCAGGGGTAAA-3' (SEQ ID NO. 1) and 5'-CTGTGGCTTTGGAGCCTACACTG-3' (SEQ ID NO. 2). The results are shown in Figure 4. Compared with the non-CD36-targeting nanocomplex group Lipo3000, the targeted LNP001 group significantly improved the gene editing efficiency in the target tissue.

[0339] Example 16: Targeted drug delivery of anti-tumor PROTAC using the lipid nanoparticles

[0340] ARV-771 (purchased from MedChem Express) is a PROTAC molecule targeting BRD4. ARV-771 was first dissolved in DMSO and then dissolved in PBS to a concentration of 100 nM. Lipo2000 or LNP001 was added to the ARV-771 to a final concentration of 5 μg / ml and then incubated for 15 minutes. 6Lipo2000-AR-771 or LPN001-ARV-771 was added to a six-well plate of Hela cells and cultured for 24 hours. Protein was extracted and the target protein degradation was evaluated by Western blot. The results are shown in Figure 5. The target protein degradation level of LPN001-ARV-771 was significantly higher than that of AR-771. Compared with the non-CD36-targeted nanocomplex group Lipo2000-AR-771, LPN001-ARV-771 carrying a PROTAC molecule targeting BRD4 significantly improved the degradation efficiency of Hela cell BRD4 induced by AR-771.

Claims

1. A compound of formula (I): in, R 1 independently selected from the group consisting of hydroxy-substituted alkyl, hydroxy-substituted alkenyl, and hydroxy-substituted alkynyl; M 1 and M 2 are each independently selected from the group consisting of optionally substituted alkylene, optionally substituted alkenylene, and optionally substituted alkynylene; L 1 and L 2 are each independently selected from the group consisting of -C(O)O-, -OC(O)-, -O-, -C(O)N(R)-, and -N(R)C(O)-, wherein each R is independently selected from the group consisting of: C 1-3 Alkyl, C 2-3 alkenyl and H; M 3 and M 4 Each independently where R 2 Independently selected from linear C 4-10 Alkylene, straight chain C 4-10 Alkenylene and straight chain C 4-10 The group consisting of alkynylene, R 3 Independently selected from linear C 4-10 Alkyl, straight chain C 4-10 Alkenyl and straight chain C 4-10 A group consisting of alkynyl groups.

2. The compound according to claim 1, wherein R 2 Independently selected from linear C 6-9 Alkylene, straight chain C 6-9 Alkenylene and straight chain C 6-9 The group consisting of alkynylene, R 3 Independently selected from linear C 6-9 Alkyl, straight chain C 6-9 Alkenyl and straight chain C 6-9 A group consisting of alkynyl groups.

3. The compound according to any one of claims 1 to 2, wherein R 2 Independently linear C 6-9 Alkylene or linear C 6-9 Alkenylene, R 3 Independently linear C 6-9 Alkyl or linear C 6-9 Alkenyl.

4. The compound according to any one of claims 1 to 3, wherein R 2 and R 3 They are not all unsaturated hydrocarbon groups.

5. The compound according to any one of claims 1 to 4, wherein M 3 and M 4 Each independently a linear C 10-20 Alkenyl or straight chain C 10-20 Alkynyl.

6. The compound according to any one of claims 1 to 5, wherein M 3 and M 4 Each independently a linear C 14-18 Alkenyl or straight chain C 14-18 Alkynyl.

7. The compound according to any one of claims 1 to 6, wherein M 3 and M 4 Each independently a linear C 18 Alkenyl or straight chain C 18 Alkynyl.

8. The compound according to any one of claims 1 to 7, wherein M 3 and M 4 For straight chain C 18 Alkenyl.

9. The compound according to any one of claims 1 to 8, wherein R 1 independently selected from hydroxy substituted C 1-6 Alkyl, hydroxy substituted C 2-6 Alkenyl and hydroxy substituted C 2-6 A group consisting of alkynyl groups.

10. The compound according to any one of claims 1 to 9, wherein R 1 C containing hydroxyl substitution 1-6 alkyl.

11. The compound according to any one of claims 1 to 10, wherein R 1 Selected from the group consisting of -CH2OH, -(CH2)2OH, -(CH2)3OH, -(CH2)4OH, -(CH2)5OH and -(CH2)6OH.

12. The compound according to any one of claims 1 to 11, wherein M 1 and M 2 are each independently an optionally substituted C 1-4 Alkylene, and when M 1 and M 2 When R is C2 alkylene 1 Not -(CH2)6OH.

13. The compound according to any one of claims 1 to 12, wherein M 1 and M 2 are each independently an optionally substituted C 3-4 Alkylene.

14. The compound according to any one of claims 1 to 13, wherein the compound has formula (II): in, m is 2, 3, 4 or 5; Each n is independently 3 or 4; L 1 and L 2 each independently selected from the group consisting of -C(O)O-, -OC(O)-, -O-, -C(O)N(H)-, and -N(H)C(O)-; M 3 and M 4 Each independently where R 2 Independently selected from linear C 4-10 Alkylene, straight chain C 4-10 Alkenylene and straight chain C 4-10 The group consisting of alkynylene, R 3 Independently selected from linear C 4-10 Alkyl, straight chain C 4-10 Alkenyl and straight chain C 4-10 A group consisting of alkynyl groups.

15. The compound according to any one of claims 1 to 14, wherein R 2 Independently selected from linear C 6-9 Alkylene, straight chain C 6-9 Alkenylene and straight chain C 6-9 The group consisting of alkynylene, R 3 Independently selected from linear C 6-9 Alkyl, straight chain C 6-9 Alkenyl and straight chain C 6-9 A group consisting of alkynyl groups.

16. A compound according to any one of claims 1 to 15, wherein R 2 Independently linear C 6-9 Alkylene or linear C 6-9 Alkenylene, R 3 Independently linear C 6-9 Alkyl or linear C 6-9 Alkenyl.

17. A compound according to any one of claims 1 to 16, wherein R 2 and R 3 They are not all unsaturated hydrocarbon groups.

18. A compound according to any one of claims 1 to 17, wherein M 3 and M 4 Each independently a linear C 10-20 Alkenyl or straight chain C 10-20 Alkynyl.

19. The compound according to any one of claims 1 to 18, wherein M 3 and M 4 Each independently a linear C 14-18 Alkenyl or straight chain C 14-18 Alkynyl.

20. The compound according to any one of claims 1 to 19, wherein M 3 and M 4 Each independently a linear C 18 Alkenyl or straight chain C 18 Alkynyl.

21. The compound according to any one of claims 1 to 20, wherein M 3 and M 4 For straight chain C 18 Alkenyl.

22. The compound according to any one of claims 1 to 21, wherein the compound has formula (III): in, m is 2, 3 or 4; L 1 and L 2 each independently selected from the group consisting of -C(O)O-, -OC(O)-, -CO-, -C(O)N(H)-, and -N(H)C(O)-; M 3 and M 4 Each independently where R 2 Independently linear C 4-10 Alkylene or linear C 4-10 Alkenylene, R 3 Independently linear C 6-9 Alkyl or linear C 4-10 Alkenyl.

23. A compound according to any one of claims 1 to 22, wherein R 2 Independently linear C 6-9 Alkylene or linear C 6-9 Alkenylene, R 3 Independently linear C 6-9 Alkyl or linear C 6-9 Alkenyl.

24. A compound according to any one of claims 1 to 23, wherein R 2 and R 3 They are not all unsaturated hydrocarbon groups.

25. A compound according to any one of claims 1 to 24, wherein R 2 and R 3 Each independently a linear C 6-9 Alkylene.

26. A compound according to any one of claims 1 to 25, wherein M 3 and M 4 For straight chain C 10-20 Alkenyl or straight chain C 10-20 Alkynyl.

27. A compound according to any one of claims 1 to 26, wherein M 3 and M 4 For straight chain C 14-18 Alkenyl or straight chain C 14-18 Alkynyl.

28. A compound according to any one of claims 1 to 27, wherein M 3 and M 4 For straight chain C 18 Alkenyl.

29. The compound according to any one of claims 1 to 28, wherein M 3 and M 4 Each independently selected from:

30. A compound according to any one of claims 1 to 29, wherein M 3 and M 4 for 31. A compound of formula (IV): in, m is 2 or 3; L 1 and L 2 are each independently selected from the group consisting of -C(O)O-, -OC(O)-, -CO-, -C(O)N(H)- and -N(H)C(O)-.

32. A compound according to any one of claims 1 to 31, wherein L 1 and L 2 Each is independently selected from -C(O)O-, -OC(O)-, -O-, -C(O)N(H)- and -N(H)C(O)-.

33. A compound according to any one of claims 1 to 32, wherein L 1 and L 2 It is -C(O)O-.

34. A compound according to any one of claims 1 to 32, wherein L 1 and L 2 It is -OC(O)-.

35. A compound according to any one of claims 1 to 32, wherein L 1 and L 2 For -CO-.

36. A compound according to any one of claims 1 to 32, wherein L 1 and L 2 It is -C(O)N(H)-.

37. A compound according to any one of claims 1 to 32, wherein L 1 and L 2 It is -N(H)C(O)-.

38. A compound selected from the group consisting of:

39. A nanoparticle composition comprising a lipid component of a compound according to any one of claims 1-38.

40. The nanoparticle composition of claim 39, wherein the lipid component further comprises a phospholipid.

41. The nanoparticle composition of any one of claims 39-40, wherein the phospholipid is selected from the group consisting of: 1,2-Dilinoleoyl-sn-glycero-3-phosphocholine (DLPC), 1,2-Dimyristoyl-sn-glycero-phosphocholine (DMPC), 1,2-Dioleoyl-sn-glycero-3-phosphocholine (DOPC), 1,2-Dipalmitoyl-sn-glycero-3-phosphocholine (DPPC), 1,2-Distearoyl-sn-glycero-3-phosphocholine (DSPC), 1,2-Dicondecanoyl-sn-glycero-phosphocholine (DUPC), 1-Palmitoyl-2-oleoyl-sn-glycero-3-phosphocholine (POPC), 1,2-Di-O-octadecenyl-sn-glycero-3-phosphocholine (18:0Diether PC), 1-oleoyl-2-cholesteryl hemisuccinyl-sn-glycero-3-phosphocholine (OChemsPC), 1-hexadecyl-sn-glycero-3-phosphocholine (C16 Lyso PC), 1,2-dialinolenoyl-sn-glycero-3-phosphocholine, 1,2-diarachidonoyl-sn-glycero-3-phosphocholine, 1,2-didocosahexaenoyl-sn-glycero-3-phosphocholine, 1,2-dioleoyl-sn-glycero-3-phosphoethanolamine (DOPE), 1,2-diphytanoyl-sn-glycero-3-phosphoethanolamine (ME 16.0PE), 1,2-distearoyl-sn-glycero-3-phosphoethanolamine, 1,2-dilinoleoyl-sn-glycero-3-phosphoethanolamine, 1,2-dilinolenoyl-sn-glycero-3-phosphoethanolamine, 1,2-diarachidonoyl-sn-glycero-3-phosphoethanolamine, 1,2-bisdocosahexaenoyl-sn-glycero-3-phosphoethanolamine, 1,2-dioleoyl-sn-glycero-3-phospho-rac-(1-glycerol) sodium salt (DOPG), dipalmitoylphosphatidylglycerol (DPPG), palmitoyloleoyl phosphatidylethanolamine (POPE), distearoyl-phosphatidyl-ethanolamine (DSPE), dipalmitoylphosphatidylethanolamine (DPPE), dimyristoylphosphoethanolamine (DMPE), 1-stearoyl-2-oleoyl-stearoylethanolamine (SOPE), 1-stearoyl-2-oleoyl-phosphatidylcholine (SOPC), sphingomyelin, phosphatidylcholine, phosphatidylethanolamine, phosphatidylserine, phosphatidylinositol, phosphatidic acid, palmitoyloleoylphosphatidylcholine, lysophosphatidylcholine, lysophosphatidylethanolamine (LPE), and mixtures thereof.

42. The nanoparticle composition of any one of claims 39-42, wherein the phospholipid is DOPE.

43. The nanoparticle composition of any one of claims 39-42, wherein the phospholipid is DSPC.

44. The nanoparticle composition of any one of claims 39-43, wherein the lipid component further comprises a structural lipid.

45. The nanoparticle composition of any one of claims 39-44, wherein the structured lipid is selected from the group consisting of cholesterol, coproposterol, sitosterol, ergosterol, campesterol, stigmasterol, brassicasterol, tomatidine, ursolic acid, alpha-tocopherol, and mixtures thereof.

46. ​​The nanoparticle composition of any one of claims 39-45, wherein the structural lipid is cholesterol.

47. The nanoparticle composition of any one of claims 39-46, wherein the lipid component further comprises a PEG lipid.

48. The nanoparticle composition of any one of claims 39-47, wherein the PEG lipid is selected from the group consisting of PEG-modified phosphatidylethanolamine, PEG-modified phosphatidic acid, PEG-modified ceramide, PEG-modified dialkylamine, PEG-modified diacylglycerol, PEG-modified dialkylglycerol, and mixtures thereof.

49. The nanoparticle composition of any one of claims 39-48, wherein the PEG lipid comprises a PEG moiety ranging in size from about 1000 Da to about 20 kDa.

50. The nanoparticle composition of any one of claims 39-49, wherein the PEG lipid is selected from any one or a combination of more than one of PEG1000-DMG, PEG5000-DMG, PEG2000-DMG and PEG2000-DSPE.

51. The nanoparticle composition of any one of claims 39-50, wherein the PEG lipid is PEG2000-DMG.

52. The nanoparticle composition of any one of claims 39-51, wherein the lipid component further comprises phospholipids, structural lipids, and PEG lipids.

53. The nanoparticle composition of any one of claims 39-52, wherein the lipid component comprises about 30 mol% to about 60 mol% of the compound, about 0 mol% to about 30 mol% phospholipids, about 18.5 mol% to about 48.5 mol% structural lipids, and about 0 mol% to about 10 mol% PEG lipids.

54. The nanoparticle composition of any one of claims 39-53, wherein the lipid component comprises about 50 mol% of the compound, about 10 mol% phospholipids, about 38.5 mol% structural lipids, and about 1.5 mol% PEG lipids.

55. The nanoparticle composition of any one of claims 39-54, wherein the lipid component comprises about 50 mol% of Compound 1 or Compound 2, about 10 mol% of DOPE or DSPC, about 37.0-39.5 mol% of cholesterol or sitosterol, and about 0.5-3.0 mol% of PEG2000-DMG.

56. The nanoparticle composition of any one of claims 39-55, further comprising a therapeutic and / or prophylactic agent.

57. The nanoparticle composition of claim 56, wherein the therapeutic and / or prophylactic agent is an anticancer agent, an antiviral agent, an immunomodulatory agent, an anti-inflammatory agent, or an agent that modulates cellular metabolic activity.

58. The nanoparticle composition of any one of claims 56-57, wherein the therapeutic and / or prophylactic agent is a nucleic acid, a protein, a peptide, or a small molecule.

59. The nanoparticle composition of any one of claims 56-58, wherein the therapeutic and / or prophylactic agent is a nucleic acid.

60. The nanoparticle composition of any one of claims 56-59, wherein the therapeutic and / or prophylactic agent is ribonucleic acid (RNA).

61. The nanoparticle composition of claim 60, wherein the RNA is selected from the group consisting of small interfering RNA (siRNA), asymmetric interfering RNA (aiRNA), microRNA (miRNA), Dicer-substrate RNA (dsRNA), small hairpin RNA (shRNA), messenger RNA (mRNA), and mixtures thereof.

62. The nanoparticle composition of any one of claims 60-61, wherein the RNA is mRNA.

63. The nanoparticle composition of any one of claims 61-62, wherein the mRNA comprises one or more of: a stem-loop, a chain-terminating nucleoside, a poly(A) sequence, a polyadenylation signal, and / or a 5' cap structure.

64. The nanoparticle composition of any one of claims 56-63, wherein the therapeutic and / or prophylactic agent comprises a component of a gene editing system.

65. The nanoparticle composition of claim 64, wherein components of the gene editing system comprise a polynucleotide encoding a nuclease.

66. The nanoparticle composition of claim 65, wherein the nuclease is selected from the group consisting of zinc finger nucleases (ZFNs), transcription activator-like effector nucleases (TALENs), CRISPR-associated protein 9 (Cas9), and engineered homing endonucleases.

67. The nanoparticle composition of any one of claims 65-66, wherein the nuclease is Cas9 and the nanoparticle composition further comprises a guide RNA that targets the nuclease to a specific site in the target cell genome.

68. The nanoparticle composition of any one of claims 64-67, wherein the components of the gene editing system include i) CRISPR-associated protein 9 (Cas9), or mRNA encoding CRISPR-associated protein 9 (Cas9); and ii) a guide RNA nucleic acid of a single guide RNA (sgRNA), or a guide RNA nucleic acid encoding a single guide RNA (sgRNA).

69. The nanoparticle composition of any one of claims 56-68, wherein the therapeutic and / or prophylactic agent comprises a small molecule.

70. The nanoparticle composition of any one of claims 56-69, wherein the therapeutic and / or prophylactic agent comprises a PROTAC (Proteolysis Targeting Chimera) molecule.

71. The nanoparticle composition of claim 70, wherein the PROTAC is selected from the group consisting of ARV-110, ARV-471, ARV-766, ARV-771, AVR-825, AR-LDD, DT2216, KT-474, KT-413, KT-333, NX-2127, NX-5948, CG001419, CFT8634, FHD-609, and SARD279.

72. The nanoparticle composition of any one of claims 56-71, wherein the wt / wt ratio of the lipid component to the therapeutic and / or prophylactic agent is about 10:1 to about 60:

1.

73. The nanoparticle composition of any one of claims 56-72, wherein the nanoparticle composition has an N:P ratio of about 2:1 to about 30:

1.

74. The nanoparticle composition of any one of claims 56-73, wherein the average size of the nanoparticle composition is about 70 nm to about 100 nm.

75. Use of the compound of any one of claims 1-38 or the nanoparticle composition of any one of claims 39-74 in preparing a delivery system.

76. The use according to claim 75, wherein the delivery system is a CD36-targeted delivery system.

77. A delivery system comprising the compound of any one of claims 1-38 or the nanoparticle composition of any one of claims 39-74.

78. The delivery system according to claim 77, wherein the delivery system is a CD36-targeted delivery system.

79. A pharmaceutical composition comprising the nanoparticle composition of any one of claims 39-74 and a pharmaceutically acceptable carrier.

80. A method of delivering a therapeutic and / or prophylactic agent to a mammalian cell, the method comprising administering to a subject in need thereof the nanoparticle composition of any one of claims 39-74, the administration comprising contacting the cell with the nanoparticle composition, thereby delivering the therapeutic and / or prophylactic agent to the cell.

81. The method of claim 80, wherein the cells express CD36.

82. The method of any one of claims 80-81, wherein the mammalian cell is in a mammal.

83. The method of any one of claims 80-82, wherein the mammal is a human.

84. A method of delivering a therapeutic and / or prophylactic agent to an organ or tissue, the method comprising contacting the organ or tissue with the nanoparticle composition according to any one of claims 39-74, thereby delivering the therapeutic and / or prophylactic agent to the cells.

85. The method of claim 84, wherein the organ or tissue is an organ or tissue that expresses CD36.

86. A method for preventing and / or treating a disease or condition, comprising administering to a subject in need thereof an effective amount of the nanoparticle composition according to any one of claims 39-74.

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