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WO2025195308A1 - Composés lipidiques cationiques, composition et utilisation associées - Google Patents

Composés lipidiques cationiques, composition et utilisation associées

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Publication number
WO2025195308A1
WO2025195308A1 PCT/CN2025/082780 CN2025082780W WO2025195308A1 WO 2025195308 A1 WO2025195308 A1 WO 2025195308A1 CN 2025082780 W CN2025082780 W CN 2025082780W WO 2025195308 A1 WO2025195308 A1 WO 2025195308A1
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linear
group
nanoparticle composition
independently
compound according
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Chinese (zh)
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胡天楠
黄奡
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    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07C—ACYCLIC OR CARBOCYCLIC COMPOUNDS
    • C07C229/00—Compounds containing amino and carboxyl groups bound to the same carbon skeleton
    • C07C229/02—Compounds containing amino and carboxyl groups bound to the same carbon skeleton having amino and carboxyl groups bound to acyclic carbon atoms of the same carbon skeleton
    • C07C229/04—Compounds containing amino and carboxyl groups bound to the same carbon skeleton having amino and carboxyl groups bound to acyclic carbon atoms of the same carbon skeleton the carbon skeleton being acyclic and saturated
    • C07C229/06—Compounds containing amino and carboxyl groups bound to the same carbon skeleton having amino and carboxyl groups bound to acyclic carbon atoms of the same carbon skeleton the carbon skeleton being acyclic and saturated having only one amino and one carboxyl group bound to the carbon skeleton
    • C07C229/10—Compounds containing amino and carboxyl groups bound to the same carbon skeleton having amino and carboxyl groups bound to acyclic carbon atoms of the same carbon skeleton the carbon skeleton being acyclic and saturated having only one amino and one carboxyl group bound to the carbon skeleton the nitrogen atom of the amino group being further bound to acyclic carbon atoms or to carbon atoms of rings other than six-membered aromatic rings
    • C07C229/16—Compounds containing amino and carboxyl groups bound to the same carbon skeleton having amino and carboxyl groups bound to acyclic carbon atoms of the same carbon skeleton the carbon skeleton being acyclic and saturated having only one amino and one carboxyl group bound to the carbon skeleton the nitrogen atom of the amino group being further bound to acyclic carbon atoms or to carbon atoms of rings other than six-membered aromatic rings to carbon atoms of hydrocarbon radicals substituted by amino or carboxyl groups, e.g. ethylenediamine-tetra-acetic acid, iminodiacetic acids
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K31/00—Medicinal preparations containing organic active ingredients
    • A61K31/70—Carbohydrates; Sugars; Derivatives thereof
    • A61K31/7088—Compounds having three or more nucleosides or nucleotides
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K48/00—Medicinal preparations containing genetic material which is inserted into cells of the living body to treat genetic diseases; Gene therapy
    • A61K48/0008—Medicinal preparations containing genetic material which is inserted into cells of the living body to treat genetic diseases; Gene therapy characterised by an aspect of the 'non-active' part of the composition delivered, e.g. wherein such 'non-active' part is not delivered simultaneously with the 'active' part of the composition
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K48/00—Medicinal preparations containing genetic material which is inserted into cells of the living body to treat genetic diseases; Gene therapy
    • A61K48/005—Medicinal preparations containing genetic material which is inserted into cells of the living body to treat genetic diseases; Gene therapy characterised by an aspect of the 'active' part of the composition delivered, i.e. the nucleic acid delivered
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K9/00—Medicinal preparations characterised by special physical form
    • A61K9/48—Preparations in capsules, e.g. of gelatin, of chocolate
    • A61K9/50—Microcapsules having a gas, liquid or semi-solid filling; Solid microparticles or pellets surrounded by a distinct coating layer, e.g. coated microspheres, coated drug crystals
    • A61K9/51—Nanocapsules; Nanoparticles
    • A61K9/5107—Excipients; Inactive ingredients
    • A61K9/5123—Organic compounds, e.g. fats, sugars
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P29/00—Non-central analgesic, antipyretic or antiinflammatory agents, e.g. antirheumatic agents; Non-steroidal antiinflammatory drugs [NSAID]
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P3/00—Drugs for disorders of the metabolism
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P31/00—Antiinfectives, i.e. antibiotics, antiseptics, chemotherapeutics
    • A61P31/12—Antivirals
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P35/00—Antineoplastic agents
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P37/00—Drugs for immunological or allergic disorders
    • A61P37/02—Immunomodulators
    • B—PERFORMING OPERATIONS; TRANSPORTING
    • B82—NANOTECHNOLOGY
    • B82Y—SPECIFIC USES OR APPLICATIONS OF NANOSTRUCTURES; MEASUREMENT OR ANALYSIS OF NANOSTRUCTURES; MANUFACTURE OR TREATMENT OF NANOSTRUCTURES
    • B82Y40/00—Manufacture or treatment of nanostructures
    • B—PERFORMING OPERATIONS; TRANSPORTING
    • B82—NANOTECHNOLOGY
    • B82Y—SPECIFIC USES OR APPLICATIONS OF NANOSTRUCTURES; MEASUREMENT OR ANALYSIS OF NANOSTRUCTURES; MANUFACTURE OR TREATMENT OF NANOSTRUCTURES
    • B82Y5/00—Nanobiotechnology or nanomedicine, e.g. protein engineering or drug delivery
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07C—ACYCLIC OR CARBOCYCLIC COMPOUNDS
    • C07C217/00—Compounds containing amino and etherified hydroxy groups bound to the same carbon skeleton
    • C07C217/02—Compounds containing amino and etherified hydroxy groups bound to the same carbon skeleton having etherified hydroxy groups and amino groups bound to acyclic carbon atoms of the same carbon skeleton
    • C07C217/04—Compounds containing amino and etherified hydroxy groups bound to the same carbon skeleton having etherified hydroxy groups and amino groups bound to acyclic carbon atoms of the same carbon skeleton the carbon skeleton being acyclic and saturated
    • C07C217/06—Compounds containing amino and etherified hydroxy groups bound to the same carbon skeleton having etherified hydroxy groups and amino groups bound to acyclic carbon atoms of the same carbon skeleton the carbon skeleton being acyclic and saturated having only one etherified hydroxy group and one amino group bound to the carbon skeleton, which is not further substituted
    • C07C217/08—Compounds containing amino and etherified hydroxy groups bound to the same carbon skeleton having etherified hydroxy groups and amino groups bound to acyclic carbon atoms of the same carbon skeleton the carbon skeleton being acyclic and saturated having only one etherified hydroxy group and one amino group bound to the carbon skeleton, which is not further substituted the oxygen atom of the etherified hydroxy group being further bound to an acyclic carbon atom
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07C—ACYCLIC OR CARBOCYCLIC COMPOUNDS
    • C07C219/00—Compounds containing amino and esterified hydroxy groups bound to the same carbon skeleton
    • C07C219/02—Compounds containing amino and esterified hydroxy groups bound to the same carbon skeleton having esterified hydroxy groups and amino groups bound to acyclic carbon atoms of the same carbon skeleton
    • C07C219/04—Compounds containing amino and esterified hydroxy groups bound to the same carbon skeleton having esterified hydroxy groups and amino groups bound to acyclic carbon atoms of the same carbon skeleton the carbon skeleton being acyclic and saturated
    • C07C219/08—Compounds containing amino and esterified hydroxy groups bound to the same carbon skeleton having esterified hydroxy groups and amino groups bound to acyclic carbon atoms of the same carbon skeleton the carbon skeleton being acyclic and saturated having at least one of the hydroxy groups esterified by a carboxylic acid having the esterifying carboxyl group bound to an acyclic carbon atom of an acyclic unsaturated carbon skeleton
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07C—ACYCLIC OR CARBOCYCLIC COMPOUNDS
    • C07C233/00—Carboxylic acid amides
    • C07C233/01—Carboxylic acid amides having carbon atoms of carboxamide groups bound to hydrogen atoms or to acyclic carbon atoms
    • C07C233/34—Carboxylic acid amides having carbon atoms of carboxamide groups bound to hydrogen atoms or to acyclic carbon atoms having the nitrogen atom of at least one of the carboxamide groups bound to a carbon atom of a hydrocarbon radical substituted by amino groups
    • C07C233/35—Carboxylic acid amides having carbon atoms of carboxamide groups bound to hydrogen atoms or to acyclic carbon atoms having the nitrogen atom of at least one of the carboxamide groups bound to a carbon atom of a hydrocarbon radical substituted by amino groups with the substituted hydrocarbon radical bound to the nitrogen atom of the carboxamide group by an acyclic carbon atom
    • C07C233/38—Carboxylic acid amides having carbon atoms of carboxamide groups bound to hydrogen atoms or to acyclic carbon atoms having the nitrogen atom of at least one of the carboxamide groups bound to a carbon atom of a hydrocarbon radical substituted by amino groups with the substituted hydrocarbon radical bound to the nitrogen atom of the carboxamide group by an acyclic carbon atom having the carbon atom of the carboxamide group bound to a carbon atom of an acyclic unsaturated carbon skeleton
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07C—ACYCLIC OR CARBOCYCLIC COMPOUNDS
    • C07C237/00—Carboxylic acid amides, the carbon skeleton of the acid part being further substituted by amino groups
    • C07C237/02—Carboxylic acid amides, the carbon skeleton of the acid part being further substituted by amino groups having the carbon atoms of the carboxamide groups bound to acyclic carbon atoms of the carbon skeleton
    • C07C237/04—Carboxylic acid amides, the carbon skeleton of the acid part being further substituted by amino groups having the carbon atoms of the carboxamide groups bound to acyclic carbon atoms of the carbon skeleton the carbon skeleton being acyclic and saturated
    • C07C237/12—Carboxylic acid amides, the carbon skeleton of the acid part being further substituted by amino groups having the carbon atoms of the carboxamide groups bound to acyclic carbon atoms of the carbon skeleton the carbon skeleton being acyclic and saturated having the nitrogen atom of at least one of the carboxamide groups bound to an acyclic carbon atom of a hydrocarbon radical substituted by carboxyl groups

Definitions

  • 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.
  • LNPs Liposomal nanoparticles
  • ionizable cationic lipids 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.
  • 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.
  • 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.
  • lipid nanoparticles have demonstrated their practicality as a delivery platform for mRNA vaccines and therapies.
  • ionizable cationic lipid molecules have 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).
  • hATTR transthyretin amyloidosis
  • Moderna and BioNTech/Pfizer have developed two COVID-19 vaccines using SM-102 and ALC-0315, respectively.
  • lipid nanoparticles such as protein degradation targeting chimeras (PROTACs), which have received much attention in recent years.
  • PROTACs protein degradation targeting chimeras
  • PROTACs are highly anticipated in anti-tumor treatment because they can achieve efficient degradation of previously difficult-to-drug targets.
  • 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.
  • 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.
  • DCs dendritic cells
  • MVECs microvascular endothelial cells
  • CD36 is expressed in tumor cells, stromal cells, and immune cells, but expression levels vary across cell types and tumor stages.
  • 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- ⁇ signaling pathway.
  • EMT epithelial-mesenchymal transition
  • 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.
  • Sulfosuccinimidyl oleate 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).
  • 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.
  • the present invention mainly provides the following technical solutions:
  • the present application provides a compound of formula (I):
  • R 1 is independently selected from the group consisting of hydroxy-substituted alkyl, hydroxy-substituted alkenyl, and hydroxy-substituted alkynyl;
  • M1 and M2 are each independently selected from the group consisting of optionally substituted alkylene, optionally substituted alkenylene and optionally substituted alkynylene;
  • L1 and L2 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 C1-3 alkyl, C2-3 alkenyl, and H;
  • M3 and M4 are each independently wherein R 2 is independently selected from the group consisting of linear C 4-10 alkylene, linear C 4-10 alkenylene and linear C 4-10 alkynylene, and R 3 is independently selected from the group consisting of linear C 4-10 alkyl, linear C 4-10 alkenyl and linear C 4-10 alkynyl.
  • R 2 is independently selected from the group consisting of linear C 6-9 alkylene, linear C 6-9 alkenylene and linear C 6-9 alkynylene
  • R 3 is independently selected from the group consisting of linear C 6-9 alkyl, linear C 6-9 alkenyl and linear C 6-9 alkynyl.
  • R 2 is independently a linear C 6-9 alkylene group or a linear C 6-9 alkenylene group
  • R 3 is independently a linear C 6-9 alkyl group or a linear C 6-9 alkenyl group.
  • R 2 and R 3 are not unsaturated hydrocarbon groups at the same time.
  • M 3 and M 4 are each independently a linear C 10-20 alkenyl or a linear C 10-20 alkynyl.
  • M 3 and M 4 are each independently a linear C 14-18 alkenyl or a linear C 14-18 alkynyl.
  • M 3 and M 4 are each independently a linear C 18 alkenyl or a linear C 18 alkynyl.
  • M 3 and M 4 are linear C 18 alkenyl groups.
  • R 1 is independently selected from the group consisting of hydroxy substituted C 1-6 alkyl, hydroxy substituted C 2-6 alkenyl, and hydroxy substituted C 2-6 alkynyl.
  • R 1 is a C 1-6 alkyl group substituted with a hydroxy group.
  • R 1 is selected from the group consisting of -CH 2 OH, -(CH 2 ) 2 OH, -(CH 2 ) 3 OH, -(CH 2 ) 4 OH, -(CH 2 ) 5 OH, and -(CH 2 ) 6 OH.
  • M 1 and M 2 are each independently an optionally substituted C 1-4 alkylene, and when M 1 and M 2 are C 2 alkylene, R 1 is not -(CH 2 ) 6 OH.
  • M 1 and M 2 are each independently an optionally substituted C 3-4 alkylene.
  • n 2, 3, 4 or 5;
  • n is independently 3 or 4;
  • L1 and L2 are each independently selected from the group consisting of -C(O)O-, -OC(O)-, -O-, -C(O)N(H)- and -N(H)C(O)-;
  • M3 and M4 are each independently wherein R 2 is independently selected from the group consisting of linear C 4-10 alkylene, linear C 4-10 alkenylene and linear C 4-10 alkynylene, and R 3 is independently selected from the group consisting of linear C 4-10 alkyl, linear C 4-10 alkenyl and linear C 4-10 alkynyl.
  • R 2 is independently selected from the group consisting of linear C 6-9 alkylene, linear C 6-9 alkenylene and linear C 6-9 alkynylene
  • R 3 is independently selected from the group consisting of linear C 6-9 alkyl, linear C 6-9 alkenyl and linear C 6-9 alkynyl.
  • R 2 is independently a linear C 6-9 alkylene group or a linear C 6-9 alkenylene group
  • R 3 is independently a linear C 6-9 alkyl group or a linear C 6-9 alkenyl group.
  • R 2 and R 3 are not unsaturated hydrocarbon groups at the same time.
  • M 3 and M 4 are each independently a linear C 10-20 alkenyl or a linear C 10-20 alkynyl.
  • M 3 and M 4 are each independently a linear C 14-18 alkenyl or a linear C 14-18 alkynyl.
  • M 3 and M 4 are each independently a linear C 18 alkenyl or a linear C 18 alkynyl.
  • M 3 and M 4 are linear C 18 alkenyl groups.
  • n 2, 3 or 4;
  • L1 and L2 are each independently selected from the group consisting of -C(O)O-, -OC(O)-, -CO-, -C(O)N(H)- and -N(H)C(O)-;
  • M3 and M4 are each independently wherein R 2 is independently a linear C 4-10 alkylene group or a linear C 4-10 alkenylene group, and R 3 is independently a linear C 6-9 alkyl group or a linear C 4-10 alkenyl group.
  • R 2 is independently a linear C 6-9 alkylene group or a linear C 6-9 alkenylene group
  • R 3 is independently a linear C 6-9 alkyl group or a linear C 6-9 alkenyl group.
  • R 2 and R 3 are not unsaturated hydrocarbon groups at the same time.
  • R 2 and R 3 are each independently a linear C 6-9 alkylene group.
  • M 3 and M 4 are linear C 10-20 alkenyl or linear C 10-20 alkynyl.
  • M 3 and M 4 are linear C 14-18 alkenyl or linear C 14-18 alkynyl.
  • M 3 and M 4 are linear C 18 alkenyl groups.
  • M3 and M4 are each independently selected from:
  • n 2 or 3;
  • L1 and L2 are each independently selected from the group consisting of -C(O)O-, -OC(O)-, -CO-, -C(O)N(H)- and -N(H)C(O)-.
  • L 1 and L 2 are each independently selected from -C(O)O-, -OC(O)-, -O-, -C(O)N(H)-, and -N(H)C(O)-.
  • L 1 and L 2 are -C(O)O-.
  • L 1 and L 2 are -OC(O)-.
  • L 1 and L 2 are -CO-.
  • L 1 and L 2 are -C(O)N(H)-.
  • L 1 and L 2 are -N(H)C(O)-.
  • the present application provides a nanoparticle composition comprising a lipid component of the compound described herein.
  • the lipid component further comprises phospholipids.
  • 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 (DPPC), Phosphocholine (POPC), 1,2-di-O-octadecenyl-sn-glycero-3-phosphocholine (18:0Diether PC), 1-oleoyl-2-
  • the phospholipid is DOPE.
  • the phospholipid is DSPC.
  • the lipid component further comprises a structural lipid.
  • the structured lipid is selected from the group consisting of cholesterol, coprosterol, sitosterol, ergosterol, campesterol, stigmasterol, brassicasterol, tomatidine, ursolic acid, ⁇ -tocopherol, and mixtures thereof.
  • the structured lipid is cholesterol
  • the lipid component further comprises PEG lipids.
  • 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.
  • the PEG lipid comprises a PEG moiety having a size of about 1000 Da to about 20 kDa.
  • 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.
  • the PEG lipid is PEG2000-DMG.
  • the lipid component further comprises phospholipids, structural lipids and PEG lipids.
  • 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.
  • 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.
  • 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.
  • the nanoparticle composition further comprises a therapeutic and/or prophylactic agent.
  • 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.
  • the therapeutic and/or prophylactic agent is a nucleic acid, a protein, a peptide, or a small molecule.
  • the therapeutic and/or prophylactic agent is a nucleic acid.
  • the therapeutic and/or prophylactic agent is ribonucleic acid (RNA).
  • 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.
  • siRNA small interfering RNA
  • aiRNA asymmetric interfering RNA
  • miRNA microRNA
  • dsRNA Dicer-substrate RNA
  • shRNA small hairpin RNA
  • mRNA messenger RNA
  • the RNA is mRNA.
  • 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.
  • the therapeutic and/or prophylactic agent comprises components of a gene editing system.
  • the components of the gene editing system comprise a polynucleotide encoding a nuclease.
  • 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.
  • ZFNs zinc finger nucleases
  • TALENs transcription activator-like effector nucleases
  • Cas9 CRISPR-associated protein 9
  • engineered homing endonucleases engineered homing endonucleases.
  • the nanoparticle composition further comprises a guide RNA that targets the nuclease to a specific site in the target cell genome.
  • the components of the gene editing system include: i) CRISPR-associated protein 9 (Cas9), or mRNA encoding CRISPR-associated protein 9 (Cas9); and
  • sgRNA single guide RNA
  • sgRNA single guide RNA
  • the therapeutic and/or prophylactic agent comprises a small molecule.
  • the therapeutic and/or prophylactic agent comprises a PROTAC (Proteolysis Targeting Chimera) molecule.
  • 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.
  • the wt/wt ratio of the lipid component to the therapeutic and/or prophylactic agent is from about 10:1 to about 60:1.
  • the nanoparticle composition has an N:P ratio of about 2:1 to about 30:1.
  • the nanoparticle composition has an average size of about 70 nm to about 100 nm.
  • the present application provides use of the compound described herein and the nanoparticle composition described herein in preparing a delivery system.
  • the delivery system is a CD36-targeted delivery system.
  • the present application provides a delivery system comprising the compound described herein or the nanoparticle composition described herein.
  • the delivery system is a CD36-targeted delivery system.
  • the present application provides a pharmaceutical composition comprising the nanoparticle composition described herein and a pharmaceutically acceptable carrier.
  • 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.
  • the cells express CD36.
  • the mammalian cell is in a mammal.
  • the mammal is a human.
  • 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.
  • the organ or tissue is an organ or tissue that expresses CD36.
  • 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.
  • FIG1 shows the fluorescence photography of DC2.4 cells after transfection of the liposome nanoparticles LNP001 of the present application
  • FIG2 shows the fluorescence photography of A549 cells after transfection of the liposome nanoparticles LNP001 of the present application
  • FIG3A shows the fluorescence expression of the liposome nanoparticles LNP001 and LNP002 of the present application after intramuscular injection
  • FIG3B shows the fluorescence photography of the liposome nanoparticles LNP001 and LNP002 after intravenous injection of the present application
  • FIG3C shows the fluorescence photography of the liposome nanoparticles LNP001 and LNP002 after subcutaneous injection of the present application
  • FIG4 shows the delivery results of the muscle-targeted gene editing system of the liposome nanoparticle LNP001 of the present application
  • FIG5 shows the PROTAC drug delivery results of the liposome nanoparticle LNP001 of the present application targeting Hela cells.
  • alkyl and alkylene generally refer to a straight-chain or branched saturated hydrocarbon moiety.
  • an alkyl group is a straight-chain saturated hydrocarbon.
  • an "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-chain alkyl groups include isopropyl, sec-butyl, isobutyl, tert-butyl, and isopentyl.
  • C 1-14 alkyl means an optionally substituted straight-chain or branched saturated hydrocarbon group containing 1-14 carbon atoms. Unless otherwise specified, alkyl groups as described herein refer to both unsubstituted and substituted alkyl groups.
  • alkenyl and alkenylene generally refer to a straight or branched hydrocarbon moiety having one or more carbon-carbon double bonds.
  • the alkenyl contains 1, 2, or 3 double bonds and is unsaturated. Unless otherwise indicated, an "alkenyl” contains 2-24 carbon atoms.
  • Alkenyl includes cis and trans isomers. Representative straight and branched alkenyls 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.
  • C 2-14 alkenyl means an optionally substituted straight or branched hydrocarbon comprising 2-14 carbon atoms and at least one carbon-carbon double bond.
  • An alkenyl group can include one, two, three, four, or more carbon-carbon double bonds.
  • a C 18 alkenyl group may include one or more double bonds
  • a C 18 alkenyl group including two double bonds may be a linoleyl group.
  • the alkenyl group described herein refers to both unsubstituted and substituted alkenyl groups.
  • alkynyl and alkynylene generally refer to a straight or branched chain hydrocarbon moiety having one or more carbon-carbon triple bonds.
  • an "alkynyl” group 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.
  • C2-14 alkynyl means an optionally substituted straight or branched chain hydrocarbon group comprising 2-14 carbon atoms and at least one carbon-carbon triple bond.
  • alkynyl group may include one, two, three, four, or more carbon-carbon triple bonds.
  • a C18 alkynyl group may include one or more carbon-carbon triple bonds.
  • alkynyl groups as described herein refer to both unsubstituted and substituted alkynyl groups.
  • the terms "optionally substituted alkyl,”"optionally substituted alkenyl,” and “optionally substituted alkynyl” generally mean that when substituted, at least one hydrogen atom is replaced with a substituent.
  • isotopes refer to atoms with the same atomic number but different mass numbers due to the number of neutrons in the nucleus.
  • isotopes of hydrogen include tritium and deuterium.
  • 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.
  • 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)).
  • LNPs lipid nanoparticles
  • liposomes e.g., lipid vesicles
  • lipid complexes e.g., cationic lipid complexes (lipoplex, LPX)
  • a nanoparticle composition can be a liposome with a lipid bilayer having a diameter of 500 nm or less.
  • lipid component generally refers to a component of a nanoparticle composition that includes one or more lipids.
  • the lipid component can include one or more cationic/ionizable lipids, pegylated lipids, structural lipids, or other lipids, such as phospholipids.
  • 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.
  • 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.
  • 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.
  • PEG lipid generally refers to a lipid that comprises a polyethylene glycol component.
  • 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.
  • 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.
  • 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.
  • nucleic acid molecule includes sense and antisense strands, as well as single-stranded and double-stranded forms.
  • nucleic acid molecules described herein may contain naturally occurring or non-naturally occurring nucleotides.
  • non-naturally occurring nucleotides include modified nucleotide bases having derivatized sugar or phosphate backbone linkages or chemically modified residues.
  • RNA generally refers to ribonucleic acid, which may be naturally occurring or non-naturally occurring.
  • RNA may include modified and/or non-naturally occurring components, such as one or more nucleobases, nucleosides, nucleotides, or linkers.
  • RNA may include a cap structure, a chain-terminating nucleoside, a stem-loop, a polyadenylate sequence, and/or a polyadenylation signal.
  • RNA may have a nucleotide sequence encoding a polypeptide of interest.
  • RNA may be messenger RNA (mRNA).
  • RNA may 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.
  • siRNA small interfering RNA
  • aiRNA asymmetric interfering RNA
  • miRNA microRNA
  • dsRNA Dicer-substrate RNA
  • shRNA small hairpin RNA
  • mRNA single-stranded guide RNA
  • cas9 mRNA single-stranded guide RNA
  • the term "contact” generally refers to establishing a physical connection between two or more entities.
  • 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.
  • 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.
  • the nanoparticle composition can contact more than one mammalian cell.
  • 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.
  • 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.
  • a target tissue of interest e.g., MC3, KC2, or DLinDMA
  • 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).
  • 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.
  • certain compounds disclosed herein exhibit similar delivery enhancement when used for intravenous or intramuscular delivery of therapeutic and/or prophylactic agents.
  • 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).
  • a target tissue of interest e.g., 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.
  • 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.
  • 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).
  • 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).
  • 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).
  • in vivo refers to events that occur inside an organism (such as an animal, plant or microorganism, or a cell or tissue thereof).
  • 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.
  • 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.
  • 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.
  • 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).
  • 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.
  • 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.
  • target tissues of interest include specific tissues, organs, and systems or groups thereof.
  • 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.
  • non-target tissues can include the liver and spleen.
  • 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.
  • “treating” cancer may refer to inhibiting the survival, growth and/or spread of a tumor.
  • 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.
  • prevention describes 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.
  • 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.
  • an "individual” or “subject” is a human.
  • 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).
  • 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).
  • 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.
  • words such as "a (kind)” and “said” may represent one (kind) or more than one (kind).
  • 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.
  • 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.
  • 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.
  • 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.
  • the present application provides a compound of formula (I):
  • R 1 can be independently selected from the group consisting of hydroxy-substituted alkyl, hydroxy-substituted alkenyl, and hydroxy-substituted alkynyl;
  • M 1 and M 2 can each be independently selected from the group consisting of optionally substituted alkylene, optionally substituted alkenylene and optionally substituted alkynylene;
  • L1 and L2 can each 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: C1-3 alkyl, C2-3 alkenyl and H;
  • M3 and M4 can each independently be wherein R 2 may be independently selected from the group consisting of a linear C 4-10 alkylene group, a linear C 4-10 alkenylene group and a linear C 4-10 alkynylene group, and R 3 may be independently selected from the group consisting of a linear C 4-10 alkyl group, a linear C 4-10 alkenyl group and a linear C 4-10 alkynyl group.
  • R 2 can be independently selected from the group consisting of linear C 6-9 alkylene, linear C 6-9 alkenylene, and linear C 6-9 alkynylene
  • R 3 can be independently selected from the group consisting of linear C 6-9 alkyl, linear C 6-9 alkenyl, and linear C 6-9 alkynyl.
  • 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.
  • R2 and R3 may not both be unsaturated hydrocarbon groups.
  • R2 is a C4-10 alkylene group
  • R3 is a linear C4-10 alkenyl group or a linear C4-10 alkynyl group.
  • R2 is a linear C4-10 alkenyl group or a linear C4-10 alkynyl group
  • R3 is a linear C4-10 alkyl group.
  • R2 and R3 are both C4-10 alkylene groups.
  • M3 and M4 can each independently be a linear C10-20 alkenyl (e.g., C11-19 alkenyl, C12-18 alkenyl, C13-18 alkenyl, C14-18 alkenyl, C15-18 alkenyl, or C16-18 alkenyl) or a linear C10-20 alkynyl (e.g., C11-19 alkynyl, C12-18 alkynyl, C13-18 alkynyl, C14-18 alkynyl, C15-18 alkynyl, or C16-18 alkynyl ).
  • C10-20 alkenyl e.g., C11-19 alkenyl, C12-18 alkenyl, C13-18 alkenyl, C14-18 alkenyl, C15-18 alkynyl, or C16-18 alkynyl
  • M3 and M4 can each independently be a linear C14-18 alkenyl group (e.g., C14 alkenyl, C15 alkenyl, C16 alkenyl, C17 alkenyl, or C18 alkenyl) or a linear C14-18 alkynyl group (e.g., C14 alkynyl, C15 alkynyl, C16 alkynyl, C17 alkynyl, or C18 alkynyl).
  • C14 alkenyl group e.g., C14 alkenyl, C15 alkenyl, C16 alkenyl, C17 alkenyl, or C18 alkenyl
  • a linear C14-18 alkynyl group e.g., C14 alkynyl, C15 alkynyl, C16 alkynyl, C17 alkynyl, or C18 alkynyl
  • M3 and M4 can each independently be a linear C14-18 alkenyl or a linear C14-18 alkynyl
  • M3 and M4 can each independently be wherein R 2 may be independently selected from a linear C 4-10 alkylene group (e.g., C 4 alkylene, C 5 alkylene, C 6 alkylene, C 7 alkylene, C 8 alkylene, C 9 alkylene or C 10 alkylene)
  • R 3 may be independently selected from a linear C 4-10 alkyl group (e.g., C 4 alkyl, C 5 alkyl, C 6 alkyl, C 7 alkyl, C 8 alkyl, C 9 alkyl or C 10 alkyl).
  • M 3 and M 4 are each independently a linear C 18 alkenyl group or a linear C 18 alkynyl group.
  • M 3 and M 4 may be linear C 18 alkenyl groups.
  • M3 and M4 are linear C18 alkenyl groups, and M3 and M4 are each independently wherein R 2 is independently selected from a linear C 4-10 alkylene group (e.g., C 4 alkylene, C 5 alkylene, C 6 alkylene, C 7 alkylene, C 8 alkylene, C 9 alkylene or C 10 alkylene) or a linear C 4-10 alkenylene group (e.g., C 4 alkenylene, C 5 alkenylene, C 6 alkenylene, C 7 alkenylene, C 8 alkenylene, C 9 alkenylene or C 10 alkenylene), and R 3 is independently selected from a linear C 4-10 alkyl group (e.g., C 4 alkyl, C 5 alkyl, C 6 alkyl, C 7 alkyl, C 8 alkyl, C 9 alkyl or C 10 alkyl) or a linear C 4-10 alkenyl group (e.g., C 4 alkenyl, C 5 alkenyl,
  • M3 and M4 are linear C18 alkenyl groups, and M3 and M4 are each independently Wherein R 2 is independently selected from a linear C 6 alkylene group, and R 3 is independently selected from a linear C 10 alkyl group.
  • M 3 and M 4 are linear C 18 alkenyl groups, and M 3 and M 4 are each independently Wherein R 2 is independently selected from a linear C 7 alkylene group, and R 3 is independently selected from a linear C 9 alkyl group.
  • M 3 and M 4 are linear C 18 alkenyl groups, and M 3 and M 4 are each independently Wherein R 2 is independently selected from a linear C 8 alkylene group, and R 3 is independently selected from a linear C 8 alkyl group.
  • M 3 and M 4 are linear C 18 alkenyl groups, and M 3 and M 4 are each independently Wherein R 2 is independently selected from a linear C 10 alkylene group, and R 3 is independently selected from a linear C 6 alkyl group.
  • M 3 and M 4 are linear C 18 alkenyl groups, and M 3 and M 4 are each independently Wherein R 2 is independently selected from a linear C 9 alkylene group, and R 3 is independently selected from a linear C 7 alkyl group.
  • M 3 and M 4 are linear C 18 alkenyl groups, and M 3 and M 4 are each independently wherein R 2 is independently selected from a linear C 8 alkylene group, and R 3 is independently selected from a linear C 8 alkenyl group.
  • R1 can be independently selected from the group consisting of hydroxy-substituted C1-6 alkyl, hydroxy-substituted C2-6 alkenyl, and hydroxy-substituted C2-6 alkynyl.
  • R1 is a hydroxy-substituted C1-6 alkyl.
  • R1 can be selected from the group consisting of -CH2OH , -( CH2 ) 2OH , -( CH2 ) 3OH , -( CH2 ) 4OH , -( CH2 ) 5OH , and -( CH2 ) 6OH .
  • R 1 is selected from the group consisting of -CH 2 OH, -(CH 2 ) 2 OH, -(CH 2 ) 3 OH, -(CH 2 ) 4 OH, -(CH 2 ) 5 OH, and -(CH 2 ) 6 OH; and M 3 and M 4 are each independently wherein R 2 is independently selected from the group consisting of linear C 4-10 alkylene, linear C 4-10 alkenylene and linear C 4-10 alkynylene, and R 3 is independently selected from the group consisting of linear C 4-10 alkyl, linear C 4-10 alkenyl and linear C 4-10 alkynyl.
  • M 1 and M 2 are each independently an optionally substituted C 1-4 alkylene, and when M 1 and M 2 are C 2 alkylene, R 1 is not -(CH 2 ) 6 OH.
  • M 1 and M 2 are each independently an optionally substituted C 3-4 alkylene.
  • R 1 is selected from the group consisting of -CH 2 OH, -(CH 2 ) 2 OH, -(CH 2 ) 3 OH, -(CH 2 ) 4 OH, -(CH 2 ) 5 OH, and -(CH 2 ) 6 OH;
  • M 1 and M 2 are each independently an optionally substituted C 3-4 alkylene.
  • R 1 is selected from the group consisting of -CH 2 OH, -(CH 2 ) 2 OH, -(CH 2 ) 3 OH, -(CH 2 ) 4 OH, -(CH 2 ) 5 OH, and -(CH 2 ) 6 OH;
  • M 1 and M 2 are each independently an optionally substituted C 3-4 alkylene;
  • M 3 and M 4 are each independently wherein R 2 is independently selected from the group consisting of linear C 4-10 alkylene, linear C 4-10 alkenylene and linear C 4-10 alkynylene, and R 3 is independently selected from the group consisting of linear C 4-10 alkyl, linear C 4-10 alkenyl and linear C 4-10 alkynyl.
  • R 1 is selected from the group consisting of -CH 2 OH, -(CH 2 ) 2 OH, -(CH 2 ) 3 OH, -(CH 2 ) 4 OH, -(CH 2 ) 5 OH, and -(CH 2 ) 6 OH;
  • M 1 and M 2 are each independently an optionally substituted C 3-4 alkylene group;
  • M 3 and M 4 are each independently a linear C 14-18 alkenyl group or a linear C 14-18 alkynyl group, and M 3 and M 4 are each independently wherein R 2 is independently selected from the group consisting of linear C 4-10 alkylene, linear C 4-10 alkenylene and linear C 4-10 alkynylene, and R 3 is independently selected from the group consisting of linear C 4-10 alkyl, linear C 4-10 alkenyl and linear C 4-10 alkynyl.
  • R 1 is selected from the group consisting of -CH 2 OH, -(CH 2 ) 2 OH, -(CH 2 ) 3 OH, -(CH 2 ) 4 OH, -(CH 2 ) 5 OH, and -(CH 2 ) 6 OH;
  • M 1 and M 2 are each independently an optionally substituted C 3-4 alkylene;
  • M 3 and M 4 are linear C 18 alkenyl, and M 3 and M 4 are each independently wherein R 2 is independently a linear C 6-9 alkylene group or a linear C 6-9 alkenylene group, and R 3 is independently a linear C 6-9 alkyl group or a linear C 6-9 alkenyl group.
  • R 1 is selected from the group consisting of -CH 2 OH, -(CH 2 ) 2 OH, -(CH 2 ) 3 OH, -(CH 2 ) 4 OH, -(CH 2 ) 5 OH, and -(CH 2 ) 6 OH;
  • M 1 and M 2 are each independently an optionally substituted C 3-4 alkylene; wherein M 3 and M 4 are each independently selected from:
  • n 2, 3, 4 or 5;
  • n is independently 3 or 4;
  • L1 and L2 are each independently selected from the group consisting of -C(O)O-, -OC(O)-, -O-, -C(O)N(H)- and -N(H)C(O)-;
  • M3 and M4 are each independently wherein R 2 is independently selected from the group consisting of linear C 4-10 alkylene, linear C 4-10 alkenylene and linear C 4-10 alkynylene, and R 3 is independently selected from the group consisting of linear C 4-10 alkyl, linear C 4-10 alkenyl and linear C 4-10 alkynyl.
  • R 2 is independently selected from the group consisting of linear C 6-9 alkylene, linear C 6-9 alkenylene and linear C 6-9 alkynylene
  • R 3 is independently selected from the group consisting of linear C 6-9 alkyl, linear C 6-9 alkenyl and linear C 6-9 alkynyl.
  • R 2 is independently a linear C 6-9 alkylene group or a linear C 6-9 alkenylene group
  • R 3 is independently a linear C 6-9 alkyl group or a linear C 6-9 alkenyl group.
  • R2 and R3 are not both unsaturated hydrocarbon groups.
  • R2 is a C6-9 alkylene group
  • R3 is a linear C6-9 alkenyl group.
  • R2 is a linear C6-9 alkenyl group
  • R3 is a linear C6-9 alkyl group.
  • R2 and R3 are both C6-9 alkylene groups.
  • M3 and M4 are each independently a linear C10-20 alkenyl or a linear C10-20 alkynyl.
  • M3 and M4 are each independently a linear C14-18 alkenyl or a linear C14-18 alkynyl.
  • M3 and M4 are each independently a linear C18 alkenyl or a linear C18 alkynyl.
  • M 3 and M 4 are linear C 18 alkenyl.
  • M 3 and M 4 are linear C 18 alkenyl
  • M 3 and M 4 are each independently wherein R 2 is independently a linear C 6-9 alkylene group or a linear C 6-9 alkenylene group
  • R 3 is independently a linear C 6-9 alkyl group or a linear C 6-9 alkenyl group.
  • M 3 and M 4 are linear C 18 alkenyl groups
  • M 3 and M 4 are each independently wherein R 2 is independently a linear C 6-9 alkylene group, and R 3 is independently a linear C 6-9 alkyl group.
  • n 2, 3 or 4;
  • L1 and L2 are each independently selected from the group consisting of -C(O)O-, -OC(O)-, -CO-, -C(O)N(H)- and -N(H)C(O)-;
  • M3 and M4 are each independently wherein R 2 is independently a linear C 4-10 alkylene group or a linear C 4-10 alkenylene group, and R 3 is independently a linear C 6-9 alkyl group or a linear C 4-10 alkenyl group.
  • R 2 is independently a linear C 6-9 alkylene group or a linear C 6-9 alkenylene group
  • R 3 is independently a linear C 6-9 alkyl group or a linear C 6-9 alkenyl group.
  • R 2 and R 3 are not both unsaturated hydrocarbon groups.
  • R 2 and R 3 are each independently a linear C 6-9 alkylene group.
  • m is 2, 3 or 4;
  • L1 and L2 are each independently selected from the group consisting of -C(O)O-, -OC(O)-, -CO-, -C(O)N(H)- and -N(H)C(O)-;
  • M3 and M4 are each independently wherein R 2 and R 3 are each independently a linear C 6-9 alkylene group.
  • M3 and M4 are linear C10-20 alkenyl or linear C10-20 alkynyl.
  • M3 and M4 are linear C14-18 alkenyl or linear C14-18 alkynyl.
  • M 3 and M 4 are linear C 18 alkenyl. In some embodiments, wherein M 3 and M 4 are linear C 18 alkenyl.
  • M 3 and M 4 are linear C 18 alkenyl, and M 3 and M 4 are each independently wherein R 2 is independently a linear C 6-9 alkylene group or a linear C 6-9 alkenylene group, and R 3 is independently a linear C 6-9 alkyl group or a linear C 6-9 alkenyl group.
  • M 3 and M 4 are linear C 18 alkenyl groups, and M 3 and M 4 are each independently wherein R 2 is independently a linear C 6-9 alkylene group, and R 3 is independently a linear C 6-9 alkyl group.
  • M3 and M4 are each independently selected from:
  • m is 2, 3 or 4;
  • L1 and L2 are each independently selected from the group consisting of -C(O)O-, -OC(O)-, -CO-, -C(O)N(H)- and -N(H)C(O)-;
  • M3 and M4 are
  • m is 2, 3 or 4;
  • L1 and L2 are each independently selected from the group consisting of -C(O)O-, -OC(O)-, -CO-, -C(O)N(H)- and -N(H)C(O)-;
  • M3 and M4 are
  • n 2 or 3;
  • L1 and L2 are each independently selected from the group consisting of -C(O)O-, -OC(O)-, -CO-, -C(O)N(H)- and -N(H)C(O)-.
  • L1 and L2 are each independently selected from -C(O)O-, -OC(O)-, -O-, -C(O)N(H)-, and -N(H)C(O)-.
  • L1 and L2 are -C(O)O-.
  • L1 and L2 are -OC(O)-.
  • L1 and L2 are -CO-.
  • L1 and L2 are -C(O)N(H)-.
  • L1 and L2 are -N(H)C(O)-.
  • 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.
  • 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-diondecanoyl-sn-glycero-phosphocholine (DUPC), 1-palmitoyl-2-oleoyl-sn- Glycerol-3-phosphocholine (POPC), 1,2-di-O-octadecenyl-sn-glycerol
  • the lipid component of the nanoparticle composition can include a structured lipid.
  • the structured lipid can be selected from the group consisting of cholesterol, coprosterol, sitosterol, ergosterol, campesterol, stigmasterol, brassicasterol, tomatine, ursolic acid, ⁇ -tocopherol, and mixtures thereof.
  • the structured lipid can be cholesterol.
  • 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.
  • 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).
  • PEG-DMG 1,2-dimyristoyl-sn-glyceromethoxypolyethylene glycol
  • PEG-DSPE 1,2-distearoyl-sn-g
  • the PEG lipid comprises a PEG moiety having a size of about 1000 Da to about 20 kDa.
  • 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.
  • the PEG lipid is PEG2000-DMG.
  • the lipid component further comprises phospholipids, structural lipids and PEG lipids.
  • 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.
  • a phospholipid such as an unsaturated lipid, such as DOPE or DSPC
  • a PEG lipid such as an unsaturated lipid, such as DOPE or DSPC
  • the ingredients of the provided lipid component can be in a specific ratio.
  • 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.
  • 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.
  • 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.
  • the phospholipid can be DOPE or DSPC.
  • the PEG lipid can be PEG-DMG and/or the structural lipid can be cholesterol.
  • 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.
  • 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.
  • 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.
  • 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.
  • the therapeutic and/or prophylactic agent is a nucleic acid, a protein, a peptide, or a small molecule.
  • the therapeutic and/or preventive agent is a nucleic acid.
  • 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.
  • the therapeutic and/or preventive agent is RNA.
  • 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.
  • siRNA small interfering RNA
  • aiRNA asymmetric interfering RNA
  • miRNA microRNA
  • dsRNA Dicer-substrate RNA
  • shRNA small hairpin RNA
  • tRNA transfer RNA
  • mRNA messenger RNA
  • 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.
  • the polypeptide encoded by the mRNA can have a therapeutic effect when expressed in a cell.
  • 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.
  • 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.
  • 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.
  • siRNA can be immunomodulatory siRNA.
  • 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.
  • the therapeutic agent and/or preventive agent comprises a component of a gene editing system.
  • gene editing and its grammatical equivalents may refer to a genetic engineering that inserts, replaces or removes one or more nucleotides from a genome.
  • 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.
  • Cas protein such as Cas9
  • ZFN zinc finger nuclease
  • TALEN transcription activator-like effector nuclease
  • the nuclease may be a naturally occurring nuclease, a genetically modified nuclease and/or a recombinant nuclease.
  • 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.
  • 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).
  • ZFNs zinc finger nucleases
  • TALENs transcription activator-like effector nucleases
  • Cas9 CRISPR-associated protein 9
  • engineered homing endonucleases engineered homing endonucleases
  • the nanoparticle composition further comprises a guide RNA that targets the nuclease to a specific site in the genome of the target cell.
  • the guide RNA can comprise two RNA molecules, referred to herein as “dual guide RNA” or “dgRNA.”
  • the guide RNA can comprise a single RNA molecule, referred to herein as “single guide RNA” or “sgRNA.”
  • 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).
  • the therapeutic and/or prophylactic agent can be an sgRNA and/or cas9 mRNA.
  • the sgRNA and/or cas9 mRNA can be used as a gene editing tool.
  • the sgRNA-cas9 complex can affect the mRNA translation of a cellular gene.
  • 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.
  • EPO erythropoietin
  • G-CSF granulocyte colony stimulating factor
  • GM-CSF granulocyte-macrophage colony stimulating factor
  • LHRH luteinizing hormone-releasing hormone
  • the therapeutic and/or prophylactic agent comprises a small molecule.
  • 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.
  • 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.
  • PROTAC therefore works by allowing the ligand/molecule to bind to the ubiquitin ligase, thereby recruiting the target protein of interest to the ligase for ubiquitination and ultimately proteolysis and degradation.
  • 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.
  • 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.
  • 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.
  • 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.
  • the wt/wt ratio of the lipid component to the therapeutic and/or prophylactic agent can be about 10: 1 to about 40: 1.
  • 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).
  • 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.
  • 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.
  • Nanoparticle compositions can be characterized by a variety of methods. For example, the morphology and size distribution of a nanoparticle composition can be examined using microscopy (e.g., transmission electron microscopy or scanning electron microscopy). Dynamic light scattering or potentiometric analysis (e.g., potentiometric titration) can be used to measure the zeta potential. Dynamic light scattering can also be used to determine particle size. Instruments such as the Zetasizer Nano ZS (Malvern Instruments Ltd, Malvern, Worcestershire, UK) can also be used to measure various characteristics of a nanoparticle composition, such as particle size, polydispersity index, and zeta potential.
  • microscopy e.g., transmission electron microscopy or scanning electron microscopy
  • Dynamic light scattering or potentiometric analysis e.g., potentiometric titration
  • Instruments such as the Zetasizer Nano ZS (Malvern Instruments Ltd, Malver
  • 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).
  • 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.
  • 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, from about
  • 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.
  • 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.
  • the zeta potential of a nanoparticle composition can be used to indicate the zeta potential of the composition.
  • 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.
  • 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 0 mV to about +15 mV, about
  • Nanoparticle compositions can be formulated as whole or as part of a pharmaceutical composition.
  • a pharmaceutical composition can include one or more nanoparticle compositions.
  • 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.
  • 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.
  • 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.
  • 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.
  • compositions can be prepared into various forms suitable for a variety of routes and methods of administration.
  • 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.
  • 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.
  • FAT class B scavenger receptor family
  • SCARB3 glycoprotein IV
  • GP88 glycoprotein IV
  • gpIV glycoprotein IV
  • gpIIIb glycoprotein IIIb
  • 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 ⁇ -amyloid (fA ⁇ ); dying/apoptotic cells; glycated proteins; and phospholipids.
  • oxLDL, mLDL long-chain fatty acids
  • lipid and lipoprotein components of bacterial cell walls lipid and lipoprotein components of bacterial cell walls
  • TSP thrombospondins
  • TSR type 1 thrombospondin repeat
  • 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.
  • phagocytes such as macrophages
  • ligand such as lipids and fatty acids
  • 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.
  • therapeutic and/or prophylactic agents e.g., RNA
  • Nanoparticle compositions can be designed for one or more specific applications or targets.
  • 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).
  • therapeutic and/or prophylactic agents such as RNA
  • 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.
  • protein, cytotoxic agent, radioactive ion, chemotherapeutic agent or nucleic acid can utilize nanoparticle composition to be delivered to cell or organ.
  • RNA for example mRNA
  • translatable mRNA can produce paid close attention to polypeptide in cell translation.
  • 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).
  • 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.
  • the destination of interest e.g., tissue expressing CD36
  • tissue of interest e.g., tissue of interest, such as tissue expressing CD36
  • 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.
  • 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).
  • the present application provides uses of the compounds and nanoparticle compositions described herein in preparing delivery systems.
  • the present application provides a delivery system comprising the compound described herein or the nanoparticle composition described herein.
  • the delivery system is a CD36-targeted delivery system.
  • 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.
  • 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.
  • the cells, tissues and/or organs express CD36.
  • the cells, tissues and/or organs highly express CD36.
  • the mammalian cell is in vivo or in vitro in a mammal.
  • the mammal is a human.
  • 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.
  • the present application also provides use of the nanoparticle composition described herein in preparing medicines.
  • 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.
  • 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.
  • 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.
  • Nanoparticle compositions containing one or more therapeutic and/or prophylactic agents can be administered by any route.
  • 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
  • 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.
  • the present disclosure encompasses delivery or administration of the compositions described herein by any appropriate route.
  • 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.
  • 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
  • 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.
  • 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.
  • each agent will be administered at a dose and/or time course determined for that agent.
  • 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.
  • 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.
  • Step 2 Synthesis of di((Z)-octadec-9-en-1-yl)4,4'-((2-hydroxyethyl)azanediyl)dibutyrate
  • 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.
  • Step 2 Synthesis of 4,4'-((2-hydroxyethyl)azanediyl)bis(N-((Z)-octadec-9-en-1-yl)butanamide
  • Step 2 Synthesis of 2-(bis(4-(((Z)-octadec-9-en-1-yl)oxy)butyl)amino)ethan-1-ol
  • Step 2 Synthesis of di((Z)-octadec-9-en-1-yl)4,4'-((3-hydroxypropyl)azanediyl)dibutyrate
  • Step 2 Synthesis of 4,4'-((3-hydroxypropyl)azanediyl)bis(N-((Z)-octadec-9-en-1-yl)butanamide)
  • Step 2 Synthesis of 3-(bis(4-(((Z)-octadec-9-en-1-yl)oxy)butyl)amino)propan-1-ol
  • the first step is the synthesis of di-tert-butyl(((2-hydroxyethyl)azanediyl)bis(propane-3,1-diyl))dicarbamate
  • 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 spin drying.
  • Step 3 Synthesis of (10Z,10'Z)-N,N'-(((2-hydroxyethyl)azanediyl)bis(propane-3,1-diyl))bis(nonadec-10-enamide)
  • Step 1 Synthesis of di-tert-butyl(((3-hydroxypropyl)azanediyl)bis(propane-3,1-diyl))dicarbamate
  • Step 3 Synthesis of (10Z,10'Z)-N,N'-(((3-hydroxypropyl)azanediyl)bis(propane-3,1-diyl))bis(nonadec-10-enamide)
  • Step 2 Synthesis of ((3-hydroxypropyl)azanediyl)bis(propane-3,1-diyl)(10Z,10'Z)-bis(nonadec-10-enoate)
  • 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.
  • Example 12 Transfection of DC2.4 cells with eGFP mRNA using liposome nanoparticles (LNPs)
  • DC2.4 is a dendritic cell lineage that has been shown to express CD36.
  • DMEM Dulbecco's Modified Eagle's Medium
  • the cells Prior to transfection, the cells were aspirated, washed once with DPBS, and then incubated in serum-reduced Opti-MEM medium 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. The next day, the 24-well plate was directly photographed for fluorescence, digested normally, and cells were counted using trypan blue staining, and fluorescence values were measured by flow cytometry.
  • 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%.
  • Example 13 Transfection of A549 cells with eGFP mRNA using the liposome nanoparticles (LNPs):
  • 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 of 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.
  • LNPs containing 200 ng of eGFP-mRNA were added to each well and incubated overnight at 37°C. The next day, the 24-well plate was directly photographed for fluorescence and digested as usual. Cells were counted using trypan blue staining and fluorescence was measured by flow cytometry.
  • 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%.
  • Group Reference 1 After grouping, drug administration began on the first day.
  • Group Reference 2 Group Reference 2
  • Group A, Group B, Group C, Group D and Group E were administered by tail vein injection/muscular injection/subcutaneous injection, respectively. All were single doses, and the drug was administered once in total.
  • mice in Group 1 and Group E Six hours after the start of drug administration, the whole-body fluorescence imaging of mice in Group 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 1 was 7.62E+05p/s; the bioluminescence values of the heart, liver, spleen, lung, kidney and lymph nodes of Group 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, 4.55
  • 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.
  • 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 (the fluorescence values reported for the intramuscular injection site and liver in Reference 2 and Group A/B can be compared). Because bioluminescence values for subcutaneous 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 ⁇
  • the total bioluminescence value of the tail vein administration group in group E was 1.21E+07 ⁇ 6.03E+06 p/s; the total bioluminescence value of the intramuscular administration group in group E was 1.79E+07 ⁇ 8.22E+06 p/s.
  • 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 extracted and reverse transcribed into cDNA using a Qiagen kit.
  • the gene editing efficiency of the model CRISPR system delivered by different lipid nanoparticles was evaluated 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-targeted nanocomplex group, the targeted LNP001 group significantly improved the gene editing efficiency in the target tissue.
  • Example 16 Targeted drug delivery of anti-tumor PROTAC using the lipid nanoparticles
  • ARV-771 (purchased from MedChem Express) is a PROTAC molecule targeting BRD4.
  • ARV-771 was dissolved in DMSO and then in PBS to a 100 nM solution. Lipo2000 or LNP001 was added to ARV-771 to a final concentration of 5 ⁇ g/ml and incubated for 15 minutes. Then, Lipo2000-AR-771 or LPN001-ARV-771 was added to the pre-prepared six-well plates of 1 ⁇ 10 6 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 targeting nanocomplex group Lipo2000-AR-771, LPN001-ARV-771 carrying the PROTAC molecule targeting BRD4 significantly improved the degradation efficiency of Hela cell BRD4 induced by AR-771.

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Abstract

La présente demande concerne de nouveaux composés lipidiques cationiques et une composition contenant les composés. La présente demande concerne également une composition de nanoparticules qui contient de nouveaux composés lipidiques cationiques et d'autres lipides, tels que des phospholipides, des lipides structuraux et des lipides PEG. De plus, la composition de nanoparticules selon la présente demande peut être utilisée pour introduire un agent thérapeutique et/ou un agent prophylactique dans des cellules ou des organes de mammifère pour, par exemple, réguler l'expression d'un polypeptide, d'une protéine ou d'un gène.
PCT/CN2025/082780 2024-03-20 2025-03-14 Composés lipidiques cationiques, composition et utilisation associées Pending WO2025195308A1 (fr)

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